Synthetic intermediates for luteinizing hormone-releasing hormone (lhrh) antagonists, method for production thereof and method for production of lhrh antagonists

FIELD: biochemistry.

SUBSTANCE: invention relates to method for tripeptide production of formulae Ac-D-2Nal-D-4ClPhe-D-3Pal-OH and Boc-D-2Nal-D-4ClPhe-D-Pal-OH, which represent intermediates for synthesis of LHRH analogs in combination with acceptable heptapeptides in particular P1-Ser(P2)-NMeTyr(P3)-D-Lys(Nic)-Leu-Lys(iPr,P4)-Pro-D-AlaNH2 and P1-Ser(P2)-NMeTyr(P3)-D-Asn-Leu-Lys(iPr,P4)-Pro-D-AlaNH2 heptapeptides.

EFFECT: new synthetic intermediates for LHRH antagonists.

7 cl, 8 ex

 

The SCOPE of the INVENTION

The present invention relates to intermediate compounds for the synthesis of LHRH antagonists, the method of production of these intermediates and to a process for the preparation of LHRH antagonists.

PRIOR art

Releasing factor, luteinizing hormone (luteinizing hormone-releasing hormone, LHRH) controls the secretion of follicle-stimulating hormone (FSH) and luteinizing hormone (LH). The LHRH antagonists are compounds that can block the secretion of FSH and LH. They usually represent Nona - and Decapeptide (but can be shorter or longer), containing part or all of the structure of LHRH, where one or several amino acids are substituted by other natural amino acids and/or amino acids not found in nature.

Synthetic LHRH antagonists can be used for contraception and for the treatment of benign prostatic hyperplasia, gormonzavisimykh tumors of the breast and ovarian cancer, dysmenorrhea, endometriosis and other conditions. These synthetic LHRH antagonists have the General formula

where X represents a 5-6 natural and/or synthetic amino acid residues. More specifically they have the above-mentioned General formula, where X represents AA1-AA2-Leu-AA3-Pro-D-Ala, in particular, when AA1 is a PR is a native or synthetic amino acid and AA2 is a natural or synthetic amino acid or null AA3 is a natural or synthetic amino acid.

Although there are many methods of synthesis for the preparation of LHRH analogues, known from the prior art, there is a need for improvement, as the total output of LHRH analogues obtained using known methods, is low and, in addition, products may require time-consuming cleaning. In addition, the methods of synthesis of analogs of LHRH, known from the prior art, are very expensive.

The strategy of synthesis described in U.S. patent No. 5710246 to obtain Decapeptide or nonapeptide LHRH antagonists include a combination of intermediate compounds of Tripeptide representing amino acid residues from 1 to 3 (the countdown begins with N-Terminus of the peptide), with heptapeptide or Hexapeptide, respectively, representing amino acid residues 4-10 and 4-9, respectively. The intermediate connection Tripeptide described in US 5710246 And represents an ester of Boc-D-2Nal-D-4ClPhe-D-3Pal-O-Me or the corresponding benzyl or allyl ester.

TASKS INVENTIONS

Thus, the objective of the invention is to provide an intermediate connection Tripeptide for the synthesis of 3+7 and 3+6 LHRH analogues with improved yield and/or purity of the product.

Another objective of this invention is the fact that to suggest a method of obtaining such intermediate compounds of Tripeptide.

Another objective of this invention is to offer a method of receiving LHRH analogues, in which the combination of Tripeptide with hepta - or Hexapeptide.

Other objectives of this invention will become clear from the following summary of the invention, the description of the preferred embodiments and the accompanying claims.

DEFINITIONS AND ABBREVIATIONS

Regarding definitions and abbreviations used in this application, and which are generally accepted in the field of the invention, references are provided, in particular, on US 5710246 A.

SUMMARY of the INVENTION

In accordance with the present invention proposed a Tripeptide, representing amino acids 1-3 of LHRH antagonist, terminal amino group is protected with a Boc or Speakers and an end carboxyl group (i.e. terminal amino group No. 3) is not protected.

In accordance with this invention is described Tripeptide (I)

which is a useful intermediate compound in the synthesis method of the LHRH antagonist of General formula (II)

where X represents from 5 to 7 natural and/or synthetic amino acid residues, more preferably AA-AA2-Leu-AA3-Pro-D-Ala, in particular, where AA1 is a natural or synthetic amino acid and AA2 is a natural or synthetic amino acid or zero, AA3 represents a natural or synthetic amino acid.

Preferred is the use of Tripeptide (I) in the synthesis of the peptide of General formula (IIa)

where AA1 and AA2 have the meanings specified above, in particular the LHRH antagonist of the formula (III)

or even more preferably of the formula (IIIa)

In accordance commissioned by the invention are also described Tripeptide

have the same utility.

In addition, in accordance with this invention describes a method for Tripeptide formula (I)

or (IX)

at which carry out the following successive stages of production (I):

(a) interact Boc-D-4ClPhe-OH with HONSu with the formation of Boc-D-4ClPhe-OSu (VII);

(b) interact Boc-D-4ClPhe-OSu (VII) H-D-3Pal-HE with the formation of Boc-D-4ClPhe-D-3Pal-OH (VIII);

(C) interact Boc-D-4ClPhe-D-3Pal-OH (VIII) with Boc-D-2Nal-OSu, obtained by interaction of Boc-D-2Nal-OH with HONSu, with the formation of Boc-D-2Nal-D-4ClPhe-D-3Pal-OH (IX);

(g) carry out vzaimode the op perate Boc-D-2Nal-D-4ClPhe-D-3Pal-OH (IX) with acetic acid with the formation of Ac-D-2Nal-4ClPhe-D-3Pal-OH (I);

or successive stages (a) through (C) to obtain (IX).

