HU181745B - Process for producing new vinblastin and leurosidin derivatives - Google Patents

Abstract

A process for the preparation of new cytostatic compounds of general formula (I> <IMAGE> in which either R<4> represents a beta -hydroxyl group and R<3> represents an alpha -ethyl group or R<4> represents a hydrogen atom and R<3> represents a beta -ethyl group; R'' represents a methyl group, a straight chain alkyl group having from 3 to 10 carbon atoms, a branched chain alkyl group having from 3 to 10 carbon atoms and in which the carbon atom attached to the <IMAGE> group has a primary or secondary configuration, or an aralkyl having from 1 to 3 carbon atoms in the alkyl moiety; R<1> represents a methoxy group; and R<2> represents an acetyl group; which comprises reacting a compound of formula (II), <IMAGE> (in which R<1>, R<2>, R<3> and R<4> are as defined above), or an acid addition salt thereof with a large excess of an alcohol of formula R''-OH (III> (wherein R'' is as defined above) in the presence of chromium trioxide at a temperature of from -60 DEG C to -30 DEG C.

Description

The present invention relates to a process for the preparation of new compounds of formula I in formula I

R ^{4 is a} hydroxyl of β-position;

R ^{3 is an} alpha-ethyl group, or

R ^{4 is} hydrogen;

R ^{3 is a} β-ethylene group,

R 'other than ethyl having from 1 to 10 carbon atoms, or

C 3 -C 10 represents a branched chain alkyl group in which the carbon atoms attached to the > N _a -CH ₂ -O group have a configuration other than tertiary or a alkyl group containing 1 to 3 carbon atoms in the alkyl moiety -.

The novel compounds of formula (I) of the present invention are cytostatic.

The compounds of formula (I) may be prepared from the acid addition salt of the corresponding compound of formula (II), or the corresponding compound of formula (II) itself by oxidation in the presence of a compound of formula (III). In formula (II), R ³ and R ⁴ and in formula (III) R 'are as defined for formula (I). The reaction essentially involves a structural modification of the z: N _a -CH ₃ group of _the compound of formula II, which can be accomplished with virtually any bis-indole alkaloid containing _the N- _a- CH ₃ moiety. 30

Starting materials of the formula II are known. Isolation of vinblastine R ³ = α-ethyl and R ⁴ = O-hydroxyl (US 3,097,137 and 20'-deoxy leurosidine (R ³ = β-ethyl and R ⁴ = hydrogen) N. Neuss et al., Tetrahedron Letters, 1968, 783, (synthesis by Potier et al., J. Am. Chem. Soc., 98, 7017, 1976) .The known starting materials themselves have a cytostatic effect, and new derivatives derived from them are also cytostatic. but less toxic than the starting materials.

The acute toxicity of the compounds was investigated in groups of 6-6 mice of male Swiss mice weighing 27-31 g from their own culture. Test substances were prepared by injection of saline and, if necessary, addition of 1 drop of Tween 80. Injection solutions were administered intraperitoneally in non-lethal doses up to 100% lethal dose, and results were calculated by the method of Lichfield and Wilcoxon. The results in comparison with known vinblastine, vindezine and vincristine are shown in the following table.

Thus, the new compounds are 15 to 25 times less than vincristine and vindezine, and vinblastine

They were 8 to 13 times less toxic and, unlike vincristine and vindesine, had no paralysis until LD _S0 was reached.

·>

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Compound LDjo (mg / kg, ip mouse) Side effect with paralysis * N-demethyl-N- (methoxymethyl) vinblastine ~ 100 Φ N-demethyl-N- (propoxy methyl) vinblastine ~ 80 Φ N-demethyl-N- (isobutoxy methyl) vinblastine ~ 60 Φ N-demethyl-N- (heptoxy methyl) vinblastine ~ 60 Φ N-demethyl-N- (benzyloxy methyl) -vinbalsztin ~ 70 Φ vincristine 4.2 + vinblastine 7.6 Φ vindesine 4.0 +

* + is;

♦ 0: None

The cytostatic activity of the new compounds was also examined in tissue culture and in various transplanted tumor strains.

