SE442116B - VIEW TO PREPARE CYTOSTATIC COMPOUNDS INCLUDING DERIVATIVES OF VINBLASTIN AND LEUROSIDINE WITHOUT PHARMACEUTICAL PREPARATIONS THEREOF - Google Patents

Description

The compound of the general formula (m or from an acid addition salt thereof, by oxidation in the presence of a compound of the general formula R 2 -OH p (III) In formulas II and III, the general compounds the symbols R1, R2, R3, R "and R" have the same meanings as given above for these symbols in formula I. The reaction essentially comprises a change in the structure of the group:> Na-CH; in the compounds II. is obtained in each bis-indole alkaloid containing a group> Na-caa.

The starting compounds II are known. Vinblastine (a compound of formula II, wherein R 1 represents methoxy, R 2 is acetyl, R 3 is α-ethyl and R 2 is β-hydroxyl) is described in U.S. Pat. , where R1 represents methoxy, R2 acetyl, Ra B-ethyl and R 'hydrogen) as well as iso- -II Éššw fi í ï? c) §j§š 10 15 20 25 30 35 40 3 8104877-9 lation of this compound is described by N. Neuss oa (Tetrahedron Letters 1968, 783) and its synthesis are described by Potier et al .; (J. Am. Chem. Soc. 98, 7017/1976 /). The starting compounds have cytostatic activity but are more toxic than the compounds I prepared therefrom.

The acute toxicity of the compounds of the invention has been tested on male mice (Swiss) weighing 27-31 g.

The tests were performed on groups of 6 mice each. The test compounds were administered intraperitoneally in the form of injectable solutions prepared from physiological saline and optionally a drop of "Tween" 80. The doses were successively increased from a dose which did not cause mortality up to the lethal dose. The results were evaluated using the method developed by Lichfield and Wilcoxon and are summarized in Table 1, where they are compared with the corresponding values for vinblastine, vindesine and vincristine.

Table 1 Compound LD 50 (mg / kg, Paralytic ip mice) side effect N-desmethyl-N- (methoxymethyl) -vinblastine w100 D N-desmethyl-N- (propoxymethyl) -vinblastine w80 N-desmethyl-N- (isobutoxymethyl) ) - vinblastine ~ 60 0 N-desmethyl-N- (heptoxymethyl) - vinblastine 'wo o N-desmethyl-N- (benzyloxymethyl) - vinblastine N70 0 vincristine 4.2 + vinblastine' 7,6 0 vindesine 4.0 + According to According to the results compiled in Table 1, the new compounds are 15-25 times less toxic than vincristine and vindesine and 8-13 times less toxic than vinblastine. In contrast to the case with the administration of vincristine and vindesine, no paralytic side effect has been observed at doses up to LD50 of the compounds of the invention.

The cytostatic activity of the novel compounds of the invention has also been tested in tissue cultures and various transplanted tumor species.

The test compounds were dissolved in the tissue culture (HeLa * cell culture) at concentrations between a threshold dose of 1 x 10 -3 pg / ml up to 100 pg / ml. In 24-hour cultures, the following results were obtained by microscopic evaluation of the metaphase-arresting effect in vivo.

Table 2 Dose that induces strong blocking Compound (pg / ml) N-desmethyl-N- (methoxymethyl) -vin- blastin. 0.001 N-desmethyl-N- (propoxymethyl) -vin-blastin-'0, oo1 N-desmethyl-N- (isobutoxymethyl) -vinblastine 0.001 N-desmethyl-N- (heptoxymethyl) -vinblastine 0.001 N-desmethyl-N- ( benzyloxymethyl) - vinblastine 0.01 Under the same conditions but using sieved cultures, minor differences could also be observed, which can be classified as follows: The first stage (minimum effective dose) is characterized by an increased ratio of blocked mitoses. Some of these are abnormal mitoses, thus, for example, 3-group mitoses or polar chromosomes also occur. In general, anaphases have already disappeared.