When the method according to this invention for receiving LHRH antagonist exercise stage combination of Tripeptide (I) heptapeptide (IV) the General formula

where R4represents H or a protective group for amino, such as Boc, where AA1 and AA2 have the above significance, in particular, with heptapeptides (V) the General formula P1-Ser(P2)-NMeTyr(P3)-D-Lys(Nic)-Leu-Lys(iPr,P4)-Pro-D-AlaNH2(V), where R1selected from H or a protective group for amino, R2and R3independently selected from h, a protective group for IT, and R4has the above value, for receiving the LHRH antagonist Ac-D-2Nal-D-4ClPhe-D-3Pal-Ser-MeTyr-D-Lys(Nic)-Leu-Lys(iPr)-Pro-D-Ala-NH2(III)more specifically, with heptapeptides (Va) General formula P1-Ser(P2)-NMeTyr(P3)-D-Asn-Leu-Lys(iPr,P4)-Pro-D-AlaNH2(Va), where R1selected from H or a protective group for amino, R2and R3independently selected from h, a protective group for IT, and R4has the above value, for receiving the LHRH antagonist Ac-D-2Nal-D-4ClPhe-D-3Pal-Ser-MeTyr-D-Asn-Leu-Lys(iPr)-Pro-D-Ala-NH2(III).

Heptapeptide (V) described in US 5710246 A. Heptapeptide General formula (IV), including heptapeptide (Va)can be synthesized by standard modifications of the synthesis of (V) or by a combination of the appropriate BOC-amino acids is the apparatus for the synthesis of peptides (Beckman Model 990), as described in WO 94/40757, which also described the LHRH antagonist (III).

Alternatively, in the method according to this invention for receiving LHRH antagonist exercise stage combination of Tripeptide (IX)

with heptapeptides (IV) the General formula

where R1, R2, R4, AA1 and AA2 have the above values,

in particular, heptapeptides (V) the General formula

or even more preferably heptapeptide General formula (Va)

where R1selected from H or a protective group for amino, R2and R3independently selected from h, a protective group for IT, and R4has the above value, with subsequent replacement of the Boc group at the N-end acyl group, in particular acetyl group.

More specifically, heptapeptide General formula (V) represents heptapeptide (VI)

or even more preferably heptapeptide (VIa)

A special advantage of the method according to this invention is that instead of ester H-Pal-OR·HCl, can be used a cheaper starting material H-D-Pal-OH·2HCl; no need to remove the protective group from the original substances. So what Braz, the synthesis of this invention at one stage shorter and avoids loss of substance on the extra stage. Another advantage is that avoid formation of impurities on the stage of saponification. The formation of such impurities is well known. For example, the basic conditions at the stage of hydrolysis of ester cause partial racemization D-Pal. Another alternative to remove a group of ester prior art by catalytic hydrogenation (in the case of allyl or benzyl ester groups) causes the risk of loss of Cl from 4ClPhe, leading to the formation of Phe. Although the allyl group can be removed by the use of other reagents, complete removal is difficult to control.

The invention is explained in more detail hereinafter with reference to the preferred embodiment.

DESCRIPTION of the PREFERRED EMBODIMENT of the present INVENTION

Synthesis of Ac-D-2Nal-D-4ClPhe-D-3Pal-OH (I)

EXAMPLE 1. Boc-D-4ClPhe-OSu. Boc-D-4ClPhe-OH (299,75 g; 1.0 EQ.) and HONSu (184,1 g; 1.6 EQ.) dissolved in 2-propanol (4.5 l). The mixture is cooled to 0°and added DIC (164,1 g; 1.3 EQ.). The mixture is stirred for 16 hours, warming to room temperature. The product is filtered, washed with 2-propanol (1.5 l) and dried. Yield: 85%. Purity according to HPLC: 98.8 per cent.

EXAMPLE 2. Boc-D-4ClPhe-D-3Pal-OH. H-D-3Pal-OH, 2HCl (251,1 g; of 1.05 equiv.) and Boc-D-4ClPhe-OSu (396,8 g; 1.0 EQ.) dissolve Aut in dimethyl sulfoxide (DMSO) (3,33 l) and added NMM (318,8 g; 3,15 EQ.). The mixture is stirred for 16 hours at room temperature. Add 17 l of water and the pH adjusted to 4-4,5, which causes the precipitation of the product. The mixture is filtered and the product washed with water (3×5 l) to remove traces of DMSO, H-D-3Pal-OH and Boc-D-4ClPhe-OH. The product is dried. Yield: 80%. Purity according to HPLC: 97.8 per cent.

