A-i apolipoprotein mimetics

FIELD: biotechnologies.

SUBSTANCE: peptide is characterised with sequence Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO: 16). The invention also relates to peptide/lipid complex based on the above peptide, in which phospholipid represents one or more of sphingomyelin, DPPC and DPPG, a pharmaceutical composition that contains it, and treatment methods of dyslipidemia, cardiovascular disease, endothelial malfunction, macrovascular illnesses and microvascular illnesses using it.

EFFECT: invention allows obtaining ApoA-I mimetic that is more stable in comparison to ApoA-I and that is easy to obtain.

14 cl, 21 dwg, 14 tbl, 24 ex

 

Cross-reference to related applications

In this application claimed priority according to the date of priority of provisional applications U.S. serial No. 61/152962, filed February 16, 2009, 61/152966, filed February 16, 2009, and 61/152960, filed February 16, 2009, the complete contents of which are incorporated into this description by reference.

The scope of the invention

The present invention relates to peptides, compositions containing them and methods of treatment or prevention of dyslipidemia, cardiovascular disease, endothelial dysfunction, macrovascular disorder or microvascular disorders.

Prior art

Cholesterol circulating in the body, is carried by lipoproteins in the plasma, which are particles of a complex of lipid and protein composition, which carry out the transport of lipids in the blood. Two types of plasma lipoproteins, which carry cholesterol are low-density lipoprotein ("LDL") and high-density lipoprotein ("HDL"). It is believed that LDL is responsible for the delivery of cholesterol from the liver (where it is synthesized or where delivered from food sources) to the body tissues outside of the liver. On the other hand, it is believed that HDL facilitate the transport of cholesterol from extrahepatic t is Anya in the liver, where cholesterol catabolized and subjected to elimination. Such transport of cholesterol from extrahepatic tissues to the liver is called "reverse transport of cholesterol.

The path in reverse cholesterol transport ("RCT") has three main stages: (i) the output of cholesterol, that is, the initial removal of cholesterol from the different pools of peripheral cells; (ii) esterification of cholesterol by the action of lecithin-cholesterol-acyltransferase ("LCAT"), thus preventing reverse the sign-post of cholesterol in the cell; and (iii) absorption of ester cholesterol-high density lipoprotein and delivery of complex ester of cholesterol from HDL to the liver cells.

The path RCT mediated by HDL particles. Each HDL particle has a lipid component and a protein component. The lipid component of HDL may be a phospholipid, cholesterol (or a complex ester of cholesterol or triglyceride. The protein component of HDL initially consists of ApoA-I ApoA-I is synthesized by the liver and small intestine in the form of pre-Pro-apolipoprotein, which is secreted as a Pro-protein that is rapidly cleaved with getting a Mature polypeptide having the 243 amino acid residue. ApoA-I initially consists of 6-8 different recurring items, consisting of 22 amino acid residues that are spatially separated by the linker is the first fragment, which is often Proline, and in some cases it consists of several residues. ApoA-I forms three types of stable complexes with lipids: small, depleted lipid complexes, the so-called HDL pre-β-1; flattened disc-shaped particles containing polar lipids (phospholipids and cholesterol), the so-called HDL pre-β-2; and spherical particles containing both polar and nonpolar lipids, the so-called spherical or Mature HDL (HDL3and HDL2).

Attempts were made to recombinant production, and the introduction of ApoA-I to patients with the aim of protection from atherosclerotic disease. However, with the production and use of ApoA-I associated with many difficulties, which makes it less than ideal as a drug; for example, ApoA-I is a major protein which is difficult and expensive to produce, and you will need to bypass significant difficulties associated with the problems of production and reproduction, in regard to stability during storage, delivery of the active product and the half-life ofin vivo.

Given these disadvantages, attempts have been made to obtain peptides that mimic the activity of ApoA-Iin vivo. In this area there is a need to develop additional peptides, which could mimic the activity of ApoA-Iin vivothe products which shall be simple and justifiable expenditure.

Brief description of the invention

In one of the embodiments of the present invention is associated with peptides containing from 22 to 29 residues having the formula I:

R1-Y1-X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-X18-X19-X20-X21-X22-X23-Y2-R2

The formula I

and their pharmaceutically acceptable salts, in which

X1is missing or achiral, D -, or L-basic amino acid residue;

X2is achiral, D -, or L-aliphatic amino acid residue;

X3is achiral, D -, or L-aliphatic amino acid residue;

X4is achiral, D -, or L-basic amino acid residue;

X5represents Gln, Asn, D-Gln, D-Asn or basically achiral amino acid residue, D - or L-basic amino acid residue;

X6is achiral, D -, or L-basic amino acid residue;

X7is achiral, D -, or L-hydrophobic amino acid residue;

X8is achiral, D -, or L-hydrophobic amino acid residue;

X9is achiral, D -, or L-hydrophobic amino acid residue;

X10represents Leu, Trp, Gly, Nal, D-Leu, D-Trp or D-Nl;

X11represents Gly or achiral, D -, or L-aliphatic amino acid residue;

X12is achiral, D -, or L-hydrophilic amino acid residue;

X13is achiral, D -, or L-hydrophilic amino acid residue;

X14represents Leu, Trp, Gly, D-Leu, or D-Trp;

X15represents Leu, Gly or D-Leu;

X16is achiral, D -, or L-acidic amino acid residue;

X17is achiral, D -, or L-hydrophilic amino acid residue;

X18represents Leu, Phe, D-Leu or D-Phe;

X19represents Leu, Phe, D-Leu or D-Phe;

X20is achiral, D -, or L-acidic amino acid residue;

X21represents Leu, Phe, D-Leu or D-Phe;

X22is achiral, D -, or L-aliphatic amino acid residue; and

X23represents Inp, Nip, azPro, Pip, azPip, D-Nip or D-Pip;

Y1is absent or is an amino acid sequence containing from 1 to 7 residues;

Y2is absent or is an amino acid sequence containing from 1 to 7 residues;

R1represents H or aminosidine group;

R2represents OH or a carboxyl protective group;

a) each chiral amino acid residue is L-amino acid residue;

<> b) each chiral amino acid residue is a D - amino acid residue;

(c) each chiral amino acid residue is L-amino acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is a D-amino acid residue; or

d) each chiral amino acid residue is a D - amino acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is an L-amino acid residue.

In another embodiment of the present invention is associated with peptides, containing from 15 to 22 residues having the following formula II:

R1-Y1-X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-X18-Y2-R2

Formula II

and their pharmaceutically acceptable salts, in which

X1is achiral, D -, or L-basic amino acid residue;

X2represents Leu or D-Leu;

X3is achiral, D -, or L-basic amino acid residue;

X4represents Gln, Asn, D-Gln D-Asn;

X5represents Leu, D-Leu or achiral, D -, or L-basic amino acid residue;

X6represents Leu, Trp, Phe, D-Leu, D-Trp or D-Phe;

X7is achiral, D -, or L-acidic amino acid residue;

X8represents Asn, D-Asn or is achiral, D -, or L-acidic amino acid residue;

X9represents Leu, Trp, D-Leu, or D-Trp;

X10represents Leu, Trp, D-Leu, or D-Trp;

X11is an achiral, D -, or L-acidic amino acid residue;

X12is achiral, D -, or L-basic amino acid residue;

X13represents Leu, Phe, D-Leu or D-Phe;

X14represents Leu, Phe, D-Leu or D-Phe;

X15is achiral, D -, or L-acidic amino acid residue;

X16represents Leu or D-Leu;

X17is achiral, D -, or L-aliphatic amino acid residue;

X18represents Inp, Nip, azPro, Pip, azPip, D-Nip or D-Pip;

Y1is absent or is an amino acid sequence containing from 1 to 4 residues;

Y2no;

R1represents H or aminosidine group;

R2represents OH or a carboxyl protective group;

and from zero to three residues from the X1to X17absent; and

a) each chiral amino is islamnyj residue is L-amino acid residue;

b) each chiral amino acid residue is a D - amino acid residue;

(c) each chiral amino acid residue is L-amino acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is a D-amino acid residue; or

d) each chiral amino acid residue is a D-amino acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is an L-amino acid residue.

In another embodiment of the present invention is associated with peptides containing from 22 to 29 residues having the formula III:

R1-Y1-X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-X18-X19-X20-X21-X22-X23-Y2-R2

Formula III

and their pharmaceutically acceptable salts, in which

X1is missing or achiral, D -, or L-basic amino acid residue;

X2is achiral, D -, or L-basic amino acid residue;

X3the submitted is a Leu or D-Leu;

X4is achiral, D -, or L-basic amino acid residue;

X5is achiral, D -, or L-basic amino acid residue;

X6represents Gln, Asn, D-Gln or D-Asn;

X7represents Leu or D-Leu;

X8represents aAla or D-Ala;

X9represents aAsp or D-Asp;

X10represents aLeu, Phe, Gly, D-Leu or D-Phe;

X11represents Gly, Leu or D-Leu;

X12represents Arg or D-Arg;

X13is achiral, D -, or L-acidic amino acid residue;

X14represents Leu, Trp, Gly, D-Leu, or D-Trp;

X15represents Leu or D-Leu;

X16represents Gln or D-Gln;

X17represents Glu, Leu, D-Glu or D-Leu;

X18represents Leu, Phe, D-Leu or D-Phe;

X19is achiral, D -, or L-aliphatic amino acid residue;

X20represents Glu or D-Glu;

X21represents Leu, Phe, D-Leu or D-Phe;

X22is achiral, D -, or L-aliphatic amino acid residue;

X23represents Inp, Nip, azPro, Pip, azPip, D-Nip or D-Pip;

Y1is absent or is an amino acid sequence containing from 1 to 7 residues;

Y2is absent or is an amino acid sequence containing from 1 on the 7 residues;

R1represents H or aminosidine group;

R2represents OH or a carboxyl protective group;

a) each chiral amino acid residue is L-amino acid residue;

b) each chiral amino acid residue is a D-amino acid residue;

(c) each chiral amino acid residue is L-amino acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is a D-amino acid residue; or

d) each chiral amino acid residue is a D-amino acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is an L-amino acid residue.

The peptide of formula I, II or III, or pharmaceutically acceptable salt ("mimetic ApoA-I) is applicable for treating or preventing dyslipidemia, cardiovascular disease, endothelial dysfunction, macrovascular disorder or microvascular disorders (each being a "Condition").

In another embodiment the present invention relates to compositions containing an effective quantity is the number of mimetica ApoA-I and a pharmaceutically acceptable carrier or excipient.

In another embodiment of the present invention is related to methods of treating or preventing the condition, introducing, if necessary, an effective amount of mimetica ApoA-I to the mammal.

Brief description of the graphical material

Figa is a diagram of Schiffer and Edmundson "helical wheel" (figure "Helical wheel") idealized amphipatic α-helix, in which the hollow circles are hydrophilic amino acid residues, and the filled circles are hydrophobic amino acid residues.

FIGU is a circuit diagram of a spiral grid ("helical net") idealized amphipatic helix depicted in figa.

Figs is a cylindrical chart amphipatic helix depicted in figa.

Figure 2 represents a diagram of Schiffer and Edmundson "helical wheel" 22-dimensional consensus peptide Segrest (SEQ ID NO:1).

On figa illustrated by an extensive network of mimetics of ApoA-I of the third order.

On FIGU illustrated by an extensive network of mimetics of ApoA-I of the fourth order.

On figs illustrated by an extensive network of mimetics of ApoA-I mixed order.

On fig.3D illustrated examples of branched networks "Lys-tree" mimetics of ApoA-I.

Figa is a dynamic from the expression of different States of aggregation and the peptide-lipid complexes which can be obtained using mimetics of ApoA-I according to the invention. Left: the process of multimerization peptides, resulting from the interaction of several peptide helices and leading to the formation of oligomers in conditions with a certain concentration of peptide, pH and ionic strength. In the centre: interaction mimetics of ApoA-I in any of these States of aggregation) with lipid components (e.g., small unilamellar vesicles (SUV,smallunilamellarvesicles)) leads to the reorganization of the lipids. Right: by changing the lipid-peptide molar ratio you can get different types of peptide-lipid complexes, lipid-peptide comical at low lipid-peptide ratios to the disc-shaped particles and, finally, to a large multilamellar complexes with increasing lipid-peptide ratios.

On FIGU illustrated generally accepted model of disc-shaped complexes mimetic ApoA-I-lipid formed at a certain interval ratios mimetic ApoA-I-lipid. Each mimetic ApoA-I, surrounding the rim of the disk is in close contact with his two closest neighbors.

Figure 5 is a representative chromatogram of penetration in gel peptide 16/lipid complex (the lipids are sphingomyelin, DPPC (dipalmitoylphosphatidylcholine and DPPG (dipalmitoylphosphatidylcholine)).

6 represents a curve increase over baseline levels, the fraction of HDL total cholesterol after administration of the peptide 16/lipid complex (the lipids are sphingomyelin, DPPC, and DPPG, and these components are present in a weight ratio of peptide 16:sphingomyelin:DPPC:DPPG as of 1:1.35:1,35:0,30) rabbits.

7 represents a curve of increasing the fraction of HDL free cholesterol after administration of the peptide 16/lipid complex (the lipids are sphingomyelin, DPPC, and DPPG in a weight ratio of 1:1,2125:1,2125:0,075, and the weight ratio of peptide:lipid is 1:2,5) rabbits.

Fig represents the elution profile of gel chromatography on a basic level (dark line) and after 20 min after injection of 2.5 mg/kg of the peptide 16/lipid complex (the lipids are sphingomyelin, DPPC, and DPPG in a weight ratio of 1:1,2125:1,2125:0,075, and the weight ratio of peptide:lipid is 1:2,5) rabbits.

Fig.9 represents a curve of increasing the level of phospholipid in the plasma after administration of the peptide 16/lipid complex (the lipids are sphingomyelin, DPPC, and DPPG in a weight ratio of 1:1,2125:1,2125:0,075, and the weight ratio of peptide:lipid is 1:2,5) rabbits on an empty stomach at doses of 0 (square), 10 (triangles), 20 (circles) or 30 (diamonds) mg/kg Through the different periods of the belts after dose measured the levels of phospholipid in the plasma. The original values (ranging from 0.96 to 1.18 g/l for the four groups) subtracted in order to determine the degree of increased levels of phospholipid in the plasma. Took 3 animals per group. To 30-34-hour period after a dose of these levels returned to values equal to or smaller than the initial level.

Figa represents a curve of increase of total cholesterol in plasma after infusion of a peptide 16/lipid complex (the lipids are sphingomyelin, DPPC, and DPPG in a weight ratio of 1:1,2125:1,2125:0,075, and the weight ratio of peptide:lipid is 1:2,5) rabbits on an empty stomach at doses of 0 (square), 10 (triangles), 20 (circles) or 30 (diamonds) mg/kg Over various time intervals after administration of the dose measured the levels of total cholesterol in the plasma. The original values read in order to determine the degree of increased levels of cholesterol in the plasma. The original values varied from to 0,59 0,77 g/L. Took 3 animals per group. To 30-34-hour period after a dose of these levels returned to values lower than the original level, or equal.

Figv represents a curve of increasing the level of free cholesterol in plasma after infusion of a peptide 16/lipid complex (the lipids are sphingomyelin, DPPC, and DPPG in a weight ratio of 1:,2125:1,2125:0,075, and the weight ratio of peptide:lipid is 1:2,5) rabbits on an empty stomach at doses of 0 (square), 10 (triangles), 20 (circles) or 30 (diamonds) mg/kg Over various time intervals after administration of the dose measured the levels of free cholesterol in the plasma. The original values read in order to determine the degree of increased levels of cholesterol in the plasma. The original values ranged from of 0.21 to 0.27 g/L. Took 3 animals per group. To 30-34-hour period after a dose of these levels returned to values lower than the original level, or equal.

Figs represents a curve of increasing the level of esterified cholesterol in plasma after infusion of a peptide 16/lipid complex (the lipids are sphingomyelin, DPPC, and DPPG in a weight ratio of 1:1,2125:1,2125:0,075, and the weight ratio of peptide:lipid is 1:2,5) rabbits on an empty stomach at doses of 0 (square), 10 (triangles), 20 (circles) or 30 (diamonds) mg/kg Over various time intervals after administration of the dose measured the levels of esterified cholesterol in plasma. The original values read in order to determine the degree of increased levels of cholesterol. The original values ranged from 0.39 to 0.52 g/L. Took 3 animals per group. To 30-34-hour period after a dose of these levels returned to C is achenial, smaller than the original level, or equal.

Figa represents a curve of increasing levels of total cholesterol, HDL cholesterol in plasma after infusion of a peptide 16/lipid complex (the lipids are sphingomyelin, DPPC, and DPPG in a weight ratio of 1:1,2125:1,2125:0,075, and the weight ratio of peptide:lipid is 1:2,5) rabbits on an empty stomach at doses of 0 (square), 10 (triangles), 20 (circles) or 30 (diamonds) mg/kg Over various time intervals after administration of the dose measured the levels of total cholesterol, HDL cholesterol. The original values read in order to determine the degree of increased levels of cholesterol. The initial values of total cholesterol, HDL cholesterol ranged from 0.33 to 0.38 g/L. Took 3 animals per group. To 30-34-hour period after a dose of these levels returned to values lower than the original level, or equal.

Figv represents a curve of increasing the level of free cholesterol, HDL cholesterol in plasma after infusion of a peptide 16/lipid complex (the lipids are sphingomyelin, DPPC, and DPPG in a weight ratio of 1:1,2125:1,2125:0,075, and the weight ratio of peptide:lipid is 1:2,5) rabbits on an empty stomach at doses of 0 (square), 10 (triangles), 20 (circles) or 30 (diamonds) mg/kg Over various time intervals after administration of the dose was measured levels of svobodnolezhaschaya HDL. The original values read in order to determine the degree of increased levels of cholesterol. The original values of free cholesterol, HDL cholesterol ranged from 0.11 to 0.13 g/L. Took 3 animals per group. To 30-34-hour period after a dose of these levels returned to values lower than the original level, or equal.

Figs represents a curve of increasing levels of total cholesterol, HDL cholesterol in plasma after infusion of a peptide 16/lipid complex (the lipids are sphingomyelin, DPPC, and DPPG in a weight ratio of 1:1,2125:1,2125:0,075, and the weight ratio of peptide:lipid is 1:2,5) rabbits on an empty stomach at doses of 0 (square), 10 (triangles), 20 (circles) or 30 (diamonds) mg/kg Over various time intervals after administration of the dose measured the levels of total cholesterol, HDL cholesterol. The original values read in order to determine the degree of increased levels of cholesterol. The initial values of total cholesterol, HDL cholesterol varied from 0.17 to 0.33 g/L. Took 3 animals per group. To 30-34-hour period after a dose of these levels returned to values lower than the original level, or equal.

Fig.11D represents a curve of increasing the level of free cholesterol, HDL cholesterol after infusion of a peptide 16/lipid complex (the lipids are sphingomyelin, DPPC IS DPPG in a weight ratio of 1:1,2125:1,2125:0,075, and the weight ratio of peptide:lipid is 1:2,5) rabbits on an empty stomach at doses of 0 (square), 10 (triangles), 20 (circles) or 30 (diamonds) mg/kg Over various time intervals after administration of the dose measured the levels of free cholesterol in HDL. The original values read in order to determine the degree of increased levels of cholesterol. The original values of free cholesterol, HDL cholesterol ranged from 0.06 to 0.11 g/HP Took 3 animals per group. To 30-34-hour period after a dose of these levels returned to values lower than the original level, or equal.

File represents a curve of increasing levels of total cholesterol, HDL cholesterol after infusion of a peptide 16/lipid complex (the lipids are sphingomyelin, DPPC, and DPPG in a weight ratio of 1:1,2125:1,2125:0,075, and the weight ratio of peptide:lipid is 1:2,5) rabbits on an empty stomach at doses of 0 (square), 10 (triangles), 20 (circles) or 30 (diamonds) mg/kg Over various time intervals after administration of the dose measured the levels of total cholesterol, HDL cholesterol. The original values read in order to determine the degree of increased levels of cholesterol. The original values of free cholesterol, HDL cholesterol ranged from 0.04 to 0.11 g/HP Took 3 animals per group. To 30-34-hour period after a dose of these levels return what was amalis to values smaller than the original level, or equal.

Fig.11F represents a curve of increasing the level of free cholesterol, HDL cholesterol after infusion of a peptide 16/lipid complex (the lipids are sphingomyelin, DPPC, and DPPG in a weight ratio of 1:1,2125:1,2125:0,075, and the weight ratio of peptide:lipid is 1:2,5) rabbits on an empty stomach at doses of 0 (square), 10 (triangles), 20 (circles) or 30 (diamonds) mg/kg Over various time intervals after administration of the dose measured the levels of free cholesterol in HDL. The original values read in order to determine the degree of increased levels of cholesterol. The original values of free cholesterol, HDL cholesterol varied from 0.02 to 0.04 g/L. Took 3 animals per group. To 30-34-hour period after a dose of these levels returned to values lower than the original level, or equal.

Fig represents a curve of increasing the level of triglycerides in plasma after infusion of a peptide 16/lipid complex (the lipids are sphingomyelin, DPPC, and DPPG in a weight ratio of 1:1,2125:1,2125:0,075, and the weight ratio of peptide:lipid is 1:2,5) rabbits on an empty stomach at doses of 0 (square), 10 (triangles), 20 (circles) or 30 (diamonds) mg/kg Over various time intervals after administration of the dose measured the levels of triglycerides in plasma is E. The original values (warioware from 0.40 to 0.80 g/l for the four groups) subtracted in order to determine the degree of increased levels of triglycerides in the plasma. Took 3 animals per group.

Fig represents a curve of increasing levels of free cholesterol, HDL cholesterol in plasma after infusion of a peptide 16/lipid complex (the lipids are sphingomyelin, DPPC, and DPPG in a weight ratio of 1:1,2125:1,2125:0,075, and the weight ratio of peptide:lipid is 1:2,5) rabbits on an empty stomach at doses of 0 (square), and 2.5 (triangles), 5 (circles) or 10 (diamonds) mg/kg At baseline and after 5, 20, 40, 60, 90 and 120 minutes after initiation of infusion was measured levels free cholesterol HDL in plasma. The original values read in order to determine the extent of elevated levels of free cholesterol HDL in plasma. Took 4 animals per group.

Figa represents the elution profile of the gel HPLC-chromatography on a basic level (dark line) and after 20 min after infusion of 2.5 mg/kg of the peptide 16/lipid complex (the lipids are sphingomyelin, DPPC, and DPPG in a weight ratio of 1:1,2125:1,2125:0,075, and the weight ratio of peptide:lipid is 1:2,5). On the Y-axis delayed absorption resulting from continuous computer analysis of free cholesterol of lipoprotein coat is s, eluruumiks with gel column HPLC-chromatography. Peaks from left to right correspond to the fractions of VLDL, LDL and HDL.

Figv represents the elution profile of the gel HPLC-chromatography on a basic level (dark line) and after 20 min after infusion 5.0 mg/kg of the peptide 16/lipid complex (the lipids are sphingomyelin, DPPC, and DPPG in a weight ratio of 1:1,2125:1,2125:0,075, and the weight ratio of peptide:lipid is 1:2,5). On the Y-axis delayed absorption resulting from continuous computer analysis of free cholesterol of lipoprotein fractions, eluruumiks with gel column HPLC-chromatography. Peaks from left to right correspond to the fractions of VLDL, LDL and HDL.

Fig represents a curve of increasing levels of free cholesterol, HDL cholesterol in plasma after infusion of a peptide 16/lipid complex (the lipids are sphingomyelin, DPPC, and DPPG in a weight ratio of 1:1,2125:1,2125:0,075, and the weight ratio of peptide:lipid is 1:2,5) rabbits on an empty stomach in the dose of 20 mg/kg at a rate of 1 ml/min (triangles) or 0.2 ml/min (diamonds). Through various time intervals after administration of the dose measured the levels of free cholesterol, HDL cholesterol in the plasma. The original values read in order to determine the extent of elevated levels of free cholesterol PL is P in the plasma. Took 4 animals per group, the processed peptide 16/lipid complex, and 2 animals per group, the treated filler.

On Fig illustrated by the curves of the kinetic profiles of peptide 16 (top panel), free cholesterol (middle panel) and phospholipid (bottom panel) in male and female rats after administration of the first dose of the peptide 16/lipid complex (the lipids are sphingomyelin, DPPC, and DPPG in a weight ratio of 1:1,2125:1,2125:0,075, and the weight ratio of peptide:lipid is 1:2,5) on day 0. Lower levels of peptide 16 and phospholipid in the plasma over time indicates the clearance of the peptide 16/lipid complex. Presents the kinetics of free cholesterol. Data at each point represent the mean ± SD (N=3 rats/group).

On Fig illustrated by the curves of the kinetic profiles of peptide 16 (top panel), free cholesterol (middle panel) and phospholipid (bottom panel) in male and female rats after administration of multiple doses of the peptide 16/lipid complex (the lipids are sphingomyelin, DPPC, and DPPG in a weight ratio of 1:1,2125:1,2125:0,075, and the weight ratio of peptide:lipid is 1:2,5) on day 26. These animals received the peptide 16/lipid complex every other day for 4 weeks. Lower levels of peptide 16 and phospholipid in the plasma over time indicates the and clearance of the peptide 16/lipid complex. Presents the kinetics of free cholesterol. Data at each point represent the mean ± SD (N=3 rats/group).

On Fig illustrated by the curves of the kinetic profiles of peptide 16 (top panel), free cholesterol (middle panel) and phospholipid (bottom panel) in male and female macaques-Griboedov after the first dose of the peptide 16/lipid complex (the lipids are sphingomyelin, DPPC, and DPPG in a weight ratio of 1:1,2125:1,2125:0,075, and the weight ratio of peptide:lipid is 1:2,5) on day 0. Lower levels of peptide 16 and phospholipid in the plasma over time indicates the clearance of the peptide 16/lipid complex. Presents the kinetics of free cholesterol. Data at each point represent the mean ± SD (N=3 makaka/group).

On Fig illustrated by the curves of the kinetic profiles of peptide 16 (top panel), free cholesterol (middle panel) and phospholipid (bottom panel) in male and female macaques-Griboedov after administration of multiple doses of the peptide 16/lipid complex (the lipids are sphingomyelin, DPPC, and DPPG in a weight ratio of 1:1,2125:1,2125:0,075, and the weight ratio of peptide:lipid is 1:2,5) on day 26. These animals received the peptide 16/lipid complex every other day for 4 weeks. Lower levels of peptide 16 and phospholipid in the plasma from wremen which indicates the clearance of the peptide 16/lipid complex. Presents the kinetics of free cholesterol. Data at each point represent the mean ± SD (N=3 makaka/group).

Figa represents a curve reflecting % increase compared to the level observed before the introduction of dose, total plasma cholesterol in mice C57B1/6J after treatment with composition A, B or C. At different time points the animals were consistently sampled on 6 animals/group.

Figv represents a curve of increasing levels of total cholesterol in plasma in mice C57B1/6J after treatment with composition A, B or C. At different time points the animals were consistently sampled on 6 animals/group.

Figa represents a curve reflecting % increase compared to the level observed before the introduction of dose esterified cholesterol in mice C57B1/6J after treatment with composition A, B or C. At different time points the animals were consistently sampled on 6 animals/group.

Figv represents a curve of increasing levels of esterified cholesterol in mice C57B1/6J after treatment with composition A, B or C. At different time points the animals were consistently sampled on 6 animals/group.

Detailed description of the invention

I.Definitions

The term "approximately", which directly prewar is no numerical or quantitative value, indicates that the specified numeric or quantitative value has the range of variation in average plus or minus 10%. For example, "approximately 1:1" in the range from 0.9:1 to 1.1:1.

Used herein, the term "alkyl", unless otherwise specified, refers to optionally substituted saturated hydrocarbon radical is branched, straight chain or cyclic hydrocarbon radical. Typical alkyl groups are (C1-C6)-alkyl groups, which include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, isopentyl, hexyl, etc. In certain embodiments the alkyl groups are (C1-C4)-alkyl. Unless otherwise specified, the alkyl is unsubstituted.

Used herein, the term "alkenyl", unless otherwise specified, refers to an unsaturated hydrocarbon radical is branched, straight chain or cyclic non-aromatic hydrocarbon radical having one or more carbon-carbon double bonds. These one or more double bond can be either CIS-or TRANS-conformation. Usually alkeneamine groups include, but are not limited to, ethynyl, propenyl, Isopropenyl, butenyl, Isobutanol, tert-butanol, pentanol, hexanol etc. In certain embodiments and chunilna group represents a (C 2-C6)-alkenyl.

Used herein, the term "quinil", unless otherwise specified, refers to an unsaturated hydrocarbon radical is branched or straight chain, having at least one carbon-carbon triple bond. Usually alkyline groups include, but are not limited to, ethinyl, PROPYNYL, butynyl, Isobutanol, pentenyl, hexenyl etc. In certain embodiments Alchemilla group represents a (C2-C6)-quinil.

Used herein, the term "aryl", unless otherwise specified, refers to optionally substituted aromatic system of rings, in which each atom within the ring represents C, O, N or S, thus serving heterocyclic aromatic ring. Usually aryl groups include, but are not limited to, benzyl, phenyl, naphthyl, antracol, furan, imidazole, indazole, indole, isoquinoline, isothiazol, isoxazol, Piran, pyrazin, pyrazole, pyridazine, pyridine, pyrimidine, pyrrole, pyrrolizine, hinzelin, quinoline, hemolysin, cinoxacin, thiazole and thiophene. In certain embodiments of the aryl group represents a (C5-C26-aryl). In certain embodiments the heteroaryl group is a 5-20-membered heteroaryl. Unless otherwise specified, the aryl is unsubstituted.

Used herein, the term "Kalki the", unless otherwise specified, refers to an alkyl group, substituted aryl group.

Used herein, the term "substituted alkyl or aryl", unless otherwise specified, refers to an alkyl or aryl group in which one or more of its hydrogen atoms replaced by another Deputy. Typical substituents include ORa, -SRa, -NRaRa, -NO2, -CN, halogen, -SO2Ra, -C(O)Ra, -C(O)ORaand-C(O)NRaRawhere each Raindependently is hydrogen, alkyl or aryl.

Used herein, the term "hydrophilic shell" unless otherwise specified, refers to the outer surface of the helix, with an overall hydrophilic character.

Used herein, the term "hydrophobic shell" unless otherwise specified, refers to the outer surface of the peptide with an overall hydrophobic character.

Here, when it comes to the mimetic ApoA-I, we mean that the number of end-NH2-groups is equal to zero when R1represents aminosidine group, and is 1 when R1is H.

Here, when it comes to the mimetic ApoA-I, we mean that the number of end-COOH groups is equal to zero when R1represents a carboxyl protective group, and is 1 when R1is OH.

Used herein, the term "mammal"unless otherwise specified refers to a person, mouse, rat, Guinea pig, dog, cat, horse, cow, pig or not belonging to the human race Primate such as a monkey, chimpanzee or baboon. In one of the specified embodiments the mammal is a human.

When the term "effective amount" is used in connection with the mimetic ApoA-I, it means the quantity that is effective for treating or preventing any Condition.

Used herein, the term "free cholesterol HDL" refers to the number of such cholesterol, which has a free hydroxyl group ("free of cholesterol"), which is contained inside the particles of HDL in the serum. HDL particles can be formed from mimetic ApoA-I/lipid complex.

Used herein, the term "total cholesterol, HDL cholesterol" means the amount of cholesterol, which has a hydroxyl group, which etherification ("esterified cholesterol"), which is contained inside the particles of HDL in the serum. HDL particles can be formed from mimetic ApoA-I/lipid complex.

Used herein, the term "amino acid residue", unless otherwise specified, includes a genetically encoded amino acid residues and are not genetically encoded amino acid residues.

Abbreviations for existing used herein, a genetically encoded amino acid residues, n is Evegeny below in table 1.

Table 1
Amino acidSingle-letter abbreviationThe three-letter abbreviation
AlanineAAla
ArginineRArg
AsparagineNAsn
Aspartic acidDAsp
CysteineCCys
GlutamineQGln
Glutamic acidEGlu
GlycineGGly
HistidineHHis
IsoleucineIIle
LLeu
LysineKLys
MethionineMMet
PhenylalanineFPhe
ProlinePPro
SerineSSer
ThreonineTThr
TryptophanWTrp
TyrosineYTyr
ValineVVal

Not genetically encoded amino acid residues include, but are not limited to, β-alanine (β-Ala); 2,3-diaminopropionic acid (Dpr); nicotinebuy acid (Nip); pipecolinic acid (Pip); ornithine (Orn); citrulline (Cit); t-butylene (t-BuA); 2-tert-butylglycol (t-BuG), N-methylisoleucine (MeIle); phenylglycine (PhG); cyclohexylamine (ChA); norleucine (Ne); nafcillin (Nal); 4-chlorophenylalanine (Phe(4-Cl)); 2-forfinally (Phe(2-F)); 3-forfinally (Phe(3-F)); 4-forfinally (Phe(4-F)); penicillamine (Pen); l,2,3,4-tetrahydroisoquinoline-3-carboxylic acid (Tic); β-2-titillans (Thi); methanesulfonic (MSO); homoarginine (hArg); N-acetylized (AcLys); 2,4-diaminobutane acid (Dbu); 2,3-diaminobutane acid (Dab); para-aminophenylalanine (Phe(pNH2)); N-methylvaline (MeVal); homocysteine (hCys), homophenylalanine (hPhe); homoserine (hSer); hydroxyproline (Hyp); gemopolis (hPro); and the corresponding D-enantiomer of each of the above compounds, for example, D-β-Ala, D-Dpr, D-Nip, D-Orn, D-Cit, D-t-BuA, D-t-BuG, D-MeIle, D-PhG, D-ChA, D-Nle, D-Nal, D-Phe(4-Cl), D-Phe(2-F), D-Phe(3-F), D-Phe(4-F), D-Pen, D-Tic, D-Thi, D-MSO, D-hArg, D-AcLys, D-Dbu, D-Dab, D-Phe(pNH2), D is MeVal, D-hCys, D-hPhe, D-hSer, D-Hyp, D-hPro. Other non-genetically encoded amino acid residues include 3-aminopropionic acid; 4-aminobutyric acid; isonicotinic acid (Inp); Aza-pipecolinic acid (azPip); Aza-Proline (azPro); α-aminoadamantane acid (Aib); ε-aminohexanoic acid (Aha); δ-aminovaleric acid (Ava); N-methylglycine (MeGly).

The term "chiral"is used here in relation to amino acid residue, means amino acid residue having at least one chiral center. In one of the embodiments of the specified chiral amino acid residue is L-amino acid residue. Examples of L-amino acid OST tcov include, not limited to, Ala, Arg, Asn, Asp, Cys, Gln, Glu, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, Val, β-Ala, Dpr, Nip, Orn, Cit, t-BuA, t-BuG, MeIle, PhG, ChA, NIe, Nal, Phe(4-Cl), Phe(2-F), Phe(3-F), Phe(4-F), Pen, Tic, Thi, MSO, hArg, AcLys, Dbu, Dab, Phe(pNH2), MeVal, hCys, hPhe, hSer, Hyp and hPro. In one of the embodiments of the specified chiral amino acid residue is a D-amino acid residue. Examples of D-amino acid residues include, but are not limited to, D-Ala, D-Arg, D-Asn, D-Asp, D-Cys, D-Gln, D-Glu, D-His, D-Ile, D-Leu, D-Lys, D-Met, D-Phe, D-Pro, D-Ser, D-Thr, D-Trp, D-Tyr, D-Val, D-β-Ala, D-Dpr, D Nip, D-Pip, D-Orn, D-Cit, D-t-BuA, D-t-BuG, D-MeIle, D-PhG, D-ChA, D-Nle, D-Nal, D-Phe(4-Cl), D-Phe(2-F), D-Phe(3-F), D-Phe(4-F), D-Pen, D-Tic, D-Thi, D-MSO, D-hArg, D-AcLys, D-Dbu, D-Dab, D-Phe(pNH2), D is MeVal, D-hCys, D-hPhe, D-hSer, D-Hyp, D-hPro.

The term "achiral"used here in relation to amino acid residue, means amino acid residue that does not have a chiral center. Examples of achiral amino acid residues include, but are not limited to, Gly, Inp, Aib, Aha, Ava, MeGly, azPip and azPro.

Used herein, the term "aliphatic amino acid residue", unless otherwise specified, refers to an amino acid residue having an aliphatic hydrocarbon chain. Aliphatic amino acid residue includes, but is not limited to, Ala (A)Val (V), Leu (L), Ile (I), Pro (P), azPro, Pip, azPip, β-Ala, Aib, t-BuA, t-BuG, MeIle, ChA, Nle, MeVal, Inp, Nip, hPro, D-Ala, D-Val, D-Leu, D-Ile, D-Pro, D-β-Ala, D-t-BuA, D-t-BuG, D-MeIle, D-Nle, D-MeVal, D-Nip, D-Pip, D-ChA, D-hPro. In one embodied the th specified aliphatic amino acid residue is L-amino acid residue. In another embodiment of the specified aliphatic amino acid residue is a D-amino acid residue. In another embodiment of the specified aliphatic amino acid residue is achiral amino acid residue.