In the second stage, a strong metaphase blockage is observed and there is virtually no regular mitosis. Chromosomes are present in the form of a loose glomus. The proportion of interphase cells is low.

In the third stage, the chromosomes have aggregated into a mass in the middle of the cell. This stage is referred to as "Ball-Metaphase" picnomitosis.

In the fourth stage, the interphase cells are already affected. More specifically, the number of blocked cells is reduced, since they cannot participate in mitosis. The cytoplasm of the cells (in the interphase) is increased. Its boundaries are irregular, _- na ... a- ÖOR eyes? in 10 15 20 25 30 35 40 8104877-9 fringed, and the cells often have elongated, fibroplasty-like shape.

In the fifth stage, finally, the cytoplasm is filled with a fine, mesh-like structure and the interphase cell has been unequivocally killed by the treatment.

The above stages could be clearly distinguished in the testing of the compounds of the invention. The following observations were made: The most effective compound was the heptoxide derivative, which induced a strong metaphase blockade already at a dose of 0.001 pg / ml.

Pycnomitosis was observed at a dose of 0.1 ng / ml, while the interphase cells were affected at a dose between 1 and 10 ng / ml. The isobutoxide derivative had an effect of a magnitude lower, because at a dose of 0.001 ng / ml regular mitoses were still observed. The benzyloxide derivative had the weakest effect. At the smallest of the doses applied, this compound induced a minimal blockage with several deformed mitoses but without anaphases. At 0.01 pg / ml the block was moderate and a strong block and pycnomitosis were first induced at a dose of 1 pg / ml.

The effect of the new compounds on intraperitoneally transplantable tumors (P Bßßmusleukemia and NK / Ly ascites lymphoma) is described below. P 388 leukemia tests were performed on hybrid mice (BDF). The tests were performed in groups of 6 mice each and 105 tumor cells per experimental animal were transplanted intraperitoneally. The administration of the test compounds began at the 24th hour after transplantation. The treatment was performed intraperitoneally and the body weight and condition of the experimental animals were determined every day. The effect achieved on the treated experimental animals is stated as a percentage of the average life expectancy in days for the control group.

The results given in the following table show that the life expectancy of mice with leukemia (P 388) is significantly increased by the test compounds. 10 15 20 25 30 35 40 8104877-9 Table 3 Dose Life expectancy Treated / mg / kg (days) control Compound i.p. Treated control% oxicites N-desmethyl-N-hep- 8 x 0.4 14.3 10.3 139 toximethyl-VLB 8 x 4.0 18.7 10.3 181 8 x 8.0 21.3 9 .9 217 N-desmethyl-N- (ben- 8 x 0.4 13.0 10.3 126 syloxymethyl) -vin- B x 4.0 18.3 10.3 178 blastin 5 x 8.0 19.2 9.9 195 N-desmethyl-N- (iso- 8 x 0.4 13.7 10.3 133 butoxymethyl) -vin- 8 x 4.0 20.0 10.3 194 blastin 8 x 8.0 20, 5.9 208 N-desmethyl-N- (meth- 8 x 1.0 12.7 10.3. 122 toximethyl) -vin- B x 2.0 18.7; 16.8 10.5 178; 155 blastin 8 x 4.0 19.0; 17 7 11.1 171; 163 8 x 6.0 19.2 10.5 182 8 x 8.0 18.7; 20.2 10.8 172; 205 N-desmethyl-N- (pro- 8 x 1.0 15.0 10.3 145 poximethyl) -vin- 8 x 2.0 20.8 10.5 198 blastin 8 x 4.0 19.0 11, 1,171 8 x 8.0 20.2 9.9 205 8 x 10.0 14.7 10.3 142 toxic * * The experimental animals died tumor-free.

In groups of 10 each, by inbreeding mice (Swiss-H / Riop Gedeon Richter's own experimental animal breed) 5 x 106 ascites tumor cells were transplanted intraperitoneally. 24 hours after transplantation, treatment was initiated and the compounds were administered daily, a total of five times.