The test substances were dissolved in tissue culture (HeLa cell culture) (from a threshold dose of 1 · 10 ^-3 Mg / ml to an upper dose limit of 100 Mg / ml) and in 24-hour cultures by in vitro microscopic examination of cells arrested in metaphase:

Compound Strong block inducing dose (gg / ml) 5 N-demethyl-N- (methoxymethyl) - -vinbalsztin 0,001 N-demethyl-N- (propoxymethyl) - vinblastine 0,001 10 N-demethyl-N- (isobutoxymethyl) - vinblastine 0,001 N-demethyl-N- (heptoxy-ylmethyl) - vinblastine 0,001 N-demethyl-N- (benzyloxy-methyl) - 15 vinblastine 0.01

Under similar conditions, but using stained tissue cultures20, it was possible to observe subtle differences that can be classified into five steps. namely

Grade 1: The minimal effect dose is characterized by an increase in the ratio of blocked mitoses. Some of these may be distorted, such as 3-group mitosis, or a few chromosomes separated on the pole. The anaphases usually disappear. In the second stage, when a strong metaphase block occurs, there is practically no regular mitosis. The chromosomes are located in a loose cotyledon. The proportion of interphase cells is low. The third level of strength when it comes to chromosomes

Compound Dose in mg / kg ip Avg. Life (days) Managed (control) toxicity handled control N-desmethyl-N-methyl-heptoxy vinblastine 8x0,4 14.3 10.3 139 8x4,0 18.7 10.3 181 were not detected 8 x 8.0 21.3 9.9 217 N dezrnetil-N- (benzyloxy-methyl) - 8x0,4 13.0 10.3 126 vinblastine 8x4,0 18.3 10.3 178 were not detected 5x8,0 19.2 9.9 195 N-demethyl-N- (isobutoxymethyl) - 8x0,4 13.7 10.3 133 vinblastine 8x4,0 20.0 10.3 194 were not detected 8x8,0 20.5 9.9 208 N-demethyl-N- (methoxymethyl) - 8x 1.0 12.7 10.3 122 vinblastine 8x2,0 18.7; 16.8 10.5 178; 155 were not detected 8x4,0 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-demethyl-N- (propoxymethyl) - 8x 1.0 15.0 10.3 145 vinblastine 8 x 2.0 20.8 10.5 198 8 x 4.0 19.0 n, i 171 8x8,0 20/2 9.9 205 8 x 10.0 14.7 10.3 142 toxic*

♦ The animals died tumor-free.

-2181745 are coalesced together in a single dense mass at the center of the cell. This is pycnomatosis, or "Ball-Metaphase." The fourth stage, when the effect is already on the interphase cells, is that relatively few blocked 5 cells are able to enter mitosis. The cells have an expanded cytoplasm (interphase), the cytoplasmic border is irregular, "fringed", and the cell often has an elongated fiborblast-like shape. Finally, in the fifth stage, the cytoplasm is filled with a fine reticular structure, and it is clear that the interphase cell was almost fluxed by treatment. These grades were well distinguished for the current materials.

Based on the above, the most potent substance was found to be the heptoxic derivative. This has already produced a strong metaphase block at a dose of 0.001 Mg / ml. Pinomatosis occurred at doses of 0.1 mg / ml and interphase cell activity occurred at doses of 1 to 10 mg / ml. The isobutoxy derivative was one order of magnitude weaker: regular mitoses still occurred at the lowest dose of 0.001 Mg / ml. Finally, the benzyloxy derivative, which produced a minimal block at the lowest dose, with some distorted mitoses, but no anaphases, was the weakest. Medium block at 0.01 Mg / mL; strong block and pycnomatosis occur only at a dose of 1 mg / ml.

The effect of the new compounds was i.p. Transplantable tumor cola (P388 murine leukemia and NK / Ly ascites lymphoma) is described below.