Used herein, the term "hydrophilic amino acid residue", unless otherwise specified, refers to amino acid residue, showing less than zero hydrophobicity, it is generally the normalized hydrophobicity scale of Eisenberg, Eisenberg et al., 1984, J. Mol. Biol. 179:125-142. Hydrophilic amino acid residue includes, but is not limited to, Pro (P), Gly (G), Thr (T)Ser (S), His (H), Glu (E), Asn (N), Gln (Q), Asp (D), Lys (K), Arg (R), Dpr, Orn, Cit, Pen, MSO, hArg, AcLys, Dbu, Dab, Phe(p-NH2), hCys, hSer, Hyp, D-Pro, D-Thr, D-Ser, D-His, D-Glu, D-Asn, D-Gln, D-Asp, D-Lys, D-Arg, D-Dpr, D-Orn, D-Cit, D-Pen, D-MSO, D-hArg, D-AcLys, D-Dbu, D-Dab, D-Phe(p-NH2), D-hCys, D-hSer and D-Hyp. Other hydrophilic amino acid residues include, but are not limited to, analogs of the side chains C1-4with the following formula:

where n is an integer from 1 to 4. In one of the embodiments of the specified hydrophilic amino acid residue is L-amino acid residue. In another embodiment of the specified hydrophilic amino acid residue is a D-amino acid residue. In another embodiment of the specified hydrophilic amino acid is a residue is achiral amino acid residue. In another embodiment of the specified hydrophilic amino acid residue is acidic L-amino acid residue, an acidic D-amino acid residue or an acidic achiral amino acid residue. In another embodiment of the specified hydrophilic amino acid residue is a basic L-amino acid residue, a basic D-amino acid residue or main achiral amino acid residue.

Used herein, the term "hydrophobic amino acid residue", unless otherwise specified, refers to amino acid residue, showing more than zero hydrophobicity, it is generally the normalized hydrophobicity scale of Eisenberg, Eisenberg, 1984, J. Mol. Biol. 179:125-142. Hydrophobic amino acid residue includes, but is not limited to, Ile (I), Phe (F), Val (V), Leu (L), Trp (W), Met (M), Ala (A), Gly (G), Tyr (Y), β-Ala, Nip, t-BuA, t-BuG, MeIle, PhG, ChA, Nle, Nal, Phe(4-Cl), Phe(2-F), Phe(3-F), Phe(4-F), Tic, Thi, MeVal, hPhe, hPro, 3-aminopropionic acid, 4-aminobutyric acid, Inp, Aib, Aha, Ava, MeGly, D-Pro, D-Ile, D-Phe, D-Val, D-Leu, D-Trp, D-Met, D-Ala, D-Tyr, D-β-Ala, D-Nip, D-t-BuA, D-t-BuG, D-MeIle, D-PhG, D-ChA, D-Nle, D-Nal, D-Phe(4-Cl), D-Phe(2-F), D-Phe(3-F), D-Phe(4-F), D-Tic, D-Thi, D is MeVal, D-hPhe and D-hPro. Other hydrophobic amino acids include, but are not limited to, analogs of the side chains C1-4with the following formula:

where n is an integer from 1 to 4. In one of isoprostane specified hydrophobic amino acid residue is L-amino acid residue. In another embodiment of the specified hydrophobic amino acid residue is a D-amino acid residue. In another embodiment of the specified hydrophobic amino acid residue is achiral amino acid residue.

Used herein, the term "polar amino acid residue", unless otherwise specified, refers to a hydrophilic amino acid residue having a side chain that is uncharged at physiological pH, but which contains at least one link, in which a pair of electrons, socialized by two atoms is held in greater proximity to one of the atoms. Polar amino acid residues include, but are not limited to, Asn (N), Gln (Q), Ser (S)Thr (T), Cit, Pen, MSO, AcLys, hCys, hSer, Hyp, D-Asn, D-Gln, D-Ser, D-Thr, D-Cit, D-Pen, D-MSO, D-AcLys, D-hCys, D-hSer and D-Hyp. Other polar amino acids include, but are not limited to, analogs of the side chains C1-4with the following formula:

where n is an integer from 1 to 4. In one of the embodiments of the specified polar amino acid residue is L-amino acid residue. In another embodiment of the specified polar amino acid residue is a D-amino acid residue. In another embodiment of the specified polar amino acid residue is achiral amino acid residue.

Used here is Ermin "acidic amino acid residue", unless otherwise specified, refers to a hydrophilic amino acid residue with a pK value of the side chain, which is less than 7. Acidic amino acid residues usually at physiological pH values have negatively charged side chains due to the loss of the hydrogen ion. Acidic amino acid residues include, but are not limited to, Glu (E), Asp (D), D-Glu, D-Asp. Other acidic amino acid residues include, but are not limited to, analogs of the side chains C1-4having the following formula:

where n is an integer from 1 to 4. In one of the embodiments specified acidic amino acid residue is L-amino acid residue. In another embodiment of the specified acidic amino acid residue is a D-amino acid residue. In another embodiment of the specified polar amino acid residue is achiral amino acid residue.

Used herein, the term "basic amino acid residue", unless otherwise specified, refers to a hydrophilic amino acid residue with a pK value of the side chain, which is more than 7. Basic amino acid residues usually at physiological pH values have positively charged side chains due to the Association with the ion of gerania. Basic amino acid residues vkluchaut yourself not limited to, His (H), Arg (R)Lys (K), Dpr, Orn, hArg, Dbu, Dab, Phe(p-NH2), D-His, D-Arg, D-Lys, D-Dpr, D-Orn, D-hArg, D-Dbu, D-Dab, D-Phe(p-NH2). Other basic amino acid residues include, but are not limited to, analogs of the side chains C1-4with the following formula:

where n is an integer from 1 to 4. In one of the embodiments of the specified basic amino acid residue is L-amino acid residue. In another embodiment of the specified basic amino acid residue is a D-amino acid residue. In another embodiment of the specified basic amino acid residue is achiral amino acid residue.

Used herein, the term "nonpolar amino acid residue", unless otherwise specified, refers to a hydrophobic amino acid residue having a side chain that is uncharged at physiological pH and which contains, in which a pair of electrons, socialized by two atoms is held essentially to the same extent both of these atoms (i.e. the specified side chain is non-polar). Non-polar amino acid residues include, but are not limited to, Leu (L), Val (V), Ile (I), Met (M), Gly (G)Ala (A), Pro (P), azPro, Pip, azPip, β-Ala, Nip, t-BuG, MeIle, ChA, Nle, MeVal, hPro, 3-aminopropionic acid, 4-aminobutyric acid, Inp, Aib, Aha, Ava, MeGly, D-Leu, D-Val, D-le, D-Met, D-Ala, D-Pro, D-β-Ala, D-Inp-D-t-BuG, D-MeIle, D-ChA, D-Nle, D-MeVal, D-Nip, D-Pip and D-hPro. Other non-polar amino acid residues include, but are not limited to, analogs of the side chains C1-4with the following formula:

where n is an integer from 1 to 4. In one of the embodiments of the specified non-polar amino acid residue is L-amino acid residue. In another embodiment of the specified non-polar amino acid residue is a D-amino acid residue. In another embodiment of the specified non-polar amino acid residue is achiral amino acid residue.

Used herein, the term "aromatic amino acid residue", unless otherwise specified, refers to a hydrophobic amino acid residue with a side chain having at least one aromatic or heteroaromatic ring. Specified aromatic or heteroaromatic ring may contain one or more substituents such as-OH, -SH, -CN, -F, -Cl, -Br, -I, -NO2, -NO, -NH2, -Other, -NRR, -C(O)R, -C(O)OH, -C(O)OR, -C(O)NH2, -C(O)other, -C(O)NRR where each R independently is a (C1-C6)-alkyl, substituted (C1-C6)-alkyl, 5-26-membered aryl and substituted 5-26-membered aryl. Aromatic amino acid residues include, but are not limited to, Phe (F), Tyr (Y), Trp (W), PhG, Nal, Phe(4-Cl), Phe(2-F)Ph(3-F), Phe(4-F), Tic, Thi, hPhe, D-Phe, D-Tyr, D-Trp, D-PhG, D-Nal, D-Phe(4-Cl), D-Phe(2-F), D-Phe(3-F), D-Phe(4-F), D-Tic, D-Thi, D-hPhe. Other aromatic amino acid residues include, but are not limited to, analogs of the side chains C1-4with the following formula:

,

where n is an integer from 1 to 4. In one of the embodiments of the specified aromatic amino acid residue is L-amino acid residue. In another embodiment of the specified aromatic amino acid residue is a D-amino acid residue. In another embodiment of the specified aromatic amino acid residue is achiral amino acid residue.

Classification genetically encoded and non-genetically encoded amino acid residues in accordance with the above categories summarized in the table below. It should be understood that the table below is presented here only for illustrative purposes and does not purport to be an exhaustive list of amino acid residues that can be used in the here described mimetics ApoA-I. Other amino acid residues, especially not described here, can be easily attributed to a particular category based on the observed physical and chemical properties, taking into account the above definitions. Some classification of amino acid residues included in tab the Itza 2 below.

Table 2
Classification of some amino acid residues
ClassificationGenetically encodedNot genetically encoded
Hydrophobic
AromaticF, Y, WPhG, Nal, Thi, Tic, Phe(4-Cl), Phe(2-F), Phe(3-F), Phe(4-F), hPhe
NonpolarL, V, I, M, G, A, Pβ-Ala, t-BuA, t-BuG, MeIle, Nle, MeVal, ChA, MeGly, Aib, Nip, hPro, 3-aminopropionic acid, 4-aminobutyric acid, Inp, Aha, Ava, MeGly, azPro, Pip, azPip
AliphaticA, V, L, I, Pβ-Ala, Aib, t-BuA, t-BuG, MeIle, ChA, Nle, MeVal, Nip, hPro, Inp, azPro, Pip, azPip
Hydrophilic
SourD, E
MainH, K, RDpr, Orn, hArg, Phe(p-NH2), Dbu, Dab, hArg

PolarC, Q, N, S, TCit, Pen, AcLys, MSO, bAla, hSer, hCys, hSer, Hyp
Violating helical structureP, GMeGly, L-amino acids (D-peptides), D-Pro, and other D-amino acids (L-peptides)

As should be obvious to experts in this field, as defined above categories are not mutually exclusive. Thus, amino acid residues having a side chain exhibiting two or more physico-chemical properties, can be included in multiple categories. For example, amino acid side chain having an aromatic fragments, which are optionally substituted by polar substituents, such as Tyr (Y) or its corresponding D-enantiomer, can be as hydrophobic aromatic properties, and polar or hydrophilic properties, and therefore can be included in both categories, aromatic and polar. The assignment of any amino acid residue to one or another category should be obvious to the person skilled in the art, particularly in light of the present description.

In addition, the amino acid residue Cys (C) or its corresponding D-enantiomer can form disulfide bridges with other residues Cys (C) or sootvetstvuyuschimi D-enantiomers or other effect-free remedy containing amino acids. The ability residues Cys (C) (and other amino acids, containing SH-lateral chain) to exist in the peptide either in the reduced form with free SH-groups, either in the form of oxidized forming disulfide bridges form, affects whether residues Cys (C) or their respective D-enantiomers in the result, to give the peptide hydrophobic or hydrophilic character. Because Cys (C) or its corresponding D-enantiomer exhibits hydrophobicity, corresponding to 0.29, according to the conventional normalized scale of Eisenberg (Eisenberg, 1984, above), it should be understood that in order to implement the present invention Cys (C) and its corresponding D-enantiomer fall under the category of polar hydrophilic amino acids, despite the General classification, as defined above.

II. The mimic ApoA-I

A. Peptides of formula I

In one of the embodiments of the present invention is associated with peptides, containing from 15 to 29 residues having the formula I:

R1-Y1-X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-X18-X19-X20-X21-X22-X23-Y2-R2

The formula I

and their pharmaceutically acceptable salts, in which

X1is missing or achiral, D - or L-primary and inability residue;

X2is achiral, D -, or L-basic amino acid residue;

X3is achiral, D -, or L-aliphatic amino acid residue;

X4is achiral, D -, or L-basic amino acid residue;

X5represents Gln, Asn, D-Gln, D-Asn or achiral, D -, or L-basic amino acid residue;

X6represents Gln, Asn, D-Gln, D-Asn or achiral, D -, or L-basic amino acid residue;

X7is achiral, D -, or L-hydrophobic amino acid residue;

X8is achiral, D -, or L-hydrophobic amino acid residue;

X9is achiral, D -, or L-hydrophilic amino acid residue;

X10represents Leu, Trp, Gly, Nal, D-Leu, D-Trp or D-Nal;

X11represents Gly or achiral, D -, or L-aliphatic amino acid residue;

X12is achiral, D -, or L-hydrophilic amino acid residue;

X13is achiral, D -, or L-hydrophilic amino acid residue;

X14represents Leu, Trp, Gly, D-Leu, or D-Trp;

X15represents Leu, Gly or D-Leu;

X16is achiral, D -, or L-acidic amino acid residue, or achiral, D -, or L-basic amino acid residue;

X17is achiral, D -, or L-hydrophilic amino acid residue;

X18p is ecstasy a Leu, Phe, D-Leu or D-Phe;

X19represents Leu, Phe, D-Leu or D-Phe;

X20is achiral, D -, or L-acidic amino acid residue;

X21represents Leu, Phe, D-Leu or D-Phe;

X22is achiral, D -, or L-aliphatic amino acid residue; and

X23represents Inp, Nip, azPro, Pip, azPip, D-Nip or D-Pip;

Y1is absent or is an amino acid sequence containing from 1 to 7 residues;

Y2is absent or is an amino acid sequence containing from 1 to 7 residues;

R1represents H or aminosidine group;

R2represents OH or a carboxyl protective group;

moreover, from zero to eight residues X2-X22absent; and

thus

a) each chiral amino acid residue is L-amino acid residue;

b) each chiral amino acid residue is a D-amino acid residue;

(c) each chiral amino acid residue is L-amino acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is a D-amino acid residue; or

d) each chiral amino acid residue is a D-amino acid residue, excluding the rising, that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is an L-amino acid residue.

In another embodiment of the present invention is associated with peptides containing from 22 to 29 residues having the formula I:

R1-Y1-X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-X18-X19-X20-X21-X22-X23-Y2-R2

The formula I

and their pharmaceutically acceptable salts, in which

X1is missing or achiral, D -, or L-basic amino acid residue;

X2is achiral, D -, or L-basic amino acid residue;

X3is achiral, D -, or L-aliphatic amino acid residue;

X4is achiral, D -, or L-basic amino acid residue;

X5represents Gln, Asn, D-Gln, D-Asn or achiral, D -, or L-basic amino acid residue;

X6is achiral, D -, or L-basic amino acid residue;

X7is achiral, D -, or L-hydrophobic amino acid residue;

X8is achiral, D -, or L-hydrophobic amino acid residue;

X9is and what isalnum, D - or L-hydrophilic amino acid residue;

X10represents Leu, Trp, Gly, Nal, D-Leu, D-Trp or D-Nal;

X11represents Gly or achiral, D -, or L-aliphatic amino acid residue;

X12is achiral, D -, or L-hydrophilic amino acid residue;

X13is achiral, D -, or L-hydrophilic amino acid residue;

X14represents Leu, Trp, Gly, D-Leu, or D-Trp;

X15represents Leu, Gly or D-Leu;

X16is achiral, D -, or L-acidic amino acid residue;

X17is achiral, D -, or L-hydrophilic amino acid residue;

X18represents Leu, Phe, D-Leu or D-Phe;

X19represents Leu, Phe, D-Leu or D-Phe;

X20is achiral, D -, or L-acidic amino acid residue;

X21represents Leu, Phe, D-Leu or D-Phe;

X22is achiral, D -, or L-aliphatic amino acid residue; and

X23represents Inp, Nip, azPro, Pip, azPip, D-Nip or D-Pip;

Y1is absent or is an amino acid sequence containing from 1 to 7 residues;

Y2is absent or is an amino acid sequence containing from 1 to 7 residues;

R1represents H or aminosidine group;

R2represents OH or carboxylic zashitu the group; thus

a) each chiral amino acid residue is L-amino acid residue;

b) each chiral amino acid residue is a D-amino acid residue;

(c) each chiral amino acid residue is L-amino acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is a D-amino acid residue; or

d) each chiral amino acid residue is a D-amino acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is an L-amino acid residue.

In another embodiment of the present invention is associated with peptides, containing from 15 to 21 residue having the formula I:

R1-Y1-X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-X18-X19-X20-X21-X22-X23-Y2-R2

The formula I

and their pharmaceutically acceptable salts, in which

X1is missing or achiral, D -, or L-basic amino acid residue;

X2

X3is achiral, D -, or L-aliphatic amino acid residue;

X4is achiral, D -, or L-basic amino acid residue;

X5represents Gln, Asn, D-Gln, D-Asn or achiral, D -, or L-basic amino acid residue;

X6is achiral, D -, or L-basic amino acid residue;

X7is achiral, D -, or L-hydrophobic amino acid residue;

X8is achiral, D -, or L-hydrophobic amino acid residue;

X9is achiral, D -, or L-hydrophilic amino acid residue;

X10represents Leu, Trp, Gly, Nal, D-Leu, D-Trp or D-Nal;

X11represents Gly or achiral, D -, or L-aliphatic amino acid residue;

X12is achiral, D -, or L-hydrophilic amino acid residue;

X13is achiral, D -, or L-hydrophilic amino acid residue;

X14represents Leu, Trp, Gly, D-Leu, or D-Trp;

X15represents Leu, Gly or D-Leu;

X16is achiral, D -, or L-acidic amino acid residue;

X17is achiral, D -, or L-hydrophilic amino acid residue;

X18represents Leu, Phe, D-Leu or D-Phe;

X19represents Leu, Phe, D-Leu or D-Phe;

X20is achiral, D -, or L-acidic amino acid residue;

X21represents Leu, Phe, D-Leu or D-Phe;

X22is achiral, D -, or L-aliphatic amino acid residue; and

X23represents Inp, Nip, azPro, Pip, azPip, D-Nip or D-Pip;

Y1is absent or is an amino acid sequence containing from 1 to 7 residues;

Y2is absent or is an amino acid sequence containing from 1 to 7 residues;

R1represents H or aminosidine group;

R2represents OH or a carboxyl protective group;

and from one to eight residues X2-X22absent; and

a) each chiral amino acid residue is L-amino acid residue;

b) each chiral amino acid residue is a D-amino acid residue;

(c) each chiral amino acid residue is L-amino acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is a D-amino acid residue; or

d) each chiral amino acid residue is a D-amino acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues, right near St is only adjacent to them, is the L-amino acid residue.

In another embodiment, the peptide of formula I or its pharmaceutically acceptable salt has a 22 amino acid residue in length, and X1is missing.

Unless otherwise agreed, with imitators ApoA-I of formula I have the following implementation.

In one of the embodiments of the X2and X4both are Lys, Orn, D-Lys or D-Orn. In another embodiment X5is a GIn, Lys, D-Gln or D-Lys. In another embodiment X9is an acidic amino acid residue. In another embodiment X12represents Glu, Asn, Gln, Arg, D-Glu, D-Asn, D-Gln or D-Arg. In another embodiment X13represents Glu, Asn, Gln, Arg, D-Glu, D-Asn, D-Gln or D-Arg. In another embodiment X16is an acidic amino acid residue. In another embodiment X17represents Arg, Lys, Orn, D-Arg, D-Lys or D-Orn. In another embodiment X21represents aLeu or D-Leu. In another embodiment X22represents aAla, Val, Leu, D-Ala, D-Val or D-Leu.

In another embodiment X1is absent; X13is an acidic amino acid residue, Arg or D-Arg; X14is a basic amino acid residue, Asn, Glu, D-Asn or D-Glu; and X2-X12and X15-X23are as described above in formula I.

In another embodiment X1is absent; X2is Lys, Orn, D-Lys or D-Orn; X3is eu or D-Leu; X4is Lys, Orn, D-Lys or D-Orn; X5is Lys, Orn, Gln, Asn, D-Lys, D-Orn, D-Gln or D-Asn; X6represents Lys, Orn, Gln, Asn, D-Lys, D-Orn, D-Gln or D-Asn; X7represents Leu, Gly, Nal, D-Leu or D-Nal; X8represents Ala, Trp, Gly, Leu, Phe, Nal, D-Ala, D-Trp, D-Leu, D-Phe or D-Nal; X9represents Asp, Glu, Gln, Lys, D-Asp, D-Glu, D-Gln or D-Lys; X11represents Leu, Gly, D-Leu, or Aib; X12represents Asp, Glu, Asn, D-Asp, D-Glu or D-Asn; X13represents Asn, Gln, Glu, Arg, D-Asn, D-Gln, D-Glu or D-Arg; X16represents Asp, Arg, Glu, D-Asp, D-Arg or D-Glu; X17represents Lys, Arg, Orn, Asn, Glu, D-Lys, D-Arg, D-Orn, D-Asn or D-Glu; X20represents Asp, Glu, D-Asp or D-Glu; and/or X22represents Ala, Val, Leu, D-Ala, D-Val or D-Leu; and X10X14X15X18X19X21and X23are as defined above in formula I.

In another embodiment X1is absent; X9represents Glu or D-Glu; X12represents Glu or D-Glu; X13represents Asn, Glu, D-Asn or D-Glu; X14represents Leu or D-Leu; X15represents Leu or D-Leu; X16represents Glu or D-Glu; X17represents Arg, Lys, D-Arg or D-Lys; X18represents Phe or D-Phe; X19represents Leu or D-Leu; X21represents Leu or D-Leu; and/or X22represents Val or D-Val; and X2-X X10X11X20and X23are as defined above in formula I.

In another embodiment X1is absent; X2represents Lys, Orn, D-Lys or D-Orn; X3represents Leu or D-Leu; X4represents Lys, Orn, D-Lys or D-Orn; X5represents Lys, Orn, Gln, Asn, D-Lys, D-Orn, D-Gln or D-Asn; X6represents Lys, Orn, Gln, Asn, D-Lys, D-Orn, D-Gln or D-Asn; X7represents Leu, Gly, Nal, D-Leu or D-Nal; X8represents Ala, Trp, Gly, Leu, Phe, Nal, D-Ala, D-Trp, D-Leu, D-Phe or D-Nal; X9represents Glu or D-Glu; X11represents Leu, D-Leu, Gly, or Aib; X12represents Glu or D-Glu; X13represents Asn, Glu, D-Asn or D-Glu; X14represents Leu or D-Leu; X15represents Leu or D-Leu; X16represents Glu or D-Glu; X17represents Arg, Lys, D-Arg or D-Lys; X18represents Phe or D-Phe; X19represents Leu or D-Leu; X20represents Asp, Glu, D-Asp or D-Glu; X21represents Leu or D-Leu; and/or X22represents Val or D-Val; and X10and X23are as defined above in formula I.

In another embodiment X1no, only one of X5and X6is a basic amino acid residue, and the other of X5and X6is Gln, Asn, D-Gln or D-Asn.

In another embodiment Y1or Y 2is absent or is a sequence containing from one to seven amino acid residues. In another embodiment, one or more of the amino acid residues specified amino acid sequences are acidic amino acid residues. In another embodiment, one or more of the amino acid residues specified amino acid sequences are basic amino acid residues.

In another embodiment one of X5and X6is Lys, Orn, D-Lys or D-Orn, and the other of X5and X6is Gln, Asn, D-Gln or D-Asn.

In another embodiment each chiral amino acid residue is L-amino acid residue.

In another embodiment each chiral amino acid residue is a D-amino acid residue.

In another embodiment X1no; one of X7X8X10X11X14and X15is Gly; and X1-X6X9X12X13and X16-X23are other than Gly.

In another embodiment X1is absent; X11is Gly; and each of X7X8X10X14and X15is other than Gly.

In another embodiment X1is absent; X2represents Lys, Orn, D-Lys or D-Orn; X3represents Leu or D-Leu; X4is Lys, Orn, D-Lys or D-Orn; X5JW is aetsa Gln or D-Gln; X6represents Lys, Orn, D-Lys or D-Orn; X7represents Leu, Nal, D-Leu or D-Nal; X8is Ala, Trp, D-Ala or D-Trp; X9is Glu or D-Glu; X10represents Leu or D-Leu; X11is Gly; X12represents Glu or D-Glu; X13represents Asn or D-Asn; X14represents Leu, Trp, D-Leu, or D-Trp; X15is Leu or D-Leu; X16represents Glu or D-Glu; X17is Arg or D-Arg; X18represents Phe or D-Phe; X19represents Leu, Phe, D-Leu or D-Phe; X20represents Asp, Glu, D-Asp or D-Glu; X21is Leu or D-Leu; X22represents Val or D-Val; and X23is Inp. In one embodiments, the peptide of formula I is a peptide shown in table 3 below:

or their pharmaceutically acceptable salt.

In another embodiment X1is absent; X15is Gly; and each of X7X8X10X11and X14is other than Gly. In one embodiments, the peptide of formula I is as follows:

Peptide 10Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Gly-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:10); or
Peptide 12 Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Gly-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:102)

Peptide 222Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Gly-Glu-Arg-Phe-Leu-Asp-Leu-Val-azPro (SEQ ID NO:222)
Peptide 314Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Gly-Glu-Arg-Phe-Leu-Asp-Leu-Val-Pip (SEQ ID NO:314)
Peptide 406Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Gly-Glu-Arg-Phe-Leu-Asp-Leu-Val-azPip (SEQ ID NO:406)

or their pharmaceutically acceptable salt.

In another embodiment X1is absent; X14is Gly; and each of X7X8X10X11and X15is other than Gly. In one embodiments, the peptide of formula I is as follows:

Peptide 11Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Gly-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:11); or
Peptide 103Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Gly-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:103)
Peptide 223Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Gly-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-azPro (SEQ ID NO:223)
Peptide 315Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-sn-Gly-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Pip (SEQ ID NO:315)

Peptide 407Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Gly-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-azPip (SEQ ID NO:407)

or their pharmaceutically acceptable salt.

In another embodiment X1is absent; X10is Gly; and each of X7X8X11X14and X15is other than Gly. In one embodiments, the peptide of formula I is as follows:

Peptide 12Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Gly-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:12); or
Peptide 104Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Gly-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:104)
Peptide 224Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Gly-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-azPro (SEQ ID NO:224)
Peptide 316Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Gly-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Pip (SEQ ID NO:316)
Peptide 408Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Gly-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-azPip (SEQ ID NO:408)

or their pharmaceutically acceptable salt.

In another embodiment X1is absent; X8is Gly; and each of X7X10X11X14and X5 is other than Gly. In one embodiments, the peptide of formula I is as follows:

Peptide 13Lys-Leu-Lys-Gln-Lys-Leu-Gly-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:13); or
Peptide 105Lys-Leu-Lys-Gln-Lys-Leu-Gly-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:105)
Peptide 225Lys-Leu-Lys-Gln-Lys-Leu-Gly-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-azPro (SEQ ID NO:225)
Peptide 317Lys-Leu-Lys-Gln-Lys-Leu-Gly-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Pip (SEQ ID NO:317)
Peptide 409Lys-Leu-Lys-Gln-Lys-Leu-Gly-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-azPip (SEQ ID NO:409)

or their pharmaceutically acceptable salt.

In another embodiment X1is absent; X7is Gly; and each of X8X10X11X14and X15is other than Gly. In one embodiments, the peptide of formula I is as follows:

Peptide 14Lys-Leu-Lys-Gln-Lys-Gly-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:14); or
Peptide 106Lys-Leu-Lys-Gln-Lys-Gly-Ala-Glu-Leu-Leu-Gl-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:106)

Peptide 226Lys-Leu-Lys-Gln-Lys-Gly-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-azPro (SEQ ID NO:226)
Peptide 318Lys-Leu-Lys-Gln-Lys-Gly-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Pip (SEQ ID NO:318)
Peptide 410Lys-Leu-Lys-Gln-Lys-Gly-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-azPip (SEQ ID NO:410)

or their pharmaceutically acceptable salt.

In another embodiment X1is absent; and each of X7,X8X10X11X14and X15is other than Gly.

In another embodiment X1is absent; X2represents Lys, Orn, D-Lys or D-Orn; X3represents Leu or D-Leu; X4is Lys, Orn, D-Lys or D-Orn; one of X5or X6is Gln or D-Gln, and the other of X5or X6represents Lys, Orn, D-Lys or D-Orn; X7represents Leu, Nal, D-Leu or D-Nal; X8is Ala, Leu, Trp, Nal, D-Ala, D-Leu, D-Trp or D-Nal; X9is Glu or D-Glu; X10represents Leu, Trp, Nal, D-Leu, D-Trp or D-Nal; X11is Leu, D-Leu, or Aib; X12represents Glu or D-Glu; X13represents Asn, Gln, D-Asn or D-Gln; X14represents Leu, Trp, D-Leu, or D-Trp; X15is Leu or D-Leu; X16represents Glu or D-Glu; X is Arg, Lys, D-Arg or D-Lys; X18represents Leu, Phe, D-Leu or D-Phe; X19represents Leu, Phe, D-Leu or D-Phe; X20represents Asp, Glu, D-Asp or D-Glu; X21is Leu or D-Leu; X22represents Val, Leu, D-Val or D-Leu; and X23is Inp.

In another embodiment X1is absent; X2represents Lys or D-Lys; X3represents Leu or D-Leu; X4is Lys or D-Lys; X5represents Gln or D-Gln; X6is Lys or D-Lys; X7represents Leu or D-Leu; X8is Ala or D-Ala; X9is Glu or D-Glu; X10represents Leu or D-Leu; X11is Leu or D-Leu; X12represents Glu or D-Glu; X13represents Asn or D-Asn; X14represents Leu or D-Leu; X15is Leu or D-Leu; X16represents Glu or D-Glu; X17is Arg or D-Arg; X18represents Phe or D-Phe; X19represents Leu or D-Leu; X20represents Asp or D-Asp; X21is Leu or D-Leu; X22represents Val or D-Val; and X23represents Inp.

In another embodiment X3is other than Lys or D-Lys; X9is other than Trp or D-Trp; X11is different from Glu, Trp, D-Glu or D-Trp; X12is other than Trp, Leu, D-Trp or D-Leu; X13is other than Trp or D-Trp; X15is the I other than Lys, Trp, D-Lys or D-Trp; X17is other than Trp, Leu, D-Trp or D-Leu; X18is other than Trp or D-Trp; X19is other than Lys, Glu, Trp, Nal, D-Lys, D-Glu, D-Trp or D-Nal; and/or X22is different from Val or D-Val.

In another embodiment, the peptide of formula I is one of the peptides shown in table 4 below:

or their pharmaceutically acceptable salt.

B.The peptides of formula II

In one of the embodiments of the present invention is associated with peptides, containing from 14 to 22 residues having the following formula II:

R1-Y1-X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-X18-Y2-R2

Formula II

and their pharmaceutically acceptable salts, in which

X1is achiral, D -, or L-basic amino acid residue;

X2represents Leu or D-Leu;

X3is achiral, D -, or L-basic amino acid residue;

X4represents Gln, Asn, D-Gln or D-Asn;

X5p is ecstasy a Leu, D-Leu or achiral, D -, or L-basic amino acid residue;

X6represents Leu, Trp, Phe, D-Leu, D-Trp or D-Phe;

X7is achiral, D -, or L-acidic amino acid residue;

X8represents Asn, D-Asn or is achiral, D -, or L-acidic amino acid residue;

X9represents Leu, Trp, D-Leu, or D-Trp;

X10represents Leu, Trp, D-Leu, or D-Trp;

X11is an achiral, D -, or L-acidic amino acid residue;

X12is achiral, D -, or L-basic amino acid residue;

X13represents Leu, Phe, D-Leu or D-Phe;

X14represents Leu, Phe, D-Leu or D-Phe;

X15is achiral, D -, or L-acidic amino acid residue;

X16represents Leu or D-Leu;

X17is achiral, D -, or L-aliphatic amino acid residue;

X18represents Inp, Nip, azPro, Pip, azPip, D-Nip or D-Pip;

Y1is absent or is an amino acid sequence containing from 1 to 4 residues;

Y2is absent or is an amino acid sequence containing from 1 to 4 residues;

R1represents H or aminosidine group;

R2represents OH or a carboxyl protective group;

moreover, from zero to eight residues from the X1to X17no the are; and

a) each chiral amino acid residue is L-amino acid residue;

b) each chiral amino acid residue is a D-amino acid residue;

(c) each chiral amino acid residue is L-amino acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is a D-amino acid residue; or

d) each chiral amino acid residue is a D-amino acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is an L-amino acid residue.

In another embodiment of the present invention is associated with peptides, containing from 15 to 22 residues having the following formula II:

R1-Y1-X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-X18-Y2-R2

Formula II

and their pharmaceutically acceptable salts, in which

X1is achiral, D -, or L-basic amino acid residue;

X2represents Leu or D-Leu;

X3one is by achiral, D - or L-basic amino acid residue;

X4represents Gln, Asn, D-Gln or D-Asn;

X5represents Leu, D-Leu, or is achiral, D -, or L-basic amino acid residue;

X6represents Leu, Trp, Phe, D-Leu, D-Trp or D-Phe;

X7is achiral, D -, or L-acidic amino acid residue;

X8represents Asn, D-Asn or is achiral, D -, or L-acidic amino acid residue;

X9represents Leu, Trp, D-Leu, or D-Trp;

X10represents Leu, Trp, D-Leu, or D-Trp;

X11is an achiral, D -, or L-acidic amino acid residue;

X12is achiral, D -, or L-basic amino acid residue;

X13represents Leu, Phe, D-Leu or D-Phe;

X14represents Leu, Phe, D-Leu or D-Phe;

X15is achiral, D -, or L-acidic amino acid residue;

X16represents Leu or D-Leu;

X17is achiral, D -, or L-aliphatic amino acid residue;

X18represents Inp, Nip, azPro, Pip, azPip, D-Nip or D-Pip;

Y1is absent or is an amino acid sequence containing from 1 to 4 residues;

Y2no;

R1represents H or aminosidine group;

R2represents OH or a carboxyl protective group;

moreover, the t of the zero to three residues from the X 1to X17absent; and

a) each chiral amino acid residue is L-amino acid residue;

b) each chiral amino acid residue is a D-amino acid residue;

(c) each chiral amino acid residue is L-amino acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is a D-amino acid residue; or

d) each chiral amino acid residue is a D-amino acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is an L-amino acid residue.

In another embodiment of the present invention is associated with peptides containing 14 residues having the following formula II:

R1-Y1-X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-X18-Y2-R2

Formula II

and their pharmaceutically acceptable salts, in which

X1is achiral, D -, or L-basic amino acid residue;

X2represents Leu or DLeu;

X3is achiral, D -, or L-basic amino acid residue;

X4represents Gln, Asn, D-Gln or D-Asn;

X5represents Leu, D-Leu, or is achiral, D -, or L-basic amino acid residue;

X6represents Leu, Trp, Phe, D-Leu, D-Trp or D-Phe;

X7is achiral, D -, or L-acidic amino acid residue;

X8represents Asn, D-Asn or is achiral, D -, or L-acidic amino acid residue;

X9represents Leu, Trp, D-Leu, or D-Trp;

X10represents Leu, Trp, D-Leu, or D-Trp;

X11is an achiral, D -, or L-acidic amino acid residue;

X12is achiral, D -, or L-basic amino acid residue;

X13represents Leu, Phe, D-Leu or D-Phe;

X14represents Leu, Phe, D-Leu or D-Phe;

X15is achiral, D -, or L-acidic amino acid residue;

X16represents Leu or D-Leu;

X17is achiral, D -, or L-aliphatic amino acid residue;

X18represents Inp, Nip, azPro, Pip, azPip, D-Nip or D-Pip;

Y1is absent or is an amino acid sequence containing from 1 to 4 residues;

Y2no;

R1represents H or aminosidine group;

R2represents OH or a carboxy is inuu protective group;

and from four to eight residues from the X1to X17absent; and

a) each chiral amino acid residue is L-amino acid residue;

b) each chiral amino acid residue is a D-amino acid residue;

(c) each chiral amino acid residue is L-amino acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is a D-amino acid residue; or

d) each chiral amino acid residue is a D-amino acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is an L-amino acid residue.

In one of the embodiments a peptide of the formula II is a peptide containing 18 residues.

In one of the embodiments a peptide of the formula II is a peptide represented below in table 5.

or their pharmaceutically acceptable salt.