The average life expectancy in the control group was 15.7 days. Similar tests were performed with N-desmethyl-N- (methoxymethyl) -20'-deoxy-leurosidine. In these tests, the doses and the number of treatments were varied. The following results were obtained: Table 4 Dose Life expectancy Treated / mg / kg (days) control Compound i.p. treated control% N-desmethyl-N- (me- 4 X 8 21.8 12.1 180 toximethyl) -20'- 8 x 4 20.8 11.6 180 deoxy-leurosidine 4 x 7 17.1 12.1 142 8 x 1 21.7 12.2 178 8 x 0.5 15.3 12.2 126 The compounds gave a significant increase in life expectancy, as shown in the following table. 10 15 20 25 30 35 40 8134877-9 Table 5 Dose On the 25th day On the 30th day _ mg / kg live tumor-tumor-free Compound i.p. free N-desmethyl-N- (heptoxymethyl) ~ VLB 5 x 6.0 10/10 7/10 3/10 N-desmethyl-N- (iso-butoxymethyl) -VLB 5 x 4.0 10/10 10 / 10 7/10 The tumor inhibitory effect of the new compounds on species P 388 and NK / Ly is significant at a dose of 4-8 mg / kg and day and is equal to the effect of the known diindole alkaloids. At the same time, the compounds according to the invention are much less toxic than the known compounds with analogous structure.

For human treatment, the compounds can best be used intravenously or as infusions.

According to the invention there is also provided a process for the preparation of compounds of the general formula (I) CH 2 HO on "wherein R 3, R", R ", R 1 and R 2 have the meanings given above. According to the invention there are prepared compounds of the general, A g PK The formula I in that a compound of the general formula Rf1 (II) wherein R1, R2, R3 and R "have the meanings given above, or an acid addition salt thereof , is reacted with an alcohol of the general formula w-on '(In) where B "is as defined above, the reaction being carried out in a large excess of the alcohol and in the presence of chromium trioxide, a suitable organic solvent and acetic anhydride as well as also an acid, at a temperature between -60 and -30, preferably between -50 and -GOOC, whereupon the pH of the reaction mixture is adjusted to 8-10 and optionally the product of general formula I is purified and then isolated.

In this process, a compound of general formula II or an acid addition salt thereof is used as a starting material. The starting compound is dissolved in a suitable organic solvent, preferably acetone or a chlorinated hydrocarbon, for example in non-alcoholic dichloromethane, chloroform or dichloroethane. The dissolution and reaction benefit significantly through the presence of an acid. It is therefore convenient to carry out the reaction in weakly acidic medium. Preferably an organic acid, for example glacial acetic acid, adipic acid, geranium acid and optionally a small amount of an inorganic acid is added to the reaction mixture. As inorganic acids, one can use such inorganic acids which form an acid addition salt with the compounds II and this acid can be used in an amount from a trace amount up to approximately equimolar amount.

In Reaction Component III, a large molar excess was used. As alcohol, for example, a straight-chain alcohol with 1-10 carbon atoms can be used. The reaction with ethanol is described in European patent application 0018231.

First and foremost, methanol, propanol, butanol, amyl alcohol and heptanol can be used to advantage. Branched alkanols having 3-10 carbon atoms, in which the hydroxyl group is attached to the primary or secondary carbon atom, can also be used. With tertiary alcohols, the desired reaction can not be carried out. Preferably isobutanol, isopropanol, etc. Finally, arylalkyl alcohols having 1-3 carbon atoms in the alkyl moiety can be mentioned as suitable alcohol reactants. Among these compounds, for example, benzyl alcohol or phenylethyl alcohol can be used.

Alcohols III were used in large molar excess.

The molar ratio of alcohol compound II can be varied depending on the alcohol, but preferably 50-150 molar equivalents of alcohol III are used.

Compounds II and III are reacted in the presence of chromium trioxide. Chromium trioxide is suitably added to the reaction mixture in the form of a solution, preferably in acetic anhydride.

