Ionic molecular conjugates biodegradable polyesters and bioactive polypeptides

 

The invention relates to a complex of the polyester containing one or more free carboxyl groups and characterized by the ratio of carboxyl and hydroxyl groups greater than one. This complex polyester is a copolymer-caprolactone and glycolide at a ratio (90-99):(1-10) and obtained in the presence of initiator is citric acid. The invention relates to a variant of the complex polyester and variants of a composition containing a complex of the polyester. The invention allows to obtain compositions which are chemically linked biocompatible, biodegradable complex polyester with oligopeptides, polypeptides, peptides and/or proteins in the form of a homogeneous ion product. In addition, compositions of the invention are easily optimized for the acquisition of functional properties for a large load of therapeutically active polypeptide. 6 N. and 18 C.p. f-crystals, 6 tab., 3 Il.

Reference to related applications

This application is a partial continuation of concurrently pending application No. 08/867308, filed June 2, 1997, which granted U.S. patent No. 5863985 January 26, 1999, which is a partial continuation of application No. 08/464735, filed 29 the s Phase of PCT/US 94/00148, submitted January 5, 1994, which is the PCT phase application Ireland No. 930005, filed January 6, 1993

Background of invention

This invention relates to a continuous release of biologically active polypeptides.

Numerous systems of drug delivery was developed, studied and used for controlled release in vivo pharmaceutical compositions. For example, polyesters, such as poly(DL-lactic acid), poly(glycolic acid), poly(-caprolacton) and various other copolymers used for the release of biologically active molecules, such as progesterone; such compounds have the form of microcapsules, films or sticks (Pitt CG, Marks TA and A. Schindler 1980). During implantation of the composition of the polymer/therapeutic agent, for example, subcutaneously or intramuscularly at therapeutic agent is released over a certain period of time. Such biocompatible, biodegradable polymer systems are designed to allow the prisoner in the composition of therapeutic agent to diffuse from the polymer matrix. With the release of therapeutic agent, the polymer degrades in vivo, thus avoiding chirurgiche such degradation of the polymers can be regulated by the availability of essential links for non-enzymatic autocatalytic hydrolysis of the polymer components.

Some publications EPO and patents U.S. dedicated to the design of polymer matrix and its role in the regulation of the rate and extent of release of therapeutic agent in vivo.

For example, Deluca (EPO publication 0467389 A2/Univ of Kentucky) described the physical interaction between hydrophobic biodegradiruemym the polymer and the protein or polypeptide. The resulting composition was a mixture of therapeutic agent and a hydrophobic polymer that has kept the diffusive release of therapeutic agent from the matrix after the introduction of the subject.

Hutchison (U.S. patent 4767628/ICI) observed the release of therapeutic agent through a uniform distribution in the polymeric device. Found that this arrangement provides an adjustable continuous release through the overlap of two phases: the first, the dependent diffusion leaching of drug from the surface of the composition, and the second induced degradation of polymer release water channels.

Summary of invention

In General terms, the invention presents the pharmaceutical composition of continuous release, consisting of a complex of the polyester containing free COOH groups, ion paired with the biological the th least 50 wt.% polypeptide present in the composition, are ion conjugated with complex polyester.

In preferred embodiments, the implementation of complex polyester modified so as to increase the ratio of end groups, carboxyl to hydroxyl, from more than one to approaching infinity, that is, all of the hydroxyl groups may be substituted by carboxy. Examples of suitable polyesters are polyesters, derivatives of compounds such as L-lactic acid, D-lactic acid, DL-lactic acid,-caprolactone, p-dioxanone,-hexanoic acid, substituted and unsubstituted trimethylantimony (TMS), 1,5-dioxan-2-it, 1,4-dioxan-2-it, glycolide, glycolic acid, L-lactide, D-lactide, DL-lactide, mesolectal, oxalate alkylene, oxalate cycloalkene, succinate of alkylene, (-hydroxybutyrate) and optically active isomers, racemates or copolymers of any of the above, which is substituted by TMS substituted (C1-C4)alkyl, preferably the stands. Can also be used for other polymers with heterotopia related traditional complex polyesters (for example, polyarteritis, beep with malic acid, citric acid or tartaric acid.

In preferred embodiments, the implementation of complex polyester partially contains terminal acidic residues at the expense of glutaric anhydride. In other preferred embodiments, the complex polyester contains terminal acidic residues at the expense of glutaric anhydride. Preferably complex polyester is characterized by an average degree of polymerization between 10 and 300, and more preferably between 20 and 50.

Ionic molecular conjugates according to the invention is preferably produced from polyesters with terminal residues of polycarboxylic acids, conjugated with monobasic and polybasic biologically active polypeptides containing at least one effective ionogenic amine group. Alternative any complex polyester can be used for the formation of ionic molecular conjugate according to the invention, provided that he has previously been treated with a suitable base, for example NaOH. In addition, there may be used any cyclotosaurus peptide, such as peptide release growth hormone (GHRP), the hormone that stimulates luteinizing hormone (LHRH), somatostatin, bombezin, peptide, gastrin releasing (GRP), calcitonin, who, secretin, parathyroid hormone (PTH), enkephalin, endothelin, a peptide, releasing calcitonin gene (CGRP), neuromedin, protein, related to parathyroid hormone (Rtrr), glucagon, neurotensin, adrenocorticotropic hormone (ACTH), peptide YY (PYY), a peptide that stimulates glucagon (GLP), vasoactive putting peptide (VIP), peptide, activates the pituitary adenylate cyclase (RASAR), motilin, substance P, neuropeptide Y (NPY), TSH (thyroid stimulating hormone) and analogues and fragments. Such ionic molecular conjugates capable of release in vivo bioactive components with a predetermined speed set according to the chemical structure, molecular weight and pKa of both components of the resulting conjugates. The mechanism of release of the drug causes the transformation of insoluble forms of the conjugate to components, soluble in water, partly by hydrolysis of the hydrophobic complex of the polyester. Thus, release of the bioactive polypeptide independently increases with (a) the reduction of the difference in pKa of the bioactive polypeptide and a complex of the polyester, (b) chemical reactivity of the chain complex of the polyester, which is reflected in nucleoplasty carbonyl, (C) the reduction of density polyester that is related is preferred directions polypeptide comprises 1 to 50 wt.% from the total mass of ionic molecular conjugate, and more preferably 85%, more preferably 95% and most preferably 99% of the polypeptide present in the composition, ion is conjugated with a complex polyester; polyester component ionic molecular conjugate is characterized by a viscosity of from about 0.05 to 0.7 DL/g in chloroform; the average molecular weight of the polyester is about 1200-40000.

Polymeric ionic molecular conjugates of the invention can be easily obtained in the injectable microspheres or microparticles and implantable films or sticks without the need for application of the method, which causes the formation of multiphase emulsions or non-aqueous two-phase systems. Preferably the microparticles are produced by (a) dissolving the composition in an aprotic, miscible with water, an organic solvent; (b) mixing the organic solvent in water, and (C) the selection of particles from the water. In preferred embodiments, the implementation of an organic solvent selected from the group consisting of acetone, acetonitrile, tetrahydrofuran, dimethylformamide and dimethoxyacetophenone.

In other preferred embodiments, the implementation of the ionic molecular conjugate complex polyester/polypeptide capable of visualaid the re, 20 days and more preferably for up to 95 days, but not less than 7 days. In some other preferred embodiments, the implementation of release of therapeutic ionic molecular conjugate is essentially monophasic.

