RO109072B1 - Tesole derivates and preparation process therefor - Google Patents

Abstract

The invention relates to taxol derivatives and processes for their preparation, compounds we, represented by the general formula I: wherein R and R 'each represent hydrogen or the remainder of an amino acid selected from alanine, leucine, isoleucine, valine, phenylalanine, proline, lysine and arginine or a group of formula III: zRÎ - CO - [CHJ.- N <(III) o3 RO 109072 Bl wherein R and R 'each represent hydrogen or the remainder of an amino acid selected from alanine, leucine, isoleucine, valine, phenylalanine, proline, lysine and where n is an integer, from 1 to 3, and R2 and R 3 each represent hydrogen or a radical alkyl, with 1 ... 3 carbon atoms or together with the nitrogen atom, to which they are bound, forms a heterocyclic ring, saturated with 4 or 5 atoms carbon, provided that at least one of R and R ' not be hydrogen. Taxol derivatives have solubility in water, higher than taxol and can be formulated in pharmaceutical compositions, with a very good activity anti-tumor.

Description

ι 109072 2

The present invention relates to detaxol derivatives and processes for their preparation, compounds having improved water solubility compared to taxol and exhibiting very good antitumor activity, being particularly useful for the treatment of the same types of cancer in which its taxol proven active, including human lung tumors, melanoma, leukemia, breast tumors and colon cancer.

Taxol is a well-known dipterid, a sesquiterpene, which was initially isolated from the bark of the western stem strain. Toxus

Taxol has been found to possess potential antitumor activity against a wide variety of tumors. This compound, however, is very poorly soluble in water, which creates major problems in the development of appropriate pharyngeal pharmaceutical compositions useful in human therapy. Some taxol 15 compositions for intravenous injections or infusions were initially made using chromium-phore EI as a drug carrier to overcome problems related to low water solubility of taxol. But the creomophore itself is somewhat toxic, causing the idio-synchronic release of histamine and anaphylactoid reaction, so using this medium is not a desired solution to the problem of developing good taxol-based compositions.

Various taxol derivatives are known, used as anti-tumor agents, with the general formula:

RJO

1 OCOPh

The invention broadens the range of taxol derivatives with superior antitheor properties. with new compounds having the chemical structure corresponding to general formula I: wherein: R represents hydrogen or acetyl; R 'represents a PhCHR Cli-R'CO radical; JU represents hydrogen; one of RJ and R4 represents OH and the other an NHC02CMe3 radical (EP Appl. 253738). 4 109072

'iii which: R and R' represents each hydrogen or remainder of an amino acid selected from the group consisting of alanine, leucine, isoleucine, valine. phenylalanine. proline, isin and arginine saji a radical of the formula 5: / R &quot; CC) - (CH?) - N R ~ in which. n is an integer from 1 to 3 and R &quot; ' and R ~ represents each hydrogen or an alkyl radical with 1-3 carbon atoms, or R10 and Rvi together with the nitrogen atom to which they are attached form a hetero-cyclic ring, saturated with 4 or 5 carbon atoms. carbon, provided that at least one of R and R? not be hydrogen, or an acid addition salt thereof.

The compounds of the general formula I, according to the invention, are prepared by reaction of the taxol with a compound of the general formula II: 20 HO -? - (CH2) "- NR2R3 (II) wherein: &quot; R &quot; and R * 5 have the above meanings, in the presence of a condensing agent and, possibly, in the presence of a catalyst, forming a substituted compound, which undergoes a cleavage reaction.

Compounds of general formula I, wherein R and R 'represent each hydrogen or the remainder of an amino acid, provided that at least one of R and R' is not hydrogen, according to the invention, is prepared by reaction of the taxol with a 35 N amino acid -protected, after which the amino acid radical is deprotected.

Compounds of general formula I, wherein R is the residue of an amino acid selected from the group consisting of alanine, leucine, isoleucine, valine, phenylalanine, proline, isin and arginine, a radical of the formula: ^ ÎV ^ -CO - (CHz) -NR wherein w, IV and R have the abovementioned meanings according to the invention, are prepared by reaction of taxol with a compound protecting the hydroxyl group to form 2- (protein-jat) - taxol; its reaction with alanine, leucine, isoleucine, valine. phenylalanine, proline, lysine and arginine or with the general compound of formula II, followed by deprotection reaction.

The invention has the advantage of obtaining novel taxol derivatives which, formulated in pharmaceutical preparations, have superior cytostatic and antitomorphic activity, are water soluble and have good stability at the levels of pfl corresponding to the pharmaceutical compositions, but are rapidly destroyed in vivo. physiological pill, to act as a potential pro-taxol drug. Depending on the meanings of the substituents, the invention comprises: (a) esterified derivatives at the hydroxy group at position 2 'of the taxol, (b) esterified derivatives at the hydroxy group at position 7 of the taxol, and (c) esterified derivatives at the hydroxy groups at the positions 2 'and 7 of the taxol.

Among the compounds included in general formula I above, the following compounds 6 109072 may be considered preferred in the present invention: 1. 2M'N, N-diethylaminopropions) -taxol: 2 2 '- (N, N-dimethylglycyl) -taxol; 7- (N.N-dimethylglycyl) - taxol; 4 2 -7-di-N, N-d imetiIglieil) - taxol; £ 7- (N.N-diethylaminopropionyl) -taxol; 6.2 ^ -di- (N, N-diethylaminopropionyl) -ta- χοΐ: 7 2 '- (l-glycyl) -taxDl; &Lt; &gt; 7- (I.-li ci 1) -îaxo 1; g 2 -7-di- (L-glycyl) -taxol; To. 2 '- (L-alanyl) -taxol; jl. 7- (L-Alania) -taxol; 12. 2 '- (L-Alanyl) -taxol: 13. 2' - (L-leucyl) -taxol; 14. 7- (L-leucyl) -taxol; J5. 277-di- (L-leucine 1) -taxol: 16. 2 '- (L-isoleucyl) -taxol; J7. 7- (L-isoleucyl) -taxol; 18. 2? .7-di- (L-isoleucyl) -taxol; 19. 2 '- (L-valyl) -taxol: 20. 7- (L-vals) -taxol; 21. 27-di- (L -values 1) - taxo 1: 22. 2? - (L-phenylalanes 1) - taxol; 23.7- (L-phenylalanine methyl ester) -taxoi; 24. 2 \ 7-di- (L-phenylalanyl) -taxol; 25. 2 * - (L-prolyl) -taxol: 26. 7 - (L pro III) - taxol; 27. 2V7-di- (L-proiii) -taxol; 28. 2, - (L-lysyl) -taxol; 29.7- (L-lysyl) -taxol; 30. 2 \ 7-di- (L-lysyl) -taxol; 31. 2 '- (I -glutamyl) -taxol; 32.7- (L-glutamyl) -taxol; 33. 2 \ 7-di- (L-glutamyl) -taxol; 34.2 '- (L-arginyl) -taxol; 35. 7- (L-Arginyl) -taxol: 36. 2 \ 7 -di- (1-arginyl) -taxol. The chemical reactivity of the hydroxyl groups at positions 2 'and 7 15 of the taxol has been found to exist, the 2-hydroxy group being more chemically reactive than the 7-hydroxy group. This observation was then used to act in these positions and to create new derivatives of general formula I. The ester derivatives at position 2 'can be prepared by one of two methods, depending on whether the derivative will contain an amino acid or a amino acid acylate. For the preparation of their samples with alkylated amide-25, the reaction scheme is applied below;