EXAMPLE 3. Boc-D-2Nal-OSu. Boc-D-2Nal-OH (315,4 g; 1.0 EQ.) dissolved in 2-propanol (6,8 l) at -10°and add IBC (157 g, 1.15 EQ.) and NMM (116 g; 1.15 EQ.). After stirring for 5-10 minutes add the mixture HONSu (230,1 g; 2.0 EQ.) in 2-propanol (1.4 l). Add a further quantity of NMM (10,1 g; 0.1 EQ.). After half an hour, add water (0,82 l) to dissolve the precipitated NMM·HCl. The product distinguish by filtration, washed with 2-propanol (1 l) and dried. Yield: 90%. Purity according to HPLC: 98.3%of.

EXAMPLE 4. Boc-D-Nal-D-4ClPhe-D-3Pal-OH.

(a) Remove the protection. Boc-D-4ClPhe-D-3Pal-OH (447, 93 g; 1.0 EQ.) dissolved in a mixture of ethyl acetate (3.4 l), acetic acid (675 ml) and MSA (454 ml, 7.0 EQ.) when 0°and left at this temperature for two hours. Add tetraethylammonium (tea) (1669 ml, 12 EQ.).

(b) Condensation. Boc-D-Nal-OSu (412,4 g; 1.0 EQ.) added to the neutralized mixture to remove protection at room temperature. The reaction mixture is left at this temperature for 2-4 hours. Add water 25% NH3(154 ml, 2.0 EQ.) to extinguish the remaining quantity hydroxysuccinimide ether was Added 1-butanol (4.5 l) to prevent the deposition of subsequent extraction.

(C) isolation and Purification. The reaction mixture is extracted twice at pH 6 (2×4.5 l of water) to remove the tea, at pH 9 (4.5 l of water)to remove MSA and, finally, at pH 7 (4.5 l of water). Extraction is carried out at 40-45°to prevent deposition. To the organic phase, add acetic acid (4.5 l; 1 vol.) and the mixture was concentrated in vacuo and evaporated in conjunction with acetic acid (4.5 l) to obtain a solid substance.

EXAMPLE 5. Ac-D-2Nal-D-4ClPhe-D-3Pal-ONa.

(a) Remove the protection. To solid substance Boc-D-2Nal-D-4ClPhe-D-3Pal-OH add water (90 ml), acetic acid (1.8 l) and MSA (454 ml; 7.0 EQ.) and the mixture is stirred for 1-2 hours at room temperature. The mixture is cooled to 0°With and neutralize the tea (1071 ml, 7.7 EQ.). The solution was concentrated in vacuo and evaporated twice together with toluene (2×2.5 l) to obtain the oil.

(b) Acetylation. Oil stage after removal of the protection is dissolved in toluene (2.0 l) and add acetyl and imidazol (132, 14 g). The mixture is stirred at room temperature for 1 hour and then add water (100 ml) to extinguish the remaining acetyl and midazolam.

(C) Cleaning. The mixture after acetylation is heated to 30-35°and add 1-butanol (4.5 l) to prevent deposition. The mixture is extracted twice at pH 5 (2×2.6 liters of water) and twice at pH 11 (2 x 2.6 l of water), using NaOH to bring the pH to 11. The last extracts add Aut methanol (2.25 liters) to prevent deposition. Add NaCl (130 g) when the first and last extraction in order to minimize the loss of product in the water phase.

(d) Allocation. To thoroughly mix the organic phase after extraction add heptane (15 l) and the resulting suspension allowed to mix at room temperature for at least 1 hour. The mixture is filtered and the product washed twice with heptane (2×3.5 l) and dried. Yield: 75% (from Boc-D-4ClPhe-D-3Pal-OH). Purity according to HPLC: 92%. Analysis of amino acids: 2Nal: 1,1; 4ClPhe: 1,0; 3Pal: 0,9. Mass spectrum (MS): mass 586. Na: 4,6%.

EXAMPLE 6. Ac-D-2Nal-D-4ClPhe-D-3Pal-OH·DCHA

(a) Remove the protection. To solid substance Boc-D-2Nal-D-4ClPhe-D-3Pal-OH add water (90 ml), acetic acid (1.8 l) and MSA (454 ml; 7.0 EQ.) and the mixture is stirred for 1-2 hours at room temperature. The mixture is cooled to 0°With and neutralize the tea (1071 ml, 7.7 EQ.). The solution was concentrated in vacuo and together twice evaporated with toluene (2×2.5 l) to obtain the oil.

(b) Acetylation. Oil after removal of the protection is dissolved in toluene (2.0 l) and add acetylimidazole (132,14 g). The mixture is stirred at room temperature for 1 hour, followed by addition of water (100 ml) to absorb the remaining acetylimidazole.

(C) Cleaning. The mixture is heated to 30-35°and add 1-butanol (4.5 l) to prevent deposition. The mixture twice extragear the Ute at pH 7 (2× 2.6 liters of water), once at pH 9-9,5 (2.6 liters of water) and once at pH 7 (2.6 liters of water). Add dicyclohexylamine (DCHA) and the mixture was concentrated in vacuo. The product is suspended in 1-butanol (4.5 l) at 50°and added slowly to thoroughly mix the heptane (27 l). The mixture was stirred at 0°C overnight, filtered and the product washed twice with a mixture of 1-butanol/heptane (1:3; 2×4.8 l) and twice with heptane (2×4.5 l). Yield: 65% (from Boc-D-4ClPhe-D-3Pal-OH). Purity according to HPLC: 94,2%. Analysis of amino acids: 2Nal: 1,1; 4ClPhe: 1,0; 3Pal: 0,9. MS: mass 586 (free peptide).

