Treatment and prevention method of gram-positive bacteria induced diseases

FIELD: medicine.

SUBSTANCE: invention belongs to medicine, notably to pharmaceutical composition for treatment and prevention of bacterial infections induced by gram-positive bacteria. Composition includes effective doses of cholanic acid or its salt, phosphatidylcholine and neutral lipids. Neutral lipids and phospholipids are associated in lipoprotein-like particles, which does not include proteins or peptides.

EFFECT: composition effectivity is caused by its ability to bind lipoteichoic acid of gram-positive bacteria, neutralising or preventing their pathogenic action.

7 cl, 2 tbl, 1 ex

 

The technical FIELD

The present invention relates to the treatment of infections caused by Gram-positive bacteria. In particular, the present invention relates to the treatment of infections by the introduction of various compositions, which have the ability to neutralize and/or remove toxins of Gram-positive bacteria from the body, in addition, the present invention relates to a method of prophylaxis using these compositions. Preferably, these compositions contain bile acids or bile salts, such as Galanova acid or salt holonovel acid, a neutral lipid, such as a triglyceride or a phospholipid, such as phosphatidylcholine, and used for the treatment or prevention of pathological conditions due to Gram-positive bacteria, such as (without limitation specified) sepsis, septic shock syndrome, systemic inflammatory response (SIRS), SIRS with organ dysfunction and/or failure, organ dysfunction and organ failure.

The LEVEL of TECHNOLOGY

Normal serum contains a number of lipoprotein particles, which are classified on the basis of density and divided into chylomicrons, lipoproteins, very low density lipoproteins (VLDL), LDL, low Vlastnosti (LDL) and high density lipoprotein (HDL). They consist of a Saint is free and esterified cholesterol, triglycerides, phospholipids and other lipid components and protein. Lonp carry energy in the form of triglycerides to cells of the body where they are stored and used. After delivery of triglycerides lonp transformed into LDL. LDL delivers cholesterol and other lipid soluble materials to the cells of the body, while HDL carry excess or unused lipids to the liver for disposal. In the norm all of these lipoproteins balanced, ensure timely delivery and removal of lipid soluble materials. Abnormally low levels of HDL cholesterol can lead to the development of a wide range of pathological conditions, as well as cause secondary complications of various diseases.

Under normal conditions HDL are solid particles, the surface of which is covered with a phospholipid monolayer that surrounds the hydrophobic core. Apolipoprotein A-I and A-II is attached to the surface by hydrophobic interaction plots their alpha-helical domains. In just the secreted form of the particle is a disk containing three balance of cholesterol in the bilayer. Cholesterol tarifitsiruetsya using enzyme lecithincholesterolacyltransferase (LAT) and transferred to the center of the disk. The movement of ester cholesterol in the center of the disc is the result space is n limits the solubility of the bilayer. The HDL particle as esterification and movement of cholesterol in the centre adopts a spherical shape. Ester of cholesterol and other water-soluble lipids, which are collected in "nabusasa the core of HDL cholesterol are then excreted by the liver.

Jonas et al., Meth. Enzym. 128A: 553-585 (1986) managed to get a number of reconstructed particles, reminiscent of HDL. The method includes the selection of HDL cholesterol and lipid removal of them by using standard methods (Hatch et al., Adv. The Lip. Res. 6: 1-68 (1968); Scanu et al., Anal. Biochem. 44:576-588 (1971) to get APO-HDL particles. Apalaci fractionary and restore the phospholipid (with or without cholesterol) using detergent dialysis.

Matz et al., J. Biol. Chem. 257 (8):4535-4540 (1982) describe the micelle phosphatidylcholine with apolipoprotein A1. Describe the various relationships of these components, it is assumed that the described method can be used for other micelles. It also assumes that it is possible to use molecules as enzyme substrate or to simulate the HDL molecules. However, in this application does not discuss the possibility of using these micelles to remove cholesterol, as well as in diagnostic or therapeutic purposes.

