DK148655B - ANALOGY PROCEDURE FOR PREPARING DERIVATIVES OF 4-DESACETYL-VINCALEUCOBLASTIN-C-3-CARBOX HYDRAZIDE - Google Patents

Description

148555 i

The present invention relates to an analogous process for the preparation of novel derivatives of 4-des-acetyl-vincaleucoblastin-C-3-carboxhydrazide of the general formula I set forth in claim 1, and the process of the invention is characterized by the method of claim 1. l's characteristic part stated.

Several naturally occurring alkaloids recoverable from Vinca rosea have been shown to be effective in the treatment of experimentally induced diseases in 10 animals. Among these alkaloids may be mentioned leurosine (described in U.S. Patent No. 3,370,057), vincaleucoblastin (vinblastine), hereinafter referred to as VLB (described in U.S. Patent No. 3,097,137), leuroformin (described in Belgian Pat. 811 110), leurozidine (vinrosidine) and leurocristine, hereinafter referred to as vineristin (both disclosed in U.S. Patent No. 3,205,220), deoxy-VLB "A" and "B" (described in Tetrahedron Letters, 783 (1958), 4-desacetoxyvinblastine (disclosed in U.S. Patent No. 3,954,773), 4-desacetoxy-3'-hydroxyvinblastine (disclosed in U.S. Pat.

3,944,554), leurocombin (disclosed in U.S. Patent No. 3,890,325) and vincadiolin (disclosed in U.S. Patent No. 3,887,555). Two of these alkaloids, VLB and vineristin, are now marketed as drugs for the treatment of certain diseases, especially leukemia diseases and similar disorders in humans. These two marketed alkaloids are usually administered by intravenous route.

The chemical modification of the Vinca alkaloids has been relatively limited. First, the 30 molecular structures involved are exceedingly complicated, which is why it is difficult to conduct chemical reactions that change a specific functional group in the molecule without affecting other groups. Second, dimeric alkaloids which did not possess the desired chemotherapeutic properties have been prepared from Vinca rosea, and a determination of the structure of these alkaloids has led to the conclusion that these "inactive" compounds are closely related to the active alkaloids. often deviating only from the stereochemistry in a single location. Thus, the anti-neoplastic activity appears to be limited to very specific basic structures, and the possibilities of obtaining more active drugs by modifying these structures appear to be similarly small.

10 Successful attempts to alter physiologically active alkaloids include the preparation of 6,7-dihydro-VLB (disclosed in U.S. Patent No. 3,352,868) and the replacement of the acetyl group by the VLB ring system # 4 with higher alkanoyl groups or with non-related acyl groups (see United States Patent 3,392,173).

Several of these C-4 derivatives are capable of extending the life of mice inoculated with P1534 leukemia.

C-3-carboxamide and C-3-carboxhydrazide derivatives of VLB, vincristine, vincadiolin 20 and other vinca alkaloids have also been prepared, and these derivatives have been found to be effective in the treatment of tumors (see Belgian Patent No. 813,168 and Danish publication no. 141 511). These compounds are extremely interesting, as e.g. The 3-carboxamides of VLB 25 are more effective against Ridgeway's osteogenic sarcoma and Gardner's lymphosarcoma than the compound VLB itself, which is the fundamental alkaloid from which these carboxamides are derived. Some of these amide derivatives have approximately the same effect as vincristine against the same tumors. One of these amides, 4-desacetyl-VLB-C-3-carboxamide or vindesine, is subjected to ongoing clinical experiments in humans, and it has been found that the compound is effective against certain leukemia occurrences. In humans, vindesine appears to have a lower degree of neuro-toxicity than vincristine, and the compound is clearly effective against vincristine-resistant forms of leukemia.

Belgian Patent Specification No. 813,168 and Danish Patent Specification No. 141,511 also mention, as mentioned, 4-desacetyl-VLB-C-3-carboxhydrazide as an effective anti-tumor agent 5 against transplanted tumors in mice. The compound has also been shown to be effective against Ridgeway's osteogenic sarcoma, Gardner's lymphosarcoma and P1534 (3) leukemia.

The above-mentioned Danish Patent Specification No. 141,511 discloses, in addition to 4-desacetyl-VLB-C-3-carboxhydrazide, several other compounds which are closely related to the compounds of the invention. These known compounds have similar effects to the compounds of the present invention, but as will be seen from the following test report, the presently mentioned derivatives of 4-desacetyl-VLB-C-3-carboxhydrazide are more active than the above known compounds. or they are effective at lower doses or become effective in less time.