In this invention it is preferable that the composition of continuous release get through (a) obtaining a complex polyester having a free COOH group, and bioactive polypeptide containing at least one effective ionogenic amine, and (b) ion complex conjugation of polyester with the polypeptide with the formation of ionic molecular conjugate, in which at least 85 wt.% polypeptide present in the composition, is ionno conjugated with complex polyester. Complex polyester may be a polyester, which primarily contains a sufficient number of free COOH groups, or, if there is an insufficient number of such groups for the desired level of peptide accession, complex polyester may (1) to react with, for example, malic, citric or tartaric acid by esterification or functional currency or (2) have at the end of the acid residues, for example, glutaric is. Finally, ionic molecular conjugate complex polyester/polypeptide can be transformed into an implantable film or sticks, or injectable microspheres or microparticles capable of release in vivo polypeptide.

Preferably complex polyester synthesized in the catalyzed or autocatalytically direct condensation of one or more hydroxyacids such as glycolic acid and lactic acid, in the presence of a predetermined concentration of the polycarboxylic hydroxy acids, such as malic acid, citric acid or tartaric acid. Educated thus polyesters containing hydroxyl end groups with attached acid residues and hydroxyl groups preferably have a partially or fully-terminal acidic residues.

Polyesters can also be synthesized in catalisano polymerization of lactones with a ring opening or in the polymerization of cyclic monomers, such as-caprolactone, p-dioxanone, trimethylantimony, 1,5-dioxan-2-he or 1,4-dioxan-2-he, in the presence of the initiator of the chain, for example, polycarboxylic hydroxy acids.

Another method of synthesis consists in the accordance polycarboxylic acid.

Another method of synthesis involves the reaction of organic polycarboxylic acid with a pre-formed complex polyester.

In the above preferred embodiments, the implementation of complex polyester with attached at the ends of acidic residues is the ratio of end groups, carboxyl to hydroxyl, more units and approaching infinity (i.e. removing all hydroxyl groups) with an average degree of polymerization between 10 and 300 and, in particularly preferred embodiments, implementation, between 20 and 50.

Alternative complex polyester is capable of forming ionic molecular conjugate with a bioactive polypeptide in the processing base, for example NaOH.

Preferably ionic molecular conjugate complex polyester/polypeptide synthesized by direct interaction between complex polyester, for example, in the free form and the polypeptide, for example, in free form in a suitable liquid medium. In other preferred embodiments of the invention suitable solvents for the formation of the conjugate can be a mixture of aprotic solvent (e.g. acetone, tetrahydrofuran (THF) or dimethyl ether etilenglikolevye. Preferably the polypeptide is a salt of monocarboxylic acid having a pKa greater than or equal to 3.5. Preferably, the polypeptide contains at least one effective ionogenic amine group.

In preferred embodiments, the implementation of the polypeptide is from 1 to 50 wt.% and preferably 10 to 20 wt.% ionic molecular conjugate complex polyester/polypeptide. In the preferred directions of the available carboxyl groups of a complex of the polyester partially neutralized with alkali metal ions or organic bases. In other preferred embodiments, the implementation of the alkaline processing provides the dissociation chain complex of the polyester and the formation of low-molecular binding sites.

In another aspect this invention relates to a complex of the polyester (designated as complex polyester (A) containing one or more free COOH groups and characterized by the ratio of carboxyl and hydroxyl unit, and named the polyester contains a member selected from the group consisting of L-lactic acid, D-lactic acid, DL-lactic acid, malic acid, citric acid,-caprolactone, p-dioxanone,-kapronovaya, substituted or unsubstituted trimethylhexanoate, 1,5-dioxan-2-it, 1,4-dioxan-2-it, glycolide, glycolic acid, L-lactide, D-lactide, DL-lactide, mesolectal and any optically active isomers, racemates or copolymers provided that citric acid-caprolacton and glycolide are elements of a complex of the polyester. The preferred option above complex of the polyester (designated polyester) is a complex polyester, which contains citric acid,-caprolacton and glycolide. The preferred option directly above complex of the polyester (designated polyester (C) is that the ratio of-caprolactone and glycolide in the polyester ranges from 90 to-caprolactone : 10 glycolide to 99-caprolactone : 1 glycolide. Preferred complex polyester (designated polyester D) is one where the ratio of-caprolactone and glycolide in the polyester is 97-caprolactone : 3 glycolide.

In another aspect this invention is directed to a composition comprising the polyester A, polyester B, p is non in itself, at least one effective ionogenic amine, in which at least 50 wt.% polypeptide present in the composition, are ion conjugated with polyester.

The preferred option directly above compositions is that biologically active polypeptide selected from the group consisting of LHRH, somatostatin, bombezin/fiberglass, calcitonin, bradykinin, Galanina, MSH, GRF, Amylin, tachykinins, secretin, PTH, CGRP, neuromedin, Rtgr, glucagon, neurotensin, ACTH, GHRP, GLP, VIP, RASER enkefalina, PYY, motilin, substance P, NPY, TSH and their analogues or fragments.

The preferred implementation described directly above compositions is that biologically active polypeptide selected from the group consisting of LHRH, somatostatin and its analogs or fragments.

The preferred option directly above compositions is that the LHRH analogue is a peptide of the formula pGlu-His-Trp-Ser-Tyr-D-Trp-Leu-Arg-Pro-Gly-NH2and the analogue of somatostatin is a peptide of the formula H2N-D-Nal-Cys-Tyr-Trp-Lys-Val-Cys-Thr-NH2in which two balance s analogue of somatostatin related to each other.

Predpochetaet in the form of sticks.

The preferred option directly above compositions is that the wand has a coating of complex polyester.

The preferred option directly above compositions is that the polyester coating sticks is an absorbable complex polyester.

The preferred option directly above composition is that of absorbable complex polyester contains one or more free COOH groups and is characterized by the ratio of carboxyl and hydroxyl larger units, in which the above-mentioned complex polyester contains a member selected from the group consisting of L-lactic acid, D-lactic acid, DL-lactic acid, malic acid, citric acid, tartaric acid,-caprolactone, p-dioxanone,-Caproic acid, oxalate alkylene, oxalate cycloalkene, succinate of alkylene,-hydroxybutyrate, substituted or unsubstituted trimethylhexanoate, 1,3-dioxan-2-it, 1,4-dioxan-2-it, glycolide, glycolic acid, L-lactide, D-lactide, DL-lactide, mesolectal and any optical aktivnyiy composition is what absorbable polyester coating sticks is the same as a complex polyester included in the composition.

In another aspect this invention is directed to a complex polyester (designated polyester (E) containing one or more free COOH groups and characterized by the ratio of carboxyl and hydroxyl unit, and named the complex polyester contains a member selected from the group consisting of L-lactic acid, D-lactic acid, DL-lactic acid, malic acid, citric acid, tartaric acid,-caprolactone, p-dioxanone,-Caproic acid, oxalate alkylene, oxalate cycloalkene, succinate of alkylene,-hydroxybutyrate, substituted or unsubstituted trimethylhexanoate, 1,5-dioxan-2-it, 1,4-dioxan-2-it, glycolide, glycolic acid, L-lactide, D-lactide, DL-lactide, mesolectal and any optically active isomers, racemates or copolymers provided that tartaric acid is part of a complex of the polyester. A preferred variant of the above-described complex polyester (designated polyester F) is that the polyester contains L-lactic acid iolevel acid. Another preferred variant implementation of complex polyester E (designated as polyester (G) lies in the fact that the polyester contains tartaric acid,-caprolacton and trimethylantimony. The preferred option directly above polyester (designated polyester N) lies in the fact that the ratio of-caprolactone and trimethylantimony in the polyester ranges from 90 to-caprolactone : 10 trimethylhexanoate to 99-caprolactone : 1 trimethylhexanoate. The preferred option directly above complex of the polyester (designated polyester (I) is that the ratio of-caprolactone and trimethylantimony in the polyester is 98-caprolactone : 2 trimethylhexanoate.