Scheme I alkylated amino acid + taxol &gt; 2'-ester derivative of taxol

N

OH i c = o i &lt; CH2) / &gt; - I H-laxol

NT R- R 'where: n, above. For noae, the following: / V R2 and R3 have meanings preparation of ami esters apply the reaction scheme of 30

Scheme II N-protected amino acid + taxol derived from taxol-protected amino acid --► amino acid ester at the T position of taxol. 7 109072 8

In both of the above reaction schemes (Scheme I and Scheme II) the reaction of the alkylated or N-protected amino acid is carried out in the presence of a condensing agent, in the presence or absence of a catalyst, preferably at room temperature. With suitable condensing agents are carbodiimides. such as dicyclo-hexylcarbodiimide (DCC). Suitable catalysts are 4-dimethylaminopyridine 10 (DMAP) and pyridine. Various protecting groups for the amino group can be used in reaction scheme II and can also be used as known starting amino acid protected materials. Thus, 15 amino acids protected with fcf-butoxide (! -BOC), fluorenmethoxyoxide (FMOC) or carbobenzyloxy (CBZ) can be used. Amino acids protected with t-JBOC or FMOC are preferred.

Although deprotection of t-BOC group 20 of 2'-esters. Using aqueous formic acid solution and other organic acids, it leads to product degradation as well as stereochemical modification. better results were obtained using formic acid 25 09%. In the case of FMOC-protected amino acid esters, product recovery depends on the processing conditions. Thus, in the final deprotection step, the conditions used to remove the t-BOC protecting group may cause undesirable modifications to the free hydroxyl group at position 7 of the taxol molecules. These undesirable modifications consist of stereochemical modifications of the molecule. For example, N-protected alanine compounds may be represented as follows: CH3 9

CH3-C-0-C-NH-OH -COOH protected 40 CH3 CH3 t-BOC - CH 2-O-C-NH-CH-COOH protected CH3 FMOC 45

As it has been found the 2'-hi-droxyl group of the taxol is more chemically reactive than the 7-hydroxy group. So. In both schemes I and II above, the substitution or 50 ester is as bad occurs at position 2 'by reacting the alkylated or N- taxol protected amino acid, at a molar ratio of I: 1 or slightly greater than 1: 1. The reaction using equal molar amounts leads to the production of a large excess of the taxol derivative 2, although small amounts of taxol ester 7 may occur as a by-product.

Preparation of the esters of heading 7

Scheme III

Because the 2'-hydroxyl group of the taxol is more reactive than the 7-hydroxyl group, this esterification requires a process different from that used in the preparation of 29-derivatives. Thus, to prepare the esters from position 7, a process is used in which the 2'-hydroxy group is first protected or blocked, then the 7-hydroxy group is esterified and finally the protection or blocking group is removed from position 2. '*. taxol + protected compound - v 2'-pro-tiled taxol - &gt; with alkylated amino acid - &gt; taxol Z'-protected 7-acylamyl nonalkylated ^ Prot5Jar |. taxol 7- esterified.

A number of known blocking groups can be used to block position 2 of the taxol. Using an example of a blocking group and using an alkylated amino acid as an example, reaction scheme III can also be summarized as follows: taxol 4- 2,2,2-trichloroethyl chloroform t - &gt; 2 (barter) -taxol-fN.N - dimethylglycine 4- (barter) condensing agent -l-catalyst - * 2 '- (barter) -7- ί, Ν, Ν -dimethylglycyl) -taxol - -binding pos ^ 7_ (RN_ dimethylglycyl) -taxol. In the above reaction scheme, the reaction of 2 '- (barter) -taxol with the alkylated amino acid is carried out in the presence of a condensing agent and a catalyst. The corresponding condensing agents and catalysts are the same as those mentioned in Schemes I and II. Deprotection 2? - (trocHaxol can be carried out, for example, by using a mixture of zinc and acetic acid.

Scheme IV

As a variant of the process, 7-substituted taxol derivatives 109072 9 may also be prepared by a process in which the taxol first reacts with 2 ... 3 equivalents of N-blocked amino acid to produce taxol 2, - disubstituted, both amino acid groups at positions 2 'and 7. are deprocessed, then cleaved amino acid at position 2 \ This process can be represented as follows, using t-BOC-protected alanyl as the protected amino acid group: taxol + Nt -BOC-alanine --- &gt; 2'.7-di- '2 equivalents (L-alanyl) -taxol - -v? -E- &gt; 7- (L-alanyl) - taxol. In this process, similar to Scheme III, the reaction of taxol with the protected amino acid is carried out in the presence of a condensing agent and a catalyst. Deprotection of amino acid groups is accomplished by a known method of deprotecting amino acids, such as treating with a weak acid, for example formic acid. The cleavage of the 2-amino acid group is effected by bringing the pH of the solution of 2 \ 7- (amino acid) -taxol to 7-7,4, for example, by mixing 2 ', 7-di- (ami-noacid) - taxol with a phosphate buffer solution. IapH = 7 ... 7.4. PH correction in this way splits the 2'-amino acid group and the desired 7- (amino acid) -taxol is obtained. Thus, for example, taxol is treated with 2-3-molar equivalents of N-protected amino acid (t-BOC, CBZ or FMOC as protecting groups) in methylene chloride, in the presence of DCC and a catalytic amount of 4-dimethylaminopyridine. Thus, a protected amino acid group is introduced at position 2 'and at position 7. The protecting groups are removed with appropriate cleavage agents (eg, acid, weak base, hydrogenolysis). The 2 \ 7-di- (amino acid) derivative of taxol is kept in phosphate buffer solution at neutral hair for 24-48 hours, during which time the selective deprotection of position 2 \ takes place and the 7-substituted derivative is obtained of taxol.

A similar reaction scheme can be used to obtain the 7-substituted derivatives of taxol with alkylated amino acids 10. This reaction may be similar to the above, except that the N-protected amino acid group is replaced with a desired N-alkylated amino acid group and the deprotection step is eliminated. This scheme can be presented as follows:

Scheme V taxol-1-amino acid alkylated -7 &gt; 2 \ 7r (disuh-Established) -taxol -5Cinda-re- ":? Minoaad ^ cb, to taxol 7-substituted

Preparation of 2 \ 7-di-substituted taxol derivatives

The disubstituted derivatives can be prepared by using the above processes or part thereof. These procedures can be outlined as follows:

Scheme VI: substitution with alkylated amino acids: taxol + amino acid taxol 2-7 (disubsti- 2-3 equivalents tuite with alkylated amino acid).

Scheme VII: amino acid substitution: taxol + N-protected amino acid taxol 2V7- 2-3 equivalents (disubstituted c, one protected amino acid -? PP? - &quot; taxol 2 \ 7- (disubstituted with amino acid) 2-3 equivalents

Scheme VI is similar to Scheme I above, except that the reaction occurs with 2 equivalents of alkylated amino acid. In addition, although Scheme I may be carried out in the presence or absence of a catalyst, the process in Scheme VI requires the presence of a catalyst, due to the reduced chemical reactivity of the 7-hi-droxy group of taxol.