P388 leukemia is maintained in BDFj hybrid mice, and groups of 6-6 mice are assayed with 10 x tumor cell / animal ip. transplanted. Begin 24 hours after transplantation with the new compounds daily ip. and animals are monitored daily for weight and condition. The effect on the treated animals is expressed as a percentage of the average survival time of the control group in days.

It can be seen from the above table that the derivatives tested all significantly prolong the life span of P388 leukemic mice.

Groups of Swiss-H / Riop uotbred mice from our own culture were transplanted with 5 x 10 ⁶ ascites tumor cells ip. Treatment was initiated 24 hours after transplantation and the compounds were administered a total of five times daily. The mean lifespan of the control group was 15.7 days.

The effect of the compounds is manifested in significant prolongation of life, as shown in the following table.

Compound Dose in mg / kg ip. Living on Day 25 - tumor on the 25th day on Day 30 N-demethyl-N- (heptoxy methyl) vinblastine 5 x 6.0 10/10 7/10 3/10 N-demethyl-N- (isobutoxymethyl) vinblastine 5x4,0 10/10 10/10 7/10

The novel compounds exhibit antitumor activity at doses of 4-8 mg / kg / day on P388 and NK / Ly strains and are equivalent to known diindole alkaloids. However, they are significantly less toxic than known compounds of similar structure.

For human use, the compounds are preferably administered intravenously or by infusion.

The present invention therefore relates to a process for the preparation of novel compounds of formula (I)

R ^{4 is a} 0-hydroxyl group, and

R ^{3 is an} alpha-ethyl group, or

R ⁴ hydrogen atoms &

R ³ 0-spacing is ethyl, R "is other than etiltől C1-10 straight or

C 3 -C 10 represents a branched alkyl group in which _the carbon atom attached to Z ₂ N _a - CH ₂ -O has a non-tertiary configuration or the alkyl moiety contains 1 to 3 carbon atoms, such that a (II) ) general formula vegyü45 kick or acid addition salt thereof - in which R ³ and R ⁴ are as defined above - alcohols of general formula (III): - in formula (III) R "a are as defined above - from 50 to 150-fold molar excess of chromium trioxide, an organic solvent, in the presence of acetic anhydride such as acetic anhydride and an acid at a temperature of from -60 ° C to -30 ° C, preferably from -50 ° C to -60 ° C, and adjusting the pH of the reaction mixture to 8-10 the resulting product of formula (I) is purified if desired and then recovered.

The process according to the invention is based on a compound of formula II or an acid addition salt thereof.

The starting material is dissolved in a suitable organic solvent, preferably acetone, or a chlorinated hydrocarbon, preferably non-alcoholic dichloromethane, chloroform or dichloroethane. The presence of an acid facilitates both dissolution and reaction. It is advisable to work in a slightly acidic environment.

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Preferably, an organic acid such as glacial acetic acid, adipic acid, pelargonic acid and optionally a small amount of inorganic acid is added to the reaction mixture. An inorganic acid may be an inorganic acid forming an acid addition salt with the compound of formula (II) which may be present in traces or in equimolar amounts.

The alcohol of formula (III) used as the reactant is used in high molar excess. Examples of alcohols include straight chain alkanols having from 1 to 10 carbon atoms, the reaction of which with ethanol is described in Hungarian Patent Application No. 178,706. In particular, methanol, propanol, butanol, amyl alcohol, heptanol may be used. Further, C3-C10 branched alkanols in which the —OH group is attached to a carbon atom other than the tertiary configuration may be used. In the case of tertiary alkanols, the reaction does not take place. Preferably, i-butanol, i-propanol, and the like. employed. Finally, arylalkyl alcohols having from 1 to 3 carbon atoms, such as benzyl alcohol, phenethyl alcohol, may be used.

The alcohols of formula (III) are used in high molar excess. The molar ratio of alcohol to compound of formula (II) varies from alcohol to alcohol, preferably from 50 to 150 molar equivalents of alcohol (III).