In another aspect this invention is directed to a composition comprising a polyester, the polyester F, G polyester, polyester N or polyester I, ion conjugated with one or more bioactive polypeptides containing at least one effective ionogenic amine, in which at least 50 wt.% polypeptide, prisutstvuyuschee the above composition is that bioactive polypeptide selected from the group consisting of LHRH, somatostatin, bombezin/fiberglass, calcitonin, bradykinin, Galanina, MSH, GRF, Amylin, tachykinins, secretin, PTH, CGRP, neuromedin, Rtgr, glucagon, neurotensin, ACTH, GHRP, GLP, VIP, RASER enkefalina, PYY, motilin, substance P, NPY, TSH and their analogues or fragments.

The preferred option directly above compositions is that the bioactive polypeptide selected from the group consisting of LHRH, somatostatin and its analogs or fragments.

The preferred option directly above compositions is that the LHRH analogue is a peptide of the formula pGlu-His-Trp-Ser-Tyr-D-Trp-Leu-Arg-Pro-Gly-NH2and the analogue of somatostatin is a peptide of the formula H2N-D-Nal-Cys-Tyr-Trp-Lys-Val-Cys-Thr-NH2in which the two Cys residue analogue of somatostatin related to each other.

The preferred option directly above composition is that the composition is in the form of sticks.

The preferred option directly above compositions is that the wand has a coating of complex polyester.

Predpochtitelnye polyester contains one or more free COOH groups and is characterized by the ratio of carboxyl and hydroxyl unit, and named the complex polyester contains a member selected from the group consisting of L-lactic acid, D-lactic acid, DL-lactic acid, malic acid, citric acid, tartaric acid,-caprolactone, p-dioxanone,-Caproic acid, oxalate alkylene, oxalate cycloalkene, succinate of alkylene,-hydroxybutyrate, substituted or unsubstituted trimethylhexanoate, 1,5-dioxan-2-it, 1,4-dioxan-2-it, glycolide, glycolic acid, L-lactide, D-lactide, DL-lactide, mesolectal and any optically active isomers, racemates or copolymers.

A preferred aspect directly above composition is that of absorbable polyester coating sticks represents the same complex polyester included in the composition.

Used the term "polypeptide" refers to a protein, peptide, Oligopeptide or synthetic Oligopeptide.

Used the term "polycarboxylic" refers to compounds having more than one carboxyl group, such as malic acid, citric acid and tartaric acid.

Used the term "average degree of polymerization" refers to Kolichestvennyi contains, at least one amine group capable of forming an ion in common conditions.

The term "contains terminal acidic residues" refers to compounds having at the end of the acid.

The term "partially contains terminal acidic residues" refers to compounds in which 1-99% of the hydroxyl end groups are attached acid residues.

The term "fully contains terminal acidic residues" refers to compounds in which 99.9% of their hydroxyl end groups are attached acid residues.

The term "hydroxy acid" refers to any compound containing hydroxyl and carboxyl groups.

The term of monocarboxylic hydroxycitrate" refers to an organic acid one carboxyl group and one or more hydroxyl groups.

The term "polycarboxylic of hydroxycitrate" refers to hydroxycitrate having more than one carboxyl group.

The term "organic azeotropic-distillation propellant" refers to organic liquids that are distilled together with water.

The term "bioactive" refers to a molecule that causes a biological phenomenon or affects the biological event.

The term "acilitate" refers to the formation of a complex of the polyester by condensation of two or more molecules.

Used in the description of the term "absorbable" complex polyester refers to a water-insoluble complex of the polyester, which undergoes dissociation chain in the biological environment with the formation of water-soluble by-products.

This invention relates to new pharmaceutical compositions which are chemically linked biocompatible, biodegradable complex polyester with oligopeptides, polypeptides, peptides and/or proteins in the form of a homogeneous ion product. As a result of chemical bonding of polyester resins with different molecular weights with therapeutic agents chemical characteristics of the composition can be more accurately selected in order to satisfy the requirements of the controlled monophasic release of biologically active polypeptide molecules in vivo. In addition, compositions of the invention are easily optimized for the acquisition of functional properties for better utilization of therapeutically active polypeptide.

Other features and advantages of the invention will become apparent from the following detailed description of preferred embodiments and from the claims.

Brief description of drawings

Fig.1 is an illustration of what the notes.

Fig.2 is an image ionic molecular conjugate describing chemical interaction between the copolymer of lactide/glycolide (Apple type) and Somatuline (BIM 23014).

Fig.3 is a graphical representation of the percentage of peptide released from the ionic molecular conjugates in PBS buffer at 37With over a 28-day period.

Description of the preferred embodiments of the invention

Synthesis

Biodegradable or absorbable polyesters, specially prepared to create the desired chemical reactivity, to provide controlled gidrolizuet chains and extreme ability to contact the oligopeptides, polypeptides or proteins having an overall positive charge at physiological pH, in the proper selection of the constituent monomers, comonomers or somerow with the formation of chains with pre-defined structures and molecular masses.

To obtain the compositions of this invention was applied to the synthesis scheme, consisting of three parts, which correspond to the competence of specialists in this field. Stages include (1) synthesis of polyesters with terminal residues polycarbonate polyethers with terminal residues polycarboxylic acid (or a complex of the polyester, treated base) and biologically active polypeptides and (3) the transformation of the ionic conjugates in implants, rods, microspheres or microparticles capable of release in vivo therapeutic agent within 7 days.

(1) Synthesis of polyesters with terminal residues polycarboxylic acids

Chain polyesters with terminal residues polycarboxylic acids according to the invention was synthesized according to the methods, such as direct condensation of 2-hydroxy acids and polycarboxylic organic acids, stepwise polymerization alkotmany products, polymerization with ring opening of the lactone or mixture of lactones or functional currency organic polycarboxylic acid with a pre-obtained high-molecular complex polyesters (see Fig.1). Describe the synthesis of polyesters with terminal residues polycarboxylic acid according to the aforementioned methods follow.