The reaction is conveniently carried out in an inert gas atmosphere at a temperature between -60 and -30, preferably between -50 and -60 ° C. The progress of the reaction is monitored by any analytical method, for example by thin layer chromatography; Depending on which alcohol III is used, the reaction is terminated in between about 20 and about 100 minutes.

When the reaction is complete, the pH of the reaction mixture is adjusted to 8-10 to completely eliminate the acetic anhydride. The pH adjustment is suitably started at a temperature of -50 ° C and generally ends at a temperature between 0 and + 30 ° C. The reaction mixture is preferably stirred at room temperature for 5-10 minutes. 7 The crude product obtained is then isolated by extraction and evaporation. The crude product may contain a small amount of N-desmethyl and N-desmethyl-N-formyl derivatives, respectively, which are preferably separated from the desired product by chromatography. The chromatographic purification is conveniently carried out on a partially deactivated alumina or a granular silica gel used as an adsorbent and the isolated product can, if desired, be further purified by recrystallization.

The following examples further illustrate the invention, but it is of course not limited to the particular details and actions set forth in the examples.

Example 1 N-desmethyl-N- (methoxymethyl) -vinblastine 1.0 g (1.1 mol) of vinblastine sulfate was dissolved in a mixture of 240 ml of non-alcoholic, absolute dichloromethane, 8.0 ml of methanol and 25 ml of glacial acetic acid. The solution was cooled to -SEDC.

A solution of 0.5 g (5.0 mmol) of chromium trioxide in 40 ml of acetic anhydride was cooled to -55 ° C and then added dropwise over 5 minutes to the vinblastine solution under a dry nitrogen atmosphere and vigorous stirring. During the addition, the temperature of the reaction mixture must not rise above -SOOC. The progress of the oxidation reaction was monitored by thin layer chromatography (DC-Alufolien Kieselsgel 60 Fgsr; a mixture of diethyl ether, ethanol, benzene and diethylamine in the ratio 10: 0.5; 0.5; 0.5). In general, the oxidation ends in 20-30 minutes. Then, a mixture of 190 ml of concentrated ammonium hydroxide solution and 200 g of ice was added to the reaction mixture while cooling. The temperature rose from -55 ° C to a temperature between 0 and + 10 ° C. When this temperature was reached, cooling from the outside was stopped and the solution, whose pH was 8.5-9, was stirred vigorously for 10 minutes. The phases were separated and the aqueous phase was extracted with dichloromethane (3 x 30 ml) and the extracts were washed with a mixture of water and ammonium hydroxide 1: 1 (4 x 25 ml) and then with water ( 2 x 30 ml). The organic phase was dried over anhydrous magnesium sulfate, filtered and the filtrate was evaporated in vacuo. 0.35 g of crude product was obtained. This was determined by chromatography on a column filled with 0.3 g of alumina with a degree of activity II-III in a dichloromethane solution. Fractions of 6 ml of dichloromethane each were collected and analyzed by thin layer chromatography (DC-Alufolien Kieselgel 60 F254, art. 5554; solvent: a mixture of dichloromethane and methanol in the ratio 5: 0.4; detection: iodine vapor or UV light with wavelength 254 nm).

All fractions containing one and the same alkaloid were combined and each combination was evaporated separately. The desired end product was included in fractions 6-25.

Yield: 64% of the title compound (by the "title compound" in the present context is meant the compound indicated in the title of the example in question), which was uniform in terms of composition. melting point: zo5-210 ° C (sssssn. [α] D D = + 23 ° (c = 1, chloroform). 1 H-NMR (cnclg, 100 Mim: δ 9.05 (s, m, clvoa), δ, 05 (s, 1H, Na, H), 7.35 (s, 1H, C12, -H), 7.13-7.26 (m, 3H, C9, -C11, -H), 6.68 ( s, 1H, C9-H), 6.33 (s, 1H, C12-H), 5.84 (d, 1H, C14-H), 5.37 (s, 1H, C17-H), 28 (d, 1H, C15-H), 4.42 (ZH,> Na-cH2-o-), 4.18 (s, 1H, cz-H), 3.97 (s, 311, cls-cozcag ), 3.77 (S, 3H, C11-OCH3), 3.62 (S, 3H, C15, -CO2CH3), 3.26 (s, 311., -ocn,), 2.73 (s, m , cu-n), 2.00 (s, an, ococssš, 0.8-0.96 (2t, 6H, C13-H3, C13, -H3).

