DK144599B - METHOD OF ANALOGUE FOR THE PREPARATION OF WINCALCALOID DERIVATIVES AND SALTS THEREOF - Google Patents

Description

in 146599

The present invention relates to an analogous process for the preparation of novel vinca alkaloid derivatives of the general formula IV of claim 1 and pharmaceutically acceptable salts thereof, and the process according to the invention is characterized by the characterizing part of claim 1.

Several naturally occurring alkaloids recoverable from Vinca rosea have been found to be effective in the treatment of experimentally induced malignant conditions in animals. Among these are leurosine (U.S. Patent No. 3,370,057), vincaleukoblastin (vinblastine), hereinafter referred to as VLB (U.S. Pat.

3,097,137), leurosidine (vinrosidine) and leurocristine (VCR or vineristin), both in United States Patent Nos. 3,205,220, 4f-deoxy-VLB - "A" and "B", Tetrahedron Letters, 783 (1968) ( desacetyl-leurosine hydrazide is also mentioned herein); 4-desacetoxy-vinblastine (U.S. Patent No. 3,954,773); 4-desacetoxy-3 * -hydroxyvinblastine (U.S. Pat.

3,944,554); leuroclobin (U.S. Patent No. 3,890,325), leuroformin (N-formylleurosine, see Belgian Pat.

811 110) and vincadiolin (U.S. Patent No. 3,887,565).

Two of these alkaloids, VLB and leurocristine, are now marketed as drugs for the treatment of malignant conditions in humans, particularly the leukemias and related diseases.

The dimeric alkaloids prepared from Vinca rosea can be depicted by the general formula I: 144599 2 i; ii η; ο ·] θ / \ F "f 1 | ΑΛλ /:..^ vr 'R2: C-O-CH3

II

In formula I wherein R is acetoxy, R is methyl, R is hydroxyl, R is ethyl and R is H, VLB is depicted; there are 1 to 3 4 as R is acetoxy, R is formyl, R is hydroxyl, R is 5 L ethyl and R is H, vincristine is depicted; if R is acetoxy, R is methyl, R is ethyl, R is hydroxyl, and R5 is H, leurosidine is depicted; if R 1 is acetoxy, R 2 is methyl, R 3 and R 5 are H and R 4 is ethyl, 4 * - 12 is deoxy VLB "A" depicted; if R, R and R are the same as in 41-deoxy-VLB- "A", but R is ethyl and 4 XO R is hydrogen, 4'-deoxy-VLB- "B" is depicted; and then, as R is acetoxy, R is methyl, R is ethyl, and R and R 3 taken together form a ¢ (epoxide ring, leurosine is depicted.

Of the above alkaloids, vincristine is the most useful and at the same time the least recoverable from Vinca. Jovanovics et al., U.S. Patent No. 3,899,493, have recently developed an oxidative method for converting the relatively more abundant VLB to vincristine vced chromic acid oxidation at low temperatures (-60 ° C). In the vinca alkaloid fraction containing dimeric indole dihydroindoles there are other relatively abundant alkaloids such as leurosine and it would be desirable to convert these directly or indirectly to vineristin or to a drug of similar tumor-degrading effect.

It is well known that leurosine can be converted to 4'-des-oxy-VLB - BM (along with alternating amounts of 4'-deoxy-VLB-An) by treatment with Raney nickel under reflux in absolute ethanol. , see Neuss, Gorman, Cone, and Huckstep, Tetrahedron Letters 783-787 (1968) While leurosin showed tumor-degrading effect on experimentally induced tumors in animals, the clinical response was limited. 4, -desoxy-VLB - ,, A and 4 * -desoxy-15 VLB- "B" was reported as having no reproducible effect against experimentally induced tumors in mice.

It has now been found that, by the process of the invention, compounds of formula IV obtained from 4'-deoxy-VLB- "A" and "B" or their 1-formyl derivatives differ from 4'-deoxy-VLB- " A "and" B "have tumor-degrading effect and can thereby be used as drugs as described below.