Wiliams et al., Biochem &Biophys. Acta 875:183-194 (1986) describe phospholipid liposomes, which when injected into the plasma capture apalaci and cholesterol. The authors describe liposomes, who ohvatyvajut apalaci and cholesterol in vivo, and assume that the capture of cholesterol increases in the case of phospholipid liposomes pre-captured apalaci.

Williams et al., Persp. Biol. & Med. 27(3):417-431 (1984) describe the possibility of using lecithin liposomes to remove cholesterol. In this paper summarizes earlier data showing that liposomes that contain apalaci remove cholesterol from cells in vitro more effectively than liposomes that do not contain apalaci. However, in this paper does not discuss the possibility of using liposomes or micelles in vivo and suggests that in any job in vivo with liposomes needed extreme caution.

In U.S. patents№5,506,218; 5,344,822; 5,614,507; 5,587,366; 5,674,855 described as compositions containing phospholipids, such as phosphatidylcholine, natural lipid, such as triglyceride and bile acids and bile salts, such as Galanova acid and its salt, for example Holt sodium, can be used for the prevention and treatment of infections caused by Gram-negative bacteria, such as S. typhimurium, due to the inactivation of anchor molecules of the lipid a of the lipopolysaccharide (LPS).

In U.S. patent No. 5,128,318 describes that the recovered particles containing HDL-associate apalloc and lipid, are able to bind endotoxin and inactivate it can be used as effective the second material to reduce toxicity, caused by endotoxin.

Thus, it was found that phospholipids can be used alone or in combination with additional materials, such as neutral lipids, bile salts, etc. as effective agents for the prevention and/or suppression of infections caused by Gram-positive bacteria. Especially preferable to use phosphatidylcholine (hereinafter, PC), both independently and in combination with other phospholipids, such as sphingomyelinase, in compositions that contain virtually no protein and peptides, such as apalaci or peptides based on them. Neutral lipids, such as mono-, di - and triglycerides, can be used in combination with phospholipids, provided that the total content of neutral lipids below a certain weight%, when the composition is used for intravenous bolus injection. In the case of other routes of administration, such as intravenous drip, the weight percentage is not so critical, but nevertheless it is desirable to take into account. Salts of bile acids or bile salts, such as holty, for example Holt sodium, can be used in combination with one or more components to obtain the most effective compositions.

The most preferred domestic the present invention are compositions, in which the neutral lipid is a triglyceride, ester cholesterol, or a combination of ester cholesterol and triglycerides.

This description reveals the effectiveness of bile acids, bile salts, such as holty, and/or neutral lipids, such as phosphatidylcholine and/or triglycerides for the treatment, prevention and/or prevent the development of infections caused by Gram-positive bacteria. These bile acids can be used alone or in combination with one or more phospholipids and/or neutral lipids, such as phosphatidylcholine and/or triglycerides. The claimed compositions can be used for treatment or prevention of pathological conditions (no limit specified), described above, preferably, use of the composition described above, in the form of an emulsion.

A DETAILED DESCRIPTION of the PRESENT INVENTION

EXAMPLE

In the experiments described below, it was shown that the emulsion containing phosphatidylcholine, triglycerides and Holt sodium is effective for removing toxins of Gram-positive bacteria from the blood.

Get emulsion containing solution of phosphatidylcholine (PC) at a concentration of 99.7 mg/ml, 18 mm cholate sodium and triglycerides (TG), which account for 7.5% by weight of the total weight of the lipid emulsion, an aqueous solution of the glycerol (2.6%).

As a control, use a 2.6% solution of glycerol.

The emulsion and the control was diluted to a concentration of 1:10 and added to esterwegen solutions blood treated with EDTA at 50% dilution; the solution is gradually added a solution (5 mg/ml of various dilutions) lipoteichoic acid (LTA). Lipoteicholic Acid)isolated from B. subtilis.

