Polymeric conjugate, the method of its production and pharmaceutical composition

 

(57) Abstract:

Usage: in medicine. The inventive polymer conjugate obtained by reacting compounds of formula II

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where one of the A2and a3means the chemical bond and the other means A, R is phenyl or t - butoxypropyl , R3means hydrogen or acetyl group, And means of simple chemical bond, amino acid residue or a peptide spacer, selected from Ala, Gly, Phe-Gly, Phe-Phe, Leu-Gly, Val-Ala, Phe-Ala, Leu-Phe, Leu-Ala, Phe-Leu-Gly, Phe-Phe-Leu, Leu-Leu-Gly, Phe-Tyr-Ala, Phe-Gly-Phe, Phe-Phe-Gly, Phe-Leu-Gly-Phe, Gly-Phe-Leu-Gly, Phe, Gly--Ala, Phe-Gly--Ala, Phe-Phe--Ala, Leu-Gly--Ala, Val-Ala--Ala, Phe-Ala--Ala, Leu-Phe--Ala, Leu-Gly--Ala, Leu-Leu-Gly--Ala, Phe-Tyr-Ala--Ala, Phe-Gly-Phe--Ala, Phe-Phe-Gly--Ala, Phe-Leu-Gly-Phe--Ala or Gly-Phe-Leu-Gly-Phe--Ala, enter into interaction with the activated polymer consisting of 90-90,9 mol.% links formula

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and N10-0,1 mol.% links formula

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the process is conducted in an anhydrous polar solvent at a temperature of from 15 to 40oC for from 1 to 24 h in the presence of organic or inorganic bases and then treated with conjugate 2-hydroxypropylamino. Pharmaceutical composition having antitumor activity, includes as an active means a polymer conjugate in efficiency is compared with paclitaxel. 4 C. and 3 h.p. f-crystals, 2 tab.

The invention is directed to associated with the polymer paclitaxel and related polymer derivatives paclitaxel with antitumor activity, in the way of their receiving and containing pharmaceuticals.

Paclitaxel (some publications also known as Taxol) belongs to taxonomy family of diterpenes, and was selected with the subsequent characterization of the extract of the bark of Taxus brevifolia L. also Known analogues of paclitaxel obtained on the basis of 10-deacetylbaccatin III, extracted from the needles of Taxus baccata L. (see Wain and others, IACS, 93, 2325 (1971); Lovelle and others , Proc. Am. Assoc. Cancer. Res., 31 (1990)). These connections, as shown, have a pronounced antitumor activity, but at the same time, poorly soluble in water, and their introduction by injection or intravenous infusion when used as a carrier of cremophor EL (trade name) followed by some toxic side effects.

The present invention provides polymer conjugant formula I consisting essentially of:

90 to 99.9 mol%. links formula

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0.1 to 5 mol%. units of the formula:

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in which one of R1R2represents estato
where R is phenyl or tert-butoxypropyl, R3is hydrogen or acetyl, A and A1that may be the same or different, represent simple chemical bond, amino acid residue or a peptide spacer, selected from Ala, Gly, Phe-Gly, Phe-Phe, Leu-Gly, Val-Ala, Phe-Ala, Leu-Phe, Leu-Ala, Phe-Leu-Gly, Phe-Phe-Leu, Leu-Leu-Gly, Phe-Tyr-Ala, Phe-Gly-Phe, Phe-Leu-Gly-Phe, Gly-Phe-Leu-Gly-Phe, Gly--Ala, Phe-Gly--Ala, Phe-Phe--Ala, Leu-Gly--Ala, Val-Ala--Ala, Phe-Ala--Ala, Leu-Phe--Ala, Leu-Gly--Ala, Phe-Leu-Gly--Ala, Phe-Phe-Leu--Ala, Leu-Leu-Gly--Ala, Phe-Tyr-Ala--Ala, Phe-Gly-Phe--Ala, Phe-Phe-Gly--Ala, Phe-Leu-Gly-Phe--Ala or Gly-Phe-Leu-Gly-Phe--Ala.