MS m / e: 840 (M + 100%), 810, 809, 781, 751, 681, 651, 659, 601, 499, 355, 282, 243, 241, 154, 149.

Example 2 N-desmethyl-N- (isobutoxymethyl) -vinblastine 0.5 g (0.55 mol) of vinblastine sulphate was dissolved in a mixture of 120 ml of absolute dichloromethane, 3.7 ml of isobutanol and 12.5 ml of glacial acetic acid and the solution was cooled. to -55 ° C. Then 0.25 g (2.5 mmol) of chromium trioxide in 40 ml of glacial acetic acid cooled to -55 DEG C. was added and the progress of the reaction was monitored by thin layer chromatography. in the manner set forth in Example 1 (DC-Plastikfolien Kiselgel 60 F254; art. 5735; a mixture of ether, ethanol, benzene and diethylamine in the ratio 10: 0.5: 0.5: 0.5). The reaction was completed in 140 minutes, the reaction mixture was further treated as in Example 1 to give 0.65 g of crude product, which was purified by column chromatography on Kieselgel 60 (Art. 9385) as an adsorbent in dichloromethane as solvent. The column was washed and eluted with dichloromethane. Fractions of 10 ml each were collected. The first eluate (330 ml) contained no alkaloid. The elution was continued with dichloromethane containing 3% methanol. The first eluate (90 ml) desired the title compound, which was isolated by evaporation- it contained ning.

Yield: 169 mg (35%) of the title compound.

Melting point: 215-218 ° C (acetone and ether).

IR spectrum (KBr): 3400 (NH, OH), 1730 (CO 2 CH 2), 1605, 1220 (cm / OAc). 1 H-NMR (δ, 100 MHzn δ 8.1 (s, 1: a, Naga), 7.5 (m, 1: a, C 12, -H), 7.05-7.2 (m, 3H, C8, -C11, -H), 6.70 (s, 1H, C9-H), 6.36 (s, 1H, C12-H), 5.87 (d, 1H, Cl, -H), , 37 (s, 1H, C17-H), 5.30 (d, 1H, C15-H), 4.75, 4.15 (ZH, JAB 10 Hz),: Na-cnz-oL 4.0 ( s, m, cz-H), 3.78, 3.75 (za, en, cozcznš, 3.63 (s, 3H, C11-OCH3), 2.75 (s, 1H, C21H), 2.1 (s, 3H, OCOCH 2), 0.95-0.70 (12H, CH 3 groups).

MS m / e: 882 (M +, 100%), 864, 851, B23, B10, 779, 751, 723, 651, 650, 514, 355, 346, 329, 154.

Example 3 N-desmethyl-N- (heptoxymethyl) -vinblastine The procedure described in Example 2 was repeated with the modification that 3.7 ml of isobutanol were exchanged for 3.7 ml of 1-heptanol. The reaction time was about 90 minutes. 4 g of crude product were obtained from the reaction mixture. The purification was performed in a chromatographic column containing Kiesel gel 60 (art. 9385) in 30 ml of dichloromethane. The column was washed and eluted with dichloromethane. Fractions of 10 ml each were collected. The first eluate (300 ml) did not contain any alkaloid. The elution was continued with dichloromethane containing 3% methanol. The first eluate (120 ml) inside, fh ~ f *. fwfïï fi 'Y w 10 15 20 25 30 35 13 8104877-9 held the desired title compound.

The yield was 21%.