The samples are mixed and incubated at 37°C for 4 hours, then cooled on ice. The samples centrifuged (1000 RPMs, 2000 xg), and then measure the concentration of tumor necrosis factor (TNF) using standard commercially available ELISA.

The results presented in Table 1 (emulsion) and Table 2 (control), showing the efficiency of emulsions for removal of the toxin from the blood.

Table 1
Emulsion
LTATNF
ng/mlPG/ml
0.0010
0.0100
0.1000
0.3000
1.000 4.531
3.0000
Of 10,0000
100.00015.15

Table 2
Control, glycerol
LTATNF
ng/mlPG/ml
0.0010
0.0100
0.10044.19
0.3008.081
1.000At 66.41
3.00058.07
Of 10,00039.4
100.000290.7

The examples below give a detailed description of the invention, which, according to one aspect, relates to a method for preventing sepsis, septic shock syndrome, systemic inflammatory response (SIRS), with SIRS dysfunction/nedostatocnost the Yu bodies, dysfunction and organ failure caused by Gram-positive bacteria.

In some examples show that the introduction of family members bile acids or bile salts, for example holonovel acid or a salt thereof, in combination with a phospholipid or a neutral lipid, or only with the phospholipid may be used, for example, for the prevention, suppression, prevention or treatment of infections caused by Gram-positive bacteria. Thus, compositions not containing peptides and proteins, including only the bile acid/salt of the bile acid or the specified connection in combination with a phospholipid, can be used to treat these infections. Holomovie acids are described, for example, Hofmann, Hepatology 4(5):4S-14S (1984). Special attention is drawn to page 5S, Figures 1 and 2, which presents structural characteristics holanovich acids.

Subject to this treatment preferably is, however, an object of the present invention can be used in veterinary practice.

The term "inhibition"as used here, refers to treatment that focuses on reducing the severity of infection caused by any of a variety of toxins produced by Gram-positive bacteria (e.g., B. subtilis). Prevention can be done by introducing the agent into the mo is UNT, when the subject is in a situation of possible exposure to Gram-positive bacteria. This situation typically occurs during surgical interventions. Thus, the subject, which is surgical treatment, the active ingredient may be administered before the operation.

An effective amount of a combination of phospholipid and bile acids needed for treatment can vary. In General, the total dose is preferably from 200 mg to 800 mg of phospholipid per kg of body weight of the subject, the dose may decrease or increase depending on the severity of infection and the degree of risk, if the agent is introduced to prevent. For bile acids and their salts, such as holaaaaa acid and its salt, an effective dose varies approximately from 10 mg to 300 mg/kg of body weight of the subject, more preferably approximately from 15 to 275 mg / kg of body weight of the subject.

It is advisable to enter the bile acids/salts of bile acids and phospholipids in compositions which also contain neutral lipids, however, this is not necessary, because the emulsion not containing neutral lipids, are also suitable. The use of combined administration of phospholipids and neutral lipids is that of neutral lipids and phospholipids are associated to particles that resemble lipoprotei is s, but they differ in that they do not contain proteins or peptides, which, of course, always present in lipoproteins.

Most preferred is the use for the treatment compositions, in which the phospholipid is phosphatidylcholine, such as egg yolk phosphatidylcholine, soy phosphatidylcholine or sphingolipid. Of bile acids/salts of bile acids, the preferred choice is cholic acid and its salts, for example Holt sodium, deoxycholate sodium and chenodeoxycholic sodium. Of neutral lipids is preferable to use an ester of cholesterol or triglyceride, however, other neutral lipids, such as squalene or other hydrocarbon oils, di - and monoglycerides and antioxidants, such as vitamin E, may also be used.