If more specifically, the present invention provides polymeric conjugates of paclitaxel and derivatives paclitaxel with improved water solubility and reduced toxicity.

It is recommended that they say. % of units containing paclitaxel and derivatives paclitaxel, 0.5-2, more preferably, if the content of pectinases in the polymer is 2-10% (wt./wt.), and the most preferred are those compounds in which the content of paclitaxel is 4-7% (wt. /wt.). The wavy line in formula means that oxygen is attached in the 7-position of the formula paclitaxel, can be in both configurations, i.e., to have -(natural) or configuration.

Content in % wt./wt. determined after enzymatic hydrolysis by HPLC according to the method similar to the method described in Cancer Treatment Rep. 71, 53-59 (1987).

Enzymatic analysis.

To 1 ml of mouse or human plasma is added to various concentrations associated with the polymer paclitaxel and at certain times (24, 48, 72, 96 h) select 100 μl and stored at -70oC until further processing.

The method of extraction.

Samples extracted by adding 75 µl of 0.5 M tetrabutylammonium (TBAF), 1250 µl CH3CN and 150 ál of 5 M NaCl and intensive shaking for 20' at 4oC.

At the end of this period the samples are centrifuged for 10' at 15000g, the supernatant collected and evaporated in high vacuum centrifuge. Samples reactivated by adding 500 μl of a mixture of MeOH-water (75:25./about.) and injection in HPLC column to determine the total content paclit the thread here - 1.5 ml/min

Detector - UV, 231 nm.

Injectiona - 20 ál.

The mobile phase - 57,5% H2O pH 2+42,5% CH3CN.

The present invention also provides a method of obtaining a polymer conjugate consisting in the reaction of compounds of formula II

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in which one of the A2and A3represents a chemical bond, and the other is A, where A, R and R3take the above values, the activated polymer comprising 90-99,9 mol.% link formula

< / BR>
and 10-0,1 mol.% link formula

< / BR>
where A1accept above values,

with the subsequent processing of the resulting polymer conjugate 2-hydroxypropylamino.

The reaction of compounds of formula II with an activated polymer is recommended in anhydrous polar organic solvent such as dimethylsulfoxide or dimethylformamide, possibly in the presence of organic or inorganic bases, such as alkali carbonate, dimethylaminopyridine or triethylamine. The reaction typically takes 1-24 hours.

The reaction is usually carried out at a temperature of 15-40oC, preferably at room temperature. Examples of the alkaline carbonate is s compounds of formula II are known compounds, for example: paclitaxel or analogues of paclitaxel, or can be obtained from known compounds.

For example, 7-EPI-derivatives can be obtained by boiling in toluene of paclitaxel or its analogues in the presence of a base (Na2CO3or databaseconnect).

Other compounds of formula II are new compounds, in particular, those compounds in which A3represents the residue is Ala, and those compounds in which either A2or A3represents a di-, tri-, or tetrapeptides the spacer according to the above definition of A, and such compounds are covered by the scope of the invention.

The compounds of formula II, in which A2different from a simple chemical bond, can be obtained by the reaction of paclitaxel or analog paclitaxel-protected amino acid or peptide in the presence of a condensing means, and in the presence or absence of a catalyst, preferably at room temperature, with subsequent removal by known methods of protective group.

Condensation can also be carried out by the use of active esters, for example, p-nitrophenylthio ether peptide or amino acids. Acceptable condentiality include: 4-dimethylaminopyridine, (DMAP), pyridine or triethylamine.

To protect the amino group can be used various known protective group, and as a source of compounds can be used, the sales are protected peptides or amino acids. Can be used amino acids or peptides, protected tert-VOS, Triticum, FMOC or carbobenzoxy (CBZ). Recommended amino acids or peptides, protected tert-VOS, Triticum and FMOC.

