Platinum nanocompounds and methods of applying thereof

FIELD: chemistry.

SUBSTANCE: invention relates to bio-compatible conjugated polymer nanoparticle, dicarbonyl-lipid compound, compound in form of vesicles, micelles or liposomes, containing multitude of nanoparticles, including said dicarbonyl-lipid compound, method of treating cancer or metastases, biocompatible polymer, as well as to conjugate. Biocompatible conjugated polymer nanoparticle includes main chain of copolymer, with at least one polymer monomer containing two side chains, selected from the group, consisting of carboxylic acid, amide and ester, and sad side chains are separated from each other by 1-1- carbon atoms, oxygen atoms or sulphur atoms, or their any combinations. Said nanoparticle further contains multitude of side chains, covalently bound with said main chain, with said side chains being selected from the group, consisting of monosaccharides, dicarboxylic acids, polyethyleneglycol and their combinations; and multitude of platinum compounds, dissociatedly bound with said main chain. Multiple platinum compounds are connected with said main chain via at least one coordination bond between carbonyl oxygen of carbonyl or amide group of main chain and platinum atom of platinum compound. Said platinum compound is selected from Pt(II) compounds, Pt(IV) compounds and any their combinations. Invention is also aimed at dicarbonyl-lipid compounds, in which platinum compound is dissociatedly bound with dicarbonyl compound. Invention is also aimed at method of treating cancer and metastases. Methods include selection of subject, requiring treatment of cancer or metastases, and introduction to subject of effective amount of nanoparticles, compounds or compositions of the invention.

EFFECT: obtaining biocompatible conjugated polymer nanoparticles for chemotherapeutic platinum-based preparation.

40 cl, 1 tbl, 29 dwg, 12 ex

 

This application claims the priority and benefit under 35 U. S. C. 119(e) of U.S. provisional application 61/149,725, filed February 4, 2009 and 61/240,007, filed September 4, 2009, the contents of which in its entirety is introduced into the present description by reference.

The present invention is carried out under the grant "Era of Hope Scholar Award from the U.S. Department of defense number W81XWH-07-1-0482 and Grand Postdoctoral Award from the U.S. Department of defense number W81XWH-09-1-0728. The U.S. government has certain rights to this invention.

The technical field to which the invention relates.

The present invention relates to biocompatible conjugated polymer nanoparticles including a main chain of the copolymer, many of the side chains, covalently linked with the main chain, and a lot of platinum compounds, dissociatio associated with the main chain.

The level of technology

Cancer is the second leading cause of death in the United States with projected 1444180 new cases and 565650 deaths in 2008. Cytotoxic agents used in standard chemotherapy indiscriminately affect any cells that are in the fission process, which causes doselimiting toxic effects. There is an urgent need to develop new strategies, more selective effects of the plans on the tumor.

The use of nanovectors holds the potential for a qualitative change in the chemotherapy of cancer by selective effect on the tumor. A number of polymeric nanovectors are currently in development or currently in clinical trials, and greatly alter the pharmacodynamic and pharmacokinetic profile of the active agent. However, most of these polymeric structures reduce the effectiveness of conjugated active agent, based on the increase of absorption by the tumor to improve therapeutic index.

Cisplatin is one of the main directions in chemotherapy regimens for most types of cancer (Kelland L. The resurgence of platinum-based cancer chemotherapy. Nat Rev Cancer. 2007 Aug; 7(8):573-84). However, its use is limited because of severe nephrotoxicity. Besides nanofactory composition based on cisplatin, which is the first line treatment for many types of cancer, is a problem that requires effort.

Disclosure of inventions

Presents the rational design of polymer structures for chemotherapeutic drugs based on platinum, such as cisplatin and oxaliplatin, which causes the self-Assembly of the nanoparticles. Nanoparticle supports the effectiveness of the active agent and compared with cisplatin or oxaliplatin, or carboplatinum has p is increased antitumor activity with reduced systemic and nephrotoxicity when administered intravenously to mice, bearing the tumor. This increase therapeutic index of cisplatin or oxaliplatin possible thanks to nanotechnology, can be used to nanoplatelets in the clinical treatment of many types of cancer.

The invention is directed to biocompatible polymer conjugate nanoparticles, comprising the main chain of the copolymer, many of the side chains covalently associated with the specified main chain, and a lot of platinum compounds, dissociatio associated with the specified main chain. In General, the platinum compound dissociatio connected with the main circuit through connection via a side chain. In some embodiments, the platinum compound is associated with the side chain, at least one coordination bond.

Another aspect of the invention is directed to biocompatible conjugated polymer nanoparticles including a main chain polimolekuly acid (PMA), such as the main chain of poly(isobutylene-alt-maleic acid) (PIMA). The primary circuit comprises from 25 to 50 monomers. Also included are many of the side chains of PEG covalently associated with the specified main chain. Side chains of PEG have a molecular weight of from 200 to 3000 daltons. The number of PEG side chains is from 50% to 100% inclusive of the number of monomer units of the main polymer chain. Also included are the I many side groups of cisplatin or oxaliplatin, dissociatio associated with the main circuit. The number of side groups of cisplatin is from 25% to 75% of the number of monomer units of the main polymer chain.

Still another aspect of the invention is directed to biocompatible conjugated polymer nanoparticles including a main chain of poly(isobutylene-alt-maleic acid). The primary circuit consists of approximately 40 monomers. Also included are many of the side chains of PEG covalently associated with the main circuit. Side chains of PEG have a molecular weight of about 2000 daltons. The number of PEG side chains of more than 90% of monomer units of the specified main chain of the polymer. Also included are many of the side groups of cisplatin or oxaliplatin, dissociatio associated with the main circuit. The number of side groups of cisplatin or oxaliplatin ranges from 25% to 75% of the number of monomer units of the main polymer chain.

Still another aspect of the invention is directed to biocompatible conjugated polymer nanoparticles including a main chain of poly(isobutylene-alt-maleic acid). The primary circuit comprises from 25 to 50 monomers. Also included are many of the side chains of glucosamine, covalently associated with the specified main chain. The number of side chains glucosamine is from 50% to 100% inclusive the Monomeric units of the specified main chain of the polymer. Also included are many of the side groups of cisplatin or oxaliplatin, dissociatio associated with the main circuit. The number of side groups of cisplatin or oxaliplatin ranges from 25% to 75% the number of monomer units of the main polymer chain.

Another aspect of the invention is directed to biocompatible conjugated polymer nanoparticles including a main chain of poly(isobutylene-alt-maleic acid). The main chain contains from 25 to 50 monomers. Also included are many of the side chains of glucosamine, covalently associated with the specified main chain. The number of side chains glucosamine more than 75% of monomer units of the specified main chain of the polymer. Also included are many of the side groups of cisplatin or oxaliplatin, dissociatio associated with the main circuit. The number of side groups of cisplatin or oxaliplatin ranges from 25% to 75% of the number of monomer units of the main polymer chain.

Still another aspect of the invention is directed to nanoparticles conjugated with complex carboxylic acid-platinum II (Pt(II)), including the carboxylic acid-(Pt(II)) and many lipid-polymer chains. Carboxylic some of the listed complex of carboxylic acid-cisplatin/oxaliplatin is covalently associated with the specified whether the ID of the polymer chains.

Another aspect of the invention is directed to vesicular, micellar or liposomal connection involving many nanoparticles formula, as described here.

Still another aspect of the invention is directed to pharmaceutical compositions, comprising any of nanoparticles or compounds described herein, and a pharmaceutically acceptable carrier.

Still another aspect of the invention is directed to a method of treating cancer or metastasis. The method involves selecting a subject in need of treatment of cancer or metastasis, and introduction to the subject an effective amount of any of the nanoparticles, the compounds or compositions described herein.

List of figures

Figure 1 represents a scheme of the synthesis of PMA-Cisplatin. Load different amounts of cisplatin on the polymer affects the size of the nanoparticles, as measured by the DLS method, or THEMES. Shows a graph of the cytotoxicity studies of media polyisobutylene-maleic acid (PIMA) 2 and conjugate (RMA-CISPLATIN) (6).

Figure 2 shows a scheme of the derivatization PMA using EDA. Derivationally polymer used for the synthesis of complex cisplatin. The graph shows the effect of different treatments on cell viability LLC after 48 hours incubation.

Figure 3 shows a scheme of the synthesis of PMA-GA-Cisplatin. The formation of the complex with cisplatin was performed within 48 hours. This caused the formation nanocasting size in the range of about 100 nm, as you can see from DLS measurements.

Figure 4. The left graph shows the number of active cisplatin, which is released from the nanoparticles PMA-GA-Cisplatin at incubation with lysate LLC. The graph on the right the concentration-effect" shows the effect of different treatments on cell viability Lewis lung carcinoma during incubation with the active agents within 48 hours. Cell viability was measured using MTS assay.

Figures 5A-5E are linear graphs (Fig.5A and 5B) and the histograms (Fig.5C-5E), showing the effectiveness and toxicity profile of free cisplatin and cisplatin nanoparticles in a model of Lewis lung carcinoma. Tumors induced by injection of LLC cells to mice c57/BL6. Demonstrates the effect of treatment on tumor volume (Fig.15A) and body weight (Fig.5B) during the treatment period. The animal was injected three times the dose (shown by arrows on the X axis). Data are presented as mean±STD.off., n=4-8. Also shows the effect of treatment on the weight of the kidneys (Fig.5C) and spleen (Fig.5D) as a marker of nephrotoxicity and hematologic toxicity (n=4-6). The image at the top of each graph show illustrative organs from each of the treated groups. Fig.5E shows the biodistribution Pt in the kidneys and tumors measured using ICP spectroscopy within 24 hours after the conduct of free cisplatin and the and nanoparticles of cisplatin (cisplatin dose equal to 8 mg/kg).

Figure 6 is a scheme showing the synthesis of PMA-GA-Cisplatin (8).

Figure 7. The effect of PMA-GA-Cisplatin in the Lewis lung carcinoma. Cells were incubated for 48 hours with drugs or media and then tested for viability using the MTS assay.

Figure 8. Effect of different treatments on tumor growth and body weight in vivo. Tumors induced by injection of LLC cells to mice c57/BL6.

Figure 9 is a graph showing the amount of platinum loaded in the polymer, which was calculated using the method of UV-VIS spectroscopy.

Figure 10. Scheme showing the synthesis of the lipid complex maleic acid-cisplatin, which can form micelles in water.

Figures 11A and 11B are diagrams showing the construction of the nanoparticles of cisplatin-based SAR. Fig.11A shows the mechanism underlying the intracellular activation of cisplatin by hydrating. Leaving groups of cisplatin and its analogues (shown by blue lines) replaced IT prior to DNA binding. Fig.11B shows the chemical synthesis PIMA-cisplatin and complex PIMA-glucosamine (PIMA-GA-cisplatin. Transformation polymolecular anhydride (n=40) (I) primulinum acid [PIMA] (2) makes it possible complexation [NH2]2Pt[OH]2through dicarboxylato communication (6). The derivatization one is about fragment PIMA glucosamine (4) and complex formation [NH 2]2Pt[OH]2can lead to the formation of two isomers (8) and (10), depending on pH, which is characterized by a unique NMR spectra of Pt (Fig.11B).

Figures 12A and 12B are characteristic of nanoparticles of cisplatin. Increasing the amount of Pt on the main circuit PIMA (n=40) increases the size of the formed nanoparticles. At the optimum ratio of Pt to the polymer, the inventors obtained nanoparticles of less than 150 nm, the size of the clipped below, which makes it possible preferential homing in the tumor. In Fig.12A shows that when derivatization of all of monomer units PIMA glucosamine and sequential complexation with Pt nanoparticles are formed less than 150 nm. In Fig.12B shows the total amount of platinum loaded on mg of the polymer at the given value.

Fig.13A-13TH are line graphs showing the in vitro characterization of nanoparticles of cisplatin. In Fig.13A and 13B shows the dependence of the concentration-effect of different treatments on cell viability as measured by MTS assay. The X-axis shows the equivalent concentration of cisplatin. When using polymer controls the dose of polymer used was equivalent to the dose that is used to deliver this specific doses of cisplatin in a comprehensive form. PIMA was also derivatization Ethylenediamine formed for the I PIMA-EDA, that provides an environment for the formation of a complex with platinum, similar PIMA-GA. Unlike PIMA-GA, PIMA-EDA shows internal toxicity. PIMA-GA-Cisplatin [sour] refers to the isomer formed in the acidic environment of complex formation, while PIMA-GA-Cisplatin [alkaline] refers to the isomer formed in an alkaline environment. In Fig.13C-13TH illustrates the effects of nanoparticles PIMA-GA-Cisplatin on the viability of LLC cells, when the main circuit PIMA (40 of monomer units) is derivateservlet to varying degrees. PIMA-GA-Cisplatin contains from 30 to 40 Monomeric units, derivatizing glucosamine, while all monomer units PIMA-GA-40 and PIMA-GA-200 are derivationally. [a] and [b] are isomers formed in acidic and alkaline environments, when the polymers form a complex with cisplatin. Table 1 presents the corresponding IC50 values.

Figures 14A-14J are FACS image (Fig.14A-14N) and histograms (Fig.14I and 14J), showing that the processing of the PIMA-GA-Cisplatin induces cell death. Illustrative FACS image cells T (Fig.14A-14D) and LLC (Fig.14-14N) show the percentage in each quadrant after treatments free cisplatin or cisplatin nanoparticles. Carboplatin was used as control for comparison (Fig.14D and 14N). Cells were incubated with drugs for 24 h, the donkey which were labeled Annexin-V FITC and stained propidium the iodide.

Figure 15 is a diagram showing the tagging polymer PIMA-GA using FITC to monitor the cellular uptake of the nanoparticles.

Figure 16 is a line graph showing the effect of pH and environment complexation of Pt on the kinetics of release. The nanoparticles were incubated at pH 5.5 or pH 8.5 in a dialysis bag and quantify the release over time. Used nanoparticles [PIMAGA-Cisplatin (O->Pt)] were generated by formation of a complex of the polymer and cisplatin in conditions of acidic pH [6.4], except PIMA-GA-Cisplatin (Pt->N), when the formation of the complex was carried out under conditions of alkaline pH with the formation of stable isomer [PIMA-GA-Cisplatin (N->Pt)]. Data are presented as mean±STD. off. from n=3.

Figures 17A-17D are line graphs (Fig.17A and 17 b) and histogram (Fig.17C and 17D), which shows that the nanoparticles PIMA-GA-cisplatin are similar antitumor activity with reduced systemic toxicity compared to free cisplatin in models of breast cancer C. Line graphs show the effect of treatment on tumor volume (Fig.17A) and body weight (Fig.17B) during the treatment period. The animal was injected three times the dose (shown by arrows on the X axis). Data are presented as mean±STD. off., n=4-8. Histograms show de is the effect of treatment on the weight of the spleen (Fig.18C), and kidney (Fig.17D) as a marker of nephrotoxicity and hematologic toxicity (n=4-6)*P< 0.05 compared with the group treated with carrier [ANOVA followed a posteriori comparisons criterion Newman of Tulsa]. Carboplatin [3 mg/kg] was used as a control.

Figures 18A and 18B represent the histogram (Fig.18A) and line graph (Fig.18V), which shows the inhibition of nanoparticles PIMA-GA-cisplatin tumor growth in a model of K-rasLSL/PtenfI/fIof ovarian cancer. As can be seen in Fig.18A, quantification of bioluminescence showed significantly reduced the signal from luciferase tumors of mice treated with cisplatin nanoparticle compared to vehicle (p<0.05, one-factor analysis of variance ANOVA). In Fig.18B shows the toxicity of the drugs evaluated by measuring the total mass of the body. Daily recording of body weight showed a significant loss of body weight in the group treated with free cisplatin compared with both groups treated with cisplatin nanoparticle (1.25 mg/kg and 3 mg/kg) (P<0.05, two-factor analysis of variance ANOVA).

Figures 19A and 19 b are histograms showing the distribution of Pt after the introduction of cisplatin, cisplatin nanoparticles [PIMA-GA-Cisplatin (O->Pt)] or carboplatin in breast cancer or ovarian cancer. Processing was performed as described in Fig.17 and 18. The level of Pt in different tissues, taken after the autopsy, the quantities of the NGOs was estimated using mass spectrometry with inductively coupled plasma (ICP).

Figures 20A and 20B are diagrams showing SAR-inspired design of nanoparticles oxaliplatin. Figure 20A shows the mechanism underlying the intracellular activation oxaliplatin by hydrating. Figure 20B shows the chemical synthesis of complex PIMA - oxaliplatin and PIMA-glucosamine (PIMA-GA)-oxaliplatin. Oxaliplatin-IT can form a complex with PIMA through dicarboxylato links. The derivatization of a single piece of PIMA glucosamine and education complex with oxaliplatin can lead to the formation of two isomers, depending on the pH.

Figures 21A and 21B are line graphs showing the dependence of the concentration-effect of different treatments on cell viability as measured by MTS assay. For this study used a cell line, breast cancer cell lines cancer Lewis lung (Fig.21A) and T (Fig.21B). The X-axis shows the equivalent concentration of platinum. When using polymer controls the polymer dose was equivalent to the dose that is used to deliver a given dose oxaliplatin in a comprehensive form. PIMA-GA-ox refers to the isomer [PIMA-GA-Oxaliplatin (O->Pt)], formed in the acidic environment of complex formation. Curve PIMA-GA-oxaliplatin shifted to the left, which indicates that the higher the th performance of nanoparticles in relation to antitumor activity compared with free oxaliplatin.

Figures 22A-22E are line graphs (Fig.26A and 26C) and histograms (Fig.22S-22E), which shows that the nanoparticles PIMA-GA-oxaliplatin have shown similar anti-tumor effect with reduced systemic toxicity compared to free oxaliplatin in models of breast cancer C. Line graphs show the effect of treatment on tumor volume (Fig.22A) and body weight (Fig.22) during the treatment period. Animals were injected dose three times. Data are presented as mean±STD. off., n=4-8. Histograms show the effect of treatment on tumor weight (Fig.22 ° C), kidney (Fig.22D) and spleen (Fig.22E) as a marker of nephrotoxicity and hematologic toxicity (n=4-6).

Figure 23 is a line graph showing zavisimosti "concentration-effect of different complexes oxaliplatin on cell viability as measured by MTS assay.

Figure 24 is a line graph showing the effect of cisplatin, carboplatin and PIMA-GA-200(A) cell viability.

Figure 25A is a diagram showing the synthesis of the conjugate is cholesterol-succinic acid and the formation of the complex Pt conjugate.

Figure 25 shows the dynamic scattering of laser radiation of lipogenesis. The size of lipogenesis is less than 150 nm.

Figure 26 is a linear the graph, showing the kinetics of release of Pt from lipogenetic over time and the influence of pH. The rate of release is higher in acidic pH conditions corresponding to the acidic pH inside the tumor, which contributes to selective release of active platinate in the tumor.

Figures 27A-27C are line graphs showing the effect of lipogenetic cisplatin on cell viability T breast cancer. Cell survival was measured using the MTS assay. Processing lebanonization resulted in rapid cell death within 12 hours compared with cisplatin or carboplatin (Fig.27A). It was found that in all three time lipogenetic cisplatin was more effective than cisplatin. Carboplatin is the least effective of all tested platinate (Fig.27A-27C).

Figures 28A and 28C are line graphs showing the effect of lipogenetic cisplatin on the viability resistant to this drug called cisplatin cell line (SR) hepatocellular cancer and cell line Lewis lung cancer (LLC). Cisplatin acts on SR only at the highest concentration, whereas cells are susceptible to lipogenetic cisplatin (Fig.28A). Carboplatin has no effect in this range of concentrations (Fig.28A and 28C). Lipogenetic cisplatinbased anticancer effect of higher than free cisplatin in LLC (Fig.28VDC).

Figures 29A-E are line graphs (Fig.29A and 29B) and histograms (Fig.29S-E), which shows the effectiveness of lipogenetic cisplatin in a synergistic model of tumor E in vivo. Line graphs show the effect of different treatments on tumor growth (Fig.29A and 29S) and body mass (as a marker of systemic toxicity, Fig.29V). Histograms show the mass of the kidneys (Fig.29D), and spleen (Fig.E) as markers of nephrotoxicity and hematological toxicity. As can be seen from the figures, liyanarachchi cisplatin induced greater antitumor activity with reduced systemic and nephrotoxicity.

The implementation of the invention

The invention is directed to biocompatible conjugated polymer nanoparticles including a main chain of the copolymer, many of the side chains covalently associated with the specified main chain, and a lot of platinum compounds, dissociatio associated with the specified main chain. In General, compounds of platinum connected to the main chain through communication with the side chains.

In one embodiment, the copolymer comprises monomers of maleic acid.

In a preferred embodiment, the copolymer is a poly(isobutylene-alt-maleic acid) (PIMA or PMA).

In some embodiments, the copolymer contains from 2 to 100 monomer units. In some embodiments, the copolymer contains from 25 to 50 Monomeric units.

In some embodiments, the side chains are selected from the group consisting of polymers, sugars, carboxylic acids, dicarboxylic acids, amides and combinations thereof.

In preferred embodiments, the side chain is a polyethylene glycol (PEG). Side chains of PEG can be represented by the formula-C(O)-NH-PEG.