Scheme VII is similar to Scheme II above, but it is observed in Scheme II that FMOC is the preferred protective group in order to avoid stereochemical modifications of position 7, during deprotection. This problem is not encountered in reaction scheme VII because the 7-hydroxy group is not free. Thus, various known protection groups, including t-BOC and FMOC, can be used. 109072 11

If so. The following are examples of preparation of compounds of general formula I. according to the invention. 12

Example 1. Preparation of 2 '- (N, N-di-1-non-glycyl) -taxol or a salt thereof

3. r = o = C-CH? -N (CH3) 2 R '= H

2. R = 0 = C-CH2-N (CH3) 2.CH3S03H

R = H (a) Compound I. 2 (N, N-dimethylgU-cyclohexol) At a solution of 0.21 g (0.246 mmol) taxol and 0.0254 g (0.246 mmol) N, N-dimethylglycine in 12 mL methylene chloride year Hydra, 0.15 g (0.72 mmol) of 1.3 dicyclohexylcarbodiimide and O. 025 g (0.2 mmol) of 4-dimeiylaminophenyl are added.The reaction mixture is stirred under anhydrous conditions for one day. stirring is continued for another 6 hours. The reaction mixture is filtered and the filtrate is evaporated under nitrogen atmosphere The residue is chromatographed on silica gel (35 g, 26 cm) and eluted successively with ethyl acetate: 1: 1 petroleum ether and acetate. ethyl acetate: Fractions in ethyl acetate: petroleum ether after slow evaporation, give a filterable solid. The mother liquor is concentrated and petroleum ether is added until turbulence occurs and deposited to obtain more product. 0.14 g (61%) with a melting point between 168 and 171 ° C (decomposition).

In the NMR spectrum of compound 1 (300 MHz. CDCI3) the proton resonances at position 2 'change from 4.71 ppm to taxol to 5.59 ppm. This corresponds to esterification at position 2 '. All 5 other resonances of the spectrum are in accordance with the structure provided. The purity by high pressure layer chromatography (HPLC) is 98 ... 99.5%. Mass spectrum: (PAB) m / e 939 10 (Μ + H) +. Elemental analysis: calculated on n C51H58N2O1S: C = 65.26: H = 6.22: N = 2.98: found: C = 65.16: H = 6.2S: N = 3.13%. (b) Compound 2. Me-15 tansulfonic acid salt of 2W N, N-dimethylglycyl) - taro. 0.06 g (0.064 mmol) of 2 '- (N, N-dimethylglycyl) -taxol is dissolved in 2 ml of t-bu-tanol and 1.5 ml of water. The mixture was cooled to -5 ° C and 3.1 ml (2 mg / ml, 20 0.0645 mmol) methanesulionic acid was added. dropwise, and the mixture was stirred at 0-5 ° C for 1 min and filtered on a 20 µm millipore filter in a cooled flask in ice-isopropanol mixture. The solution was dried by freezing to give 0.058 g of product (80%), m.p. 170-117 ° C.

Elemental analysis: calculated for C52H02N2S18.2H2O: C = 57.83: H = 6.27: 30 N = 2.6: found: C = 57.49; H = 6.06: N = 2.73%.

Physical properties: molecular weight 1035; m.p. 170. „173 ° C (decomposition); solubility 15 mg / ml 13 109072 14 (slightly cloudy), 2 mg / ml (clear).

Chemical stability studies: the compound was subjected to stability studies according to the procedures described below. 5

The stability of the derivatives at different folds was studied at 25 and 37 ° C. Plasma studies were performed at 37 ° C on rat plasma and human plasma. The concentration used for the derivative was around 10 to 20 ... 25 pg / ml. A stock solution of compound of 0.8 ... 1.0 mg / ml was prepared and added to the plasma to give the desired concentration (20 ... 25 pg / ml). 100 µl samples were taken and stabilized with 250 µl 15 acetonitrile, centrifuged to precipitate plasma proteins. The degradation kinetics were studied by the peak of the HPLC diagram, namely by the area of this peak with respect to time. Tgo and 20 tso were calculated. Chemical and plasma studies were followed by high performance liquid chromatography (HPLC) using an RP-8 column (15 cm) and a precolumn (5 cm). The detector was set at 227 25 mm. The mobile phase consisted of 0.02 M acetate (pH = 5): acetonitrile 50:50 or 35:65 and the flow rate varied between 1 and 1.5 ml / min or the same solvents containing 0.001 M tetrabutylammonium acid sulfate with a flow rate of 1 ml / min. In stability studies, the disappearance of the peak of the compound led to the formation of a peak having a retention time equal to that of the taxol. The identity of this peak was further confirmed by the degradation studies of the new derivatives. Thus, 2 '(DMG) - taxol was incubated with water at 37 ° C and the product was concentrated and purified by preparative thin layer chromatography (TLC). After purification, the product was analyzed by HPLC and spectroscopic method. It has a molecular ionic peak at m / e 860 (Μ -I- Li) +, indicating that it is a taxol.

The working conditions at HPLC are: RP-8 column. length 150 mm, 4.6 mm id: mobile phase 0.02 M acetate (pi 1-5): acetonitrile 50:50: Kratos Spec-troflow 757 detector: flow rate 1 ml / min; retention time 11.2 ml at compound 2 and 5.5 nil at taxol. Chemical stability results: conditions ti / 2 (h); 0.02 M acetate (0.1 mg / ml) at pH = 3.5 and 25 ° C, 96.2. and at pH = 4.5 and 25 ° C 55.4; in water (2 mg / ml) at pH = 3.8 and 37 ° C. 89.8.

Plasma stability at 37 ° C. conditions 11/2 (min) - rat plasma (20 Mg / ml) 3.05 - canine plasma (20 Mg / ml) 121.6 - human plasma (20 Mg / ml) 198.6

Example 2. The 2'- (NM-diethyl-minopropionyl) - taxol salts

R '= H R' = H

R = 0 = C-CH2'CH2N (C2H5) 2.HC1 R β 0 = 6-CH2-CH2N (C2H5) 2.CH3S03H 109072 15 (a) Compound 3. HCl salt of 2- / 3- (N, A- di el Ham in o} - proion UI- taxoluluu To a solution of 0.12 g (0.14 ramol) taxol in 12 ml methylene chloride, containing 0.025 g (0.145 mmol) N, N acid hydrochloride -dietylaminopropionic 0.08 g (0.38 mmol) of 1,3-dicyclohexylcarbodii and 0.01 g (0.081 mmol) of 4-dime-tilaminopyridine were added. The resulting mixture was stirred at room temperature for 24 h. The reaction mixture was filtered and the filtrate was evaporated under a nitrogen atmosphere. The crude material was chromatographed on a silanized silica gel column (18 g, 22 cm) and eluted successively with ethyl acetate: petroleum ether 1: 2, ethyl acetate: ether 1: 1 petroleum and ethyl acetate Fractions In ethyl acetate: petroleum ether, by slow evaporation, it forms a solid precipitate which is filtered. The mother liquids containing the product are combined, concentrated and petroleum ether is added. turbulence and deposition The product is filtered and 0.068 g of product (48%) is obtained, with a mp of 188 ... 191 ° C, mass spectrum (FAR) rn / e 981 (M + H) +; NMR (300 Hz, CDCb) shows that the proton responses at position 2 'change from 4.71 ppm to taxol to 5.53 ppm. The ethyl groups have the methyl resonance 1.0 ppm and the CII2 resonance 2.52 ppm. All other resonance characteristics of the supposed compound were found. Elemental analysis for C51H6SCIN2O15: calculated: C = 63.73; 11 = 6.43; N, 2.75; found: 0 = 64.84; H = 6, S4; N = 2.89%.