The compounds of formula II, in which A3different from a simple chemical bond, can be obtained by protecting or blocking 2-hydroxy-group, then tarifitsirovannyim hydroxyl at the 7-position with the subsequent removal of the protective or blocking group 2-position.

The compounds of formula II, in which A3represents the residue is Gly or Ala, more preferably be obtained by reaction of paclitaxel with 2-3 equivalents of N-protected amino acids with the formation of 2,7-disubstituted paclitaxel, the removal of amino acids in 2-position and then releasing the amino acids in the 7-position.

The reaction paclitaxel-protected amino acid is carried out in the presence of a condensing means and catalyst, similar to the above.

Cleavage receiving 2,7-di(minamikata)paclitaxel with phosphate buffer (pH 7-7.4) or a small excess of NaHCO3.

The release of amino acids is conducted according to known methods of release of amino acids, for example, treatment with a weak acid, such as acetic acid or recovery.

For example, paclitaxel leaves to react with 2-3 mol. equivalents of N-protected amino acid (tert-VOS SV or FMOC protected) in methylene chloride in the presence of DCC and catalytic amounts of 4-dimethylaminopyridine. In this way in 2 and 7-provisions impose a protected amino acid. 2,7-Bismanonline derived paclitaxel leaves 2-5 hours in the presence of NaHCO3in a mixture of H2O-MEOH, resulting in selective release 2-position with the formation of 7-substituted derivative of paclitaxel. The protective group is removed by means acceptable deblokiruyuschee agents (for example: acid, mild base or hydrogenolysis).

The activated polymer is a synthetic water-soluble polymer obtained by monopalmitate N'(2-hydroxypropyl)methacrylamide with p-nitrophenylamino esters of N-methacryloylamido according to the U.S. patent 4062831 and 4097470.

Polymer conjugates of formula I and new derivatives of paclitaxel formula II Axel in plasma or after ingestion cells by cleavage Oligopeptide spacers.

Biological activity.

Analysis on the connection and disconnection of microtubules.

Tubulin brain of a calf get in two cycles of the connection-disconnection (Ihelanski M. L. , Caskin F., Cantor C. R., RrOC. hatl. Acad. S ci. USA 70, 7665-768, 1973) and stored in liquid nitrogen in the MAV (0.1 M MES, 2.5 mm EGTA, 0.5 mm MgSO4, 0.1 mm EDTA, 0.1 mm DTT, pH 6,4).

All experiments are performed on the protein stored less than 4 weeks.

Before each experience tubulin incubated for 30 min at 4oC.

Ensembles are registered according to the method Caskin and others (Caskin F., Cantor C. R., Ihelanski M. L. Y. Molec. Biol. 89, 737-758, 1874). Cuvette (1 cm thick) with tubulin (1 mg/ml) and 1 mm GTP heated to 37oC and 340 on two dual beam spectrophotometer Perkin-Elmer 557, equipped with a recorder and a thermostatically controlled chamber for sample, continuously conduct turbidity.

After 30 minutes add 4 mm CaCl2and within 10 min the decrease in turbidity determine depolymerization. At regular intervals of 15 min add certain doses of the tested compounds and register the change in turbidity. Data expressed as percentage of repeat polymerization caused by the test compound. Obtaining the results shown in table 1.

Test the copolymer-paclitaxel obtained in example 6 in vivo against B16-P10 mouse myeloma in comparison with paclitaxel.

Mouse.

Female mice of CV obtained from the farm Charles river, Italy.

In early experiments the animal age 8-10 weeks.

Drugs.

Because of the limited solubility of paclitaxel in the water it is dissolved in nositelem 5% Tim glucose solution to the target concentration. The solution is somewhat unclear and after a short period of time observed sediment.

The compound of example 6 is easily dissolved in 5% wage glucose solution, and the resulting solution remains clear for a long time (over 2 hours). Final concentration relate to the content of paclitaxel in combination (a total of 4%).

The tumor.

Use the mouse to melanoma B16-P10. A suspension of 105tumor cells in 0.2 ml injection subcutaneously in the flank of the mouse.