In some embodiments, PEG side chains have a molecular weight of from 100 to 5000 daltons. In some embodiments, PEG side chains have a molecular weight of from 1000 to 3000 daltons. In a preferred embodiment, PEG side chains have a molecular weight of about 2000 daltons.

In some embodiments, the side chains are monosaccharides. In a preferred embodiment, the monosaccharide is glucosamine. Monosaccharide side chains can be represented as-C(0)-saccharide.

In the invention can be any of the platinum compounds. Preferably, a platinum compound was a compound of platinum (II) or platinum (IV). In some embodiments, the platinum compound (II) is selected from the group consisting of cisplatin, oxaliplatin, carboplatin, paraplatin, satraplatin and combinations thereof. In a preferred embodiment, the side group of compounds of platinum (II) is cisplatin or oxaliplatin.

In some embodiments, the platinum compound (II) is selected from the

group, vkluchaya the Pt(NH 3)2Pt(NH3)(2-methylpyridine), andwhere p is 0-3. In a preferred embodiment, a platinum compound (II) is Pt(NH3)2.

In some embodiments, the platinum compound (II) iswhere p is 0-3.

In some embodiments, the platinum compound (II) includes at least two nitrogen atom, the nitrogen atoms are directly connected with platinum. In another embodiment, two nitrogen atom connected to each other via the optional substituted linker, such as acyclic or cyclic linker. Cyclic linker means binding fragment that contains at least one ring structure. Cyclic linkers can be aryl, heteroaryl, cilil or heterocyclyl.

In some embodiments, at least one nitrogen atom, which is connected with platinum, is a ring atom heteroaryl or heterocyclyl. In a preferred embodiment, heteroaryl is an optional substituted pyridine, such as 2-methylpyridin.

In some embodiments, many of the side chains corresponds to the number of from 50% to 100% inclusive of the number of monomer units of the specified polymer main chain. This means that from 50% to 100% of the Monomeric units have at least one side chain that is associated with Monomeric unit. On the quantity side chains may be more than the total number of monomer units. For example, two of the side chain can be attached to the monomer of maleic acid.

In some embodiments, many of the side chains corresponds to the number of more than 90% of the number of monomer units specified the main polymer chain.

In some embodiments, the set of platinum compounds corresponds to the number of from 10% to 100% inclusive of the number of monomer units specified the main polymer chain. In General, there is a one-to-one correspondence between the platinum compounds and Monomeric subunits. Thus, the percentage refers to the ratio of the number of monomer units, which are connected with a platinum compound, to the total number of monomer units present in the main polymer chain.

In some embodiments, the set of platinum compounds corresponds to the number from 25% to 75% of the number of monomer units specified the main polymer chain.

In General, from 10 to 500 μg of platinum compounds can be loaded with 1 mg of the polymer. Preferably, from 50 to 250 μg, more preferably from 150 to 200 µg compounds of platinum was loaded on 1 mg of polymer. In some embodiments, the inventors have received a load of 175±5 µg/mg polymer.

In some embodiments, the side chains contain dicarboxylic acid. In some var is the ants dicarboxylic acids have the formula HOOC-R-COOH, where R represents a C1-C6alkyl, C3-C6alkenyl or2-C6quinil. In a preferred embodiment, the dicarboxylic acid is maleic acid.

In some embodiments, the copolymer contains at least one monomer having the formula-CH(CO2H)-R-CH(C(O)R')-, where R represents a bond, C1-C6alkylen where alkylene may include one or more double or triple links; and R' is a substituted nitrogen atom. Preferably, R represents a connection.

In some embodiments, from about 50% to 100% inclusive Monomeric subunits in the main polymer chain represent-CH(CO2H)-R-CH(C(O)R')-, where R represents a bond, C1-C6alkylen where alkylene may include one or more double or triple links; and R' is optional substituted by a nitrogen atom.

In some embodiments, at least 90% or more Monomeric subunits in the main polymer chain represent-CH(CO2H)-R-CH(C(O)R')-, where R is a bond, C1-C6alkylene, with alkylene may include one or more double or triple links; and R' is a substituted nitrogen atom.

In some embodiments, the copolymer includes at least one monomer having the formula-CH(CO2H)-R-CH(C(O)R')CH2C(Me2)- or-CH(C(O)R')-R-CH(CO2H)- CH2 With(Me2)-, where R is a bond, C1-C6alkylene, with alkylene may include one or more double or triple links; and R' is a substituted nitrogen atom. Preferably, R represents a connection.

In some embodiments, the copolymer contains from 50% to 100% inclusive of monomers having the formula-CH(CO2H)-R-CH(C(O)R')CH2C(Me2)- or-CH(C(O)R')-R-CH(CO2H)-CH2C(Me2)-, where R is a bond, C1-C6alkylene, with alkylene may include one or more double or triple links; and R' is a substituted nitrogen atom.

In some embodiments, the copolymer contains at least 90% of monomers having the formula-CH(CO2H)-R-CH(C(O)R')CH2C(Me2)- or-CH(C(O)R')-R-CH(CO2H)-CH2With(Me2)-, where R is a bond, C1-C6alkylene, with alkylene may contain one or more double or triple links; and R' is a substituted nitrogen atom.

In one embodiment, R' representsor NH(CH2CH2O)mCH where m is 1-150.

In some embodiments, at least one monomer of the polymer contains two side chains selected from the group consisting of carboxylic acid, amide and ether complex. These side chains are separated from each other 1, 2, 3, 4, 5, 6, 7, 8, 9 or ten atoms of carbon, oxygen, nitrogen, sulfur is whether their combination. Preferably, the above amide and ether side chains were separated from each other by two carbon atoms. Preferably, at least one of the side chains was not a carboxylic acid.

In some embodiments, at least one monomer of the polymer contains two side chain carboxylic acid. These side chain carboxylic acid is separated from each other 1, 2, 3, 4, 5, 6, 7, 8, 9 or ten atoms of carbon, oxygen, nitrogen, sulfur, or a combination. Preferably, these side chain carboxylic acids were separated from each other by two carbon atoms. These carbon atoms may be linked to each other by means of a simple or double bonds.

In some embodiments, at least one monomer of the polymer contains a side chain carboxylic acid and amide. These side chain carboxylic acid and amide separated from each other 1, 2, 3, 4, 5, 6, 7, 8, 9 or ten atoms of carbon, oxygen, nitrogen, sulfur, or a combination. Preferably, these side chain carboxylic acid and amide were separated from each other by two carbon atoms. These carbon atoms may be linked to each other by means of a simple or double bonds.

In some embodiments, at least one monomer of the polymer contains a side chain carboxylic acid and hard what about the ether. These side chain carboxylic acid of ester are separated from each other 1, 2, 3, 4, 5, 6, 7, 8, 9 or ten atoms of carbon, oxygen, nitrogen, sulfur, or a combination. Preferably, these side chain carboxylic acid and ether complex were separated from each other by two carbon atoms. These carbon atoms may be linked to each other by means of a simple or double bonds.

In some embodiments, at least one monomer of the polymer contains a side chain amide and ether complex. These side chain amide and ether complex are separated from each other 1, 2, 3, 4, 5, 6, 7, 8, 9 or ten atoms of carbon, oxygen, nitrogen, sulfur, or a combination. Preferably, these side chain amide and ether complex were separated from each other by two carbon atoms. These carbon atoms may be linked to each other by means of a simple or double bonds.

In some embodiments, at least one monomer of the polymer contains two side-chain amide. These side chain amide separated from each other 1, 2, 3, 4, 5, 6, 7, 8, 9 or ten atoms of carbon, oxygen, nitrogen, sulfur, or a combination. Preferably, these side chain amide and ether complex were separated from each other by two carbon atoms.

In some embodiments, at least one monomer poly the EPA has two side chains of ester. These side chains of ester are separated from each other 1, 2, 3, 4, 5, 6, 7, 8, 9 or ten atoms of carbon, oxygen, nitrogen, sulfur, or a combination. Preferably, these side chain amide and ester were separated from each other by two carbon atoms.

In some embodiments, the polymer includes two side chains selected from the group consisting of carboxylic acid, amide and ether complex. These side chains are separated from each other 1, 2, 3, 4, 5, 6, 7, 8, 9 or ten atoms of carbon, oxygen, nitrogen, sulfur, or a combination. Preferably, these side chain amide and ether complex were separated from each other by two carbon atoms. Preferably, at least one of the side chains was not a carboxylic acid.

In some embodiments, the polymer contains at least two side chain carboxylic acid. These side chain carboxylic acid is separated from each other 1, 2, 3, 4, 5, 6, 7, 8, 9 or ten atoms of carbon, oxygen, nitrogen, sulfur, or a combination. Preferably, these side chain carboxylic acids were separated from each other by two carbon atoms.

In some embodiments, the polymer contains a side chain carboxylic acid and amide. These side chain carboxylic acid and amide side chains are separated from each other 1, 2, 3, 4, 5, 6, 7, 8, 9 or on satu carbon atoms, oxygen, nitrogen, sulfur, or a combination. Preferably, these side chain carboxylic acid and amide were separated from each other by two carbon atoms.

In some embodiments, the polymer contains a side chain carboxylic acid and a complex ester. These side chain carboxylic acid of ester are separated from each other 1, 2, 3, 4, 5, 6, 7, 8, 9 or ten atoms of carbon, oxygen, nitrogen, sulfur, or a combination. Preferably, these side chain carboxylic acid and ether complex were separated from each other by two carbon atoms.

In some embodiments, the polymer contains a side chain amide and ether complex. These side chain amide and ether complex are separated from each other 1, 2, 3, 4, 5, 6, 7, 8, 9 or ten atoms of carbon, oxygen, nitrogen, sulfur, or a combination. Preferably, these side chain amide and ether complex were separated from each other by two carbon atoms. These carbon atoms may be linked to each other by means of a simple or double bonds.

In some embodiments, the polymer contains two side-chain amide. These side chain amide separated from each other 1, 2, 3, 4, 5, 6, 7, 8, 9 or ten atoms of carbon, oxygen, nitrogen, sulfur, or a combination. Preferably, these side chain amide and ether complex were a Department of the us from one another by two carbon atoms.

In some embodiments, the polymer contains two side chains of ester. These side chains of ester are separated from each other 1, 2, 3, 4, 5, 6, 7, 8, 9 or ten atoms of carbon, oxygen, nitrogen, sulfur, or a combination. Preferably, these side chain amide and ether complex were separated from each other by two carbon atoms.

The size of the nanoparticles of the invention may vary in the range of 25-250 nm, preferably 50-200 nm, more preferably 80-160 nm and most preferably 90-110 nm. Without being bound to any theory, the predominant homing nanoparticles with a size in the range of 80-160 nm in the tumor occurs as a result of increased permeability and detention. See, for example, Moghimi, et al., Pharmacol Rev. 2001 Jun; 53(2):283-318.

In some embodiments, the platinum compound dissociatio associated with the specified main chain through at least one coordination. Without being bound to any theory, coordination is more accessible and thus more easily releases the platinum compound.

In some embodiments, the binding of platinum compounds with the main chain of the biopolymer, in addition, includes carboxylato communication. In some embodiments, the platinum compound is related to the main chain through coordination and carboxylato connection.

Must be ponat is, despite the description of the relationship with the main chain, experienced in this field specialist will be apparent that the platinum compound, in General, associated with one or more side chains, which is itself linked to the main chain. Thus, any description of the binding of platinum compounds with the main circuit includes the cases when the platinum compound is associated with the side chain, which, in addition, linked to the main chain.

In some embodiments, the coordination relationship is between the platinum atom of the platinum compounds and the oxygen of the side chain. Preferably, coordinating the relationship was between the platinum and the carbonyl oxygen.

In some embodiments, the coordination relationship is between the platinum atom of the platinum compounds and amide oxygen of the side chain. In some embodiments, the coordination relationship is between the platinum atom of the platinum compounds and carbonyl oxygen of ester side chain.

In some embodiments, the copolymer contains at least one monomer of maleic acid, in which at least one carboxylic acid mentioned at least maleic acid is derivational in amide.

In some embodiments, from about 50% to 100% inclusive of monomer units in the main polymer chain are monomer of maleic acid and at least one carboxylic acid of the specified monomer of maleic acid is derivational in amide.

In some embodiments, at least 90% of monomer units in the main polymer chain are monomer of maleic acid and at least one carboxylic acid of the specified monomer of maleic acid is derivational in amide.

The loading of platinum compounds on the polymer can be represented as the percentage of mg of platinum compounds on mg polymer. For example, on the polymer PIMA-GA can be loaded to a maximum of 0.375 mg of cisplatin, so that the load in 37.5% represents the maximum load for this particular polymer. The load may vary from about 1% to theoretical total load for the polymer.

In some embodiments, the loading of platinum compounds is 1%-37.5%. Interest burden is mg of platinum compounds associated with mg polymer.

In some embodiments, the loading of platinum compounds is 1%-6%. In some embodiments, the communication overhead of Pt(II) is from 0.01% to 1%.

Another aspect of the invention is directed to biocompatible conjugated polymer nanoparticles including a main chain of poly(isobutylene-alt-maleic acid). The primary circuit contains 25-50 monomers. Also included are many of the side chains of PEG covalently connected to the specified main chain. Side chains of PEG have a molecular weight of from 1000 to 3000 daltons. The number of PEG side chains is from 50% to 100% inclusive of the number of monomer units of the main polymer chain. Also included are many of the side groups of cisplatin, dissociatio associated with the main circuit. The number of side groups of cisplatin is from 25% to 75% of the number of monomer units of the main polymer chain.

Still another aspect of the invention is directed to biocompatible conjugated polymer nanoparticles including a main chain of poly(isobutylene-alt-maleic acid). The primary circuit consists of 40 monomers. Also included are many of the side chains of PEG covalently associated with the specified main chain. Side chains of PEG have a molecular weight of 2000 daltons. The number of PEG side chains of more than 90% of monomer units of the specified main chain of the polymer. Also included are many of the side groups of cisplatin, dissociatio associated with the main circuit. The number of side groups of cisplatin is from 25% to 75% of the number of monomer units of the main polymer chain.

Still another aspect of the invention is directed to biocompatible conjugated polymer nanoparticles including a main chain of poly(isobutylene-alt-maleic acid). The primary circuit comprises from 25 to 50 monomial is the moat. Also included are many of the side chains of glucosamine, covalently associated with the specified main chain. The number of side chains glucosamine is from 50% to 100% inclusive of monomer units specified the main polymer chain. Also included are many of the side groups of cisplatin, dissociatio associated with the main circuit. The number of side groups of cisplatin is from 25% to 75% of the number of monomer units of the main polymer chain.

Another aspect of the invention is directed to biocompatible conjugated polymer nanoparticles including a main chain of poly(isobutylene-alt-maleic acid). The primary circuit comprises from 25 to 50 monomers. Also included are many of the side chains of glucosamine, covalently associated with the specified main chain. The number of side chains glucosamine more than 90% of monomer units of the specified main chain of the polymer. Also included are many of the side groups of cisplatin, dissociatio associated with the main circuit. The number of side groups of cisplatin is from 25% to 75% of the number of monomer units of the main polymer chain.

Still another aspect of the invention is directed to conjugated nanoparticles of complex carboxylic acid is a compound of platinum, including the carboxylic acid is Obedinenie platinum and many lipid-polymer chains. Carboxylic some of the listed complex of carboxylic acid-platinum compound is covalently associated with these lipid-polymer chains.

In a preferred embodiment, the carboxylic acid is maleic acid. In some embodiments, the polymer is PEG.

In certain embodiments, the loading of platinum compounds is 1%-37.5%. In certain embodiments, the loading of platinum compounds is 1%-6%.

The platinum compound can be a compound Pt(II) or the compound Pt(IV). In some embodiments, the compound Pt(II) is selected from the group consisting of cisplatin, oxaliplatin, carboplatin, paraplatin, satraplatin and combinations thereof. In a preferred embodiment, the compound Pt(II) is cisplatin.

Another aspect of the invention is directed to a compound in the form of vesicles, micelles or liposomes, including many of the nanoparticles according to the formula, as described here.

Still another aspect of the invention is directed to a pharmaceutical composition comprising any of nanoparticles or compounds described herein, and a pharmaceutically acceptable carrier.

Still another aspect of the invention is directed to a method of treating cancer or metastasis. The method involves selecting a subject in need of treatment of cancer or metastasis, and the introduction of an effective amount of any of the nanoparticles, compounds or compositions, opican the x here.

In some embodiments, the cancer or metastasis is selected from the group consisting of susceptible or resistant to platinum tumors, including tumors of the breast, head and neck, ovary, testis, pancreas, oral esophagus, gastrointestinal tract, liver, gall bladder, lung, melanoma, skin cancer, sarcoma, malignant diseases of the hematopoietic system, brain tumors, including glioblastoma, and tumors of neuroectodermal origin.

In still another aspect, the invention provides a method for preparation of polymeric nanoparticles of platinum compounds, the method includes conjugating compounds of platinum with a biocompatible polymer or biocompatible copolymer. Without being bound to theory, conjugation of platinum compounds with biocompatible polymer under conditions of acidic pH leads to the formation of nanoparticles, which are more active in vivo than in the case when the conjugation is carried out at alkaline pH.

Thus, in some embodiments, the conjugation is performed at a pH of below 7, preferably, the pH was in the range of from 1 to 6.9. In some preferred embodiments, the conjugation was carried out at pH 6.5.

The inventors have observed that conjugation in alkaline conditions promote formation of isomeric to the Plex PIMA-GA-Cisplatin with monocarboxylate and more stable Pt< ->N coordination bond. On the contrary, the formation of the complex PIMA-GA and cisplatin in conditions of acidic pH creates an isomeric state, characterized by monocarboxylate communication and Pt<->O coordination bond. Thus, the conditions of conjugation, which lead to the formation of Pt<->O coordination through Pt<->N coordination bond, are preferred for conjugation.

In General, for a polymer used an excess of compounds of Pt(II). In some embodiments, for the polymer used 5-25 moles of excess compounds of Pt(II). Preferably used for the polymer 10-20 moles of excess compounds of Pt(II). In one preferred embodiment, for polymer used 15 moles of excess compounds of Pt(II).

In still another aspect, the invention provides dicarbonyl molecule associated with the lipid molecule. This connection can be represented by the structure of the lipid-linker-dicarbonyl. These molecules can be used for the formation of a complex with platinum compounds such as cisplatin, oxaliplatin or other platinate and platinum compounds described herein, by carboxylato communication and/or coordination bonds. These compounds may be mixed with the appropriate lipids/phospholipids in nanoparticles smaller than che is 150 nm, which release Pt pH-dependent manner. Once completed, these nanoparticles show improved efficacy and toxicity profile compared with carboplatin and cisplatin and are active in resistant to this drug called cisplatin cancer.

These nanoparticles can be prepared with the inclusion of pharmaceutically active agents for delivery.

The term "Lipid" is used in the ordinary way to refer to molecules that are soluble to a greater or lesser extent in organic solvents, such as alcohols, and relatively insoluble in water. Thus, the term "lipid" includes compounds with varying chain length from short, containing 2 carbon atoms, to long, containing about 28 carbon atoms. In addition, the compounds can be saturated or unsaturated, and in the form of straight or branched chains, or in the form of unfused or condensed ring structures. Examples of lipids include, but are not limited to, fats, waxes, sterols, steroids, bile acids, fat-soluble vitamins such as A, D, E and K), monoglycerides, diglycerides, phospholipids, glycolipids, sulfolipids, linoleamide, chromalife (lipochrome), glycerophospholipids, sphingolipids, prenol-lipids, charalabidis, are polyketides and fatty acids. In some embodiments, the lipid is cholesterol or distearoylphosphatidylcholine.

In General, can be any molecule that contains two carbonyl groups. In some embodiments dicarbonyl molecule is a dicarboxylic acid or keto-carboxylic acid. In some preferred embodiments dicarbonyl molecule is succinic acid.

In some embodiments dicarbonyl molecule, R is OC(O)-R,-C(O)-, where R represents a C1-C6alkylen, with alkylene may contain one or more double or triple links, and/or the main chain alkylene may be interrupted by one or more of O, S, S(O), SO2, NH, C(O); and R' represents H, alkyl, alkenyl, quinil, aryl, heteroaryl, heterocyclyl, each of which may be optional substituted. Preferably, R represents a CH2, -CH2CH2-, -CH2CH2-CH2- or CH=CH-. Preferably, R' represents N.

Dicarbonyl molecule can be linked to a lipid molecule directly or through a linker molecule. The term "linker" means an organic fragment that connects two parts of the connection. Linkers typically comprise a direct bond or an atom such as oxygen or sulfur, link, such as NH, C(O), C(O)NH, SO, SO2, SO2NH, or a chain of atoms, such as substituted or unsubstituted alkyl, substituted or unsubstituted of alkenyl, C is displaced or unsubstituted quinil, arylalkyl, arylalkyl, arylalkyl, heteroallyl, heteroaromatic, heteroallyl, geterotsiklicheskikh, heterocyclisation, geterotsiklicheskikh, aryl, heteroaryl, heterocyclyl, cycloalkyl, cycloalkenyl, alkylacrylate, alkylresorcinol, alkylresorcinol, alkenylsilanes, alkenylsilanes, alkenylsilanes, alkynylaryl, alkenylsilanes, alkenylsilanes, alkylchlorosilanes, alkylchlorosilanes, alkylchlorosilanes, alkenylsilanes, alkenylsilanes, alkenylsilanes, alkynylaryl, alkynylpyrazolyl, alkenylsilanes, alkylchlorosilanes, alkylchlorosilanes, alkylchlorosilanes, alkynylpyrazolyl, alkynylpyrazolyl, alkynylpyrazolyl, alginolyticus, alkynylpyrazolyl, alkynylpyrazolyl, alkylaryl, alkynylaryl, alkynylaryl, alkylether, alkenylamine, alkynylaryl, where one or more metileno can be interrupted or terminated O, S, S(O), SO2, NH, C(O). It should be understood that dicarbonyl molecule and/or lipid can be modified to include functional groups for binding with each other or with the linker.