Physical properties of compound 3: molecular weight 1017.56; mp 186 ... 189 ° C (decomposition); sol-lubricity -0.8 mg / ml. HPLC conditions: RP-8 column 150 mm long, 4.6 mm i.d .; mobile phase: 0.02 M acetate (pH = 5): acetonitrile! 35:65; Spectroflow 757 Root Detector; flow rate 1.5 ml / min; retained volume 16.71 ml (compound 3).

Plasma stability at 37 ° C: in human plasma 11/2 4.2 min. 16

Chemical stability: conditions t.1 / 2 (h) - 0.02M acetate (0.01 mg / ml, pill = 3.5; 25 ° C) 438.6 - 0.02M phosphate (0.02 mg / ml , pH = 7.4, 25 ° C) 0.25 (b) Compound 4. Methanesulfonic acid salt of 2- (A, A- diethylaminopropio-nyl) -taxate. To prepare the methanesulfonic acid salt of N, N-diethyl-aminopropionic acid, 10 g QAE-sepliadex (Pharmacia) is moistened with 0.1 M NaCl solution for 75 hours and 75% of this material is poured into a column. The column was washed with 700 ml of distilled water. Then the column was equilibrated with 500 ml CFGSOsNa (prepared from solutions of 0.5 M methanesulfonic acid and 0.5 M sodium hydroxide and titrated to pH = 6). The complete disappearance of ΟΓ was established by collecting the dye and testing for the chlorine ion by adding a few drops of 1% sodium acetate in a few drops of nitric acid. Then the column was washed with distilled water to your neutral pH.

In the column, pour 2.5 g of N, N-diethylaminopropionic acid hydrochloride dissolved in 15 ml of water and elute with water. collect 4 fractions of 50 ml each. The first few fractions contain the product. These fractions are combined and the solvent is removed. The residue was dissolved in methylene chloride ethanol mixture and dried over magnesium sulphate and the solvent was removed to give 3.1 g of product. It is precipitated from ethanol and ether to give 2.6 g salt with methanesulfonic acid of N, N-diethylaminopropionic acid.

To a solution of 0.05 g (0.058 mmol) taxol and 0.014 g salt with methanesulfonic acid N, N-diethylaminopropionic acid (0.058 mmol) in 10 ml methylene chloride, add 0.061 g (0.3 mmol) 1.3 - dicyclohexycarbodiimide. The mixture was stirred at room temperature for 24 h. The reaction mixture was filtered and the filtrate was evaporated under a nitrogen atmosphere. The residue was chromatographed on a silanized silica gel column and eluted 109072 17 with 1: 1 ethyl acetate: petroleum ether and ethyl acetate. Fractions in ethyl acetate: petroleum ether by slow evaporation give 0.048 g product (74%), m.p. 170-14.4 ° C. Its physical properties are: molecular weight 1077.12; mp 170 ... 174 ° C; solubility &gt; 10 mg / ml; HPLC purity &gt; 99%. Chemical stability: conditions tt / 2 (h) -acetate 0.02M at pH = 4.5 and 25 ° C 305 -acetate 0.02M at / gt; H = 5.5 and 2S ° C 20.7 Example 3. Preparation 7- (N, N-dime-tert-D-taxol or a salt thereof) Compounds of the general formula indicated in Examples 1 and 2 are prepared, wherein R and R 'represent:

5. R = t) = C'-0-CH: &lt; x: i3 RH 6. RO ^ .- 0-CH2-CC13 R '* 0 = C-CH2-N (CH3) 2 7. RH R' = -OC-CH2-N (CH3) 2

8. R = H R'AC-CH2-NCCH3) 2.CH3SO3H (a) 2 '- (barter) -taxol (Compound 5.) Dissolve 0.27 g (0.316 mmol) taxol in 10 ml of methylene chloride and 1.5 ml pyridine. The reaction mixture is cooled to -20 to -25 ° C and 80 μ \ 2,2,2-trichloro-ethylchloroformate is added. The reaction mixture was stirred at this temperature for 3 hours. A further 25 μl chloroformate was added and the reaction mixture was stirred overnight. The reaction mixture was diluted with 50 ml of methylene chloride and washed successively with 25 ml x 2 HCl (), 1N and 25 ml x 1 NalICG3 0.05 M, cold and water. The organic extract was dried over anhydrous magnesium sulfate and the solvent was removed. The crude material is purified by preparative thin layer chromatography on silanised silica gel plates and removed in ethyl acetate: petroleum ether 1: 3, a portion is removed over the taxol, eluted with ethyl acetate and the solvent removed, yielding 0.32 g (97%) of mp 221 ... 226 ° C (decomposition, softening 160 ° C). (b) 2- (barter) -7- (A (N-dimethylglycyl) ~ taxai (Compound 6.) A mixture of 0.27 g (0.262 mmol) 2 '(barter) -taxol and 0.054 g (0.524 mmol) Ν, Ν-dimethylglycine is dissolved in 15 ml of methylene chloride, 0.215 g (1.04 mmol) 1,3-dicyclohexylcarbodiimide and 0.025 g (0.2 mmol) 4-dimethylaminopyridine are added to this solution. The mixture was stirred at room temperature for 2 days The reaction mixture was filtered and the solvent was removed The crude material was purified by preparative TLC chromatography on silica gel plates and extracted with 1: 1 ethyl acetate: petroleum ether. Remove a portion of the taxol (Rf 0.47, ethyl acetate: 1: 1 petroleum ether) and elute with ethyl acetate, the solvent is removed and 0.26 g of product (89%) is obtained with a mp 176-180 ° C (decomposition) Elemental analysis: calculated for C54H6OCI3N2O17: C = 58.11; H = 5.42; N = 2.51; found: C = 58.68; H = 6 , 00: N = 3.18%. (C) 7- (N, N-dimethylglycyl) -axol (Compound 7) To a solution of 0.335 g (0.3 mmol) 2 * - (barter) -7- (N, N-dimethylglycyl) -ta-xol in 12 ml methanol-9: 1 acetic acid is added 0.275 g powder of zinc and the mixture was stirred at room temperature for 25 minutes and filtered. The filtrate is concentrated to about 1 ml and diluted with 35 ml of methylene chloride and washed successively with 20 ml x 2 HC10.0IM, cold sodium hydrogen carbonate and water. The organic extract was dried over anhydrous sodium sulfate and the solvent removed to give 0.24 g of product. This compound was purified by preparative TLC chromatography on silanised silica gel plates (20 x 20, 3Nos) and removed in 7: 1 methylene chloride acetate chloride. The corresponding band 7- (DMG) of the taxol (Rf 0.35) is cut and eluted with ethyl acetate and ethanol, the solvent is removed and 0.19 g of product (68%) is obtained with melting point 18G ... 185 ° C (decomposition, softening to about 140 ° C). Mass spectrum (FAB) m / e 939 (M + H) +. In the NMR spectrum (300 MHz, CDCl 3) the hydrogen resonances from position 7 at 4.33 ppm in the taxol appear as double to the double at 5.65 ppm. The resonance of -N (CH3) 2 group appears as a single 109072 19 at 2.35 ppm. The methylene group of glycine is found at 3.16 ppm. (d) Methanesulfonic acid salt of 7- (dimethylglycyl) - taxol (Compound 8). Dissolve 0.065 g (0.069 mole) 7- (dimethylglycyl) -taxol in 2.5 ml and 1 ml water. The solution was cooled to 5-10 ° C and 3.36 ml (2 mg / ml, 0.0697 mmol) methanesulfonic acid was added. The mixture was stirred for 2 min and filtered through the millipore filter, in an ice-cold flask. The filtrate is freeze-dried and 0.066 g of product (94%) is obtained, with mp 164 ... 168 ° C (decomposition). Elemental analysis: calculated for: C52H62N2O18S. 2H2O: C, 58.29; H, 6.19; N, 2.6; found: 0 = 58.05; H, 6.00; N = l, 72.