A compound of the general formula I wherein is ethyl, R 2 is acetoxy, R is CHO, and R and R are hydrogen, are designated 41-deoxyvinistristine; a compound wherein R 1 is hydroxy while the other groups are the same is designated as 4, -desoxy-4-desacetylvincristine. Since vineristin-like alkaloid is not known which has the opposite configuration of hydrogen and ethyl at the 4'-carbon z Λ 30 atom, such compounds wherein R is ethyl and R is hydrogen are referred to with reference to the compound leurosidine. which have the same configuration at the 4 'carbon atom as 4, -desoxy-VLB -,' B ', and they will be referred to as derivatives of 1-formylleurosidine; eg. 4-35 deoxy-1-formylleurosidine (or 41-deoxyepivincristine) 4 164539 and 4, -desoxy-4-desacetyl-1-formylleurosidine, wherein are acetoxy and hydroxy, respectively. In each of the names given above, it should be understood that the 1-methyl group in leurosidine has been replaced by a formyl group, 5 and that the term "1-desmethyl" has been omitted to simplify the nomenclature.

Nontoxic acids useful for forming pharmaceutically acceptable acid addition salts of the compounds of the invention include salts derived from inorganic acids such as: hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrogen bromide acid, hydrogen iodide acid, nitric acid, phosphoric acid. acidification and the like, as well as salts of non-toxic organic acids comprising aliphatic mono- and dicarboxylic acids, phenyl-substituted alkane carboxylic acids, hydroxy-alkane carboxylic acids, alkanedicarboxylic acids, aromatic acids, aliphatic and aromatic sulfonic acids, etc. pyrosulfate, hydrogen sulfate, sulfite, hydrogen sulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate , propionate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, benzoate, 25 chl orbenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, terephthalate, benzenesulfonate, toluene sulfonate, chlorobenzenesulfonate, xylene sulfonate, phenyl acetate, phenyl propionate, phenylbutyrate, citrate, lacrate, lac Sulfonate, propane sulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate and similar salts.

4, -desoxy-VLB- "A" and "B" or their 1-formyl derivatives are hydrolyzed to the corresponding 4-desacetyl derivatives under acidic or alkaline reaction conditions.

5 144599

The preferred method for carrying out the above hydrolysis reaction uses sodium carbonate in methanol at reflux temperature. Hydrazine hydrate can be used. Other bases which may be used include potassium. 5 t-Butoxide, sodium or potassium methoxide or ethoxide, pyridine, triethylamine (or other tertiary amines), urea and the like in polar organic solvents such as the lower alkanols. Diluted sodium and potassium hydroxide may also be used, e.g. in methanol, but precautions must be taken not to work with base concentrations or reaction temperatures at which other hydrolyzable groups in 4'-deoxyvinocristine or 4'-deoxy-1-formylleurosidine are affected. Bases active only in non-polar solvents may also be used; for example, sodium or lithium hydroxide in benzene, ether, tetrahydrofuran, etc., or the sodium salt of dimethyl sulfoxide in DMSO. Temperatures ranging from room temperature (25 ° C) to the boiling point of the solvent used can be used. On the other hand, the hydrolysis can be carried out under acidic reaction conditions, e.g. in absolute methanol saturated with anhydrous hydrogen chloride at 0 ° C.

The process according to the invention is further illustrated by the following examples.

EXAMPLE 1

Preparation of 4t-deoxy-4-desacetyl-1-formylleurosidine

Ca. 744 mg of 4, -desoxy-1-formylleurosidine are mixed with 10 ml of anhydrous methanol and the mixture is heated to reflux temperature at which temperature the compound 30 is dissolved to clear solution. 200 mg of solid sodium carbonate is added and the reaction mixture is kept under stirring for approx. 7.2 hours at which time TLC of the untreated reaction components shows that virtually all 4, -desoxy-1-formylleurosidine has disappeared. The solvent is removed by evaporation and the residue containing 4 * -desoxy-4-desacetyl-1-formylleurosidine formed by the reaction described above is partitioned between water and methylene chloride. The organic layer is separated and dried, and the solvent is removed by evaporation, which leads to 506 mg of a white solid consisting largely of pure 4'-deoxy-4-desacetyl-1-formylleurosidine.