The compounds of formula (II) and (III) are reacted in the presence of chromium trioxide (CrO ₃ ). The chromium trioxide is conveniently added in dissolved state, preferably in acetic anhydride solution.

The reaction is preferably carried out in a protective gas atmosphere at a temperature of -60 to -30 ° C, preferably -50 to -60 ° C. The progress of the reaction is monitored by any suitable analytical method, preferably by thin layer chromatography. The reaction is usually carried out over a period of from 20 minutes to 1 hour and 40 minutes, depending on the alcohol (III) used.

After the completion of the reaction, the pH of the reaction mixture is adjusted to 8-10 to completely decompose the acetic anhydride. Preferably, the pH is started at -50 ° C and is usually completed at 0 ° C to + 30 ° C. Preferably, the reaction mixture is stirred for 5 to 10 minutes at room temperature.

The crude product obtained is then recovered by extraction and evaporation. The crude product may also contain a small amount of N-desmethyl and N-desmethyl-N-formyl derivatives, from which the target product is conveniently separated by chromatography. Chromatography is conveniently carried out on partially deactivated alumina or on fine silica gel adsorbent, and the isolated product is purified, if desired, by recrystallization.

The following examples are intended to illustrate the process, but are not intended to limit the scope of the examples.

Example 1

N-demethyl-N- (methoxymethyl) vinblastine

Vinblastine sulfate (1.0 g, 1.1 mmol) in 240 ml of absolute alcohol-free dichloromethane, methanol and 25 ml of glacial acetic acid in a mixture of 8.0 ml, and the solution was cooled to -55 ^e C. A solution of 0.5 g (5.0 mmol) of chromium trioxide in 40 ml of acetic anhydride, cooled to -55 ° C, was added dropwise to the vinblastine solution under vigorous stirring under a stream of dry nitrogen for 5 minutes. Meanwhile, the temperature of the reaction mixture must not exceed -50 ° C. The progress of the oxidation reaction was monitored by TLC (adsorbent: DC- _Alufolien Kieselgel 60 F ₂ S ₄ , Art 5554; eluent: diethyl ether-ethanol-benzene-diethylamine 10: 0.5: 0.5: 0.5). The oxidation usually takes about 20-30 minutes. A mixture of concentrated aqueous ammonium hydroxide (190 mL) and ice (200 g) was added to the reaction mixture while cooling was maintained. The temperature rises from -55 ° C to 0 ° C to + 10 ° C. The external cooling is then removed and the solution at pH 8.5-9 is stirred vigorously for 10 minutes. The phases are separated, the aqueous phase is extracted with dichloromethane (3 x 30 mL), the extract is washed first with four 25 mL portions of 1: 1 dilute aqueous ammonium hydroxide, twice with 30 mL of water, and the organic phase is dried over magnesium sulfate. evaporation in vacuo; 0.85 g of crude product is obtained, of which 0.3 g is chromatographed on a column of activity II-III alumina in dichloromethane. 6 ml of dichloromethane eluate fractions were collected and chromatographed on TLC (adsorbent: DC-Alufolien Kieselgel 60 F ₂₅₄ , Art 5554; solvent: dichloromethane: methanol = 5: 0.4, detection: or 254 nm ultraviolet light).

Fractions containing the same alkaloid were combined and evaporated separately. The target compound is a

6-25. fractions.

Yield: 64% of the title compound, which is chromatographically uniform.

205-210 ° C (ethanol). [Α] D = + 23 ° (c = 1, chloroform).