Compasses measure the size of the tumor, the tumor weight calculated by the formula

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The introduction of the drug.

Paclitaxel injected intraperitoneally, due to its poor solubility and toxicity of the media.

The compound of example 6 is injected. Both compounds administered at 1, 5 and 9 days after implantation of the tumor.

Are given in table. 2 data show a higher activity of the compounds of the present invention in comparison with paclitaxel (see the end of the description).

Dose polymer conjugate refers to the content of paclitaxel.

Definition Tox. (Tox. the number of mice killed at the beginning of the poisoning) hold, if the mouse dies before control is iudenich data shows, what polymer conjugates of the present invention have excellent anti-tumor activity. Thus, these compounds are useful anticancer remedies due to their low toxicity and high solubility in water compared with paclitaxel and its analogues. Examples of tumors which are treated include: sarcoma, carcinoma, lymphoma, neuroblastoma, melanoma, tumor Wilsa, leukemia and adenocarcinoma.

Improved solubility and reduced toxicity of the polymer conjugates of the present invention means their suitability for intravenous injection or infusion. The dosage depends on age, weight and condition of the patient. The dosage may range from 1 mg/kg body weight to 1 g/kg body weight, preferably 4 to 800 mg/kg of body weight. Conventional formulations contain associated with the polymer paclitaxel/derived paclitaxel in an amount equivalent to 0,5; 1,5; 10; 20; 25 or 50 mg of active paclitaxel/derived paclitaxel.

Polymer conjugates can be prepared in the form of pharmaceutical preparations pharmaceutical acceptable carriers or diluents. Can be used with any suitable carrier or razavipour, sterile water or preferably they may be in the form of sterile water or isotonic saline solutions.

The following examples illustrate the present invention.

Example 1. Copolymer 1 methacryloylamido-2-hydroxypropane, 1-(methacryloyl-glycyl-i.e. phenylalanyl-leucyl-glycyl)amino-2-hydroxypropane and 2'-(methacryloyl-glycyl-i.e. phenylalanyl-leucyl-glycyl)paclitaxel.

To a solution of 1.4 g of copolymer 1 methacryloylamido-2-hydroxypropane and N-(methylennorbornan-Phe-Leu-Gly-4-nitrophenoxy) methacrylamide obtained by method 1. Karasek and others (Chem Makzomol. 177, 2833 (1976)), 15 ml of anhydrous dimethylformamide was added 100 mg of paclitaxel and 15 mg dimethylaminopyridine.

The yellow solution is stirred for 8 hours in anhydrous conditions at room temperature. Then the reaction flask are added dropwise 0.2 ml 2-hydroxypropylamino and the contents of the flask stirred for 30 minutes.

The reaction mixture was neutralized with 0.3 ml of glacial acetic acid, concentrated in vacuo to a small volume and then transferred into 200 ml of acetone

After stirring 30 minutes the precipitate is filtered and washed with acetone to obtain 1.25 g of the title compound.

The content of paclitaxel elaut neproreagirovavshimi paclitaxel.

Example 2. 2'-(N-Trityl-i.e. phenylalanyl-leucyl-glycyl)paclitaxel.

To a solution of 170 mg of paclitaxel in 16 ml of acetonitrile add add 24 mg dimethylaminopyridine and 150 N-trityl-i.e. phenylalanyl-leucyl-glycine-4-nitrophenylamino ether.

The yellow solution is stirred for 20 hours at room temperature, then evaporated to dryness in a vacuum. The remainder homeroot on silica gel using as eluent a mixture of ethyl acetate-hexane (35 : 25) and obtain 380 mg of the title compound.