The compound exhibits the following physical characteristics: Mass spectrum: m / e 766 (M +), 764, 735, 254, 252, 205, 138.

IR spectrum: λ) (CHCl 3), 3450, 1734, 1680, 1596, 1495, 1456, 1434 cm -1.

100 MHz NMR spectrum: (CDCl3) comprises N-formyl at 0.8: 80, methyl single band at 3.89 (C --OCH₂) and 3.66 (C18--CO₂CH₂), 15 broad multiple bands at 3.82 (C ^-CO₂CH₂), and no N-CH₂ about 2.75 (or OCOCH₂, about 2.06).

The corresponding sulfate salt is formed using acetone as the solvent and 0.26 ml of 2% sulfuric acid in ethanol. Other solvents may be used, and it is preferred to use a solvent in which the base is readily soluble but in which the sulfate salt is substantially insoluble.

4'-deoxy-4-desacetylvincristine and its sulfate salt are prepared in a completely analogous manner from 4'-deoxyvinocristine.

EXAMPLE 2

Preparation of 4'-deoxy-4-desacetyl-leurosidine

A reaction mixture was prepared containing 1.48 g of 4'-deoxy-VLB "B", 1 g of sodium carbonate and 100 ml of methanol and heated to reflux under nitrogen atmosphere. Thin-layer chromatography on a representative sample taken after two hours indicated that the hydrolysis reaction to remove the 4-acetyl group was about half complete, the reaction mixture, after standing overnight at room temperature, was heated again to reflux temperature for 8 1/2 hours. Thin layer chromatography of a representative sample using a 20: 1: 1: 1 ether / diethylamine / toluene / methanol solvent system indicated that the reaction had been completely completed. The solvent was removed from the reaction mixture by evaporation and the resulting evaporation residue was dissolved in a mixture of methylene chloride and water. The methylene chloride phase was separated and dried. Evaporation of the methylene chloride led to an evaporation residue, which by TLC was found to contain a highly polar compound plus the expected 4'-deoxy-4-desacetylluerosidine. The residue, which weighed 1.33 g, was dissolved in benzene. The highly polar material was largely insoluble in benzene and was separated by filtration. The filtrate was evaporated to dryness and the residue, weighing 500 mg, was chromatographed on Woelm silica gel using a 20: 1: 1 ether / diethylamine / toluene solvent system (with increasing methanol content) as eluent. The course of chromatography was followed by thin-layer chromatography and fractions found to contain 4'-deoxy-4-desacetylleurosidine were pooled to give 348 mg of base after evaporation of the solvent. The residue was treated with 1.28 ml of 2% sulfuric acid 30 in methanol (0.36 mol) and the solution thus obtained was filtered to give 315 mg of 4'-deoxy-4-desacetylurosidine sulfate.

4'-deoxy-4-desacetylleurosidine exhibited the following physical characteristics:

Mass Spectrum: m / e 752 (M +), 750, 693, 691, 555, 338, 240, 158.

IR spectrum: (CHCl3) 5455, 1724, 1610, 1497, 1457, 1431 cm -1.

100 MHz NMR spectrum: δ 9.43 (br s, 1, C C -OH), 7.92 δ (brs, 1, indole NH), 7.47-7.65 (m, 1, Ci; 1, -H), 7.06-7.51 (m, 3, O, 2ΐ_ΐ4! -Η, 6.58 (s, 1, C ^-H), 6.10 (s, 1, C ^ y-H) ), 5.78-5.87 (m, 2, Cg ^H), 4.10 (m, 1, C4 ~H), 3.83 (s, 3,

Cl6-OCH3), 3.78 (s, * 3, C3 -CO2CH3), 3.70 (s, 1, C2-H), 3.58 (s, 3, C18, -CO2CH3), 2.75 ( s, 3, N-CH 2), 0.76-1.06 (m, 6, C 21.21

The compounds of the present invention which can be imaged by the above Formula IV are more powerful anti-tumor agents. In detecting the action of these agents against mouse transplanted tumors, a method was used which involved intraperitoneal administration of the agent at a predetermined dosage level for 7 to 10 days after the inoculation of the tumor, or alternatively on the first, fifth and ninth day after inoculation.