1 H-NMR (CDCl ₃ , 100 MHz): 59.05 (s, 1H, C ₁₆ -OH), 8.05 (s, 1H, N _a , H), 7.35 (s, 1H, C ₁₂ , -H), 7.13-7.26 (m, 3H, C 1 -Cu> -H), 6.68 (s, 1H, C ₉ -H), 6.33 (s, 1Η, C _{1 Z} - H), 5.84 (d, 1H, C ₁₄ -H),

5.37 (s, 1H, C ₁₇ -H), 5.28 (d, 1H, C _{1 S} -H), 4.42 (2H-N _a -CH ₂ -O-), 4.18 (s, 1H, C ₂ -H), 3.97 (s, 3H, C _{I6 -CO2 CH3),} 3.77 (s, (3H, Cn-0CH _3);

3.62 (s, 3H, C ₁₆ -CO ₂ CH ₃ ), 3.26 (s, 3H, -OCH ₃ ),

2.73 (s, 1H, C ₁ -H ₃ ), 2.09 (s, 3H, OCOCH ₃ ), 0.8-0.96 (2t, 6H, C ₁₈ -H ₃ , C ₁₈ -H ₃ ).

MS m / e 840 (M ⁺ 100%), 810, 809, 781, 751, 681, 651, 650, 601, 499, 355, 282, 243, 241, 154, 149.

Example 2

N-demethyl-N- (isobutoxymethyl) vinblastine

0.5 g (0.55 mmol) of vinblastine sulfate in 120 mL

-4181745 abs. dichloromethane, 3.7 ml isobutanol and 12.5 ml glacial acetic acid, and the solution is cooled to -55 ° C. To the solution was added chromium trioxide (0.25 g, 2.5 mmol) in acetic anhydride (40 ml) cooled to -55 ° C, and the reaction was monitored by thin layer chromatography (adsorbent: DC-Plastikfolien Kiesel) as described in Example 1. 60 F ₂ S ₄ , Art 5735, solvent: ether-ethanol-benzene-diethylamine (10: 0.5: 0.5: 0.5). The reaction is complete in 140 minutes. The reaction mixture was worked up as in Example 1 to give 0.65 g of crude product which was purified by column chromatography on Kieselgel 60 (Art 9385) adsorbent in dichloromethane. Development and elution with dichloromethane were performed in 10 ml fractions. The first 330 ml of eluate does not contain alkaloid. The elution was then continued with 3% methanol in dichloromethane and the first 90 ml of eluate subsequently contained the target compound, which was obtained by evaporation of the eluate.

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

Mp 215-218 ° C (acetone ether).

IR (KBr): 3406, (NH, OH), 1730> (CO ₂ CH ₃ ), 1605, 1220 / cm (OAc). 25 1 H-NMR (CDd ₃ , 100 ΜΗζ); δ8.1 (s, 1H, N _a , H), 7.5 (m, 1H, C ₁₂ , -H), 7.05-7.2 ( 3H, m, C _9, C _1V-H), 6.70 (s, 1H, C ₉ -H), 6.36 (s, 1H, C ₁₂ -H), 5.87 (d, 1H, C _i4-H), 5.37 (s, 1H, _C17 -H), 5.30 (d, 1 H, C H _{1 s),} 4.75 4.15 (2H, J _AB 10 Hz, 2 = N _{-CH 2-30} -O-), 4.0 (s, 1H, C ₂ -H), 3,78,3,75 (2s, 6H, CO ₂ CH _3), 3.63 (s, 3 H , Cu-OCH ₃ ), 2.75 (s, 1H, C ₂₁ H), 2.1 (s, 3H, OCOCH ₃ ), 0.95-0.70 (12H, CH ₃ ).

MS m / e: 882 (M ⁺ , 100%), 864, 851, 823, 810, 779, 35 751, 723, 651, 650, 514, 355, 346, 329, 154.

Example 3

N-demethyl-N- (heptoxy methyl) vinblastine

In the second example, except that 3.7 ml of isobutanol is used as reagent,

3.7 ml of 1-heptanol were used; reaction time approx. 90 minutes. Workup of the reaction mixture gave 4 g of crude product which was purified by column chromatography on Kieselgel 60 (Art 9385) to 30 ml of dichloromethane. Development and elution with dichloromethane. 10 ml of eluate fractions were collected, the first of which did not contain 300 ml of alkaloid. The elution was then continued with dichloromethane containing 3% methanol and the first 120 ml thus obtained contained the target product. Yield: 21% of the title compound. 200-205 ° C (acetone-ether). IR (KBR): 3450 (ΟΗ, ΝΗ), 1740 (CO ₂ CH ₃ ), 1610, 1220 / cm 2 (OAc). ^Χ Η-NMR (CDCl ₃ , 100 MHz): δ8.07 (s, 1H, N _a , H),