Direct condensation of 2-hydroxy acids in optically active and/or inactive form and a predetermined amount of polycarboxylic organic acid in the presence and in the absence of inorganic or ORGANOMETALLIC catalyst, for example the condensation of glycolic acid, DL-lactic cibalae monocarboxylic hydroxy acids in the presence of fraction polycarboxylic hydroxy acid in a glass reactor, equipped to ensure a current of dry nitrogen and the mass mixing (designated polyester IA type, see table I). Usually, the polycondensation was carried out at 150-170C for 4 to 72 hours. Stirring of the reaction mixture can be provided with a magnetic stirrer or ozonation of gaseous nitrogen through the polyester mass. The polymerization was continued until, until you have achieved the desired average molecular weight (determined depending on the viscosity of the solution) and/or the number of acidic residues (determined by titration end groups). Analysis of polyester titration end groups was carried out as follows. Examples of polyesters (300-500 mg) was carefully weighed and dissolved in a minimal amount (10-30 ml) of acetone. After dissolving, the solution was diluted to 100 ml benzyl alcohol (Mallinnckrodt, Analytical Reagent) and was titrated until slightly pink in the endpoint (phenolphthalein), using potassium hydroxide in a solution of benzyl alcohol (established normality against the standard model HC1). To determine the number of acid residues, the amount of the basic solution used as the sample (Vs), compared with the volume of the base, ipolito the end of the polymerization complex polyester was isolated and was extracted with water or a dilute aqueous solution of sodium hydroxide from the corresponding organic solution, to remove soluble in water or solubilization low molecular weight chain.

Analysis of polyester using GPC (GPC) was carried out as follows. Average molecular weight (MW; M, m) of the polyesters was determined by GPC using a pump, which supplies the solvent, the model 6000 Waters, and the detector model UV-D Dynamax (Rainin). The experiments were carried out in tetrahydrofuran (Burdick & Jackson, UV grade), using a column of 50 cm10 mm, Jordi Gel DVB 1000(Jordi Associates) at a speed of 1.2 ml/min at 25C. Determination of the peak was carried out at 220 nm and 1.0 AUFS. The column was calibrated using reference standards of narrow zones of polystyrene (Polysciences Inc.) with M m=4000; 9200 and 25000.

Modification of the method of direct condensation entails the application of organic azeotropic-distillation of the displacer and cation exchange resin as a condensation catalyst (designated polyester IB type, see table I). In this way the necessary stage filtration and evaporation to remove the catalyst and azeotropic-distillation separator, respectively. Typical examples of the polyesters obtained according to the described methods, and the corresponding results of the analysis are presented in table I.

When polymerization with ring opening of the lactone or mixture of lactones in the presence of a pre-defined concentration of the polycarboxylic hydroxy acid as the initiator of the chain and catalytic amounts of ORGANOMETALLIC catalyst, for example a mixture of L-lactide, glycolide and DL-malic acid in the presence of octoate tin, used dry cyclic monomers or mixture of cyclic monomers, polycarboxylic gidrokshikislotu and tracking the number of octoate tin (used in the form of a 0.33 M solution in toluene), which in the dry atmosphere without oxygen was transferred into a glass reactor equipped for whom and under nitrogen atmosphere, adhering to the appropriate schema heat, until was not achieved the desired molecular weight (which was measured depending on the viscosity of the solution). On completion of the scheme of the polymerization, the temperature was lowered and the unreacted monomer was distilled under reduced pressure. Then the polyester mass was cooled and removed water-soluble low molecular weight fractions using low-temperature extraction of a suitable organic solution. Then the solution was dried and the solvent was removed. Then he determined the molecular weight depending on the viscosity and the number of acidic residues was determined by titration of end groups. Examples of the polyesters obtained according to this method, and the corresponding results of the analysis are presented in table III.

Functional currency polycarboxylic or polybasic organic hydroxyacids with previously obtained high-molecular esters with a ratio of COOH/IT really appropriate to zero, preferably in the presence of an ORGANOMETALLIC catalyst, such as reaction-melting copolymer 85/15 lactide/glycolide with molecular weight of more than 5000 and COOH/HE1, entails heating a high molecular weight polyester with a predefined amount of polycarboxylic acid or polycarboxylic hydroxy acid in the presence of trace amounts of ORGANOMETALLIC catalyst, such as octoate tin. The reagents were heated above 150C in an atmosphere of dry nitrogen with vigorous stirring until then, until he had completed the functional currency (which was assessed by the depletion of residual unreacted polycarboxylic acid). In fact, the end of the reaction was determined by monitoring the molecular weight (depending on solution viscosity using a capillary viscometer at 28(C) the obtained low molecular weight polyester and the presence of unreacted polycarboxylic acid. The control was carried out by aqueous extraction of a sample of polyester and analysis of the extract using high-performance liquid chromatography (HPLC; HPLC). The levels of the remaining monomer, dimer and polycarboxylic acid was determined by HPLC using a pump solvent model 6000 Waters, and the detector model UV-D Dynamax (Rainin) (205 nm of 1.0 AUFS). Experiments were performed using a buffer with 0.0025 N Na2PO4, pH=3,the Desired complex polyester were isolated and purified, as described above for the polymerization with ring opening. Examples of the polyesters obtained according to this method, and the corresponding results of the analysis are presented in table IV.

Other monomers suitable for the synthesis of polyesters used in the invention are L-lactic acid, DL-lactic acid,-caprolactone, p-dioxanone,-hexanoic acid, trimethylantimony, 1,5-dioxan-2-it, 1,4-dioxan-2-it, glycolic and mesolectal. Examples of initiators used chain and/or modifiers chain include malic acid, citric acid and tartaric acid.

(2) Synthesis of ionic complex conjugate polyester/polypeptide by ionic interactions polyesters with terminal residues of polycarboxylic acids and biologically active polypeptides

Used the above-described biodegradable polyesters with terminal residues of polycarboxylic acid to produce ionic molecular conjugates with mono - or polycarboxylate the oligopeptides, polypeptides or proteins with available effective ionogenic amine groups (see Fig.2). In addition, any complex polyester was IEM, such as 0.1 N NaOH. Such processing is disclosed acid groups of the polyester for ionic interaction with a cationic polypeptide in many areas.

Thus, the formation of these conjugates was achieved as a result of direct molecular interaction between the components in a suitable solvent with or without pre-treatment of polyester inorganic base, in order to maximize its ability to bind an essential medicine. As noted above, the ionic interaction of the ionic components of the conjugate increases within the difference in the values of their pKa.

The polyester was dissolved in a suitable aprotic solvent in the concentration range from 2 to 20% (wt./about.) Such a solvent should dissolve polyesters, but also partly be mixed with water. Suitable solvents used for this purpose include tetrahydrofuran, acetone, and dimethyl ether of ethylene glycol. To the resulting solution was added an aqueous solution of base such as the hydroxide or carbonate of sodium, potassium or ammonium, in order to maximize the binding capacity of polyester. In General, the amount of added base corresponded to the number of acid represented by the level proefi-base was added an aqueous solution of the peptide or salt of the peptide at levels filling the peptide/polyester 2 to 50 % (wt./wt.) (peptide/polyester). The resulting mixture was stirred for a period of time (up to 3 hours), and then the solvent was removed and the product dried under vacuum. The resulting material can then be further processed for metered dose of the composition. Suppose that the resulting pharmaceutical composition must be chemically standard compositions entirely of ionic molecular conjugates and essentially be free from microscopically or macroscopically dispersed domains of the active drug in a biodegradable matrix. Examples of the obtained ionic molecular conjugates and the corresponding results of the analysis are presented in table V.