C. the Peptides of formula III

In one of the embodiments of the present invention is associated with peptides, containing from 15 to 29 residues, iluminatului III:

R1-Y1-X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-X18-X19-X20-X21-X22-X23-Y2-R2

Formula III

and their pharmaceutically acceptable salts, in which

X1is missing or achiral, D -, or L-basic amino acid residue;

X2is achiral, D -, or L-basic amino acid residue;

X3is achiral, D -, or L-aliphatic amino acid residue;

X4is achiral, D -, or L-basic amino acid residue;

X5represents Gln, Asn, D-Gln, D-Asn or is achiral, D -, or L-basic amino acid residue;

X6represents Gln, Asn, D-Gln or D-Asn, or is achiral, D -, or L-basic amino acid residue;

X7is achiral, D -, or L-hydrophobic amino acid residue;

X8is achiral, D -, or L-hydrophobic amino acid residue;

X9is achiral, D -, or L-hydrophilic amino acid residue;

X10represents aLeu, Trp, Gly, Nal, D-Leu, D-Trp or D-Nal;

X11represents Gly or is achiral, D -, or L-aliphatic amino acid residue;

X12is shall be Ahira enim, D - or L-hydrophilic amino acid residue;

X13is achiral, D -, or L-hydrophilic amino acid residue;

X14represents Leu, Trp, Gly, D-Leu, or D-Trp;

X15represents Leu, Gly or D-Leu;

X16is achiral, D -, or L-acidic amino acid residue or achiral, D -, or L-basic amino acid residue;

X17is achiral, D -, or L-hydrophilic amino acid residue;

X18represents Leu, Phe, D-Leu or D-Phe;

X19represents Leu, Phe, D-Leu or D-Phe;

X20is achiral, D -, or L-acidic amino acid residue;

X21represents Leu, Phe, D-Leu or D-Phe;

X22is achiral, D -, or L-aliphatic amino acid residue; and

X23represents Inp, Nip, azPro, Pip, azPip, D-Nip or D-Pip;

Y1is absent or is an amino acid sequence containing from 1 to 7 residues;

Y2is absent or is an amino acid sequence containing from 1 to 7 residues;

R1represents H or aminosidine group;

R2represents OH or a carboxyl protective group; and zero to eight residues X2-X22absent; and

a) each chiral amino acid residue is L-amino acid residue;

b) all the chiral amino acid residue is a D-amino acid residue;

(c) each chiral amino acid residue is L-amino acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is a D-amino acid residue; or

d) each chiral amino acid residue is a D-amino acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is an L-amino acid residue.

In another embodiment of the present invention is associated with peptides containing from 22 to 29 residues having the formula III:

R1-Y1-X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-X18-X19-X20-X21-X22-X23-Y2-R2

Formula III

and their pharmaceutically acceptable salts, in which

X1is missing or achiral, D -, or L-basic amino acid residue;

X2is achiral, D -, or L-basic amino acid residue;

X3is achiral, D -, or L-aliphatic amino acid residue;

X4is shall be Ahira enim, D - or L-basic amino acid residue;

X5represents Gln, Asn, D-Gln, D-Asn or is achiral, D -, or L-basic amino acid residue;

X6is achiral, D -, or L-basic amino acid residue;

X7is achiral, D -, or L-hydrophobic amino acid residue;

X8is achiral, D -, or L-hydrophobic amino acid residue;

X9is achiral, D -, or L-hydrophilic amino acid residue;

X10represents aLeu, Trp, Gly, Nal, D-Leu, D-Trp or D-Nal;

X11represents Gly or is achiral, D -, or L-aliphatic amino acid residue;

X12is achiral, D -, or L-hydrophilic amino acid residue;

X13is achiral, D -, or L-hydrophilic amino acid residue;

X14represents Leu, Trp, Gly, D-Leu, or D-Trp;

X15represents Leu, Gly or D-Leu;

X16is achiral, D -, or L-acidic amino acid residue;

X17is achiral, D -, or L-hydrophilic amino acid residue;

X18represents Leu, Phe, D-Leu or D-Phe;

X19represents Leu, Phe, D-Leu or D-Phe;

X20is achiral, D -, or L-acidic amino acid residue;

X21represents Leu, Phe, D-Leu or D-Phe;

X22is achiral, D - and L-aliphatic amino acid residue; and

X23represents Inp, Nip, azPro, Pip, azPip, D-Nip or D-Pip;

Y1is absent or is an amino acid sequence containing from 1 to 7 residues;

Y2is absent or is an amino acid sequence containing from 1 to 7 residues;

R1represents H or aminosidine group;

R2represents OH or a carboxyl protective group;

a) each chiral amino acid residue is L-amino acid residue;

b) each chiral amino acid residue is a D-amino acid residue;

(c) each chiral amino acid residue is L-amino acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is a D-amino acid residue; or

d) each chiral amino acid residue is a D-amino acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is an L-amino acid residue.

In another embodiment of the present invention is associated with peptides, containing from 15 to 21 residue having the formula III:

R1-Y1- 1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-X18-X19-X20-X21-X22-X23-Y2-R2

Formula III

and their pharmaceutically acceptable salts, in which

X1is missing or achiral, D -, or L-basic amino acid residue;

X2is achiral, D -, or L-basic amino acid residue;

X3is achiral, D -, or L-aliphatic amino acid residue;

X4is achiral, D -, or L-basic amino acid residue;

X5represents Gln, Asn, D-Gln, D-Asn or is achiral, D -, or L-basic amino acid residue;

X6is achiral, D -, or L-basic amino acid residue;

X7is achiral, D -, or L-hydrophobic amino acid residue;

X8is achiral, D -, or L-hydrophobic amino acid residue;

X9is achiral, D -, or L-hydrophilic amino acid residue;

X10represents aLeu, Trp, Gly, Nal, D-Leu, D-Trp or D-Nal;

X11represents Gly or is achiral, D -, or L-aliphatic amino acid residue;

X12is achiral, D -, or L-hydrophilic amino acid residue;

X13

X14represents Leu, Trp, Gly, D-Leu, or D-Trp;

X15represents Leu, Gly or D-Leu;

X16is achiral, D -, or L-acidic amino acid residue;

X17is achiral, D -, or L-hydrophilic amino acid residue;

X18represents Leu, Phe, D-Leu or D-Phe;

X19represents Leu, Phe, D-Leu or D-Phe;

X20is achiral, D -, or L-acidic amino acid residue;

X21represents Leu, Phe, D-Leu or D-Phe;

X22is achiral, D -, or L-aliphatic amino acid residue; and

X23represents Inp, Nip, azPro, Pip, azPip, D-Nip or D-Pip;

Y1is absent or is an amino acid sequence containing from 1 to 7 residues;

Y2is absent or is an amino acid sequence containing from 1 to 7 residues;

R1represents H or aminosidine group;

R2represents OH or a carboxyl protective group; and from one to eight residues X2-X22absent; and

a) each chiral amino acid residue is L-amino acid residue;

b) each chiral amino acid residue is a D-amino acid residue;

(c) each chiral amino acid residue is L-Amin is an acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is a D-amino acid residue; or

d) each chiral amino acid residue is a D-amino acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is an L-amino acid residue.

In another embodiment, the peptide of formula III contains 22 amino acid residue in length, and X1is missing.

In one embodiments, the peptide of formula III is a peptide represented below in table 6.

or their pharmaceutically acceptable salt.

In other embodiments of the present invention includes a mimic ApoA-I, in which one or more of their amide linkages optionally replaced by a linkage other than amide, including, but not limited to, substituted amidon or isosteric amide. Thus, although the various remnants of the X1-X23, Y1and Y2in formulas I, II and III are described in terms of amino acids, in particular in the lewinian of the present invention, instead of one or more amide bonds present nelena connection.

In another embodiment, the nitrogen atom one or more of the amide bonds mimetics of ApoA-I is substituted, that is substituted by an amide linkage has the formula-C(O)NR'-, where R' represents a (C1-C6)-alkyl, (C2-C6)-alkenyl, (C2-C6)-quinil, (C5-C20)-aryl, (C6-C26-aralkyl, 5-20-membered heteroaryl or 6-26 membered algeterror. In another embodiment R' is a substituted-OR, -SR, -NRR, -NO2, -CN, halogen, -SO2R, -C(O)R, -C(O)OR and-C(O)NRR where each R is independently hydrogen, alkyl or aryl.

In another embodiment nelena communication replaces one or more amide bonds mimetics of ApoA-I and includes, but is not limited to, -CH2NH-, -CH2S-, -CH2CH2-, -CH=CH- (CIS and TRANS), -C(O)CH2-, -CH(OH)CH2- and-CH2SO-. Compounds having such neamine communication, as well as methods for such compounds is well known to specialists in this field (see, for example, Spatola, March 1983, Vega Data Vol.1, Issue 3; Spatola, 1983, "Peptide Backbone Modifications" In: Chemistry and Biochemistry of Amino Acids Peptides and Proteins, Weinstein, ed., Marcel Dekker, New York, p.267 (review of a General nature); Morley, 1980, Trends Pharm. Sci. 1: 463-468; Hudson et al., 1979, Int. J. Prot. Res. 14: 177-185 (-CH2NH-, -CH2CH2-); Spatola et al., 1986, Life Sci. 38: 1243-1249 (-CH2-S); Hann, 1982, J. Chem. Soc. Perkin Trans. I. 1: 307-314 (-CH=CH-, CIS - and Tr is the na-); Almquist et al., 1980, J. Med. Chem. 23: 1392-1398 (-COCH2-); Jennings-White et al., Tetrahedron. Lett. 23: 2533 (-COCH2-); European Patent Application EP 45665 (1982) CA 97: 39405 (-CH(OH)CH2-); Holladay et al., 1983, Tetrahedron Lett. 24: 4401-4404 (-C(OH)CH2-); and Hruby, 1982, Life Sci. 31: 189-199 (-CH2-S-).

In addition, one or more amide bonds mimetics of ApoA-I can be replaced by one or more peptidomimetics or imitators amide fragments that do not cause significant violations of the structure or activity of these peptides. Suitable simulators amide fragments are described, for example, Olson et al., 1993, J. Med. Chem. 36: 3039-3049.

In certain embodiments, the mimetic ApoA-I is presented in the form of pharmaceutically acceptable salts. Salt can be formed on the C-end or at the N-end or side chain acidic or basic amino acid residue.

In certain embodiments of the pharmaceutically acceptable salt is a salt of the metal salt of the organic amine or salt accession acids.

Metal salt can be formed by adding an inorganic base to the peptide of formula I, II or III. Inorganic base consists of a metal cation, paired with a counterion of the base, such as hydroxide, carbonate, bicarbonate or phosphate. The metal can be alkali metal, alkaline earth metal, transition metal or a main group metal. ODA is divided embodiments the metal is lithium, sodium, potassium, cerium, magnesium, manganese, iron, calcium, aluminum or zinc.

Organic amine salts can be formed by adding an organic amine to the peptide of formula I, II or III. In certain embodiments the organic amine is triethylamine, ethanolamine, diethanolamine, triethanolamine, morpholine, piperidine, N-methylpiperidine, N-ethylpiperidine, dibenzylamine, piperazine, pyridine, pyrazin or piperazine.

Salt accession acids can be formed by adding acid to the peptide of formula I, II or III. In certain embodiments, the acid is organic. In certain embodiments, the acid is inorganic. In other embodiments, the acid is hydrochloric acid, Hydrobromic acid, itestosterone acid, nitric acid, sulfuric acid, sulfurous acid, phosphoric acid, isonicotinic acid, lactic acid, salicylic acid, tartaric acid, ascorbic acid, hentaimovi acid, gluconic acid, glucuronic acid, sugar acid, formic acid, benzoic acid, glutamic acid, Pantothenic acid, acetic acid, fumaric acid, succinic acid, methanesulfonic acid, econsultancy acid, benzosulfimide acid, para-toluensulfonate acid, citric KIS the Auteuil or maleic acid. In certain other embodiments the salt of the accession acids is hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, sulfite, bisulfate, phosphate, acid phosphate, isonicotinate, lactate, salicylate, tartrate, bitartrate, ascorbate, getitemat, gluconate, glucuronate, saharat, format, benzoate, glutamate, Pantothenate, acetate, fumarate, succinate, methanesulfonate, aconsultant, bansilalpet, para-toluensulfonate, citrate or malate.

In certain embodiments, R1represents aminosidine group. In certain embodiments specified aminosidine group are (C1-C6)-alkyl, (C2-C6)-alkenyl, (C2-C6)-quinil, (C5-C26)-aryl, (C6-C26-aralkyl), 5-20-membered heteroaryl or 6-26 membered algeterror; -C(O)R; -C(O)OR; -SO2R; or,- SR, where R is H or (C1-C6)-alkyl, (C2-C6-alkenyl, (C2-C6-quinil, (C5-C26)-aryl, (C6-C26-aralkyl 5-20-membered heteroaryl or 6-26 membered by achetercialis. In other embodiments (C1-C6)-alkyl, (C2-C6)-alkenyl, (C2-C6)-quinil, (C5-C26)-aryl, (C6-C26-aralkyl), 5-20-membered heteroaryl or 6-26 membered algeterror substituted by one or more ORa, -SRa, -NRaRa, -NO2, -CN, halogen,-SO 2Ra, -C(O)Ra, -C(O)ORaand-C(O)NRaRawhere each Raindependently is hydrogen, alkyl or aryl. When R1is H, the number aminosidine groups in the mimetic ApoA-I is zero; and when R1is aminosidine group, the number aminosidine groups in the mimetic ApoA-I is equal to 1.

In other embodiments aminosidine group are dansyl; methoxycarbonyl; etoxycarbonyl; 9-fertilitycare; 2-chlorocarbons; 2,2,2-trichlorocyanuric; 2-fenilalaninammonii; tert-butoxycarbonyl; benzyloxycarbonyl; para-methoxybenzyloxy; para-nitrobenzenesulfonyl;o- nitrobenzisoxazole; para-bromobenzyloxycarbonyl; para-chlorobenzenesulfonyl; para-identilication; 2,4-dichlorobenzenesulfonyl; diphenylmethylene; 3,5-dimethoxybenzonitrile; phenoxycarbonyl; 2,4,6-tri-tert-butylphenoxyacetyl; 2,4,6-trimethylbenzenesulfonyl; formyl; acetyl; chloroacetyl; trichloracetic; TRIFLUOROACETYL; phenylacetyl; pikolinos; benzoyl; para-phenylbenzyl; phthaloyl; methyl; tert-butyl; allyl; [2-(trimethylsilyl)ethoxy]methyl; 2,4-dimethoxybenzyl; 2,4-dinitrophenyl; benzyl; 4-methoxybenzyl; diphenylmethyl; triphenylmethyl; benzazolyl;o-nitrobenzenesulfenyl; 2,4-dinitrobenzenesulfonyl; para-toluensulfonyl; benzazolyl; 2,3,6-trimethyl-4-methoxybenzo sulfonyl; 2,4,6-trimethoxybenzaldehyde; 2,6-dimethyl-4-methoxybenzenesulfonyl; pentamethylbenzene; 4-methoxybenzenesulfonyl; 2,4,6-trimethylbenzenesulfonyl; or benzylmethyl. In other embodiments aminosidine group is acetyl, formyl or dansyl.

In certain embodiments, R2represents H or a carboxyl protective group. In certain embodiments of the carboxyl protecting group is O-(C1-C6)-alkyl, O-(C2-C6)-alkenyl, O-(C2-C6)-quinil, O-(C5-C26)-aryl, O-(C5-C26-aralkyl), O-(5-20-membered heteroaryl) or O-(6-26-membered algeterror); or-NRR where R represents H or (C1-C6)-alkyl, (C2-C6)-alkenyl, (C2-C6)-quinil, (C5-C26)-aryl, (C6-C26-aralkyl), 5-20-membered heteroaryl or 6-26 membered algeterror. In other embodiments (C1-C6)-alkyl, (C2-C6)-alkenyl, (C2-C6)-quinil, (C5-C26)-aryl, (C6-C26-aralkyl, 5-20-membered heteroaryl or 6-26 membered algeterror substituted by one or more from ORa, -SRa, -NRaRa, -NO2, -CN, halogen, -SO2Ra, -C(O)Ra, -C(O)ORaand-C(O)NRaRawhere each Raindependently is hydrogen, alkyl or aryl. When R1is H, the number of ka is boxylic protective groups in mimetic ApoA-I is equal to zero; and when R1is a carboxyl protecting group, the number of carboxyl protective groups in mimetic ApoA-I is equal to 1.

In other embodiments carboxyl protective group are methoxy; ethoxy; 9 fertilitate; methoxyethoxy; methylthiomethyl; tetrahydropyranyloxy; tetrahydrofuranate; methoxyethoxyethoxy; benzylacetone; phenacyloxy; pair-bromination; α-methylbenzylamine; pair-methoxybenzyloxy; depilarsi; 2 chloroethoxy; 2,2,2-trichloroethane, 2-methylthioethyl; 2-(para-toluensulfonyl)methoxy; tert-butoxy; cyclopentane; cyclohexane; allyloxy; metalliance; cinnamate; α-methylcinnamic; phenoxy; 2,6-dimethylphenoxy; 2,6-diisopropylphenol; benzyloxy; triphenylmethane; diphenylmethoxy; 2,4,6-trimethylsiloxy; pair-bromobenzylamine;o-nitrobenzyloxy; N,N-dimethylamide; pyrrolidinyl; or piperidinyl.

In the scope of the present invention also includes a protected form mimetics of ApoA-I, i.e. forms mimetics of ApoA-I in which one or more of them-NH2- or COOH-group protected by a protective group. In one embodiments, one or more-NH2groups protected aminosidine group, as described above. In another embodiment, one or more COOH groups are protected, as described above, the carboxyl protecting group.

In one of the embodiments of the mimic ApoA-I have the ability obrazovyvat the amphipatic α-helix in the presence of one or more lipids. The term "amphipatic, or amphiphilic" means that this α-helix is the opposite of hydrophilic and hydrophobic surfaces, oriented along its long axis, that is, on one side of this spiral projected mainly hydrophilic side chains, whereas on the opposite side of the spiral projected mostly hydrophobic side chains. On figa and 1B presents two illustrative image the opposite of hydrophilic and hydrophobic sides idealized model amphipatic α-helix. On figa presents chart Schiffer-Edmundson "helical wheel" ("Helical wheel") (Schiffer and Edmundson, 1967, Biophys. J. 7: 121-135). In this "wheel" long helix axis perpendicular to the page. Starting from the N-Terminus, consecutive amino acid residues (represented by circles) are radially located around the perimeter of the ring at intervals of 100°. Thus, the amino acid residue n+1 is the angle of rotation 100° relative to the residue n, n+2 is the angle of rotation 100° relative to the residue n+1, etc. the location of the angle of rotation 100° results in a 3.6 amino acid residue on the coil, which is usually observed in idealized α-helix. On figa clearly the opposite of hydrophilic and hydrophobic surface of the spiral; hydrophilic amino acid OS is atki denoted by hollow circles, and the filled circles are hydrophobic amino acid residues.

On FIGU presents the schematic diagram of the spiral grid idealized amphipatic helix depicted in figa. (Lim, 1978, FEBS Lett. 89: 10-14). In a typical circuit diagram of the spiral mesh α-helix is represented in the form of a cylinder which is cut along the center of its hydrophilic surface and straightened. Thus, the Central hydrophobic surfaces defined hydrophobic moment of the helix (Eisenberg et al., 1982, Nature 299: 371-374), lies in the center of this shape and oriented in such a way that out of the plane of the page. Illustration of a spiral cylinder to the cutting and correcting shown in figs. By cutting the cylinder in different planes gives the opportunity to observe different patterns of the same aliphatic spiral, and in this way you can get different information about the properties of the spiral.

Although the inventors do not wish to bind themselves to any particular theory, however, they believe that certain structural and/or physical properties of the amphipatic helix formed mimic ApoA-I may be important for their activity. Such properties include the degree of amphiphilicity, the total hydrophobicity, the average hydrophobicity, hydrophobic and hydrophilic corners, hydrophobic moment, the average hydrophob the first time, and the total charge of the α-helix.

The degree of amphiphilicity (the degree of asymmetry of hydrophobicity) amphipatic helix formed mimic ApoA-I, it is convenient to count by calculating the hydrophobic moment (µH) spiral. The methods of calculating µHfor specific peptide sequences are well known in this field and are described, for example, Eisenberg, 1984, Ann. Rev. Biochem. 53: 595-623. The actual value of µHreceived for a particular peptide will depend on the total number of amino acid residues constituting the peptide. Normally, therefore, a direct comparison of the values of µHfor peptides of different lengths is not informative.

Affilinet peptides of different lengths can be directly compared at the mean hydrophobic moment (<µH>). The average hydrophobic moment can be obtained by dividing µHon the number of residues in the helix (i.e. <µH> = µH/N). Usually mimic ApoA-I, manifesting <µH> from 0.45 to 0.65, which is determined using standard normalized hydrophobicity scale of Eisenberg (Eisenberg, 1984, J. Mol. Biol. 179: 125-142), are included in the scope of the present invention. In one of the embodiments <µH> is in the region of from 0.50 to 0.60.

Aggregate, or total, hydrophobicity (Ho) peptide conveniently be calculated by taking the algebraic sum of gidrofobnosti every what about the amino acid residue in the peptide (i.e. Hopho=i=1NHi), where N corresponds to the number of amino acid residues in the peptide, and Hicorresponds to the hydrophobicity of the ith amino acid residue). The average hydrophobicity (<HO>) is hydrophobicity, divided by the number of amino acid residues (i.e. <HO>=HO/N). Usually mimic ApoA-I, which have an average hydrophobicity in the field from -0,050 to -0,070 determined using conventional normalized hydrophobicity scale of Eisenberg (Eisenberg, 1984, J. Mol. Biol. 179: 125-142), are included in the scope of the present invention. In one of the embodiments of the average hydrophobicity is found in the region from -0,030 to -0,055.

The total hydrophobicity of the hydrophobic surface (Hopho) amphipatic helix can be obtained by taking the sum of gidrofobnosti hydrophobic amino acid residues that fall within the scope of hydrophobic angle, as described below (that is,Hopho=i=1 NHi), where Hicorresponds to the definition given above, and NHrepresents the total number of hydrophobic amino acid residues on the hydrophobic surface). The average hydrophobicity hydrophobic surface (<Hopho>) is Hopho/NHwhere NHcorresponds to the above definition. Usually mimic ApoA-I, which have an average hydrophobicity <Hopho> from 0.90 to 1.20, which is defined using conventional normalized hydrophobicity scale of Eisenberg (Eisenberg, 1984, above; Eisenberg et al., 1982, above), are included in the scope of the present invention. In one of the embodiments of the average hydrophobicity <Hopho> is in the range from 0.94 to 1.10 is.

Hydrophobic angle (pho-angle) is usually defined as the angle or arc covered the longest continuous segment of hydrophobic amino acid residues, when the peptide consider according to Schiffer-Edmundson "helical wheel" ("Helical wheel") (i.e. the number of contiguous hydrophobic residues on the wheel multiplied by 20°). Hydrophilic angle (phi-angle) represents the difference between 360° pho-angle (i.e. 360° pho-angle). The specialist is m in this area should be clear, that pho and phi-angles can partly dependent on the number of amino acid residues in the peptide. For example, with reference to figure 2, one can see that only 18 amino acid residues suitable for one turn spiral "wheels" Schiffer-Edmundson to 22-membered consensus peptide (Segrest''s consensus 22-mer), Pro-Val-Leu-Asp-Glu-Phe-Arg-Glu-Lys-Leu-Asn-Glu-Glu-Leu-Glu-Ala-Leu-Lys-Gln-Lys-Leu-Lys (SEQ ID NO:1). Fewer amino acid residues leave a gap in the wheel; more amino acid residues are the reason that certain provisions in this wheel shall be occupied by more than one amino acid residue.

In the case of peptides with 15 or more amino acid residues, such as mimetic ApoA-I containing from 15 to 29 residues, continuous stretch of hydrophobic amino acid residues means that at least one amino acid residue in positions along such a wheel containing two or more amino acid residues, is a hydrophobic amino acid residue. Thus, with reference to figure 2, pho-angle is the arc covered by the remnants of 16, 2, 6, 17, 10, 3 and 14, despite the presence of a hydrophilic residue at position 20, because the residue in position 2, which occupies the same position on the wheel is a hydrophobic residue. Usually mimic ApoA-I with pho-angle in the range from 160°up to 220°, are considered to be entering them in the scope of the present invention. In certain embodiments pho-angle enclosed in the region of 180°up to 200°.

In the peptide 16 (Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:16), or its pharmaceutically acceptable salt, a visual mimetic ApoA-I, positively charged amino acid residues clustered at the last N-terminal turn of the spiral. Although the inventors do not wish to bind themselves to any particular theory, however, they suggest that the cluster of basic residues at the N-end stabilizes the helix in the electrostatic interactions of the dipole charge (NH3+-the spiral. It is also understood that stabilization occurs as a result of hydrophobic interactions between the side chains of lysine and the Central part (core) of the spiral (see, Groebke et al., 1996, Proc. Natl. Acad. Sci. U.S.A. 93: 4025-4029; Esposito et al., 1997, Biopolymers 41: 27-35).

With the exception of a positively charged N-terminal cluster of negative charges in the peptide 16 or its pharmaceutically acceptable salt are located in the remaining part of the hydrophilic surface of at least one negatively charged (acidic) amino acid residue on the coil, resulting in a continuous period of negative charges along the hydrophilic surface of the spiral. One positive charge is localized on the rest 16, which potentially contributes to stabil the activity spiral by formation of a salt bridge with an acidic residue on the adjacent turn of the spiral.

It is believed that using NMR studies of peptide 16 or its pharmaceutically acceptable salt could demonstrate that amino acid residues 13, 14, 17 and 20 of the above peptide forming a hydrophobic cluster near the C-Terminus of the helix. Phe 17 is located in the center of this cluster, and are believed to play an important role in the stabilization of this hydrophobic cluster.

Although the inventors do not wish to bind themselves to any particular theory, nevertheless they believe that the hydrophobic cluster formed by residues 13, 14, 17 and 20 of the peptide 16 or its pharmaceutically acceptable salt is essential in the implementation of the lipid binding and activation of LCAT. It is believed that amphiphilic peptides will contact the phospholipids are oriented with their hydrophobic surface in the direction of the alkyl chains of the lipid fragments. Thus, it is believed that the specified vysokopetrovsky cluster contributes to ensuring a high affinity for lipids observed in relation mimetics of ApoA-I according to the invention. Since binding of lipids is a prerequisite for the activation of lecithin-cholesterol-acyltransferase (LCAT), it is believed that the hydrophobic cluster is also important to activate LCAT.

Aromatic residues may be important in zakalivanie peptides and proteins to whether the idam (De Kruijff, 1990, Biosci. Rep. 10: 127-130; O'neil and De Grado, 1990, Science 250: 645-651; Blondelle et al., 1993, Biochim. Biophys. Acta 1202: 331-336). Thus, in addition, it is believed that the Phe-17, which is located in the Central hydrophobic cluster, can also play an important role in zakalivanie peptide 16 or its pharmaceutically acceptable salt to lipids.

The long axis of the α-helix formed by the mimic ApoA-I, in General, has a generally curved configuration. In a typical amphiphilic helix found that the length of the hydrogen bonds hydrophilic and hydrophobic surfaces vary in such a way that the hydrophobic side of the helix is concave (Barlow and Thornton, 1988, J. Mol. Biol. 201: 601-619; Zhou et al., 1992, J. Am. Chem. Soc. 33: 11174-11183; Gesell et al., 1997, J. Biomol. NMR 9: 127-135). Although the inventors do not wish to bind themselves to any particular theory, nevertheless they believe that the total curvature of the hydrophobic surface of the spiral may be important for binding disc-shaped complexes curved spiral allows the peptide to better "match" the edges around the disc-shaped particles, thereby increasing the stability of the complex of the peptide with the disk.

In the conventional structural model ApoA-I amphiphilic α-helix are grouped around the edges discoid HDL (see figv). In this model it is assumed that the spiral is built along their hydrophobic surfaces and directed to the lipid acyl CE and (Brasseur et al., 1990, Biochim. Biophys. Acta 1043: 245-252). Helix are antiparallel manner, and it is assumed that a cooperative effect between the coils contributes to the stability of the disc-shaped complex of HDL (Brasseur et al., above). It has been suggested that one of the factors that contributes to the stability of the disc-shaped complex of HDL, is the existence of the ionic interactions between acidic and basic residues, leading to the formation of intermolecular salt bridges or hydrogen bonds between residues on neighboring antiparallel helix. In this model, the peptides are not treated as a separate object, but in interaction with at least two other adjacent peptide molecules (pigv).

It is generally accepted that the intramolecular hydrogen bond or salt bridge formed between acidic and basic residues, respectively, in positions i and i+3 spiral, stabilizes the helical structure (Marqusee et al:, 1985, Proc. Natl. Acad. Sci. USA 84(24): 8898-8902).

Thus, the mimic ApoA-I possess the ability to form intermolecular hydrogen bonds with each other when they are arranged in antiparallel manner and their hydrophobic surface rotated in the same direction, as in the case when the peptides are associated with lipids. The mimic ApoA-I also have what sposobnostey to form intramolecular hydrogen bonds or salt bridges near the N - and C-ends of the spiral.

Moreover, at the location specified in antiparallel manner spirals are closely Packed; there are no steric constraints that prevent close contact between the spirals. The mimic ApoA-I have the ability to dense packing and ionic interaction, with the formation of intra - and/or intermolecular salt bridges and/or hydrogen bonds with the binding of lipids in antiparallel manner.

The mimic ApoA-I is capable of self. The phenomenon of self depends on the conditions of pH, concentration of peptide and ionic strength, can result in multiple States Association, from Monomeric to several variants of multimeric forms (figa). The hydrophobic core of the peptide aggregates favorable hydrophobic interactions with lipids. The ability of the peptides to aggregate even at very low concentrations may contribute to their binding to lipids. It is assumed that the center of the peptide aggregates arise peptide-peptide interactions, and they can compete with the lipid-peptide interactions.

The hydrophobic core units mimetics of ApoA-I promotes interactions with lipids. The ability mimetics of ApoA-I to aggregation even at very low concentrations may contribute to their binding to lipids. Interaction between mimic ApoA-I elapidae leads to the formation of peptide-lipid complexes. As illustrated in figa received type complex (comically, disks, vesicles or multilayers) may depend on the lipid:peptide molar ratio, with comically usually formed at low lipid:peptide molar relationship, and with the increase of molar relationships are formed disc-shaped and vesicular or multi-layer complexes. Micelles are usually formed approximately at the ratios of 2 mol of lipid approximately 1 mol ApoA-I, or about 2 mol lipid approximately 6-10 mol of mimetica ApoA-I. disc-Shaped complexes are usually formed at the ratios, constituting approximately 50-100 mol lipid: approximately 1 mol ApoA-I, or from about 6 to about 10 mol of mimetica ApoA-I. Vesicular complexes are usually formed at the ratios, components from approximately 200 to approximately 300 mol lipid: approximately 1 mol ApoA-I, or from about 6 to about 10 mol of mimetica ApoA-I. the Specified property was described for amphiphilic peptides (Epand, The Amphipathic Helix, 1993) and ApoA-I (Jones, 1992, Structure and Function of Apolipoproteins, Chapter 8, pp.217-250). Lipid-peptide molar ratio also determines the size and composition of such complexes.

D. Modified forms of the peptides of formulas I, II and III and their pharmaceutically acceptable salts

In other embodiments, the mimetics ApoA-I containing 22 or less amino acid mod is impressive. Of course, truncated or internally shortened form of the formula I, II or III containing 21, 20, 19, 18, 17, 16 or even 15 amino acid residues, which essentially retain the overall characteristics and properties of amphiphilic helices formed mimic ApoA-I, are included in the scope covered by the present invention.

In one of the embodiments of the present invention truncated forms mimetics of ApoA-I is obtained by deletion of one or more amino acid residues from the N - and/or C-Terminus. Shortened due to internal form regions mimetics of ApoA-I is obtained by deletion of one or more amino acid residues occupying positions within mimetics of ApoA-I. Subjected to internal deletions of amino acid residues can be consecutive residues or are not following each other remains.

Professionals in this field should be clear that the internal deletion of amino acid residue in the mimetic ApoA-I can call on the spot deletions rotate the plane of section of hydrophilic-hydrophobic surface of the spiral at 100°. Since such rotation can significantly change amphiphilic properties resulting from this spiral, in one of the embodiments of the present invention one or more amino acid residues subjected to deletions, so that essentially preserved the orientation the plane of the hydrophilic-hydrophobic surface all along the helical axis.

This is conveniently accomplished by deletion of a sufficient number of consecutive or not consecutive amino acid residues, so that the deletion was subjected to one full turn of the spiral. Idealized α-helix is of 3.6 residues on stage. Thus, in one of the embodiments deletions expose groups of 3-4 consecutive or not consecutive amino acid residues. The question of whether to expose the deletion of 3 amino acid residue or 4 amino acid balance, may depend on the position inside the spiral of the first exposed deletions balance. Determining the approximate number of consecutive or not consecutive amino acid residues that constitute one complete turn of the helix from any particular starting point within the amphipatic helix, is fully within the competence of specialists in this field.

The mimic ApoA-I can be extended from one or both ends, or due to internal areas by adding amino acid residues that do not cause harmful interference, and in some embodiments even reinforce structural and/or functional properties of the peptides. Of course, extra long mimetics of ApoA-I containing 23, 24, 25, 26, 27, 28 or 29 amino acid residues, are included in the scope, coverage is offered by the present invention. Such lengthening mimetics of ApoA-I can essentially save total affilinet and other properties of mimetics of ApoA-I. Professionals in this area of bodø, of course, it is clear that the addition of amino acid residues in the inner area can cause the rotation of the plane of the hydrophilic-hydrophobic surfaces on site include an extension residue, similar to that described above for the case of internal deletions. Thus, the considerations discussed above in connection with internal deletions, which also applies to additions of amino acid residues in an inner scope.

In one of the embodiments of the mimic ApoA-I lengthened at their N-and/or C-end, by adding the amino acid sequence containing from 1 to 7 residues.

In one of the embodiments of the mimic ApoA-I lengthened at their N-and/or C-end of at least one turn of the spiral. Such lengthening stabilize the secondary structure of the helix in the presence of lipids, as previously described terminal amino acid residues and segments.

In another embodiment, the mimetics ApoA-I extend to N-end-on-one basic amino acid residue such as Lys (K). In one of the embodiments of the X1represents Lys, X2represents Lys, X3represents Leu, X4represents Lys, X5represents Gln, X6represents Lys, X7p is ecstasy a Leu, X8represents Ala, X9represents Glu, X10represents Leu, X11represents Leu, X12represents Glu, X13represents Asn, X14represents Leu, X15represents Leu, X16represents Glu, X17represents Arg, X18represents Phe, X19represents Leu, X20represents Asp, X21represents Leu, X22represents Val and X23represents Inp.

In the scope of the present invention also includes a "protected" form mimetics of ApoA-I, i.e. forms mimetics of ApoA-I in which R1represents aminosidine group and/or R2represents a carboxyl protective group. It is believed that the removal of N - and/or C-terminal charges mimetics of ApoA-I containing 18 or less amino acid residues (through synthesizing N-acylated peptide amides/complex ether/hydrazides/alcohols and their deputies) can lead to the formation of mimetics that are approaching, and in some embodiments even surpass, the activity unprotected forms such mimetica. In certain embodiments, when such mimetics containing 22 or more amino acid residues, it is believed that blocking the N - or C-Terminus can lead to the formation of mimetics of ApoA-I, kotoryeprodayut lower activity, than non-blocked form. However, the protection of both N - and C-ends, mimetics of ApoA-I containing 22 or more amino acid residues, can restore activity. Thus, in one of the embodiments of the present invention is protected or N-and/or C-end (in another embodiment each end) mimetics of ApoA-I containing 18 or less amino acid residues, whereas the N - and C-ends of the peptides containing 22 or more amino acid residues are either both protected, or both unprotected. Usually N-terminal blocking groups include RC(O)-, where R represents-H, (C1-C6)-alkyl, (C2-C6)-alkenyl, (C2-C6)-quinil, (C5-C20)-aryl, (C6-C26-aralkyl, 5-20-membered heteroaryl or 6-26 membered algeterror. Specific N-terminal blocking groups include acetyl, formyl, dansyl.

Typical C-terminal blocking groups include-C(O)NRR and-C(O)OR, where each R is independently as defined above. Specific C-terminal blocking groups include those in which each R is independently stands. Though not wishing to be bound to any specific theory, the inventors nevertheless believe that such end-blocking groups stabilize the α-helix in the presence of lipids (see, for example, Venkatachelapathi et al., 1993, PROTEINS: Structure, Function, Genetics nd 15: 349-359).