1H-NMR (400 MHz, CDCl3) : of 0.82 (d, J = 6.4 Hz, 3H, - Leu) to 0.85 (d, J = 6,7 Hz, 3H, - Leu) and 1.15 (s, 3H, 16), of 1.26 (s, 3H, 17), of 1.2 - 1.6 (m, 3H, ++- Leu), was 1.69 (s, 3H, 19), of 1.85 (s, 1H, OH-1), 1,89 (m, 1H, 6 ), a 1.96 (d, J = 1.2 Hz, 3H, 18), and 2.14 (LW. d, J = 5,9 Hz, J = 13.5 Hz, 1H, - Phe), 2,24 (s, 3H, CH3CO/10), 2,2 - 2,7 (m, 5H, CH3-14 + OH-7 + 6+ - Phe + NH-Phe), 2,47 (s, 3H, CH3CO-4), and 3.5 (m, 1H, - Phe), 3,74 (LW. d, J = 4,7 Hz, J = 18.2 Hz, 1H, - Gly), and 3.8 (m, 1H, - Leu), 3,83 (d, J = 7 Hz, 1H, 3), 4,17 (LW. d, J = 7 Hz, J = 18.2 Hz, 1H, - Gly), 4,22, 4,33 (two d, J = 8.5 Hz, 2H, CH2-20), to 4.46 (m, 1H, 7), equal to 4.97 (LW. d, J = 2.2 Hz, J = 9.9 Hz, 1H, 5), 5,44 (d, J = 2.3 Hz, 1H, 2'), 5,71 (d, J = 7 Hz, 1H, 2), 5,97 (LW. d, J = 4,7 Hz, J = 7 Hz, 1H, NH-Gly), 6,07 (LW. d, J = 2.3 Hz, J = 9.4 Hz, 1H, 3'), 6,2 - 6,3 (m, 2H,, 13 + 10), 6,8 - 9,2 ( m, 30H, 6-Ph), to 6.95 (d, J = 6,7 Hz, 1H, NH-Leu), 8 (d, J = 9.4 Hz, 1H, NH-4').

Example 3. 2'-(i.e. phenylalanyl-leucyl-glycyl)paclitaxel.

In a mixture of ice uksusnoi temperature.

The solvent is evaporated to dryness in vacuo, the residue is stirred for 30 minutes in a mixture of diethyl ether - hexane (1 : 1) and after filtration obtain 160 mg of the title compound.

The Belarusian library Association-MS: m/z 1171/M + H/+, 1112 M-CH3COOH + 2H, 1051, 1024, 911, 603, 596, 509.

1H-NMR (400 MHz, CDCl3) : 0,88 (d, J = 6.4 Hz, 3H, - Leu), to 0.92 (d, J = 6.4 Hz, 3H, - Leu), of 1.13 (s, 3H, 16), of 1.16 (s, 3H, 17), 1,4 - 2 (m, 4H, +++ Leu + 6 ), was 1.69 (s, 3H, 19), at 1.91 (d, J = 1.2 Hz, 3H, 18), 2,16 (LW. d, J = 6 Hz, J = a 13.8 Hz, 1H, 14), of 2.23 (s, 3H, COCH3-10), 2,4 - 2,6 (m, 3H, 6 + 14 - Phe), of 2.53 (s, 3H, COCH3-4), 2,9 (LW. d, J = 4,1 Hz, J = 13.5 Hz, 1H, - Phe), 3,49 (LW. d, J = 4,1 Hz, J = 9.1 Hz, 1H, - Phe), 3,82 (d, J = 7,3 Hz, 1H, 3), 3,9 - 4,1 (m, 2H, +- Gly), 4,22, 4,33 (two d, J = 8.7 Hz, 2H, CH2-20), 4,27 (m, 1H, - Leu), of 4.44 (LW. d, J = 6,4 Hz, J = 10,8 Hz, 7), 4,98 (LW. d, J = 2.4 Hz, J = 9.7 Hz, 5), 5,61 (d, J = 3.2 Hz, 1H, 2') and 5.7 (d, J = 7,3 Hz, 1H, 2), 6,12 (LW.d, J = 3.2 Hz, J = 9.4 Hz, 1H, 3'), 6,21 (m, 2H, 13), 6,28 (c, 1H, 10), 6,8 - 8,2 (m, 21H, 4-Ph + NH Leu), 7,87 (d, J = 9.4 Hz, 1H, NH-4').