Melting point 200-205 ° C (acetone / ether).

IR spectrum (KBr): 3450 (OH, NH), 1740 (CO 2 CH 3), 1610, 1220 (cm / OAc).

1 H-NMR (CDCl 3, 100 MH 2): δ 8.07 (δ, 1H, Na, H), 7.52 (m, 1H, λ 2, -H), 7.2-7.02 (m, 3H, Cs, -C11, -H), 6.70 (s, 1H, C9-H), 6.35 (s, 1H, C12 -H), 5.85 (dd, 1H, Cl, -H), , 39 (s, 1H, C17-H), 5.30 (d, 1H, C15-H), 4.75, 4.15 (2H, JAB 10 Hz,:; Na-CH2-O-), 4 , 0 (s, 1H, C2-H), 3.80, 3.75 (ZS, 6H, COZCH3), 3.65 (s, 3H, C11-OCHQ), 2.75 (s, 1H, C21- H), 2.10 (s, 1H, OCOCH 3), 0.7-1.0 (3t, 9H, CH 3 - group).

MS m / e: 924 (M < + >, 100%), 906, 893, 865, 822, 810, 765, 751, 737, 651, 650, 649, 469, 455, 355, 282, 154, 135, 122: Example 4 N-Desmethyl-4- (benzyloxymethyl) -vinblastine The procedure described in Example 2 was repeated with the change that 3.7 ml of isobutanol were exchanged for 4.3 ml of benzyl alcohol. After 45 minutes, 4 ml of an oily product were obtained, nails were isolated by thin layer chromatography (Kieselgel PF2® + a @ 6; a mixture of 100 ml of dichloromethane and 80 ml of methanol).

Yield: 75 mg (15%) of the title compound. mp 215-21a ° C (methylene chloride / ether).

IR spectrum (KBr) = 3400 (on, NH), 1740 (co, cH @), 1610, 1230 (cm / OAC). 1 H-NMR (cnclg, 100 μm; δ 8.07 (s, 1H, Nam), 7.52 (m, 1H, C 12, -H), 7.2-7.08 (m, 3H, C 9 C11, -H, 3H, aromatic), 6.75 (s, 1H, C8-H), 6.20 (s, 1H, C12-H), 5.85 (da, 1H, C1-H), , 41 (S, 1H, C17-H), 5.30 (d, 1H, C15-H), 4.80, 4.20 (2H, JAB 10 Hz,:> Na-CH2-O-), 4 , 40 (s, 2H, benzyl-CH2), 4.00 (s, 1H, C2-H), 3.70, 3.68 (2s, GH, CO2CH3), 3.63 (s, 3H, C11- OCH 3), 2.75 (S, 1H, C 21 H), 1.0-0.70 (2t, 6H, CH 3 groups).

MS m / e: 916 (M +, 100%), 885, 857, 822, 810, 796, 779, 757, 751, 737, 651, 650, 649, 514, 355, 346, 329, 282, 154, 135 .

Example 5 N-Desmethyl-N- (methoxymethyl) -vinblastine g (5.5 mmol) of vinblastine sulphate was dissolved in 150 ml of water. The vinblastine base was liberated by the addition of a concentrated aqueous solution of ammonium hydroxide (pH 8) and treated with four fractions of 50 ml of dichloromethane each. The dichloromethane solution was dried over sodium sulfate, filtered and the filtrate was evaporated to dryness in vacuo. 4 g of vinblastine base were obtained. This was dissolved in a mixture of 1000 ml of non-alcoholic dichloromethane, 120 ml of acetic acid and 6 ml of methanol. The solution was cooled to -5590. To the solution was added over 5 minutes a solution of 2.5 g (25 mmol) of chromium trioxide in 470 ml of acetic anhydride, which solution was also cooled to -55 ° C. The reaction mixture was stirred at -55 ° C for 45 minutes, then added to a mixture of 940 ml of a concentrated aqueous solution of ammonium hydroxide and 940 ml of ice water. The temperature must not exceed 300 ° C. The phases were separated and the dichloromethane phase was mixed with 1000 ml of a 1% aqueous solution of ammonium hydroxide. The organic phase was separated, dried over anhydrous sodium sulfate, filtered and the filtrate was evaporated to dryness in vacuo.