In some embodiments, the linker is a diamine, such as Ethylenediamine. In some Islands Ianto linker is PEG-NH 2.

In one preferred embodiment, the linker is-NHCH2CH2C(O)-. In another preferred embodiment, the linker is-CH2CH2NHC(O)-[OCH2CH2]z-NH-where z is 1-50. It is preferable that z was equal to 45.

In some embodiments, the lipid-dicarbonyl compound is such that shown in Fig.10 (compound 2) and 25 (connection 5).

In another aspect the invention provides a biocompatible polymer comprising at least one monomer having the formula-CH(CO2H)-R-CH(C(O)R')-, where R is a bond, C1-C6alkylene, with alkylene may contain one or more double or triple links; and R' is a substituted nitrogen atom. Preferably, R was a connection.

In some embodiments, the polymer contains from 2 to 100 monomer units having the formula-CH(CO2H)-R-CH(C(O)R')-, where R is a bond, C1-C6alkylene, with alkylene may contain one or more double or triple links; and R' is a substituted nitrogen atom.

In some embodiments, the polymer contains from 25 to 50 Monomeric units having the formula-CH(CO2H)-R-CH(C(O)R')-, where R is a bond, C1-C6alkylene, with alkylene may contain one or more double or triple links; and R' is a substituted nitrogen atom.

In some embodiments at% to 100% inclusive Monomeric subunits in the polymer main chain represent-CH(CO 2H)-R-CH(C(O)R')-, where R is a bond, C1-C6alkylene, with alkylene may contain one or more double or triple links; and R' is a substituted nitrogen atom.

In some embodiments, at least 90% or more Monomeric subunits in the polymer main chain represent-CH(CO2H)-R-CH(C(O)R')-, where R is a bond, C1-C6alkylene, with alkylene may contain one or more double or triple links; and R' is a substituted nitrogen atom.

In some embodiments, the copolymer includes at least one monomer having the formula-CH(CO2H)-R-CH(C(O)R')CH2C(Me2)- or-CH(C(O)R')-R-CH(CO2H)-CH2With(Me2)-, where R is a bond, C1-C6alkylene, with alkylene may contain one or more double or triple links; and R' is a substituted nitrogen atom. Preferably, R was a connection.

In some embodiments, the copolymer contains from 50% to 100% inclusive of monomers having the formula-CH(C02H)-R-CH(C(O)R')CH2C(Me2)- or-CH(C(O)R')-R-CH(CO2H)-CH2C(Me2)-, where R is a bond, C1-C6alkylene, with alkylene may contain one or more double or triple links; and R' is a substituted nitrogen atom.

In some embodiments, the copolymer contains at least 90% of monomers having the shape of the in-CH(CO 2H)-R-CH(C(O)R')CH2C(Me2)- or-CH(C(O)R')-R-CH(CO2H)-CH2With(Me2)-, where R is a bond, C1-C6alkylene, with alkylene may contain one or more double or triple links; and R' is a substituted nitrogen atom.

In some embodiments, R' is aOh or-NH(CH2CH2O)mCH3where m is 1-150.

These polymers can be used to generate nanoparticles and gels that can be used to deliver drugs. Thus, the invention also provides nanoparticles comprising a polymer described herein, and one or more bioactive agents ("bioactive agent").

Described here, the composition can be used in methods for sustained release of bioactive agents. In one embodiment, the method includes: (a) providing or introduction to the subject compositions described herein, the composition comprises a bioactive agent. Used here, the term "bioactive agent" refers to natural biological materials, for example materials of the extracellular matrix such as fibronectin, vitronectin and laminin; cytokines; growth factors and differentiation factors. "Bioactive agents" also belong to the artificially synthesized materials, molecules or compounds that about adut biological effects on biological cells, tissues or organs.

Suitable growth factors and cytokines include, but are not limited to, stem cell factor (SCF), granulocyte colony-stimulating factor (G-CSF), granulocyte macrophage-colony stimulating factor (GM-CSF), a growth factor, stromal cell-1, steel factor, vascular endothelial growth factor (VEGF), transforming growth factor - beta (TGFp), platelet growth factor (PDGF), angiopoietin (Ang), epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), hepaticotomy nuclear factor (HNF), nerve growth factor (NGF), bone morphogenetic protein (BMP), fibroblast growth factor (FGF), a growth factor for hepatocytes, insulin-like growth factor (IGF-1), interleukin (IL)-3, IL-1α, IL-1β, IL-6, IL-7, IL-8, IL-11 and IL-13, colony stimulating factors, thrombopoietin, erythropoietin, fit3 ligand and tumor necrosis factor-α (TNFα). Other examples are described in Dijke et A1., "Growth Factors for Wound Healing", Bio/Technology, 7:793-798 (1989); Mulder GD, Haberer PA, Jeter KF, eds. Clinicians' Pocket Guide to Chronic Wound Repair. 4th ed. Springhouse, PA: Springhouse Corporation; 1998:85; Ziegler, T. R., Pierce, G. F., and Herndon, D. N., 1997, International Symposium on Growth Factors and Wound Healing: Basic Science & Potential Clinical Applications (Boston, 1995, Serono Symposia USA), Publisher: Springer Verlag.

In some embodiments, suitable bioactive agents include, but without limitation, therapeutic agents. Used here, the term "therapeutic agent" refers to substances, which ispolzuetsa diagnosis, the treatment or prevention of disease. Any therapeutic agent known to a person skilled mid-level in this area as useful in the diagnosis, treatment or prevention of disease, is considered as a therapeutic agent in the context of the present invention. Therapeutic agents include pharmaceutically active compounds, hormones, growth factors, enzymes, DNA, plasmid DNA, RNA, siRNA, viruses, proteins, lipids, anti-inflammatory molecules, antibodies, antibiotics, anti-inflammatory agents, antisense nucleotides and transforming nucleic acids, or combinations thereof. Any of therapeutic agents may be combined, provided that such combination is biologically compatible.

Illustrative therapeutic agents include, but without limitation, those that can be found in Harrison''s Principles of Internal Medicine, 13thEdition, Eds. T. R. Harrison et al. McGraw-Hill, N. Y., NY; Physicians Desk Reference, 50thEdition, 1997, Oradell New Jersey, Medical Economics Co.; Pharmacological Basis of Therapeutics, 8thEdition, Goodman and Oilman, 1990; United States Pharmacopeia, The National Formulary, USP XII NF XVII, 1990; current edition of Goodman and Oilman''s The Pharmacological Basis of Therapeutics; and current edition of The Merck Index, the full contents of which are incorporated herein by reference.

Examples of therapeutic agents that can be included in the composition include, but without limitation, drug analytice the Kie of the medicinal product; gold salts; corticosteroids; hormones; antimalarials; indole derivatives; pharmaceutical agents for the treatment of arthritis; antibiotics, including tetracyclines, penicillin, streptomycin and aureomycin; anthelmintic and medicines against canine plague used for Pets and large animals, such as, for example, phenothiazines; medicines on the basis of sulfur, such as sulfisoxazole; anticancer drugs; pharmaceutical regulatory dependence tools, such as agents, controlling alcohol dependence and agents controlling nicotine addiction; antagonists of addiction to drugs such as methadone and drugs controlling weight; medicines that control the thyroid gland; analgesics; drugs, controlling fertilization or hormonal contraceptives; amphetamines; antihypertensive drugs; anti-inflammatory agents; antitussives; sedation; muscle relaxants; antiepileptic drugs; antidepressants; anti-arrhythmic drugs; vasodilator; protivogipertonicheskoe diuretics; antidiabetic agents; anticoagulants; antitubercular agents; ntipsihoticescoe agents; hormones and peptides. It should be clear that the above list is incomplete and only shows a wide variety of therapeutic agents that can be included in the composition. In some embodiments, therapeutic agent is mitoxantrone, protein (e.g., VEGF) or plasmid DNA.

The amount of therapeutic agent distributed in the composition depends on various factors, including, for example, a specific agent; function it will perform; the desired time period for release of the agent; the amount to be introduction. In General, the dosage of therapeutic agent, i.e., the amount of therapeutic agent in the composition is selected from the range of from about 0.001% (weight/weight)% to 95% (weight/weight), preferably from about 5% (weight/weight) to 75% (weight/weight) and more preferably from about 10% (weight/weight) to 60% (weight/weight).

Cisplatin [CIS-dichlorodiammineplatinum(P)] (CDDP) is an important class of anticancer agents and is widely used for the treatment of many malignant tumors, including tumors of the testis, ovary, cervix, head and neck and non-small cell lung cancer (Jamieson, et al, Chem. Rev. (1999), 99(9); 2467-2498). He also showed activity in triple negative breast cancer (Leong, et al., J. Clin. Invest. (2007), 117(5):1370-80). However, its application has datagramchannel, mainly because of nephrotoxicity or toxicity to the kidney (Madias, NE and Harrington, JT, Am. J.(1978), 65(2):307-14). To explore this limitation was highlighted in two directions, the first is aimed at the synthesis of analogues of platinum, the second on the designing of new systems nedostatki a way of delivering drugs directly to the site of the tumor. It is now established that the predominant homing nanoparticles with a size in the range of 80-120 nm in the tumor occurs as a result of the effect of the increased permeability and detention (EPR) (Moghimi, et al., Pharmacol. Rev. (2001), 53(2): 283-318). This can reduce systemic side effects and increased intratumoral delivery. It was found that nanoliposomal composition of cisplatin delivers in 50-200 times more of the drug in the tumor compared with the introduction of free cisplatin (Harrington, et al. Ann. Oncol. (2001) 12:493-496). Despite the presence of minimal toxicity, nanoliposomal composition has a moderate anti-tumor activity compared to cisplatin; reflecting problems not only delivery of platinum in a relatively inactive form, but the subsequent need to achieve significant release and activation within the tumor. The second strategy encapsulation of cisplatin in polymer systems had the problem because of its insolubility in organizes the x solvents and partial solubility in water, that led to a lack of load or inability to sustain a slow release. This necessitated the development of prodrugs of platinum (IV), which can be modified to increase the hydrophobicity and increase loading in the nanoparticles-based copolymer of polyactic-polyglycolide (Dhar et al., 2009). Or, cisplatin conjugatively with N-(2-hydroxypropyl)methacrylamide (NRMA) through peptidyl side chains and has been proved to its biological activity (Lin X, Zhang Q, Rice JR, Stewart DR, Nowotnik DP, Howell SB). Improved directional effects of chemotherapy based on platinum. Antitumor activity of platinum agent OR copolymer of NRMA in murine tumor models. Eur J Cancer. 2004 Jan; 40(2):291-7). However, such approaches require passing through enzymatic cleavage or intracellular recovery to activate the drug. Similarly, it was found that the complex FRAMES dendrimers platinum, which has increased the pressure medicines was 200-550 times less toxic than cisplatin, thanks to the strong ties formed between the polymer and Pt (Haxton KJ, Burt HM. Polymeric drug delivery of platinum-based anticancer agents. J Pharm Sci. 2009 Jul; 98(7):2299-316).

To construct nanoactive on the basis of cisplatin, which is not difficult, but solves the problems associated with these approaches, the inventors have combined newshouse information on biotransformation of cisplatin and understanding of the dependence of activity on the structure, which arose during the development of cisplatin analogues. Cisplatin is activated as a result of intracellular hydration of one or two leaving groups chloride with the formation of [Pt(NH3)2Cl(OH)2)]+and [Pt(NH3)2(OH2)]2+after which Pt forms covalent bonds with the purine bases at position N7 with the formation magnievykh cross-links (Huifang Huang, Leiming Zhu, Brian R. Reid, Gary P. Drobny, Paul B. Hopkins. Solution Structure of a Cisplatin-Induced DNA Interstrand Cross-Link. Science 1995: 270. 1842-1845). For comparison, carboplatin and oxaliplatin contain CYCLOBUTANE-1,1-dicarboxylate and oxalate, respectively, as leaving groups that chelate platinum more strongly, thereby giving greater stability to the complex leaving group-Pt, and as a result exhibit fewer side effects than cisplatin, but also lower efficiency than cisplatin (Richard J. Knox, Frank Friedlos, David A. Lydall and John J. Roberts Mechanism of Cytotoxicity of Anticancer Platinum Drugs: Evidence That cis-Diamminedichloroplatinum(II) and c?,s-Diammine-(1,1-cyclobutanedicarboxylato) platinum(II) Differ Only in the Kinetics of Their Interaction with DNA. Cancer Research 46, 1972-1979, April 1, 1986; and Ronald S. Go, Alex A. Adjei. Review of the Comparative Pharmacology and Clinical Activity of Cisplatin and Carboplatin. Journal of Clinical Oncology, Vol 17, Issue 1 (January), 1999: 409). The authors of the invention, the polymer chose the 40-dimensional poly(isobutylene-alt-maleic acid) (PIMA or PMA), as each monomer has dicarboxylate group, capable of razvivat complex with cisplatin(Oh)2, thus making it possible to load molecules of cisplatin. In addition, when the hydrogenation of maleic acid is formed succinic acid, which is a component of the Krebs cycle. Poly(isobutylene-alt-maleic acid) 2 was synthesized from poly(isobutylene-alt-maleic anhydride) 1 by reaction with water in the DMF one stage, as shown in Figure 1. The subsequent conjugation of cisplatin with poly(isobutylene-alt-maleic acid) (PIMA) 2 was achieved by mixing hydrated cisplatin for 48 hours with the formation of PMA-Cisplatin 6. Unconjugated cisplatin was removed by dialysis, and the number of loads were determined using NMR spectroscopy and spectrophotometry. Surprisingly, the process of complex formation resulted in the generation of nanoparticles by self-Assembly process, with the size determined by the number of molecules of cisplatin loaded on the polymer. Measurements using dynamic laser scattering showed that the saturation of all sites of complexation of cisplatin leads to the formation of the gel, while the load 15 cisplatin molecules on the polymer leads to the formation of nanoparticles of 100 nm. This was determined using transmission electron microscopy (data not shown).

Cisplatin is the first-line therapy for the treatment of lung cancer and thus the authors and is gaining investigated the effect of PMA-Cisplatin on cell viability Lewis lung cancer. Treatment with cisplatin and PMA-cisplatin caused the same cell death (Fig.1C). However, PMA also induced death of tumor cells. The inventors have discovered that this problem can be solved by using derivatization PMA. The authors of this invention have derivateservlet polymer of Ethylenediamine in alkaline conditions (Fig.2). Surprisingly, despite the fact that the derivatization is not removed cytotoxicity PMA, the cytotoxicity of the complex PMA-cisplatin increased. This may be due to the fact that this group is less strongly bound compared to rederivation PMA. Moreover, such effect was observed in the case carboplatin, which has a lower rate constant of hydration than cisplatin, and the result is also less cytotoxic. The original RMA-cisplatin may be put firmly compared to PMA-EDA because of the strong chelation two carboxypropyl. In order to make the polymer more biocompatible, the inventors have modified polymer of glucosamine (GA). PMA-GA-cisplatin synthesized starting with PMA (1) through reaction with glucosamine and then with an aqueous solution of cisplatin (Fig.3 and 11). All synthesized polymers carrier was latiniroval in the aqueous phase at a room temperature of 25°C for 2 days with gidratirovannym cisplatin as agent patinirovanija that led to established the Yu conjugates. At different points in time, the inventors took aliquots of fines and expected total load of cisplatin on the polymer. The inventors have observed that the effectiveness of the load was ~60% after 5 hours of complexation, ~80% after 30 hours and 100% after 48 hours of patinirovanija. The total load of drugs was 6 mg/15 mg of polymer. Hydrating cisplatin was achieved by using equimolar amounts of cisplatin and AgNOs in dark conditions within 48 hours All media were divided into fractions according to standard procedures by dialysis and isolated using freeze-drying to remove the spectroscopic characteristics. The use of DBU was possible to synthesize the conjugate glucosamine-PMA, as seen on various peaks of the polymer and sugar in the NMR results, which coincide with the expected results of NMR. However, processing bases, triethylamine or DIPEA failed to produce the expected product, but trace amounts in NMR spectra were key to determining the final functional product.

The formation of the complex of cisplatin with PMA-GA resulted in the self-Assembly of complex nanoparticles. In certain cases, the passage of nanoparticles through the filter of 0.22 micron results in the generation of nanoparticles, which are in sub-100 nm range, which is critical for remodeling of homing particles into the tumor using the EPR effect. Surprisingly, studies of the viability of the cells showed that the derived PMA-GA was free from internal toxicity to cells. On the contrary, it supported the effectiveness of hydrated cisplatin (Fig.4B). Moreover, derivatization RMA poliatilenglikole also contributed to the destruction of the inherent toxicity associated with the RMA. In addition, the goal can be achieved by conjugation of maleic acid in the polymer main chain, which is biocompatible.

Increased efficiency derivatizing chelated polymers compared with the original polymer indicates that monocarboxylate-chelated much easier to release the drug showed better activity than dicarboxylato-chelated (6). The inventors have discovered that polymer monocarboxylate-chelated platinum compounds have a significant advantage over the conjugates, in which the metal is connected via a dicarboxylic acid. Even hydrolytic release of the drug from the carrier in monocarboxylate-chelated derivatizing RMA the conjugates compared to the more slow hydrolytic cleavage dicarboxylato-chelated in the PMA may explain the significant difference in cell death. For dallashistory the inventors incubated conjugate the drug-polymer with a lysate of cells of Lewis lung cancer in the chamber for dialysis and quantify the release of free drug using calorimetric analysis. The authors of the invention has been rapid and sustained release of the active agent (Fig.4A). It should be noted that the same composition dialyzed in water for 48 hours to remove any free cisplatin and the authors of the invention have been 100% efficiency load, assuming that the active agent is released in neutral conditions, but quickly released in the presence of lysate of tumor cells.

Described herein compositions can be prepared in the form of gels and used for delivery with a slow release of bioactive agents in specific locations in the subject. For example, the composition can be used for delivery delayed release of platinum compounds in website location of the tumor. In some embodiments, the composition is used for delivery delayed release of the compounds of platinum after the tumor was removed.

The pharmaceutical composition

To introduce the subject associated with the polymer-platinum compounds can be provided in pharmaceutically acceptable compositions. Data pharmaceutically acceptable compositions include a therapeutically effective amount of one or more platinum compounds described herein together with one or more pharmaceutically acceptable but what italiani (additives) and/or diluents. As described in detail below, the pharmaceutical compositions of the present invention may be specially formulated for administration in solid or liquid form, including those which are adapted for the following modes of administration: (1) oral introduction, for example, the dose (aqueous or non-aqueous solutions or suspensions), lozenges, pills, capsules, pills, tablets (e.g., those that are intended for buccal, sublingual, and systemic absorption), boluses, powders, granules, pastes for application to the tongue; (2) parenteral administration, for example, by subcutaneous, intramuscular, intravenous or epidural injection, for example, a sterile solution or suspension, or sustained-release; (3) topical application, for example, in the form of a cream, ointment or patch, or spray with a slow release, applied to the skin; (4) intravaginal or intrarectal, for example, in the form of a pessary, cream or foam; (5) sublingually; (6) ocular; (7) transdermal; (8) transmucosally; or (9) nasal. In addition, the compounds can be implanted in the patient or injected using drug-delivery systems. See, e.g., Urquhart, et al., Ann. Rev. Pharmacol. Toxicol. 24: 199-236 (1984); Lewis, ed. "Controlled Release of Pesticides and Pharmaceuticals" (Plenum Press, New York, 1981); U. S. Pat. No. 3,773,919; and U. S. Pat. No. 35 3,270,960.

Used C the ect, the term "pharmaceutically acceptable" refers to those compounds, materials, compositions and/or dosage forms which are, within a thorough medical check-UPS, are suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic reactions or other problems or complaints, comparable with an acceptable ratio of benefit/risk.