Physical properties: molecular weight 1035; mp 164 ... 168 ° C; solubility &gt; 2 mg / ml. HPLC conditions: RP-8 column length 150 mm, 4.6 inner diameter; mobile phase acetate 0. 02M (/? H = 5): acetonitrile 35:65; Kratos Spectroflow 757 detector; flow rate 1.5 ml / min; retained volume 15.07 ml. (Compound 8) Chemical stability: conditions ti / 2 (h) - 0.02M acetate; / &Gt; H, 3.5; 25 ° C 3397 -acetate 0.02M; pH = 4.5; 25 ° C 1719 - 0.02M acetate; / &Gt; H, 7.4; 25 ° C 33.8

Plasma stability at 37 ° C: conditions ti / 2 (h) - rat plasma (20 ^ g / ml) 17.3 - human plasma (20 // g / ml) 27.7 - human plasma (10 / * g / ml) 24.4

Example 4. Preparation of 2 7- (dimethyl-glycyl) -taxol. The structure of the compound is that of the general formula of Example 1, wherein, R = R, 0 &lt; - OH 2 -N (NCH 3) 2 (compound 9).

Dissolve 0.06 g (0.0702 mmol) taxol in 5 ml anhydrous methylene chloride and add 0.015 g (0.145 mmol) N, N-di-methylglycine. To this mixture was added 0.08 g (0.368 mmol) 1,3-dicyclo-hexylcarbodiimide and 0.008 g (0.065 mmol) 4-dimethylaminopyridine. The reaction mixture was stirred at room temperature for 24 h and filtered. The solvent is removed from the filtrate. The residue is purified by preparative TLC on silanised silica gel plates and removed with ethyl acetate: 1: 1 petroleum ether. A band (Rf 0.17) was cut and removed over dimethylaminopyridine and eluted with a mixture of ethyl acetate and ethanol, and the solvent was removed. The residue is recrystallized from ethyl acetate: petroleum ether to give 0.046 g of product (64%) with mp 194 ... 198 ° C. Mass spectrum m / e 1024 (M ~). In the NMR spectrum (300 MHz, CDCl3) proton C2 (4.71 ppm) and proton C7 (4.33 ppm) changed to 5.5 ppm and 5.6 ppm respectively, demonstrating esterification at positions 2 'and 7. Also , the N-CH3 proton appears as a single at 2.3 ppm.

Elemental analysis: calculated for: θ55ΐΙόόΝ2Οΐό: C = 63.39; II = 6.48; N, 4.03; found: C, 63.00; H, 6.98; N, 3.98.

Synthesis of the 2'7- (dimethylglycyl) - taxol dimetansulfonic acid salt. To a solution of 60 mg (0.0585 mmol) 2.7- (dimethylglycyl) -taxol in 2 ml butanol and 1 ml cooled water at 0-5 ° C was added 11.37 mg (3.79 ml, 0.117 mmol) methanesulfonic acid. The α-mixture is stirred at 0-5 ° C for 2 minutes and filtered through the millipore filter (0.2μΜ), the filtrate is dried by cooling and 62 mg of melting point are obtained. ° C. Physical properties: molecular weight 1217; mp 160.163 ° C (decomposition); solubility &gt; 10 mg / ml; HPLC purity about 96%. HPLC working conditions: RP-8 column 150 mm long, inner diameter 4.6 mm; mobile phase acetate 0.02M (pH = 5); acetonitrile 60:50, containing 0.005 Μ TBA; Kratos Spectroflow 757 detector; flow rate 1 ml / min; retained volume 6.64 ml; 5.8 ml of taxol.

Example 5. Preparation of 7- (L-alanyl) ~ taxol or its (a) 2 \ 7-Di- (t-BOC-alumni) -taxol salt. To a solution of 0.21 g (0.246 mmol) taxol and 0.14 g (0.739 mmol) Nt-BOC-L-alanine 109072 21 in 15 ml methylene gold add 0.25 g (1.21 mmol) 1 , 3- dicyclohexylcarbodyne. The mixture was stirred at room temperature for 24 h and filtered. The residue was chromatographed on silica gel column (20 g, 14 cm) and eluted with ethyl acetate: 1: 1 petroleum ether and ethyl acetate. The fractions in petroleum ethyl acetate containing the substituted derivative are also combined

The solvent is removed to give 0.27 g compound (92%), m.p. 158-116 ° C (decomposition). (b) 7- (L-Alanyl) -taxol. Mix 0.29g (0.242 mmol) 2,7-di- (t-BOC-L-ala-nyl) -taxol and 2 ml formic acid and stir at room temperature for 40 minutes and excess formic acid is added. remove under nitrogen atmosphere The residue is dissolved in ethanol and petroleum ether is added. The solid was filtered to give 0.27 g 2 ', 7-di- (alanyl) -taxol. The crude dialanyl derivative thus obtained is taken up in 4 ml of acetonitrile! and 50 ml of phosphate buffer solution (0.02M, / day 1 = 7.4) and the mixture was stirred at room temperature for 12 hours. Increase the pH of the solution to 6.8 using several ml of amino acid phosphate. 5% sodium. The unclear solution is stirred at room temperature for another 8 hours. The reaction mixture is diluted with 50 ml of methylene chloride and 50 ml of cold 0.05 M sodium hydrogen carbonate solution is added. The reaction mixture was immediately extracted with 50 ml x 3 methylene chloride and the organic extract was washed once with water and dried over anhydrous sodium sulfate. The solvent was removed and 0.24 g of product was obtained. This compound is purified by silica gel column chromatography on silane gel giving 0.135 g of product (63%); purity &gt;95%; mp 159 ... 163 ° C.

Mass spectrum (FAP) m / e 925 (M + H) +. In the NMR spectrum (300 MHz, CDCl3) the hydrogen at position 7 in the taxol at 4.33 ppm appears as a doublet of the doublet at 5.65 ppm. The CH3 group on the alanine radical appears as a double at 1.27 ppm.

Elemental analysis: calculated for: C50H58N2O10. 2.5 H2O: C, 61.91; 22H - 6.54; N, 2.89; found: C, 61.41; H, 6.59; N = 2.78%. (c) The methanesulfonic acid salt of 7- (moles) - taro iuli. a solution of 62 mg (0.0658 mmol) 7- (alloy) taxol in 2 ml Jerf-butanol and 1 ml water cooled to 0. ..5 ° C 6.39 mg (2.13 ml, 3 mg / ml) methanesulfonic acid was added and the mixture was stirred at this temperature for 2 min and filtered through millipore filter (0.2 μΜ). The filtrate was dried by freezing to give 66 mg of product, with a melting point of 180 ... 184 ° C.