(3) the transformation of the ionic conjugates in implants, rods, microspheres or microparticles capable of release in vivo therapeutic agent, at least within 20 days from monophasic profile

Salt ionic conjugates of the invention can be converted into (A) sterile injectable microspheres (in the presence of from 0.1 to 10% solid polyhydride alcohol as an adjunct treatment or without it), containing from 1 to 50 wt.% polypeptide, which may be released on) is 2 weeks. (C) sterile implantable film obtained by casting, molding or extrusion with or without pharmacologically inert auxiliary processing tools and is capable of providing a release profile similar to the profile described in (A) and (C) sterile injectable sticks obtained by extrusion or pressing, capable of providing a release profile similar to the profile described in (A). Besides sticks can be covered with a complex polyester, to provide an additional layer of control over the rate of release of therapeutic agent. Preferably sticks covered absorbable polyester; more preferably absorbable polyester is the same as defined in the description, and most preferably covering absorbable polyester is the same as the polyester, a prisoner in the wand.

A study release in vitro

The specimens are dried and powdered ion conjugate, weighing 50 mg each, were placed in scintillation vials with a diameter of 25 mm Aliquots in 5 ml of modified PBS buffer (PBS buffer: 2,87 g Na2HPO4, 0,654 g Pan2RHO4, 5.9 g of NaCl, 0.5 g NaN3sufficient amount of deionized water on the tion at 120 rpm and 37C. the Vials were periodically removed, poured the fluid and refilled with fresh PBS solution. The number of released peptide was determined from decantering solutions PBS by HPLC.

Extraction of peptides from the ion conjugates

A sample of 50 mg of ionic molecular conjugate was mixed in 20 ml of methylene chloride. The mixture consistently was extracted with portions of 50 ml, 20 ml and 20 ml of 2 N acetic acid. The acetic acid extracts were combined and analyzed in respect to the content of peptides by high-performance liquid chromatography (HPLC; HPLC). Peptide analysis HPLC was carried out as follows. The HPLC analysis was carried out using the pump of the solvent, the model M-45 Waters, and the detector EAT MACS Science 700 at a wavelength of 220 nm and 1.0 AUFS. The peptides investigated using Lichrospher (EM separations) C18, 100A, 5 μm, column 25 cm4.6 mm and 30% acetonitrile/0.1% of TFA as isocrates eluting buffer.

Below are the results (table VI) in vitro studies demonstrating the amount of peptide released during the 28-day period for ionic molecular conjugates 49:49:2 L-lactic/glycolic/malic/D-Thr6[LHRH] (example 8), 49:49:2 L-lactic/glycolic/malic/somatostatin - analogue, which inhibits.gif">

The number of peptides in ion conjugates

Ion-related peptides in the products of the conjugate was measured by dissolving 10 mg of the sample 5.7 ml of a mixture (9:1) of acetone and 0.1 M aqueous triperoxonane acid. The solutions were mixed by rotation at approximately 25With approximately 15-24 hours, and then filtered through a 5 μm sleeve Teflon filters. Then the filters were analyzed on the content of the peptide high-performance liquid chromatography (HPLC). Peptide analysis by HPLC was performed using Millipore, model 777 Wisp Autosampler, pump, model 510, and a set of UV detectors, model 486, at 220 nm. Peptides were investigated on Lichrospher (EM Separations) column (25 cm4, 6 mm, C18, 5 μm, 100the flow velocity of 1.0 ml per minute, using a 35% acetonitrile in buffer 0.14% sodium perchlorate as isocrates eluting system. The peptides were evaluated quantitatively by comparing the peak area of precision in the sample area entered peptide standard.

Use

Any, in the description of the ionic conjugates acid polyesters/polypeptide can be administered to the recipient only alone or in combination with pharmaceutically p is ositories or nasal, therapeutic drug is administered in accordance with condition that should be treated. The concentration of the composition in the preparations of the invention varies depending on a number of factors, including the dose, which should be introduced, and the way of introduction.

I believe that the experts in this field, using the preceding description, no further development will be able to apply this invention in full. Therefore, the following implementation options should be construed as illustrating and not limiting description.

Example 1. The direct condensation - synthesis of 50/50 poly(D,L-lactic-co-glycolic acid) catalyzed by amberlyst 15.

D,L-Lactic acid (85% aqueous mixture; 13,7 g, 0.13 mol) was mixed with glycolic acid (10 g, 0.13 mol) in a round bottom flask, equipped with magnetic stirrer, water trap Dean-stark (Dean-Stark) and a condenser cooled by water. Was added toluene (100 ml) and granules of amberlist 15 (100 mg) and the mixture is boiled under reflux in nitrogen atmosphere for 72 hours, removing water from the mixture. The mixture was cooled, toluene decantation of the solidified mass and the product was dissolved in methylene chloride (250 ml). The methylene chloride was treated with activated charcoal (Darco, 500 mg), filtered and �https://img.russianpatents.com/chr/176.gif">With to obtain a white powder (harin l3=0,3, acid #=2439, TD=12C).

Example 2. The direct condensation synthesis 49/49/2 poly(L-lactic-co-glycolic/citric), catalyzed by amberlyst 15.

Using the system, similar to the above, L-lactic acid (88% aqueous mixture; 25,6 g, 0.25 mol) was mixed with glycolic acid (19.2 g, 0.25 mol), citric acid monohydrate (2,33 g to 0.011 mol), granules of amberlist 15 (500 mg) and toluene (150 ml) in a round bottom flask. The mixture was heated under stirring to the boiling temperature under reflux for 51 hours, removing water by using traps Dean-stark. Toluene decantation of the semi-solid product. Complex polyester was dissolved in acetone (300 ml) and filtered and dried on a rotary evaporator. Solid polyester then re-dissolved in methylene chloride and washed twice with water (2150 ml) to remove soluble oligomers. The organic solution was concentrated on a rotary evaporator and the product was dried in vacuum to obtain a white solid (see table I, the polyester type IB, polymer #4) (harin l3=0,11, acid #=842, TD=15Using a cylindrical ampoule, with a capacity of 150 ml with fitting air impinger, L-lactide (20 g, 0,139 mol) was mixed with glycolic acid (7,1 g, 0,093 mol) and (d,l)-malic acid (1.0 g, 0,0075 mol). The mixture was stirred by bubbling nitrogen through the inlet of impinger (100 ml/min) and heated from 25 to 155With over 100 minutes. The reaction temperature was maintained at 155With over 70 hours and the water of the polymerization was removed to a cold trap on the exhaust manifold reactor. After 70 hours, the reaction mixture was cooled to 100C and poured into a chilled receiver stainless steel for hardening. Solid polyester was then dissolved in methylene chloride and washed twice with water (2150 ml) to remove soluble oligomers. The organic solution was concentrated on a rotary evaporator and the product was dried in vacuum to obtain a white solid (see table II, polyester type II polymer #2) (harin l3=0,13, acid #=1800, Th=27C).

Example 4. The method of polymerization with ring opening - synthesis of 75/25 poly(L-lactide the slot (0,3042 g, 0,00227 mol) and the catalyst octoate tin (0.33 M in toluene, 67 μl, of 0.022 mmol) in dry nitrogen atmosphere was introduced in a glass vial with a magnetic stir bar. The system was purged with N2and was pumped vacuum several times before closing the ampoule. The reagents were then melted at 140C and the melt was heated at 180, 190, 180, and 150With over 1; 4,5; 12 and 2 hours, respectively. After cooling to room temperature, the polyester was re-heated to 110With in a vacuum at a pressure less than 1 mm RT.article within approximately one hour to remove monomer, re-cooled at room temperature, put in liquid nitrogen, were isolated and dried under vacuum (harin l3=0,20, acid #=2560, TD=39C).

Example 5. The method of polymerization with ring opening - synthesis of 50/50 poly(D,L-lactide-co-glycolide), initiated by citric acid.