Used here, the term "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition or carrier, such as a liquid or solid filler, diluent, excipient, additive, facilitating production (e.g., lubricant, talc, magnesium stearate, calcium or zinc or stearic acid), or material to encapsulate the solvent involved in the transfer or movement of the test compound from one organ or body part to another body or body part. Each carrier must be "acceptable", that is, to be compatible with other ingredients of the composition and not to be harmful to the patient. Some examples of materials which can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives such as sodium carboxymethylcellulose, methylcellulose, ethical uloza, microcrystalline cellulose and cellulose acetate; (4) powder tragakant; (5) Maestoso; (6) gelatin; (7) lubricants such as magnesium stearate, sodium lauryl sulfate, and talc; (8) excipients, such as cocoa butter and waxes for suppositories; (9) oils, such as peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) bolioli, such as glycerin, sorbitol, lures and polyethylene glycol (PEG); (12) esters, such as etiloleat and tillaart; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic solution; (18) ringer's solution; (19) ethyl alcohol; (20) pH buffered solutions; (21) polyesters, polycarbonates and/or polyanhydrides; (22) fillers such as polypeptides and amino acids; (23) serum component, such as serum albumin, HDL and LDL; (22)2-C12alcohols, such as ethanol; and (23) other non-toxic compatible substances used in pharmaceutical formulations. Wetting agents, dyes, release agents, covering agents, sweeteners, flavors, fragrances, preservatives and antioxidants can also be represented in the composition. Terms such as "excipient", "carrier", "farmace is almost acceptable carrier" or the like are used herein interchangeably.

Used here, the expression "therapeutically effective amount" means that amount of a compound, material, or composition comprising the compound of the present invention that is effective to obtain the desired therapeutic effect, at least in a subpopulation of cells in the animal, at an acceptable ratio of benefit/risk, applicable to any medical treatment. For example, the number of connections, enter the subject, which is sufficient to obtain a statistically significant, measurable change in at least one symptom of cancer or metastasis.

Determination of therapeutically effective amount is within the competence of experienced specialists in this field. In General, therapeutically effective amount may vary depending on the history of the subject, age, condition, sex, and severity and type of health condition of the subject, and other pharmaceutically active agents.

Used herein, the term "introduction" refers to the location of the composition into a subject by a method or regime, resulting in at least partial localization of the composition in the desired location to obtain the desired effect. The compound or composition, described herein, can be administered by any appropriate mode, known in the Noi area, including, but without limitation, oral or parenteral modes, including intravenous, intramuscular, subcutaneous, transdermal, air (aerosol), pulmonary, nasal, rectal and local (including buccal and sublingual) administration.

Examples of modes of introduction include, but without limitation, injection, infusion, instillation, inhalation or ingestion. "Injection" includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intraventricular, intracapsular, intraorbital, intracardially, intradermal, intraperitoneally, transtracheal, subcutaneous, intradermal, intra-articular, subcapsular, subarahnoidalno, intraspinally, intracerebrally and epigastric injection and infusion. In preferred embodiments, the compositions are injected by intravenous infusion or injection.

"Treatment", "warning", or "weakening" of the disease or disturbance mean delay or preventing the onset of such diseases or disorders, reversion, partial withdrawal symptoms, improvement, inhibiting, slowing or stopping the progression, aggravate or deterioration progression or severity of a condition associated with such disease or disorder. In one embodiment, at least one of the symptoms for which Alemania or violation is partially removed, at least 5%, at least 10%, at least 20%, at least 30%, at least 40% or at least 50%.

Used herein, the term "subject" means any person or animal. Typically, the animal is a vertebrate animal, such as a Primate, rodent, pet or hunting animal. Primates include chimpanzees, cynomolgus macaques, spider monkey and monkeys, such as rhesus. Rodents include mice, rats, marmots, ferrets, rabbits and hamsters. Home and hunting animals include cows, horses, pigs, deer, bison, Buffalo, types of cats, such as domestic cats, dog species, such as dogs, foxes, wolves, varieties of birds, such as chickens, EMUs, ostriches, and fish such as trout, catfish and salmon. The patient or subject includes any subset of the above, for example, all of the above, but except for one or more groups or species such as human, non-human primates or rodents. In certain embodiments the subject is a mammal such as a Primate, such as a person. The terms "patient" and "subject" are used here interchangeably.

Preferably, the subject was a mammal. The mammal may be human, nonhuman Primate, mouse, rat, dog, cat, horse is whether the cow, but is not limited to these examples. Mammals other than humans, can be preferably used as subjects that represent animal models of disorders associated with inflammation.

In addition, the methods described herein may be used to treat domesticated animals and/or Pets. The subject may be male or female. The subject may be a subject who was previously diagnosed with or identified as suffering from or having disturbance, cancer or metastasis, but not yet subjected to therapy.

Used here, the term "cancer" includes, but without limitation, solid tumors and tumors of the hematopoietic system. The term "cancer" refers to diseases of the skin, tissues, organs, bones, cartilage, and blood vessels. The term "cancer" also includes primary and metastatic cancer. Examples of cancers that can be treated using compounds of the invention include, but without limitation, cancer, including carcinoma of the bladder, breast, colon, kidney, lung, ovary, pancreas, stomach, cervix, thyroid and skin, including squamous cell carcinoma; hematopoietic tumors of lymphoid origin, including, but without limitation, leukemia, acute lymphocytic leukemia, acute lymphoblastic lake is h, B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, nehodgkinski lymphoma, hairy cell leukemia and Burkitt's lymphoma; hematopoietic tumors of myeloid origin, including, but without limitation, acute and chronic myeloid leukemia, and promyelocytic leukemia; tumors of mesenchymal origin, including, but without limitation, fibrosarcoma, rhabdomyosarcoma, and osteosarcoma; other tumors, including melanoma, seminoma, teratocarcinoma, neuroblastoma and glioma; tumors of the Central and peripheral nervous system, including, but without limitation, astrocytoma, neuroblastoma, glioma and neuromas; and other tumors, including, but without limitation, pigmentary xeroderma, keratoakantoma, follicular thyroid cancer and teratocarcinoma. Compounds of the invention are effective for treating patients who have previously undergone, and are not subjected to anti-cancer therapy. Without a doubt, the methods and compositions of this invention can be used as anticancer therapy first and second line.

Compounds of the invention are also effective in combination with known anti-cancer therapies, including irradiation. The methods of the invention are particularly effective in combination with anti-cancer therapies, which include the introduction is the second drug, the current in the other phase of the cell cycle, such as S-phase, non-epothilone Formula (Ia) or (Ib), which manifest their effect in G2-M phase.

Definition

Unless otherwise stated or implied by context, the following terms and expressions shall have the meanings provided below. Unless specifically provided otherwise or clear from context, the terms and expressions below are not exclusive of value, which term or expression has in the field to which it belongs. The definitions are presented to facilitate the description of the individual cases and do not allow the limitation of the invention described in the claims, as the scope of the invention is limited only by the formula. In addition, unless the context requires otherwise, words in the singular will include the plural, and the terms in the plural will include the singular.

Used here, the term "including" or "includes" is used in reference to compositions, methods, and respective component(s), which are essential to the invention, yet open to the inclusion of unspecified elements, which are essential or not.

Non-working examples, or in cases where otherwise indicated, all numbers expressing quantities of ingredients or reaction conditions used in the estuaries and the e l e C it should be understood as modified in all instances by the term "about". The term "approximately", when used in respect of interest may indicate mean ±1%.

A term denoting the singular "a", "an" and "the" include the plural, unless the context clearly indicated otherwise. Similarly, the word "or" implies the inclusion of "and", unless the context clearly indicated otherwise.

Below is a description of suitable methods and materials, although the practice or testing of this disclosure can be applied to the methods and materials similar or equivalent to those described here. The term "includes" means "includes". The abbreviation "e.g." comes from the Latin for "for example" and is used here to indicate a non-limiting example. Thus, the abbreviation "e.g." is synonymous with the term "for example".

The term "alkyl" refers to saturated non-aromatic hydrocarbon chains that may be a straight or branched chain, containing the indicated number of carbon atoms (these include without limitation methyl, ethyl, propyl, isopropyl, butyl, 2-methyl-ethyl, t-butyl, allyl or propargyl), which can be optional included N, O or S. for Example, C1-C6indicates that the group may contain from 1 to 6 (inclusive) carbon atoms.

The terminology is "alkenyl" refers to alkyl, which contains at least one double bond. Examples alkenyl groups include, for example, but without limitation, ethynyl, propenyl, butenyl, 1-methyl-2-butene-1-yl and the like.

The term "quinil" refers to an alkyl that contains at least one triple bond.

The term "aryl" refers to monocyclic, bicyclic or tricyclic aromatic ring system, in which 0, 1, 2, 3, or 4 atoms of each ring may be substituted by the Deputy. Examples of aryl groups include, but are not limited to, benzyl, phenyl, naphthyl, anthracene, azulene, feranil, indanyl, indenyl, naphthyl, phenyl, tetrahydronaphthyl and the like.

The term "cilil" or "cycloalkyl" refers to saturated and partially unsaturated cyclic hydrocarbon groups containing 3 to 12 carbon atoms, for example from 3 to 8 carbon atoms and, for example, from 3 to 6 carbon atoms, with cycloalkyl group optionally may be optional substituted. Examples cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclooctyl and the like.

The term "heteroaryl" refers to an aromatic 5-8 membered monocyclic, 8-12 membered bicyclic, or 11-14 membered tricyclic ring system containing 1-3 heteroatoms if monocyclic, 1-6 heteroatoms if bicyclic, or 1-9 heteroatoms if tricyclic system, these heteroatoms are selected from O, N or S (for example, carbon atoms and 1-3, 1-6, or 1-9 heteroatoms O, N, or S if monocyclic, bicyclic or tricyclic systems, respectively), with 0, 1, 2, 3, or 4 atoms of each ring may be substituted by the Deputy. Examples of heteroaryl groups include, but without limitation, pyridyl, furyl or furanyl, imidazolyl, benzimidazolyl, pyrimidinyl, thiophenyl or thienyl, pyridazinyl, pyrazinyl, chinoline, indolyl, thiazolyl, naphthyridines and the like.

The term "heterocyclyl" refers to a nonaromatic 5-8 membered monocyclic, 8-12 membered bicyclic, or 11-14 membered tricyclic ring system containing 1-3 heteroatoms if monocyclic, 1-6 heteroatoms if bicyclic, or 1-9 heteroatoms if tricyclic system, these heteroatoms are selected from O, N or S (for example, carbon atoms and 1-3, 1-6, or 1-9 heteroatoms O, N, or S if monocyclic, bicyclic, or tricyclic, respectively), with 0, 1, 2 or 3 atoms of each ring may be substituted by the Deputy. Examples heterocyclyl groups include, but without limitation, piperazinil, pyrrolidinyl, dioxane, morpholinyl, Tetra is hydrofuran and the like.

The term "optional substituted" means that a certain group or a fragment, such as an alkyl group, Alchemilla group and the like, is unsubstituted or substituted by one or more substituents (typically 1-4 substituents) independently selected from the group of substituents listed below in the definition of "Deputy" or stated otherwise.

The term "substituents" refers to a group "substituted" on an alkyl, alkenylphenol, alkenylphenol, cycloalkyl, aryl, heterocyclyl or heteroaryl group at any atom of that group. Suitable substituents include, without limitation, halogen, hydroxy, oxo, nitro, haloalkyl, alkyl, alkenyl, quinil, alkaryl, aryl, aralkyl, alkoxy, aryloxy, amino, acylamino, alkylsulphonyl, arylcarbamoyl, aminoalkyl, alkoxycarbonyl, carboxy, hydroxyalkyl, alkanesulfonyl, arenesulfonyl, alkanesulfonyl, arenesulfonyl, aralkylated, alkylsulphonyl, acyloxy, cyano or ureido. In some cases, two Deputy together with the carbon atoms to which they are attached, may form a ring.

Used herein, the term "polymer" refers to the product of the polymerization reaction, and includes homopolymers, copolymers, terpolymers, terpolymer, etc., the Term "polymer" also includes random polymers, block polymers, graft polymers, the SOP is the materials, block copolymers and graft copolymers. Used here, the term "copolymer" refers to polymers formed by the polymerization reaction of at least two different monomers.

Used herein, the term "main chain of the copolymer" refers to the portion of the polymer, which is a continuous chain that includes the connection formed between the monomers during polymerization. The composition of the main chain of the copolymer can be described from the point of view of the identity of the monomers from which it is formed, without taking into account the composition of branches or side chains of the main polymer chain. The term "side chain" refers to the parts of the monomer which after polymerization to form the lengthening of the main chain of the copolymer.

Used here, the term "biocompatible" refers to a material that is able to interact with the biological system without causing cytotoxicity, unwanted protein or modification of nucleic acid, or activation of unwanted immune reactions. Also "biocompatibility" includes mainly the lack of interaction with recognizing proteins, for example, antibodies of natural origin, cellular proteins, cells and other components of biological systems.

Used here ether side chains mean side chains of the formula-R'”C(O)-OREwhere RE the independent is on represents a C 1-C6alkyl, C1-C6alkenyl, C1-C6quinil, cilil, heterocyclyl, aryl or heteroaryl, each of which may be optional substituted; and R'” represents a bond or C1-C6alkylen, with alkylene may include one or more double or triple links and/or the main chain alkylene may be interrupted O, S, S(O), NH or S(O). Preferably, R'” was a link.

Used here amide side chains mean side chains of the formula R”C(O)-N(RA), where RA independently represents H, C1-C6alkyl, C1-C6alkenyl, C1-C6quinil, cilil, heterocyclyl, aryl, heteroaryl, saccharide, disaccharide or trisaccharide, each of which may be optional substituted; and R" is a bond or C1-C6alkylen, with alkylene may contain one or more double or triple links and/or the main chain alkylene may be interrupted O, S, S(O), NH or S(O). Preferably, R” was a link.

Used here, a chain carboxylic acids means the side chain of the formula R”C(O)OH, where R”” is a bond or C1-C6alkylen, with alkylene may contain one or more double or triple links, and/or the main chain alkylene may be interrupted O, S, S(O), NH or S(O). Preferably, h is Oba R" was a link.

Some non-exhaustive examples of biocompatible polymers include polyamides, polycarbonates, polyalkylene, polyalkylene glycols, polyalkylene oxides, polyalkylene terephthalate, polyvinyl alcohols, polyvinylidene, polyvinyl esters, polyvinylchloride, polyvinylpyrrolidone, polyglycolides, polysiloxanes, polyurethanes and their copolymers, alkylaryl, hydroxyethylcellulose, cellulose ethers, esters of cellulose, nitrocellulose, acrylic polymers and methacrylic esters, methyl cellulose, ethylcellulose, hydroxypropylcellulose, hypromellose, hydroxyethylmethylcellulose, cellulose acetate, cellulose propionate, acetylbutyrate cellulose acetate phthalate cellulose, carboximetilzellulozu, triacetate cellulose, sodium salt of sulfate cellulose, poly(methyl methacrylate), poly(ethyl methacrylate), poly(butylmethacrylate), poly(isobutyronitrile), poly(hexidecimal), poly(isabellemadelinet), poly(laurenmarie), poly(fenilsalicilat), poly(methacrylate), poly(isopropylate), poly(isobutyryl), poly(octadecanoyl), polyethylene, polypropylene, poly(ethylene glycol), poly(ethylene oxide), poly(ethyleneterephthalate), poly(vinyl alcohol), poly(vinyl acetate), polyvinyl chloride, polystyrene, Polygalaceae acids, casein, gelatin, gluten, polyanhydride, polyacryla the traveler acid, alginate, chitosan, any of their copolymers, and combinations thereof. Moreover, biocompatible polymers and copolymers that have been modified for the desired enzymatic cleavage or changed when exposed to light, ultrasound, radiation, changes in temperature, pH, osmotic properties, the concentration of the liquid phase or solvent, is also included in the present invention.

The present invention can be determined using any of the following numbered paragraphs:

1. Biocompatible conjugated polymer nanoparticles including: the main chain of the copolymer;

many of the side chains covalently associated with the specified main chain; and

many platinum compounds, dissociatio associated with these side chains.

2. The nanoparticles under item 1, characterized in that the specified set of platinum compounds is selected from compounds of Pt(II) compounds of Pt(IV) and any combination thereof.

3. The nanoparticles under item 1 or 2, characterized in that at least one of a specified set of platinum compounds is linked to the side chain, at least one coordination bond.

4. The nanoparticles under item 3, characterized in that the coordination bond is between the oxygen of the side chains and the platinum atom of the platinum compounds.

5. The nanoparticles under item 4, the best is the rpm die, that specified by the carbonyl oxygen is oxygen.

6. The nanoparticles under item 4, characterized in that said oxygen is the amide oxygen.

7. The nanoparticles according to any one of paragraphs.1-6, characterized in that the polymer contains a monomer of maleic acid.

8. The nanoparticles under item 7, characterized in that at least one carboxylic acid is maleic acid is derivational in amide.

9. The nanoparticles according to any one of paragraphs.1-8, characterized in that the copolymer is a poly(isobutylene-alt-maleic acid) (PIMA).

10. The nanoparticles according to any one of paragraphs.1-9, characterized in that the copolymer contains from 2 to 100 monomer units.

11. The nanoparticles according to any one of paragraphs.1-9, characterized in that the copolymer contains from 25 to 50 Monomeric units.

12. The nanoparticles according to any one of paragraphs.1-11, characterized in that the said side chains are selected from the group consisting of polymers of monosaccharides, dicarboxylic acids, and combinations thereof.

13. The nanoparticles according to any one of paragraphs.1-12, characterized in that the said side chains are polyethylene glycol (PEG).

14. Of nanoparticles on p. 13, characterized in that the said PEG side chains have a molecular weight of from 100 to 5000 daltons.

15. Of nanoparticles on p. 13, characterized in that the said PEG side chains have the forefront of the lar weight of from 1000 to 3000 daltons.

16. Of nanoparticles on p. 13, characterized in that the said PEG side chains have a molecular weight of about 2000 daltons.

17. The nanoparticles according to any one of paragraphs.1-12, characterized in that these side chains are monosaccharides.

18. The nanoparticles under item 17, characterized in that these monosaccharides is glucosamine.

19. The nanoparticles according to any one of paragraphs.1-18, characterized in that said platinum compound is a compound of Pt(II) selected from the group consisting of cisplatin, oxaliplatin, carboplatin, paraplatin, satraplatin and their combinations.

20. The nanoparticles under item 19, characterized in that said platinum compound (II) is cisplatin.

21. The nanoparticles under item 19, characterized in that said platinum compound is oxaliplatin.

22. The nanoparticles according to any one of paragraphs.1-21, characterized in that the number of side chains is from 50% to 100% of the number of monomer units specified the main polymer chain.

23. The nanoparticles according to any one of paragraphs.1-22, characterized in that the number of the side chains of more than 90% of the number of monomer units specified the main polymer chain.

24. The nanoparticles according to any one of paragraphs.1-23, characterized in that the number of these platinum compounds is from 10% to 100% of the number of monomer units specified the main polymer chain.

25. Nanoc stica according to any one of paragraphs.1-24, characterized in that the number of these platinum compounds ranges from 25% to 75% of the number of monomer units specified the main polymer chain.

26. The nanoparticles according to any one of paragraphs.1-25, characterized in that the said side chains include dicarboxylic acid.

27. The nanoparticles under item 26, characterized in that the said dicarboxylic acids have the formula HOOC-R-COOH, where R represents a C1-C6alkyl, C1-C6alkenyl or2-C6quinil.

28. The nanoparticles under item 27, wherein the specified dicarboxylic acid is maleic acid.

29. Biocompatible conjugated polymer nanoparticles including:

the main chain of poly(isobutylene-alt-maleic acid), characterized in that the main chain contains from 25 to 50 Monomeric units;

many of the side chains of PEG covalently associated with the specified main chain, the PEG side chains have a molecular weight of from 1000 to 3000 daltons, and a specified number of PEG side chains is from 50% to 100% of the number of monomer units specified the main polymer chain; and

many of the side groups of cisplatin, dissociatio associated with the specified main chain, the number of these side groups of cisplatin is from 25% to 75% of the number of monomer units pointed to by the th of the main polymer chain.

30. Biocompatible conjugated polymer nanoparticles including:

the main chain of poly(isobutylene-alt-maleic acid), characterized in that the main chain consists of 40 monomers;

many of the side chains of PEG covalently associated with the specified main chain, the PEG side chains have a molecular weight of 2000 daltons, and a specified number of PEG side chains of more than 90% of monomer units of the specified main polymer chain; and

many of the side groups of cisplatin, dissociatio associated with the specified main chain, the number of these side groups of cisplatin is from 25% to 75% of the number of monomer units specified the main polymer chain.

31. Biocompatible conjugated polymer nanoparticles including:

the main chain of poly(isobutylene-alt-maleic acid), characterized in that the main chain contains from 25 to 50 monomers;

many glucosamine side chains covalently associated with the specified main chain, and the number of these glucosamine side chains is from 50% to 100% of monomer units of the specified main polymer chain; and

many of the side groups of cisplatin, dissociatio associated with the specified main chain, the number of these side groups of cisplatin composition is yet from 25% to 75% of the number of monomer units specified the main polymer chain.

32. Biocompatible conjugated polymer nanoparticles including:

the main chain of poly(isobutylene-alt-maleic acid), characterized in that the main chain contains from 25 to 50 monomers;

many glucosamine side chains covalently associated with the specified main chain, and the number of these glucosamine side chains of more than 90% of monomer units of the specified main polymer chain; and

many of the side groups of cisplatin, dissociatio associated with the specified main chain, the number of these side groups of cisplatin is from 25% to 75% of the number of monomer units specified the main polymer chain.

33. Conjugated nanoparticle complex of carboxylic acid is a compound of platinum, including:

the complex carboxylic acid-platinum compound; and

many lipid-polymer chains, while the carboxylic portion of the specified complex carboxylic acid-platinum compound is covalently associated with these lipid-polymer chains.