Physical properties: molecular weight 1021; mp 180 ... 184 ° C (decomposition); solubility &gt; 2 mg / ml. IIPLC working conditions are: RP-8 column with a length of 150 mm, internal diameter 4.6 mm; the mobile phase is 0.02M acetate (/? H = 5): 50:50 acetonitrile, containing 0.001 M tetrabutylammonium acid sulfate; Kratos Spectroflow 757 detector; flow rate 1 ml / min: volume retained 8.7 ml versus 7.1 at taxol. Plasma stability at 37 ° C, that is, in human plasma (20 / zg / ml) at 1/2 is 11.9 h.

Example 6. Preparation of 2 '- (alanyl) -taxol (a) Synthesis of 2' - (CBZ-L-alanyl) -taxol. To a solution of 30 mg taxol (0.036 mmol) and 8.5 mg (0.036 mmol) CBZ-L-alanyl in 5 ml methylene chloride was added 45 mg DCC and 4 mg 4-dimethylaminopyridine and the reaction mixture stirred at room temperature. , for 3 days. The reaction mixture was filtered and the solvent was removed from the filtrate. The residue was purified by preparative TLC on silanised silica gel plates and developed in 1: 1 petroleum ethyl ether acetate. The band above the taxol is cut and eluted with ethyl acetate and the solvent removed to give 28 mg 2 '- (CBZ L-alanyl) taxol. (b) Synthesis of 2 - (hazelnuts) -taxol by deprotecting 2 * - (CBZ-L- hazelnuts) -taxol. Dissolve 2 * - (CBZ-L-alanyl) -taxol in ethanol in the presence of an organic acid such as acetic or formic acid and stir at room temperature for 2 hours in 24 109072 23 in the presence of 5% palladium on coal. The reaction mixture was filtered to remove the catalyst, and the solvent was also removed. The crude product was dissolved in ethanol and petroleum ether 5 was added to give 2, - (alanyl) -taxol as formate or acetate, in low to moderate yield.

EXAMPLE 7 Preparation of 2 '- (lizU) -to-shampoo] t': .. »--- ru. &Lt; 'Λΐ_ &gt;' - &gt; u_t is cut, eluted with methylene chloride, the solvent removed to give 48 mg product which is 2 '- (FMOC-L-ala-nyl) -taxol, with mp 162 ... 164 ° C. (b) Deprotection of the FMOC group of the protected amino acid taxa derivative. The N-FMOC-protected amino acid taxol derivative reacts with piperidine in methylene clonir for 2 hours and the solvent is removed. The residue is 109072 25 it.

The new compounds are administered in the form of tablets, pills, powder mixtures, capsules, injections, solutions, suppositories, emulsions, dispersions, premixtures in food and other suitable forms.

The pharmaceutical preparation containing the compound is conveniently mixed with an organic, non-toxic pharmaceutical carrier or an inorganic, non-pharmaceutical pharmaceutical material carrier, typically about 0.01 mg to 2500 mg, or more per dosage unit, preferably 50. ..500 mg.

Amongst the pharmaceutically acceptable carrier materials are, for example, ma-nitol, urea, dextran, lactose, cauliflower and maize starch, magnesium starch, talc, vegetable oils, polyalkylene glycols, ethyl cellulose, poly (vi-nylpyrrolidone) ), calcium carbonate, ethyl oleate, isopropylmiristate, benzylbenzoate, sodium carbonate, gelatin, potassium carbonate, silicic acid and other acceptable carriers, conventionally used. The pharmaceutical preparation may also contain non-toxic auxiliary substances, such as emulsifiers, preservatives, humectants, and the like, for example, sorbitan monolaurate, trieta-nolaminoleate, polyoxyethylene monostearate, tripe Glyceryl imitate, di-ethylsulfosuccinate or dioctylsodinate also.

As an example of a typical method of preparing a tablet containing active agents, it is first of all to mix the agent with a non-toxic binder, such as gelatin, acacia mucilage, ethyl cellulose and so on. The mixing is performed well in a standard type mixer and usually under anhydrous conditions. Then, the prepared mixture can be passed through conventional tablet machines where they form bars that turn into tablets. Freshly prepared tablets may be covered or left uncovered. Suitable, representative coating materials are non-toxic coatings such as shellac. methylcellulose, wax wax, maleic acid styrene copolymers, etc. For 26 oral administration tablet tablets containing 0.01, 5, 25, 50, 500 mg, etc., up to 2500 mg are manufactured, according to the above and based on the manufacturing methods well known in the art.

To form a tablet, mix the active compound, wheat starch, lactose, dicalcium phosphate and calcium carbonate evenly under anhydrous conditions, in a conventional mixer, until all the ingredients have mixed evenly. Then, a wheat starch paste is prepared as a 10% paste and combined with the freshly prepared mixture until a uniform mixture is obtained. Then, the mixture is passed through a standard sieve with few rnesli, dried in an anhydrous atmosphere and mixed with calcium stearate, compressed into tablets and coated, if desired. Other tablets containing 10.50, 100, 150 mg, etc., are prepared in the same way. The following composition is an example of a tablet formulation containing a compound according to the present invention.

Composition /. It consists of a tablet of 50.0 mg active compound, 15.0 mg wheat starch, 4.5 mg wheat starch paste, 15.0 mg calcium carbonate, 67.0 mg lactose, 2.0 mg stearate calcium and 50.0 mg dicalcium phosphate.

The manufacture of capsules containing between 10 mg and 2500 mg for oral administration consists mainly of mixing the active compound with a non-toxic carrier and closing the mixture in a polymer package, usually gelatin or the like. Soft capsules can be manufactured by incorporating the compound as an intimate dispersion into an edible, compatible carrier material, or strong capsules can be made, consisting essentially of the new compound mixed with a non-toxic solid, such as talc, stearate. calcium, calcium carbonate etc. Capsules containing 25, 75, 125 mg, etc., of the novel compound, alone or a mixture of two or more novel compounds, are prepared, for example, as follows. 109 072

DAY

Compozifa 11. 50.0 mg of active compound, 100.0 mg of calcium carbonate, 200.0 mg of lactose, 130.0 mg of starch and 4.5 mg of magnesium stearate are mixed together in a standard capsule. The mixture is discharged into a standard capsule. When we want to increase the concentrations of the active agent, the amount of lactose is reduced accordingly.

The compounds of the present invention should be freeze-dried and, if desired, combined with other pharmaceutically acceptable excipients to prepare injectable parenteral compositions. For this administration, the composition may be reconstituted in water (normal, saline) or in mixture of water and organic solvent, such as propylene glycol, ethanol, etc.

The dose administered, whether it is a single dose, multiple doses or a daily dose, varies of course depending on the particular compound chosen from those according to the invention, due to the variation of the potency of the compound, the route of administration chosen, the size of the recipient and the dose the patient's condition. The dose to be administered is not subject to limitations, but will usually be an effective amount or equivalent of a molar base of the pharmacologically active free form produced from a dosage formulation during the metabolic release of the active drug to achieve its desired pharmacological and physiological. The dose administered is generally in the range of 0.8 to 8 rng / kg body weight or about 50 ... 275 mg / n of the patient, especially about 230 ... 275 mg / m.

Biological activity. As shown above, the taxol derivatives of the present invention are useful because of their antitumor activity. The compounds are particularly useful for treating the same forms of cancer in which taxol has been found to be active, including human lung tumors, melanoma, leukemia, breast tumors, and colon cancer.