D,L-Lactide (10.0 g, 0,0694 mol) was mixed with glycolide (8.06 g, 0,0694 mol), citric acid (1.07 g, 0,00555 mol) and catalyst (octoate tin (0.33 M in toluene, 84 μl, 0,0278 mmol) in dry nitrogen atmosphere in a glass vial with magnetic stirrer and closed under vacuum. The reagents were melted and the room temperature, extinguished in liquid nitrogen, were isolated and dried (harin l3=0,26, acid #=970, TD=23C).

Example 6. The method of polymerization with ring opening - synthesis of 50/50 poly(D,L-lactide-co-glycolide), initiated 1,6-hexandiol.

Using the system, similar to the above system, D,L-lactide (10.0 g, 0,0694 mol), glycolide (8.06 g, 0,0694 mol), 1,6-hexanediol (0,656 g, 0,00555 mol) and octoate tin (0.33 M in toluene, 84 μl, 0,0278 mmol) in dry nitrogen atmosphere was introduced in a glass ampoule, which was subsequently closed in a vacuum. The reaction mixture was heated at 150, 185, 150, 120C for 0,5; 4; 1; 5 and 3 hours, respectively. The obtained polyester was recovered and dried (see table III, polyester type III, polymer #5) (harin l3=0,39, acid #=10138, TD=30C).

Example 7. The method of functional currency is the synthesis of carboxyl-containing 50/50 poly(D,L-lactide-co-glycolide).

50/50 poly(D,L-lactide-co-glycolide) (Boehringer A001, 8 g), citric acid (0.8 g, 4,16 mmol) and octoate tin (2 drops) were added in a glass ampoule in an atmosphere of dry nitrogen and closed. The mixture was heated at 150C for 4 hours, cooled to room temperature/img>harin l3=0,26, acid #=670, Tg=23C).

Example 8. Synthesis 49:49:2 L-lactic/glycolic/malic (see table I, polymer#4) and D-Trp6[LHRH]ionic molecular conjugate.

500 mg 49:49:2 L-lactic/glycolic/malic acid (synthesized by direct condensation; M m=9500; acid #=1420) was dissolved in 10 ml of acetone (Mallinckrodt Analytic Reagent). Added a portion of a 0.1 N solution of sodium hydroxide (1,14 ml) and the mixture was stirred at room temperature for 15 minutes. A solution of 100 mg of D-Trp6[LHRH] (BIM-21003 peptide I; the content of the Foundation of 87%, the acetone 7%) in 1.0 ml of water was added and the mixture was stirred for 1 hour at room temperature. Then the solvents were removed first by using Rotovap (Rotovap) when T<40And then in a desiccator for 1 hour at room temperature in a vacuum of 1 mm RT.article The dried solid was ground and stirred in 100 ml of deionized water and was isolated by filtration. The aqueous filtrate was analyzed by HPLC and found that it contains <1 mg of soluble peptide. The solid material was dried for several days under vacuum to obtain 540 mg of a white powder. The powder used in in vitro studies (see table VI, example 8).

An example of a statin/analog inhibiting tumor.

100 mg 49:49:2 L-lactic/glycolic/malic (synthesized by direct condensation; M m=9500; acid #=1420) was dissolved in 2 ml acetone (Mallinckrodt Analytic Reagent). Was added portion of 0.1 N solution of sodium hydroxide (0,32 ml) and the mixture was stirred at room temperature for 15 minutes. Solution was added 20 mg of somatostatin/analogue, which inhibits tumor (BIM-23014 peptide II; the content of the Foundation of 83%, the acetone 9,8%), 1.2 ml of water and the mixture was stirred for 1 hour at room temperature. Then solvents were removed first by using rotovap at T<40And then in a desiccator for 1 hour at room temperature in a vacuum of 1 mm RT.article The dried solid was ground and stirred in 20 ml of deionized water and was isolated by filtration. The aqueous filtrate was analyzed by HPLC and found that it contains <0.05 mg of soluble peptide. The solid material was dried for several days under vacuum to obtain 106 mg of a white powder. The powder was grinded and used in the study release in vitro (see table VI, example 9).

Example 10. Synthesis of 73.5:24,5:2 poly-L-lactide/glycolic/malic (see table II, polymer #2) and ionic molecular conjugate D-Trp6[LHRH].6[LHRH] (BIM-21003; the content of the Foundation of 87%, the acetone 7%) in 2 ml of water and the mixture was stirred for 90 minutes. The solvent was removed and the resulting solid is triturated in deionized water as in example 8, found that there is less than 1% salt soluble peptide. Selected solids were dried for 4 days in vacuum to obtain 839 mg of white powder. The powder was grinded and used for research release in vitro (see table VI, example 10).

Example 11. The formation of microparticles 1,50 ion conjugate the peptide-polymer polyester L-lactide/glycolide/D,L-lactic acid (65:33:2).

The conjugates synthesized by polymerization with ring opening, as described in example 4 (M m=4700, the degree of polydispersity = 1.3, which was determined by GPC 501 cm column with a linear mixed layer Jordi Gel, eluent THF, the detector light scattering Wyatt Dawn Mini dn/dc=0,05, acid #1475 - titration, TD=42C), and dissolved in 40 ml of acetone. Acid groups are neutralized 2.0 ml, 0.5 M solution g is and 11.5%) in 20 ml of water Milli-Q under stirring was slowly added to the polymer solution. To prevent deposition while adding peptide was also added additional portions 40 ml of acetone. Clear colorless solution was stirred for one hour and then evaporated in a vacuum until dry. The obtained white solid was re-dissolved in a mixture of 20 ml of acetone and 2 ml of water Milli-Q for the formation of a transparent solution. The resulting solution was injected through a 0.2 µl Teflon filter in a fast peremeshivayte tank with 500 ml of Milli-Q water at 4C. the Phase of the complex polymer/peptide were immediately divided into small particles upon contact with water. After stirring the suspension for 30 minutes at 4With the remaining acetone was removed under reduced pressure and the solids were isolated by centrifugation, re-suspended with 100 ml of Milli-Q water and re-centrifuged. Selected solids were dried by lyophilization to obtain 1530 mg white free flowing powders. The average particle size of 2-100 μm. It is shown that TD ionic conjugate is located at 53C. According to HPLC determined that the total residual (unbound) peptide in all water supernatant is 63 mg How it was determined what isua extraction acetone/0.1 M TFA, found that the percentage extracted from the peptide conjugate is 16.9 wt.%. Thus, the resulting conjugate retains at 84.8% of the ion (extracted) the nature of the relationship.

The delivery system in the form of a wand type 1 (CONC2 and CGC1)

Example A-1. Getting copolymer 97/3 caprolacton/glycolide (CGC1), initiated by citric acid.

A round bottom flask, equipped with a mechanical stirrer, double-flame dried and purged with dry argon. The flask was uploaded-caprolactone (1,455 mol, 166 g), glycolide (0,08865 mol, 10.3 g), citric acid (0,075 mol, of 14.4 g) and octoate tin (0,0003 mol, 375 μl of 0.8 M solution in toluene). The polymerization was performed using the following scheme: in an argon atmosphere loaded material was heated from room temperature to approximately 150C for 1 hour and 20 minutes after fusion with continuous stirring (70 rpm). The load was maintained at approximately 150With over 11.5 hours. After completion of the polymerization, a small amount of unreacted monomer was distilled at approximately 120C for 15 minutes in vacuum (0.1 mm, RT.cent.). Matera is n=52C) and titration of carboxyl groups (average equivalent weight =623 Yes).