E. Dimers, trimers, tetramer and multimeric peptides of formula I, II or III and their pharmaceutically acceptable salts

The structure of native ApoA-I contains seven helical elements that, considered together, are involved in the binding of lipids (Nakagawa et al., 1985, J. Am. Chem. Soc. 107: 7087-7092; Anantharamaiah et al., 1985, J. Biol. Chem. 260: 10248-10262; Vanloo et al., 1991, J. Lipid Res. 32: 1253-1264; Mendez et al., 1994, J. Clin. Invest. 94: 1698-1705; Palgunari et al., 1996, Arterioscler. Thromb. Vase. Biol. 16: 328-338; Demoor et al., 1996, Eur. J. Biochem. 239:74-84). Thus, the present invention also includes dimers, trimers, tetramer and even polymers of higher order ("multimer") mimetics of ApoA-I. Such multimer can be represented in the form of tandem repeats, branched chains, or combinations thereof. The mimic ApoA-I can be connected directly to each other or separated by one or more linkers.

The mimic ApoA-I, which contain multimer can be peptides of formula I, II or III, analogs of formulas I, II or III, modified forms of the formula I, II or III, truncated forms, or forms with internal deletions of the formula I, II or III, elongated forms of the formula I, II or III and/or their combinations. The mimic ApoA-I can be connected on the principle of head-to-tail (i.e., N-end C-end), on the principle of head-to-head (i.e., N-end N-end), tail-to-tail (i.e. From the end to the C-end), or in various combination is in the form of their pharmaceutically acceptable salts.

In one of the embodiments of the present invention multimer are tandem repeats of two, three, four and approximately ten mimetics of ApoA-I. In one of the embodiments of multimer are tandem repeats from 2 to 8 peptides. Thus, in one of the embodiments of the present invention is connected with multimarine having the following structural formula:

in which

each m independently is an integer from 0 to 1, and in one of the embodiments m is 1; n is an integer from 0 to 10, and in one of the embodiments, n is an integer from 0 to 8; each "HH" independently is a radical derived from mimetica ApoA-I; and each "LL" independently represents a linker.

In structure (IV) a linker LL may be any bifunctional molecule capable of covalent binding of the two peptides with each other. Thus, suitable linkers are bifunctional molecules, in which a functional group capable of covalently joining the N-and/or C-Termini of the peptide. Functional groups suitable for attachment to the N - or C-Termini of peptides is well known to specialists in this field and have a great chemistry to provide education such covalent bonds.

The linker may be flexible, rigid or semi-rigid, depending on the desired properties of multimer. the right linkers include, for example, amino acid residues such as Pro, azPro, Pip, azPip or Gly, or peptide segments containing from about 2 to about 5, 10, 15 or 20 or even more amino acid residue of a bifunctional organic compounds, such as H2N(CH2)nCOOH, HO(CH2)nCOOH and HO(CH2CH2O)nCH2CH2COOH, where n is an integer from 1 to 12, etc. are Examples of such linkers, as well as methods of obtaining such linkers and compounds, including such linkers are well known in the art (see, for example, Hunig et al., 1974, Chem. Ber. 100: 3039-3044; Basak et al., 1994, Bioconjug. Chem. 5(4): 301 to 305).

In one of the embodiments of the present invention tandem repeats internally interrupted only prolinnova balance. In those cases, when the mimic ApoA-I does not contain N - or C-terminal polynomy balance, LL can be Pro, D-Pro, azPro, Pip, D-Pip or azPip, and m is 1.

In certain embodiments of the present invention may be necessary the use of fissionable linkers that provide a release of one or more helical segments (HH) under certain conditions. Suitable degradable linkers include peptides having the sequence of amino acid residues that are recognized by proteases, oligonucleotides, which can be cleaved by endonucleases, and organic link is, which can be broken down by chemical means, for example, in acidic, alkaline or other conditions. Typically, the cleavage conditions are relatively mild, so as not to cause denaturation or to otherwise cause degradation of the helical and/or unsplit of linkers that form multimeric.

Peptide and oligonucleotide linkers that can be selectively split, as well as ways of splitting these linkers are well known in this area and should be obvious to specialists. Suitable linkers in the form of organic compounds that can selectively to split, well known to specialists in this field and include those described, for example, in the application WO 94/08051, and quoted in her publications.

In one of the embodiments is used by linkers include peptides that are substrates for endogenous circulating enzymes, thus providing the opportunity to multimers selectively splitin vivo. Endogenous enzyme suitable for cleavage of the linkers, for example, is Pro-apolipoprotein A-I-propeptides. The enzymes, as well as peptide segments that act as substrates for such enzymes are well known in the art (see, for example, Edelstein et al., 1983, J. Biol. Chem. 258: 11430-1143; Zanis, 1983, Proc. Natl. Acad. Sci. USA 80: 2574-2578).

In one of the embodiments use linkers satisfactory length and flexibility, in order to provide antiparallel orientation of spiral segments (HH) (II) and the formation of intermolecular hydrogen bonds or salt bridges in the presence of lipids. Linkers satisfactory length and flexibility include, but are not limited to, the residue or radical Pro, D-Pro, azPro, Pip, D-Pip, azPip, Gly, Cys-Cys, H2N(CH2)nCOOH, HO(CH2)nCOOH or HO(CH2CH2O)nCH2CH2COOH, where n takes values from 1 to 12, or from 4 to 6; H2N-aryl-COOH and hydrocarbons.

Alternatively, since native apolipoprotein provide the possibility of cooperative binding between antiparallel helical segments, peptide linkers, which primary sequence correspond to peptide segments connecting adjacent spiral native apolipoproteins, including, for example, ApoA-I, ApoA-II, ApoA-IV, ApoC-I, ApoC-II, ApoC-III, ApoD, ApoE and ApoJ, can be suitably used for binding mimetics of ApoA-I formula I. Such sequences are well known in the art (see, for example, Rosseneu et al., "Analysis of the Primary and of the Secondary Structure of the Apolipoproteins," In: Structure and Function of Lipoproteins, Ch.6, 159-183, CRC Press, Inc., 1992).

Other linkers that provide education mimola olyarnik hydrogen bonds or salt bridges between tandem repeats antiparallel helical segments, include peptides with turns in the opposite direction, such as β-turns and γ-turns, as well as organic molecules that mimic the structure of the peptide β-bends and/or γ-curves. Usually turns in the opposite direction are segments of the peptide, which change the direction of the polypeptide chain on the opposite, to create a separate polypeptide chains to associate with areas of antiparallel β-pleated structure or antiparallel α-helical structure. β-curves are usually composed of four amino acid residues, and γ-curves are usually composed of three amino acid residues.

Conformation and sequence of many peptide β-curves are described in detail in the field and include, for example, not in the order constraints, type-I, type-I, type-II, type-II, type-III, type-III', type IV, type V, type-V',-type VIa type VIb, type VII and type VIII (see Richardson, 1981, Adv. Protein Chem. 34: 167-339; Rose et al., 1985, Adv. Protein Chem. 37: 1-109; Wilmot et al., 1988, J. Mol. Biol. 203: 221-232; Sibanda et al., 1989, J. Mol. Biol. 206: 759-777; Tramontano et al., 1989, Proteins: Struct. Funct. Genet. 6: 382-394).

The specific conformation of short peptide curves, such as β-turns, depends primarily on the provisions of certain amino acid residues in bending (usually Gly, Asn, or Pro). Generally, β-bend type-I is compatible with any amino acid residue positions 1 to 4 of this curve, with the only exception is receiving, that Pro cannot be in position 3. Gly prevails in position 4 and Pro prevails in position 2 of the curves of both types, type-I and type-II. Residues Asp, Asn, Ser and Cys often found in position 1, where their side chains often form a hydrogen bond with the NH of residue 3.

The curves of type-II residues Gly and Asn are found most often in position 3, because they are most easily accept the corners of the frame. In the ideal case, the curves of type-I' is Gly at positions 2 and 3, and in the bends of the type-II' is Gly in position 2. Curves of type-III can usually have the greatest number of amino acid residues, but the curves of type-III', as a rule, require a Gly at positions 2 and 3. Curves of type-VIa and VIb usually contain a CIS-peptide bond and Pro as an internal balance. As an overview of different types and sequences of β-bends in proteins and peptides, the reader should refer to the publication Wilmot et al., 1988, J. Mol. Biol. 203: 221-232.

Conformation and sequence many pipinich γ-curves are also well documented in this area (see, for example, Rose et al., 1985, Adv. Protein Chem. 37: 1-109; Wilmer-White et al., 1987, Trends Biochem. Sci. 12: 189-192; Wilmot et al., 1988, J. Mol. Biol. 203: 221-232; Sibanda et al., 1989, J. Mol. Biol. 206: 759-777; Tramontano et al., 1989, Proteins: Struct. Funct. Genet. 6: 382-394). All of these types of structures of β-turns and γ-curves and their corresponding sequences, as well as open later patterns and sequence p is tidnyj β-turns and γ-turns, specifically included in this description.

Alternatively, the linker (LL) may contain organic molecule or fragment that mimics the structure of the peptide β-bend or γ-curve. These mimic the β-bend and/or γ-bending fragments, as well as methods of synthesizing peptides containing such fragments are well known in this field and include, without limitation, those described in Giannis and Kolter, 1993 Angew. Chem. Intl. Ed. Eng. 32: 1244-1267; Kahn et al., 1988, J. Molecular Recognition 1: 75-79; Kahn et al., 1987, Tetrahedron Lett. 28: 1623-1626.

In another of the embodiments of the present invention multimer presented in the form of branched chains (see, e.g., figure 3). Such networks conveniently be obtained by using multi-functional binding fragments, which provide connection to a simple binding fragment more than two helical elements. Thus, in branched networks are molecules that have three, four or more functional groups capable of covalently to contact the N - and/or C-end of the peptide. Suitable binding fragments include, for example, amino acid residues having side chains with functional groups hydroxyl, sulfonyl, amino, carboxyl, amide and/or ether complex, such as Ser (S)Thr (T), Cys (C), Tyr (Y), Asn (N), Gln (Q), Lys (K), Arg (R), Orn, Asp (D) and Glu (E); and the corresponding D-Aisne is timer each of the above; or the remains of other organic molecules containing these functional groups.

Helical segments that are connected to a separate connecting piece does not need to be attached to the same ends. In fact, in certain embodiments the spiral segments attached to a separate connecting piece in such a way that they are arranged in antiparallel structure, i.e. that some of the coils were attached via its N-ends, and the rest through its C-ends.

Spiral segments can be attached directly to the connecting piece or, as described earlier, can be spatially separated from the binding of a fragment of one or more bifunctional linkers (LL).

If we turn to figa and 3B, one can see that the branched network can be described in terms of the number of nodes forming the network, where each multi-binding fragment forms a node. On figa and 3B helical segments (i.e. mimic ApoA-I) is illustrated in the form of a cylinder, and a multifunctional binding fragments (or nodes) in the form of circles (●), where the number of lines coming out of the circle indicates the "order" (or the number of functional groups) multifunctional binding fragment.

The number of nodes in the network will typically depend on the total Tr the required number of spiral segments, and as a rule, will be approximately from 1 to 2. Of course, it would be desirable to ensure that the required number of spiral segments, the network having binding fragments of higher order, would have fewer nodes. For example, if you refer to figa and 3B, the network of the third order (i.e. a network that has trifunctionally binding fragments) of the seven helical element has three nodes (figa), whereas the network of the fourth order (that is, a network that has tetrafunctional binding fragments) of the seven helical elements has only two nodes (pigv).

The network can be uniform (the same) order, that is, they can be a network in which all nodes are, for example, trifunctional or tetrafunctional binding fragments, or may be mixed networks, such as networks, in which nodes are mixed nodes, as, for example, trifunctional or tetrafunctional binding fragments. Of course, it should be understood that even in the case of networks of uniform order is not necessary that the connecting pieces were identical. In networks of the third order can be used, for example, two, three, four or even more different trifunctionally binding fragments.

Like linear multimers, spiral elements, containing an extensive network, can is to be but not necessarily, identical.

An example of such an extensive network of mixed order is illustrated in figs. On figs helical segments (i.e. mimic ApoA-I) is illustrated in the form of a cylinder, and a multifunctional binding fragments in the form of circles (●), where the number of lines coming out of the circle indicates the "order" (or the number of functional groups) multifunctional binding fragment. The line connecting the spiral segments are as described above, bifunctional linkers LL. Helical segments that are contained in branched networks, can, as described above, to be tandem repeats mimetics of ApoA-I.

In one of the illustrative embodiments extensive network according to the invention are described by the following formula:

in which

each X independently is a radical derived from multimer the following formula:

in which

each HH is independently a radical derived from mimetica ApoA-I;

each LL is a bifunctional linker;

each m independently is an integer from 0 to 1;

each n independently is an integer from 0 to 8;

each of the Nyaand Nybindependently is a multifunctional binding fragment, where where yaand ybrepresent the number of functional groups accordingly, the Nyaand Nyb; each yaand ybindependently is an integer from 3 to 8; and p is an integer from 0 to 7.

In one of the embodiments of the extensive network contains Lys tree" ("lysine tree"), that is, a network in which a multifunctional binding fragment represents one or more residues Lys (K) (see, for example, fig.3D).

In one of the illustrative embodiments extensive network specified "lysine tree" ("Lys tree") according to the invention as described by the following formulas:

in which

each X independently is a radical derived from multimer the following formula:

in which

each HH is independently a radical derived from mimetica ApoA-I, formula I;

each LL is a bifunctional linker;

each n independently is an integer from 0 to 8;

each m independently is an integer from 0 to 1;

R1represents an-OR or-NRR; and

each R independently represents-H, (C1-C6)-alkyl, (C2-C6)-alkenyl, (C2-C6)-quinil or (C5-C20)-aryl.

Some additional illustrative mimic ApoA-I are presented below in table 7:

or their pharmaceutically acceptable salt.

Some illustrative mimic ApoA-I with acetylated N-end and aminirovanie C-end, is presented below in tables 8 and 9:

or their pharmaceutically acceptable salt.

or their pharmaceutically acceptable salt.

III. Synthesis of mimetics of ApoA-I

The mimic ApoA-I may actually be obtained using any known in the field of technology for peptides. For example, mimetics ApoA-I can be obtained by standard methods manual peptide synthesis in solution or solid-phase peptide synthesis, or by using recombinant DNA technologies.

i> A. Chemical synthesis

The mimic ApoA-I can be obtained using standard methods manual peptide synthesis in the liquid phase or solid-phase peptide synthesis (see, e.g., Chemical Approaches to the Synthesis of Peptides and Proteins, Williams et al., Eds., 1997, CRC Press, Boca Raton Fla., and are there links; Solid Phase Peptide Synthesis: A Practical Approach, Atherton &Sheppard, Eds., 1989, IRL Press, Oxford, England, and are there links).

Alternatively, the mimic ApoA-I can be obtained by segment condensation of peptides, as described, for example, Liu et al., 1996, Tetrahedron Lett. 37(7): 933-936; Baca, et al., 1995, J. Am. Chem. Soc. 117: 1881-1887; Tarn et al., 1995, Int. J. Peptide Protein Res. 45: 209-216; Schnolzer and Kent, 1992, Science 256: 221-225; Liu and Tarn, 1994, J. Am. Chem. Soc. 116(10): 4149-4153; Liu and Tarn, 1994, Proc. Natl. Acad. Sci. USA 91: 6584-6588; Yamashiro and Li, 1988, Int. J. Peptide Protein Res. 31: 322-334). It is appropriate, in particular, in the case of peptides with a glycine residue. Other methods used for the synthesis of mimetics of ApoA-I, are described in Nakagawa et al., 1985, J. Am. Chem. Soc. 107: 7087-7092.

The mimic ApoA-I with N - and/or C-terminal blocking groups can be obtained using standard technologies of organic chemistry. For example, methods of acylation of N-Terminus of the peptide or amidation or esterification of the C-terminal end of the peptide are well known in this field. The ways of making other modifications to the N - and/or C-end known to the specialists in this field, as well as ways to protect any functional the groups of the side chains, may be necessary to attach a terminal blocking groups.

Pharmaceutically acceptable salts (counterions) can be obtained by standard methods by ion-exchange chromatography or other well-known in the field of ways.

The mimic ApoA-I, which are presented in the form of a tandem of multimers, can be synthesized by standard methods by adding the linker (linkers) to the peptide chain at an appropriate stage of the synthesis. Alternatively, it is possible to synthesize spiral segments, and each segment can enter into reaction with the specified linker. Of course, the specific method of synthesis will depend on the composition of the linker. Proper protection and the appropriate chemistry is well known and will be obvious to experts in this field.

The mimic ApoA-I, which are presented in the form of branched chains may be synthesized by standard methods using the trimeric and tetramer resins and chemical technologies described Tarn, 1988, Proc. Natl. Acad. Sci. USA 85: 5409-5413 and Demoor et al., 1996, Eur. J. Biochem. 239: 74-84. Modified synthetic resins and strategies for the synthesis of branched networks of higher or lower orders, or those that contain a combination of different helical mimetics of ApoA-I, are within the competence of specialists in the field of PE is tidey chemistry and/or organic chemistry.

Education, if necessary, the disulfide bond can be carried out in the presence of mild oxidizing means. Can be used chemical oxidizing means, or mimetics ApoA-I may just be exposed to air to form such relationships. In this area known various methods, including the methods described, for example, Tarn et al., 1979, Synthesis 955-957; Stewart et al., 1984, Solid Phase Peptide Synthesis, 2d Ed., Pierce Chemical Company, Rockford, 111; Ahmed et al., 1975, J. Biol. Chem. 250: 8477-8482; and Pennington et al., 1991 Peptides 1990, 164-166, Giralt and Andreu, Eds., ESCOM Leiden, The Netherlands. An additional alternative is described Kamber et al., 1980, Helv. Chim. Acta 63: 899-915. A method conducted on solid supports is described by Albericio, 1985, Int. J. Peptide Protein Res. 26: 92-97. Any of these methods can be used for the formation of disulfide bonds in peptides according to the invention.

The mimic ApoA-I containing one or more internal glycine residues, can be synthesized with a relatively high yield by segment condensation of peptides, thus providing advantages for large-scale production. Segment condensation, i.e. the Union of small chains of the constituent peptides with the formation of a larger peptide chain is used for production of many biologically active peptides, including mimetics ApoA-I, with 44 amino acid residue (see, for example, Nakagawa et al., 1985, J. Am Chem. Soc. 107: 7087-7083; Nokihara et al., 1989, Peptides 1988: 166-168; Kneib-Cordonnier et al., 1990, Int. J. Pept. Protein Res. 35: 527-538).

Advantages of synthesis through the segment condensation include the ability to condense the pre-formed segments in the liquid phase and the ease of purification of the final product. The disadvantages of this method include low levels of efficiency of cooperation and exit at the stage of condensation, as well as the low solubility of certain peptide sequences. The effectiveness of the interaction at the stage of condensation can be increased by increasing the interaction time. Usually the longer interaction leads to increased racemization of the product (Sieber et al., 1970, Helv. Chim. Acta 53: 2135-2150). However, since glycine is deprived chiral center, it does not undergo racemization (prolinnova remains, due to steric hindrances, also a little prone or not prone to racemization during the long time of interaction). Thus, embodiments of which are associated with the presence of internal glycine residues, the synthesis through a radial condensation can be carried out in large quantities and with great output by synthesizing the electoral segments, which can be an advantage due to the fact that glycine residues do not undergo racemization. Thus the om, the mimic ApoA-I containing one or more internal glycine residues provide the advantage in the synthesis due to the possibility of obtaining large amounts of the drug.

B. Recombinant synthesis

If mimetic ApoA-I consists entirely of genetically encoded amino acid residues, or if part of it is from them, then such mimetic ApoA-I or the relevant part thereof can also be synthesized using standard recombinant technology of genetic engineering.

To obtain recombinant products polynucleotide sequence encoding the corresponding peptide, was built in suitable expressing vector, i.e. a vector which contains the necessary elements for the transcription and translation of the built-in coding sequence, or, in the case of vector RNA virus, the necessary elements for replication and translation. Then expressing the media transferout into a suitable host cell which will Express the indicated peptide. Then, depending on how expressing system is used, expressed peptide allocate one or another of the well-known in the field of methods. Methods of producing recombinant proteins and peptides are well known in the art (see, for example, the publication of Sambrook et al., 1989, Molecular Cloning A Laboatory Manual, Cold Spring Harbor Laboratory, N. Y.; and Ausubel et al., 1989, Current Protocols in Molecular Biology, Greene Publishing Associates and Wiley Interscience, N. Y., each of which is fully incorporated into this description by reference).

To improve the efficiency of production polynucleotide can be designed in such a way that it encodes many elements peptide, separated by areas of enzymatic degradation of either of the homopolymers (repeating peptide elements), or heteropolymers (various peptides that are attached to each other). The resulting polypeptide can be cleaved (e.g., by treatment with an appropriate enzyme) in order to restore peptide elements. This can improve the yield of peptides under the control of a single promoter. In one of the embodiments of the polycistronic polynucleotide can be designed in such a way as to transcarbamoylase a single mRNA, which encodes multiple peptides (i.e. homopolymers or heteropolymers), while to each of the coding region was operatively associated with the cap-independent sequence exercising control broadcast; for example, with a plot of the internal landing ribosomes (IRES). When used in appropriate virus expressing systems broadcast each peptide encoded by the mRNA, is directed inwardly in the transcript, such as the er, using the IRES. Thus, a polycistronic structure directs the transcription of a single large polycistronic mRNA, which, in turn, forwards the broadcast of multiple individual peptides. This approach eliminates the production and enzymatic processing of polyproteins and can significantly increase the yield of the peptide under the control of a single promoter.

For the expression of mimetics of ApoA-I can be used in different vector expressing the system host cells. Such systems include, but are not limited to, microorganisms such as bacteria transformed with recombinant DNA bacteriophage or plasmid DNA containing an appropriate coding sequence; yeast or filamentous fungi transformed with recombinant yeast or fungal expressing vectors containing an appropriate coding sequence; insect cells infected with vectors expressing recombinant virus (e.g. baculovirus)containing an appropriate coding sequence; plant cell infected with vectors expressing recombinant virus (e.g., cauliflower mosaic virus)or transformed with vectors expressing recombinant plasmid (in the example, Ti-plasmid)containing an appropriate coding sequence; or animal cells.

The elements of expression expressing systems can vary in efficiency and specificlaly. Depending on the system host/vector, any number of suitable elements for transcription and translation, including constitutive and inducible promoters, may be used in expressing vector. For example, when cloning in bacterial systems can be used inducible promoters such as pL of bacteriophage λ, plac, ptrp, ptac (hybrid promoter ptrp-lac) and the like; when cloning in insect cells can be used such as promoters baculovirus polyadrny promoter; when cloning in plant cells can be used promoters derived from the genome of plant cells (e.g., promoters [proteins] heat shock; the promoter of the small subunit of RUBISCO; the promoter a/b binding protein chlorophyll) or from plant viruses (e.g., the 35S promoter-RNA virus, CaMV; the promoter of the protein shell TMV); when cloning in mammalian cells can be used promoters derived from the genome of mammalian cells (for example, the promoter metallothionein) or from mammalian viruses (e.g., the late promoter of adenovirus; the promoter of the virus cows is it virus 7.5 so-called); when generating cell lines that contain multiple copies of the expressed product may be used, vectors based on SV40, BPV-and EBV with an appropriate selective marker.

When used expressing vectors of plants, the expression of sequences encoding the mimic ApoA-I can be running any of a number of promoters. For example, can be used viral promoters such as the 35S promoter-RNA and 19S-RNA virus CaMV (Brisson et al., 1984, Nature 310: 511-514), or the promoter of the protein shell TMV (Takamatsu et al., 1987, EMBO J. 6: 307-311); alternatively, it may be used promoters plants, such as the promoter of the small subunit of RUBISCO (Coruzzi et al., 1984, EMBO J. 3: 1671-1680; Broglie et al., 1984, Science 224: 838-843) or promoters [proteins] heat shock, for example, hspl7.5-E or hspl7.3-B soybean (Gurley et al., 1986, Mol. Cell. Biol. 6: 559-565). These designs can be integrated into plant cells using Ti plasmids, Ri plasmids, vectors of plant viruses by direct DNA transformation, microinjection, electroporation, etc. as an overview of these technologies, see, for example, Weissbach &Weissbach, 1988, Methods for Plant Molecular Biology, Academic Press, N. Y., Section VIII, pp.421-463; and Grierson &Corey, 1988, Plant Molecular Biology, 2d Ed., Blackie, London, Ch.7-9.

In one of expressing systems of insects, which can be used for producing mimetics of ApoA-I, Autographa californica virus cores is wow polyhedrosis (AcNPV) is used as the vector for expression of foreign genes. The virus grows in the cells of Spodoptera frugiperda. The coding sequence can be cloned in the field of non-vital (for example, gene politana) regions of the virus and placed under control of an AcNPV promoter (for example, policronio promoter). Successful embedding the coding sequence will result in inactivation of the gene politana and products noccluderindex recombinant virus (i.e., a virus lacking the protein shell encoded poliekrannym genome). Then these recombinant viruses are used to infect cells of Spodoptera frugiperda, in which the integrated gene is expressed (see, for example, Smith et al., 1983, J. Virol. 46: 584; Smith, U.S. patent No. 4215051). Additional examples of such expression systems can be found in Current Protocols in Molecular Biology, Vol.2, Ausubel et al., eds., Greene Publish. Assoc. & Wiley Interscience.

In host mammalian cells can be used a number expressing the systems on the basis of the virus. In cases where as expressing vector is used, the coding sequence can be Legerova with complex, which control the transcription/translation of adenovirus, for example, with the sequence of the late promoter and three of the leader sequence. Then this chimeric gene may be integrated into an adenoviral genome by recombinationin vitr orin vivo. In the installation in an area which is a vital region of the viral genome (e.g., region El or E3)will be obtained recombinant virus that is viable and will be able to ekspressirovali peptide in inzerovanych host cells (see, e.g., Logan &Shenk, 1984, Proc. Natl. Acad. Sci. (USA) 81: 3655-3659). Alternatively, it may be used by the promoter vaccines 7.5 K (see, for example, Mackett et al., 1982, Proc. Natl. Acad. Sci. (USA) 79: 7415-7419; Mackett et al., 1984, J. Virol. 49: 857-864; Panicali et al., 1982, Proc. Natl. Acad. Sci. 79: 4927-4931).

Other expressing system for producing mimetics of ApoA-I will be obvious to specialists in this field.

C. Clean

The mimic ApoA-I can be purified using known in the field of technologies such as chromatography with reversed-phase high-performance liquid chromatography, ion exchange chromatography, gel electrophoresis, affinity chromatography and the like, the Specific conditions used for the purification of specific mimetica ApoA-I, may partly depend on the strategy of synthesis and factors such as the total charge, hydrophobicity, hydrophilicity, etc. and also will be obvious to specialists in this field. Multimeric branched peptides can be purified, for example by ion-exchange chromatography or exclusive chromatography.

For cleaning by means of the PTO affinity chromatography can be used any antibody, which specifically binds to a mimetic ApoA-I. For the production of antibody is possible to immunize various host animals including, but not limited to, rabbits, mice, rats, etc. by injection of the peptide. The peptide may be linked to an appropriate carrier, such as BSA, through a functional group of a side chain or through linkers attached to the functional group of the side chain. To enhance the immunological response can be used in a variety of adjuvants, depending on the type of owner, including, but not limited to, beta-blockers (complete and incomplete), mineral gels such as aluminum hydroxide, surface active substances such as lysolecithin, surface-active polyols, polyanion, peptides, oil emulsions, hemocyanin lymph snails, dinitrophenol, and potentially useful human adjuvants such as BCG (Bacillus Calmette-Guerin) and Corynebacterium parvum.

Monoclonal antibodies against mimetica ApoA-I can be obtained using any technology that enables the production of antibody molecules stable cell lines in culture. Such technologies include, but are not limited to, the hybridoma technique described by Kohler and Milstein, 1975, Nature 256: 495-497, or Kaprowski, U.S. patent No. 4376110, which is included in the present description in widescale; technology human B-cell hybridomas) Kosbor et al., 1983, Immunology Today 4: 72; Cote et al., 1983, Proc. Natl. Acad. Sci. U.S.A. 80: 2026-2030); and technology EBV-hybrid (Cole et al., 1985, Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc., pp.77-96 (1985)). In addition, can be obtained used the technology developed for the production of "chimeric antibodies" (Morrison et al., 1984, Proc. Natl. Acad. Sci. U.S.A. 81: 6851-6855; Neuberger et al., 1984, Nature 312: 604-608; Takeda et al., 1985, Nature 314: 452-454, Boss, U.S. patent No. 4816397; Cabilly, U.S. patent No. 4816567, which is incorporated into this description by reference) by splicing the genes of a mouse antibody molecule with appropriate antigen specificity of the genes for molecules of human antibodies with the appropriate biological activity. Or can be obtained "humanized" antibodies (see, e.g., Queen, U.S. patent No. 5585089, which is included in the present description by reference). Alternatively, techniques described for the production of single-chain antibodies (U.S. patent No. 4946778), can be adapted for producing a peptide-specific single-chain antibodies.

Antibody fragments that contain deletions of specific binding sites, can be obtained using known technologies. For example, such fragments include, but are not limited to F(ab')2-fragments, which can be obtained by treatment with pepsin molecules, antibodies and Fab fragments, which m which may be obtained by restoring the disulfide bridges of F(ab') 2-fragments. Alternatively, it may be constructed expressing Fab libraries (Huse et al., 1989, Science 246: 1275-1281), allowing quick and easy identification of monoclonal Fab fragments with the desired specificity interest peptide.

The antibody or antibody fragment, specific for the desired mimetica ApoA-I, can be attached, for example, agarose, and a complex of the antibody-agarose is used in chromatography for the purification of peptides according to the invention. Cm. Scopes, 1984, Protein Purification: Principles and Practice, Springer-Verlag New York, Inc., N. Y., Livingstone, 1974, Methods In Enzymology: Immunoaffinity Chromatography of Proteins 34: 723-731.

IV. Composition

In one of the embodiments of the present invention relates to compositions containing an effective amount of mimetica ApoA-I and a pharmaceutically acceptable carrier or excipient.

The compositions can be formulated for administration to a mammal by injection. Injectable preparations include sterile suspensions, solutions or emulsions of the active ingredient in aqueous or oily media. The composition can also include prescription agents, such as suspendisse, stabilizing and/or dispersing agents. Compositions for injection may be presented in unit dosage form, e.g., in ampoules or in containers for multiple doses, and could the t contain added preservatives. Alternatively, a composition for injection can be presented in the form of a powder, providing before applying the restoration of a suitable carrier, including, but not limited to, sterile pyrogen-free water, buffer, dextrose, etc. For such purposes mimetic ApoA-I may be dried, or can be obtained with lyophilized peptide-lipid complex. Designed for storage of drugs can be supplied in unit dosage forms and recover before usingin vivo.

For prolonged delivery of specified composition can be prepared in the form of the drug prolonged action for administration by implantation; for example, by subcutaneous, intradermal or intramuscular injection. Thus, for example, mimetic ApoA-I can be prepared with the appropriate polymeric or hydrophobic materials (for example, as an emulsion in an acceptable oil) or ion exchange resins, or as slowly soluble derivatives; for example, in the form of a slowly soluble salt forms of mimetica ApoA-I.

In another embodiment, the composition is injected. Alternatively, it may be used transdermal system for the delivery of percutaneous absorption, manufactured as an adhesive disc or patch which slowly released active and gradient. This purpose can be used amplifiers permeability to facilitate transdermal penetration of mimetica ApoA-I. beneficial effects can be achieved, in particular, by incorporating mimetica ApoA-I in nitroglycerine patch to apply in relation to a mammal having such a Condition as coronary heart disease or hypercholesterolemia.

For oral administration the compositions may be presented, for example, in the form of tablets or capsules, obtained in a standard way in the composition with pharmaceutically acceptable excipients such as binders (e.g., pregelatinized maize starch, polyvinylpyrrolidone or hypromellose); fillers (e.g. lactose, microcrystalline cellulose or dicalcium phosphate); lubricants (e.g. magnesium stearate, talc or silica); disintegrating agents (e.g., potato starch or sodium starch glycolate); or moisturizers (for example, sodium lauryl sulphate). Tablets can be coated by methods well known in the field. Liquid preparations for oral administration may be presented, for example, in the form of solutions, syrups or suspensions, or they may be presented as a dry product for its preparation with water or another suitable carrier is before applying. Such liquid preparations can be obtained by standard means with pharmaceutically acceptable additives such as suspendresume means (for example, sorbitol syrup, cellulose derivatives or hydrogenated edible fats); emulsifying means (e.g., lecithin or acacia); non-aqueous vehicles (e.g. almond oil, oily esters, ethyl alcohol or fractionated vegetable oils); and preservatives (e.g. methyl or propyl-para-hydroxybenzoate or sorbic acid). These preparations may, if desired, also contain buffer salts, flavoring agents, colorants and sweeteners. Preparations for oral administration can be formulated in such a way that it is provided controlled release of mimetica ApoA-I.

For buccal injection composition can be presented in the form of tablets or candies made in the standard way. For rectal and vaginal routes of administration, the active ingredient may be formulated as solutions (for retention enemas)suppositories or ointments.

For administration by inhalation mimetic ApoA-I conveniently delivered in the form of an aerosol spray from pressurized packs or a nebulizer, with the use of a suitable propellant, e.g. DICHLORODIFLUOROMETHANE, trichloromethane, dichlorotetrafluoroethane, and carbon dioxide and other suitable gas. In the case of a pressurized aerosol dosage unit can determine if to use the valve for delivering metered quantities. Capsules and cartridges of, for example, gelatin for use in an inhaler or insufflator can be designed to contain a powder mix of mimetica ApoA-I and a suitable powder base, such as lactose or starch.

The composition, if desired, can be presented in a pack or in a spraying device, which may contain one or more unit dosage forms containing mimetic ApoA-I. Such packaging may include, for example, metal or plastic foil, such as in a blister pack. These packing or spray device can be provided with instructions for use.

In certain embodiments, the mimetic ApoA-I can be or be entered in the form of a complex with the lipid. Accordingly, the present invention includes mimetic ApoA-I/lipid complexes, compositions containing them and methods of their introduction. These complexes can have several advantages, as they may have an increased half-life in the circulatory line, in particular, when such a complex is the density and size similar to HDL, and particularly with populations of pre-β-1, or pre-β-2 HDL. Such is plexi convenient to receive any of a number of the following methods. Stable preparations with a relatively long shelf life, can be obtained by lyophilization, with the following way of solifenacin given here only as one of the incarnations. Lyophilized complexes can be used to obtain the mass of the material for the preparation of pharmaceutical dosage forms or to obtain individual aliquot or dosage forms which can be restored by rehydration with sterile water or an appropriate buffer solution before introduction to the subject.

To obtain these complexes can be used in a variety of ways well known to specialists in this field. For example, there may be used a number of available technologies for the production of liposomes or proteoliposomes. For example, for the formation of a complex mimetic ApoA-I may be subjected to the action of ultrasound (using baths or ultrasonic device) together with the relevant lipids. Alternatively, the mimetic ApoA-I can be combined with a pre-defined lipid vesicles with obtaining spontaneously composed of peptide-lipid complexes. In another alternative embodiment, these complexes can be formed by detergent dialysis; for example, a mixture of mimetica ApoA-I, lipid and determinatively to dialysis to remove the detergent and recovery or complex formation (see, for example, Jonas et al., 1986, Methods in Enzymol. 128: 553-582).

Alternatively, these complexes can be prepared by methods described in U.S. patent No. 6004925 ("patent '925"), a full description of which is included in the present description by reference. In the methods described in patent '925, mimetic ApoA-I and lipid are combined in a solvent system, in which jointly dissolve each ingredient and which can be removed by lyophilization. To this end choose a couple of solvents to ensure joint solubility as mimetica ApoA-I and lipid. In one of the embodiments of the mimetic ApoA-I, such a complex can be dissolved in water or an organic solvent or mixture of solvents (solvent 1). Lipid such as a phospholipid component is dissolved in water or an organic solvent or mixture of solvents (solvent 2), which is mixed with the solvent 1, and two of these solution are mixed with each other. Alternatively, the mimetic ApoA-I and lipid can be embedded in a system of mutual solvents; that is, a mixture of miscible solvents. Alternatively, the mimetic ApoA-I and lipid can be suspended in the solvent or solvent mixture. In one of the embodiments of the specified solvent mixture is a mixture of organic solvent and water. Examples of organic solvents is clucalc in itself, not limited to, acetic acid, xylan, cyclohexane and methanol. Examples of mixtures of solvents include, but are not limited to, acetic acid and xylan, acetic acid and cyclohexane, and methanol and xylan. A suitable ratio of mimetica ApoA-I and lipids may be primarily determined empirically, so that the resulting complexes had suitable physical and chemical properties; that is that usually (but not necessarily), they were similar in size with HDL. The resulting mixture is frozen and lyophilizers dry. Sometimes to facilitate lyophilization to the mixture add a further solvent. This lyophilized product can be stored for long periods, and it usually remains stable.