7.52 (m, 1H, C ₁₂ -H), 7.2-7.02 (m, 3H, C ₉ -C ₉ Cl-H), 6.70 (s, 1H, C 1 -H), δ , 35 (s, 1H, ^w C ₁₂ -H), 5.85 (dd, 1H, C ₁₄ -H), 5.39 (s, 1H, C ₁₇ -H), 5.30 (d, 1H, C _ls H), 4.75 4.15 (2H, J _AB 10 Hz, _the N -CHj-0-), 4.0 (s, 1H, C ₂ -H), 3.80

3.75 (2s, 6H, CO ₂ CH ₃ ), 3.65 (s, 3H, Cn-OCH ₃ ),

2.75 (s, 1H, C ₂₁ -H), 2.10 (s, 1H, OCOCH 3), 0.7-1.0 (3t, 9H, CH ₃ ).

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-demethyl-N- (benzyloxymethyl) vinblastine

In each case, the procedure of Example 2 was repeated except that 4.3 ml of benzyl alcohol was used as the reagent instead of 3.7 ml of isobutanol. After a reaction time of 45 min, 4 ml of crude oil are obtained, which is isolated by preparative thin layer chromatography (adsorbent: Kieselgel PF ₂₅ 4 + 366 »solvent: 100 ml dichloromethane / 8 ml methanol).

Yield: 75 mg (15%) of the title compound. M.p. 215-218 ° C (methylene chloride ether).

IR (KBr): 3400 (OH, NH), 1740 (CO ₂ CH ₃ ), 1610, 1230) cm (OAc).

1 H-NMR (CDCl ₃ , 100 MHz): δ 8.07 (s, 1H, N _a , H),

7.52 (m, 1H, C ₁₂ -H), 7.2-7.08 (m, 3H,

C ₉ H-CH 3 -H, 3H, aromatic), 6.75 (s, 1H, C ₉ -H), 6.20 (s, 1H, C ₁₂ -H), 5.85 (dd, 1H, C ₁₄ -H), 5.41 (s, 1H, C ₁₇ -H), 5.30 (d, 1H, C ₁₅ -H), 4.80, 4.20 (2H, J _ab 10Hz, = N _{-CH 2} -O-), 4.40 (s, 2H, benzilCH _2), 4.00 (s, 1H, C ₂ -H), 3.70, 3.68 (2s, 6H, CO ₂ CH _3), 3.63 (s, 3 H, Cn-OCHA), 2.75 (s, 1H, C21-H), 2.0 to 0.70 (2t, 6H, _CH3 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, 135th

Example 5

N-demethyl-N- (methoxymethyl) -vmblasztin

5g (5.5 mmoles) of vinblastine sulfate are dissolved in 150 ml of water and concentrated aqueous vinblastine base ₄₅ by adding ammonium hydroxide (pH 8) is exposed, and four times with 50 ml of extracted into dichloromethane. The dichloromethane solution was dried over sodium sulfate, filtered and the filtrate evaporated to dryness in vacuo. 4 g of vinblastine base, which was dissolved in 1000 ml of methyl alcohol _5Q dry dichloromethane, 120 ml of acetic acid and 6 ml of methanol and the solution cooled to -55 ° C. To the resulting solution was added a solution of chromium trioxide (2.5 g, 25 mmol) in acetic anhydride (470 mL) cooled to -55 ° C over about 5 minutes. The reaction mixture was stirred at -55 ° C for 45 minutes and then poured into a mixture of concentrated aqueous ammonium hydroxide solution (940 mL) and ice water (940 mL). Make sure that the temperature does not rise above 30 ° C. The phases are separated and the dichloromethane phase is stirred with 1000 ml of 1% aqueous ammonium hydroxide solution. The organic phase is then separated, dried over sodium sulfate, filtered and the filtrate is concentrated to dryness in vacuo. The crude product (3.8 g) obtained in the reaction is purified by column chromatography. The chroma-5181745 tograph column was formed from 230 g of Brockmann activity II-III alumina and benzene. The crude product was dissolved in 20 mL of 8: 2 benzene-chloroform and layered on the column. Elution is carried out with an equal volume of benzene-chloroform (about 8 L) and 100 mL of eluate fractions are collected. Fractions containing the same material were combined and evaporated (adsorbent: silica gel; eluent: benzene-ethanol-diethylamine-diethyl ether 5: 5: 5: 100) to give 2.1 g of the title compound, which was recrystallized from 6 ml of ethanol.