3.8 g of crude product were obtained. Purification was performed in a chromatographic column prepared from 230 g of alumina with a degree of activity of II-III (Brockmann) and benzene. The crude product was dissolved in 20 ml of a mixture of benzene and chloroform 8: 2 and then layered on the column. Elution was performed with about 8 L of a mixture containing benzene and chloroform in the above proportions.

Fractions of 100 ml each were collected. These were analyzed by thin layer chromatography (silica gel; a mixture of benzene, ethanol, diethylamine and diethyl ether 5: 5: 5: 100). Fractions containing the same substance were combined and evaporated. 2.1 g of the title compound were obtained. Yield (after recrystallization from 6 ml of ethanol) 1.5 g. The product had the same physical properties as the product of Example 1.

Example 6 N-Desmethyl-N- (propoxymethyl) -vinblastine A In the manner set forth in Example 5 but using the reaction component, 0.6 g of the title compound was obtained. ', §? Ml propanol instead of methanol as .w-.T “f 10 15 20 25 30 15 8104877-9 Melting point 200-210 ° C (ethanol).

[Α] D = + 31, z ° (c = 1, chiropractic). 1 H NMR (cncla, 100 mm: δ 9.05 (s, m, ale-one), δ, n6 (s, 1H, Na, H), 7.34 (m, 1H, C12, -H), 7.34 - 7.12 (m, 3H, C9, -C11, -H), 6.68 (s, 1H, C; -H), 6.35 (s, 1H, C12-H), 84 (dd, 1H, Clr-H), 5.36 (s, 1H, C17-H), 5.28 (d, 1H, C15-H), 4.75, 4.1s_ (211, JAB 10 Hz Na-cH2-o->, 4.18 (s, 121, cz-en, 3.78 (s, 3H, COZCH3), 3.75 (s, 3H, C11-OCH3), 3.62 ( s, 33, COZCHB), 2.73 (s, 1H, C21-H), 2.09 (s, 3H, COCH5S6, 0.65-0.94 (m, 9H, CH3 groups).

Example 7 N-desmethyl-N- (methoxymethyl) -vinblastine In the manner set forth in Example 1 but starting from vinblastine dihydrochloride instead of vinblastine sulfate, the title compound was prepared. Yield 28%. The product had the same physical properties as the product of Example 1.

Example gel 8 N-desmethyl-N- (methoxymethyl) -vinblastine In the manner given in Example E but using non-alcoholic chloroform, dichloroethane or acetone as solvent instead of dichloromethane, the title compound was prepared. Yield 40-50% depending on the solvent used. The product had the same physical properties as the product of Example 1.

Example 9 N-desmethyl-N- (methoxymethyl) ~ 20'-deoxyuro-ile oil 60 60 mg (0.067 mmol) of 20'-deoxileurosidine sulphate, prepared by catalytic hydrogenation (Pd / C, methanol) of 3 ', 4'-anhydrovinblastine and subsequent conversion to the corresponding sulfate, was dissolved in a mixture of 20 ml of absolute dichloromethane, 1.5 ml of glacial acetic acid and 0.5 ml of methanol. The solution was cooled to -55 ° C and added with a solution of 30 mg (0.3 mmol) of chromium trioxide in 3.5 ml of acetic anhydride, which solution had also been cooled to -SSOC.