34. The nanoparticles under item 33, wherein the carboxylic acid is maleic acid.

35. The nanoparticles according to any one of paragraphs.33-34, wherein the polymer is PEG.

36. The nanoparticles according to any one of paragraphs.33-35, wherein the platinum compound is of the group of Pt(II), selected from the group consisting of cisplatin, oxaliplatin, carboplatin, paraplatin, satraplatin and their combinations.

37. Of nanoparticles on p. 36, characterized in that the compound Pt(II) is cisplatin.

38. The nanoparticles according to any one of paragraphs.33 to 37, characterized in that the load of platinum compounds is from 1% to 30%.

39. The nanoparticles according to any one of paragraphs.33 to 38, characterized in that the load of platinum compounds is from 1% to 6%.

40. Vesicular, micellar or liposomal connection involving many nanoparticles according to any one of paragraphs.33-39.

41. Dicarbonyl-lipid compound having the structure

42. Vesicle, micelle, liposomal or nanomedicine, including dicarbonyl-lipid connection on p. 41 and a platinum compound, wherein the platinum compound is dissociatio associated with the connection on p. 41.

43. The nanoparticles according to p. 42, wherein the platinum compound is selected from compounds of Pt(II) compounds of Pt(IV) and any combinations thereof.

44. Of nanoparticles on p. 43, characterized in that said platinum compound is compounds of Pt(II) selected from the group consisting of cisplatin, oxaliplatin, carboplatin, paraplatin, satraplatin and their combinations.

45. Of nanoparticles on p. 43, characterized in that said connection PLA is ins (II) is cisplatin.

46. Of nanoparticles on p. 43, characterized in that said platinum compound is oxaliplatin.

47. Nanomedicine comprising a biocompatible polymer, wherein the polymer includes at least one monomer having the formula-CH(CO2H)-R-CH(C(O)R')-, where R is a bond, C1-C6alkylene, with alkylene may contain one or more double or triple links; and R' is a substituted nitrogen atom. Preferably, R was a connection.

48. Of nanoparticles on p. 47, characterized in that the polymer contains from 2 to 100 monomer units having the formula CH(CO2H)-R-CH(C(O)R')-.

49. The nanoparticles according to any one of paragraphs.47-48, characterized in that the polymer contains from 25 to 50 Monomeric units having the formula CH(CO2H)-R-CH(C(O)R')-.

50. The nanoparticles according to any one of paragraphs.47-49, wherein R' representsor-NH(CH2CH2O)mCH3where m is 1-150.

51. The nanoparticles according to any one of paragraphs.47-50, additionally comprising bioactive agent.

52. Pharmaceutical composition including:

the nanoparticles or the connection on p. 1-51; and a pharmaceutically acceptable carrier.

53. A method of treating cancer or metastasis, including:

introduction to a subject in need, an effective amount of a composition according to any one of paragraphs.1-52.

54. The method according to p. 53, characterized in that Chirac or metastasis is selected from the group consisting of tumors susceptible or resistant to platinum.

55. The method according to p. 54, characterized in that said cancer or metastasis is selected from the group consisting of breast cancer, head and necks, ovarian, testicular, pancreatic, oral esophagus, gastrointestinal cancer, liver, bladder, lung, melanoma, skin cancer, sarcoma, blood cancer, brain tumors, glioblastomas, tumors of neuroectodermal origin, and any combinations thereof.

56. Way slow release of platinum compounds in a certain place for a subject, comprising: providing the location of the composition according to any one of paragraphs.1-52.

57. The method according to p. 56, wherein the composition is in gel form.

58. The method according to any of paragraphs.56-57, wherein the location is a tumor.

59. The method according to p. 58, characterized in that the tumor was removed to ensure the composition.

In unspecified cases medium-sized specialists in this area should be clear that any of the different variants described and illustrated herein may be further modified to include characteristics shown in other embodiments disclosed here.

The implementation of the invention

The following examples illustrate some of the options and aspects of the invention. Specialists in this region, the STI will be obvious, that various changes, additions, substitutions and the like can be made without departure from the essence or scope of the invention, and such modifications and variations are included in the scope of the invention as defined in the following formula. The following examples do not limit the invention in any way.

EXAMPLES

Materials and methods

Reagent CellTiter 96 AQueous One Solution Cell Proliferation Assay [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxy)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt; MTS] was obtained from Promega (Madison, WI). All polymer solutions were subjected to dialysis using dialysis tubing cellulose membrane Spectra/Por 4 and Spectra/Por 6 (wet pipe), with cut points mass-average molecular weight of 1000 and 3500, respectively. The process performed against several parties mixed with deionized H2O. Commercially purchased (Sigma, Fluka AG, Aldrich Chemie GmbH) chemical purity higher technical used in the form in which they were received. These include N,N-dimethylformamide (DMF), poly(isobutylene-alt-maleic anhydride), glucosamine. HCl, mPEG2000NH2, diaza(1,3)bicyclo[5.4.0]undecane (DBU), triethylamine.1H NMR and13With NMR were measured at 300 and 400 MHz, respectively, using a spectrometer Varion-300 or Bruker-400. Chemical shifts1H NMR represented as 8 values in parts per million is (M. D.) on tetramethylsilane (0.0 M. D.) or deuterium oxide (4.8 memorial plaques). Data are presented as follows: chemical shift, multipletness (s=singlet, d=doublet, t=triplet, q=Quartet, m=multiplet, b=broad), constant interaction (Hz) and integration. The chemical shifts of the carbon-13 are presented in memorial plaques relative to CDCl3 (76.9 M. D.) or relatively DMSOd6(39.5 memorial plaques). Chemical shifts195Pt NMR is presented as 5 in memorial plaques relative to Na2PtCl6(0.0 m D.). In some experiments1H NMR and13With NMR were measured at 500 and 125 MHz, respectively, using a spectrometer Varion 500 or Bruker-400.

The original materials were obezvozhivani azeotropic distillation until the beginning of the reaction as necessary, and all air and/or moisture-sensitive reactions were performed in dried in the flame of the burner and/or a drying Cabinet glass container in a dry nitrogen atmosphere using standard precautions to exclude moisture.

Cell culture and analysis of viability of cells

Cell lines of Lewis lung carcinoma (LLC) and cell line breast cancer (T) were purchased from American Type Culture Collection (ATCC, Rockville, MD, USA). Cells of Lewis lung carcinoma were cultured in modified according to the method of Dulbecco environment, Needle, supplemented with 10% FBS, 50 units/ml penicillin and 50 units/ml streptomycin. Cells T were cultured in RPMI medium, supplemented with 10% FBS, 50 is Diniz/ml penicillin and 50 units/ml streptomycin. Treated with trypsin cultured LLC cells and T washed twice by PBS and seeded in 96-well tablets with a flat bottom at a density of 2×103cells in 100 µl medium. Different concentrations of the conjugates were tested with the three-fold repetition in the same 96-well tablets for each experiment. The environment was kept as negative control and CDDP as a positive control. The tablets are then incubated for 48 h in an atmosphere of 5% CO2at 37°C. Cells were washed and incubated with 100 μl of medium without phenol red (without FBS) containing 20 μl of CellTiter 96 Aqueous One Solution reagent (Promega, WI, USA). This analysis using [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxy)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt] (MTS) is a colorimetric method for determining the number of viable cells in the tests on the proliferation or cytotoxicity. After 2 h incubation in an atmosphere of 5% CO2at 37°C, the absorption in each well was recorded at 490 nm using a tablet reader (VERSA max, Molecular Devices, Sunnyvale, CA, USA).

The acquisition reflects the number of surviving cells. A control sample was subtracted from all data, and the results were analyzed using the software Origin software (OriginLab Corporation, Northampton, USA). The average value of the data acquisitions in three replications for each test dose was divided by the average value of the La untreated control cells. The logarithm of the relations used for plotting as a function of the given dose, i.e. Y=(average value of absorption of tested samples - background)/(average value of the absorbance of untreated samples - background) depending on X=test dose.

Measurement of particle size

The high-resolution images were obtained on a high-contrast digital THE JEOL 2011. Samples were prepared coated carbon copper grids, 300 mesh (Electron Microscopy Sciences) by adding drops of aqueous solutions of nanoparticles at various concentrations and left to air dry. The distribution of the nanoparticles size was studied using the method of dynamic light scattering (DLS), which was performed at 25°C on DLS system (Malvem NanoZetasizer) equipped with a He-Ne laser.

The study of the kinetics of physico-chemical release PIMA-GA-CDDP suspended in 1 ml of lysate hypoxic cells cell line LLC and tightly closed in a dialysis bag (MWCO~1000 Da). The dialysis bag was incubated in 1 ml PBS buffer at room temperature with vigorous shaking. Aliquot 10 µl were taken from the incubation medium at predefined intervals of time, was treated with 90 μl of a solution of 1,2-phenylenediamine (1.2 mg in 1 ml DMF) and incubated for 3 h at 100°C. Freed Pt(IV) quantitatively evaluated using UV-VIS spectroscopy at characteristices the th wavelength X=704 nm of Pt(IV) complex-1,2-phenylenediamine. After selection of each aliquot of the incubation medium were added to 10 μl of fresh PBS.

Alternatively, the concentrated conjugate PIMA-GA-cisplatin resuspendable in 100 ál of bidistilled water at pH is brought to 8.5 or 5.5 using IN sodium hydroxide or IN nitric acid, and was transferred into a dialysis tube (MWCO: 1000 KD, Spectrapor). The dialysis tube was placed in a tube containing magnetic beaker and 2 ml of phosphate-saline buffer solutions with different pH. The release of cisplatin was studied by vigorous mixing of the dialysis bag at 300 rpm using a stirrer IKA at 25°C. From the external solution dialysis membrane bag, taking aliquots of 10 µl of the pre-defined intervals of time, and then was subjected to the next reaction is the formation of an active complex using UV-Vis by adding 100 μl of ortho-phenyldiamine (1.2 mg/ml in DMF) and heating the resulting solution for 3 hours 10 ál of freshly prepared solution was added back to the external solution dialysis membrane bag to maintain the same volume. The amount of released drug was estimated using UV-spectrophotometer (Shimadzu UV 2450) at 706 nm.

FACS analysis of apoptosis

Cells were grown in 6-hole tablets were incubated in the presence of nanoparticles of cisplatin or free cisplatin at 37°C for 24 the. After 24 h the cells were washed by PBS and collected at 0°C. Then cells were treated with conjugate annexation V-Alexa Fluor 488 (Molecular Probes, Invitrogen) and incubated in the dark at room temperature for 15 minutes the Cells were then washed by PBS, and incubated with a solution of propecia iodide (PI) (50 g/ml; Sigma) containing RNase (1 mg/ml; Sigma). Cell suspension is then transferred into FACS tubes and analyzed for staining on the Annexation V/PI flow cytometer BD FACS Calibur. Data were analyzed using CellQuestPro software (BD Biosciences).

The study of cellular uptake

Cells LLC and T were sown on top of the glass in a 24-hole tablets, 50,000 cells per well. After reaching 70% of the population of cells was treated fluoresceinisothiocyanate (PITC)-conjugated nanoparticles of cisplatin during different periods of time, 30 min, 2 h, 6 h, 12 h and 24 h, respectively. To study colocalization the indicated time points the cells were washed by PBS, and incubated with Lysotracker Red (Molecular Probes) at 37°C for 30 min for internalization. Cells then were fixed in 4% paraformaldehyde for 20 min at room temperature, then washed twice by PBS and placed on glass slides using reagent Prolong Gold Antifade Reagent (Molecular Probes). Images were obtained using a fluorescent microscope Nikon Eclipse TE, has a green and red f is ltraj for FITC and Lysotracker Red, respectively.

In Vivo murine models of tumor LLC lung cancer and breast cancer T Cell lung cancer (LLC) and the breast cancer cells T (3×105) were implanted subcutaneously in the lateral region of 4-week-old mice S/BL6 and BALB/c mice (weight 20 g, Charles River Laboratories, MA), respectively. Drug therapy was started after reaching the tumor volume of 50 mm3. Therapy of the tumor was the introduction of nanoparticles of cisplatin and free cisplatin or oxaliplatin and free oxaliplatin. The compositions were prepared and alidibirov so that 100 μl of nanoparticles of cisplatin and free cisplatin contained 1.25 and 3 mg/kg cisplatin, or 100 μl of nanoparticles oxaliplatin and free oxaliplatin contained 5 and 15 mg/kg oxaliplatin. The introduction is carried out by injection into the tail vein. PBS (100 μl), introduced by injection into the tail vein, was used as control drug therapy. The volume of tumor and body weight was monitored on a daily basis. Animals were slaughtered when the average tumor size exceeding 2000 mm3in the control group. Tumors were removed immediately after slaughter and stored in 10% formalin for further analysis. All transactions with animals were approved by the Harvard institutional IUCAC.

In Vivo murine tumor model of ovarian cancer

Adenocarcinoma of the ovary induced in genetically-created mice with K-rasLSL /PtenfI/fIby intrabursal delivery of adenovirus carrying Cre-recombinases, as described previously. Tumor cells were created for the expression of luciferase after activation adeno-Cre to allow for tumor scintigraphy before and after drug therapy. After development in mice tumors medium or large in size, the mice were divided into four treated groups (control, cisplatin NP 1.25 mg/kg cisplatin NP-3 mg/kg and free cisplatin), all drugs were injected intravenously (i.v.).

Scintigraphy tumors and evaluation of the effectiveness of drug therapy

Scintigraphy tumors in vivo was performed using the imaging system IVIS Lumina II Imaging System. Quantification of bioluminescence were obtained using the software Living Image 3.1 Software (Caliper Life Sciences). Mice received 150 mg/kg of potassium salt of D-luciferase Firefly by intraperitoneally (i.p.) injection before imaging. Five minutes after injection of luciferin animals were anestesiologi in an induction chamber with 2.5% isoflurane. After anesthesia, the mice were placed in a cell to render, where they were kept under General anesthesia by filing izoflurana, and the temperature of their bodies maintained at 37°C. Bioluminescent signal was recorded 15 minutes after the introduction of luciferin during the exposure time of thirty seconds is. Image did the day before treatment (day 0, baseline), in the middle of the loop and the next day after final treatment. Treatment effectiveness was evaluated quantitatively by examining fold increase in bioluminescence signal after processing compared with the reference signal. Statistical analysis of toxicity data were analyzed using one-way analysis of variance ANOVA using the software Prism 5™ software.

The biodistribution of cisplatin

To study the distribution of the mice by intravenous i.v. were injected with nanoparticles of cisplatin and free cisplatin (dose equivalent to 8 mg/kg cisplatin). 24 hours after injection the animals were killed and the dissection was performed to extract the tumor and kidney. In another study, animals were dosed out several times according to the protocols of the study of efficiency and cut in the end of the study, repeated administration of doses. The bodies were then weighed and dissolved in conc. HNO3(approximately 10 ml) by shaking for 24 hours at room temperature and subsequent heating at 100°C for 12 hours. The resulting mixture was added 30% H2O2the solution was stirred for 24 hours at room temperature and then heated for 12 hours to evaporate the liquids. All solid residues were re-dissolved the in 1 ml of water and then measured the amount of platinum by the method of mass spectrometry with inductively coupled plasma (ICP).

Gepatologicheskiy and TUNEL analysis (analysis of apoptosis)

Tissues were fixed in 10% formalin, embedded in paraffin, cut into sections and stained with H&E at Harvard Medical School Core Facility. Paraffin sections of the tumor and kidney were deparaffinization and were processed using TUNEL (Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling) - TMR staining (red) according to the manufacturer's Protocol (In Situ Cell Death Detection Kit, TMR Red, Roche). Images were obtained using a fluorescent microscope Nikon Eclipse TE2000, equipped with a red filter.

Evaluation of toxicity of drug therapy

To assess the toxicity of daily recorded body weight. In addition, at the end of processing of the liver and spleen were removed for recording weight and perform extensive anatomical and pathological studies to assess toxicity to vital organs. Cell apoptosis in vital organs was measured using the TUNEL method. Statistical analysis of toxicity were performed using two-factor analysis of variance ANOVA using the software Prism 5™ software.

Statistical analysis

Data are presented as mean±STD.off. from at least n=3. Statistical analysis was performed using the software GraphPad Prism software (GraphPad, San Diego, CA). Statistical analysis of obtained data was performed using the dispersion anal is for ANOVA with subsequent a posteriori comparisons criterion Newman-Keuls or t-test t-test. p<0.05 was intended to indicate significant differences.

Example 1 - Synthesis of polymer carriers

Poly(isobutylene-alt-maleic acid) PIMA (2)

Poly(isobutylene-alt-maleic anhydride) 1 (1 g) was dissolved in 5 ml of dry DMF in a 10 ml flask with a round bottom, which was added bidistilled water (1 ml), and then the resulting reaction mixture was stirred at 80°C for 48 hours the Solvent was removed in vacuum and low molecular weight impurities were removed by dialysis. The aqueous polymer solution was dialyzed for 3 days in the tubing cellulose membrane Spectra/Por 4 and cutoff mass-average molecular weight of 1000. The colorless solution is then liofilizirovanny to obtain 732 mg of a white polymer, poly(isobutylene-alt-maleic acid) PIMA (2).1H NMR (300 MHz, D2O) δ 3.3-3.5 (m), 2.8 (s), 2.6-2.7 (m), 2.5 (s), 2.2-2.3 (m), 0.8-0.9 (m).

PIMA-EDA (3)

A 10 ml flask with a round bottom equipped with a magnetic stirrer and container with dry nitrogen, were loaded poly(isobutylene-alt-maleic anhydride) PIMA 1 (1 g), dry DMF (5 ml), triethylamine (0.1 ml) and excess of the Ethylenediamine dihydrochloride (1 g). The resulting mixture was stirred at 25°C for 48 hours the Solvent was removed in vacuum and the polymer was purified by removing low molecular weight impurities, such as excess Ethylenediamine, using a dialysis bag with a cutoff molecular weight of 3.5 kDa is 3 days. The polymer solution is then liofilizirovanny to obtain 0.89 g PIMA-EDA (3).1H-NMR (300 MHz, D2O) 5 3.1-3.2 (m), 2.9-3.0 (m), 2.6-2.8 (m), 2.5 (m), 0.8-1.0 (m).

The polymer PIMA-GA (4)

Poly(isobutylene-alt-maleic anhydride) PIMA 1 (0.0064 g, 0.001 mmol) was dissolved in DMF (5 ml) and then added DBU (0.032 ml, dissolved in 1 ml dry DMF, 0.21 mmol) and the mixture was stirred at 25°C for 1 h In this solution was directly added glucosamine (0.046 g, 0.21 mmol). The resulting reaction mixture was allowed to mix at room temperature for 48 h and then besieged by adding bidistilled water (1 ml). The organic solvent is evaporated in vacuum for 12 hours. The obtained pale yellow solid was purified by dialysis for 3 days using a dialysis bag, obtained from Pierce (Thermoscientific) with a cutoff molecular weight of 3.5 kDa to colorless solution. Lyophilization allowed to obtain 104 mg of polymer PIMA-GA white (4).1H-NMR (300 MHz, CDC13) δ 7.54-7.65 (m, 2 H), 7.33-7.45 (m, 2 H), 7.02-7.19 (m, 14 H), 6.93-6.97 (m, 2 H), 6.83-6.89 (m, 2H), 6.55 (s, 2H), 6.15-6.19 (m, 2H), 3.90 (s, 2 H), 3.58 (s, 6 H).1H-NMR (300 MHz, D2O) δ 8.2-8.3 (m), 7.0-7.1 (m), 5.0-5.1 (m), 3.0-3.9 (m), 2.1-2.3 (m), 1.1-1.9 (m), 0.7-1.0 (m).

In another experiment PIMA (0.045 g) was dissolved in DMF (5 ml) and then was added a solution of DBU (0.23 ml) and glucosamine (0.323 g was dissolved in 5 ml dry DMF). The resulting reaction mixture was left premesis is sterile at room temperature for 48 h and then besieged by adding bidistilled water (1 ml). The organic solvent evaporated in vacuum. The obtained pale yellow solid was purified by dialysis for 3 days using a dialysis bag with a nominal cutoff molecular weight of 3.5 kDa. Lyophilization allowed to obtain 104 mg of a slightly yellow polymer PIMA-GA.1H-NMR (300 MHz, D2O) δ 8.2-8.3 (m), 7.07.1 (m), 5.0-5.1 (m), 3.0-3.9 (m), 2.1-2.3 (m), 1.1-1.9 (m), 0.7-1.0 (m).

The polymer PIMA-PEG (5)

Poly(isobutylene-alt-maleic anhydride) PIMA 1 (3 mg, 0.0005 mmol) and DBU (0.0023 ml, 0.015 mmol) was dissolved in dry DMF (10 ml) in a 25 ml flask with a round bottom in the atmosphere N2for 1 h and then added to the PEG-NH2(20 mg, 0.01 mmol), the resulting reaction solution was then heated at 80°C under continuous stirring for 3 days. The reaction mixture was left to cool to room temperature and then added water (1 ml) and continued stirring for 1 h the Solvent was removed in vacuum and unreacted PEG-NH2with MW 2 kDa were removed from the desired polymer by dialysis. Dialysis was carried out for 5 days using a membrane with a cutoff molecular weight of 3.5 kDa, obtained from Pierce (Thermoscientific), with the formation of a colourless solution, which is then liofilizirovanny to obtain 19 mg of a white PIMA-PEG (5).1H-NMR (300 MHz, D20) δ 3.5-3.7 (m), 3.0-3.1 (m), 2.5-2.8 (m), 0.7-1.0 (m).