Route of administration of the compositions may vary from intravenous bolus to other options, intravenous, depending on which one is most preferred. In that case, when the composition is administered intravenously bolus and contains triglycerides, special attention should be paid to the dose. It is well known that triglycerides are toxic, if you write them in too large quantities. However, the person skilled in the art can easily find the dose of the composition in such a way as to reduce or equityrelated the risk of poisoning triglycerides. In General, in the case of intravenous bolus injection, the composition should contain no more than 80% by weight of triglycerides or other neutral lipids, preferably not more than 70% by weight. More preferably, the composition for intravenous bolus injection should contain no more than 50% by weight of neutral lipids.

In that case, when the composition is introduced using other variants of intravenous injection, the risk of poisoning is greatly reduced. However, the ratios shown above are preferable to any form of intravenous injection, however, this requirement is not mandatory. Preferably, the dose ranges from 200 mg of bile acids/salts of bile acid or phospholipid per kg of body weight of the subject. It is also possible introduction dose component 800 mg/kg the dose can vary depending on the individual characteristics of the subject and the method of administration.

As described above, the compositions do not contain proteins and peptides, require at least one phospholipid or bile acids/salts of bile acids. As for phospholipids is preferable that the composition was attended by at least one neutral lipid, such as triglyceride, diglyceride or monoglyceride. The composition may also contain additional materials such as sterols (for example, cholesterol, beta-sitosterol), esterified or neeterificirovannah lipids (for example, esters of cholesterol or neeterificirovannah cholesterol), hydrocarbon oils, such as squalene, antioxidants, such as vitamin E, but this is not necessary. Of course, in the claimed compositions can be used not one, but several phospholipids or neutral lipids. In that case, when you use a combination of phospholipid and neutral lipid, the latter should be between 30% to 50% in weight relative to the total weight of the lipid in the composition.

Bile acids or bile salts can be used alone or in combination with phospholipid, neutral lipid, or both of these compounds. However, preferred is to use them in combination with the materials mentioned above. The optimal additional ingredients are also described above.

The present invention also relates to the use of the claimed compositions for the treatment of pathological conditions associated with infections caused by bacteria, as described above. These compositions preferably contain (in % by weight), approximately 5% to 30% of bile acids/salts of bile acids, approximately, from 3% to 50% of neutral lipids and approximately 10% to 95% phospholipids and do not contain proteins, Pat the Dov. Most preferred are compositions containing from 10-15% of bile acids/salts of bile acids, from 5% to 10% of neutral lipids, all the ingredients are balanced by the phospholipid. The term "does not contain protein or peptide"as used here, refers to compositions that do not contain sufficient amounts of proteins or peptides and used for the treatment of pathological conditions described above, but the proteins and peptides may be present in the compositions of trace residual amounts.

It should be noted that the presented percentages fair for compositions consisting of three components. In the case where the three-component composition is in combination with, for example, carrier, adjuvant or other optimal ingredients (e.g., described above), these percentages (of the total weight of the composition) will decrease; however, the proportion of each component in relation to each other will remain the same. It must be remembered that these therapeutic compositions are always almost completely free from proteins and peptides.

In the case where the composition does not contain bile acids or their salts and proteins, such free protein compositions preferably contain about 3% to 50% by weight of neutralcoloured, balance is at least one phospholipid. Preferably, the neutral lipid is a triglyceride, however, you can use any of the above-described neutral lipids. A preferred phospholipid is phosphatidylcholine.

Composition claimed in accordance with the present invention are effective for treatment or prevention of pathological conditions caused by Gram-positive bacteria, including (without limitation specified): sepsis syndrome, septic shock syndrome, systemic inflammatory response or "SIRS", SIRS with organ dysfunction or failure, organ dysfunction or failure, caused by Gram-positive bacteria.

Other possible aspects of the present invention are obvious to experts in this field and, therefore, not described in this specification.

It should be clear that the present invention examples are purely for illustrative purposes and in no way limits the scope of the invention; in addition, it is obvious that various aspects and embodiments of the invention can be partially or completely modified without changing its essence.