Example 4.

Copolymer 1 methacryloylamido-2-hydroxypropane, 1-(methacryloyl-glycyl)amino-2-hydroxypropane and 2'-(methacryloyl-glycyl-i.e. phenylalanyl-leucyl-glycyl)paclitaxel.

To a solution of 1 g of copolymer 1 methacryloylamido-2 - hydroxypropane and N-(metrocable-4-nitrophenoxy)methacrylamide obtained by the method of P. Rejmanova and others (Makromol. Chem. 178, 2159 - 2168), in 10 ml of anhydrous dimethylformamide dobavlyaty conditions at room temperature. Then to the reaction mixture are added 2-hydroxypropylamino (0.15 ml) and everything is stirred for 30 minutes. The reaction mixture was neutralized with 0.2 ml of glacial acetic acid, concentrated in vacuo to a small volume and transferred into 200 ml of acetone.

The mixture is stirred for 1 hour, the precipitate is filtered off and, after washing with acetone obtain 960 mg of the title compound.

The content of paclitaxel is 6% (determined by enzymatic hydrolysis and VEGS analysis).

Example 5.

Copolymer 1 methacryloylamido-2-hydroxypropane, 1-(methylacrylic)amino-2-hydroxypropane and 2'-(methacryloyl-glycyl)paclitaxel.

To a solution of 1.6 g of copolymer 1 methacryloylamido-2-hydroxypropane and N-(methylennorbornan-4-nitrophenoxy)methacrylamide in 16 ml of anhydrous dimethylformamide was added 100 mg of paclitaxel and 20 mg dimethylaminopyridine. The yellow solution is stirred for 20 hours at room temperature, then add 2-hydroxypropylamino (0.2 ml) and the mixture is stirred for 30 minutes. The reaction mixture was neutralized with 0.3 ml of glacial acetic acid, concentrated in vacuo to a small volume and then transferred into 200 ml of acetone. The mixture is stirred for 1 hour, the precipitate is filtered off and, after washing with acetone Poer 6.

Copolymer 1 methacryloylamido-2-hydroxypropane, 1-(methacryloyloxy-i.e. phenylalanyl-leucyl-glycyl)amino-2-hydroxypropane and 2'-(methacryloyl-glycyl-i.e. phenylalanyl-leucyl-glycyl--alanyl)paclitaxel.

To a solution of 620 mg of copolymer 1 methacryloylamido-2-hydroxypropane and N-(methylennorbornan-Phe-Leu-Gly-4 - nitrophenoxy)methacrylamide in 6 ml of anhydrous dimethylformamide added 62 mg of 2'-(-alanyl)paclitaxel prepared by the method of N. F. Magr. and others, (J. Nat. Products 51, 298 - 306, 1988), and 10 ml of dimethylaminopyridine.

The yellow solution is stirred for 5 hours under anhydrous conditions at room temperature.

Then add 2-hydroxypropylamino (0.1 ml) and the mixture is stirred for 30 minutes.

The reaction mixture is neutralized 0.15 ml of glacial acetic acid, concentrated in vacuo to a small volume and transferred into a 150 ml acetone. The mixture is stirred for 1 hour, the precipitate is filtered off and washed with acetone receive of 5.85 g of the title compound.

The content of paclitaxel is 4% wt./wt.

Example 7.

2',7-Di(carbobenzoxy--alanyl)paclitaxel.

To a solution of 200 mg of paclitaxel in 15 ml of acetonitrile add 400 mg of N, N'-dicyclohexylcarbodiimide, 200 mg carboneria storytell evaporated to dryness in a vacuum.

Chromatographytandem residue on silica gel using as eluent a mixture of ethyl acetate - hexane (1 : 1) receive 300 mg of the title compound.

The Belarusian library Association-MS: m/z 1264/M + H/+, 1204, 1130, 1070.

Example 8.