20 g of polymer was dissolved in a 50.0 ml of acetone and the solution was besieged with stirring in a bath with ice. The solid product isolated by filtration.

The purified product was analyzed using GPC (MP=4214, M m=9688), DSC (Tn=45,2C) and titration (average equivalent weight =780).

Example B-1. Obtaining ionic conjugate (CONC1).

1.5 g of the pure polymer (CGC1) was dissolved in 7.5 ml of acetonitrile in a glass vial. In a separate vial 250.0 mg LHRH-acetate was dissolved in 1.5 ml of distilled water. The dissolved polymer was filtered through a 0.45 µm syringe filter Acrodisc in a bottle containing sodium carbonate to neutralize the LHRH-acetate). A solution of LHRH was added dropwise to the solution filtered polymer. The combined solution was stirred with a magnetic rod for approximately 1.5 hours at room temperature. The conjugate was besieged by adding it dropwise in peremeshivayte isopropyl alcohol (IPA), cooled with liquid nitrogen. The precipitate was collected by centrifugation and dried overnight in vacuum. The output of the conjugate was $ 73.5%. The conjugate was investigated by DSC (Tn=50,9Example C-1. Receipt of the delivery system in the form of sticks.

Ionic conjugate (0,3987 g CONC2) and the polymer (1,206 g CGC1) was mixed by gentle grinding and melted together at about 58C in the heating block. The molten material was mixed, and then was sucked into the capillary tube 18G and left for cooling. The material was extruded and cut into segments, which contained the appropriate dose of the drug and placed in sterile spiral needles 10-gauge (prepared for injection). All stages of the example C-1 was performed in a fume hood with laminar flow. The content of LHRH in the sticks was 2.5%.

The delivery system in the form of a wand type 2 (CONC2 and CGC1)

Example A-2. Getting copolymer 97/3 caprolacton/glycolide (CGC1), initiated by citric acid.

This example used the same polymer (CGC1), obtained in example a-1.

Example B-2. Obtaining ionic conjugates (CONC2).

CONC2 was obtained according to the method described in example-1. Elemental analysis showed that the percentage of nitrogen is 2,31%. Based on these data, the contents of LHRH was of 12.76%.

Example C-2. Receipt of the delivery system in the form of sticks.

CONC2 (0,1854 g) and 0,5565 g purified CGC1 mechanically mixed, and then heated p is s 18-gauge and said piston. Sticks cut into segments, which contained the appropriate dose of the drug, and were placed in sterile spiral needle gauge 18 (prepared for injection). All stages of the example C-2 was performed in a fume hood with laminar flow. The content of LHRH in the sticks was 3.2%.

The delivery system in the form of sticks 3 type

Example A-3. Obtaining a copolymer (STT) 98/2 caprolacton/trimethylantimony (TMS), initiated by tartaric acid.

A round bottom flask, equipped with a mechanical stirrer, three times was flame dried and purged with dry argon. The flask was uploaded-caprolactone (1,47 mol, 168 g), TMS (0.03 mol, of 3.06 g), tartaric acid (0,0142 mol, 2,134 g) and octoate tin (0,0003 mol, 375 μl of 0.8 M solution in toluene). The polymerization was performed using the following scheme: by injecting argon loaded material was heated from room temperature to approximately 150With approximately 1 hour of time mixing the molten reaction mixture (60 rpm). The temperature was maintained at approximately 150With over 9 hours. Unreacted monomer was distilled at approximately 100With approximately the emer was analyzed by GPC (MP=13221, M m=35602).

Example B-3. Obtaining ionic conjugates (CONCTT1).

1.5 g of purified polymer from example a-3 was dissolved in 7.5 ml of acetonitrile in a glass vial. In a separate vial 250 mg LHRH-acetate was dissolved in 1.5 ml of distilled water. The dissolved polymer was filtered through a 0.45 µm syringe filter Acrodisc into the vial containing 56,5 mg of sodium carbonate (to neutralize LHRH-acetate). A solution of LHRH was added dropwise in the filtered polymer solution. The combined solution was stirred by a magnet within approximately 3 hours at room temperature. The conjugate was besieged by adding it dropwise in peremeshivayte IPA, cooled with liquid nitrogen.

The precipitate was collected by centrifugation and dried overnight in vacuum.

The output of the conjugate was 81,1%. Elemental analysis of the material showed 2,04% nitrogen. Based on this established that the content of LHRH is 11.3%.

Example C-3. Receipt of the delivery system in the form of sticks.

ST (0,8909 g) was melted at approximately 55C. To the melt was added 0,2250 g CONCTT1 and the whole system was heated to approximately 65C. the Molten system then was sucked into the capillary tube 18-gauge and said piston. Sticks cut the e needle gauge 18 (prepared for injection). All stages of the example C-3 was performed in a laminar flow hood. The content of LHRH in the sticks was 2.3%.

The delivery system in the form of sticks 4 type

Example A-4. Getting copolymer 94/6 caprolacton/glycolide (CGT6), initiated by tartaric acid.

A round bottom flask, equipped with a mechanical stirrer, three times was flame dried and purged with dry argon. The flask was uploaded-caprolactone (1,41 mol, 161 g), glycolide (0.09 mol, 10.4 g), tartaric acid (0,005 mol, 0.73 g) and octoate tin (0,0003 mol, 375 μl of 0.8 M solution in toluene). The polymerization was performed using the following scheme: by injecting argon boot material was heated from room temperature to approximately 150With approximately 1 hour of time mixing the molten reaction mixture (60 rpm). The temperature was maintained at approximately 150C for 1 hour. The temperature was then raised to approximately 180With approximately four hours. The material was cooled to approximately 107With and left in vacuum at 1.5 mm RT.article approximately 1.5 hours. The material was poured into the jar PI=26254, M m=68101).

Example B-4. Obtaining ionic conjugates (CONCTT2).

CONCTT2 was obtained as described in example 1, but using LHRH-acetate and copolymer from example A4.

Example C-4. Receipt of the delivery system in the form of sticks.

CGT6 (1.4 g) and CONCTT2 (0,4779 g) was heated to approximately 57With, cooled, crushed, and then re-heated to the same temperature. Then the fused system was sucked into the capillary tube 18-gauge and said piston. Sticks cut into segments, which contained the appropriate dose of the drug, and were placed in sterile spiral needle gauge 18 (prepared for injection). All stages of the example C-4 was performed in a laminar flow hood. The LHRH content was 2.8%.

Example D-4. Floor wand system-4 using inert precursor copolymer.

CGT6 (1.4 g) was dissolved in 1.5 ml dichloromethane. Sticks from example C-4 was immersed in the obtained polymer solution was immediately removed and dried in a laminar flow fume hood at ambient conditions.

From the above description specialists in this field can easily ascertain the essential characteristics of this invention without deviation from the spirit and competencelevel and conditions. Thus, other ways of implementation are also within the claims.

Claims

1. Complex polyester containing one or more free carboxyl groups and characterized by the ratio of carboxyl and hydroxyl groups of more units constituting the copolymer-caprolactone and glycolide at a ratio (90-99):(1-10) obtained in the presence of initiator citric acid.