Yield: 1.5 g of the title compound.

The physical constants are the same as in Example 1.

Example 6

N-demethyl-N- (propoxymethyl) vinblastine

The fifth procedure was followed except that methanol was used as reagent

4.5 ml of propanes were used.

Yield: 0.6 g of the title compound.

200-210 ° C (ethanol). [α] D 20 = + 31.2 ° (c = 1, chloroform).

1 H-NMR (CDCl 3, 100 MHz): δ 9.05, (s, 1H,

C ₁₆ -OH), 8.06 (s, 1H, N _a , H), 7.34 (m, 1H, C ₁₂ -H), 7.34-7.12 (m, 3H, C ), 6.68 (s, 1H, C ₉ -H), 6.35 (s, 1H, C ₁₂ -H), 5.84 (dd, 1H, C ₁₄ -H), 5.36 (s, 1 H, C ₁₇ -H), 5.28 (d, 1H, C _{1 S} -H), 4.75 4.15 (2H, J _AB 10 Hz, = N _a -CH ₂ -O-), 4.18 (s, 1H, C ₂ -H), 3.78 (s, 3H, CO ₂ CH ₃ ),

3.75 (s, 3H, Cn-OCHs), 3.62 (s, 3H, CO ₂ CH ₃ ),

2.73 (s, 1H, C ₂₁ -H), 2.09 (s, 3H, OCOCH ₃ ), 0.65-0.94 (m, 9H, CH ₃ ).

Example 7

N-demethyl-N- (methoxymethyl) vinblastine

The procedure of Example 1 was followed except that vinblastine dihydrochloride was used as starting material instead of vinblastine sulfate.

Yield: 28%.

The physical constants are the same as in the first example.

Example 8

N-demethyl-N- (methoxymethyl) vinblastine

The procedure is analogous to that described in Example 1 except that the solvent used is dichloromethane instead of alcohol-free chloroform, dichloroethane or acetone. Yield: 4C-50% of the title compound (yield varies depending on the solvent used). The physical constants are the same as in Example 1.

Example 9

N-desmethyl-N- (methoxymethyl) -20'-deoxy-leurosidine mg (0.067 mmol) 20'-deoxy-leurosidine sulfate - catalyzed by 3 ', 4'-anhydro-vinblastine (Pd / C, methanol) hydrogenation followed by formation of the sulfate salt) 20 ml of abs. dichloromethane, 1.5 ml glacial acetic acid and 0.5 ml methanol. The solution was cooled to -55 ° C and a solution of chromium trioxide (30 mg, 0.3 mmol) in acetic anhydride (3.5 ml) cooled to -55 ° C. The reaction is complete at -55 ° C for 60 minutes. The reaction mixture was then treated with a mixture of concentrated aqueous ammonium hydroxide solution (12 mL) and ice (12 g), and the layers were allowed to separate. After separation of the phases, the aqueous phase is extracted three times with 10 ml of dichloromethane each time, the combined organic phases are washed twice with 10 ml of a 1: 1 dilute aqueous ammonium hydroxide solution and twice with 5 ml of water, and the organic phase is evaporated; 48 mg of crude product are obtained, which is purified by column chromatography (adsorbent: silica gel 0.040-0.063 mm, Merek, Art. 9385; solvent: CH ₂ Cl ₂ then 3? MeOH-CH ₂ Cl ₂ ). 18 mg (32%) of the title compound are obtained.