In the present compounds of the general formula I, R may be methyl, ethyl, n-propyl, isopropyl, β-hydroxyethyl, β-acetoxyethyl or alkoxycarbonyl, especially ethoxycarbonyl or methoxycarbonyl.

The compounds of formula I are named as derivatives of 4-desacetyl-VLB-C-3-carboxhydrazide. A systematic naming of these compounds should include the term "3-descarbomethoxy", but this term is omitted as it is implicitly included in the term "C-3-carboxyhydrazide" because the C-3-carbomethoxy group in VLB is replaced.

30 Other naming systems can also be used. Eg.

For example, the compounds may be named as derivatives of 4-desacetyl-VLB-23-desmethoxy-23-hydrazide, referring here to the methoxyl group at C-23 being replaced by hydrazide. At times, it is preferred to name the compounds as derivatives of C-3-carboxhydrazide.

Hydrazine contains two nitrogen atoms, which in a hydrazide 1 2 5 are numbered as follows: C-N -N -H.

II I I

0 Η H

The hydrozide derivatives prepared according to the invention 2 are all N derivatives.

The 4-desacetyl-VLB-C-3-carboxazide used as starting material in the process of the invention can be prepared from 4-desacetyl-VLB-C-3-carboxhydrazide.

This compound can be prepared by reaction between hydrazine and VLB in accordance with the procedure described in Belgian Patent No. 813,168. According to this process, VLB and hydrazine are heated in a closed reaction vessel using anhydrous ethanol as the solvent. Alternatively, 4-desacetyl-VLB can be reacted with anhydrous hydrazine under the same conditions. The reaction between hydrazine and VLB serves to hydrolyze the acetoxy group at C-4, and the reaction product thereby becomes 4-desacetyl-VLB-C-3-carbohydride, regardless of whether VLB or 4-desacetyl-VLB are used as starting material. 4-Desacetyl-VLB-C-3-carboxhydrazide can then be converted to the corresponding C-3-carboxazide in accordance with the process described in the aforementioned Belgian Patent No. 813,168 in which the hydrazide is treated with nitrite in acidic solution. The 4-desacetyl-VLB-C-3-carb oxazide thus prepared can then be reacted with the various substituted hydrazines as claimed. 1 to give 30 of compounds of formula I. If reacted with N1- (8-hydroxyethyl) hydrazine to give the corresponding 4-desacetyl-VLB-C-3- [N - (β-hydroxy) ethyl] carboxyl Hydrazide, the corresponding β-acetoxyethylhydrazide derivative can be prepared therefrom by acetylation.

The invention is further illustrated by the following examples.

Preparation of Starting Materials 5 (1) 4-Desacetyl-VLB-C-3-carboxhydrazide 4-Desacetyl-VLB was heated in anhydrous ethanol with an excess of anhydrous hydrazine in a closed reaction vessel at about 60 ° C for about 18 hours. The reaction vessel was cooled and opened, the contents were removed and the volatiles were evaporated in vacuo. The resulting evaporation residue, consisting of 4-desacetyl-VLB-C-3-carboxhydrazide, was collected in methylene chloride and the methylene chloride solution washed with water. The methylene chloride phase was separated and dried and the methylene chloride removed by evaporation in vacuo. The residue was dissolved in a clear mixture of chloroform and benzene and chromatographed over silica gel. An eluent consisting of benzene, chloroform and triethylamine was used to develop the chromatogram. The first chromatographic graphene fractions contained unreacted 4-desacetyl VLB.

Subsequent fractions were found to contain 4-desacetyl-18'-descarbomethoxy-VLB-C-3-carboxhydrazide (previously described by Neuss et al., Tetrahedron Letters, 1968, 783). The following fractions, which by thin layer chromatography were found to contain 4-desacetyl-VLB-C-3-carboxhydrazide, were combined and the solvent was evaporated in vacuo. The resulting solid melted at about 219-222 ° C during decomposition.

(2) 4-Desacetyl-VLB-C-3-carboxazide A solution of 678 mg of 4-desacetyl-VLB-C-3-carboxhydrazide in 15 ml of anhydrous methanol was prepared. To this solution was added about 50 ml of 1 N hydrochloric acid and the resulting solution cooled to about 0 DC. Then about 140 mg of sodium nitrite was added and the resulting reaction mixture was stirred for 10 minutes while. the temperature was kept at about 0 ° C. By the addition of sodium nitrite, the solution was stained dark reddish brown. Then, the reaction mixture was made basic by adding an excess of cold 5¾ aqueous sodium bicarbonate.