EXAMPLE 7. Ac-D-2Nal-D-4ClPhe-D-3Pal-OH

(a) Remove the protection. To a solution of solids Boc-D-2Nal-D-4ClPhe-D-3Pal-OH add water (90 ml), acetic acid (1.8 l) and MSA (454 ml; 7.0 EQ.) and the mixture is stirred for 1-2 hours at room temperature. The mixture is cooled to 0°With and neutralize the tea (1071 ml, 7.7 EQ.). The solution was concentrated in vacuo and evaporated twice together with toluene (2×2.5 l) to obtain the oil.

(b) Acetylation. Oil after removal of the protection is dissolved in toluene (2.0 l) and add acetylimidazole (132,14 g). The mixture is stirred at room temperature for 1 hour and then add water (100 ml) to extinguish the remaining acetyl and midazolam.

(C) Cleaning. The mixture after acetylation is heated to 30-35°and add 1-butanol (4.5 l) to prevent deposition. The mixture twice extragroup pH 7 (2× 2.6 liters of water), once at pH 9-9,5 (2.6 liters of water) and once at pH 7 (2.6 liters of water). The mixture was concentrated in vacuo to obtain an oil which is dissolved in acetic acid (750 ml), concentrated, again dissolved in acetic acid (750 ml) and added slowly to a stirring mixture of heptane/ethyl acetate (3:1; 3,6 l). The mixture is left to mix at 0°With during the night. The mixture is filtered and the product washed twice with a mixture of ethyl acetate/heptane (1:3; 2×3,6 l) and twice with heptane (2×3,6 l). Yield: 70% (from Boc-D-4ClPhe-D-3Pal-OH). Purity according to HPLC: 93.9 per cent. Analysis of amino acids: Nal; 1,1; 4ClPhe: 1,0; 3Pal: 0,9. MS: mass 586 (free peptide).

EXAMPLE 8. Ac-D-2Nal-D-3Pal-D-4ClPhe-Ser(tBu)-MeTyr-D-Asn-Leu-Lys(iPr,Boc)-Pro-D-Ala-NH2

Z-Ser(tBu)-MeTyr(Bzl)-D-Asn-Leu-Lys(iPr,Boc)-Pro-D-Ala-NH2(122,8 g) dissolved in ethanol (1.2 l) and add water (400 ml). Hydrogenation initiated by adding 5% Pd/C (18,4 g) and hydrogen bubbled through the reaction vessel. Continuous addition of HCI (aq.) keep a constant pH value of 2.5-3.0 this mixture. After 1 hour of Pd/C is removed by filtration, and the pH of the filtrate is brought to a value of 3.6 by addition of NaOH (aq.). The filtrate is then evaporated, to the residue is added ethanol (360 ml) and again carry out the process of evaporation. This procedure is repeated once with ethanol (360 ml) and once with ethyl acetate (360 ml).

The residue is suspended in ethyl acetate (1.25 l) and added to a solution of AC-D-2Nal-D-3Pal-D-4ClPhe-OH, DCHA (dicyclo Xiumin) (60,9 g) in DMSO (260 ml). HOBt (1-hydroxybenzotriazole) (15.3 g) and CuCl2·N2(8.5 g) is added to the solution and then cooled to 0-5°C.

DCC (dicyclohexylcarbodiimide) (41,3 g) dissolved in ethyl acetate (33 ml) and added to a solution of the peptide with the speed at which the temperature is maintained below 10°C. After the addition the reaction mixture was kept at 0-5°C for 44 hours. Add water (77 ml), and this reaction mixture is stirred for 30 minutes and filtered. DCU-filter residue is washed with ethyl acetate (2×60 ml).

The combined organic phases are then extracted three times. First, water is added, and the pH adjusted to a value of 9.1 by adding NaOH (aq.). After stirring for 30 min add 25% NH3(aq.) (46.2 ml)and the phases provide an opportunity to separate. The aqueous phase is discarded and the organic phase extracted with water (1,54 l) and 25% N (aq.) (23.1 ml). After stirring for 30 minutes the phases are separated, and the aqueous phase discarded. The organic phase is finally extracted with water (1,54 l). After separation of the phases the aqueous phase is discarded.

The organic phase is evaporated on a rotary evaporator, and the residue is dissolved in acetone (1,54 l) at 40°and added to heptane (460 ml), preheated to 40°C. the Mixture is cooled to 4-8°and leave to stand for 18 hours. Fallen in sediment produktvideo filtering and the residue on the filter is washed twice with a mixture of acetone (208 ml) and heptane (92 ml), previously cooled to 2°C. the Product is then dried in a vacuum Cabinet.

The dry product twice subjected to recrystallization according to the following procedure: the product is dissolved in ethanol and evaporated on a rotary evaporator. The residue is then dissolved in acetone, is added to the heptane and cooled, filtered and washed according to the procedure described above. Yield 84 g (53,4%).

1. The method of obtaining Tripeptide, including its salt of the formula (I)

or (IX)

,

at which sequentially perform the following stage of obtaining the compound (I):

(a) interact Boc-D-4ClPhe-OH with HONSu education;

(b) interactH-D-3Pal-OH with education;

(V) remove protectionand interact with Boc-D-2Nal-OSu obtained by reacting Boc-D-2Nal-OH with HONSu, with formation;

(g) carry out interactionwith acetic acid and then acetimidoyl education;

or serial the stage performance communications with (a) (b) to obtain (IX).