7-(carbobenzoxy--alanyl)paclitaxel.

To a solution of 171 mg of 2',7-di(carbobenzoxy----alanyl)paclitaxel in 60 ml of methanol is added 30 mg of sodium bicarbonate and 7 ml of water. The reaction mixture is stirred 3 hours at room temperature.

The methanol is evaporated and the product extracted with ethyl acetate.

Evaporation of solvent to dryness in vacuo obtain 134 mg of the title compound.

Example 9.

7-(-alanyl)paclitaxel

To a solution of 135 mg of 7-(carbobenzoxy -- alanyl)paclitaxel in 20 ml of methanol and 13 ml of formic acid is added 200 mg of 5% Pd/C. the Reaction mixture is stirred 6 hours at room temperature. The catalyst is filtered off, washed with methanol and the solvent evaporated to dryness in a vacuum. The residue is dissolved in 8 ml of methanol and after deposition of 150 ml of diethyl ether to obtain 85 mg of the title compound.

The Belarusian library Association-MS: m/z 925/M + H/+, 947/M + 11a/+.

1H-NMR (400 MHz, CDCl3) : to 1.14 (s, 3H, CH3-16), 1,2 (s, 3H, CH3-17), to 1.79 (s, 3H, CH2CH2NH2), a 3.9 (d, J = 6,8 Hz, 1H, 3), 4,18, or 4.31 (two d, J = 8,2 Hz, 2H, CH2-20), and 4.8 (d, J = 8.5 Hz, 1H, 2'), 4,91 (d, J = 8.5 Hz, 1H, 5), 5,62 (LW. d, J = 10,2 Hz, J = 7 Hz, 1H, 7), to 5.66 (d, J = 6,8 Hz, 1H, 2), 5,81 (LW. d, J = 2,9 Hz, J = 9.1 Hz, 1H, 3'), 6,17 (m, 1H, 13), to 6.19 (s, 1H, 10), 7,3 - 8,2 (m, 16H, NH 4'+ 3-Ph).

Example 10.

Copolymer 1 methacryloylamido-2-hydroxypropane, 1-(methacryloyl-glycyl-i.e. phenylalanyl-leucyl-glycyl)amino-2-hydroxypropane and 7(methacryloyl-glycyl-i.e. phenylalanyl-leucyl-glycyl---alanyl)paclitaxel.

To a solution of 1500 mg copolymer 1 methacryloylamido-2 - hydroxypropane and N-(methylennorbornan-Phe-Leu-Gly-4 - nitrophenoxy)methacrylamide in 13 ml of anhydrous dimethylformamide added 135 mg of 7-(-alanyl)paclitaxel and 20 mg of dimethylaminopyridine. The yellow solution is stirred for 5 hours under anhydrous conditions at room temperature.

Then add 2-hydroxypropylamino (0.2 ml) and the mixture is stirred for 30 minutes.

The reaction mixture was neutralized with 0.3 ml of glacial acetic acid, concentrated in vacuo to a small volume and transferred into 250 ml of acetone. The mixture is stirred for 1 hour, the precipitate is filtered off and washed with acetone receive 1520 mg of the title compound.

The content of paclitaxel is 7.8% wt./wt.


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in which one of R1and R2represents the residue of a copolymer of the formula

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and the other is a hydrogen atom,

no more of 9.9 mol.% links formula

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where R is phenyl or tert-butoxypropan;

R3is hydrogen or acetyl;