The aqueous solution was extracted three times with methylene chloride to dissolve the 4-desacetyl VLB-C-3-carboxazide formed in the methylene chloride phase. can usually be used without further purification.

15 FINAL PRODUCTS

EXAMPLE 1 2

Preparation of 4-desacetyl-VLB-C-3-N-methylcarboxyl hydrazide

For a solution of approx. 1 g of 4-desacetyl-VLB-C-20 3-carboxazide in 150 ml of methylene chloride was added 20 ml of CH 2 NNNH 2. The reaction vessel was closed and left at room temperature for 6 hours. The methylene chloride solution was then extracted several times with water to remove the excess CH 2 NHNf 3 solution was dried, and the solvent was evaporated in vacuo. The resulting product (250 mg golden brown, amorphous solid) 2 was 4-desacetyl-VLB-C-3-N-methylcarboxyhydrazide having the following physical data: MS: m / e = 782 (M +), 441, 355, 154 IR: V = 3450 cm -1 (-NH) 1715 cm -1 (-C00CH3) 1655 cm -1 (-C00NH-) in Example 148 2

Preparation of 4-desacetyl-VLB-C-3-N, N -dimethylcarboxhydrazide

To a solution of about 1.5 g of 4-desacetyl-VLB-C-3-carboxazide in methylene chloride was added 20 ml of H 2 NNCCH 3. The reaction vessel was closed and left at room temperature for about 60 hours. The resulting solution was evaporated to dryness and the residue was dissolved in methylene chloride. The methylene chloride solution was extracted once with dilute ammonium hydroxide and then with water to remove excess 1 NNCCH 3. Then the methylene chloride solution was dried and evaporated to dryness. The resulting powder was chromatographed over silica gel and eluted with ethyl acetate and methanol (1: 1). The fractions which, by thin-layer chromatography (si lica eluting with 1: 1 ethyl acetate: methanol) were found to contain the desired product, were combined and the combined fractions were evaporated to dryness. The golden brown amorphous powder thus formed consisted of 4-des-2,220 acetyl-VLB-C-3-N, N -dimethylcarboxhydrazide having the following physical characteristics: Yield 250 mg.

MS: m / e r 796 (M +), 737, 455, 355, 154 I.R. : V = 1715 cm -1 (C00) 1670 cm -1 (CON) EXAMPLE 3 2

Preparation of 4-desacetyl-VLB-C-3-N - (ε-hydroxy) ethyl carboxhydrazide

About 3 g of 4-desacetyl-VLB-C-3-carboxazide was dissolved in methylene chloride and 15 ml of N 2 NH CH 2 CH 2 OH was added.

The reaction vessel was closed and the reaction mixture was stirred overnight at room temperature. The mixture was then evaporated and the residue was partitioned between chloroform and water. The chloroform solution was washed twice with water, dried and evaporated to dryness.

The amorphous powder was chromatographed over silica gel eluting with ethyl acetate and methanol (1: 1). The fractions which by thin layer chromatography (silica gel eluted with 1: 1 ethyl acetate: methanol) were found to contain the desired product were combined and the combined 10 fractions were evaporated to dryness. There was thus obtained 1 g of a golden brown amorphous powder having the following physical characteristics: I.R: Ϋ = 1720 cm -1 (C00) 1655 cm -1 (CON) EXAMPLE 4 2

Preparation of 4-desacetyl-VLB-C-3-N - (β-acetoxy) ethyl-carboxhydrazide 2 1624 mg of 4-desacetyl-VLB-C-3-N - (β-hydroxy) ethylcarboxhydrazide was dissolved in 50 ml methylene chloride. To this solution was added 220 mg of acetic anhydride. The reaction vessel was closed and left overnight at room temperature. The reaction mixture was washed with dilute ammonium hydroxide and then with water. The resulting methylene chloride solution was dried and evaporated to dryness, and the amorphous powder was chromatographed over silica gel. The chromatogram was eluted with ethyl acetate and methanol (1: 1). The fractions which by thin layer chromatography on silica gel eluting with ethyl acetate and methanol (1: 1) were found to contain the desired product were combined and the combined fractions were evaporated to dryness. There was thus obtained 215 mg of a golden brown amorphous powder consisting of 4-dec-2-acetyl-VLB-C-3-N - (ω-acetoxy) ethylcarboxhydrazide having the following physical characteristics: MS: m / e = 854 ( M +), 795, 651, 513, 355, 154 EXAMPLE 5 2 Preparation of 4-desacetyl-VLB-C-3-N-ethoxycarbonylhydrazide