Table 1 shows the results of a number of experiments in which transplanted tumors in mice were successfully treated with a compound of the invention.

In the table, column 1 shows the name of the compound, column 2 the transplanted tumor, column 3 the dosage level, or dosage level range, and the number of days in which the dosage was administered; column 4 the administration method, and column 5 the percent inhibition of tumor growth or the percentage extension of survival time, e.g. for B16. (ROS is an abbreviation for Ridgeway's osteogenic sarcoma; GLS for Gardner Lympho-sarcoma; P1534 (J) L1210 is leukemia; CA755 is an adenocarcinoma; and B16 is a melanoma).

9 144599

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By using the present novel compounds as anti-tumor agents, they can be administered either parenterally or orally. In oral dosing, a suitable amount of a pharmaceutically acceptable salt 5 is mixed with a base formed with a non-toxic acid such as the starch sulfate salt or other excipient and the mixture is loaded onto telescopic gelatin capsules each containing between 7.5 and 50 mg of active ingredients. Similarly, the anti-neoplastic active salt can be mixed with starch, a binder and a lubricant, and the mixture can be switched to tablets each containing from 7.5 to 50 mg of salt. The tablets may be partitioned if smaller or partial dosages are to be used. However, parenteral administration is preferred. For this purpose, isotonic solutions containing 1-10 mg / ml of salt such as the sulfate salt are used. The compounds are administered to an extent of from 0.1 to 1 mg / kg, preferably from 0.1 to 1 mg / kg, animal body weight once or twice a week or every other day, depending on both the activity of the agent and its toxicity. An alternative method for determining the therapeutic dosage is based on the body surface area, and consists of doses of between 0.1 and 10 mg / m of animal algae surface every 7 or 14 days.

In the clinical application of a compound of the invention, the treating physician will initially administer the compound in the same manner and in the same medium, and probably against the same tumor types as are indicated for vineristin or VLB. The dosage levels used would reflect the difference in dosage level found in the treatment of experimentally induced tumors in mice, the dosage levels of the compounds of the invention being significantly lower than those used with vineristin and VLB. As with other anti-tumor agents, clinical trials will pay particular attention to the oncolytic (tumor

Claims (4)

144599 removing effect of the compounds of the invention against the ten "indicator" tumors listed on page 266 of "The Design of Clinical Trials in Cancer Therapy", edited by Staquet (Futura Publishing Company, 1973). I. Analogous Process for the Preparation of Vinca Alkaloid Derivatives of General Formula IV Pftf- ·, · / "ΡΫψ" "14" "fCO-CHs H] 0 17! ® / * \ 1 Ί! / \ / * / Hs p14 | pr-4 ST ..... CH * * ^ · 0Η in I r1: CO-CHs II 0 wherein R is CH, or CHO; and wherein one of the groups 3 4 ^ and R is H and the other is and pharmaceutically acceptable salts thereof, characterized in that, in a reaction mixture, a vineal alkaloid derivative of the general formula V 144599 is hydrolyzed. 10 ° C ly Λ '· Y VY Y "VyY" · H [O! e / KI prl 1 1A 0.8 CfteO-V15 A, Μ, 1-OC-Ctts RA K CO-CHs II 0 2 3 4 wherein R, R and R have the meaning given above, and recovering the free 4-desacetyl base or a pharmaceutically acceptable one t salt thereof. Process according to claim 1, characterized by hydrolyzing with sodium carbonate. Process according to claim 1 or 2, characterized in that the hydrolysis is carried out in anhydrous methanol. Process according to claims 1-3 for the preparation of 4'-deoxy-4-desacetyl-1-formyl-leurosidine, characterized in hydrolyzing 4'-deoxy-1-formylleurozidine with sodium carbonate in anhydrous methanol.