Example 2 - Synthesis of conjugates

Hydrating the CDDP

CDDP (30 mg) and AgNOa (17 mg) was added in 10 ml bidistilled water. The resulting solution was stirred in the dark at room temperature for 24 hours After completion of the reaction was found to precipitate AgCl. The precipitated AgCl was removed by centrifugation at 10,000 rpm for 10 min. the Supernatant was further purified by passing through a 0.2 μm filter.

PIMA-CDDP (6}

Poly(isobutylene-alt-maleic acid) PIMA 2 (0.006 g, 0.001 mmol) was dissolved in 1 ml of bidistilled water containing CDDP (0.00084 g, 0.0028 mmol), 10 ml flask with a round bottom and then the resulting reaction mixture was stirred at room temperature (25°C) for 48 hours Conjugate PIMA-CDDP (6) then purified by lisalisa in the tube of cellulose membranes Spectra/For 4 and cutoff mass-average molecular weight of 1000. The obtained opaque solution is then liofilizirovanny for white conjugate PIMA-CDDP (6). Conjugate resuspendable for experiments with cell culture.

PIMA-EDA-CDDP (7}

A 10 ml flask with a round bottom loaded polymer PIMA-EDA 3 (0.007 g, 0.001 mmol), which was added CDDP (0.0084 g, 0.0028 mmol), dissolved in bidistilled water (1 ml). The solution is then stirred at room temperature (25°C) for 48 hours Dialysis using a cellulose membrane with a cutoff threshold of molecular weight 1000 and lyophilization allowed to obtain yellowish conjugate PIMA-EDA-CDDP (7).

PIMA-GA-CDDP (8)

A 10 ml flask with a round bottom equipped with a magnetic stirrer, measured PIMA-GA 4 (0.0036 g, 0.0003 mmol), was added 1 ml of bidistilled water containing CDDP (0.001 g, 0.0033 mmol), after which the solution was stirred at room temperature (25°C) for 48 hours Resulting in a conjugate solution PIMA-GA-CDDP (8) was purified by dialysis to remove unattached CDDP with cutoff mass-average molecular weight of 1000 in 2-3 hours. Lyophilization detalizirovannoi solution allowed to obtain slightly yellow conjugate PIMA-GA-CDDP (8).

PIMA-PEG-CDDP (9}

Brush polymer PIMA-PEG 5 (0.019 g, 0.00007 mmol) were placed in a 10 ml flask with a round bottom, mixed with CDDP (0.0002 g, 0.0007 mmol), dissolved in 0.3 ml of bidistilled water. After stirring for 3 days at room temperature (25°C) resulted in an opaque reaction mixture was subjected to dialysis. The solution containing the conjugate PIMA-PEG-CDDP (2), then purified by dialysis tubing cellulose membrane Spectra/For 4 and cutoff mass-average molecular weight of 1000 for 2-3 hours to remove free CDDP. Conjugate PIMA-PEG-CDDP (9) then liofilizirovanny to obtain white solids. Conjugate resuspendable in bidistilled water for experiments with cell culture.

FITC-labeled PIMA-GA-CDDP

Poly(isobutylene-alt-maleic anhydride) PIMA (0.006 g) rest rely in DMF (5 ml) and then was added a solution of DBU (0.0053 ml in DMF) and glucosamine (0.0075 g) was dissolved in 5 ml dry DMF), the mixture was stirred at 25°C for 1 h the reaction mixture was allowed to mix at 25°C for 48 h and then added 0.0022 g FITC-EDA (FITC-EDA was synthesized by mixing fluoresceinisothiocyanate in excess of Ethylenediamine at 25°C for 12 h in DMSO), and the stirring continued for another 12 h, after which the reaction mixture was besieged by adding bidistilled water (1 ml). The organic solvent evaporated in vacuum. The obtained orange solid was purified by dialysis for 3 days using a dialysis bag with a cutoff molecular weight of 3.5 kDa. Lyophilization allowed to obtain fluorescent orange polymer PIMA-GA-FITC. In this FITC-labeled polymer (PIMA-GA-FITC, 0.004 g) was added to 1 ml of bidistilled water containing cisplatin (0.001 g) and then the solution was stirred at room temperature (25°C) for 48 hours Conjugate PIMA-GA-FITC-cisplatin formed in solution, and then purified by dialysis to remove unattached cisplatin with cutoff mass-average molecular weight of 1000. Lyophilization cialisovernight solution was allowed to get orange nanoparticles conjugate FITC-labeled PIMA-GA FITC-cisplatin.

PIMA-Oxaliplatin

Poly(isobutylene-alt-maleic acid) (PIMA) (6 mg) was dissolved in 1 ml of bidistilled water containing oxaliplatin the h-HE (1 mg), in the flask with a round bottom and then the resulting reaction mixture was stirred at room temperature (25°C) for 48 hours Conjugate PIMA-oxaliplatin was further purified by dialysis tubing cellulose membrane Spectra/Por 4 and cutoff mass-average molecular weight of 1000. The obtained opaque solution is then liofilizirovanny to obtain conjugate PIMA-oxaliplatin. Conjugate resuspendable for experiments with cell culture.

PIMA-GA-Oxaliplatin

In PIMA-GA (12 mg), measured in a 10 ml flask with a round bottom equipped with a magnetic stir bar, was added 1 ml of bidistilled water containing oxaliplatin-HE (1 mg), and then the solution was stirred at room temperature (25°C) for 48 hours Conjugate PIMA-GA-oxaliplatin, the resulting solution was purified by dialysis to remove unattached oxaliplatin with cutoff mass-average molecular weight of 1000. Lyophilization detalizirovannoi solution allowed to obtain yellow conjugate PIMA-GA-oxaliplatin.

Example 3 - NMR analysis of the synthesis of polymer PIMA-GA using different bases

Synthesis PIMA-GA using DBU as the base

Glucosamine hydrochloride (360 mg, 1.66 mmol, 200 equiv.) suspended in 5 ml of DMF and was treated with DBU (250 μl, 1.66 mmol, 200 equiv.) at room temperature for 1 h After 1 h, the solution glucosamine/DBU (DMF) EXT is ulali dropwise into a solution of poly(isobutylene-alt-maleic anhydride) (50 mg, 0.008 mmol, 1 EQ.) in 5 ml of DMF and the reaction mixture was stirred for 72 h at room temperature. The reaction mixture was besieged by 3 ml of bidistilled water. Conjugate PIMA-GA was purified by dialysis using a dialysis bag MWCO 2000 within 72 hours Product liofilizirovanny for 48 h to obtain 100 mg of cream-yellow powder. The product was analyzed by spectroscopy1H NMR (300 MHz). Solubility: the product was soluble in water but insoluble in organic solvent, such as acetone, methanol or acetonitrile.1H NMR (300 MHz): 5 (M. D.)=5.2-5.3 (m, 0.14 H, sugar proton), 5.0-5.1 (m, 0.4 H, sugar proton), 3.6-4.0 (m, 13.07 H, sugar proton), 3.25-3.5 (m, 15.48, H, sugar proton), 3.0-3.2 (m, 6.98 And the sugar proton), 2.5-2.6 (m, 6.97 N, PIMA proton), 1.4-1.7 (m, 19.86 N, PIMA proton), 0.7-1.2 (m, at 23.77 N, PIMA proton). The total number of protons sugar: total number of PIMA protons=36.07: 50.6=0.71. The result is in good agreement with the proposed structure, if all residues are derivationally protons of sugar and PIMA protons in the monomer conjugate PIMA-GA.

Synthesis PIMA-GA using diisopropylethylamine (DIPEA) as the base

Glucosamine hydrochloride (179 mg, 0.83 mmol, 100 EQ.) suspended in 2 ml of DMF and was treated with DIPEA (145 μl, 0.83 mmol, 100 EQ.) at room temperature for 1 h After 1 h the reaction mixture was added a solution of poly(isobutylene-alt-maleic of any the reed) (50 mg, 0.008 mmol, 1 EQ.) (dissolved in 3 ml of DMF and was stirred for 24 h at room temperature. The reaction mixture was precipitated with addition of 3 ml of bidistilled water. Conjugate PIMA-GA was purified by dialysis using a dialysis bag MWCO 1000 within 24 hours Product liofilizirovanny for 48 h to obtain 106 mg of a white powder. The product was analyzed by spectroscopy1H NMR (300 MHz). Solubility: the product was soluble in water but insoluble in organic solvent, such as acetone, methanol or acetonitrile.1H NMR (300 MHz): δ (M. D.)=5.2-5.3 (m, 0.4 H, sugar proton), 4.9-5.1 (m, 2.0 H, sugar proton), 3.4-3.6 (m, 21.86 H, sugar proton), 3.2-3.3 (m, 6.16 H, sugar proton), 2.9-3.1 (m, 3.81 H, sugar proton), 2.4-2.7 (broad, 4.39 N, PIMA proton), 2.1-2.4 (broad, 4.54 N, PIMA proton), 1.7-2.0 (broad, 3.13 N, PIMA proton), 1.3-1.5 (broad, 1.58 N, PIMA proton), 1.1-1.2 (m, 24.12 N, PIMA proton), 0.6-0.9 (m, at 27.94 N, PIMA proton). All protons of sugar: total PIMA protons=39.21:61.11=0.64.

Synthesis PIMA-GA using triethylamine as the base

Glucosamine hydrochloride (143 mg, 0.66 mmol, 80 EQ.) suspended in 2 ml of DMF and treated with triethylamine (100 μl, 0.66 mmol, 80 EQ.) at room temperature for 1 h After 1 h the reaction mixture was added poly(isobutylene-alt-maleic anhydride) (50 mg, 0.008 mmol, 1 EQ.) and was stirred for 24 h at room temperature. The reaction mixture is precipitated with addition of 3 ml of bidistilled water. Conjugate PIMA-GA was purified by dialysis using a dialysis bag MWCO 1000 within 24 hours Product liofilizirovanny for 48 h with 100 mg of white powder. The product was analyzed by spectroscopy1H NMR (300 MHz). Solubility: the product was soluble in water but insoluble in organic solvent, such as acetone, methanol or acetonitrile.1H NMR (300 MHz): δ (M. D.)=5.2-5.3 (m, 0.44 H, sugar proton), 4.9-5.1 (m, 1.51 H, sugar proton), 3.7-3.8 (m, 19.01 H, sugar proton), 3.3-3.4 (m, 6.43 H, sugar proton), 3.1-3.2 (m, 11.82, sugar proton), 2.93-2.94 (m, 2.23 N, PIMA proton), 2.6-2.7 (m, 5.84 N, PIMA proton), 2.2-2.5 (broad, 4.91 N, PIMA protons), 1.8-2.1 (broad, 3.83 N, PIMA proton), 1.4-1.6 (broad, 2.52 N, PIMA protons), 1.8-1.2 (m, 18.04 N, PIMA proton), 0.9-1.0 (m, at 23.77 N, PIMA proton). The total number of protons sugar: total number of PIMA protons=34.31:65.7=0.52.

Example 4 - Zawischa time efficiency load PIMA-GA-CDDP

Method: conjugate PIMA-GA (50 mg, 0.004 mmol) was dissolved in 1 ml of bidistilled water and then adding (NH2)2Pt(OH)2(3 ml, 0.057 mmol). The reaction mixture was stirred at room temperature for 48 hours aliquots of 200 µl were taken from the reaction mixture at pre-defined intervals of time (5 h, 31 h and 48 h). The aliquot was filtered through a centrifugal filter device Microcon containing a membrane made of regenerated cellulose, MWCO 3000, to separate conjugate PIMA-GA-CDDP. The polymer was thoroughly washed (200 ál x 2) bidistilled water to remove platinum reagents. The content of platinum in the polymer was determined by the method described previously.

Result: the Change in efficiency load Pt conjugate PIMA-GA was determined by the ability of the conjugation of 1,2-phenylenediamine with Pt, causing an increase in UV-VIS spectrum at a wavelength of λ=706 nm. Neither the polymer nor 1,2-phenyldiamine had no characteristic peak at this wavelength. Change the content of Pt over time in the reaction between PIMA-GA and hydroxy-platinum was observed using UV-VIS spectrum. At predetermined points in time (5 h, 31 h and 48 h) from the reaction mixture were collected aliquots of 200 µl and was determined load Pt conjugate polymer. The graph shows that the load of platinum in the conjugate polymer increases with time from 190 µg/mg (5 h) to 210 µg/mg (31 h) and reaches a maximum 347 µg/mg in 48 hours, It indicates that almost 100% Pt forms a complex with the polymer at this point in time, as the maximum theoretically calculated load is 37.5% unit of the polymer, and achieved load amounted to 34.7% Pt on the polymer.

Example 5 - a Rational optimization of the polymer, based on the dependence of activity on structure

To improve the efficiency of the nanoparticles, the authors of the invention have derivateservlet one beam of each monomer of the polymer BIOS is compatible with glucosamine for education conjugate PIMA-glucosamine (PIMA-GA) (Fig.11B). It turned dicarboxylato connection with Pt in monocarboxylate communication and coordination, which may be easier to releases Pt, as it is less stable than monocarboxylate relationship (Fig.11B).

Analysis by the method of nuclear magnetic resonance (NMR) environment Pt showed that the complexation PIMA-GA and cisplatin at acidic pH (pH 6.5) was generated isomeric state [PIMA-GA-Cisplatin (O->Pt)] (8), characterized monocarboxylate and O->Pt coordination complex, as seen in one peak Pt NMR spectrum at -1611.54 (Fig.11B). Surprisingly, the formation of the complex of cisplatin with PIMA-GA at alkaline pH (pH 8.5) contributes to the formation of isomeric complex PIMA-GA-Cisplatin (N->Pt) (10), in which the complexation of Pt occurs through monocarboxylate and more stable N->Pt coordination bond, characterized by a single peak at -2210 (Fig.11B). Surprisingly, the existence of these two pH-dependent States have allowed the inventors to analyze the influence of environment, Pt, especially the leaving groups on the biological efficiency.

Complexation of cisplatin with polymer PIMA-glucosamine (PIMA-GA) at a ratio of 15:1 has caused the self-Assembly of the nanoparticles in the desired narrow size range that makes 80-150 nm, which was confirmed by electron microscopy, high-resolution (data not shown)and DLS (Fig.12A). In addition, the inventors have reached a load of 175±5 µg/mg polymer (Fig.12V), which was considerably higher than the load that is possible when using traditional nanosustain (Avgoustakis K, Beletsi A, Panagi Z, Klepetsanis P, Karydas AG, Ithakissios DS. PLGA-mPEG nanoparticles of cisplatin: in vitro nanoparticle degradation, in vitro drug release and in vivo drug residence in blood properties. J Control Release. 2002 Feb 19;79(1-3):123-35).

Example 6 Characterization of the absorption and efficacy of nanoparticles in vitro

Tagging polymer with fluorescein (Fig.15) allowed us to track over time the uptake of nanoparticles by cells that were together in the state of a fluorescent dye LysoTracker-Red for marking endolysosomal compartments. The rapid uptake of nanoparticles was observed in LLC cells within 15 min of treatment with internalization in endolysosomal compartment, as seen in colocalization FITC-labeled nanoparticles and dye LysoTracker-Red (data not shown). In contrast, uptake by cells T was slow with internalization in endolysosomal compartment, expressed only 2 hours after incubation. During the 12-hour period of the fluorescent signals from lysosomal compartment and FITC-conjugated separated, which involves cytosolic distribution of the polymer after processing within a complementary mechanism (data not shown).

To test the effectiveness of the nanoparticles PIMA-GA-cisplatin in vitro, the inventors conducted an is Liz viability of cells using cell lines of Lewis lung carcinoma (LLC) and breast cancer T. Cell viability was quantitatively assessed using the MTS analysis at 48 hours after incubation. Surprisingly, LLC cells (Fig.13A) were more sensitive to cisplatin nanoparticles than breast cancer cells T (Fig.13B). Surprisingly, the nanoparticles PIMA-GA-cisplatin (O->Pt) (8) showed a significant loss of LLC cells with IC50 values of (4.25±0.16 μm), similar (P>0.05) this drug called cisplatin (IC50=3.87±0.37 μm), and better than carboplatin (IC50=14.75±0.38 μm), which was a confirmation of the hypothesis that the rate of hydration is crucial to the efficiency (Fig.13). Similar efficacy was observed when the inventors replaced glycosamin Ethylenediamine, creating similar glucosamine environment complexation of Pt (Fig.13A). It is, moreover, confirmed by the observation that the nanoparticles PIMA-GA-cisplatin (N->Pt) (1C 50=6.36±0.19 μm) were significantly less active than cisplatin, suggesting that the environment of platinum is critical to determine the rate of hydration. To further assess the role of environment complexing the inventors have created PIMA-GA(20), in which only 20 out of 40 monomers comprising the polymer PIMA were derivatization glucosamine, thereby introducing dicarboxylato communication and reducing monocarboxylate and coordination, forming a complex with Pt PIMA-GA. As shown in Fi is.16F, curve concentration-efficiency is shifted to the right with PIMA-GA(20)-cisplatin (ES=5.85±0.13 µm) compared with nanoparticles PIMA-GA-cisplatin (O->Pt), where all 40 monomers derivatization glucosamine. The unloaded polymer PIMA-GA no effect on cell viability. The table shows the values ES.

As shown in Fig.13A, while the polymer taken alone induces cell death at very high concentrations, the formation of the complex with cisplatin significantly shifts the curve concentration-effect left, which indicates that the nanoparticles PIMA-cisplatin induces cell death. However, even at a concentration of 50 μm PIMA-cisplatin does not cause complete cell death. In contrast, cisplatin causes complete cell death at a concentration of more than 20 μm. This decrease in the efficiency of platinate after complexation with PIMA can be explained dicarboxylato communication between platinum and monomers of maleic acid, which binds Pt, similar to the relationship in carboplatine, which similarly is less effective than cisplatin.

Staining of cells for the expression of phosphatidylserine on the cell surface showed that treatment with cisplatin can induce apoptopic cell death, with LLC cells are more sensitive than cells T (Fig.14).

Table 1:Values AS for different complexesAS (µm)PIMA30: PIMA-GA-Cisplatin [sour]5.29±0.11PIMA30: PIMA-GA-Cisplatin [alkaline]6.84±0.14Cisplatin3.87±0.37Carboplatin14.75±0.38PIMA40-200: PIMA-GA-Cisplatin [sour]4.25±0.16PIMA40-200: PIMA-GA-Cisplatin [alkaline]6.36±0.19RMA-GA20-Cisplatin [sour]5.85±0.13

Example 7 - active Release of cisplatin from the nanoparticles as a pH-dependent

Provided that the nanoparticles are localized in the lysosomal compartment, the inventors have tested the release of Pt nanoparticles at pH 5.5, mimicking the acidic pH endolysosomes compartment tumors (Lin, et al., Eur. J. Cancer, 2004 40(2):291-297). Also, the inventors chose pH 8.5 as the pH of the comparison in the alkaline range. As shown in Fig.16, when rn.5 nanoparticles PIMA-GA-cisplatin (O->Pt) cause a slow, but significant is the amount of release of cisplatin, observed during the 70-hour period. On the contrary, the release when rn.5 was significantly lower, indicating a pH-dependent release of Pt. Surprisingly, PIMA-GA-cisplatin (N->Pt) released significantly lower amounts of Pt even when rn.5, which is consistent with the fact that N->Pt coordination bond is stronger than O->Pt link. As expected, the inventors have observed that the nanoparticles PIMA-cisplatin showed a significantly lower rate of release of Pt compared to PIMA-GA-cisplatin (N->Pt) and PIMA-GA-cisplatin (O->Pt), since Pt was kept more stable dicarboxylato links instead monocarboxylate and coordination bonds.