Alternatively, these complexes can be obtained through co-lyophilization of mimetica ApoA-I peptide in solutions or suspensions. Selected the homogeneous solution of the peptide and phospholipid in an organic solvent or mixture of organic solvents can be dried, and the peptide/phospholipid complexes can be formed spontaneously in the hydration of the lyophilized powder aqueous buffer.

Dried product can be recovered in order to obtain a solution or is Uspenie complex. This dried powder is subjected to the re-hydration of aqueous solution to the desired volume (often 5-20 mg peptide/ml, which is typical for intravenous injection). In one of the embodiments of the dried powder is subjected to the re-hydration of phosphate buffered saline, bicarbonate saline solution or saline solution. The pH of this mixture can be increased to 7.5 to 8.5. The mixture can be shaken or stirred on the vortex to facilitate rehydration, and in most cases the stage of recovery can be performed at a temperature equal to or higher than the temperature of the phase transition of the lipid component of the complex. Within minutes of receiving the drug restored the lipid-protein complexes.

You can define the properties of the aliquots of the resulting recovered drug to confirm that the complexes in the composition have the desired classification by size; for example, classification by size of HDL. Defining properties of the recovered product can be made using any method known in this field, including, but not limited to filtration by size exclusion chromatography, gel filtration chromatography, filtration column of chromatogra the iej, gel permeation chromatography and native page-electrophoresis. In one of the embodiments of the properties of recovered drug determined by gel-filtration chromatography. The size of the resulting complexes can be determined by their effectiveness. The following examples use the system gel-filtration chromatography, Pharmacia Superose 6 FPLC. The buffer that is used contains 150 mm NaCl in 50 mm phosphate buffer, pH about 7.0 to about 9, in one of the embodiments, 7.5 to 8.5, and in another embodiment to 7.4. A typical sample volume is from 20 to 200 microliters complexes containing 5 mg peptide/ml the Rate of flow of the column is 0.5 ml/min Series of proteins with known molecular weight and Stokes diameter, and human HDL are used as standards to calibrate the column of complexes of proteins and lipoproteins in relation to absorption or scattering of light with a wavelength of 254 or 280 nm.

The recovered product may also be described in relation to the definition of concentration, the final pH and the osmolality of the resulting solution, and the concentration and integrity of the peptide and individual lipids. The concentration of mimetica ApoA-I and lipid complexes can be measured by any method known in this field, including, but not limited to protein and FOS is olpidium analyses, as well as chromatographic methods such as high performance liquid chromatography ("HPLC"), gel-filtration chromatography, gas chromatography ("GC"). Chromatographs can be connected with various detectors, including, but not limited to, detectors, mass spectrometry, UV or diode array, fluorescence and elastic detectors light scattering. The integrity of mimetica ApoA-I and lipid in these complexes may be determined by the above-described chromatographic technologies, as well as by amino acid analysis, thin layer chromatography and standard tests for determining lipid oxidation.

Lipid mimetic ApoA-I/lipid complex may represent one or more different lipids including, but not limited to, saturated, unsaturated, natural and synthetic lipids and phospholipids, as well as their pharmaceutically acceptable salts. Typical salts include, but are not limited to sodium, calcium, magnesium and potassium salts.

Suitable lipids complexes mimetic ApoA-I/lipid include, but are not limited to, phospholipids (C1-C6)-alkyl chain, phosphatidylcholine (PC), egg phosphatidylcholine, soybean phosphatidylcholine, dipalmitoylphosphatidylcholine, dimyristoylphosphatidylcholine, distearoyl fatimilehin-1-myristoyl-2-palmitoyloleoylphosphatidylcholine, 1-Palmitoyl-2-myristoyltransferase, 1-Palmitoyl-2-stearoylethanolamine, 1 stearoyl-2-palmitoyloleoylphosphatidylcholine, 1-Palmitoyl-2-yearposition, 1-oleoyl-2-polymetilmetacrilate, dioleoylphosphatidylcholine, dioleoylphosphatidylcholine, dilauroylglycerophosphocholine, phosphatidylserine, phosphatidylethanolamine, phosphatidylinositol, sphingomyelin, sphingolipids, phosphatidylglycerol, diphosphatidylglycerol, dimyristoylphosphatidylcholine, dipalmitoylphosphatidylcholine, distearoylphosphatidylglycerol, dioleoylphosphatidylserine, dimyristoylphosphatidylcholine acid, dipalmitoylphosphatidyl acid, dimyristoylphosphatidylcholine, dipalmitoylphosphatidylethanolamine, dimyristoylphosphatidylcholine, dipalmitoylphosphatidylcholine, phosphatidylserine brain sphingomyelin, brain sphingomyelin, dipalmitoylphosphatidyl, distearyldimonium, phosphatidic acid, galactocerebroside, gangliosides, cerebrosides, dilauroyllecithin, (1,3)-D-mannosyl-(1,3)diglyceride, aminophenylacetic, 3-cholesteryl-6'-(glycosylation)hexylester-glycolipids and cholesterol, as well as their derivatives.

In one embodiments the lipid mimetic ApoA-I/lipid complex is a neutral phospholipid. Neutral phospholipid can be any phospholipid, which has a total charge of approximately equal to the logistics at physiological pH. In certain embodiments, the neutral phospholipid is zwitterions, which has a total charge equal to zero at physiological pH.

In another embodiment, the neutral phospholipid is a lecithin (also known as phosphatidylcholine). In certain embodiments, the neutral phospholipid is a mixture of neutral phospholipids, which contains from about 5 to about 100 wt.% lecithin. In other embodiments, the mixture of neutral phospholipids contains approximately 100 wt.% lecithin. In certain embodiments, the neutral phospholipid is a mixture of neutral phospholipids, which contains from about 5 to about 100 mol.% lecithin. In other embodiments, the mixture of neutral phospholipids contains approximately 100 mol.% lecithin.

In another embodiment, the neutral phospholipid is a sphingomyelin. In certain embodiments, the neutral phospholipid is a mixture of neutral phospholipids, which contains from about 5 to about 100 wt.% sphingomyelin. In another embodiment, the neutral phospholipid is a mixture of neutral phospholipids, which contains approximately 100 wt.% sphingomyelin. In certain embodiments, the neutral phospholipid is a mixture of neutral FOS is olpidem, which contains from about 5 to about 100 mol.% sphingomyelin. In other embodiments, the neutral phospholipid is a mixture of neutral phospholipids, which contains approximately 100 mol.% sphingomyelin.

In another embodiment the neutral phospholipid mimetic ApoA-I/lipid complex is a mixture of neutral phospholipids, which contains lecithin and sphingomyelin. The molar ratio of lecithin to sphingomyelin can vary, but typically ranges from about 20: about 1 and to about 1: about 20. In certain embodiments the molar ratio of lecithin to sphingomyelin varies from about 10: about 3 to about 10: about 6. In other embodiments the molar ratio of lecithin to sphingomyelin varies from about 1: about 20 to about 3: about 10.

In another embodiment the neutral phospholipid mimetic ApoA-I/lipid complex is a mixture of neutral phospholipids, which contains lecithin, sphingomyelin, and one or more additional neutral phospholipids. Usually more neutral phospholipid is from about 5 to about 100 wt.% of the mixture.

In another embodiment the lipid mimetic ApoA-I/lipid complex is the Wallpaper charged phospholipid. Suitable charged phospholipids include, but are not limited to, phosphatidylinositol, phosphatidylserine, phosphatidylglycerol and phosphatidic acid.

In one embodiments the lipid mimetic ApoA-I/lipid complex is a mixture of at least one neutral phospholipid and at least one of a charged phospholipid. The total number of charged phospholipid (phospholipids) in the lipid mixture can vary, but typically ranges from about 0.2 to about 10 wt.% the lipid mixture. In certain embodiments the total amount charged phospholipid (phospholipids) in the lipid mixture is about 0.2 to about 2 wt.%, approximately from 0.2 to about 3 wt.%, approximately from 0.2 to about 4 wt.%, about 0.2 to about 5 wt.%, about 0.2 to about 6 wt.%, about 0.2 to about 7 wt.%, about 0.2 to about 8 wt.% or about 0.2 to about 9 wt.% the lipid mixture. In certain embodiments the total amount charged phospholipid (phospholipids) in the lipid mixture is about 0.2, approximately 0.3, about 0.4, about 0.5, about 0.6 to, approximately 0.7, about 0.8, about 0.9 to, approximately 1.0, about 1.1, closer is Ino 1,2, about 1.3, about 1.4, about 1.5, about to 1.6, about 1.7, about 1.8 to approximately 1.9 to approximately 2,0, approximately 2.1, about 2.2, while approximately 2.3, approximately 2.4, about 2.5, about 2.6, about 2.7, about 2.8 times, around 2.9 or approximately 3.0, about 4, about 5, about 6, about 7, about 8, about 9, or about 10 wt.% the lipid mixture. The total number of neutral phospholipid (phospholipids) in the lipid mixture may also vary and may depend on the number of charged phospholipid (phospholipids) and other lipids that are included in its composition. In one embodiments the total amount of neutral phospholipid (phospholipids) in the lipid mixture is approximately approximately approximately 90 to 99.8 wt.% the lipid mixture. In one embodiments the lipid mimetic ApoA-I/lipid complex is a mixture of sphingomyelin and a charged phospholipid. In another embodiment the lipid mimetic ApoA-I/lipid complex is a mixture of sphingomyelin, dipalmitoylphosphatidylcholine ("DPPC"), and a charged phospholipid.

In one embodiments the lipid mimetic ApoA-I/lipid complex is a sphingomyelin. In another embodiment of shingami the Lin obtained from milk, eggs or brain, or obtained synthetically. In another embodiment the lipid mimetic ApoA-I/lipid complex is an analogue or derivative sphingomyelin. Suitable analogues or derivatives sphingomyelin include, but are not limited to, palmitoylethanolamide, stearoylethanolamine, D-erythrose-sphingomyelin and D-erythrose-dihydrosphingosine.

In another embodiment of sphingomyelin is artificially enriched in a particular saturated or unsaturated acyl chains. For example, milk sphingomyelin (Avanti Phospholipid, Alabaster, Ala.) has long saturated acyl chains. Sphingomyelin milk contains about 20% C16:0 (16 carbon saturated) acyl chains compared with egg sphingomyelin, which contains 80% C16:0. Using the extraction solvent may be enriched milk sphingomyelin-specific acyl chains with obtaining a composition having a concentration of acyl chains, comparable, for example, with egg sphingomyelin. Acyl chains, which can be used in the present invention include, but are not limited to, saturated acyl chains (such as dipalmitoyl, distearoyl, dialogically and cibenzoline acyl chains), unsaturated chains (such as dioscorine chain), mixed-chain saturated and unsaturated acyl CE is her (such as palmitoleate or aeoline chain), saturated and/or unsaturated chain of mixed lengths, and the ether analogues of saturated and unsaturated acyl chains.

Sphingomyelin can be synthetic, so that he had a specific acyl chain. For example, sphingomyelin milk can first of all be purified from the milk, then one particular acyl chain, for example, acyl chain C16:0, can be split and replaced by another acyl chain (such as palmitic acid or oleic acid).

Sphingomyelin can be fully synthesized, for example, through large-scale synthesis. See, for example, Dong et al., U.S. patent No. 5220043; Weis, 1999, Chem. Phys. Lipids 102(1-2): 3-12. In one embodiments the predetermined level of saturation and the composition of the fatty acids are selected from synthetic sphingomyelin.

In another embodiment the lipid mimetic ApoA-I/lipid complex is a mixture of sphingomyelin and other lipid. In this embodiment sphingomyelin, usually ranging from about 25 to about 75 wt.% this mixture.

In another embodiment, the lipid mimetic ApoA-I/lipid complex is a mixture of sphingomyelin and DPPC. In another embodiment the lipid mimetic ApoA-I/lipid complex is a mixture of sphingomyelin, DPPC and dipalmitoylphosphatidylcholine ("DPPG"). In one of the embodiments of the DPPG is present in amount of from about 0 the ome up to 10 mol.% or weight.% of the mixture. In another embodiment of the DPPG is present in amount of from about 2 to about 4 mol or weight.% of the mixture. In another embodiment of sphingomyelin and DPPG are present in the mixture in a weight or molar ratio of about 1: about 1. In another embodiment of sphingomyelin, DPPC, and DPPG are present in the mixture in a weight or molar ratio, respectively, about 1: about 1: about 0.06 to. In another embodiment of sphingomyelin, DPPC, and DPPG are present in the mixture in molar ratio, respectively, was 1.04:1:0,061. In another embodiment of sphingomyelin, DPPC, and DPPG are present in the mixture in weight ratio, respectively, 1:1:0,062. In another embodiment, the mixture contains approximately to 48.5 mol or weight.% sphingomyelin, about to 48.5 mol or weight.% DPPC and about 3 molar or weight % DPPG.

In another embodiment the complex mimetic ApoA-I/lipid complex contains one or more additional peptides. In one of the embodiments of the additional peptide is ApoA-I.

In one embodiments the weight ratio of the total number of peptide to lipid in each mimetic of Area-I/lipid complex is from about 1: about 0.5 to about 1: about 5. In another embodiment the weight ratio of the total number of peptide to lipid in each mimetic ApoA-I/lipid complex is from about 1: about the 1 to about 1: about 5. In another embodiment the weight ratio of the total number of peptide to lipid in each mimetic of Area-I/lipid complex is from about 1: about 2 to about 1: about 5. In another embodiment the weight ratio of the total number of peptide to lipid in each mimetic of Area-I/lipid complex is about 1: about to 2.5. In another embodiment the weight ratio of the total number of peptide to lipid in each mimetic of Area-I/lipid complex is from about 1: approximately 3 to 1: about 5. In another embodiment the molar ratio of the total number of peptide to lipid in each mimetic of Area-I/lipid complex is from about 1: about 2.5 to about 1: about 20. In another embodiment the molar ratio of the total number of peptide to lipid in each mimetic of Area-I/lipid complex is about 1: about 9,2.

When the lipid mimetic ApoA-I/lipid complex is a mixture of sphingomyelin, DPPC and DPPG, the weight ratio of peptide: sphingomyelin: DPPC: DPPG usually is, respectively, about 1: about 1: about 1: about 0,08. In one embodiments the weight ratio of peptide:sphingomyelin:DPPC:DPPG is, respectively, 1:1,2125:1,2125:0,075. The molar ratio of peptide: sphingomyelin:DPPC:DPPG is, accordingly, approximately 1 to approximately 4: approximately 4: about 0.03. In one embodiments the molar ratio of peptide: sphingomyelin:DPPC:DPPG is, respectively,, 1:4,55:4,36:0,27.

In another embodiment, the mimetic ApoA-I/lipid complex contains from about 40 to about 85 wt.% lipid and from about 15 to about 60 wt.% the peptide.

In another embodiment, each mimetic ApoA-I/lipid complex is from about 2 to about 12 nm in diameter.

V. Methods of treating or preventing the condition

Although the inventors do not wish to bind themselves to any particular theory, nevertheless they believe that the spiral formed by the mimic ApoA-I according to the invention, exactly mimics the structural and functional properties of amphipatic helical regions of native ApoA-I, which are important for the implementation of the activation binding of lipid outflow of cholesterol and/or activation of LCAT, thus leading to the formation of peptides that exhibit high ApoA-I-like activity. In one of the embodiments of the mimic ApoA-I function through the formation of amphipatic helices (in the presence of lipids, lipid binding, formation of pre-β-like or HDL-like complexes, activation of LCAT, increase the concentration of HDL in serum and activation outflow of cholesterol.

In the bottom of the embodiments, the mimetics ApoA-I activate LCAT. In another embodiment, the mimetics ApoA-I does not activate LCAT. In another embodiment, the mimetics ApoA-I activates LCAT, but only to such extent, which does not accelerate the esterification of cholesterol. In another embodiment, the mimetics ApoA-I activates LCAT and, thus, accelerate the esterification of cholesterol, but at the same time accelerating the esterification of cholesterol only due to the activation of LCAT is insufficient for treating or preventing the condition.

In one of the embodiments of the present invention is related to methods of treating or preventing a condition involving the administration to a mammal, in case of need, an effective amount of mimetica ApoA-I.

Examples of dyslipidemia include any disorder in which increasing the concentration of HDL in serum LCAT activation and activation of the outflow of cholesterol and RCT are beneficial. Such disorders include, but are not limited to, hyperproteinemia (such as hyperchylomicronemia), a high concentration of LDL in the serum, a high concentration of VLDL in serum, hyperlipidemia (such as hypercholesterolemia or hyperglyceridemia (such as hypertriglyceridemia)), low concentration of HDL in serum, hypocholesterolemia, A-beta lipoproteinemia, deficiency of ApoA-I and Tangier disease.

Examples of cardiovascular diseases include SEB is, but not limited to, metabolic syndrome, coronary heart disease, atherosclerosis, restenosis (e.g., prevention or treatment of atherosclerotic plaques that develop as a result of medical procedures, such as balloon angioplasty), groove toxins (which is often the result of septic shock), congestive heart failure (such as chronic or acute heart failure), circulatory shock, cardiomyopathy, heart transplantation, myocardial infarction, cardiac arrhythmia (such as atrial fibrillation), supraventricular tachycardia, atrial flutter, paroxysmal atrial tachycardia, aneurysm, angina, acute cerebrovascular disease (stroke), peripheral vascular disease, cerebrovascular disease, renal disease, atherogenesis, atherosclerosis, acute pancreatitis, and coronary arteries.

Endothelial dysfunction is any imbalance between vasodilating and vasoconstricting factors and any abscopal growth and stimulating the growth factors produced by the endothelium. Endothelial dysfunction is usually weakens the ability of blood vessels to relax.

Examples of macrovascular disorders include any disorder major blood vessel. Such resstr istwa include, but not limited to, transient ischemic stroke, stroke, angina, myocardial infarction, heart failure and peripheral vascular disease.

Examples of microvascular disorders include any disorder of the small blood vessel. Such disorders include, but are not limited to, diabetic retinopathy (non-proliferative, proliferative, macular Eden), microalbuminuria, macroalbuminuria, end-stage renal disease, erectile dysfunction, autonomic neuropathy, peripheral neuropathy, osteomyelitis and ischemia of the lower limb.

The mimic ApoA-I can be entered separately or in combination with one or more other means, which are applicable to the treatment condition. Such treatment includes, but is not limited to, simultaneous or sequential introduction of the considered pharmaceuticals.

In one of the embodiments of the methods of treating or preventing the condition can additionally provide for the introduction of one or more drugs from one or more of the following classes: ACE inhibitors (angiotensinase enzyme), beta-blockers, nitrates, calcium channel blockers, diuretics, thrombolytic agents and tools that reduce ur the level of cholesterol in the blood. In another embodiment, the methods of treating or preventing the condition additionally provide for the introduction of one or more of the following medications: cholestyramine, colestipol, colesevelam, gemfibrozil, ciprofibrate, clofibrate, fenofibrate, bezafibrat, ezetimibe, ramipril, verapamil, nicardipine, diltiazem, carvedilola, nadapal, isosorbide of Mononitrate, propranolol, isosorbide dinitrate, digoxin, furosemide, metoprolol tartrate, trandolapril, nitroglycerin, amlodipine besilate, oxycodone, clopidogrel, nifedipine, atenolol, lisinopril, empirin and lanoxin.

In another of the embodiments, methods of treating or preventing the condition can further include introducing one or more lower the cholesterol drugs known to experts in the field; for example, biliary acids, Niacin, and/or statins, such as atorvastatin, simvastatin, pravastatin, fluvastatin and pitavastatin. This treatment may be particularly beneficial therapeutic effects, because each drug acts on a different target in the synthesis and transport of cholesterol; that is, the biliary acids act on recycling of cholesterol on the population of chylomicrons and LDL; Niacin before vsegdastoboy on the population of VLDL and LDL; statins inhibit the synthesis of cholesterol, reducing the population level of LDL (and possibly increasing the expression of LDL receptor); whereas the mimic ApoA-I act on RCT, increase HDL levels, increase the activity of LCAT activity and outflow of cholesterol.

In another embodiment, the methods of treating or preventing the condition can additionally include the introduction of fibrate, such as clinofibrate, semifinal, fenofibrate and benzafibrate.

In another of the embodiments, methods of treating or preventing the condition can include the introduction of antimicrobial agents and/or anti-inflammatory agents, for example, which is used for the treatment of septic shock caused by endotoxin.

The mimic ApoA-I, you can enter any of the suitable ways that provide bioavailability in the circulation. This can be achieved by parenteral routes of administration, including intravenous (IV), intramuscular (I/m), intradermal, subcutaneous (s/C) and intraperitoneal (/b) injection. However, there may be used other routes of administration. For example, absorption through the gastrointestinal tract can be achieved by the oral route of administration (including, without limitation, swallowing, buccal and sublingual route), provided that use of those compositions (e.g. the R, with InterCasino coatings)to avoid or minimize the degradation of peptides, for example, in the harsh conditions of the mucous membrane of the mouth, stomach and/or small intestine. Alternatively, it may be used for introduction through tissue of the mucous membrane, such as vaginal and rectal routes of administration, in order to avoid or minimize degradation in the gastrointestinal tract. As another alternative, the composition according to the invention can be introduced percutaneous (e.g., transdermal), eye, by, or by inhalation. Professionals should be clear that the chosen route of administration may vary, depending on the condition, age and compliance of the recipient.

The actual applied dose mimetica ApoA-I may vary depending on route of administration and can be adjusted with the aim of achieving concentrations mimetica ApoA-I in plasma, comprising from 100 mg/l to 2 g/l In one of the embodiments of the dose mimetica ApoA-I correct in order to achieve the level of free mimetica ApoA-I or part of a complex mimetica ApoA-I in the serum for at least 24 hours after administration of about 10 mg/DL 300 mg/DL higher than the base (source) the level prior to the injection.

The mimic ApoA-I can be administered in a variety of therapeutic schemes. In one of the embodiments of the mimetic ApoA-I is injected PU is eat injection in a dose of 0.5 mg/kg to 100 mg/kg once a week. In another embodiment the required levels in serum may be maintained by continuous infusion or by intermittent infusion, providing from about 0.5 mg/kg/HR to 100 mg/kg/hour mimetica ApoA-I. In one of the embodiments of the mimetic ApoA-I is administered at a dose of approximately 20 mg/kg

In another embodiment of the mimetic ApoA-I is administered by intravenous injection one or more times a day. In another embodiment of the mimetic ApoA-I is administered by injection once every 3 to 15 days, once every 5-10 days or once every 10 days. In another embodiment of the mimetic ApoA-I is administered as a series of supporting the injection, the series supports injection administered once every 6 months to one year. Series supports injection can be entered, for example, during the whole day (perfusion to definitely maintain a set level of complexes in the plasma), several days (for example, four injections during the eight-day period) or several weeks (for example, four injections over a four-week period).

In another of the embodiments can be entered increasing doses of mimetica ApoA-I, ranging from approximately 1 to 5 doses in the amount of approximately 50 mg to approximately 200 mg / introduction, with subsequent repeated doses of from about 200 mg to about 1 g of the introduction. Depending on the needs of the patient, the introduction may be effected by slow infusion lasting more than one hour, by rapid infusion lasting one hour or less, or by a single bolus injection.

Toxicity and therapeutic efficacy of mimetics of ApoA-I can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, with the definition of the indicators LD50(the dose lethal to 50% of the population) and ED50(the dose therapeutically effective in 50% of the population). The ratio of the dose, which has toxic and therapeutic effects is a therapeutic index, and it can be expressed as the ratio LD50/ED50. In one of the embodiments of the mimic ApoA-I have a wide therapeutic index.

VI. Methods of analysis

The mimic ApoA-I can be investigated in relation to their ability to form α-helix in the presence of lipids, to contact with lipids to form complexes with lipids, activate LCAT, activate the outflow of cholesterol and other

Methods of analysis for the study of the structure and/or function mimetics of ApoA-I is well known in this field. Several of these ways are listed below. For example, in the following examples, the methods of the circular, or circular dichroism (CD) and I the cluster magnetic resonance (NMR) can be used to analyze the structure mimetics of ApoA-I and in particular, the degree of helicity in the presence of lipids. The ability to bind to lipids can be determined using fluorescence spectroscopic analysis described below in the examples. The ability of the peptides and/or peptide analogs activate LCAT can be easily determined using the LCAT activation method described below in the examples. Testsin vitroandin vivodescribed below in the examples, can be used to estimate the time half-life, distribution, outflow cholesterol and effects in relation to RCT.

In General, the mimic ApoA-I according to the invention, which exhibit the properties listed below in table 10, are particularly effective.

Table 10
The range 1Range 2
% helicity in the presence of lipids (Ri=30) (non-blocked peptides, with a 22 amino acid residue)≥60%≥80%
% helicity in the presence of lipids (Ri=30) (non-blocked peptides with 18 amino acid residues)≥40%≥60%
% helicity in the presence of lipids (Ri=30) (blocked peptides having 18 or less amino acid residues)≥60%≥80%
The binding of lipids (in the presence of small unilamellar vesicles (SUV)The peptide in 0.5-10 μm (Ri=1-50)
Activation of LCAT≥38%≥80%
Riis a lipid:peptide molar ratio.

The ability mimetica ApoA-I to form an α-helix in the presence of lipids can be demonstrated using the following method CD. Those peptides that spirality by at least 40% (non-blocked peptides having 18 or less amino acid residues) or spirality at least 60% (blocked peptides having 18 or less amino acid residues; non-blocked peptides having 22 or more amino acid residue) and which are associated with lipids (concentration, component approximately 5 μm and the lipid:peptide molar ratio, constituting approximately 30), in particular, those mimic ApoA-I, which contain a fluorescent residue Trp (W) or Nal, can be identified is the following fluorescence analysis. However, for mimetics of ApoA-I that do not contain fluorescent residues, binding to lipids is observed when the degree of helicity in the presence of lipids is increased.

In one of the embodiments of the present invention mimic ApoA-I, in particular, those which exhibit the property of binding to lipids in the presence of SUV (peptides in 0.5-10 μm; with the lipid:peptide molar ratio in the range from 1 to 50), is subjected to screening to determine their ability to activate LCAT, since the peptides are able to activate LCAT, are particularly effective in the ways described here. In one of the embodiments of the mimetics of ApoA-I appears at least approximately 38% ability to activate LCAT compared with native human ApoA-I (which is determined using the analysis described here activate LCAT). In another embodiment of the mimetics of ApoA-I appears at least approximately 50%, 60%, 70%, 80% or even 90% ability to activate LCAT compared with native human ApoA-I.

VII. Other applications

The mimic ApoA-I are used in the analysis ofin vitrofor determination of serum HDL, for example, for diagnostic purposes. Since it is generally mimic ApoA-I associated with HDL-component of serum, such mimetics can be used as "markers" for the population of HDL. Accordingly, the present the invention relates also to methods of measurement of serum concentrations of HDL, providing reduction of serum HDL in contact with a number of mimetica ApoA-I, which is associated with serum HDL, and calculate the amount of ApoA-I-associated HDL. Moreover, the mimic ApoA-I can be used as "markers" for subpopulations of HDL, which is effective in the reverse transport of cholesterol ("RCT"). To this end, the mimetic ApoA-I can be added or mixed with the serum sample of a mammal containing HDL; after an appropriate time of incubation component of HDL can be investigated by detecting the embedded mimetica ApoA-I. This can be done using labeled mimetica ApoA-I (for example, radioactive labels, fluorescent labels, enzyme labels, dyes, etc. or using immunoassays using antibodies (or fragments of antibodies)that are specific in relation to mimetica ApoA-I.

Alternatively, the labeled mimic ApoA-I are used in imaging procedures (e.g., CAT-scans, MRI scans) to visualize the circulation system or to monitor an RCT, or to visualize the accumulation of HDL in fat strips in areas of atherosclerotic plaques, etc. (where the paps must be active in the outflow of cholesterol).

Further, the present invention includes the following non-limiting illustrative examples.

Examples

Example 1

Synthesis of mimetics of ApoA-I

The mimic ApoA-I is produced by solid-phase peptide synthesis (PPPS) using Fmoc chemistry(9-fertilityscore). C-terminal residue covalently associated with 4-methylbenzhydrylamine (MBHA) resin. Then attach the remaining amino acid residues by cyclically repeating the removal of the protective Fmoc group and attaching a protected amino group. After solid-phase Assembly of the peptide, this peptide is separated from the resin triperoxonane acid (TFU). The crude peptide was obtained by precipitation and drying. The identity of the crude peptide was confirmed by mass by a spectroscope and by amino acid analysis.

Example 2

Cleaning mimetics of ApoA-I

Cleaning mimetics of ApoA-I was made in accordance with example 1 by preparative obremeniaet HPLC stationary phase C18 (grafted silicon dioxide, the particle size of 15 μm, pore size 120 Å) using a gradient of water/acetonitrile (0.1% counterion TFU). Eluruumina fractions were detected by UV absorption at 220 nm. The result of each run is approximately 15 g of the crude peptide purified fractions are collected and concentrated on a rotary evaporator. Then a solution of the peptide is purified on a column of C18 HPLC used in the first stage of purification. After this race the thief peptide concentrate on rotoroa evaporator to remove acetonitrile and lyophilizers.

Next, the dried peptide powder again solubilizer in a mixture of 90% water/10% acetonitrile, and in the process of ion-exchange chromatography the counterion was exchanged for acetate (resin Dowex, eluting environment 90% water/10% acetonitrile). Purified peptide from the acetate counterion filtered through a sterile membrane with a pore diameter of 0.22 micrometer and lyophilizers.

Example 3

Synthesis of peptide 16

Peptide 16 was synthesized on solid-phase substrate using Fmoc chemistry(9-fertilityscore). C-terminal isonicotinamide the residue was covalently linked to the resin linker type of linker Wang (4-methylbenzhydrylamine (MBHA) resin). Used protective groups of amino acids were as follows: tert-bucilina group in the case of Glu and Asp, Boc-group in the case of Lys, Pbf-group in the case of Arg, Trt-group in the case of Asn and Gln.

Solid-phase Assembly of peptides was carried out manually in the reactor 601, equipped with a porous disk with a mechanical stirrer and a device for ozonation nitrogen. Resin, para-methylbenzhydrylamine resin (polystyrene-l%-divinylbenzene), soaked and washed with dichloromethane (DCM)/dimethylformamide (DMF) (95/5). Embedding the C-terminal residue was achieved by linking the C-terminal isonipecotic acid esterified on the linker MPPA (linker type of linker Wang). The binding reaction was performed is of 1.35 EQ. Fmoc-Inp-MPPA-linker, 1,35 EQ. N-hydroxybenzotriazole (HOBt), benzotriazol-1-yl-oxtriphylline of hexaflurophosphate (PyBOP) and 4,05 EQ. diisopropylethylamine (DIEA) in DMF/DHM(50/50). After binding, the resin 3 times washed with DMF.

The peptide chain was assembled on the resin by cyclically repeating the removal of the protective Fmoc group and attaching a protected amino group. All amino acid residues were added, following the same cycle: first, the protective Fmoc group was removed in piperidine (35% in DMF) in three repeated cycles. (Reaction removal of the Fmoc protection was approximately 16 min). After removal of the protective Fmoc group, the resin in nine repeated cycles washed with DMF. Then Fmoc-protected amino acid residues (2 EQ.) connected with 2 EQ. N,N'- diisopropylcarbodiimide (DIC)/HOBt in a mixture of DMF and DHM (50/50). (Stage binding took approximately one hour to a full day). In order to determine over whether the reaction is the binding used ninhydrin test. If the ninhydrin test indicates that the binding reaction is not completed, the stage of binding was repeated with a smaller excess (0.5 to 1 EQ.) amino acid, PYBOP, HOBt in DMF/DHM and DIEA. After the stage of binding resin washed with DMF three repeated cycles.

Then the peptide was separated from the resin and remove the protective group. Separation from the resin and removal of the protection was carried out by the parties in a mixture of T Is U/water/anisole (90/5/5.about./about.) at a concentration of 5 ml/g of the peptide to the resin for 2.5 hours at room temperature. In the process of gradually adding resin to the mixture of the reactants the temperature was regulated so that it remained below 25°C. the Peptide was soluble in TFU and was extracted from the resin by filtration through a porous disk. After concentration on a rotary evaporator, the peptide was subjected to precipitation in cold methyl tert-butyl ether (MTBE) (0±5°C) and filtered. The crude peptide is washed with MTBE and dried under reduced pressure in an oven at 30°C.

After removing the last of the protective group Fmoc peptide was treated with TFU/H2O for cleavage and removal of the protective groups of the side chains. Then the crude peptide was besieged from cold ether and collected by filtration.

Example 4

Purification of the peptide 16

Peptide 16, obtained in accordance with example 3, was purified by preparative obremeniaet HPLC (high performance liquid chromatography) with a stationary phase C18 using a gradient of water/acetonitrile (counterion TFU). Prochrom column LC110 stuffed new or specialized stationary phase C18 (grafted silicon dioxide, the particle size of 15 μm, pore size 120 angstroms). The packing of the column was controlled by SST for a certain number of layers, and asymmetry factor of the peak.

On a Prochrom column LC110 number of peptide injectable for each run, were approximately $ 15 Gneisenau peptide, dissolved in water/acetonitrile (80/20) at a concentration of approximately component of 75 g/liter was passed Through the column, the gradient of buffer B in buffer A (flow rate approximately 450 ml/min and UV-detection at 220 nm): buffer A=0.1% OF TFU in water; buffer B = acetonitrile/0.1% of TFU in water (80/20), under the following conditions:

Column:Symmetry C18, 5 μm, 250 x 4.6 mm, 100 Å
Gradient:40% buffer B to 55% buffer B for 30 minutes
when the speed of the current of 1 ml/min
Temperature:60°C
Detection:210 nm

Eluruume fractions were analyzed by analytical HPLC and collected in four categories: depleted, the front impurity, pure and rear impurity, in accordance with pre-established standards. Established standards for purity in progress HPLC to classify fractions by category are as follows:

Depleted:<80%
Net:≥95%
Lane is dnaa and rear impurity: From ≥80 to < 95%

To ensure greater product yield impurity fractions adjacent to the net (the front of the impurity and the rear impurity) was subjected to repeated passes through the same column. Clean pools concentrated on a rotary evaporator to remove acetonitrile.

Example 5

The exchange of counterions and drying of peptide 16

Clean pools from example 4 was mixed and placed on a stirrer for homogenizing. Concentration of the pure peptide 16 was taken using obremeniaet HPLC on a preparative column, which was used for purification. On a Prochrom column LC110 net peptide injected for each run, were approximately $ 20 l at a concentration of approximately component 5 g/liter was passed Through the column, the gradient of buffer B in buffer A (flow rate approximately 450 ml/min and UV-detection at 220 nm): buffer A=0.1% OF TFU in water; buffer B = acetonitrile/0.1% of TFU in water (80/20). Through the column missed a step gradient of buffer B in buffer A (flow rate approximately 450 ml/min and UV-detection at 220 nm): buffer A=0.1% OF TFU in water; buffer B = acetonitrile/0.1% of TFU in water (80/20).

The volume of solvent for flushing full peak was collected, concentrated on a rotary evaporator to remove acetonitrile and liofilizirovanny on the bottle whether the filestore. The resulting freeze-dried pools of purified peptide was mixed in water/acetonitrile (90/10) at a concentration of 80 g/l and stirred until complete dissolution before ion-exchange chromatography on acetate Dowex strictly mainly with resin 50-100 mesh. (Acetate Dowex was obtained by processing the resin Dowex C1 1N NaOH, then rinse with purified water processing AcOH/H2O (25/75) and washing with purified water.) The sample was suirable water/ acetonitrile (90/10). The volume of solvent for flushing full peak was collected and concentrated on a rotary evaporator if lucindy volume was too large. The purified peptide solution was filtered through a sterile filter capsule (0.22 micrometer) and liofilizirovanny on shelf-liofilizadora.

Example 6

Analysis of the purity of the peptide 16

The purity of the peptide 16 was determined using analysis by analytical HPLC with reversed phase. The purity was determined by integration of the areas of all peaks (normalized square). The analysis was undertaken using system Waters Alliance HPLC with module 2695, consisting of a pump with double valve reciprocating movement, a degasser, an automatic injection system with adjustable column thermostat Peltier; UV-detector module 2487; and software Empower Pro Version 5.00. The used column was a column Symetry C18 (5 μm) or equivalent column, 250×4,6 mm column Temperature was 60°C. Injections were suirable in a gradient profile with the velocity of the current, part 1 ml/min Eluent A was 0.1% of TFU (for example, Acros 13972) in water milli-Q, while the eluent B is 0.1% of TFU in acetonitrile in the gradient category HPLC (e.g., SDS 00637G). The gradient profile is shown below:

Time (min)Eluent A (%)Eluent B (%)
0,05743
30,05050
45,02080
46,00100
51,00100
52,05743

Peptide 16 was detected by UV absorption at 210 nm. The time course was 45 min with a retention time of 22 min between injections and leaching from the column. Peptide 16 outweighed in the HPLC vial and was dissolved in purified water to obtain concentrate the radio approximate component 1.2 mg/ml were injected with 20 µl of the peptide solutions.