2. The method of obtaining antagonist releasing factor, luteinizing hormone (LHRH) or its pharmaceutically acceptable salt, at which stage of the method according to claim 1 getting Tripeptideand then communicate the received Tripeptide with heptapeptides (IV) the General formula

where R1selected from H or a protective group for amino, R2represents H or a protective group for HE, R4represents H or a protective group for amino, such as Boc, AA1 is a natural or synthetic amino acid, AA2 represents a natural or synthetic amino acid or is absent.

3. The method according to claim 2, where heptapeptide General formula (IV) represents heptapeptide General formula

where R3represents H or a protective group for IT.

4. The method according to claim 2, where heptapeptide General formula (IV) represents heptapeptide General formula

where R3represents H or a protective group for IT.

5. The method according to claim 3, where heptapeptide General formula (V) represents heptapeptide formula

6. The method according to claim 4, where heptapeptide formula (VI) p is ecstasy heptapeptide formula

7. The Tripeptideor its salt obtained by the method according to claim 1.



 

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SUBSTANCE: invention proposes peptide of the structure Tyr-Ser-Leu and a pharmaceutical composition based on thereof that is used for stimulating antitumor immune response. Also, invention proposes methods for treatment of mammal and for modulation of the immune response. Proposed inventions expand assortment of agents used in treatment of cancer diseases.

EFFECT: valuable medicinal properties of peptide and pharmaceutical composition.

20 cl, 48 tbl

FIELD: synthesis of biologically active compounds.

SUBSTANCE: invention provides 1,5-benzothiazepines of general formula I (formulae presented below), in which Rv and Rw are independently selected from hydrogen and C1-C5-alkyl; one of Rx and Ry represents hydrogen or C1-C6-alkyl and the other hydroxy or C1-C6-alkoxy; Rz is selected from halogen, nitro, cyano, hydroxy, amino, carboxy, carbamoyl, mercapto, sulfamoyl, C1-C6-alkyl, and other residues indicated in claim 1 of invention; v is a number from 0 to 5; one of R4 and R5 represents group of general formula IA; R3 and R6 and the second from R4 and R5 are independently selected from hydrogen, halogen, nitro, cyano, hydroxy, amino, carboxy, carbamoyl, mercapto, sulfamoyl, C1-C6-alkyl, and other residues indicated in claim 1; R3 and R6 and the second from R4 and R5 being optionally substituted by one or several R16 groups at their carbon atoms; D represents -O-, -N(Ra)-, -S(O)b- or -CH(Ra)-, wherein Ra is hydrogen or C1-C6-alkyl; and b=0-2; ring A represents aryl or heteroaryl and is optionally substituted by one or several substituents selected from R17; R7 represents hydrogen, C1-C4-alkyl, carbocyclyl, or heterocyclyl and is optionally substituted by one or several substituents selected from R18; R8 represents hydrogen or C1-C4-alkyl; R9 represents hydrogen or C1-C4-alkyl; R10 represents hydrogen or C1-C4-alkyl, carbocyclyl, or heterocyclyl and is optionally substituted by one or several substituents selected from R19; R11 represents carboxy, sulfo, sulfino, phosphono, tetrazolyl, -P(O)(ORc)(ORd), -P(O)(OH)(ORc), -P(O)(OH)(Rd), or -(O)(ORc)(Rd), wherein Rc and Rd are independently selected from C1-C6-alkyl; or R11 represents group of general formula IB, in which X is -N(Rq)-, N(Rq)C(O)-, -O-, or -S(O)a, wherein a=0-2; and Rq is hydrogen or C1-C4-alkyl; R12 represents hydrogen or C1-C4-alkyl; R13 and R14 are independently selected from hydrogen, C1-C4-alkyl, carbocyclyl, heterocyclyl, or R23, of which C1-C4-alkyl, carbocyclyl, heterocyclyl, or R23 can be optionally independently substituted by one or several substituents selected from R20; R15 represents carboxy, sulfo, sulfino, phosphono, tetrazolyl, -P(O)(ORe)(ORf), -P(O)(OH)(ORe), -P(O)(OH)(Re), or -P(O)(ORe)(Rf), wherein Re and Rf are independently selected from C1-C6-alkyl; or R15 represents group of general formula IC, in which R24 is selected from hydrogen and C1-C4-alkyl; R24 is selected from hydrogen, C1-C4-alkyl carbocyclyl, heterocyclyl, and R27, of which C1-C4-alkyl, carbocyclyl, heterocyclyl, or R27 can be optionally independently substituted by one or several substituents selected from R28; R26 is selected from carboxy, sulfo, sulfino, phosphono, tetrazolyl, -P(O)(ORg)(ORh), -P(O)(OH)(ORg), -P(O)(OH)(Rg), or -P(O)(ORg)(Rh), wherein Rg and Rg are independently selected from C1-C6-alkyl; p=1-3; wherein meanings for R13 can be the same or different; q=0-1; r=0-3; wherein meanings for R14 can be the same or different; m=0-2; wherein meanings for R10 can be the same or different; n=1-3; wherein meanings for R7 can be the same or different; z=0-3; wherein meanings for R25 can be the same or different; R16, R17, and R18 are independently selected from halogen, nitro, cyano, hydroxy, carbamoyl, mercapto, sulfamoyl, C1-C4-alkyl, C2-C4-alkenyl, C2-C4-alkynyl, C1-C4-alkoxy, C1-C4-alkanoyl, C1-C4-alkanoyloxy, N-(C1-C4-alkyl)amino, N,N-(di-C1-C4-alkyl)amino, C1-C4-alkyl-S(O)a (wherein a=0-2), C1-C4-alkoxycarbonyl, N-(C1-C4-alkyl)sulfamoyl, and N,N-(di-C1-C4-alkyl)sulfamoyl; wherein R16, R17, and R18 can be optionally independently substituted by one or several of R21 at their carbon atoms; R19, R20, R23, R27, and R28 are independently selected from halogen, nitro, cyano, hydroxy, carbamoyl, mercapto, sulfamoyl, C1-C4-alkyl, C2-C4-alkenyl, C2-C4-alkynyl, C1-C4-alkoxy, C1-C4-alkanoyl, C1-C4-alkanoyloxy, N-(C1-C4-alkyl)amino, N.N-(di-C1-C4-alkyl)amino, C1-C4-alkanoylamino, N-(C1-C4-alkyl)carbamoyl, N,N-(di-C1-C4-alkyl)carbamoyl, C1-C4-alkyl-S(O)a (wherein a=0-2), C1-C4-alkoxycarbonyl, N-(C1-C4-alkyl)sulfamoyl, N,N-(di-C1-C4-alkyl)sulfamoyl, carbocyclyl, heterocyclyl, sulfo, sulfino, amidino, phosphono, -P(O)(ORa)(ORb), -P(O)(OH)(ORa), -P(O)(OH)(Ra), or -P(O)(ORa)(Rb), wherein Ra and Rb are independently selected from C1-C6-alkyl and wherein R19, R20, R23, R27, and R28 can be optionally independently substituted by one or several of R22 at their carbon atoms; R21 and R22 are independently selected from halogen, hydroxy, cyano, carbamoyl, mercapto, sulfamoyl, trifluoromethyl, trifluoromethoxy, methyl, ethyl, methoxy, ethoxy, vinyl, allyl, ethynyl, methoxycarbonyl, formyl, acetyl, formamido, acetylamino, acetoxy, methylamino, dimethylamino, N-methylcarbamoyl, N,N-dimethylcarbamoyl, methylthio, methylsulfinyl, mesyl, N-methylsulfamoyl, N,N-dimethylsulfamoyl; or pharmaceutically acceptable salt thereof, solvate, or salt solvate. Described are also method for preparing compounds of formula I, pharmaceutical compositions based on compounds I, and a method for achieving inhibiting effect relative to interscapular brown adipose tissue (IBAT), and intermediates. (I), (IA), (IB), (IC).