Example 8 - Nanoparticles induce slower growth and regression of tumors with reduced nephrotoxicity

As nanoparticles PIMA-GA-cisplatin (O->Pt) showed the required speed of release for platinum, and also demonstrated the in vitro efficacy comparable with cisplatin, the inventors have established therapeutic efficacy of the nanoparticles in vivo. Mice bearing established Lewis lung carcinoma or breast cancer T, were randomly divided into five groups, respectively, and each group was treated with the following doses three times: (i) PBS (control); (ii) Cisplatin (1.25 mg/kg); (iii) Cisplatin (3 mg/kg); (iv) nanoparticles PIMA-GA-CIS is the latina (O-> Pt) (1.25 mg/kg); (v) nanoparticles PIMA-GA-Cisplatin (O->Pt) (3 mg/kg). Mouse injected PBS, formed large tumors on day 16 (day after the last injection) and, therefore, were manasarovara. Animals in the other groups was also killed at the same time, for evaluating the effect of treatment on the pathology of the tumor. As shown in Fig.5, cisplatin induced a dose-dependent inhibition of the tumor and the dose equivalent to 1.25 mg/kg of cisplatin, the introduction of the composition of the nanoparticles resulted in greater inhibition of progression of lung carcinoma in comparison with the free drug. However, if the dose equivalent to 3 mg/kg, free cisplatin caused a significant decrease in body weight, indicating a systemic toxicity. In contrast, animals treated with nanoparticles, equivalent to 3 mg/kg cisplatin, showed an increase in weight, although the inhibition of the tumor was similar in both treated groups (data not shown). In addition, the autopsy showed that the processing of free cisplatin causes a significant decrease in the weight of the kidney and spleen (Fig.5D and 5E), indicating neurotoxicity and hematologic toxicity, comparable with previous findings. Surprisingly, the nanoparticles of cisplatin have no effects on the weight of the kidneys and reduce the size of the spleen only at the highest doses (Fig.5D 5E). It was also installed with pathological analysis of staining H&E, transverse sections of the kidneys, which showed a significant tubular necrosis in animals treated with free cisplatin compared with cisplatin nanoparticles. To clarify the mechanism underlying the inhibition of the tumor, the inventors have noted cross sections of the tumor for TUNEL analysis, which found a significant induction of apoptosis after treatment of free cisplatin and nanoparticles PIMA-GA cisplatin (O->Pt) (data not shown). Surprisingly, the staining of sections of kidneys for TUNEL analysis showed significant apoptosis in animals treated with free cisplatin, as opposed to minimal nephrotoxicity in the group treated with nanoparticles (data not shown). Of course, studies of bearsdley using mass spectrometry with inductively coupled plasma (ICP) found that the concentration of Pt in the kidney after injection of the nanoparticles of cisplatin 50% concentration achieved after the introduction of the free drug (Fig.5E), which may explain the decrease in nephrotoxicity.

Treatment with cisplatin (1.25 mg/kg) showed only minimal inhibition of tumor growth compared to control; on the contrary, treatment with nanoparticles of cisplatin at the same dose showed mean the local increase antitumor efficacy (Fig.5A). This is consistent with the fact that nanoparticles significantly higher concentrations of active agent to remain within the tumor compared with the free drug. At higher dose of the antitumor efficacy of free drugs and nanoparticles is similar (Fig.5A), possibly representing theoretical limit for drugs. However, the free drug at this dose results in more than a 20% loss of body weight (Fig.5B), which is an indicator of non-specific toxicity. Of course, this causes a significant nephrotoxicity, as shown by the weight loss of the kidney (Fig.5D). In addition, despite the fact that the blood did not differ in different treated groups (Fig.5C), was attended by a significant loss of spleen weight at the highest dose of free cisplatin (Fig.5E). In contrast, the nanoparticles of cisplatin did not show such toxicity even at the highest dose, making it possible dosing at higher levels or for longer periods of time, which may have a significant effect on the antitumor results. In addition, ease of manufacturing, low cost materials and improve therapeutic efficacy and reduce toxicity may be an example of the m nanotechnologies, impact on world health. Without being bound by theory, increased therapeutic index may be due to the preferential accumulation of nanoparticles in tumors due to the well-studied EPR effect and does not affect the kidney because it exceeded the size limit for clearance, which, as was shown in the previous study, is less than 5 nm (Choi HS, Liu W, Misra P, Tanaka E, Zimmer JP, Itty Ipe B, Bawendi MG, Frangioni JV. Renal clearance of quantum dots. Nat Biotechnol. 2007; 25:1165-70).

Free cisplatin and nanoparticles PIMA-GA-cisplatin(O->P1) caused similar levels of inhibition of tumor growth in models of breast cancer T (Fig.17). Surprisingly, 1.25 mg/kg and 3 mg/kg of free cisplatin induced significant weight loss compared with the group treated with cisplatin nanoparticle. According to the observation model of lung cancer, while free cisplatin induced significant apoptosis in the kidney, the group treated with cisplatin nanoparticle showed minimal apoptosis in the kidney, but significant levels of apoptosis in the tumor.

In addition to the models of lung cancer and breast cancer, the inventors also evaluated the nanoparticles PIMA-GA-cisplatin(O->P1) in the model of ovarian cancer. Epithelial ovarian cancer is a deadly malignant neoplasm of the female reproductive cycle. The discovery of frequent soma is practical mutations in the PTEN gene and loss of heterozygosity in the 10q23 locus of PTEN in endometrioma ovarian cancer includes the key role of PTEN in the etiology of this subtype of epithelial ovarian cancer (Obata, K. et al. Frequent PTEN/MMAC1 mutations in endometrioid but not serous or mucinous epithelial ovarian tumors. Cancer Res. 58, 2095-2097 (1998) and Sato, et al., Cancer Res. 2000, 60: 7052-7056 and Sato, et al., Cancer Res. 2000, 60: 7052-7056). Similarly, the K-RAS oncogene is also mutated in endometrioid ovarian cancer, albeit at a lesser frequency (Cuatrecasas, et al.. Cancer (1998) 82: 1088-1095). In a recent study it was found that the combination of these two mutations in the surface epithelium of the ovary induces invasive and widely metastatic endometrioid adenocarcinoma of the ovary with full penetrance, making it a good model to simulate the progression of human tumors. In this transgenic model is processed by a native animals showed a rapid progression of the tumor, which was quantitatively assessed by the level of expression of luciferase. Processing of nanoparticles of cisplatin caused a dose-dependent inhibition of tumor progression with a lower dose, equivalent to 1.25 mg/kg, causing inhibition similar dose of 3 mg/kg of free cisplatin (Fig.18). The higher dose of cisplatin nanoparticles (equivalent to 3 mg/kg cisplatin) resulted in greater inhibition of tumor without significant loss of body weight observed at the same dose of free cisplatin, which is approved for clinical use in cancer of the ovary (Fig.18). In addition, TUNEL staining showed significant apoptosis in the kidney at 3 mg/kg of free cisplatin in thatduring nanoparticles of cisplatin at equivalent concentrations of Pt is not induced apoptosis nephrons.

Example 9 - the Biodistribution of the nanoparticles of cisplatin after repeated administration of doses

To study bearsdley nanoparticles of cisplatin, the inventors removed the tumor at the end of the experiments repeated the dose in which each animal received three doses of free drug or nanoparticles of cisplatin. As shown in Fig.19, was attended by the preferential accumulation of Pt in tumors of the breast and ovary in the introduction in the form of nanoparticles, unlike the case of delivery in the form of free cisplatin.

Example 10 Assessment of toxicity processing PIMA-GA-oxaliplatin

As can be seen in Fig.23C, when the dose of 15 mg/kg of free oxaliplatin all animals died due to systemic toxicity. On the contrary, in the case of nanoparticles oxaliplatin toxicity was absent even at this dose.

Example 11 - scheme of the synthesis of the conjugate lipid-cisplatin

In addition to the conjugate PMA-GA-cisplatin, the inventors have also developed similar, in which maleic acid is conjugated with the end of PEG pegylated lipid (PEG2000-cells of the dspe). The inventors formed a complex with Pt maleic acid, which caused the formation of platinized lipid derivative, in which Pt is a hydrophilic end and lipid forms a hydrophobic end. It forms micelles in water, and the efficiency naked is narrow is 45 μg/mg lipid derivative. This can be increased through the use of PEG lower molecular weight or lipid. See Fig.10.

Example 12 - Cisplatin-lipogenetic

Materials and method

All reactions were performed under inert conditions, unless otherwise noted. All commercially obtained compound was used without further purification. DCM, dry DCM, methanol, cholesteryl chloroformate, cholesterol, Ethylenediamine, succinic anhydride, silver nitrate, sodium sulfate, pyridine, cisplatin, L-a-phosphatidylcholine, Sephadex G-25 and 1,2-phenylenediamine were obtained from Sigma-Aldrich. 1,2-Distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino) polyethylene glycol)2000], and handheld mini Extruder set (includes membrane filter of 0.2 μm Whatman Nucleopore Track-Etch Membrane, the support of Whatman filter and 1.0 ml syringes Hamilton) was purchased from the company Avanti Polar Lipids Inc. Anhydrous DMF solvent was obtained from Acros Organics. Postwarranty acid was obtained from Ted Pella, Inc. Analytical thin-layer chromatography (TLC) was performed using plates of silica gel 60 F254 on aluminum substrate obtained from EMD Laboratories. Spots on TLC plates were visualized with permanganate of an alkali metal or 6% solution of ninhydrin in acetone. 1H NMR spectra (300 MHz) were obtained on spectrophotometre Varian Mercury 300. Chemical shifts are expressed in parts per million (M. D.) the use of suitable deuterated solvents for NMR spectroscopy relative to TMS at About M. d.. The MTS reagent was obtained from Promega. Analysis of the viability of the cells and the data of the kinetics of release were plotted on a graph using the software GraphPad Prism. The analysis of each sample was done in three replications.

Synthesis of (11):

1044 ál (15 EQ.) Ethylenediamine (12) was dissolved in 5.0 ml anhydrous DCM and then cooled to 0-5°C with ice. 500.0 mg (1.0 EQ.) cholesterol of chloroformate was dissolved in 5.0 ml anhydrous DCM was added to the reaction mixture dropwise over 15 minutes with vigorous stirring and the stirring was continued throughout the night until, until the temperature will be room temperature. The reaction mixture was treated with water (50 ml×3) and DCM (50 ml), followed by washing with saturated saline solution. The organic layer was dried over anhydrous sodium sulfate and evaporated using a rotary evaporator. Pure oily product is light yellow (13) was shared with the release of 99.1%. 1H-NMR (300 MHz) d(M. a.)=5.37 (s, 1H), 5.06 (S, 1H), 4.49 (bs, 1H), 3.22-3.20 (m, 2H), 2.82-2.81 (m, 2H), 2.34-2.26 (m, 2H), 2.02-1.83 (m, 6H), 1.54-0.84 (m, 37H).

Synthesis of 15:

350 mg (0.74 mmol, 1 EQ.) source material (13) was dissolved in 5.0 ml anhydrous DCM. Added 370.0 mg (3.7 mmol, 5 EQ.) succinic anhydride (14) and a catalytic amount of pyridine. Was stirred for 1 day followed by treatment with 0.1 (N) HC1 and DCM several times. The organic layer was dried the hell sodium sulfate and evaporated to yield a white amorphous solid compounds (15). Yield: 95%. 1H-NMR (300 MHz) d(M. D.) - 7.72-7.70 (m, 1H), 7.54-7.53 (m, 1H), 5.37 (s, 1H), 5.07 (s, 1H), 4.49 (bs, 1H), 4.22-4.19 (m, 2H), 3.36-3.30 (m, 4H), 2.68-2.33 (m, 4H), 2.02-1.83 (m, 6H), 1.54-0.84 (m, 37H).

Synthesis of 16:.

50 mg (0.166 mmol, 1 EQ.) cisplatin (16) was partially dissolved in 10.0 ml of N2The acting was Added 28.0 mg (0.166 mmol, 1 EQ.) silver nitrate and the resulting reaction mixture was stirred at room temperature for 1 day. The mixture looked milky-white and silver chloride was removed by centrifugation at 25000 X g for 1 h the resulting product 5 in the ratio 7:200 mg (0.35 mmol, 1.0 EQ.) was dissolved in 5.0 ml of DMF. Added 20.0 ml of product 6 (conc. 5.0 mg/ml, 1.0 EQ.) and was stirred for 1 day. The reaction mixture was dried using lyophilizate. The dried product (17) was used for the synthesis of lipogenetic without further purification.

General methods of synthesis of lipogenetic:

10.0 mg L-a-phosphatidylcholine, 5.0 mg of cholesterol (or conjugate of Pt(II)-cholesterol) and 1.0 mg 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-M-[amino(polyethylene glycol)2000] was dissolved in 10.0 ml of DCM. The product is evaporated in a thin and uniform film using a rotary evaporator. After thorough drying using a pump was hydrational 1.0 ml of H2O for 2 h at 60°C. Hydrated lipogenetic had color from light yellow to white, with a slightly viscous texture. The product was passed through a column of Sephadex G-25 and extras who were denovali at 65°C.

General method for quantitative estimation of Pt(II) lipogenetic

A measured quantity of extruded lipogenetic was heated at 100°C 1.2 mg/ml 1,2-phenylenediamine in DMF for 2 hours Number of Pt(II) was calculated using UV spectrophotometry (Shimadzu 2450).

The kinetics of release

Concentrated loaded drug of lipogenetic suspended in the buffer (or cell lysate) and tightly closed in a dialysis membrane (cutoff molecular weight of 1000, Spectrum Lab). Dialysis bags were incubated in 1.0 ml of PBS buffer at room temperature with vigorous shaking. An aliquot of 10 µl were taken from the incubation medium at pre-defined intervals of time and the released drug was quantitatively evaluated using a UV spectrophotometer (Shimadzu 2450). The results were plotted on the graph as the percentage of release.

Sample preparation for THE

The high-resolution images were obtained on a high-contrast digital THE Jeol 2011. For preparation of the sample is covered with a Lacy carbon copper mesh 300 mesh (Electron microscopy Science) was immersed in an aqueous solution of lipogenesis. After air drying were stained with 2% aqueous solution phospholipases acid. Distribution of lipogenetic size was investigated by dynamic scattering St is the (DLS), which was performed at 26°C on DLS system Malvem Zetasizer, equipped with a He-Ne laser.

Analysis of viability of cells

Sown 2×103cells in 96-well plates. After 4 h the cells were treated with different concentrations of free drug or lebanonization. Cells without treatment were kept as control. After 48 h cell viability was assessed using a standard MTS analysis according to the manufacturer's instructions.

The study of in vivo efficacy and toxicity

Mouse strain BALB/c mice were inoculated with tumor cells of the breast T subcutaneously in the amount of 1×105in 100 μl of PBS into the right side. The processing of different anticancer agents, free or incarcerated in nanoparticles, began on the day when tumor volume reached 200 mm3. As a rule, the animals received only the free drug or nanoparticles by intravenous injection through the day in total, equal to three doses. Once tumor volume reached 2000 mm3in the control group, mice were killed. The tumor, kidneys, spleen, lungs, livers were removed and processed to fill in paraffin and cooking of the slices.

All patents and publications here, incorporated herein fully by reference.

1. Biocompatible conjugated polymerasechain, including:
the main chain of the copolymer, and at least one monomer of the polymer contains two side chains selected from the group consisting of carboxylic acid, amide and ether complex, and these side chains are separated from each other 1-10 carbon atoms, oxygen atoms, or sulfur atoms, or any of their combinations;
many of the side chains covalently associated with the specified main chain, and these side chains selected from the group consisting of monosaccharides, dicarboxylic acids, polyethylene glycol (PEG), and combinations thereof; and
many platinum compounds, dissociatio associated with the specified primary circuit, where at least one of a variety of platinum compounds is associated with the specified main chain through at least one coordination bond between the carbonyl oxygen of the carbonyl or amide group in the main chain and the platinum atom of the platinum compounds, and specified the platinum compound is selected from compounds of Pt(II) compounds of Pt(IV) and any combinations thereof.

2. The nanoparticles under item 1, in which the main chain contains at least one monomer of the formula-CH(CO2H)-R-CH(C(O)R')-; -CH(CO2H)-R-CH(C(O)R')CH2C(CH3)2- or-CH(C(O)R')-R-CH(CO2H)-CH2C(CH3)2- where R is a bond or alkylene, with alkylene may include one or more double or triple the CBE is she; and R' is a substituted nitrogen atom.

3. The nanoparticles under item 1, in which the specified copolymer contains the monomer of maleic acid.

4. The nanoparticles under item 2, in which from 50% to 100% of the monomers in the main chain represented by the formula-CH(CO2H)-R-CH(C(O)R')-; -CH(CO2H)-R-CH(C(O)R')CH2C(CH3)2- or-CH(C(O)R')-R-CH(CO2H)-CH2C(CH3)2.

5. The nanoparticles under item 1, in which the specified platinum compound is a compound of Pt(II) selected from the group consisting of cisplatin, oxaliplatin, carboplatin, paraplatin, satraplatin and their combinations.

6. Biocompatible conjugated polymer nanoparticles under item 1, in which the copolymer is a poly(isobutylene-alt-maleic acid) containing from 25 to 50 Monomeric units; many of the side chains of PEG covalently associated with the main chain having a molecular weight of from 1000 to 3000 daltons, and a specified number of PEG side chains is from 50% to 100% of the number of monomer units specified the main polymer chain; and the number of lateral groups of cisplatin is from 25% to 75% of the number of monomer units specified the main polymer chain.

7. Biocompatible conjugated polymer nanoparticles under item 1, in which the copolymer is a poly(isobutylene-alt-maleic acid) containing from 25 to 50 Monomeric units; m is these side chains covalently linked with the main chain are glucosamine, and their number ranges from 50% to 100% of monomer units of the main polymer chain; and the number of these side groups of cisplatin is from 25% to 75% of the number of monomer units specified the main polymer chain.

8. Biocompatible conjugated polymer nanoparticles including:
the complex carboxylic acid-platinum compound, in which the specified carboxylic acid represented by the formula HOOC-R-COOH, where R is a bond or C1-C6alkyl, C2-C6alkenyl or2-C6quinil and where specified the platinum compound is selected from compounds of Pt(II) compounds of Pt(IV) and any combinations thereof; and
many lipid-polymer chains, while the carboxylic portion of the specified complex carboxylic acid-platinum compound is covalently associated with these lipid-polymer chains, where the platinum compound in the complex is associated with a carboxylic acid through at least one coordination bond between the carbonyl oxygen of the carbonyl of the acid and the platinum atom of the platinum compounds.

9. The nanoparticles under item 8, in which the carboxylic acid is maleic acid.

10. The nanoparticles under item 8 in which the polymer is PEG.

11. The nanoparticles under item 8, in which the platinum compound is a compound Pt(II) selected from the group consisting of cisplatin,oxaliplatin, carboplatin, paraplatin, satraplatin and their combinations.

12. The nanoparticles under item 8, in which the load of a platinum compound is from 1% to 30%.

13. Dicarbonyl-lipid compound having the structure

14. The compound in the form of vesicles, micelles or liposomes containing a variety of nanoparticles, including dicarbonyl-lipid connection on p. 13 and a platinum compound, where the platinum compound is dissociatio associated with the connection on p. 13, and where specified, the platinum compound is selected from compounds of Pt(II) compounds of Pt(IV) and any combinations thereof.

15. Connection on p. 14, characterized in that said platinum compound is a compound Pt(II) selected from the group consisting of cisplatin, oxaliplatin, carboplatin, paraplatin, satraplatin and their combinations.

16. The nanoparticles under item 4, in which at least 90% of the monomers in the main chain represented by the formula-CH(CO2H)-R-CH(C(O)R')-; -CH(CO2H)-R-CH(C(O)R')CH2C(CH3)2- or-CH(C(O)R')-R-CH(CO2H)-CH2C(CH3)2-.

17. The nanoparticles under item 2, in which R' represents

18. The nanoparticles under item 2, further comprising a bioactive agent.

19. A method of treating cancer or metastasis, including:
introduction to a subject in need, an effective amount nanocast the hospitals under item 1.

20. The nanoparticles under item 1, characterized in that at least one of a specified set of platinum compounds is associated with the side chain at least one coordination bond, where this coordination is between the oxygen of the side chain and the platinum atom of the platinum compounds.

21. Biocompatible polymer, including:
the main chain of the copolymer, and at least one monomer of the polymer contains two side chains selected from the group consisting of carboxylic acid, amide and ether complex, and these side chains are separated from each other 1-10 carbon atoms, oxygen atoms, or sulfur atoms, or any of their combinations;
many of the side chains covalently associated with the specified main chain, and these side chains selected from the group consisting of monosaccharides, dicarboxylic acids, polyethylene glycol (PEG), and combinations thereof; and
many platinum compounds, dissociatio associated with the specified primary circuit, where at least one of a specified set of platinum compounds is associated with the main chain at least one coordination bond, which is located between the oxygen of the main chain and the platinum atom of the platinum compounds, and specified the platinum compound is selected from compounds of Pt(II) compounds of Pt(IV) and any of their combination is s.

22. Biocompatible polymer under item 21, in which the specified oxygen is the carbonyl oxygen of the carbonyl or amide group.

23. Biocompatible polymer under item 21, in which the main chain contains at least one monomer of the formula-CH(CO2H)-R-CH(C(O)R')-; -CH(CO2H)-R-CH(C(O)R')CH2C(CH3)2- or-CH(C(O)R')-R-CH(CO2H)-CH2C(CH3)2- where R is a bond or alkylene, with alkylene may include one or more double or triple links; and R' is a substituted nitrogen atom.

24. Biocompatible polymer under item 23, in which the platinum compound is a compound of Pt(II) selected from the group consisting of cisplatin, oxaliplatin, carboplatin, paraplatin, satraplatin and their combinations.

25. Biocompatible polymer under item 21, wherein said copolymer contains from 2 to 100 monomer units in the main chain.

26. Biocompatible polymer under item 21, in which the main chain of the copolymer is a poly(isobutylene-alt-maleic acid).

27. Biocompatible polymer under item 21, in which from 50% to 100% of the monomers in the main chain represented by the formula-CH(CO2H)-R-CH(C(O)R')-; -CH(CO2H)-R-CH(C(O)R')CH2C(CH3)2- or-CH(C(O)R')-R-CH(CO2H)- CH2C(CH3)2-.