Example 7

Determination of properties of mimetics of ApoA-I by LC-MS (liquid chromatograph-mass spectrometer)

Standard commercially available three-stage quadrupole mass spectrometer (model TSQ 700; Finnigan MAT, San Jose, Calif., USA) was used to determine the mass. Interface with pneumatically equipped with elektrorazpredelenie (ESI) was used for introduction of the sample into the ionization source at atmospheric pressure mass spectrometer. Interface [interface] the sprayer worked at a positive potential of 4.5 kV. The steel temperature of the capillary was maintained at 200°C, whereas the temperature of the distributor was 70°C. Positive ions produced in the result of a process occurring in this ion evaporator flow to the analyzer of the mass spectrometer. The multiplier is set at 1000 C. the Analyzer compartment mass spectrometer installed on 4E-6. All data were obtained at a resolution of <1 micron.

The mimic ApoA-I were analyzed by direct infusion of purified mimetics of ApoA-I using system microcolony ABI (Applied Biosystems)consisting of a syringe pump (model 140B), UV detector (model 785A) and oven/injector (model 112A). The solvent system consists of water (solvent A) and acetonitrile (dissolve the spruce B), each of which contains 0.1% of the TFU. The mimic ApoA-I infuziruut either gradient or isocratic conditions and elute from the column Aquapore C18. The current velocity is usually 300 ál/min, the Concentration of each mimetica ApoA-I is approximately 0.03 mg/ml, 20 μl of which is injected (for example, 30 pmol).

MS-experiments full scan obtained by scanning quadrupole 1 from m/z 500-1500 4's. Data were obtained using the DEC Alpha system and were processed using software provided Finnigan MAT (BIOWORKS).

Example 8

Defining properties of peptide 16 by LC-MS

Mass spectral analysis was performed using instrumentation Thermo-Finnigan LCQ Advantage. Source served as electrospray ionization (ESI-MS). The MS parameters: gas flow of nitrogen = 30 conventional units, the voltage in the electrostatic spraying = 5.2 V, the temperature in the capillary = 270°C, the tension in the capillary = 38, on the lens in the knee cathode-ray tube (Tube Lens Offset)=55 C. the Test solution of 100 μg/ml solution of peptide 16 was analyzed in a solution of methanol/water/formic acid 47/47/6 about./about./about. (direct infusion into the MS at a speed of injection of 5 μl/min through a 500 ál syringe). The result obtained after inverse filtering, coincided with theoretical value.

Example 9

Linakis is now analysis mimetics of ApoA-I

Amino acid analysis was made using the amino acid analyzer ABI 420 (Applied Biosystems). This system consists of three modules: gidrolizuemye and derivatizing tool, obremenitve HPLC and data systems. Samples of the peptides was applied 3 times in three repetitions) on porous glass slides and consistently subjected to hydrolysis in the gas phase (155°C, 90 min). After removal of HCl, the resulting amino acids were converted into PTC-AA (phenylthiocarbamoyl-amino acids) using PITC (phenylisothiocyanate). After transfer of the sample into the cell WAH the resulting mixture was fractionally in column (Aquapore C18 in gradient mode (solvent A: 50 mmol ammonium acetate (NH4Ac), pH of 5.4, in water; solvent B: 32 mmol of sodium acetate (NaOAc) in aqueous acetonitrile) under conditions of controlled temperature. Data HPLC were processed using the software package provided by Applied Biosystems. Quantitative evaluation was performed by relatively standard peptides supplied by Applied Biosystems.

Example 10

Amino acid analysis of peptide 16

Peptide 16 (about 700 g) hydrolyzed in 100 Microlitre 6N HCl (e.g., Pierce 24308) at 110°C for 20 hours on a composite amino acids after derivatization were separated and quantitatively evaluated against STD is bound mixture of amino acids (amino acid standard H, for example, Pierce 20088). Amino acids were derivateservlet using o-phthalaldehyde (OPA-reagent, for example, Fluka 5061-3335) and 9-fertilityrate (Fmoc-reagent, for example, Fluka 5061-3337), and then were injected with column C-18 HPLC. Used for the analysis Agilent 1100 HPLC with UV detector and Chemstation software. The used column was a column Hypersil ODS 200×2.1 mm, 5 μm. Used the gradient consisted of 0-60% B in 17 min to 100% B for 7 min at the speed of the current of 0.45 ml/min (Buffer A=2.3 g of sodium acetate in 1000 ml of H2O + 180 μl of triethylamine, pH brought to 7.2 with 2% solution of acetic acid + 3,3 ml of tetrahydrofuran. Buffer=2.3 g of sodium acetate in 200 ml of H2O, the pH was brought to 7.2 with 2% solution of acetic acid and 400 ml of acetonitrile and 400 ml of methanol. Evaluation of the amino acids was carried out in three repetitions, and the amino acids were detected by UV absorption at 368 and 262 nm. Standard solution Pierce were injected with both before and after the injection of the peptide sample in three parallel samples.

Example 11

Obtaining the peptide/lipid complexes by co-lyophilization

50 mg mimetica ApoA-I was dissolved in 1 ml of glacial acetic acid in 1 ml clear glass vial with a cap. Dissolving the peptide facilitated periodic stirring on a vortex for 10-minute periods at room temperature. 50 m is dipalmitoylphosphatidylcholine (DPPC; lipids Avanti Polar, 99% purity, product #850355) and 50 mg of egg sphingomyelin (NOF) was dissolved in 1 ml of glacial acetic acid. DPPG was dissolved in a mixture of 90% glacial acetic acid and 10% water (vol./about.) at a concentration of 10 mg/ml Dissolution DPPG was facilitated by incubation at 37°C. Mimetic ApoA-I, sphingomyelin, solutions of DPPC and DPPG were mixed with obtaining weight ratio mimetic ApoA-I:sphingomyelin:DPPC:DPPG, constituting, respectively, of 1:1.35:1,35:0,30. The resulting solution was frozen at -20°C and liofilizirovanny for more than 12 hours.

Dried powder was subjected to hydration in bicarbonate buffered saline (20 mm sodium bicarbonate, 130 mm NaCl, pH 8,2) to obtain the final concentration of mimetica ApoA-I containing 10 mg/ml and the Mixture was stirred to facilitate rehydration. After hydration, the pH was brought to a pH of 7.4, using a solution of 1H. NaOH. To facilitate the complexation hydrated powder was incubated in a water bath at 50°C for 15 minutes, followed by keeping at room temperature for 15 minutes. Heating and cooling was repeated until then, until he got obtaining clear transparent solution mimetic ApoA-I/phospholipid complexes in the buffer.

Example 12

Obtaining the peptide 16/lipid complex by homogenizing

Received sodium phosphate buffer (12 mm, pH 8,2) and heated it in the up to 50°C.

The variance of DPPG was obtained by dispersing DPPG in the buffer at a concentration of 45 mg/ml Solution of peptide was obtained by dissolving the peptide 16 in the buffer at a concentration of 30 mg/ml. pH of the peptide solution was brought about to 8.2 by adding NaOH. Then the peptide solution was combined with the dispersion of DPPG and incubated at 50°C until then, until he got to obtain a clear solution of the peptide/DPPG.

The variance of the sphingomyelin/DPPC was obtained by dispersing sphingomyelin and DPPC in the buffer at a concentration of 38.3 mg/ml of each component, sphingomyelin and DPPC. Then the variance of the sphingomyelin/DPPC mixed by intense horizontal shaking, stirring.

Solutions of peptide/DPPG and sphingomyelin/DPPC were combined and homogenized using a high-pressure homogenizer (Avestin C3), up until the solution became transparent and did not begin to form complexes. After homogenization was added isotonic agent (130 mm NaCl). Then the solution was sterilized by passing through a sterile filter and filled them with glass bottles. The final concentration of peptide 16 in solution was 15 mg/ml

Example 13

Analysis of the peptide 16/lipid complex

a. The distribution of complexes by size

Confirmed the identity of the peptide 16/lipid complex obtained in accordance with the leader 12, and distribution complexes by size were determined using gel permeation chromatography (GPC). For the separation used a Tosoh TSK-GEL G3000SWXL(ID 7.8 mm, length 30 cm). Injections were suirable 6 mm phosphate buffer containing 150 mm NaCl (pH 7,4), and isocratic speed of the DC component of 1 ml/min, the Samples were obtained by 20x dilution with mobile phase, and use the volume of injection was 100 µl. Throughput columns tested before each application by injection four standards with known molecular weights. The complex was detected by UV at a wavelength of 220 nm. The identity was confirmed by comparing the retention time of the complex with the reference standard. The distribution of complexes by size expressed as a percentage of total peak in the chromatogram. GPC-chromatogram for the peptide 16/lipid complex obtained in accordance with example 12, are presented in figure 5.

b. Identity, purity and content of peptide 16 complex

Identity, purity and content of peptide 16 complex were determined using ultra-efficient liquid chromatography ("UPLC") with UV detection at a wavelength of 215 nm. For this separation used a 100 mm column Acquity BEH C18 with I.D. 2.1 mm, particle size 1.7 μm. Injections were suirable using a mobile phase BINAR is on a gradient of 0.1% (vol./about.) TFU when the ratio of methanol/acetonitrile/water as 52,5/22,5/22/about./about. and 0.1% (vol./about.) TFU when the ratio of methanol/acetonitrile/water as 56/24/20 about./about./about. The samples were obtained by 20x dilution and were injected with using the amount of injection of 7.5 µl. The combination of the organic solvent mobile phase contributed to the dissolution of the complexes and separation of peptide 16 from lipid complex. The identity of the peptide 16 was confirmed by comparing its retention time with a reference standard. The purity of the peptide 16 was expressed as a percentage of total peak in the chromatogram. The content of peptide 16 was calculated using a calibration curve based on reference standards diluted solutions of peptide 16.

c. Determination of lipid content in the complex

The lipid content of the peptide 16/lipid complex obtained in accordance with example 12, were determined using enzymatic analysis using the method of DAOS. The test kit is manufactured by Wako Pure Chemical Industries, Ltd. (Phospholipids C kit). The samples were dissolved in 75x, using phosphate buffer. Enzymes in the specified collection for analysis of sphingomyelin hydrolyzed and DPPC with the release of choline, which then reacts with several other enzymes, activating the blue pigment. The blue pigment was determined spectrophotometrically. Conducted quantitative determination of samples from the calibration curve obtained on on the basis of dilutions cholate sodium and blue pigment. Hydrolyzed sphingomyelin and DPPC contained choline and, thus, their expected by the specified method.

Example 14

Gel-filtration chromatography of human HDL on superose 6

Human HDL2(HDL2) is obtained as follows: 300 ml frozen human plasma (Mannheim Blutspendzentrale #1185190) were subjected to thawing, its density brought up to 1.25 with solid potassium bromide, and centrifuged for 45 hours at 40,000 rpm in a Ti45 rotor (Beckman) at 20°C. the resulting unbound upper layer was collected, were dialyzed against distilled water, the density brought to 1.07 with solid potassium bromide, and centrifuged as described above for 70 hours. The bottom layer (at the level of 1 cm from the bottom of the tube) were collected, added to 0.01% sodium azide, and this layer was kept at 4°C for 4 days. 20 μl of HDL2put on superose 6, Pharmacia, gel filtration chromatography, using 0.9% NaCl as a column of the eluate. The flow velocity through the column was 0.5 ml/min monitored column of the eluate by the absorption or scattering of light at a wavelength of 254 nm. A series of proteins with known molecular weights and diameters Stokes equations were used as standards to calibrate the column to calculate the Stokes diameters of the particles (Pharmacia Gel Filtration Calibratin Kit Instruction Manual, Pharmacia Laboratory Separation, Piscataway, N.J., revised April 1985).

Example 15

Determination of peptide 16 in plasma in rats and monkeys using precipitation by detection using liquid chromatography and tandem mass spectrometry (LC-MS/MS)

The concentration of peptide 16 was determined in the plasma of rats or monkeys in the concentration range from 1 to 500 mcg/ml using a blank sample matrix. Labeled isotope peptide 16 was used as a solution for the internal standard and was added to the thawed plasma samples. Then these samples were besieging protein using water: acetonitrile: TFU (70:20:10./about./vol.), followed by mixing and centrifugation. The supernatant was transferred into a clean 96-well plate, and each well was added water: acetonitrile: TFU (70:30:0.1 to about./about./vol.), followed by mixing and centrifugation before analysis LC-MS/MS. Conditions LC (LC) were as follows: Acquity UPLC and a turbo ion-spray mass spektroskopiya (positive ion (MS/MS), using column BEH Shield RP18 noise gradient of water:acetonitrile: TFU 0,1%.

The concentration of peptide 16 in the calibration standards and QC samples were determined using linear regression based on the principle of least squares, inverse concentration (1/x)as the weighting factor.

Example 16

Measurement is their pharmacokinetics of the peptide 16/lipid complex in rats

The pharmacokinetics of the peptide 16/lipid complex (where the lipids are represented by sphingomyelin, DPPC, and DPPG in a weight ratio of 1:1,2125:1,2125:of 0.075, and the peptide: lipid weight ratio is 1:2,5) were evaluated in rats Windstar.

Nine animals of each sex per group were taken for evaluation of pharmacokinetics. Animals in the control group media received intravenous 20 ml/kg 130-millimolar sodium chloride, 12 mm phosphate buffer, pH of 8.2. Animals in groups treated peptide 16/lipid complex, introduced it by 15, 30 or 60 mg/kg every other day by intravenous infusion. Approximately 0.5 ml of blood was collected from retroorbital sinus under isoflurane anesthesia and collected in tubes containing Na3EDTA as anticoagulant, cohort 3 animals in the cohort at the starting point and through 0,0833, 0,5, 1, 2, 6, 12, 24 and 48 hours after the dose on day 0 and day 26. Thus, each cohort of animals, the blood was collected at three different time points. Plasma was separated by subsequent centrifugation and frozen at -20°C until the analysis. The levels of the peptide was analyzed by the method LC-MS/MS as described in example 8. Pharmacokinetic parameters of individual concentrations in plasma were determined by pharmacokinetic analysis without the compartmentalization using kinetics 4.4.1. After centuries the Denia dose levels of peptide 16 plasma rapidly increased, then within 6 hours after injection infusion remained quantifiable in animals that were administered the peptide 16/lipid complex at doses of 15 and 30 mg/kg Detected levels of peptide 16 was observed up to 12 hours in animals of both sexes treated with 60 mg/kg As expected, in the case of intravenous Tmaxwas achieved immediately after injection. It was found that the half-life of circulating levels of peptide 16 is from 0.5 to 5 hours in rats of both sexes, and it increases with increasing input dose. Clearance and volume of distribution decreased with increasing input dose. Volume-based distribution, it was possible to conclude that the peptide 16/lipid complex was generally distributed throughout the volume of plasma.

Example 17

Measurement of the pharmacokinetics of the peptide 16/lipid complex in monkeys

The pharmacokinetics of the peptide 16/lipid complex (where the lipids are represented by sphingomyelin, DPPC, and DPPG in a weight ratio of 1:1,2125:1,2125:of 0.075, and the peptide: lipid weight ratio is 1:2,5) were evaluated in macaques-Griboedov.

Animals in the control group, the media was injected intravenously 10 ml/kg 130-millimolar sodium chloride, 12 mm phosphate buffer, pH of 8.2. Animals in groups treated peptide 16/lipid complex, introduced it by 15, 30 or 60 mg/kg every other day the way of the intravenous infusion. The blood was collected in tubes containing Na3EDTA as anticoagulant, at the starting point, at the end of infusione, and then after 1, 2, 6, 12, 24 and 48 hours after administration of the dose. At each time point, approximately 1 ml of blood was collected from the femoral vessel, the animals were fixed and treated without anestesiologia. Plasma was separated by subsequent centrifugation and frozen at -20°C until the analysis. The levels of peptide 16 was analyzed by the method LC-MS/MS as described in example 8. Pharmacokinetic parameters of individual plasma concentrations were determined by pharmacokinetic analysis without the compartmentalization using kinetics 4.4.1. Peptide 16 was determined in plasma after 12 hours after the end of infusion in rats of both sexes, who have introduced the peptide 16/lipid complex at a dose of 15 mg/kg Detected levels of peptide 16 was observed up to 24 hours in animals treated with 30 and 60 mg/kg Levels of phospholipids was also increased after administration of the dose, and then returned to the original levels over periods of time comparable with those for peptide 16. As expected, in the case of intravenous Tmaxwas achieved immediately after a dose. It was found that the half-life of circulating levels of peptide 16 is from 2 to 7 hours in monkeys of both sexes, and what about the increases with increasing input dose. Clearance and volume of distribution decreased with increasing input dose. Volume-based distribution, it was possible to conclude that the peptide 16/lipid complex normally distributed primarily in the plasma compartment.

Example 18

Mobilization of cholesterol in rabbits

a. Obtaining the peptide 16/lipid complex

Peptide 16 was synthesized by F-moc synthesis in accordance with example 3 and purified by obremenitve chromatography as described in example 4.

Then the peptide 16 was formed complex in a mixture with sphingomyelin, DPPG and DPPC in the joint lyophilization of solutions of peptide 16, sphingomyelin, DPPG and DPPC in a mixture of glacial acetic acid: water. The resulting dried powder was restored buffer (sodium phosphate buffer, 12 mm, pH 8,2) forming a suspension of the peptide 16/lipid complex having a weight ratio of peptide 16:sphingomyelin:DPPC:DPPG, comprising 1:1,35:1,35:0,30.

b. The introduction of a peptide 16/lipid complex rabbits

New Zealand rabbits (males) weighing 3 to 4 kg, were used in order to demonstrate the mobilization of cholesterol peptide 16/lipid complex.

Conditions in the room, which contained animals, were as follows: temperature, 22±2°C; relative humidity, 55+15%; and 12-hour light cycle is/12-hour dark cycle.

The animals were acclimatized for at least 7 days before the beginning of the research. The animals were housed on a daily unlimited controlled briketirovannogo diet. Unlimited access to water animals had on the course of the study.

Before the introduction of the peptide 16/lipid complex animals during the night were deprived of access to food. Animals were weighed immediately prior to introduction of the peptide 16/lipid complex. The peptide 16/lipid complex was injected intravenously at a dose of 20 mg/kg Injected volume was determined according to the weight. Feeding was resumed after approximately 6 hours after administration of the peptide 16/lipid complex.

c. Analysis of blood samples

Before collecting blood samples of the animals during the night deprived of access to food. The blood was collected at baseline, then after 5 min, 15 min, 30 min, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 30 hours and 34 hours after the start of infusion. Blood samples were collected from the jugular vein or from the marginal ear vein. Blood was collected from the jugular vein by syringe equipped with a needle with EDTA (approximately 1 ml of blood during the selection of the sample). Immediately after blood collection, the blood samples were stored at approximately 4°C, to avoid changes in blood samples. The blood samples were centrifuged (3500 g for 10 minutes at approximately 5°C). Education is subjected to plasma was separated, was taken from them aliquots (3 aliquots at least a volume of 200 μl (aliquots A, B, C)) and stored at approximately -80°C. the Remaining blood clot was thrown.

Serum phospholipids (Phospholipid B, Set # 990-54009, Wako Chemicals GmbH, Neuss, Germany), triglycerides (Triglycerides, Set # 1488872, Boehringer Mannheim Corporation, Indianapolis, Ind.), total cholesterol and neeterificirovannah cholesterol were determined using commercially available kits for automatic analyzer Hitachi 912 (Roche Diagnostics Corporation, Indianapolis, Ind.).

The lipoprotein profiles were analyzed using gel-filtration chromatography on a column (1×30 cm sepharose 6HR, equipped with a detector total or free cholesterol in the on-line mode, as described Kieft et al. (J Lipid Res 1991; 32: 859-866, 1991). The area under the peaks corresponding to the lipoproteins of the size of VLDL, LDL and HDL, integrated. The fraction of free or total cholesterol of each peak was multiplied by the total cholesterol in plasma free cholesterol, which is determined by an automatic analyzer for the determination of VLDL, LDL and HDL of free and total cholesterol. Esterified cholesterol in serum and in the lipoprotein fractions VLDL, LDL and HDL was calculated by subtracting the indices of free cholesterol from the level of total cholesterol.

Increasing the fraction of HDL to total cholesterol after which pusii complexes was plotted as a function of time, and this is illustrated in Fig.6. Total HDL cholesterol increased with the introduction of the peptide 16/lipid complex, which indicates that tissue mobilization of cholesterol and a shift towards the paps.

Example 19

Mobilization of cholesterol in rabbits

a. Obtaining the peptide 16/lipid complex

The peptide 16/lipid complex was obtained in accordance with example 12. The peptide 16/lipid complex had a weight ratio of peptide 16:sphingomyelin:DPPC:DPPG of 1:1,2125:1,2125:0,075 and the weight ratio of peptide:lipid of 1:2,5.

b. The introduction of a peptide 16/lipid complex rabbits

New Zealand rabbits (males) weighing 3 to 4 kg, were used in order to demonstrate increasing levels of HDL in rabbits under the action of the peptide 16/lipid complex.

Conditions in the room, which contained animals, were as follows: temperature, 22±2°C; relative humidity, 55+15%; and 12-hour cycles of light/12-hour dark cycle.

The animals were acclimatized for at least 7 days before the beginning of the research. The animals were housed in an unlimited daily controlled briketirovannogo diet. Unlimited access to water animals had throughout the study.

Before the introduction of the peptide 16/lipid complex animals during the night were deprived of access is to food. Animals were weighed immediately prior to introduction of the peptide 16/lipid complex. To study the minimum dose, which may be registered mobilization of cholesterol, animals were injected with 2.5, 5, and 10 mg/kg of the peptide 16/lipid complex or phosphate buffered saline in the control. To study each dose was taken on four animals per group. Feeding was resumed after approximately 6 hours after administration of the peptide 16/lipid complex or phosphate buffered saline in the control.

c. Analysis of blood samples

Before collecting blood samples of the animals during the night deprived of access to food. The blood was collected at baseline, then after 5 min, 15 min, 30 min, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 30 hours and 34 hours after the start of infusion. Blood samples were collected from the jugular vein or from the marginal ear vein. Blood was collected from the jugular vein by syringe equipped with a needle with EDTA (approximately 1 ml of blood during the selection of the sample). Immediately after blood collection, the blood samples were stored at approximately 4°C, to avoid changes in blood samples. The blood samples were centrifuged (3500 g for 10 minutes at approximately 5°C). Plasma samples were separated, was taken from them aliquots (3 aliquots of at least 200 ál (aliquots A, B, C)) and stored at approximately-80°C. The remaining blood clot was thrown.

Serum phospholipids (Phospholipid B, Set # 990-54009, Wako Chemicals GmbH, Neuss, Germany), triglycerides (Triglycerides, Set # 1488872, Boehringer Mannheim Corporation, Indianapolis, Ind.), total cholesterol and neeterificirovannah cholesterol were determined using commercially available kits for automatic analyzer Hitachi 912 (Roche Diagnostics Corporation, Indianapolis, Ind.).

The lipoprotein profiles were analyzed using gel-filtration chromatography on a column (1×30 cm sepharose 6HR, equipped with a detector total or free cholesterol in the on-line mode, as described Kieft et al. (J Lipid Res 1991; 32: 859-866, 1991). The area under the peaks corresponding to the lipoproteins of the size of VLDL, LDL and HDL, integrated. The fraction of free or total cholesterol of each peak was multiplied by the total cholesterol in plasma free cholesterol, which is determined by an automatic analyzer for the determination of VLDL, LDL and HDL of free and total cholesterol. Esterified cholesterol in serum and in the lipoprotein fractions VLDL, LDL and HDL was calculated by subtracting the indices of free cholesterol from the level of total cholesterol.

Increasing the fraction of HDL free cholesterol after infusion of the complexes was plotted as a function of time, and this is illustrated in Fig.7. A clear increase in the fraction LPF the free cholesterol in the background baseline was observed at a dose of 2.5 mg/kg, indicating the high potential of the peptide 16/lipid complex. Five and 20 minutes after the start of infusions cholesterol was increased by 30% over baseline levels. This increase was statistically significant (p<0,05, according to the coupled deparametrization T-student test). In contrast, no changes from baseline in the placebo group were not found.

The pharmacological effect of the peptide 16/lipid complex at a dose of 2.5 mg/kg was further confirmed by comparing the original scans of lipoprotein fractions, eluruumiks with a column by size exclusion HPLC as illustrated Fig. On the HPLC chromatograms after injection there is a clear increase in the fraction of HDL free cholesterol relative to the base level.

Example 20

The dose-response of the peptide 16/lipid complex

The dose-response of the peptide 16/lipid complex (where the lipids are represented by sphingomyelin, DPPC, and DPPG in a weight ratio of 1:1,2125:1,2125:of 0.075, and the peptide: lipid weight ratio is 1:2,5) was evaluated in new Zealand white rabbits.

Necormancer new Zealand white rabbits used as a model for the mobilization of cholesterol, the peptide 16/lipid complex at concentrations of 5, 10 or 15 mg/ml (based on the concentration of peptide) or phosphate buffered saline in quality is TBE control of the media, was injected intravenously at a rate of 1 ml/min necormancer animals with volume infusion of 2 ml/kg In each group used three animals per dose. The final doses were 0, 10, 20 or 30 mg/kg of Blood was collected at baseline, then after 5 min, 15 min, 30 min, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 30 hours and 34 hours after the start of infusion. Then determined the levels of lipid and lipoprotein in plasma. Lipoprotein levels were determined using exclusion by size fractionation HPLC, with built-in detector of free and total cholesterol, in accordance with the method described Usui, S., Hara, Y., Hosaki, S., and Okazaki, M., A new on-line dual enzymatic method for simultaneous quantification of cholesterol and triglycerides in lipoproteins by HPLC. J. Lipid Res. 43, 805-814 (2002). The area under the peaks corresponding to the lipoproteins of the size of VLDL, LDL and HDL, integrated. The fraction of free or total cholesterol in each peak was multiplied by the level of total cholesterol in plasma free cholesterol to determine the level of cholesterol in each fraction. Levels of ester cholesterol in each fraction was determined by subtracting the indices of free cholesterol from the level of total cholesterol in each fraction. In this model, increased levels of HDL cholesterol in plasma is an indicator of tissue mobilization of cholesterol and bias in Starovoit.

Figure 9 shows the dose-dependent increase of phospholipids in the plasma after infusion rabbits with the peptide 16/lipid complex. This increase reflects the levels of circulating peptide 16/lipid complex, as the phospholipid is a component of the peptide 16/lipid complex. The levels of peptide 16 was formed peak within the first 30 minutes, and then declined to baseline levels. There was also a dose-dependent increase in the mobilization of cholesterol. This followed from the fact that increased as the levels of total cholesterol in plasma (figa)and levels of total cholesterol, HDL cholesterol (figa). The main part of high cholesterol was presented in the form of free cholesterol (figure 10 and 11).

An increase in total and free cholesterol in the LDL fraction (figs and 11D) was observed at the two highest doses. Increased levels of free cholesterol was approximately equal to that of total cholesterol, indicating a small increase of ester cholesterol in the specified faction. The increase in free cholesterol in the LDL fraction, in the absence of increase of ester cholesterol, indicates that this increase may not reflect the increase in the level typical LDL, rich in complex ester of cholesterol. The complexes appearing in this fraction lipoprotein, most likely are the product of infusion the th peptide 16/lipid complex which caused the increase of cholesterol in the process of mobilization of cholesterol. The observed increase of VLDL cholesterol was distributed between esterified and neeterificirovannah fractions of cholesterol. The levels of triglycerides transit increased during the first four to six hours after administration of all doses of the peptide 16/lipid complex (Fig). Not found obvious relationship between the introduced dose and increased levels of triglycerides.

Example 21

The minimum effective dose of the peptide 16/lipid complex

The minimum effective dose of the peptide 16/lipid complex (where the lipids are represented by sphingomyelin, DPPC, and DPPG in a weight ratio of 1:1,2125:1,2125:of 0.075, and the peptide: lipid weight ratio is 1:2,5) was evaluated in new Zealand white rabbits.

Studied the minimum dose at which you can register the mobilization of cholesterol. Animals were treated with 0, 2.5, 5, and 10 mg/kg of the peptide 16/lipid complex. To study each dose was taken on four animals per group. Pharmacological effect was most evident when the level of free cholesterol in the HDL fraction compared with baseline levels (Fig). This was to be expected, since most elevated cholesterol levels after infusion of a peptide 16/lipid complex, presents a free holes what Erin in the HDL fraction. In addition, free cholesterol represented approximately one third of total HDL cholesterol, which facilitates the detection of increased levels of this fraction. There has been a clear increase in free HDL cholesterol above the base level at the dose of 2.5 mg/kg five and 20 minutes after the start of infusions cholesterol was increased by 30% over baseline. This increase was statistically significant (p<0,05, according to the coupled deparametrization T-student test). In contrast, no changes from baseline in the control group were not found.

The pharmacological effect of the peptide 16/lipid complex at a dose of 2.5 mg/kg or 5 mg/kg was further confirmed by comparing the original scans of lipoprotein fractions, eluruumiks with a column by size exclusion HPLC. As you can see in these two examples, Fig, on the HPLC chromatograms is a clear increase in free cholesterol relative to the base level in the HDL fraction. This is shown by the increase in the area under the peak for HDL sample taken 20 minutes after the start of infusion of a peptide 16/lipid complex (the bright line on Fig), compared with the area under the peak of HDL at a basic level (the dark line on Fig).

Example 22

The effect of the rate of infusion on the effectiveness of the peptide 16/lipid sets the KSA

The effect of the rate of infusion on the effectiveness of the peptide 16/lipid complex (where the lipids are represented by sphingomyelin, DPPC, and DPPG in a weight ratio of 1:1,2125:1,2125:of 0.075, and the peptide:lipid weight ratio is 1:2,5) was evaluated in new Zealand white rabbits.

Studied the effect of the rate of infusion of a peptide 16/lipid complex on the mobilization of cholesterol. The peptide 16/lipid complex at a concentration of 10 mg/ml (based on the concentration of peptide) or phosphate buffered saline as a control medium were injected in a volume dose of 2 ml/kg at a rate of either 1 ml/min, or 0.2 ml/min Final dose of the peptide 16/lipid complex was 20 mg/kg four animals were studied in groups of a peptide 16/lipid complex and two animals in the control groups of the carrier. The weight of the rabbits ranged from 2.2 to 2.8 kg

The rate of increase of the level of phospholipid in the plasma as a result of infusion of a peptide 16/lipid complex and the rate of increase of level of cholesterol in the plasma as a result of subsequent mobilization of cholesterol was slower in animals, which the peptide 16/lipid complex was injected at a slower rate. However, the maximum levels of mobilization of phospholipid and cholesterol were similar. On Fig shows that the increase in free HDL cholesterol after infusion of a peptide 16/lipid complex at a rate of 1 ml/is in or 0.2 ml/min was similar. Thus, in this model the rate of infusion of a peptide 16/lipid complex is greater than the test speed, a small effect or no effect on the mobilization of cholesterol.

Example 23

Pharmacokinetic studies of the peptide 16/lipid complex in rats and monkeys

The pharmacokinetics of the peptide 16/lipid complex (where the lipids are represented by sphingomyelin, DPPC, and DPPG in a weight ratio of 1:1,2125:1,2125:of 0.075, and the peptide: lipid weight ratio is 1:2,5) has been evaluated in rats and monkeys.

a. Research methodology

The concentration of the peptide 16/lipid complex in the plasma of rats and monkeys were determined using liquid chromatography using a hardware tandem mass spectrometry (LC-MS/MS). Peptide 16, a component of the peptide 16/lipid complex were extracted from plasma containing EDTA, after precipitation of the protein fraction acetonitrile. This method allows the measurement of extracted peptide 16 and the internal standard isotope-labeled peptide 16. Extracts resuspendable, and the peptide was studied using ultra-efficient liquid chromatography coupled with tandem mass spectrometer (MS/MS). Calibration interval specified method is 1-500 μg/ml for sample volume of 25 µl. Methods of extraction and LC-MS/MS were based on the General recommendations about what to Westley bioanalytical method of establishing validity and in accordance with the instructions GLP (Good Laboratory Practice). The validation data showed that these methods are sufficiently sensitive, specific, reliable and accurate for determining the peptide 16/lipid complex in the plasma of rats and monkeys.

b. Pharmacokinetic studies in rats

9 rats of each sex per group were included to assess toxicokinetic peptide 16/lipid complex after the introduction of dosing on day 0 (first dose) and day 26. Animals in the control group media received intravenous 20 ml/kg 130-millimolar sodium chloride, 12 mm phosphate buffer, pH of 8.2. Animals in groups treated peptide 16/lipid complex, introduced it by 15, 30 or 60 mg/kg every other day by intravenous infusion. Approximately 0.5 ml of blood was collected from retroorbital sinus under isoflurane anesthesia and collected in tubes containing Na3EDTA as anticoagulant, cohort 3 animals per group at the starting point and through 0,0833, 0,5, 1, 2, 6, 12, 24 and 48 hours after the dose on day 0 and day 26. Thus, each cohort of animals, the blood was collected at three different time points. Plasma was separated by subsequent centrifugation and frozen at -20°C for a period of up to analysis [on the set] Covance (UK). The levels of the peptide was analyzed by the method LC-MS/MS. Toxicokinetics parameters were determined from the individual who's concentrations in plasma by pharmacokinetic analysis without the compartmentalization using kinetics 4.4.1.

As shown in Fig and 17, the levels of peptide 16 in plasma after administration of the dosage rapidly increased, then within 6 hours after injection infusion remained quantifiable in animals that were administered the peptide 16/lipid complex at doses of 15 and 30 mg/kg Detected levels of the peptide was observed up to 12 hours in animals of both sexes treated with a dose of 60 mg/kg Levels of phospholipids was increased after administration of the dose, and then returned to the original levels at time intervals similar to those for the indicated peptide. The level of free cholesterol (neeterificirovannah) was increased after infusion dose-dependent manner, which is an indicator of mobilization of cholesterol. After that, there was a decrease of cholesterol, suggesting that the particles of the peptide 16/lipid complex effectively remove cholesterol from circulation. A similar picture was observed on day 0 and day 26.

Average values toxicokinetics parameters for the peptide 16/lipid complex on day 0 (first dose) and day 26 (last dose) are presented below in table 11:

Table 11
Toxicokinetics parameters of the peptide 16/lipid complex in rats
Day Dose (mg/kg)FloorCmax(ug/kg)Tmax(h)AUC0-12 (µg·h/ml)T½ (h)CL (ml/kg/h)Vd (ml/kg)
Day 015Males3410,08338511,3618,035,4
Day 030Males6630,083322911,2813,124,1
Day 060Males13900,083374972,16of 7.9024,7
Day 015Female2870,0833671 0,83522,527,1
Day 030Female6880,083321061,3514,628,3
Day 060Female14270,083366891,728,9322,1
Day 2615Males4220,083311761,7113,032,1
Day 2630Males8580,083331881,629,37of 21.9
Day 2660Males1870 1,0098892,565,8621,6
Day 2615Female3860,08338411,0118,126,3

Day 2630Female8150,083324901,4112,325,1
Day 2660Female15370,083378041,79of 7.6419,7
The parameters calculated from the levels of peptide 16 plasma dynamics in time.

Tmaxwas achieved immediately after a dose. Set the half-life of circulating levels of peptide 16 was 0,835 to 2.56 hours in both male and female rats in the and, and it increased with the increase of the input dose. Clearance and volume of distribution decreased with increasing input dose. Volume-based distribution, it was possible to conclude that the peptide 16/lipid complex normally distributed primarily in the plasma compartment (reference volume of plasma in rats = 30 ml/kg). Cm. Davies, B and Morris, T. Physiological parameters in laboratory animals and human, Pharmaceutical Research, 10, 1093-1095, 1993.

The increase in AUC(0-12 h)and Cmaxwith increasing doses (based on a dose of 15 mg/kg) are presented in table 12. Values of Cmaxwere proportional to the dose in both sexes, whereas the values of AUC(0-12 h)increased more than proportionally with dose, indicating a large retention time of the peptide 16/lipid complex in the circulation with increasing dose.