EFFECT: expanded synthetic possibilities in the 1,5-benzothiazepine series.

36 cl, 121 ex

The invention relates to a series peptidergic heterocyclic compounds, intermediates used in their receiving and containing pharmaceutical compositions

FIELD: medicine.

SUBSTANCE: invention relates to high stable pharmaceutical component containing peptide of human parathyroid hormone or derivative thereof and acetic acid in amount of less than chemical equivalent in relates to peptide of human parathyroid hormone or derivative thereof. Due to low acetic acid level obtained pharmaceutical compositions have high stability and are comfortable in application.

EFFECT: nasal compositions with increased stability.

The invention relates to novel antagonists of LHRH and complexes between the antagonists and VВ12

The invention relates to peptide compounds, namely the Decapeptide of formula I: x-X1-X2-X3-Ser-Tyr-X6-Leu-X8-Pro-X10(I), where X is acyl; X1=D-Nal; X2=D-Cpa; X3=D-Рal; X6=D-Cit, D-Hei, D-Orn, D-Lys, D-Neu; X8= Orn, Arg, Lys, L-Neu ; X10= D-AlaNH2, Neu is a residue of formula IX or IV; Z is a group-O - or-S-, n = 1-6

FIELD: medicine.

SUBSTANCE: invention relates to high stable pharmaceutical component containing peptide of human parathyroid hormone or derivative thereof and acetic acid in amount of less than chemical equivalent in relates to peptide of human parathyroid hormone or derivative thereof. Due to low acetic acid level obtained pharmaceutical compositions have high stability and are comfortable in application.

EFFECT: nasal compositions with increased stability.

FIELD: biochemistry.

SUBSTANCE: invention relates to method for tripeptide production of formulae Ac-D-2Nal-D-4ClPhe-D-3Pal-OH and Boc-D-2Nal-D-4ClPhe-D-Pal-OH, which represent intermediates for synthesis of LHRH analogs in combination with acceptable heptapeptides in particular P1-Ser(P2)-NMeTyr(P3)-D-Lys(Nic)-Leu-Lys(iPr,P4)-Pro-D-AlaNH2 and P1-Ser(P2)-NMeTyr(P3)-D-Asn-Leu-Lys(iPr,P4)-Pro-D-AlaNH2 heptapeptides.

EFFECT: new synthetic intermediates for LHRH antagonists.

7 cl, 8 ex

FIELD: chemistry.