The reaction was completed in 60 minutes. To the reaction mixture was added a mixture of 12 mš og cogšëššíerated water-çïqiíåásgufs and the phases of solution of ammonium hydroxy-phase were shaken with a separate separator. The aqueous chloromethane (3 x 10 ml) is 10104877-9 W 10 15 20 25 30 35 40. The combined organic phases were washed with a mixture of ammonium hydroxide and water 1: 1 (2 x 10 ml) and then with water (2 x 5 ml). Drying and evaporation of the organic phase gave a crude product. Purification was performed in a chromatographic column (silica gel 0.040-0.063 mm Merck, art. 9385; solvent CH 2 Cl 2 followed by a mixture of 3% methanol and CH 2 Cl 2). 18 mg (32%) of the title compound were obtained. melting point 185-19 ° C (sonar eel). [α] D = + 50 ° (c = 1, chloroform).

IR spectrum (KBr): 3400 (NH), 1720 (CO 2 CH 2), 1610, 1230 (cm / OAc). 1 H-NMR (cDc13) = δ 7.95 (S, 1H, Na, -H), 7.54 (m, 1H, C12, -H), 7.2-1.1 (m, sn, ce, -ell, -H), 6.56 (S, 1H, cs-H), 6.11 ia, 1H, C12-H), 5.87 (dd, 1H, Cl, -H), 5.46 ( s, 1H, C17-H), 5.37 (d, 1H, C15-H), 4.72, 4.15 (ZH, I> Na-CH; -O-), 3.80, 3.75 , 3.61 (9H, CO 2 CH 3), 2.11 (S, 1H, OCOCH 3), 1.0-0.6 (GH, CH 3 groups).

Through the above chromatographic purification, 46 mg (22%) of N-desmethyl-N-formyl-20'-deoxyleurosidine was isolated as a by-product.

Melt point 205-210 ° C (decomposition). [α] D = + 54 ° (c = 1, chloroform).

Claims (4)

A process for the preparation of a compound of the general formula wherein R 1 is B-hydroxyl and R 3 is u-ethyl; or R 2 is hydrogen and R a is B-ethyl; R "is a straight chain alkyl group having 1-10 carbon atoms, with the exception of ethyl, or a branched alkyl" group having 3-10 carbon atoms, in which groups the carbon atom attached to the group:> Na-CH2-O- has primary or secondary configuration, or an alkyl group having 1-3 carbon atoms in the alkyl moiety; R 1 is methoxy and R 2 is acetyl, that a compound of the general formula POÜR QUALITY is characterized by a compound in which 8104877-9 R1, R2, Ra and R "have the meanings given above, or an acid addition salt thereof, is reacted with an alcohol of the general formula R" -oH (mi) where R "has the meaning given above, which reaction is carried out in the presence of said alcohol in large excess and in the presence of chromium trioxide, a suitable organic solvent, acetic anhydride and an acid at a temperature between -60 and -30, preferably between -50 and -GOQC, whereupon the pH of the reaction mixture is adjusted to 8-10 and the product obtained is isolated, possibly after it has been purified. 2. A process according to claim 1, characterized in that 50-150 molar equivalents of an alcohol of general formula III are used, based on the amount of compound of general formula II. 3. A method according to claim 2, that a chlorinated hydrocarbon is used as the known solvent for organic solvent .. 10 15 20 25 30 35 19 8104877-9 Pharmaceutical preparation with cytostatic activity characterized in that it contains as active ingredient a pharmaceutically effective amount of a compound of general formula (I) wherein R 4 is β-hydroxyl and R 3 is OC-ethyl; or R 4 is hydrogen and R 3 is β-ethyl; R "is a straight chain alkyl group having 1-10 carbon atoms, with the exception of ethyl, or a branched alkyl group having 3-10 carbon atoms, in which groups the carbon atom attached to the group '> Na-CH 2 -O- has primary or secondary configuration, or an alkyl group having 1 to 3 carbon atoms in the alkyl moiety; R 1 is methoxy and R 2 is acetyl, and at least one pharmaceutically inert substance as carrier or diluent. -, ¿, -..., f- -fi x! ,,, fl umnüwuvøwum = wr, _ ,,, _ .gm 3054 ÅIIPPW ÜÜÜ !! u. -I f .. "" "" "" ~ "_ e.