2. Complex polyester under item 1, characterized in that the ratio of-caprolactone to glycolide is 97:3, respectively.

3. A composition comprising a complex of the polyester under item 1 or 2, ion conjugated with one or more bioactive polypeptides containing at least one effective ionogenic amine, in which at least 50 wt.% polypeptide present in the composition, are ion conjugated with complex polyester.

4. The composition according to p. 3, wherein the bioactive polypeptide is chosen from the group consisting of LHRH, somatostatin, bombezin/fiberglass, calcitonin, bradykinin, Galanina, MSH, GRF, Amylin, tachykinins, secretin, PTH, CGRP, neuromedin, Rtgr, glucagon, neurotensin, ACTH, GHR fact, that bioactive polypeptide, selected from the group consisting of LHRH, somatostatin and its analogs or fragments.

6. The composition according to p. 5, wherein the LHRH analogue is a peptide of the formula pGlu-His-Trp-Ser-Tyr-D-Trp-Leu-Arg-Pro-Gly-NH2and the analogue of somatostatin is a peptide of the formula H2N-D-Nal-Cys-Tyr-Trp-Lys-Val-Cys-Thr-NH2,in which the two Cys residue analogue of somatostatin related to each other.

7. The composition according to p. 4, characterized in that it is in the form of sticks.

8. The composition according to p. 7, characterized in that the said wand has a coating of complex polyester.

9. The composition according to p. 8, characterized in that the polyester coating sticks is an absorbable complex polyester.

10. The composition according to p. 9, characterized in that the absorbable complex polyester contains one or more free carboxyl groups and is characterized by the ratio of carboxyl to the hydroxyl unit, which is called the complex polyester contains a member selected from the group consisting of D-lactic acid, DL-tartaric acid, citric acid, tartaric acid,-caprolactone, trimethylhexanoate, glycolide, and any optically active isomers, racemates or copolymerisation, as complex polyester included in the composition.

12. The composition containing the ester under item 2, the ion - conjugated with one or more bioactive polypeptides containing at least one effective ionogenic amine, in which at least 50 wt.% polypeptide present in the composition, are ion conjugated with complex polyester.

13. Complex polyester containing one or more free carboxyl groups and characterized by the ratio of carboxyl and hydroxyl groups of more units constituting the copolymer selected from the group comprising the copolymer-caprolactone and trimethylantimony at a ratio of 90:10 to 99:1, the copolymer-caprolactone and glycolide when the ratio of 94:6, obtained in the presence of initiator - tartaric acid.

14. Complex polyester on p. 13, characterized in that the ratio of-caprolactone to trimethylantimony is 98:2, respectively.

15. A composition comprising a complex of the polyester under item 13 or 14, ionic conjugated with one or more bioactive polypeptides containing at least one effective ionogenic amine, in which at least 50 mA the position p. 15, wherein the bioactive polypeptide is chosen from the group consisting of LHRH, somatostatin, bombezin/fiberglass, calcitonin, bradykinin, Galanina, MSH, GRF, Amylin, tachykinins, secretin, PTH, CGRP, neuromedin, Rtgr, glucagon, neurotensin, ACTH, GHRP, GLP, VIP, PACAP, enkefalina, PYY, motilin, substance P, NPY, TSH and their analogues or fragments.

17. The composition according to p. 16, wherein the bioactive polypeptide is chosen from the group consisting of LHRH, somatostatin and its analogs or fragments.

18. The composition according to p. 17, wherein the LHRH analogue is a peptide of the formula pGlu-His-Trp-Ser-Tyr-D-Trp-Leu-Arg-Pro-Gly-NH2and the analogue of somatostatin is a peptide of the formula H2N-D-Nal-Cys-Tyr-Trp-Lys-Val-Cys-Thr-NH2in which the two Cys residue analogue of somatostatin related to each other.

19. The composition according to p. 16, characterized in that it is in the form of sticks.

20. The composition according to p. 19, characterized in that the said wand has a coating of complex polyester.

21. The composition according to p. 20, characterized in that the polyester coating sticks is absorbed by the complex polyester.

22. The composition according to p. 21, characterized in that the absorbable complex polyester contains one or more free carboxyl groups Headerget element, selected from the group consisting of D-lactic acid, DL-tartaric acid, citric acid, tartaric acid,-caprolactone, trimethylhexanoate, glycolide, and any optically active isomers, racemates or copolymers.

23. The composition according to p. 22, characterized in that the absorbable polyester coating sticks is the same polyester as complex polyester included in the composition.

24. A composition comprising a complex of the polyester under item 14, the ion conjugated with one or more bioactive polypeptides containing at least one effective ionogenic amine, in which at least 50 wt.% polypeptide present in the composition, are ion conjugated with complex polyester.

 

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< / BR>
and the second link has the formula

< / BR>
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22 cl, 17 dwg, 12 ex

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EFFECT: valuable medicinal properties of compound.

1 cl, 2 tbl, 1 ex

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4 cl, 2 ex

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36 cl, 4 tbl, 4 ex

FIELD: organic synthesis.

SUBSTANCE: invention provides novel triterpenoid derivatives having general formula:

(I), wherein, as polymer carrier, are used water-soluble copolymers of N-vinylpyrrolidone with α,β-(methyl)acrylic acid alkyl esters and quaternary ammonium salts thereof having general formula: (II), in which formulas A represents triterpenoid residue belonging to series of acids: betulinic {1}, betulonic {2}, glycyrrhetinic {3}, glycyrrhisic {4}, ursolic {5}, ursonic {6}, oleanolic {7}, oleanonic {8}, meristothropic {9}, diketomeristothropic {10}, macedonic {11}, diketomacedonic {12},equinocystic {13}, or mixture of above-indicated carboxyl-containing triterpenoids, where R1 and R2 are hydrogen or methyl; R3 methyl or ethyl; R4 is C6-C16-alkyl; Hal is iodine, bromine, or chlorine atom; k = 65-95 mol %; l = 0.1-34 mol %; n = 0.5-5.4 mol %; molecular weight is equal to (7-100)·103. Polymer derivatives of above-defined triterpenoids are prepared by reaction of terpolymer II, wherein k = 65-95 mol %; l = 0.1-34 mol %; x = 1.0-34.9 mol %; R1-R4, Hal and molecular weight as above. Reaction is carried out in organic solvent at concentration of terpolymer 1 to 30%, concentration of triterpenoid 0.05 to 3.4%, and molar ratio of motif containing quaternary nitrogen to triterpenoid between 1 and 10. Products are isolated by removing solvent.

EFFECT: expanded synthetic possibilities.

FIELD: immunology.

SUBSTANCE: invention relates to conjugates having interferon-γ activity. Claimed conjugate contains at least one non-polypeptide group, covalent bonded to IFNG polypeptide, wherein polypeptide contains amino acid sequence differ from starting IFNG polypeptide sequence by at least one added or deleted amino acid residue containing group to add non-polypeptide group.

EFFECT: therapy of diseases of improved effectiveness.

34 cl, 1 ex

FIELD: medicine, oncology, immunology.

SUBSTANCE: invention relates to humanized antibodies with ErbB2. Invention involves the development of new humanized antibodies raised to tyrosinase receptors of family ErbB2, and to a composition comprising these antibodies. The advantage of invention involves expanding region in using indicated antibodies in cancer treatment wherein receptor of epidermal growth factor, EGFR, is a target of these antibodies.

EFFECT: valuable properties of antibody.

14 cl, 3 tbl, 13 dwg

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