Table 12
Increased AUC and Cmaxwith increasing doses of the peptide 16/ lipid complex
Dose
15 mg/kg30 mg/kg60 mg/kg
MalesFemalesMales FemalesMalesFemales
Day 0
Increment dose112244
The increase in AUC(0-12 h)--2,693,148,819,96
The increase in Cmax--1,942,44,07to 4.98
Day 26
Increment dose112244
The increase in AUC(0-12 h)--2,712,968,4 9.28 are
The increase in Cmax--2,032,114,433,98

There were no significant associated with sex differences in pharmacokinetic profiles, the values of AUC or Cmaxafter administration of single dose and multiple doses. On the basis of indicators Cmaxand AUC, during the 4-week period of introduction failed to detect accumulation of the peptide 16 or peptide 16/lipid complex.

c. Pharmacokinetic studies in monkeys

The toxicokinetics of the peptide 16/lipid complex was evaluated after administration of doses on day 0 (first dose) and day 26. Animals in the control group media received intravenously 10 ml/kg 130-millimolar sodium chloride, 12 mm phosphate buffer, pH of 8.2. Animals in groups treated peptide 16/lipid complex, introduced it by 15, 30 or 60 mg/kg every other day by intravenous infusion. The blood was collected in tubes containing Na3EDTA as anticoagulant, at the starting point, at the end of infusione, and then after 1, 2, 6, 12, 24 and 48 hours after the dose on day 0 and day 26. At each time point, approximately 1 ml of blood was collected from the femoral vessel, the animals were fixed is, and without anestesiologia. Plasma was separated after centrifugation and frozen at -20°C for a period of up to analysis [on the set] Covance (UK). The levels of the peptide was analyzed by the method LC-MS/MS. Toxicokinetics parameters were determined from the individual concentrations in plasma by pharmacokinetic analysis without the compartmentalization using kinetics 4.4.1.

As shown in Fig and 19, the levels of peptide 16 in the plasma could be detected up to 12 hours after the last infusion in animals of both sexes treated with a dose of the peptide 16/lipid complex in the 15 mg/kg Detected levels of the peptide was observed up to 24 hours in animals treated with 30 and 60 mg/kg Levels of phospholipids was also increased after administration of the dose, and then returned to the original levels at time intervals similar to those for the indicated peptide. The level of free cholesterol (neeterificirovannah) was increased after infusion dose-dependent manner, which is an indicator of mobilization of cholesterol. After that, there was a decrease of cholesterol, suggesting that the particles of the peptide 16/lipid complex effectively remove cholesterol from circulation. A similar picture was observed on day 0 and day 26.

Average values toxicokinetics parameters for the peptide 16/lipid complex h is day 0 (first dose) and day 26 (last dose) are presented below in table 13:

Table 13
Toxicokinetics parameters of the peptide 16/lipid complex in monkeys
DayDose (mg/kg)FloorCmax(ug/kg)Tmax(h)AUC0-12 (µg·h/ml)T½ (h)CL (ml/kg/h)Vd (ml/kg)
Day 015Males3410,016713462,4211,5039,6
Day 030Males735043372,966,9029,3
Day 060Males15400137874,58 4,2728,1
Day 015Female365013832,3711,4to 38.3
Day 030Female736043373.04 fromfor 6.8129,4
Day 060Female15080131683,244,5421,1
Day 2615Males433015392,6610,0038,8
Day 2630Males8240 38902,198,5826,3
Day 2660Males16740121822,825,0720,8
Day 2615Female408014372,1110,9032,8
Day 2630Female690034162,50cent to 8.8532,0
Day 2660Female16080135963,634,5122,9
The parameters calculated from the levels of peptide 16 plasma in dyn is ke in time.
T=0 corresponds to the end of infusions.

Tmaxwas achieved immediately after a dose. Set the half-life of circulating levels of peptide 16 was of 2.11 to 4.58 hours in rats of both sexes, and it was increased dose-dependently. Clearance and volume of distribution decreased with increasing input dose. Volume-based distribution, it was possible to conclude that the peptide 16/ lipid complex normally distributed primarily in the plasma compartment (reference volume of plasma in rats = 45 ml/kg). Cm. Davies, B and Morris, T. Physiological parameters in laboratory animals and human, Pharmaceutical Research, 10, 1093-1095, 1993.

The increase in AUC(0-24 h)and Cmaxwith increasing doses (based on a dose of 15 mg/kg) are presented in table 14. Values of Cmaxwere proportional to the dose in both sexes, whereas the values of AUC(0-24 h)increased more than proportionally with dose, indicating a large retention time of the peptide 16/lipid complex in the circulation with increasing dose.

Table 14
Increased AUC and Cmaxwith increasing doses of the peptide 16/lipid complex
Dose
15 mg/kg 30 mg/kg60 mg/kg
MalesFemalesMalesFemalesMalesFemales
Day 0
Increment dose112244
The increase in AUC(0-24 h)--3,223,3110,29,52
The increase in Cmax--2,152,014,514,13
Day 26
Increment dose112244
The increase in AUC(0-24 h)--2,532,38to $ 7.919,46
The increase in Cmax--to 1.861,683,783,94

There were no significant associated with sex differences in pharmacokinetic profiles, the values of AUC or Cmaxafter administration of single dose and multiple doses.

On the basis of indicators Cmaxand AUC, during the 4-week period of introduction failed to detect accumulation of the peptide 16 or peptide 16/lipid complex.

Example 24

Pharmacokinetic studies of peptide 16 and the peptide 16/lipid complexes in mice

After injection of one of three peptide compositions in plasma measured total cholesterol, neeterificirovannah cholesterol and ester cholesterol (as the difference between total and neeterificirovannah cholesterol).

Peptide compositions: (A) the peptide 16; (B) a complex of a peptide 16/DPPC (weight ratio 1:2); (C) a complex of a peptide 16/DPPC (weight ratio 1:2,5). Each of compositions A, B and C was obtained in the form of solutions in to the concentrations of 15 mg/ml

20 mice C57B1/6J restricted power in at least two weeks, in accordance with a prescribed diet for rodents, containing 60% kcal% fat (Reseach diets - D12492). In drinking water were added 5% glucose. After 3 hours of fasting they were introduced peptide composition at a dose of 30 mg/kg (intravenous injection of 50 ál), after which blood samples were taken after 15, 30, 60, 120 and 240 minutes. One blood sample was taken before injection.

Plasma samples were analyzed to determine total cholesterol (sets of production Biolabo-CEROOX-SOP002, CER00X-SOP003). Ester cholesterol was calculated as the difference between total cholesterol and neeterificirovannah cholesterol.

The results presented in Fig and 22.

In this application provides a number of links, each of which is fully incorporated into this description by reference.

This is followed by some illustrative embodiments of the present invention:

1. A peptide containing from 22 to 29 residues having the following formula I:

R1-Y1-X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-X18-X19-X20-X21-X22-X23-Y2-R2

The formula I

or its pharmaceutically acceptable salt, where

X 1is missing or achiral amino acid residue, D-basic amino acid residue or L-basic amino acid residue;

X2is achiral amino acid residue, D-basic amino acid residue or L-basic amino acid residue;

X3is aliphatic achiral amino acid residue, an aliphatic D-amino acid residue or an aliphatic L-amino acid residue;

X4is achiral basic amino acid residue, D-basic amino acid residue or L-basic amino acid residue;

X5represents Gln, Asn, D-Gln, D-Asn or achiral basic amino acid residue, D-basic amino acid residue or L-basic amino acid residue;

X6is achiral basic amino acid residue, D-basic amino acid residue or L-basic amino acid residue;

X7is achiral hydrophobic amino acid residue, D-hydrophobic amino acid residue is L or a hydrophobic amino acid residue;

X8is achiral hydrophobic amino acid residue, D-hydrophobic amino acid residue is L or a hydrophobic amino acid residue;

X9is achiral hydrophilic amino acid residue, D-hydrophilic amino acid residue is whether L is a hydrophilic amino acid residue;

X10represents Leu, Trp, Gly, Nal, D-Leu, D-Trp or D-Nal;

X11represents Gly or achiral aliphatic amino acid residue, D-aliphatic amino acid residue or L-aliphatic amino acid residue;

X12is achiral hydrophilic amino acid residue, D-hydrophilic amino acid residue or L-hydrophilic amino acid residue;

X13is achiral hydrophilic amino acid residue, D-hydrophilic amino acid residue or L-hydrophilic amino acid residue;

X14represents Leu, Trp, Gly, D-Leu, or D-Trp;

X15represents Leu, Gly or D-Leu;

X16is achiral acidic amino acid residue, D-acidic amino acid residue or L-acidic amino acid residue;

X17is achiral hydrophilic amino acid residue, D-hydrophilic amino acid residue or L-hydrophilic amino acid residue;

X18represents Leu, Phe, D-Leu or D-Phe;

X19represents Leu, Phe, D-Leu or D-Phe;

X20is achiral acidic amino acid residue, D-acidic amino acid residue or L-acidic amino acid residue;

X21represents Leu, Phe, D-Leu or D-Phe;

X22is achiral aliphatic amino acid residue, D-alifatic the sky amino acid residue or L-aliphatic amino acid residue; and

X23represents Inp, Nip, azPro, Pip, azPip, D-Nip or D-Pip;

Y1is absent or is an amino acid sequence containing from 1 to 7 residues, each residue of the sequence is independently achiral, D - or L-amino acid residue;

Y2is absent or is a sequence containing from 1 to 7 amino acid residues, each residue of the sequence is independently achiral, D - or L-amino acid residue;

R1represents H or aminosidine group; and

R2represents OH or a carboxyl protective group; and

a) all amino acid residues that are not terminal amino acid residues and residues directly adjacent to the terminal amino acid residues are achiral or L-amino acid residues; or

b) all amino acid residues that are not terminal amino acid residues and residues directly adjacent to the terminal amino acid residues are achiral or D-amino acid residues.

2. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 1, where

X3represents Leu or D-Leu;

X7represents Leu, Gly, Nal, D-Leu or D-Nal;

X8represents Ala, Nal, Trp, Gly, Leu, Phe, D-Aa, D-Nal, D-Trp, D-Leu or D-Phe;

X11represents Leu, Gly, Aib or D-Leu; and

X22represents Ala, Leu, Val, D-Ala, D-Leu or D-Val.

3. The peptide or pharmaceutically acceptable salt of a peptide according to embodiment 1, where

X1absent or represents Lys or D-Lys;

X2represents Lys, Orn, D-Lys or D-Orn;

X4represents Lys, Orn, D-Lys or D-Orn;

X5represents Gln, Asn, Lys, Orn, D-Gln, D-Asn, D-Lys or D-Orn;

X6represents Gln, Asn, Lys, Orn, D-Gln, D-Asn, D-Lys or D-Orn;

X9represents Asp, Glu, D-Asp or D-Glu;

X12represents Glu, Asp, D-Asp or D-Glu;

X13represents Asn, Gln, D-Asn or D-Gln;

X16represents Asp, Glu, D-Asp or D-Glu;

X17represents Lys, Arg, Orn, D-Lys, D-Arg or D-Orn; and

X20represents Asp, Glu, D-Asp or D-Glu.

4. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 3, where X18represents Phe or D-Phe.

5. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 1, where the specified peptide is a peptide containing 22 or 23 of the peptide residue.

6. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 5, where R1represents H and R2represents OH.

7. The peptide or pharmaceutically acceptable Sol is specified peptide, according to the embodiment 5, where

X1missing, represents Lys or D-Lys;

X2represents Lys, Orn, D-Lys or D-Orn;

X3represents Leu or D-Leu;

X4represents Lys, Orn, D-Lys or D-Orn;

X5represents Gln, Asn, Lys, Orn, D-Gln, D-Asn, D-Lys or D-Orn;

X6represents Lys, Orn, D-Lys or D-Orn;

X7represents Gly, Leu, Nal, D-Leu or D-Nal;

X8represents Ala, Nal, Trp, Leu, Phe, Gly, D-Ala, D-Nal, D-Trp, D-Leu or D-Phe;

X9represents Asp, Glu, D-Asp or D-Glu;

X11represents Gly, Leu, Aib or D-Leu;

X12represents Glu, Asp, D-Glu or D-Asp;

X13represents Asn, Gln, D-Asn or D-Gln;

X16represents Asp, Glu, D-Asp or D-Glu;

X17represents Lys, Arg, Orn, D-Lys, D-Arg or D-Orn;

X20represents Asp, Glu, D-Asp or D-Glu; and

X22represents Ala, Val, Leu, D-Ala, D-Val or D-Leu.

8. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 7, where X5represents Gln, Asn, D-Gln or D-Asn, X6represents Lys, Orn, D-Lys or D-Orn; or X5represents Lys, Orn, D-Lys or D-Orn, and X6represents Gln, Asn, D-Gln or D-Asn.

9. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 7, where X1is absent, and the peptide is a peptide containing 22 is peptidnyh balance. D

10. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 9, where each of X7X8X10X11X14and X15is other than Gly.

11. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 9, where only one of X7X8X10X11X14and X15is Gly.

12. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 9, where

X2and X4represents Lys, Orn, D-Lys or D-Orn;

X5represents Gln, Lys, D-Gln or D-Lys;

X9is achiral acidic amino acid residue, D-acidic amino acid residue or L-acidic amino acid residue;

X12represents Glu, Asn, GIn, Arg, D-Glu, D-Asn, D-Gln or D-Arg;

X13represents Glu, Asn, Gln, Arg, D-Glu, D-Asn, D-Gln or D-Arg;

X16is achiral acidic amino acid residue, D-acidic amino acid residue or L-acidic amino acid residue;

X17represents Arg, Lys, Orn, D-Arg, D-Lys or D-Orn;

X21represents Leu or D-Leu; and

X22represents Ala, Val, Leu, D-Ala, D-Val or D-Leu.

13. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 1, where the peptide is a

Lys-Leu-Lys-Gln-Lys-Leu-Trp-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (EQ ID NO:2);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Gly-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:3);

Lys-Leu-Lys-Gln-Lys-Nal-Ala-Glu-Leu-Gly-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:4);

Lys-Leu-Lys-Gln-Lys-Leu-Trp-Glu-Leu-Gly-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:5);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Gly-Glu-Asn-Trp-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:6);

Orn-Leu-Orn-Gln-Orn-Leu-Ala-Glu-Leu-Gly-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:7);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Gly-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Phe-Asp-Leu-Val-Inp (SEQ ID NO:8);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Gly-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Glu-Leu-Val-Inp (SEQ ID NO:9);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Gly-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:10);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Gly-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:11);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Gly-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:12);

Lys-Leu-Lys-Gln-Lys-Leu-Gly-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:13);

Lys-Leu-Lys-Gln-Lys-Gly-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:14);

Lys-Leu-Lys-Gln-Lys-Leu-Nal-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:15);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:16);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Aib-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:18);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Lys-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:19);

Lys-Leu-Lys-Gln-Lys-Nal-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:20);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Gln-Leu-Leu-Glu-Lys-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:21);

Orn-Leu-Orn-Gln-Orn-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:22);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Trp-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:23);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Leu-Leu-Asp-Leu-Val-Inp (SEQ ID NO:24);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Gl-Leu-Leu-Glu-Lys-Phe-Leu-Glu-Leu-Val-Inp (SEQ ID NO:25);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Gln-Leu-Leu-Glu-Lys-Phe-Leu-Glu-Leu-Leu-Inp (SEQ ID NO:26);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Aib-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Phe-Asp-Leu-Val-Inp (SEQ ID NO:28);

Lys-Leu-Lys-Gln-Lys-Leu-Leu-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:29);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Nal-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:30);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Trp-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:31);

Orn-Leu-Orn-Gln-Orn-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Orn-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:32);

Lys-Leu-Lys-Gln-Lys-Leu-Phe-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:33);

Lys-Leu-Lys-Gln-Arg-Leu-Ala-Asp-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Lys-Phe-Leu-Glu-Leu-Val-Inp (SEQ ID NO:36);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Gln-Leu-Leu-Asp-Lys-Phe-Leu-Glu-Leu-Ala-Inp (SEQ ID NO:40);

Lys-Leu-Lys-Gln-Lys-Leu-Trp-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:94);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Gly-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:95);

Lys-Leu-Lys-Gln-Lys-Nal-Ala-Glu-Leu-Gly-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:96);

Lys-Leu-Lys-Gln-Lys-Leu-Trp-Glu-Leu-Gly-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:97);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Gly-Glu-Asn-Trp-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:98);

Orn-Leu-Orn-Gln-Orn-Leu-Ala-Glu-Leu-Gly-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:99);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Gly-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Phe-Asp-Leu-Val-Nip (SEQ ID NO:100);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Gly-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Glu-Leu-Val-Nip (SEQ ID NO:101);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Gly-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:102);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Gly-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:103);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Gly-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:104);

Lys-Leu-Lys-Gln-Lys-Leu-Gly-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:105);

Lys-Leu-Lys-Gln-Lys-Gly-Ala-Glu-Lu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:106);

Lys-Leu-Lys-Gln-Lys-Leu-Nal-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:107);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:108);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Aib-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:110);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Lys-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:111);

Lys-Leu-Lys-Gln-Lys-Nal-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:112);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Gln-Leu-Leu-Glu-Lys-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:113);

Orn-Leu-Orn-Gln-Orn-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:114);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Trp-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:115);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Leu-Leu-Asp-Leu-Val-Nip (SEQ ID NO:116);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Gln-Leu-Leu-Glu-Lys-Phe-Leu-Glu-Leu-Val-Nip (SEQ ID NO:117);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Gln-Leu-Leu-Glu-Lys-Phe-Leu-Glu-Leu-Leu-Nip (SEQ ID NO:118);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Aib-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Phe-Asp-Leu-Val-Nip (SEQ ID NO:120);

Lys-Leu-Lys-Gln-Lys-Leu-Leu-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:121);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Nal-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:122);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Trp-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:123);

Orn-Leu-Orn-Gln-Orn-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Orn-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:124);

Lys-Leu-Lys-Gln-Lys-Leu-Phe-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:125);

Lys-Leu-Lys-Gln-Arg-Leu-Ala-Asp-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Lys-Phe-Leu-Glu-Leu-Val-Nip (SEQ ID NO:128); or

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Gln-Leu-Leu-Asp-Lys-Phe-Leu-Glu-Leu-Ala-Nip (SEQ ID NO:132), or

pharmaceutically acceptable salt of one of the above [connections].

14. The peptide or pharmaceutically acceptable sliptide, according to the embodiment 5, where the specified peptide is a peptide containing 23 peptide residue.

15. The peptide or pharmaceutically acceptable salt of a peptide according to embodiment 14, where the specified peptide is a

Lys-Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:17); or

Lys-Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:109), or

pharmaceutically acceptable salt of one of the above [connections].

16. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 5, where X1is absent, and the peptide is a peptide containing 22 peptide residue.

17. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 16, where the peptide is a

Lys-Leu-Lys-Lys-Gln-Leu-Ala-Glu-Leu-Leu-Glu-Arg-Leu-Leu-Asp-Asn-Phe-Leu-Glu-Leu-Val-Inp (SEQ ID NO:34);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Arg-Leu-Leu-Asp-Asn-Phe-Leu-Glu-Leu-Val-Inp (SEQ ID NO:35);

Lys-Leu-Lys-Lys-Gln-Leu-Ala-Glu-Leu-Leu-Glu-Arg-Leu-Leu-Asp-Asn-Phe-Leu-Glu-Leu-Val-Nip (SEQ ID NO:126); or

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Arg-Leu-Leu-Asp-Asn-Phe-Leu-Glu-Leu-Val-Nip (SEQ ID NO:127), or

pharmaceutically acceptable salt of one of the above [connections].

18. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 16, where

X9represents Gln, Lys, D-Gln, D-Lys, achiral acidic amino acid residue, D-acidic amino acid residue or L-acidic amino acid is hydrated residue;

X12represents Asn, D-Asn, achiral acidic amino acid residue, D-acidic amino acid residue or L-acidic amino acid residue; and

X17represents Asn, Glu, D-Asn, D-Glu, achiral basic amino acid residue, D-basic amino acid residue or L-basic amino acid residue.

19. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 16, where

X9represents Gln, Lys, D-Gln, D-Lys, achiral acidic amino acid residue, D-acidic amino acid residue or L-acidic amino acid residue;

X12represents Asn, D-Asn, achiral acidic amino acid residue, D-acidic amino acid residue or L-acidic amino acid residue; and

X17represents Asn, Glu, D-Asn, D-Glu, achiral basic amino acid residue, D-basic amino acid residue or L-basic amino acid residue.

20. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 16, where

X2represents Lys, Orn, D-Lys or D-Orn;

X3represents Leu or D-Leu;

X4represents Lys, Orn, D-Lys or D-Orn;

X5represents Lys, Orn, GIn, Asn, D-Lys, D-Orn, D-Gln or D-Asn;

X6represents Lys, Orn, D-Lys or D-Orn;

X7represents Leu, Gly, Nal, D-Leu or D-Nal;

X8pre what is Ala, Trp, Gly, Leu, Phe, Nal, D-Ala, D-Trp, D-Leu, D-Phe or D-Nal;

X9represents Asp, Glu, Gln, Lys, D-Asp, D-Glu, D-Gln or D-Lys;

X11represents Leu, Gly, Aib or D-Leu;

X12represents Asp, Glu, Asn, D-Asp, D-Glu or D-Asn;

X13represents Asn, Gln, Glu, Arg, D-Asn, D-Gln, D-Glu or D-Arg;

X16represents Asp, Glu, D-Asp or D-Glu;

X17represents Lys, Arg, Orn, Asn, Glu, D-Lys, D-Arg, D-Orn, D-Asn or D-Glu;

X20represents Asp, Glu, D-Asp or D-Glu; and

X22represents Ala, Val, Leu, D-Ala, D-Val or D-Leu.

21. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 16, where

X9represents Glu or D-Glu;

X12represents Glu or D-Glu;

X13represents Asn, Glu, D-Asn or D-Glu;

X14represents Leu or D-Leu;

X15represents Leu or D-Leu;

X16represents Glu or D-Glu;

X17represents Arg, Lys, D-Arg or D-Lys;

X18represents Phe or D-Phe;

X19represents Leu or D-Leu;

X21represents Leu or D-Leu; and

X22represents Val or D-Val.

22. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 16, where X11represents Gly, and each of X7X8X10X14and X15is other than Gly.

23. The peptide or the pharmacist who Cesky acceptable salt of the indicated peptide, according to the embodiment 16, where

X2represents Lys, Orn, D-Lys or D-Orn;

X3represents Leu or D-Leu;

X4is Lys, Orn, D-Lys or D-Orn;

X5represents Gln or D-Gln;

X6represents Lys, Orn, D-Lys or D-Orn;

X7represents Leu, Nal, D-Leu or D-Nal;

X8represents Ala, Trp, D-Ala or D-Trp;

X9represents Glu or D-Glu;

X10represents Leu or D-Leu;

X11is Gly;

X12represents Glu or D-Glu;

X13represents Asn or D-Asn;

X14represents Leu, Trp, D-Leu, or D-Trp;

X15represents Leu or D-Leu;

X16represents Glu or D-Glu;

X17represents Arg or D-Arg;

X18represents Phe or D-Phe;

X19represents Leu, Phe, D-Leu or D-Phe;

X20represents Asp, Glu, D-Asp or D-Glu;

X21represents Leu or D-Leu; and

X22represents Val or D-Val.

24. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 20, where the specified peptide is a

Lys-Leu-Lys-Gln-Lys-Leu-Trp-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:2);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Gly-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:3);

Lys-Leu-Lys-Gln-Lys-Nal-Ala-Glu-Leu-Gly-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:4);

Lys-Leu-Lys-Gln-Lys-Leu-Trp-Glu-Leu-Gly-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-As-Leu-Val-Inp (SEQ ID NO:5);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Gly-Glu-Asn-Trp-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:6);

Orn-Leu-Orn-Gln-Orn-Leu-Ala-Glu-Leu-Gly-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:7);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Gly-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Phe-Asp-Leu-Val-Inp (SEQ ID NO:8);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Gly-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Glu-Leu-Val-Inp (SEQ ID NO:9);

Lys-Leu-Lys-Gln-Lys-Leu-Trp-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:94);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Gly-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:95);

Lys-Leu-Lys-Gln-Lys-Nal-Ala-Glu-Leu-Gly-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:96);

Lys-Leu-Lys-Gln-Lys-Leu-Trp-Glu-Leu-Gly-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:97);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Gly-Glu-Asn-Trp-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:98);

Orn-Leu-Orn-Gln-Orn-Leu-Ala-Glu-Leu-Gly-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:99);

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Gly-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Phe-Asp-Leu-Val-Nip (SEQ ID NO:100); or

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Gly-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Glu-Leu-Val-Nip (SEQ ID NO:101), or

pharmaceutically acceptable salt of one of the above [connections].

25. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 16, where X15is Gly, and each of X7X8X10X11and X14is other than Gly.

26. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 25, where the specified peptide is a

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Gly-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:10); or

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Gly-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:102), or

pharmaceutically who ramlau salt of one of the above [connections].

27. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 16, where X14is Gly, and each of X7X8X10X11and X15is other than Gly.

28. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 27, where the specified peptide is a

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Gly-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:11); or

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Gly-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:103), or

pharmaceutically acceptable salt of one of the above [connections].

29. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 16, where X10is Gly, and each of X7X8X11X14and X15is other than Gly.

30. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 29, where the specified peptide is a

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Gly-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:12); or

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Gly-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:104), or

pharmaceutically acceptable salt of one of the above [connections].

31. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 16, where X8is Gly, and each of X7X10X11X14and X15is other than Gly.

32. The peptide is whether the pharmaceutically acceptable salt of the indicated peptide, according to embodiment 31, where the specified peptide is a

Lys-Leu-Lys-Gln-Lys-Leu-Gly-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:13); or

Lys-Leu-Lys-Gln-Lys-Leu-Gly-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:105), or

pharmaceutically acceptable salt of one of the above [connections].

33. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 16, where X7is Gly, and each of X8X10X11X14and X15is other than Gly.

34. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 33, where the specified peptide is a

Lys-Leu-Lys-Gln-Lys-Gly-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:14); or

Lys-Leu-Lys-Gln-Lys-Gly-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:106), or

pharmaceutically acceptable salt of one of the above [connections].

35. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 1, where the specified peptide is a

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:16); or

Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:108), or

pharmaceutically acceptable salt of one of the above [connections].

36. A peptide containing from 15 to 22 residues having the following formula II:

R1-Y1-X1-X2-X3-X4-X5-X6 -X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-X18-Y2-R2

Formula II

or its pharmaceutically acceptable salt, where

X1is achiral, D -, or L-basic amino acid residue;

X2represents Leu or D-Leu;

X3is achiral, D -, or L-basic amino acid residue;

X4represents Gln, Asn, D-Gln or D-Asn;

X5represents Leu, D-Leu, or is achiral, D -, or L-basic amino acid residue;

X6represents Leu, Trp, Phe, D-Leu, D-Trp or D-Phe;

X7is achiral, D -, or L-acidic amino acid residue;

X8represents Asn, D-Asn or is achiral, D -, or L-acidic amino acid residue;

X9represents Leu, Trp, D-Leu, or D-Trp;

X10represents Leu, Trp, D-Leu, or D-Trp;

X11is an achiral, D -, or L-acidic amino acid residue;

X12is achiral, D -, or L-basic amino acid residue;

X13represents Leu, Phe, D-Leu or D-Phe;

X14represents Leu, Phe, D-Leu or D-Phe;

X15is achiral, D -, or L-acidic amino acid residue;

X16represents Leu or D-Leu;

X17is achiral, D -, or L-aliphatic amino acid is Tim residue;

X18represents Inp, Nip, azPro, Pip, azPip, D-Nip or D-Pip;

Y1is absent or is an amino acid sequence containing from 1 to 4 residues;

Y2no;

R1represents H or aminosidine group;

R2represents OH or a carboxyl protective group;

and from zero to three residues from the X1to X17absent; and

a) each chiral amino acid residue is L-amino acid residue;

b) each chiral amino acid residue is a D-amino acid residue;

(c) each chiral amino acid residue is L-amino acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is a D-amino acid residue; or

d) each chiral amino acid residue is a D-amino acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is an L-amino acid residue.

37. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 36, where

X17represents Ala, Leu, Val, D-Ala, D-Leu and the and D-Val.

38. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 36, where

X1represents His, Lys, Arg, D-His, D-Lys or D-Arg;

X3represents Lys, Arg, Orn, D-Lys, D-Arg or D-Orn;

X5represents Lys, Arg, Orn, D-Lys, D-Arg or D-Orn;

X7represents Glu or D-Glu;

X8represents Asn, Glu, D-Asn or D-Glu;

X11represents Asp, Glu, D-Asp or D-Glu;

X12represents Arg, Lys, Orn, D-Arg, D-Lys or D-Orn; and

X15represents Asp, Glu, D-Asp or D-Glu.

39. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 38, where

X13represents Phe or D-Phe.

40. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 36, where the specified peptide is a peptide containing 18 residues.

41. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 40, where R1represents H, and R2represents OH.

42. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 41, where

X1represents Arg, Lys, Orn, D-Arg, D-Lys or D-Orn;

X3represents Arg, Lys, Orn, D-Arg, D-Lys or D-Orn;

X5represents Arg, Lys, Orn, D-Arg, D-Lys or D-Orn;

X7represents Glu or D-Glu;

X8represents Glu, Asn, D-Gu or D-Asn;

X11represents Glu, Asp, D-Glu or D-Asp;

X12represents Arg, Lys, Orn, D-Arg, D-Lys or D-Orn;

X15represents Asp, Glu, D-Asp or D-Glu; and

X17represents Ala, VaI, Leu, D-Ala, D-Val or D-Leu.

43. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 36, where the specified peptide is a

Lys-Leu-Lys-Gln-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:53);

Lys-Leu-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO:54);

Lys-Leu-Lys-Gln-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:145); or

Lys-Leu-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip (SEQ ID NO:146), or

pharmaceutically acceptable salt of one of the above [connections].

44. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 36, where the specified peptide is a

H3C(O)C-Lys-Leu-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Leu-Glu-Arg-Leu-Leu-Asp-Leu-Val-Inp-NH2(SEQ ID NO:65);

H3C(O)C-Lys-Leu-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Leu-Glu-Lys-Phe-Leu-Asp-Leu-Val-Inp-NH2(SEQ ID NO:66);

H3C(O)C-Lys-Leu-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp-NH2(SEQ ID NO:67);

H3C(O)C-Lys-Leu-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Leu-Glu-Arg-Phe-Leu-Glu-Leu-Val-Inp-NH2(SEQ ID NO:68);

H3C(O)C-Lys-LeU-Lys-Gln-Lys-LeU-Glu-Glu-Leu-Leu-Glu-Lys-Phe-Leu-Glu-Leu-Val-Inp-NH2(SEQ ID NO:69);

H3C(O)C-Lys-Leu-Lys-Asn-Lys-Leu-Glu-Glu-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp-NH2(SEQ ID NO:70);

H3C(O)C-Lys-Leu-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Leu-Inp-NH2(SEQ ID NO:71);

H3C(O)C-Lys-Le-Lys-Gln-Lys-Leu-Glu-Glu-Trp-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp-NH 2(SEQ ID NO:72);

H3C(O)C-Lys-Leu-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Leu-Glu-Lys-Phe-Leu-Asp-Leu-Leu-Inp-NH2(SEQ ID NO:73);

H3C(O)C-Arg-Leu-Lys-Gln-Arg-Leu-Glu-Glu-Leu-Leu-Asp-Lys-Phe-Leu-Glu-Leu-Ala-Inp-NH2(SEQ ID NO:74);

H3C(O)C-Lys-Leu-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Leu-Glu-Arg-Phe-Phe-Asp-Leu-Val-Inp-NH2(SEQ ID NO:75);

H3C(O)C-Lys-Leu-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Trp-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp-NH2(SEQ ID NO:76);

H3C(O)C-Lys-Leu-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Leu-Glu-Lys-Leu-Leu-Glu-Leu-Leu-Inp-NH2(SEQ ID NO:77);

H3C(O)C-Lys-Leu-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Leu-Glu-Lys-Leu-Leu-Glu-Leu-Val-Inp-NH2(SEQ ID NO:78);

H3C(O)C-Lys-Leu-Arg-Gln-Arg-Leu-Glu-Glu-Leu-Leu-Asp-Lys-Phe-Leu-Glu-Leu-Ala-Inp-NH2(SEQ ID NO:79);

H3C(O)C-Orn-Leu-Orn-Gln-Orn-Leu-Glu-Glu-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp-NH2(SEQ ID NO:80);

H3C(O)C-Lys-Leu-Orn-Gln-Orn-Leu-Glu-Glu-Leu-Leu-Glu-Orn-Phe-Leu-Asp-Leu-Val-Inp-NH2(SEQ ID NO:81);

H3C(O)C-Lys-Leu-Arg-Gln-Arg-Phe-Glu-Glu-Leu-Leu-Asp-Lys-Phe-Leu-Glu-Leu-Ala-Inp-NH2(SEQ ID NO:82);

H3C(O)C-Lys-Leu-Lys-Gln-Lys-Trp-Glu-Glu-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp-NH2(SEQ ID NO:83);

H3C(O)C-Lys-Leu-Lys-Gln-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp-NH2(SEQ ID NO:84);

H3C(O)C-Lys-Leu-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Leu-Glu-Lys-Phe-Leu-Glu-Leu-Leu-Inp-NH2(SEQ ID NO:87);

H3C(O)C-Lys-Leu-Lys-Gln-Glu-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp-NH2(SEQ ID NO:88);

H3C(O)C-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp-NH2(SEQ ID NO:89);

H3C(O)C-Lys-Leu-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Leu-Glu-AIg-Leu-Leu-Asp-Leu-Val-Nip-NH2(SEQ ID NO:157);

H3C(O)C-Lys-Leu-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Leu-Glu-Lys-Phe-Leu-Asp-Leu-Val-Nip-NH2(SEQ ID NO:158);

H3C(O)C-Lys-Leu-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip-NH2

H3C(O)C-Lys-Leu-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Leu-Glu-Arg-Phe-Leu-Glu-Leu-Val-NiP-NH2(SEQ ID NO:160);

H3C(O)C-Lys-Leu-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Leu-Glu-Lys-Phe-Leu-Glu-Leu-Val-NiP-NH2(SEQ ID NO:161);

H3C(O)C-Lys-Leu-Lys-Asn-Lys-Leu-Glu-Glu-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip-NH2(SEQ ID NO:162);

H3C(O)C-Lys-Leu-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Leu-Nip-NH2(SEQ ID NO:163);

H3C(O)C-Lys-Leu-Lys-Gln-Lys-Leu-Glu-Glu-Trp-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip-NH2(SEQ ID NO:164);

H3C(O)C-Lys-Leu-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Leu-Glu-Lys-Phe-Leu-Asp-Leu-Leu-Nip-NH2(SEQ ID NO:165);

H3C(O)C-Arg-Leu-Lys-Gln-Arg-Leu-Glu-Glu-Leu-Leu-Asp-Lys-Phe-Leu-Glu-Leu-Ala-Nip-NH2(SEQ ID NO:166);

H3C(O)C-Lys-Leu-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Leu-Glu-Arg-Phe-Phe-Asp-Leu-Val-Nip-NH2(SEQ ID NO:167);

H3C(O)C-Lys-LeU-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Leu-Glu-Lys-Leu-Leu-Glu-Leu-Leu-Nip-NH2(SEQ ID NO:168);

H3C(O)C-Lys-Leu-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Leu-Glu-LyS-LeU-Leu-Glu-Leu-Val-Nip-NH2(SEQ ID NO:169);

H3C(O)C-Lys-Leu-Arg-Gln-Arg-Leu-Glu-Glu-Leu-Leu-Asp-Lys-Phe-Leu-Glu-Leu-Ala-Nip-NH2(SEQ ID NO:170);

H3C(O)C-Orn-Leu-Orn-Gln-Orn-Leu-Glu-Glu-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip-NH2(SEQ ID NO:171);

H3C(O)C-Lys-Leu-Orn-Gln-Orn-Leu-Glu-Glu-Leu-Leu-Glu-Orn-Phe-Leu-Asp-Leu-Val-Nip-NH2(SEQ ID NO:172);

H3C(O)C-Lys-Leu-Arg-Gln-Arg-Phe-Glu-Glu-Leu-Leu-Asp-Lys-Phe-Leu-Glu-Leu-Ala-Nip-NH2(SEQ ID NO:173);

H3C(O)C-Lys-Leu-Lys-Gln-Lys-Trp-Glu-Glu-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip-NH2(SEQ ID NO:174);

H3C(O)C-Lys-Leu-Lys-Gln-Leu-Leu-Glu-Asn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip-NH2(SEQ ID NO:175);

H3C(O)C-Lys-Leu-Lys-Gln-Lys-Leu-Glu-Glu-Gly-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip-NH2(SEQ ID NO:176);

H3C(O)C-Lys-Leu-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Leu-Glu-Lys-Phe-Leu-Glu-Leu-Leu-Nip-NH 2(SEQ ID NO:179);

H3C(O)C-Lys-Leu-Lys-Gln-Glu-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip-NH2(SEQ ID NO:180); or

H3C(O)C-Lys-Gln-Lys-Leu-Glu-Glu-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Nip-NH2(SEQ ID NO:181), or

pharmaceutically acceptable salt of one of the above [connections].