185-190 ° C (dec.).

IR (KBr): 3400 (NH), 1720 (CO ₂ CH ₃ ), 1610, 1230 / cm (OAc).

1 H-NMR (CDCl 3): δ7.95 (s, 1H, N _B , '- H), 7.54 (m, 1H, C ₁₂ , -H), 7.2-7.1 (m, 3H, ( (CH 2 H), 6.56 (s, 1H, C ₉ -H), 6.11 (s, 1H, C ₁₂ -H), 5.87 (dd, 1H, C ₁₄ -H), 5.46 (s, 1H, _C17 -H), 5.37 (d, 1H, C H _s), 4.72, 4.15 (2H,> N _{-CH 2} -O-), 3.80,

3.75, 3.61 (9H, CO ₂ CH ₃ , Cn-OCHs), 2.11 (s, 1H, OCOCH ₃ ), 1.0-0.6 (6H, CH ₃ ).

[α] D = + 50 ° (c = 1, chloroform).

Claims (9)

A process for the preparation of new compounds of the formula I in the formula I R ^{4 is a} 0-hydroxyl group, and R ^{3 is an} alpha-ethyl group, or R ^{4 is} hydrogen; R ³ 0-ethyl group, R 'other than ethyl having from 1 to 10 carbon atoms, or Means a branched alkyl group having from 3 to 10 carbon atoms, wherein the carbon atom attached to the r: N _a -CH ₂ -O- group has a configuration other than tertiary, or the alkyl part contains from 1 to 3 carbon atoms, characterized in that A compound of formula (II) or an acid addition salt thereof, wherein R ³ and R ⁴ are as defined above, an alcohol of formula (III) wherein R 'is as defined above, 5 to 15 times molar; with excess chromium trioxide, an organic solvent and acetic anhydride, and in the presence of an acid, between -60 ° C and -30 ° C, preferably -50 ° C, and After reaction at a temperature of -60 ° C, the reaction mixture is adjusted to a pH of 8-10, and the product of formula (I) obtained is purified if necessary and recovered. 2. The process of claim 1 wherein the organic solvent is a chlorinated hydrocarbon. 3. The method according to claim 1 embodiment N-demethyl-N- (methoxymethyl) vinblastine front ¹⁰ thereof, characterized in that vinblastine or an acid addition salt thereof is reacted with chromium trioxide in the presence of methanol. 4. The method according to claim 1 embodiment N-demethyl-N- (propoxy-pyridylmethyl) vinblastine front ¹⁵ thereof, characterized in that vinblastine or an acid addition salt thereof is reacted with chromium trioxide in the presence of propanol. 5. The method according to claim 1 embodiment N-demethyl-N- (isobutoxymethyl) vinblastine front ²⁰ thereof, characterized in that vinblastine or an acid addition salt thereof is reacted with chromium trioxide in the presence of isobutanol. 6. The process of claim 1 for the preparation of N-desmethyl-N- (heptoxymethyl) vinblastine, wherein the vinblastine or an acid addition salt thereof is reacted with 1-toluene in the presence of chromium trioxide. 7. A process according to claim 1 for the preparation of N-desmethyl-N-benzyloxymethyl-vinblastine comprising reacting vinblastine or an acid addition salt thereof with benzyl alcohol in the presence of chromium trioxide. 8. The process according to claim 1, wherein N-desmethyl-N- (methoxymethyl) -20 '-deoxy leurosidine is prepared by reacting 20'-deoxy leurosidine or an acid addition salt thereof with methanol in the presence of chromium trioxide. 9. A process for the preparation of a cytostatic pharmaceutical composition, wherein the compounds of formula I according to claim 1, wherein R ³ , R ⁴ and R 'are as defined in claim 1, are pharmaceutically acceptable. in admixture with excipients and / or excipients used in the preparation of such compositions for constitution.