The biological activity of taxol derivatives was tested by (A) in vitro studies, 28 to measure the kinetics of the microtubular assembly and (B) in vitro to study the kinetics of B16 melanoma cell culture, as well as in vivo MX-1 pexenograft studies. adrenal capsule SRC. (A) In vitro kinetics of microtubular assembly kinetics. Microtubules are an integral part of eukaryotic cells and the microtubular assembly is of importance in relation to cell division and multiplication. It has been shown that microtubule polymerization is highly sensitive to calcium; calcium may inhibit tubulin assembly (binding) and depolymerize the preassampled microtubules. Known antitumor compounds have been studied for their effect on microtubule binding.

Vinca alkaloids, such as vin-blastine and vincristine, have been shown to break down cell microtubules, that is, in vitro have been shown to inhibit microtubule assembly and depolymerize solidly bound microtubules. Similarly, colchina has been shown to depolymerize microtubules in cells.

On the other hand, taxol has been shown to possess a unique mechanism of action, in that it promotes microtubule binding, but inhibits their disassembly, thus intervening in the G2 and M phases of cell cycle and division. In vitro studies have shown that microtubules, once polymerized, resist the presence of taxol upon depolymerization by other agents, such as calcium chloride or low temperature, which normally depolymerizes microtubules.

The studies were conducted to investigate the effect of taxol derivatives on microtubule binding. The microtubule assembly study was performed using both 2 'and 7 derivatives, according to in vitro procedures known in the literature (Mellado et al., Biochemical and Biophysical Research Communications, vol. 124, no.2 (1984), pp. 329-336; Magri et al., J. Org. Chem. 51, 797-802 (1986) and Pamess et al., Biochemical and Biophysical Re 109072 29 search Communications, vol. 105, No. 3, pp. 1082 ... 1091 (1982) The ease with which these compounds bind the microtubules is in the following order: taxol 7- (N, N-dimethylglycyl) taxol (11) 2'- (NN-diethylaminopropionyl) - taxol (8) 2 '- (N, N-dimethylglycyl) - taxol (2).

This study shows that a 2 'hydroxy group is essential for microtubule binding. The 2 'derivatives are only active if the 2'-hydroxy group is left free during the experiment. The derivatives 7- have free 2'-hydroxyl and are therefore active. This result is consistent with the 2'- and 7-fold activities of taxol known in the literature. (B) In vitro culture studies of Bl 6 cells. To confirm the activity of the taxol derivatives of the present invention, in vitro cell culture studies were performed using B-16 melanoma cells. These studies were performed according to standard methods (Donoso and Himes, Cancer Biochem. Biophysics, vol. 7, p. 133 (1984). In the study of B-16 melanoma cell proliferation, the order of efficiency is as follows: taxol 2 * - (N, N-dimethylglycyl) -taxol 2 '- (N, N-diethylamino-propionyl) -taxol 7- (N, N-dimethylglycyl) - taxol. This study and other kinetic studies show that 2-derivatives return to taxol and therefore, 2 * · derivatives are likely to act as a pro-drug. On the other hand, 7-derivatives have their activity as such and do not act as a drug aid. (C) In vivo studies. experience to confirm the biological activity of taxol derivatives includes in vivo studies in mice with subcutaneous capsule of human breast carcinoma MX-1 xenograft (NIH Publication no. 84 ... 2635, In Vivo Cancer Model, 1984, p. 23 In this test, groups of tested mice (6 mice per group) are used and g is used. control breaks of 12 mice. A tumor fragment (Whose human breast MX-1 xenograft) is implanted under the membrane covering the kidney of each mouse. The following test scheme is followed:

Day 0: The animals are anesthetized. Note the body weight (day 1 weight). The tumor is implanted, measured and noted. The animals are analyzed by accident after recovering from anesthesia. Bacterial cultures grow. The solubilities of the test agent are determined. Death is noted daily.

Day 1: Crops are checked. The experiment is abandoned if it has been contaminated. Prepare the test materials, make injections with the test agent (in the neck) based on the individual body weight. Treatment is Q4D on days 1.5 and 9. A fresh test agent is prepared for each injection day and administered based on the individual body weight of that day.

Day 2: The crops are reverified. It is tested discontinuously if they have been contaminated and noted according to the test.

Days 5 and 9: Fresh test agent is prepared for each day of injection and administered based on the individual body weight of that day.

Day 11: The experiment is completed and evaluated. We note body weights (weight on day 2). The tumor is measured in the OMU (Ocular Micrometer Unit - 10 OMU = 1 mm) and noted.

Evaluation. The measured parameter is the average change in tumor weight (delta) based on the length and width of the measurements in millimeters. The average body weights are noted for day 1 and day 11, T / C is calculated for all groups tested with over 65% survivors on day 11. Excessive difference in body weight change can be used in the toxicity assessment (test control minus). The dimensions are measured and noted in OMU units. A calculator is used for the following: 1) transformation of OMU units into mm: 2) calculation of tumor weights (mg) from tumor dimensions (mm x 109072 31 mm) based on the formula for the volume of a flat ellipsoid: I. X W2 2 in which L is the length of the 2 measurements; 3) we calculate the change (delta) of the average tumor weight for each group of mice, namely the change of the average tumor weight = the final average weight of the tumor - the initial average weight of the tumor; 4) the change (delta) of the average tumor weight for the test (T) and control (C) groups is calculated; 5) calculate T / C% for all test groups with over 65% survivors on the day of the final evaluation:

AWtT T / C% = - xlOO - if AWtT is positive

AWtC 32

AWtT T / C% = - xlOO - if AEtT is negative initial mean tumor weight test 5 Activity criteria. A T / C &lt; 20% is considered necessary to demonstrate moderate activity. The T / C value of 10% is considered a significant activity. The results of this test performed 10 for some taxol derivatives are presented in tables 1 and 2 where the tests are performed on human breast carcinoma.

From these results it can be seen that the taxol derivatives of the present invention 15 have very good antitumor activity. Therefore, these compounds are useful as antitumor agents based on their biological activity and their increased solubility in water, relative to taxol. 20

Table 1

Tests for human breast carcinoma

Treatment Results - Average tumor weights Nr. Compound Dose T / C Eroa Eroa Change # of group NSC (mg / kg) MGS% re MGS live weight regimen AB useful A calculation / total 0001 control 0.000 CNTL 0.75 0.1 12.79 2.4 12 , 04 11/12 0102 taxol 22.40 TOXb 0.60 0.1 0.00 0.0 3/6 -2.6 -0.9 0103 ?! 15.00 CRc 0.81 0.1 0.00 0.0 -0.81 6/7 -4.2 -2.5 0104 ?! 10.00 -84 0.58 0.1 0.09 0.1 -0.49 6/6 -2.6 -0.3 0105 ?! 6.67 CR 0.74 0.1 0.00 0.0 -0.74 5/5 -1.5 0.2 0106 ?! 4.45 CR 0.79 0.1 0.00 0.0 -0.79 5/5 -1.2 0.5 0207 at 30.00 TOX 0.68 0.1 0.00 1/6 -3.8 -2 , 1 0208 1! 20.00 CR 0.56 0.1 0.00 0.0 -0.56 4/6 -2.2 -0.5 0209 ?! 13.30 CR 0.64 0.1 0.00 0.0 -0.64 4/6 -2.3 -0.6 0210 f! 8,90 -53 0,77 0,1 0,36 0,4 -0,41 5/6 -0,9 0,8 0211 H 5,90 -78 0,61 -Oii - 0.13 oq ..... - 0.48 6/6 -2.2 -0.5 - a = compound 1 = 2 '- (N, N-dimethylglycyl) -taxol; - b = TOX indicates the level of toxicity; - c = CR indicates complete remission of the tumor; - A = weight change of the body tested; - B - the difference in weight of the body of the animal. 109072 33 34