45. A peptide containing from 22 to 29 residues having the following formula III:

R1-Y1-X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-X18-X19-X20-X21-X22-X23-Y2-R2

Formula III

or its pharmaceutically acceptable salt, where

X1is missing or achiral, D -, or L-basic amino acid residue;

X2is achiral, D -, or L-basic amino acid residue;

X3represents Leu or D-Leu;

X4is achiral, D -, or L-basic amino acid residue;

X5is achiral, D -, or L-basic amino acid residue;

X6represents Gln, Asn, D-Gln or D-Asn;

X7represents Leu or D-Leu;

X8represents Ala or D-Ala;

X9represents Asp or D-Asp;

X10represents Leu, Phe, Gly, D-Leu or D-Phe;

X11represents Gly, Leu or D-Leu;

X12represents Arg or D-Arg;

Xsup> 13is achiral, D -, or L-acidic amino acid residue;

X14represents Leu, Trp, Gly, D-Leu, or D-Trp;

X15represents Leu or D-Leu;

X16represents Gln or D-Gln;

X17represents Glu, Leu, D-Glu or D-Leu;

X18represents Leu, Phe, D-Leu or D-Phe;

X19is achiral, D -, or L-aliphatic amino acid residue;

X20represents Glu or D-Glu;

X21represents Leu, Phe, D-Leu or D-Phe;

X22is achiral, D -, or L-aliphatic amino acid residue;

X23represents Inp, Nip, azPro, Pip, azPip, D-Nip or D-Pip;

Y1is absent or is an amino acid sequence containing from 1 to 7 residues;

Y2is absent or is an amino acid sequence containing from 1 to 7 residues;

R1represents H or aminosidine group;

R2represents OH or a carboxyl protective group;

thus

a) each chiral amino acid residue is L-amino acid residue;

b) each chiral amino acid residue is a D-amino acid residue;

(c) each chiral amino acid residue is L-amino acid residue, except that one or more of each chiral terminal amino acid OST the Cove and each chiral amino acid residue, directly adjacent to them, is a D-amino acid residue; or

d) each chiral amino acid residue is a D-amino acid residue, except that one or more of each chiral terminal amino acid residues and each chiral amino acid residues directly adjacent to them, is an L-amino acid residue.

46. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 45 where the specified peptide is a peptide containing 22 or 23 of the balance.

47. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 46, where the specified peptide is a peptide containing 22 residue.

48. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 47, where X22represents Val, Leu, D-Val or D-Leu.

49. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 47, where

X2represents Lys or D-Lys;

X4represents Lys or D-Lys;

X5represents Lys or D-Lys;

X13represents Glu or D-Glu;

X18represents Phe or D-Phe;

X19represents Leu or D-Leu; and

X22represents Ala, Leu, Val, D-Ala, D-Leu or D-Val.

50. The peptide or pharmaceutically acceptable salt of the specified pepti is a, according to embodiment 47, where

X2represents Lys or D-Lys;

X4represents Lys or D-Lys;

X5represents Lys or D-Lys;

X13represents Glu or D-Glu;

X18represents Phe or D-Phe;

X19represents Leu or D-Leu; and

X22represents Ala, Leu, Val, D-Ala, D-Leu or D-Val.

51. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 47, where X13and X17represents Glu or D-Glu.

52. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 46, where

X1no;

X2represents Lys or D-Lys;

X4represents Lys or D-Lys;

X5represents Lys or D-Lys;

X18represents Phe or D-Phe;

X19represents Leu or D-Leu; and

X22represents Val or D-Val.

53. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 47, where X13or X17represents Glu or D-Glu.

54. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 47, where X22represents Val or D-Val and X6represents Gln or D-Gln.

55. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 47, where X22the present is the focus of a Val or D-Val, or X 6represents Gln or D-Gln.

56. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 47, where only one of X10X11and X14represents Gly.

57. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 45 where the specified peptide is a

Lys-Leu-Lys-Lys-Gln-Leu-Ala-Asp-Leu-Leu-Arg-Glu-Leu-Leu-Gln-Glu-Phe-Leu-Glu-Leu-Val-Inp (SEQ ID NO:197); or

Lys-Leu-Lys-Lys-Gln-Leu-Ala-Asp-Leu-Leu-Arg-Glu-Leu-Leu-Gln-Glu-Phe-Leu-Glu-Leu-Val-Nip (SEQ ID NO:211), or

pharmaceutically acceptable salt of one of the above [connections].

58. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 47, where X10represents Gly, and X17represents Glu or D-Glu.

59. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 47, where each of X10X11and X14is other than Gly.

60. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 47, where X17represents Leu or D-Leu.

61. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 60, where X14represents Trp or D-Trp and X10represents Leu, Phe, D-Leu or D-Phe.

62. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 60, where X14the submitted is a Trp or D-Trp or X 10represents Leu, Phe, D-Leu or D-Phe.

63. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 45 where R1represents H and R2represents OH.

64. The peptide according to any of embodiments 1-63, where the peptide is presented in the form of pharmaceutically acceptable salts.

65. The peptide according to embodiment 64, where this salt is a metal salt or organic amine salt.

66. The peptide according to embodiment 65, where the specified metal is an alkaline metal or alkaline earth metal.

67. The peptide according to embodiment 65, where the specified metal is lithium, sodium, potassium, magnesium, calcium, aluminum or zinc.

68. The peptide according to embodiment 65, where the specified Amin represents triethylamine, ethanolamine, diethanolamine, triethanolamine, morpholine, N-methylpiperidine, N-ethylpiperidine or dibenzylamine.

69. The peptide according to embodiment 64, where this salt is a salt of accession acid.

70. The peptide according to embodiment 69, where the specified salt accession acid is a hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, sulfite, bisulfate, phosphate, acid phosphate, isonicotinate, lactate, salicylate, tartrate, bitartrate, ascorbate, getitemat, gluconate, glucuronate, saharat, format, benzoate, glutamate, Pantothenate, acetate, fumarate, with ccinet, methanesulfonate, aconsultant, bansilalpet, para-toluensulfonate, citrate or malate.

71. The peptide or pharmaceutically acceptable salt of the indicated peptide according to any of embodiments 1-63, where R1represents aminosidine group.

72. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 71, where the specified aminosidine group is dansyl; methoxycarbonyl; etoxycarbonyl; 9-fertilitycare; 2-chlorocarbons; 2,2,2-trichlorocyanuric; 2-fenilalaninammonii; tert-butoxycarbonyl; benzyloxycarbonyl; para-methoxybenzyloxy; para-nitrobenzenesulfonyl;o- nitrobenzisoxazole; para-bromobenzyloxycarbonyl; para-chlorobenzenesulfonyl; para-identilication; 2,4-dichlorobenzenesulfonyl; diphenylmethylene; 3,5 - dimethoxybenzonitrile; phenoxycarbonyl; 2,4,6-tri-tert-butylphenoxyacetyl; 2,4,6-trimethylbenzenesulfonyl; formyl; acetyl; chloroacetyl; trichloracetic; TRIFLUOROACETYL; phenylacetyl; pikolinos; benzoyl; para-phenylbenzyl; phthaloyl; methyl; tert-butyl; allyl; [2-(trimethylsilyl)ethoxy]methyl; 2,4-dimethoxybenzyl; 2,4-dinitrophenyl; benzyl; 4-methoxybenzyl; diphenylmethyl; triphenylmethyl; benzazolyl;o-nitrobenzenesulfenyl; 2,4-dinitrobenzenesulfonyl; para-toluensulfonyl; benzazolyl; 2,,6-trimethyl-4-methoxybenzenesulfonyl; 2,4,6-trimethoxybenzaldehyde; 2,6-dimethyl-4-methoxybenzenesulfonyl; pentamethylbenzene; 4-methoxybenzenesulfonyl; 2,4,6-trimethylbenzenesulfonyl; or benzylmethyl.

73. The peptide or pharmaceutically acceptable salt of the indicated peptide according to any of embodiments 1-63, where R2represents a carboxyl protective group.

74. The peptide or pharmaceutically acceptable salt of the indicated peptide according to embodiment 73, where the specified carboxyl protective group is methoxy; ethoxy; 9 fertilitate; methoxyethoxy; methylthiomethyl; tetrahydropyranyloxy; tetrahydrofuranate; methoxyethoxyethoxy; benzylacetone; phenacyloxy; pair-bromination; α-methylbenzylamine; pair-methoxybenzyloxy; depilarsi; 2 chloroethoxy; 2,2,2-trichloroethane, 2-methylthioethyl; 2-(para-toluensulfonyl)methoxy; tert-butoxy; cyclopentane; cyclohexane; allyloxy; metalliance; cinnamate; α-methylcinnamic; phenoxy; 2,6-dimethylphenoxy; 2,6-diisopropylphenol; benzyloxy; triphenylmethane; diphenylmethoxy; 2,4,6-trimethylaniline; pair-bromobenzylamine;o-nitrobenzyloxy; N,N-dimethylamide; pyrrolidinyl; or piperidinyl.

75. The peptide or pharmaceutically acceptable salt of the indicated peptide according to any of embodiments 1-63, where one or more of-NH2- or COOH-groups of the indicated peptide reveal who I am protected by a protective group.

76. A composition comprising an effective amount of the peptide or pharmaceutically acceptable salt of the indicated peptide according to any of embodiments 1-75, and a pharmaceutically acceptable carrier or excipient.

77. A method of treating or preventing dyslipidemia, involving the administration to a mammal, in case of need, an effective amount of the peptide or pharmaceutically acceptable salt of the indicated peptide according to any of embodiments 1-75.

78. Method according to embodiment 77, where this dyslipidemia is hyperproteinemia high concentration of serum low-density lipoprotein, and high concentration in serum lipoprotein very low density, hyperlipidemia, low concentrations of high density lipoprotein in serum, hypocholesterolemia, A-beta lipoproteinemia, deficiency of ApoA-I and Tangier disease.

79. Method according to embodiment 77, where this dyslipidemia is a hyperlipidemia, hypercholesterolemia, deficiency of ApoA-I or hypertriglyceridemia.

80. Method according to embodiment 77, where said treatment involves increasing the concentration of high density lipoprotein in the serum.

81. A method of treating or preventing cardiovascular disease, involving the administration to a mammal, if necessary, effectively the positive amount of the peptide or pharmaceutically acceptable salt of the indicated peptide, according to any of embodiments 1-75.

82. Method according to embodiment 81, where cardiovascular disease is a metabolic syndrome, coronary heart disease, atherosclerosis, restenosis, groove toxins, congestive heart failure, circulatory shock, cardiomyopathy, heart transplantation, myocardial infarction, cardiac arrhythmia, supraventricular tachycardia, atrial flutter, paroxysmal atrial tachycardia, aneurysm, angina, acute violation of cerebral circulation, peripheral vascular disease, cerebrovascular disease, renal disease, atherogenesis, atherosclerosis, acute pancreatitis, and coronary arteries.

83. Method according to embodiment 81, where cardiovascular disease is a atherosclerosis, restenosis or metabolic syndrome.

84. A method of treating or preventing endothelial dysfunction, involving the administration to a mammal, in case of need, an effective amount of the peptide or pharmaceutically acceptable salt of the indicated peptide according to any of embodiments 1-75.

85. A method of treating or preventing macrovascular disorder, involving the administration to a mammal, in case of need, an effective amount of the peptide or pharmaceutically acceptable salt of the indicated peptide, according to the SNO any of embodiments 1-75.

86. Method according to embodiment 85, where the specified macrovascular disorder is a transient ischemic stroke, stroke, angina, myocardial infarction, heart failure or peripheral vascular disease.

87. A method of treating or preventing microvascular disorders, involving the administration to a mammal, in case of need, an effective amount of the peptide or pharmaceutically acceptable salt of the indicated peptide according to any of embodiments 1-75.

88. Method according to embodiment 87, where the specified microvascular disorder is a diabetic retinopathy, microalbuminuria, macroalbuminuria, end-stage renal disease, erectile dysfunction, autonomic neuropathy, peripheral neuropathy, osteomyelitis or ischemia of the lower limb.

89. Method according to any of embodiments 77-88, according to which the mammal is a human.

90. Method according to any of embodiments 77-89, according to which the introduction is carried out orally, intravenous, intramuscular, intrathecal, subcutaneous, sublingual, nasal, cutaneous, transdermal, ocular route or by inhalation.

1. The peptide, which are mimetic ApoA-I having the following amino acid sequence:
Lys-Leu-Lys-Gln-Lys-Leu-Ala-Glu-Leu-Leu-GluAsn-Leu-Leu-Glu-Arg-Phe-Leu-Asp-Leu-Val-Inp (SEQ ID NO: 16) ,
or its pharmaceutically acceptable salt.

2. The peptide/lipid complex intended for the treatment of dyslipidemia, cardiovascular disease, endothelial dysfunction, macrovascular disorders and microvascular disorders, containing a phospholipid and peptide or pharmaceutically acceptable salt of the peptide according to claim 1, where the phospholipid is one or more of sphingomyelin, DPPC and DPPG and the ratio of the total number of peptide to phospholipid is from about 1 : about 0.5 to about 1 : about 5.

3. The peptide/lipid complex of claim 2, where the phospholipid is a mixture of sphingomyelin and DPPC or DPPG.

4. The peptide/lipid complex of claim 2, where the phospholipid is a mixture of sphingomyelin, DPPC and DPPG.

5. The peptide/lipid complex according to claim 4, where the mass ratio of peptide : sphingomyelin : DPPC : DPPG is 1 : 1,2125 : 1,2125 : 0,075.

6. The peptide/lipid complex according to claim 4, where the mass ratio of sphingomyelin : DPPC : DPPG is 48,5 : 48,5 : 3.

7. The peptide/lipid complex of claim 2, where the mass ratio of the total number of peptide to phospholipid is 1 : 2,5.

8. The peptide/lipid complex of claim 2, where the phospholipid is a mixture of sphingomyelin, DPPC and DPPG, the mass ratio of peptide : sphingomyelin : DPPC : DPPG is 1 : 1,2125 : 1,2125 : 0,075, and the mass ratio of peptide to lipid sostavljaet : 2,5.

9. The composition is intended for treating or preventing dyslipidemia, cardiovascular disease, endothelial dysfunction, macrovascular disorder or microvascular disorders, containing the peptide/lipid complex according to any one of paragraphs. 2-8 in the amount effective for treating or preventing dyslipidemia, cardiovascular disease, endothelial dysfunction, macrovascular disorder or microvascular disorder, and a pharmaceutically acceptable carrier or excipient.

10. A method of treating or preventing dyslipidemia, providing an introduction to the needy in the mammal a peptide/lipid complex according to any one of claim 2 to 8 in number, effective for the treatment of dyslipidemia.

11. A method of treating or preventing cardiovascular disease, providing an introduction to the needy in the mammal a peptide/lipid complex according to any one of claim 2 to 8 in number, effective for the treatment and prevention of cardiovascular diseases.

12. A method of treating or preventing endothelial dysfunction, providing an introduction to the needy in the mammal a peptide/lipid complex according to any one of claim 2 to 8 in number, are effective for treatment and prevention of endothelial dysfunction.

13. A method of treating or pre which pre-emption macrovascular disorder, introducing the needy in the mammal a peptide/lipid complex according to any one of claim 2 to 8 in an amount effective for treating or preventing macrovascular disorders.

14. A method of treating or preventing macrososudistah disorders, providing an introduction to the needy in the mammal a peptide/lipid complex according to any one of claim 2 to 8 in an amount effective for treating or preventing microvascular disorders.



 

Same patents:

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to immunology and medicine, namely to new peptides of general formula: X-CC-C(Y)-CC-Z, wherein X is a B-cell epitope of apolipoprotein B100 protein; C is an amino acid residue specified in K or R; Y is an immunoadjuvant specified in a group of Pam1CSS-; Pam2CSS-, Pam3CSS-; Z - T-helper epitope specified in AKFVAAWTLKAAA, KLIPNASLIENCTKAEL, QYIKANSKFIGITE.

EFFECT: peptide preparations possess higher antisclerotic activity as compared to their analogues, and the compositions thereof are of practical interest as a vaccine for preventing and treating atherosclerotic vascular disease.

9 cl, 1 dwg, 6 tbl, 7 ex

FIELD: chemistry.

SUBSTANCE: method of purifying apoliprotein A-1 involves mixing fractions of plasma IV, obtained via low temperature-ethanol fractionation with 1-8 M urea solution to form a prepared solution of fraction IV; feeding the prepared solution into a first column for anionic chromatography and subsequent elution with 1-8 M urea solution to obtain a solution of aroA-1 protein; and feeding the solution of aroA-1 protein into a second column for anionic chromatography and elution with 0-1 M urea solution to obtain pure aroA-1 protein.

EFFECT: higher purity.

26 cl, 7 dwg, 6 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to novel peptides, which possess ability to relieve at least one symptom of atherosclerosis.

EFFECT: peptides have high stability, it is easy to introduce them in per oral way.

8 cl, 31 dwg, 17 tbl, 5 ex

The invention relates to medicine and can be used to obtain the dimer molecular variants of APO-lipoprotein AI-Milano and use it for the manufacture of medicaments for the prevention and treatment of atherosclerosis and cardiovascular disease

FIELD: medicine.

SUBSTANCE: on the first experimental day, cardiopathy is simulated by a single subcutaneous administration of equally portioned mixture of native egg albumin and Freund's complete adjuvant in rats. The mixture is administered at 0.2 ml of the mixture into 5 injection points: abdominally, into inguinal and axillary regions on the left and right. The cardiopathy is prevented by daily gastric administration of succinic acid 1.5 mmole/kg for 60 days through a probe.

EFFECT: higher clinical effectiveness.

2 dwg, 1 tbl, 1 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to the pharmaceutical industry and medicine. What is presented is using treo-3-phenylglutamic acid hydrochloride of the following structural formula: as an agent possessing the cardioprotective, antiplatelet, anticoagulant and membrane-protective properties under stress stimulation.

EFFECT: compound possesses the high cardioprotective, antiplatelet, anticoagulant and membrane-protective activity.

9 tbl

FIELD: chemistry.

SUBSTANCE: invention relates to novel compound of salvianolic acid L with general formula (I) , to its pharmaceutically acceptable salts and hydrolysable ethers, with compound of salvianolic acid L having one pair of protons of trans-form double bond and one proton of single-substituted double bond; and compound of salvianolic acid L is intended for treating cardiovascular disease, capture of free radicals and/or prevention of excessive oxidation.

EFFECT: invention also relates to method of its preparation, medication, which contains salvianolic acid L, and its application for preparation of drug for treatment of cardio-cerebral-vascular diseases.

13 cl, 13 dwg, 17 tbl, 10 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to compound of formula (I) , where ring A is cycloalkane ring with number of members from 3 to 7, benzene ring or monocyclic 5-member or 6-member aromatic heterocyclic ring, containing 1 heteromember of ring, selected from the group, containing N and S, and benzene and heterocyclic rings can optionally have one or two similar or different substituents, selected from the group, containing halogen, HO-, R1-O-, H2N-C(O)- and NC-; Y is selected from the group, containing S, C(R12)=C(R13) and C(R15)=N; Z is selected from the group, containing C(R16); R1, R30, R33, R35, R54 and R55 independently on each other group R1, R30, R33, R35, R54 and R55 are selected from the group, containing (C1-C6)-alkyl, (C2-C6)-alkenyl, (C3-C7)-cycloalkyl and (C3-C7)-cycloalkyl-(C1-C4)-alkyl-, and all of them can optionally have one or more similar or different substituents R70; R3 and R5 represent hydrogen; R4 and R6 are selected independently on each other, from the group, containing hydrogen and , (C1-C4)-alkyl; R12, R13, R15 and R16 are selected independently on each other, from the group, containing hydrogen, halogen and O2N-; R20 is selected from the group, containing hydrogen and (C1-C4)-alkyl; one of the groups R21 and R22 is group of formula II: R24-R23-, and the other of groups R21 and R22 is selected from the group, containing hydrogen, halogen, R30, HO-, R30-O-, R30-S(O)m-, H2N-, R30-NH-, R30-N(R30)-, R30-C(O)- and NC-; R23 is chain, containing from 1 to 5 chain members of which 0 or 1 chain member is heteromember of chain, selected from the group, containing N(R25), O, S, with other chain members being similar or different groups C(R26)(R26), where two adjacent groups can be bound to each other by double bond; R24 is selected from the group, containing hydrogen, R31, R31-O-, R31-NH-, R31-N(R31)-, R31-C(O)-NH-, HO-C(O)- and monocyclic, bicyclic or tricyclic ring with number of members from 5 to 10, which is saturated or non-saturated and contains 0, 1, 2 or 3 similar or different ring heteromembers, selected from the group, containing N, N(R32), O, S, and ring can optionally have on ring carbon atoms one or 2-3 similar or different substituents, selected from the group, containing halogen, R33, R33-O-, R33-S(O)m-, R33-C(O)-NH-, R33-S(O)2-NH-, R33-C(O)-, HO-C(O)-, H2N-C(O)-, R33-NH-C(O)-, R33-N(R33)-S(O)2-, NC-, oxo, phenyl and Het; on condition that total number of C, N, O and S atoms, present in two groups R23 and R24, constitutes not less than 5; R25 is selected from the group, containing hydrogen and (C1-C4)-alkyl; R26, independently on each other group R26, is selected from the group, containing hydrogen, fluorine, (C1-C4)-alkyl and HO-, or two groups R26, together with included into them chain members, form monocyclic ring with number of members 4, which is saturated and contains 1 ring heteromember, selected from the group, containing O; R31 is selected from the group, containing (C1-C6)-alkyl, which can optionally have one substituent R70; R32 is selected independently on each other, from the group, containing hydrogen, R35 and phenyl; R50 is selected from the group, containing R51-O- and R52-N(R53)-; R51 is selected from the group, containing hydrogen and R54; R52 is selected from the group, containing hydrogen; R53 is selected from the group, containing hydrogen; R70 is selected from the group, containing HO-, R71-O-, H2N-, R71-NH-, R71-N(R71)-, R71-C(O)-NH-, HO-C(O)-, H2N-C(O)- and phenyl; R71, independently on each other group R71, is selected from the group, containing (C1-C4)-alkyl; Het, independently on each other group Het, is monocyclic heterocyclic ring with number of members 5, which contains 1 or 2 similar or different ring heteromembers, selected from the group, containing N and S, and ring is saturated or non-saturated and optionally substituted with one or more similar or different substituents, selected from the group, containing (C1-C4)-alkyl; m, independently on each other number m, is integer number, selected from the group, containing 0, 1 and 2; phenyl, independently on each other phenyl group, can optionally have one or more similar or different substituents, selected from the group, containing halogen and (C1-C4)-alkyl.

EFFECT: invention also relates to method of obtaining compound of formula (I) and its application for manufacturing pharmaceutical for inhibiting receptor Edg-2.

17 cl, 14 tbl, 362 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutics, namely to a formulation for oral transmucosal administration which contains a lipid-lowering active substance specified in statins, fibrates or ezetimibe; an aqueous-alcohol solution consisting of water and 30° to 70° ethanol, wherein the above active substance is found in a stable and completely dissolved state with the pH value of the formulation falling within the range of 5.0 to 8.0. The invention also refers to a method for preparing the above formulation and using it for treating hyperlipidemia.

EFFECT: invention provides better efficacy for lower doses of the active substances (statins, fibrates and ezetimibe); their immediate bioavailability for hepatocytes and considerably reduced production of harmful metabolites that stands for eliminating the main side effects of these agents.

11 cl, 6 ex

Pcsk9 antagonists // 2528735

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to immunology and biotechnology. There are presented: an isolated antibody or its variant specifically recognising PCSK9, and a based pharmaceutical composition for lowering LDL-cholesterol. There are described: using for lowering blood cholesterol and/or LDL; reducing an incidence rate and/or correcting abnormal cholesterol and/or LDL; using for preparing a drug for lowering blood cholesterol and/or LDL; reducing the incidence rate and/or correcting abnormal cholesterol and/or LDL. There are disclosed versions of cell lines producing PCSK9 antibody or its antigen-binding portion and deposited in the American typical culture collection (ATCC) under Nos. PTA-8986, ATCC PTA-8984, ATCC PTA-8983, respectively. What is described is a coding nucleic acid and a host cell for producing the based antibody.

EFFECT: invention provides PCSK9 agonist antibodies that can find application in medicine for lowering cholesterol.

18 cl, 24 dwg, 9 tbl, 9 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to compounds of formula (I), wherein A means morpholinyl, 1,4-oxazepamyl, piperidinyl, pyrrolidinyl or azetidinyl which is bound to N; R1 means C1-C6-alkyl group; R2 means bicyclic aryl group specified in 1H-indolyl, 1H-pyrrolo[3,2-b]pyridyl, quinolyl, naphthyl, 1H-pyrrolo[2,3-b]pyridyl, 5H-pyrrolo[3,2-d]pyrimidinyl, 7H-pyrrolo[2,3-d]pyrimidinyl, benzo[b]thiophenyl, imidazo[1,2-a]pyridyl, benzo[b]thiazolyl, 5H-pyrrolol[2,3-b]pyrazinyl and quinoxalinyl which can be substituted by R4; R3 means hydrogen or halogen atom; R4 means C1-C6-alkyl group, C1-C6-halogenalkyl group, OR1A, halogen, -(CH2)aOH, CN, NHCOR1A, SO2R1A or NHSO2R1A; R5 means C1-C6-alkyl group, -(CH2)aOH, -(CH2)aOR1B, halogen or CONH2; provided p is a plural number, R5 can be identical or different, or R5 can be combined with another R5; each of R1A and R1B independently means C1-C6-alkyl group; a is equal to 0, 1 or 2; n is equal to 1 or 2; p is equal to 0, 1, 2, 3, 4 or 5. Besides, the invention refers to intermediate compounds of formulas (IA) and (IB) for preparing the compounds of formula (I), to a preventive or therapeutic agent containing the compounds of formula (I), pharmaceutical compositions, using the compounds of formula (I) and to a method for preventing or treating diseases.

EFFECT: compounds of formula (I) as selective 5-HT2B receptor antagonists.

11 cl, 1 dwg, 18 tbl, 88 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a new compound of formula [I] or to its pharmaceutically acceptable salt, wherein A represents optionally substituted alkyl, wherein the substitute represents identical or different 1-3 groups specified in aryl optionally substituted by 1-3 groups specified in alkyl, halogen, alkoxy and alkanoyl; cycloalkyl optionally substituted by 1-3 groups specified in alkyl and halogen; hydroxy; alkoxy; halogen; an amino group and oxo; an optionally substituted carbocyclic group specified in a mono- and bicyclic group, wherein an aromatic ring and cycloalkyl are condensed; optionally substituted aryl, an optionally substituted completely saturated 5- or 6-merous monocyclic heterocyclic group each of which contains 1 heteroatom specified in nitrogen and oxygen, wherein the substitute of optionally substituted aryl, the optionally substituted carbocyclic group and the optionally substituted heterocyclic group for A represents identical or different 1-3 groups specified in alkyl, optionally substituted hydroxy, alkoxy, cycloalkyl or halogen; cycloalkyl optionally substituted by alkyl or alkoxy; alkoxy optionally substituted by halogen; halogen; hydroxy; oxo; heterocycle; alkyl sulphonyl; and mono- or dialkylcarbamoyl, optionally substituted amino, wherein the substitute represents identical or different 1 or 2 alkyl or aryl, or optionally substituted carbamoyl, wherein the substitute represents identical or different 1 or 2 alkyls optionally substituted by aryl, X represents optionally substituted methylene or -O-, wherein the substitute of optionally substituted methylene for X represents alkoxy or hydroxy, Q represents N or C-R4, L1 represents a single bond, methylene, -CH=CH-, -O-, -CO-, -NR11-, -NR11CO-, -CONR11- or -CH2NR11-, L2 represents a single bond, -CR6R7- or a bivalent 5- or 6-merous completely saturated monocyclic heterocyclic group each of which contains 1 heteroatom specified in nitrogen and oxygen, R1 and R2 are identical or different, and each represents hydrogen, alkyl or halogen, R3 and R4 are identical or different, and each represents hydrogen, alkyl, alkoxy, cyano or halogen, R1 and R3 are optionally bond thereby forming 5- or 6-merous cycloalkane, or a 5- or 6-merous aliphatic heterocycle containing oxygen atom, R5 represents a carboxyl group, an alkoxycarbonyl group or a bioisosteric group of the carboxyl group, R6 and R7 are identical or different, and each represents hydrogen or alkyl, or R6 and R7 are bond thereby forming cycloalkane, R8 represents hydroxy, alkanoylamino or alkyl sulphonylamino, R9 and R10 represent hydrogen or halogen, and R11 represents hydrogen or alkyl. Besides, the invention refers to specific compounds of formula [I], a drug based on the compound of formula [I], using the compound of formula [I], a method of treating based on using the compound of formula [I], and an intermediate compound of formula [II].

EFFECT: there are prepared new compounds possessing the agonist activity on thyroid hormone β receptor.

18 cl, 36 tbl, 344 ex

FIELD: chemistry.

SUBSTANCE: invention relates to compounds, capable of providing uncoupling of respiration of mitochondria in direct dependence on the value of their membrane potential and causing mild uncoupling - reversible reduction of the membrane potential of the mitochondria, which does not cause substantial/physiologically meaningful suppression of respiration, an injury to the respiratory chain components or impairment of physic-chemical properties of mitochondrial membranes, and can be applied in medicine and biology. The said compound has a structure , where R1 and R3 are hydrogen atoms, R2 and R4 are hydrogen atoms independently on each other or either substituted or non-substituted C1-C4 alkyl groups; R2, bound with R5; or R4 with R6 form either substituted or non-substituted 1,3-propylene; R5, R6, R7 and R8 independently on each other represent a hydrogen atom or methyl; n=1; A is pharmacologically acceptable anion.

EFFECT: claimed are the mild uncouplers and based on them compositions for application in biology and medicine.

15 cl, 7 dwg, 6 ex

FIELD: medicine.

SUBSTANCE: waist circumference (WC), height and weight are measured to calculate a body weight index (BWI), to determine glucose and lipids. The WC more than 80 cm requires prescribing a moderately hypocaloric diet that implies limiting an energy value of food ration by 500-600 kkal a day with the daily food ration composed taking into account as follows: total fat consumption less than 30% of total energy consumption; carbohydrates - up to 50%; protein content in the food ration - 15-20%; dietary fibre - 20-30 mg; fresh fruit and vegetables - 400-500 g; nuts, cereals, beans - 30 g; table salt - less than 5 g; 1000 mg of calcium a day for females taking hormonal contraception or replacement hormonal therapy (RHT), and 1500-2000 mg in females taking no hormonal contraception or RHT, vitamin D - 800 IU, polysaturated fatty acids 0.5-1.0 g, folic acid - 400 mcg, vitamin C - 60 mcg, vitamin E - 30 Units; physical exercises - not less than 40 min a day and not less than 5 times a week; alternating aerobic and anaerobic exercises. If BWI falls within the range of 25 to 29.9 kg/m2, a normal glucose and lipid level, combined oral contraceptives (COC) with natural oeastrogen and dienogest, or the COC with drosperinone are taken. The BWI within the range of 30 kg/m2 and more, the normal glucose and lipid level, pure progestin oral contraceptives (PPOC) are administered. If the BWI exceeds 25 kg/m2, and fasting hyperglycemia, impaired glucose tolerance (IGT) and/or dyslipidemia are observed, pure progestin oral contraceptives (PPOC) are administered. If a postmenopausal female has the BWI less than 30 kg/m2 and suffers climacteric syndrome, a combined replacement hormonal therapy (RHT) containing drospirenone is prescribed. The BWI more than 30 kg/m2 requires administering meldonium into postmenopausal females.

EFFECT: method enables individual prevention of oestrogen-dependent diseases.

6 cl, 14 tbl, 2 ex

FIELD: medicine.

SUBSTANCE: haemodilution is performed with using crystalloid solutions, nitroglycerin and antihypertensive drugs. The antihypertensive drug is presented by using 5% pentamine intermittently administered after the beginning of the anaesthetic induction; if the effect of pentamine occurred to be inadequate, nitroglycerin is infused in an amount of 10 ml as a 0.1% solution diluted in crystalloid solution 200 ml for maintaining reference arterial blood pressure of 80/50 mm Hg. The nitroglycerin solution is kept to be administered until the wound is sutured, while the additional antihypertensive and blood loss reducing effect is ensured by spinal anaesthesia; as a haemostatic agent, an officinal solution of tranexamic acid 5-10 ml is intraoperatively intravenously introduced under an endoprosthesis bed.

EFFECT: lower risk of postoperative complications and ensured controlled stable arterial blood pressure in the patient.

2 ex

FIELD: medicine.

SUBSTANCE: invention refers to biotechnology, more specifically to modified von Willebrand factor (VWF), and can be used in medicine. A recombinant method is used to preparing modified VWF fused in C-terminal of its primary translation product with N-terminal of albumin by the linker SSGGSGGSGGSGGSGGSGGSGGSGGSGGSGS. The prepared modified VWF is used as a part of the pharmaceutical composition for treating or preventing coagulation failure.

EFFECT: invention enables preparing the modified VWF which maintains its ability to N-terminal dimerisation and C-terminal multimerisation with a prolonged half-period of functional blood plasma occurrence as compared to the half-period of functional VWF occurrence.

17 cl, 5 dwg, 4 tbl, 11 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to 6-substituted isoquinoline and isoquinolinone derivatives of formula or to its stereoisomer and/or tautomer forms and/or a pharmaceutically acceptable salt, wherein R1 represents H, OH or NH2; R3 represents H; R4 represents H, a halogen atom, CN or (C1-C6)alkylene-(C6-C10)aryl; R5 represents H, a halogen atom, (C1-C6)alkyl; R7 represents H, a halogen atom, (C1-C6)alkyl, O-(C1-C6)alkyl; R8 represents H; R9 and R6 are absent; R10 represents (C1-C6)alkyl, (C1-C8)heteroalkyl, (C3-C8)cycloalkyl, (C6)hetrocycloalkyl, (C1-C6)alkylene-(C3-C8)cycloalkyl, (C1-C6)alkylene-(C6-C10)aryl, (C1-C6)alkylene-(C6)heterocycloalkyl; R11 represents H; R12 represents (C1-C6)alkyl, (C3-C8)cycloalkyl, (C5)heteroaryl or (C6-C10)aryl; R13 and R14 independently represent H, (C1-C6)alkyl, (C1-C6)alkylene-R'; n is equal to 0; m is equal to 2 or 3; s is equal to 1 or 2; r is equal to 1; L represents O or NH; R' represents (C3-C8)cycloalkyl, (C6-C10)aryl; wherein in the rests, R10, R12-R14 alkyl or alkylene are unsubstituted or optionally substituted by one or more OCH3; wherein in the rests, R10, R12-R14 alkyl or alkylene are unsubstituted or optionally substituted by one or more halogen atoms; wherein (C1-C8)heteroaryl group means (C1-C8)alkyl groups, wherein at least one carbon atom is substituted by O;. (C6)heterocycloalkyl group means a monocyclic carbon ring system containing 6 ring atoms wherein one carbon atom can be substituted by 1 oxygen atom or 1 sulphur atom which can be optionally oxidated; (C5)heteroaryl means a monoring system wherein one or more carbon atoms can be substituted by 1 nitrogen atom or 1 sulphur atom or a combination of various heteroatoms. Also, the invention refers to using the compound of formula (I) and to a therapeutic agent based on the compound of formula (I).

EFFECT: there are prepared new compounds effective for treating and/or preventing diseases associated with Rho-kinase and/or mediated by Rho-kinase by phosphorylation of myosin light chain phosphatase, and the compositions containing these compounds.

32 cl, 111 ex

FIELD: medicine.

SUBSTANCE: to correct haemostatic disorders in case of chronic calculous cholecystitis at the background of chronic hepatitis or cirrhosis of liver, preoperative correction of haemostasis with the preparation prothromplex 600, introduced intravenously at a rate not higher than 2 ml/min in a dose of 20 IU/kg of the patient's body weight, is carried out 3 days before endosurgical treatment. After that, on the first day after operation 20 IU/kg of prothromplex 600 and additionally 10 mg/kg of etamsylate and 25 mg/kg of aminomethyl are introduced intravenously. The preparations are introduced 1 time per day for 3 days with obligatory control of haemostasis indices.

EFFECT: method makes it possible to enhance the clinical effect due to an improvement of haemostasis system indices, connected with an increase of quantitative indices of a coagulation link, at the background of normalisation of the aggregation activity of platelets, as well as an arrest of hypocoagulation due to normalisation of the prothrombin complex, which makes it possible to twice reduce the treatment duration.

2 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: method involves intravenous allogeneic transplantation of multipotent mesenchymal stromal cells recovered from the placenta in an amount of 6 mln cells/kg. Additionally, haemopoietic stem cells recovered from umbilical blood in an amount of 300 thousand cells/kg are introduced.

EFFECT: method enables reducing the number of cytogenetically changed cells, promotes activating myeloid tissue regeneration in old laboratory animals.

4 tbl, 1 ex

Up!