A and a1that may be the same or different, represent simple chemical bond, amino acid residue or a peptide spacer selected from: - Ala, Gly, Phe-Gly, Phe-Phe, Leu-Gly, Val-Ala, Phe-Ala, Leu-Phe, Leu-Ala, Phe-Leu-Gly, Phe-Phe-Leu, Leu-Leu-Gly, Phe-Tyr-Ala, Phe-Gly-Phe, Phe-Phe-Gly, Phe-Leu-Gly-Phe, Gly-Phe-Leu-Gly-Phe, Gly--Ala, Phe-Gly--Ala, Phe-Phe--Ala, Leu-Gly--Ala, Val-Ala--Ala, Phe-Ala--Ala, Leu-Phe--Ala, Leu-Gly--Ala, Phe-Leu-Gly--Ala, Phe-Phe-Leu--Ala, Leu-Leu-Gly--Ala, Phe-Tyr-Ala--Ala, Phe-Gly-Phe--Ala, Phe-Phe-Gly--Ala, Phe-Leu-Gly-Phe--Ala, or Gly-Phe-Leu-Gly-Phe--Ala,

obtained by reacting the compounds of formula II

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where one of the A2and a3means a chemical bond and other means;

the values of R, R3and As mentioned above,

the activated polymer consisting essentially of 90 to 99.9 mol.% links formula

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and 10 to 0.1 mol.% links formula

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with the subsequent processing of the conjugate, which contains from 2.65 to 7.8 wt. % link formed from the active compounds of formula II, 2-hydroxypropylamino, and the process Escoge or inorganic bases.

2. Conjugate under item 1, in which R is phenyl, R3- acetyl, - Phe-Leu-Gly or Phe-Leu-Gly--Ala.

3. Conjugate under item 1 or 2, which is a copolymer of 1-methacryloylamido-2-hydroxypropane, (methacryloyloxy-i.e. phenylalanyl-leucyl-glycyl)-3-amino-2-hydroxypropane and 2'-(methacryloyloxy-i.e. phenylalanyl-leucyl-glycyl--alanyl)paclitaxel.

4. A method of obtaining a polymer conjugate under item 1, namely, that the compounds of formula II

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where one of the A2and a3represents a chemical bond, and the other is A, where A, R and R3take the values under item 1,

enter into interaction with the activated polymer as defined in paragraph 1, and the reaction is carried out in an anhydrous polar solvent at a temperature of 15 - 40oC for 1 to 24 h in the presence of organic or inorganic bases, and then processes the resulting conjugate 2-hydroxypropylamino.

5. The compound of formula II under item 1, in which R is phenyl, R3- acetyl AND2= Phe-Leu-Gly, Gly-Phe-Leu-Gly or Phe-Leu-Gly--Ala, AND2- a chemical bond.

6. The compounds of formula II under item 1, in which R is phenyl, R3- acetyl AND2- a chemical bond, AND3--Ala or Phe-Leu-Gly--Ala.

7. Pharmaceutical composition, oblast fact, as the current tools it contains the connection PP.1, 2 or 3 in an effective amount.

 

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The invention relates to aqueous drug composition having property of reversible temperature-controlled fermentation, which comprises a pharmacologically effective component, methylcellulose, citric acid and polyethylene glycol (PEG)

The invention relates to pharmaceutical industry and for the pharmaceutical tablets for the treatment or prevention of hypercholestremia and method of production thereof

The invention relates to medicine, namely to the composition and method of humidifying membrane tissue
The invention relates to methods for enterosorbent and allows you to get medical lignin in pill form
Ointment "linein" // 2076697
The invention relates to medicine, namely to the ointment "LINESIN used in dentistry for the treatment of periodontal disease

The invention relates to medicine, in particular to the technology of drugs, and may find application in the creation and production of finished dosage forms (powders, tablets), which include acetylsalicylic acid (AK)

, 20-epoxy-1,2, 4,7; , 1013th- hexahydroxy-11-en-9-it is complex ester group in position 13 formed (2r, 2s)-n-hexanoyl-3 - phenylisoxazol, method thereof and pharmaceutical composition" target="_blank">

The invention relates to a new taxane, namely 4,10-diacetate 2-benzoate 5, 20-epoxy-1,2, 4,7, 1013th-hexahydroxy-11-EN-9-it is complex ester group in position 13 formed (2R, 2S)-N-hexanoyl-3-phenylisoxazol having the structural formula I, which are extracted from the bark of the roots of plants of the genus Taxus extraction

The invention relates to the field of macrolides
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