28. Biocompatible polymer according to p. 27, in which at least 90% of the monomers in the main the ETUI is represented by the formula-CH(CO 2H)-R-CH(C(O)R')-; -CH(CO2H)-R-CH(C(O)R')CH2C(CH3)2- or-CH(C(O)R')-R-CH(CO2H)- CH2C(CH3)2-.

29. Biocompatible polymer under item 21, in which the number of side chains is from 50% to 100% of the number of monomer units specified the main polymer chain.

30. Biocompatible polymer under item 21, in which the specified number of platinum compounds is from 10% to 100% of the number of monomer units specified the main polymer chain.

31. Biocompatible polymer according to p. 21, including:
the main chain, a poly(isobutylene-alt-maleic acid), which contains from 25 to 50 Monomeric units;
many polietilenglikolya (PEG) or glucosamine side chains covalently associated with the specified main chain, with PEG side chains have a molecular weight of from 1000 to 3000 daltons and the number of PEG side chains or glucosamine side chains is from 50% to 100% of monomer units of the specified main polymer chain, and
many of the side groups of cisplatin, dissociatio associated with the specified main chain, the number of these side groups of cisplatin is from 25% to 75% of the number of monomer units specified the main polymer chain.

32. Biocompatible polymer under item 21, which is:

33. Conjugate, including:
dicarbonyl molecule, where the platinum compound is dissociatio associated with dicarbonyl molecule R OC(O)-R,-C(O)-, where R represents a C1-C6alkylen where alkylene may contain one or more double or triple links, and/or the main chain alkylene may be interrupted by one or more of O, S, S(O), SO2, NH, C(O); and R' represents H, alkyl, alkenyl, quinil, aryl, heteroaryl, cilil, heterocyclyl, each of which may be optionally substituted, and where specified, the platinum compound is selected from compounds of Pt(II) compounds of Pt(IV) and any combinations thereof; and
lipid molecule, and dicarbonyl molecule covalently linked to a lipid molecule.

34. Conjugate under item 33, in which dicarbonyl molecule is succinic acid.

35. Conjugate under item 33, in which dicarbonyl molecule covalently linked to a lipid molecule through a linker.

36. Conjugate under item 33, in which the platinum compound is associated with dicarbonyl molecule via a coordination bond, which is between the carbonyl oxygen molecule and atom of platinum of the platinum compounds.

37. Conjugate under item 33, in which the platinum compound is a compound of Pt(II) selected from the group consisting of cisplatin, oxaliplatin, carboplatin, paraplatin SAR is replacin and their combinations.

38. Conjugate under item 33, which is:

39. Biocompatible polymer under item 21, in which these side chains selected from the group consisting of carboxylic acid, amide, complex ether, separated from each other by two carbon atoms.

40. The nanoparticles under item 1, in which the mentioned side chains selected from the group consisting of carboxylic acid, amide, complex ether, separated from each other by two carbon atoms.



 

Same patents:

FIELD: chemistry.

SUBSTANCE: claimed is a method of obtaining borated 1,2-polybutadienes, consisting in the interaction of a polymer with a borating agent, characterised by the fact that before the stage of borating aluminisation and hydroxylation of 1,2-polybutadiene are carried out, where as an aluminising agent used is a mixture of diisobutylaluminium chloride in the presence of (i-OPr)4Ti with a molar ratio of 1,2-polybutadiene:diisobutylaluminium chloride=1:0.2-4, 1,2-polybutadiene:(i-OPr)4Ti=1:0.01-0.02 at a temperature of 10-55°C for 12-26 h, hydroxylation of the aluminium derivative of 1,2-polybutadiene is carried out by the action of a hydroxylating agent, which is represented by air oxygen, by hydroxylation of the reaction mass at room temperature for 7-20 h, after which borating is carried out by means of the borating agent, consisting of boric acid and a sodium hydroxide solution, with a molar ratio hydroxylated 1,2-polybutadiene:boric acid 1:0.2-1.3, boric acid:sodium hydroxide 1:0.5-4.2 at room temperature for 0.5-4 h.

EFFECT: reduction of energy consumption and elimination of toxic reagents from the process of synthesis.

2 cl, 1 tbl, 17 ex

FIELD: chemistry.

SUBSTANCE: disclosed is a method of producing a modified polymer based on a conjugated diene, involving a step (a) of conducting a reaction between a silicon compound and a conjugated diene-based polymer having an active group terminal group so that the reaction takes place on the terminal group, wherein the silicon compound in its molecule contains a protected primary amine group which is removed by hydrolysis to obtain a primary amine group, and a bifunctional silicon atom bonded to a hydrocarbyloxy group and a reactive group, which modifies the active terminal group, and a step (b) for carrying out a condensation reaction which involves participation of the compound which contains a bifunctional silicon atom in the presence of a titanium compound which is used as a titanium-based condensation accelerator. The reactive group at the silicon atom is a hydroxycarbyloxy group which contains 1-20 carbon atoms. Also disclosed is a modified polymer obtained using the disclosed method, a rubber composition containing said polymer and a pneumatic cover using the disclosed composition.

EFFECT: method enables to obtain a polymer which has excellent operational properties.

13 cl, 9 tbl, 30 ex

FIELD: chemistry.

SUBSTANCE: invention relates to synthesis of water-soluble trimetallic salts of acrylic and methacrylic acid copolymers. The method of producing water-soluble trimetallic salts of acrylic and methacrylic acid copolymers involves polymerisation of a mixture of acrylic and methacrylic acid in the presence of potassium persulphate with subsequent addition of a mixture of aqueous solutions of chlorides or sulphates of di- or trivalent metals in amount of 6.802-35.2776 moles and neutralisation of the remaining free acid with aqueous sodium hydroxide solution.

EFFECT: obtaining water-soluble salts of copolymers of different composition.

1 cl, 20 ex

FIELD: chemistry.

SUBSTANCE: invention relates to metal-polymer complex of europium (Eu3+) and (co)poly-(methylmethacrylate)-(1-methacryloyl-2-(2-pyridyl)-4-carboxy quinolyl)hydrazine of formula: where: m:k = 80 - 95.5:20 - 3.9:0 - 0.6 mol %, MM from 17000 to 24000 Da, Lig denotes a low-molecular weight ligand selected from dibenzoyl methane, thenoyltrifluoroacetone, having Eu3+ ion content from 2.6 to 9.6 wt %.

EFFECT: obtaining complexes with high luminescence intensity.

2 cl, 10 ex, 1 tbl, 2 dwg

FIELD: chemistry.

SUBSTANCE: invention relates to lanthanide-containing compounds, consisting of a copolymer of ethyl methacrylate and 3-allylpentanedione-2,4 (100:1), bonded through a β-diketonate group with a lanthanide (+3) ion, which in turn is bonded with molecules of a ligand which is β-diketone of general formula: , where Ln is a lanthanide (+3) ion (La3+, Pr3+, Nd3+, Sm3+, Eu3+, Gd3+ Tb3+, Dy3+, Ho3+, Er3+, Tm3+, Yb3+), n is the number of ethyl methacrylate links in the copolymer chain; m is the number of lanthanide-containing links in the copolymer chain; R1, R2, R3, R4 are organic radicals (CH3 - methyl, C6H5-phenyl): - R1=R2=R3=R4=CH3 - lanthanide (+3) ion, bonded with the polymer part of the compound through a pentanedione-2,4 fragment (acetyl acetone) and a ligand which is acetyl acetone; - R1 = R3 = CH3, R2=R4=C6H5 - lanthanide (+3) ion bonded with the polymer part of the compound through benzoyl acetone fragment and a ligand which is benzyol acetone; - R1=R2=R3=R4=C6H5 - lanthanide (+3) ion bonded with the polymer part of the compound through a dibenzoyl methane fragment and a ligand which is dibenzoyl methane; - R1=R3=R4=CH3, R2=C6H5 - lanthanide (+3) ion bonded with the polymer part of the compound through a benzoyl acetone fragment and a ligand which is acetyl acetone; - R1=R2=C6H5, R3=R4=CH3 - lanthanide (+3) ion bonded with the polymer part of the compound through a dibenzoyl methane fragment and a ligand which is acetyl acetone; - R1=R2=R3=C6H5, R4=CH3 - lanthanide (+3) bonded with the polymer part of the compound through a dibenzoyl methane fragment and a ligand which is benzoyl acetone. Said compounds have radiation-protective properties.

EFFECT: obtaining material which is optically transparent in the visible and ultraviolet range and resistant to ionising radiation.

1 ex, 1 tbl, 9 dwg

FIELD: medicine.

SUBSTANCE: described is incomplete cesium salt polyacrylic acid characterised by the fact that it corresponds to formula

Also described is method of obtaining said incomplete cesium salt of polyacrylic acid, characterised by the fact that to water solution of polyacrylic acid added is water solution of cesium chloride, reaction mixture is mixed for 30-40 minutes, obtained water solution is passed through cationite and dried in vacuum at temperature not higher than 50 degrees. Described is medication, possessing hemostatic action if applied externally, containing i.1 substance, characterised by the fact that it made in form of solution with ratio from 1- 10 g of basic substance per 100 ml of water.

EFFECT: obtaining novel chemical compound, which can be applied in medicinal practice as hemostatic medication of local action.

3 cl, 5 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: claimed invention relates to creation of novel chemical compounds, which can be used in medicine as hemostatic medications of local action. Described in incomplete lithium salt of polyacrylic acid (liacryl), which is characterised by correespondance to formula: (-CH2-CHCOOH-)n(-CH2CHCOOLi-)m, where n=13496-28570; m=1672-4499, content of lithium constituting 6-8 wt % (0.0011-0.0012 g-at), and content of carboxyl groups constitutes 0.0118-0.0121 g-equiv per 1 g of polymer, molecular weight is 1000000-3000000. Also described is incomplete double lithium-zinc salt of polyacrylic acid (licicryl), corresponding to general formula: (-CH2CHCOOH-)n(-CH2CHCOOLi-)m(-CH2CHCOOZn-)k, where n=13777-18255; m=4227-4679, k=1666-2180, total content of metals constitutes 6-8 wt % (0.0010-0.0011 g-at), content of carboxyl groups constitutes 0.0117-0.0120 g-equiv, molecular weight is 1000000-3000000. Described are methods of obtaining said above incomplete salts of polyacrylic acid. Also described is medication, possessing efficient hemostatic action in external application, which contains said above incomplete lithium salt of polyacrylic acid, characterised by the fact that it is made in form of solutions with ratio 1-10 g of principal substance per 100 ml of water (1-10%). Described is medication, which possesses efficient hemostatic and expressed antiseptic action in external application, which contain said above incomplete lithium-zinc salt, characterised by the fact that it is made in form of solution with ratio 2-5 g of principal substance per 100 ml of water (2-5%).

EFFECT: obtaining novel chemical compounds, which can be used in medicine as hemostatic medication of local action, possessing high hemostatic effect.

6 cl, 1 tbl, 7 ex

FIELD: chemistry.

SUBSTANCE: polymer, whose chain ends are modified, contains a product of reacting a living anionic elastomeric polymer and a silane sulphide modifier of formula: (RO)x(R)ySi-R'-S-SiR3. The invention also relates to a vulcanised composition based on the elastomeric polymer and a method of preparing said composition. The method involves combining filler, vulcanising agent, elastomeric polymer modified on chain ends and vulcanisation of the elastomeric polymer composition. The composition is used to make articles such as pneumatic tyres, tyre protectors, belts and suchlike.

EFFECT: preserving good technological properties and good balance of physical and mechanical properties, including wear resistance, breaking strength, apparent tensile stress and breaking elongation.

23 cl, 9 tbl, 19 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a multifunctional polymer which contains a graft polymer formed from: a) polyolefin, b) nitrogen-containing ethylene-unsaturated aromatic or aliphatic monomer which contains from 2 to approximately 50 carbon atoms; and c) an organometallic compound capable of reacting with the said polyolefin. The invention also discloses multifunctional polymers (versions), a synthesis method thereof, and lubricating oil (versions).

EFFECT: invention enables to obtain a multifunctional polymer which acts as a dispersing agent, viscosity index improver and an antiwear additive.

62 cl, 7 ex, 3 tbl

FIELD: pharmaceuticals.

SUBSTANCE: polyvinyl alcohol and magnesium or calcium chlorides polyhydrated complexes are obtained by dissolution of polyvinyl alcohol and magnesium or calcium chlorides in water on boiling water bath. Said components are taken in the next ratio (mass%): polyvinyl alcohol 11.5-11.6; magnesium or calcium chloride 23.2-24.0; and balance: distilled water.

EFFECT: new complex compounds having antiinflammation, resolution and analgesic action.

11 ex

FIELD: chemistry.

SUBSTANCE: invention relates to method of obtaining cement compositions, holding sagging or holding sagging with high early strength. Method includes mixing hydraulic cement, filling agent, water and sagging-holding additive, which represent dynamic polycarboxilate copolymer. Copolymer contains residues of, at least, the following monomers: A) unsaturated dicarboxylic acid, B) at least, one ethylene-unsaturated alkenyl ether, which has C2-4 oxyalkylene chain with from 1 to 25 links, C) at least, one ethylene-unsaturated alkenyl ether, which has C2-4 oxyalkylene chain with from 26 to 300 links, D) ethylene-unsaturated monomer, which contains part, hydrolysed in cement composition, in which residue of ethylene-unsaturated monomer, when hydrolysed, contains active binding site. Ratio of component A acid monomer to alkenyl ethers of component B and component C(A):(B+C) constitutes from1:2 to 2:1. Ratio of acid monomer of component A to ethylene-unsaturated monomer of component D, which contains hydrolysed part, constitutes from 16:1 to 1:16.

EFFECT: invention makes it possible to preserve placeability of composition for a long period of time and increase compression strength.

30 cl, 5 dwg, 8 tbl, 29 ex

FIELD: medicine, polymers, pharmacy.

SUBSTANCE: invention relates to a copolymer or its pharmacologically acceptable salt that comprises the following components as elemental links forming their: (a) one or some structural elemental links describes by the formula (I) given in the invention description, and (b) one or some structural links describes by the formula (II) given in the invention description. Disposition of these structural elements represented by the formulae (I) and (II) is chosen from the following sequences: (i) sequence with alternation "head-to-head"; (ii) sequence with alternation "head-to-tail"; (iii) mixed sequence with alternation "head-to-head" and "head-to-tail"; (iv) random sequence and taking into account that the ratio between structural links of the formula (I) and structural links of the formula (II) in indicated copolymer is in the range from 10:1 to 1:10. Also, the invention relates to a copolymer or its pharmacologically acceptable salt synthesized by addition of one or some links of carboxylic acid anhydride described by the formula (III) given in the invention description that comprises as elemental links: (a) one or some structural elemental links described by the formula (I), and (b) structural link comprising carboxylic acid anhydride link described by the formula (III) for one or some reactions chosen from the group consisting of: (i) hydrolysis; (ii) ammonolysis; (iii) aminolysis, and (iv) alcoholysis. Also, invention relates to a pharmaceutical composition used for prophylaxis or treatment of osseous metabolism disorder and comprising an acceptable excipient or carrier, at least one of above indicated copolymers or their pharmaceutically acceptable salts and at least one protein representing osteoclastogenesis inhibition factor (OCIF) or its analogue, or variant. Also, invention relates to a modifying agent comprising above said copolymers, to a complex between of one of above said copolymers and protein or its analogue, or variant, to a pharmaceutical composition comprising this complex. Also, invention relates to a method for time prolongation when OCIF is retained in blood stream after intake by a patient a complex between protein and at least one of above said copolymers. Also, invention relates to a method for treatment or prophylaxis of disorders of osseous metabolism involving intake by a patient the effective amount of complex comprising complex including OCIF or its analogue or variant and bound with at least one of the claimed copolymers. Also, invention relates to use of the complex comprising OCIF bound with at least one of the claimed copolymers designated for preparing a drug designated for prophylaxis or treatment of disorder of osseous metabolism and showing sensitivity to the protein effect. Modifying the protein, namely OCIF, by the claimed copolymers results to formation of complex possessing uniform properties being especially characterizing by reduced formation of disordered structure cross-linked with protein, improved retention of the protein activity and the excellent retaining protein in blood after intake of the indicated complex.

EFFECT: improved and valuable medicinal and pharmaceutical properties of agents.

110 cl, 13 tbl, 3 dwg, 40 ex

FIELD: chemical technology.

SUBSTANCE: invention relates to a method for preparing a binding agent for a dry film photoresist of aqueous-alkaline development used in preparing a picture in making printed boards in radio- and electronic industry and as a component of paint and varnish covers and glues. Invention describes a single-step method for preparing a binding agent for a dry film photoresist of aqueous-alkaline development. At the first step method involves the copolymerization reaction of styrene with maleic anhydride in acetone medium in the presence of azo-bis-isobutyric acid dinitrile and hydroquinone at temperature 54-72°C for 2.5-3.0 h. At the second step method involves addition of 28.0-32.0 mas. p. p. of n-butanol to the reaction mixture and distilling off of 15.0-20.0 mas. p. p. of acetone followed by carrying out esterification of the copolymerization product at temperature 64-74° for 90-120 h. Prepared dry film photoresists of aqueous-alkaline development possess high stability (9-10 h) and enhanced galvanic-chemical resistance.

EFFECT: improved preparing method, improved and valuable properties of binding agent.

1 tbl, 6 ex

FIELD: organic chemistry, polymers, fertilizers.

SUBSTANCE: invention relates to new anionic, biodegradable, water-soluble polymers and their derivatives that can be used in agriculture. Invention describes a water-soluble biodegradable polymer comprising repeating dicarboxylic polymeric links consisting of at least two different fragments taken among fragment group A, B and C (see appendix). Above indicated polymeric links are formed by fragments A, B and C. Also, invention relates to a method for its preparing. Invention describes a fertilizer wherein its particles are in the tight contact with the indicated polymer. Also, invention discloses a method for improving the plant growth by using the composition comprising the indicated polymer, a method for reducing nitrogen evolving from the fertilizer by applying the indicated polymer, a method for enhancing availability of phosphorus involving a stage for applying the indicated polymer on fertilizer. Also, invention describes the composition for enhancing the plant growth, the priming product wherein the primer in the tight contact with polymer, and a method for enhancing abrasive stability of the solid fertilizer. Polymers prepared by such method show the significant effect on enhancing availability of phosphorus from ammonium-phosphorus fertilizers, promote to the plant growth enhancing and enhance the abrasive stability of fertilizer particles.

EFFECT: valuable properties of polymers.

16 cl, 7 tbl, 26 ex

The invention relates to the modification of (co)polymers of ethylene by grafting them maleic anhydride (S) in the presence of radical initiators, peroxide type

FIELD: chemistry.

SUBSTANCE: maleinised tall oil-based adhesive additive contains 10-30 wt % bound maleic anhydride. The tall oil used for maleinisation contains resin and fatty acids in weight ratio of 1.0:0.6-5.0, respectively. The adhesive additive is characterised by dynamic viscosity of 0.3-1.0 Pa·s. The additive can be easily batched and has good distribution in the polymer-bitumen composition. Use of the additive does not require premixing thereof with a thermoplastic elastomer, enabling preparation of the polymer-bitumen composition in a single step.

EFFECT: polymer-bitumen composition containing an adhesive additive, characterised by high adhesion to both alkaline and acidic mineral filler.

2 tbl, 12 ex

FIELD: medicine.

SUBSTANCE: invention refers to biotechnology, particularly to a microorganism test in various objects and media. The method provides conjugating electrically Fe0, MgFe2O4 or Fe3O4 labelled bacteria in an aqueous medium at specified parameters. Unbounded nanoparticles are separated with using a magnetic field, and a working electrode made of gold, platinum or graphite-bearing materials and having a surface pre-modified by antibodies specific to a bacterial strain to be tested is immersed into an analysed solution. The electrode is kept at the specified parameters to form an immune complex on its surface and washed in a buffer solution containing normal horse serum and Tween-20. The electrode is removed from the solution and placed into a cell containing LiClO4, dissolved in acetonitrile, dimethyl formamide or dimethyl sulphoxide; the bacterial count is determined by a value of analytical oxidation of the nanoparticles localised in the immune complex on the surface of the working electrode.

EFFECT: invention enables providing higher analysis sensitivity, better output and simplified analysis procedure.

7 dwg, 6 ex

FIELD: metallurgy.

SUBSTANCE: method of obtaining of nano-dispersed nickel-plated powders in a flow of low-temperature nitric plasma includes placing into the batcher of piston type of powdered initial reagent and its feeding by pneumatic current into the evaporator chamber, treatment in the evaporator chamber by low-temperature nitric plasma, refrigeration of the evaporation product in the nitrogen flow in water-cooled hardening chamber located in the bottom part of the evaporator, and its trapping with the filter. The initial reagent is a mix of carbide or vanadium nitride and metal nickel taken in the ratio, by wt %: carbide or vanadium nitride - 50÷75, metal nickel - 25÷50. Meanwhile the plasma temperature in the evaporator chamber is equal to 4000-6000°C, plasma flow rate is 50-55 m/s, and initial reagent is supplied with the flow rate 150-200 g/h.

EFFECT: obtaining of heterogeneous nano-dispersed nickel-plated powders of carbide or vanadium nitride, with the size of particles less than 100 nm.

6 dwg, 2 ex

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