SUBSTANCE: invention concerns compounds of the formula M-N-O-P-G, where M is a metal-chelating agent (in a complex with a radioactive metal nuclide or not), N-O-P is a linker, and G is a peptide for target transportation to a GRP receptor. Also the invention concerns methods of visualisation for a patient with the help of the claimed compounds, methods of obtaining a diagnostic visualisation and radiotherapeutic medium.

EFFECT: improved compounds for application in diagnostics visualisation or therapy.

49 cl, 51 dwg, 8 tbl, 58 ex

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology, precisely to cellular biology and experimental oncology and can be used for development of new methods aimed at antitumorigenesis. Human chorionic gonadotropin (HCG) is used for stimulation of pathological process and disorganisation of cell carcinoma culture of throat Hep-2.

EFFECT: invention allows effecting turmoral Hep-2 cell lines, excluding lymphocytic mediation, as well as hormone effect through the specific receptors with having signs of tumor degradation in the condition of cell culture.

2 dwg, 2 tbl, 2 ex

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology, in particular to production of hormones and can be used for culturing invertebrates. Gonadotrophin, selected from the invertebrate Asterina pectinifera is a peptide with molecular weight 4500-4900, it has two subunits, the protein structure of which is combined with SS-bridges, formed between residues of SH cysteine contained in the subunits.

EFFECT: interfusion and oxidation of these two subunits after synthesis allow producing gonadotrophin having gonadal promoting activity.

4 cl, 8 dwg, 1 tbl, 2 ex

FIELD: medicine.

SUBSTANCE: invention concerns area of medicine and concerns compositions and medicinal forms on the basis of Gastrinum, application and reception methods. The essence of the invention includes the bond of Gastrinum representing conjugates of fragments of amino-acid sequence of Gastrinum, possessing functional ability to contact Gastrinum/SSK receptor, with various carriers, including application amino-acid spacers, and application of bifunctional sewing agents, and also a method of treatment by the compositions including bond of Gastrinum, sick of diabetes.

EFFECT: advantage of the invention consists in action prolongation.

7 cl, 8 ex, 3 tbl, 2 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to conjugates of Disorazol with cell adhesion molecules, such as peptides, proteins, hormones, blood proteins, and methods for preparing conjugates.

EFFECT: conjugates of Disorazol may be used as drug preparations for treating various tumours.

19 cl, 17 dwg, 2 tbl, 17 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a diagnostic and therapeutic agent which is a bombesin analogue peptide antagonist conjugate having the general formula (I), [A-(B)n]x-C, wherein A represents a metal chelator containing at least one radionuclide metal, B represents a spacer bound to N terminal C, or a covalent bond, and C represents a bombasin analogue peptide antagonist, wherein additionally x represents an integer 1 to 3, and n represents an integer 1 to 6.

EFFECT: preparing the therapeutic agent representing the bombesin analogue peptide antagonist conjugate.

17 cl, 12 dwg, 1 tbl, 13 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to biochemistry, particularly to a recombinant human follicle-stimulating hormone (FSH). The above recombinant human FSH contains α2,3- and α2,6-cyalilation, wherein 10% to 90% of the total syalilation represents α2,3-syalilation, and contains cyalic acid in an amount (expressed as a relation of moles of syalic acid to moles of the protein) of 6 mole/mole to 15 mole/mole. What is also declared is a pharmaceutical composition for treating infertility containing the above recombinant human FSH.

EFFECT: invention enables improving pharmacokinetic properties of the recombinant FSH.

12 cl, 13 dwg, 11 ex

FIELD: biotechnology; medicine.

SUBSTANCE: invention relates to field of biotechnology and medicine. Disclosed is a pharmaceutical composition having human chorionic gonadotropin (hCG) activity. Composition comprises recombinant hCG, comprising α2,3-linked sialic acid and α2,6-linked sialic acid, and includes mono (1S), di (2S), tri (3S) and tetra (4S) sialylated structures, and a pharmaceutically acceptable carrier.

EFFECT: composition has high activity, which is equal to 27477 IU hCG/mg, and recombinant hCG has a sialic acid content of 15 mol/mol protein to 25 mol/mol protein.

13 cl, 6 dwg, 11 ex

FIELD: synthesis of biologically active compounds.