About 900 mg of 4-desacetyl-VLB-C-3-carboxazide was dissolved in methylene chloride. To this solution was added 1 H 2 NNHCO 2 CO 2 dissolved in methylene chloride. Reaction vessel 010 was closed and left at room temperature overnight. Then, the reaction mixture was evaporated to dryness and the residue was partitioned between methylene chloride and water. The methylene chloride phase was washed twice with water, dried and evaporated to dryness. The resulting golden brown amorphous powder consisting of 4-desacetyl-VLB-C-3-N-ethoxycarbonylhydrazide was converted to the corresponding sulfuric acid salt by dissolving the amorphous powder in anhydrous ethanol and then adjusting the pH to about 4 2% sulfuric acid in absolute alcohol. This precipitated the sulfate, which was then collected by filtration and dried. The resulting product was a golden brown, amorphous powder (104 mg) whose melting point could not be determined.

The compounds of formula I inhibit the growth of 25 transplanted tumors in mice. The activity of the compounds of this invention was demonstrated by administering them to mice, usually by intraperitoneal route at a given dose for 7-10 days. The size of the tumor was measured after 3 or 7 days. The following Table I lists the results of the experiments performed, in which mice with transplanted tumors were successfully treated with a compound of formula I. Column 1 of the table gives the name of the compound studied, column 2 indicates the nature of the transplanted tumor, 5 column 3 indicates the dose level and number of days the dose was administered, and column 4 indicates the percentage inhibition of tumor growth (ROS is an abbreviation for Ridgeway's osteogenic sarcoma and GLS for Gardner's lymphosarcoma).

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A clinical trial of a compound of formula I may be performed in accordance with a procedure proposed by SK Carter in a section entitled "Study Design Principles for the Clinical Evaluation of Ne \ i / Drugs as Developed by the Chemotherapy Program of the National Cancer Institute ", found on pages 242-289 of book 20" The Design of Clinical Trials in Cancer Therapy ", edited by Maurice Staquet (Futura Publishing Co., New York, 1973).

Claims (3)

Patent Claims: 1 Nt1 OR wherein R is alkyl of 1-3 carbon atoms, β-hydroxyethyl, 5β-acetoxyethyl or alkoxycarbonyl of 1-3 carbon atoms and R 1 is hydrogen and, when R is alkyl of 1-3 C atoms, also methyl characterized by reacting a 4-desacetylvinylcaleucoblastin C-3 derivative of the formula: 148655 5 * QH r-r> -c2h5 11 * 8 * 1 xgj 3 > * sl8 * J 11 14 'f 1 ^ T ^ | “° -CH3 (IA) Η I &« 5χ I 1 (vN | 17 I 14 111 Il9 CH, -0 —1¾. JL 1 3 Yy-Y Wherein Q is N, in an inert solvent, with medν, R R1 wherein R is alkyl of 1-3 C atoms, β-hydroxyethyl, β-ace toxyethyl or alkoxycarbonyl having 1-3 C atoms in the alkoxy group and R 1 is hydrogen or, when R is alkyl having 5 1-3 C atoms, also methyl, optionally followed by an acetyl ring when R is β-hydroxyethyl. An analogous process for the preparation of derivatives of 4-desacetyl-vincaleucoblastin-C-3-carboxhydrazide of the general formula: 5 * OH 7 ---- C2H5 11 'S * I ) 14 »5 N 17 -C - 0-CH, II„ 3 Η ISI 8 10r-N j] 7 I 14 111 Jl9 I 6 - -ch2-ch3 CH, -0 — U JL1 OB 17 1 ch3 i / a C-NH-N. Analogous process according to claim 1 for preparation 2 of 4-desacetyl-vincaleucoblastin-C-3-N-methylcarboxyl hydrazide, characterized in that 10 4-desacetyl-vincaleucoblastin-C-3-carboxazide is reacted with methyl hydrazine. An analogous method according to claim 1 for the preparation of 2 4-desacetyl-vincaleucoblastin-C-3-N, N -dimethylearboxamide.