Table 2

Tests for human breast carcinoma

Treatment Results Average tumor weights Nr. Compound Dose T / C. Eroa Eroa Change # of group animals NSC (mg / kg)% MGS re MGS re weighted in life AB useful Λ calculated / total 0310 taxol 15.00 TOXC 0.80 0.1 0/5 0311 ff 7.50 ΊΌΧ 0.82 0.1 0.00 0.0 3/5 -1.7 2.3 0312 Pf 3.75 CR 0.71 0.1 0.1 0.0 0.0 -0.71 4/5 -2.9 1.1 0313 ff 1.88 5 0.80 0.1 1.51 1.51 0.71 4/5 -1.0 3.0 0001 control 0.00 CNTL 0.77 0.1 15.38 2.2 14.61 10/11 0102 7a 33 , 00 CR 0.74 0.1 0.00 0.0 -0.74 4/5 -2.7 13 0103 ff 16.50 -93 0.59 0.1 0.04 0.0 -0.55 3/4 -5.7 - 1.7 0104 f »8.25 23 0.81 0.1 4.19 1.5 3.38 4/4 -3.9 0.1 0105 ff 4/13 34 0.74 0.1 5.66 1.3 4.92 4 / 5 -4.5 -0.5 0206 3b 36.00 TOX 0.75 0.1 0/5 0207 ff 18.00 CR 0.82 0.1 0.00 0.0 -0, S2 3/4 -3.3 0.7 0208 ff 9.00 CR 0.87 0.1 0.00 0.0 -0.87 5/5 -2.7 1.3 0209 ff 4.50 CR 0.78 0.1 0.00 0.0 -0.78 5 / 5 -2,8 1.2 - a = 7- (N, N-dimethylglycyl) -taxol; - b = 2, - (3- (N.N-diethylarylino) propionyl) -taxol hydrochloride; - c = ΊΌΧ indicates the toxic level; - d = CR indicates complete remission of the tumor; - A = weight change of the body tested; - B = the difference in weight of the animal's body.

Claims 1. Derived from taxol, characterized in that they have the structure corresponding to general formula I:

109072 35 wherein: R and R 'represent each hydrogen or remainder of an amino acid selected from the group consisting of: alanine, leucine, isoleucine, valine, fentanyl, proline, lysine and arginine or a group of the formula: -C0- (CH2V- N-R &quot; R3 wherein: n ^ is an integer from 1 to 3 and R ^ and R3 are each hydrogen or an alkyl radical with 1 to 3 carbon atoms or R ^ and R3 together with the nitrogen atom to which they are linked together form a heterocyclic ring saturated with 4 or 5 carbon atoms, provided that at least one of R and R 'is hydrogen or an acid addition salt thereof, A compound according to claim 1, characterized in that at least one of R and R 'is selected from the group consisting of L-allyl, L-leucyl, L-isoleucyl, L-vals. L-phenylalanyl, L-propyl, L-lysyl, L-arginyl. A compound according to claim 1, characterized in that at least one of R and R 'is an eruption of the formula: -CO- (CH2) -N-R2' R3 in re: is an integer from 1 to 3 and R &quot; and R 3 are each an alkyl radical with 1-3 carbon atoms. A compound according to claim 1, characterized in that it is 2 '- (N, N-dimethylglycyl) -taxol or an acid addition salt thereof. 5. A compound according to claim 1, characterized in that it is 2 '- (N, N-diethylaminopropionyl) -taxol or an acid addition salt thereof. A compound according to claim 1, characterized in that it is 7- (N, N-dimethylglycyl) -taxol or an acid addition salt thereof. 7. A compound according to claim 1, characterized in that it is 2 ', 7-di- (Ν, Ν-dimethylglycyl) -taxol or an acid addition salt thereof. 8. A compound according to claim 1, characterized in that it is 7- (wool) -taxol or an acid addition salt thereof. 9. Taxol derivatives according to claim 1, characterized in that together with the pharmaceutically acceptable carrier materials, diluents or excipients, they form pharmaceutical compositions for use in the treatment of tumors, such as human lung tumors, melanoma, leukemia, mammary tumors. and colon cancer. 10. Process for the preparation of taxol derivatives of general formula I, characterized in that the taxol is treated with a compound of general formula II: wherein: w, R "and R ~ have the above meanings. 11. Process according to claim 10, characterized in that the taxol and the compound of general formula II react in 1: 1 molar ratio, based on equivalents, to form a 2'-subsubstituted compound of general formula II, wherein R is a group of general formula III: -c- &lt; ch2 &gt; "- no; (im R3 and R 'is hydrogen. 12. Process according to claim 10, characterized in that 2 equivalents of the compound of general formula II react with 1 taxol equivalent and form a 2-7-di- (substituted) compound of the general formula. I, wherein R and R 'each represent a group of general formula III. Process according to claim 12, characterized in that the compound 2', 7-di- (substituted) is subjected to the cleavage reaction for the 2 'cleavage. -substituent and the formation of compound 7- substituted with general formula I, wherein R represents hydrogen and R 'represents a group of general formula III. Process according to claim 10, characterized in that the reaction is carried out in the presence of an agent of 109072. The process according to claim 13. characterized in that the cleavage reaction consists in subjecting the compound 2 to 7 (di) (substituted) at a pH of 7. .. 7.4 16. Process for preparing a compound of general formula I, wherein R and R 'represent each R or the remainder of an amino acid, provided that at least one of R and R? not be methanesulfonic acid, characterized in that the taxol reacts with an N-protected amino acid and then deprotects the amino acid radical. 17. Process according to claim 16. characterized in that the N-protected taxol and amino acid react in 1: 1 equivalent ratio, forming the substituted 2 'taxol derivative. 18. Process according to claim 16, characterized in that the N-protected taxol and amino acid react in an equivalent ratio of 1: 2, forming the 2V7-disubstituted taxol derivative. 19. A process according to claim 18, characterized in that the compound of general formula I is subjected to a cleavage for cleavage of the 2'-amino acid substituent and formation of the taxol derivative 7- (amino acid). The method according to claim 16. characterized in that the deprotection step is performed by a gentle acid treatment. 21. A process for preparing a taxol derivative of general formula I, wherein R is the remainder of an amino acid, selected from the group consisting of: alanine, leucine, isoleucine, valine, phenylalanine, proline, lysine and arginine or a group of general formula III, wherein n is an entire shoulder between 1 and 3 and R 1 and R 5 are each hydrogen, an alkyl radical having 1 to 3 carbon atoms or together with the nitrogen atom to which they are bonded form a heterocyclic ring saturated with 4 or 5 carbon atoms, characterized in that: (a) taxol reacts with a compound protecting the hydroxyl group to form 2 '- (protected) - taxol: (b) 2' - (protected) - ta-xol reacts with alanine, leucine, isoleucine, valine, phenylalanine, proline, lysine or arginine or with the compound of general formula II, to form 2 '- (protected), 7- (protected) -taxol, followed by (c ) deprotection of position 2. 22. Process according to claim 21, characterized in that the step of deprotection consists of the reaction of 2, - (protein-jat), 7- (protected) -taxol with a mixture of zinc and acetic acid.

Chairman of the invention committee: chem. Novac Maria Examiner: Engineer Marin Elena

Group 12

PreJ lei ^ £) Q

State Office for Inventions and Trademarks

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