1. Pharmaceutical composition for the treatment and prevention of bacterial infections due to gram-positive bacteria, aderasa the effective number holonovel acid or salt holonovel acid, phosphatidylcholine and neutral lipids, in which neutral lipids and phospholipids associated in lipoproteinemia particles that do not contain proteins or peptides.

2. The composition according to claim 1, in which the neutral lipid is a triglyceride.

3. The composition according to claim 1, which contains from about 5 to 30% of the total weight of the composition holonovel acid or salt holonovel acid, from about 3 to 50% of the total weight of the composition of the neutral lipid, and from about 10 to 95% phosphatidylcholine.

4. The composition according to claim 1, in which salt holonovel acid is Halat sodium.

5. The composition according to claim 1, which is used in cases where gram-positive bacteria include Bacillus subtilis.

6. The composition according to claim 1, which is used in cases where the gram-positive bacterium produces lipoteichoic acid.

7. The composition according to claim 1, which is used in cases where a bacterial infectious disease caused by gram-positive bacteria selected from the group comprising sepsis, septic shock syndrome, systemic inflammatory response (SIRS), SIRS with organ dysfunction or failure, organ failure and organ dysfunction.



 

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18 cl, 4 ex, 4 tbl

FIELD: medicine; gastroenterology.

SUBSTANCE: after patient has been diagnosed with normal values of serum lipids, apolipoproteid B and balanced processes of lipid peroxidation (POL) and phosphatidyl serine (PS) that is less or equal to 10.4%, content of sphingomyelins (SP) from 24.1% to 27.6%, value of phosphatidyl ethanolamine (PE) equal or more than 29.2%, ratio of phosphatidyl choline PC/PE below 1.2, and also index of fatty acids unsaturation (IFAU) from 160.1 to 165.7, hypolipidemic dietotherapy (HD) is realised with enterosorbent hitamine or with taking of carbonated mineral water (CMW) and electrophoresis of mud extraction (EME) to liver area. If total cholesterol (TC) value is 5.0 -5.3 mM/l, cholesterol of low density lipoproteins (CLDL) is 2.9-3.3 mM/l and triglycerides make ≤0.9 mM/l, apolipoproteid B is ≤ 61.0 mg/dl, and (POL) lipid peroxidation processes are balanced with antioxidant protection (AOP) on the background of phosphatidyl serine content of 10.5%-13.6%, sphingomyelines content is ≤24.1% and IFAU makes 150.-160.0, HD is prescribed in complex with essential phospholipids (EP) or with enterosorbent hitamine with taking of HCMW and EME to liver area. If TC content is ≥5.4 mM/l, CLDL is ≥3.4 mM/l, CLDL is ≥1.7 mM/l, apolipoproteid B makes 112.0-183.0 mg/dl and total antioxidant activity of blood is ≤49% in combination of PC of 32.9%-33.4%, PE 27.6%-29.1%, ratio of PC/PE of 1.2 and IFAU is ≤150.4, HD is prescribed with inclusion of EP.

EFFECT: differentiated adequate pathogenetically oriented lipid-correcting therapy for this contingent of patients.

1 tbl, 1 ex, 1 dwg

FIELD: medicine.

SUBSTANCE: invention is related to cholesterol-6-O-acyl-β-D-galactopyranoside intended for inducing in a host mammal of immune reaction on B.burgdorferi, where (i) residual acyl represents a group recovered from organic fatty acids containing 1-25 carbon atoms; (ii) galactopyranoside ring structure optionally contains acyl group recovered from fatty acid in any position of the ring. The invention also covers mixed 3-O-(6-O-palmitoyl-(β-D-galactopyranosyl)-cholesterol and 3-O-(6-O-oleoyl-β-D-galactopyranosyl)cholesterol intended for inducing in a host mammal of immune reaction on B.Burgdorferi.

EFFECT: compounds are especially effective for prevention or treatment of Lim's disease.

7 cl, 4 tbl, 15 dwg, 15 ex

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