SUBSTANCE: invention provides 1,5-benzothiazepines of general formula I (formulae presented below), in which Rv and Rw are independently selected from hydrogen and C1-C5-alkyl; one of Rx and Ry represents hydrogen or C1-C6-alkyl and the other hydroxy or C1-C6-alkoxy; Rz is selected from halogen, nitro, cyano, hydroxy, amino, carboxy, carbamoyl, mercapto, sulfamoyl, C1-C6-alkyl, and other residues indicated in claim 1 of invention; v is a number from 0 to 5; one of R4 and R5 represents group of general formula IA; R3 and R6 and the second from R4 and R5 are independently selected from hydrogen, halogen, nitro, cyano, hydroxy, amino, carboxy, carbamoyl, mercapto, sulfamoyl, C1-C6-alkyl, and other residues indicated in claim 1; R3 and R6 and the second from R4 and R5 being optionally substituted by one or several R16 groups at their carbon atoms; D represents -O-, -N(Ra)-, -S(O)b- or -CH(Ra)-, wherein Ra is hydrogen or C1-C6-alkyl; and b=0-2; ring A represents aryl or heteroaryl and is optionally substituted by one or several substituents selected from R17; R7 represents hydrogen, C1-C4-alkyl, carbocyclyl, or heterocyclyl and is optionally substituted by one or several substituents selected from R18; R8 represents hydrogen or C1-C4-alkyl; R9 represents hydrogen or C1-C4-alkyl; R10 represents hydrogen or C1-C4-alkyl, carbocyclyl, or heterocyclyl and is optionally substituted by one or several substituents selected from R19; R11 represents carboxy, sulfo, sulfino, phosphono, tetrazolyl, -P(O)(ORc)(ORd), -P(O)(OH)(ORc), -P(O)(OH)(Rd), or -(O)(ORc)(Rd), wherein Rc and Rd are independently selected from C1-C6-alkyl; or R11 represents group of general formula IB, in which X is -N(Rq)-, N(Rq)C(O)-, -O-, or -S(O)a, wherein a=0-2; and Rq is hydrogen or C1-C4-alkyl; R12 represents hydrogen or C1-C4-alkyl; R13 and R14 are independently selected from hydrogen, C1-C4-alkyl, carbocyclyl, heterocyclyl, or R23, of which C1-C4-alkyl, carbocyclyl, heterocyclyl, or R23 can be optionally independently substituted by one or several substituents selected from R20; R15 represents carboxy, sulfo, sulfino, phosphono, tetrazolyl, -P(O)(ORe)(ORf), -P(O)(OH)(ORe), -P(O)(OH)(Re), or -P(O)(ORe)(Rf), wherein Re and Rf are independently selected from C1-C6-alkyl; or R15 represents group of general formula IC, in which R24 is selected from hydrogen and C1-C4-alkyl; R24 is selected from hydrogen, C1-C4-alkyl carbocyclyl, heterocyclyl, and R27, of which C1-C4-alkyl, carbocyclyl, heterocyclyl, or R27 can be optionally independently substituted by one or several substituents selected from R28; R26 is selected from carboxy, sulfo, sulfino, phosphono, tetrazolyl, -P(O)(ORg)(ORh), -P(O)(OH)(ORg), -P(O)(OH)(Rg), or -P(O)(ORg)(Rh), wherein Rg and Rg are independently selected from C1-C6-alkyl; p=1-3; wherein meanings for R13 can be the same or different; q=0-1; r=0-3; wherein meanings for R14 can be the same or different; m=0-2; wherein meanings for R10 can be the same or different; n=1-3; wherein meanings for R7 can be the same or different; z=0-3; wherein meanings for R25 can be the same or different; R16, R17, and R18 are independently selected from halogen, nitro, cyano, hydroxy, carbamoyl, mercapto, sulfamoyl, C1-C4-alkyl, C2-C4-alkenyl, C2-C4-alkynyl, C1-C4-alkoxy, C1-C4-alkanoyl, C1-C4-alkanoyloxy, N-(C1-C4-alkyl)amino, N,N-(di-C1-C4-alkyl)amino, C1-C4-alkyl-S(O)a (wherein a=0-2), C1-C4-alkoxycarbonyl, N-(C1-C4-alkyl)sulfamoyl, and N,N-(di-C1-C4-alkyl)sulfamoyl; wherein R16, R17, and R18 can be optionally independently substituted by one or several of R21 at their carbon atoms; R19, R20, R23, R27, and R28 are independently selected from halogen, nitro, cyano, hydroxy, carbamoyl, mercapto, sulfamoyl, C1-C4-alkyl, C2-C4-alkenyl, C2-C4-alkynyl, C1-C4-alkoxy, C1-C4-alkanoyl, C1-C4-alkanoyloxy, N-(C1-C4-alkyl)amino, N.N-(di-C1-C4-alkyl)amino, C1-C4-alkanoylamino, N-(C1-C4-alkyl)carbamoyl, N,N-(di-C1-C4-alkyl)carbamoyl, C1-C4-alkyl-S(O)a (wherein a=0-2), C1-C4-alkoxycarbonyl, N-(C1-C4-alkyl)sulfamoyl, N,N-(di-C1-C4-alkyl)sulfamoyl, carbocyclyl, heterocyclyl, sulfo, sulfino, amidino, phosphono, -P(O)(ORa)(ORb), -P(O)(OH)(ORa), -P(O)(OH)(Ra), or -P(O)(ORa)(Rb), wherein Ra and Rb are independently selected from C1-C6-alkyl and wherein R19, R20, R23, R27, and R28 can be optionally independently substituted by one or several of R22 at their carbon atoms; R21 and R22 are independently selected from halogen, hydroxy, cyano, carbamoyl, mercapto, sulfamoyl, trifluoromethyl, trifluoromethoxy, methyl, ethyl, methoxy, ethoxy, vinyl, allyl, ethynyl, methoxycarbonyl, formyl, acetyl, formamido, acetylamino, acetoxy, methylamino, dimethylamino, N-methylcarbamoyl, N,N-dimethylcarbamoyl, methylthio, methylsulfinyl, mesyl, N-methylsulfamoyl, N,N-dimethylsulfamoyl; or pharmaceutically acceptable salt thereof, solvate, or salt solvate. Described are also method for preparing compounds of formula I, pharmaceutical compositions based on compounds I, and a method for achieving inhibiting effect relative to interscapular brown adipose tissue (IBAT), and intermediates. (I), (IA), (IB), (IC).

EFFECT: expanded synthetic possibilities in the 1,5-benzothiazepine series.

36 cl, 121 ex

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