DK148510B - Analogy process for preparing derivatives of 4- deacetylvincaleucoblastin C-3-carbohydrazide - Google Patents

Description

in 148510

The present invention relates to an analogous process for the preparation of novel derivatives of 4-desacetylvincaleucoblastin C-3-carboxhydrazide having the general formula Ij as set forth in claim 1 and the process of 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 diseases in 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 U.S. Patent No. 811,110), Leurosidine (Vinrosedine), and Leurocristine, hereinafter referred to as Wine Ristin (both described in U.S. Patent No. 3,205,220), Deoxy-VLB "A" and "B" (described in Tetrahedron Letters, 783 (1958), 4-desacetoxyvinblastine (described in U.S. Pat.

3,954,773), 4-desacetoxy-3'-hydroxyvinblastine (disclosed in U.S. Patent No. 3,944,554), leuroclobin (disclosed in U.S. Pat. No. 3,890,325), and vincadioline (disclosed in U.S. Patent No. 3,887,555 ). Two of these alkaloids, VLB and vineristin, are now marketed as medications for the treatment of certain diseases, especially leukemia-25 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 molecular structures involved are extremely complicated, which is why it is difficult to conduct chemical reactions that alter a specific functional group in the molecule without affecting other groups. Secondly, from Vinca rosea dimeric alkaloids which did not possess the desired chemotherapeutic properties have been prepared, 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. as they often differ only from the stereochemistry in a single place. 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. Successful attempts to alter physiologically active alkaloids include the preparation of 6,7-dihydro-VLB (described in U.S. Patent No. 3,332,868) and the replacement of the acetyl group at the higher alkanoyl carbon atom # 4 of the VLB ring system. groups or with unrelated acyl groups (see U.S. Patent No. 3,392,173). Several of these C-4 derivatives are capable of prolonging the life of mice inoculated with P1534 leukemia. C-3-carboxamide and C-3-carboxhydrazide derivatives of VLB, vinchristine, vincadiolin 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 specification). no. 813 168 and Danish presentation no. 141 511). These compounds are extremely interesting, as e.g. The 3-carboxamides of VLB are more effective against Ridgeway's osteogenic sarcoma and Gardner's lymphosarcoma than the compound VLB itself, which is the basic alkaloid from which these carboxamides are derived. Some of these amide derivatives have approximately the same effect as vineristin against the same tumors. One of these amides, 4-desacetyl-VLB-C-3-carboxamide or vindesin, is continuously subjected to 30 clinical experiments in humans, and the compound has been found to be effective against certain leukemia occurrences. In humans, vindinsine appears to have a lower degree of neurotoxicity than vineristin, and the compound is clearly effective against vineristin-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 against transplanted tumors in mice. The connection has also proved to be effective

Ridgeway's osteogenic sarcoma. Gardner's lymphosarcoma 5 and P1534 (J) 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 10 known compounds have similar effects to the compounds of the present invention, but as will be apparent from the subsequent experimental report, the presently disclosed derivatives of 4-desacetyl-VLB-C-3-carboxhydrazide are more active than the above-mentioned compounds. 15 effects, or they are effective at lower doses or become effective in a shorter time.

In the compounds of the general formula I, R may be acetyl, propionyl, n-butyryl, isobutyryl or benzoyl.

20

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.

Other naming systems can also be used. Eg. For example, the compounds can 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. In preference to I-30, it is preferred to name the compounds as derivatives of C-3-carboxhydrazide.

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

II I I

0 Η H

4 148510

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

The 4-desacetyl-VLB-C-3-carboxhydrazide 5 used as starting material in the process of the invention can be prepared by reaction between hydrazide and VLB according to the process 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-des-acetyl-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-carboxhydrazide, regardless of whether VLB or 4-desacetyl-VLB are used as starting material.

The invention is further illustrated by the following examples,

Preparation of Starting Material 20 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 evaporation residue was dissolved in a 1: 1 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 5 chromatographic fractions contained unreacted 4-desacetyl VLB. Subsequent fractions were found to contain 4-desacetyl-18'-descarbo-methoxy-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 the 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.

END PRODUCTS

EXAMPLE 1 2

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

To a solution of 1500 mg of 4-desacetyl-VLB-C-3-carboxyhydrazide in 50 ml of methylene chloride was added 300 mg of acetic anhydride. The reaction mixture was allowed to stand for 3 hours at room temperature, after which the methylene chloride solution was washed with dilute ammonium hydroxide and then with water.

The methylene chloride solution was dried and evaporated to dryness. The resulting golden brown amorphous powder consisted of 4-desacetyl-VLB-C-3-N-acetylcarboxhydrazide with the following physical characteristics: Yield = 750 mg.

MS: m / e = 810 (M +) I.R .: V = 3410 cm-1 (N-H) 1720 cm-1 (C00) 1670 cm-1 (CON)

Preparation of 4-desacetoxy-1 / LB-C-3-N-butryrylcarboxhydrazide Example 2 o

To a solution of 768 mg of 4-desacetyl-VLB-C-3-carbohydride in 50 ml of methylene chloride was added 120 mg of butyric anhydride. The reaction vessel was closed and left overnight at room temperature. Then, the reaction mixture was extracted with dilute ammonium hydroxide and the extract was poured off. The remaining methylene chloride solution was dried and evaporated to dryness. The residue was chromatographed over silica gel eluting with ethyl acetate and methanol (1: 1). The fractions which by thin layer chromatography were found to contain the desired product were combined and the combined fractions were evaporated to dryness. There was obtained 128 mg of a golden-brown amorphous powder consisting of 4-desacetyl-VLB-C-3-N-butryrylcarboxhydrazide with the following physical data: MS: m / e = 838 (M +), 497, 355, EXAMPLE 3 Preparation of 4-desacetyl - [LB-C-3-N-benzoylcarboxylic acid]

To a solution of 768 mg of 4-desacetyl-1 / LB-C-3-carboxy-hydrazide in 50 ml of methylene chloride was added 240 mg of benzoic anhydride. The reaction vessel was closed and set at 25 vec room temperature overnight. Then, the vessel was opened and the methylene chloride solution was washed with dilute ammonium hydroxide and then with water to remove formed benzoates. The methylene chloride solution was then 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 and 1: 1 ethyl acetate: methanol) were found to contain the desired product were combined and evaporated to dryness. There was thus obtained 150 mg of a tan amorphous powder consisting of 4-desacetyl-VLB-2 C-3-N-benzoylcarboxhydrazide with the following physical data: MS: m / e = 872 (M +), 813, 531, 355, 154 NM NMR: In accordance with proposed structure; new aromatic protons at ^ 7.3-8.0.

The compounds of formula I inhibit the growth of transplanted tumors in mice and / or extend the life of mice inoculated with tumors. 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 in cases where the administered compound inhibited the growth of the tumor. In the 15 cases where the life-prolonging effect was examined, the extra life of the treated animals was determined relative to the control animals. 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 under study, column 2 indicates the nature of the transplanted tumor. Column 3 indicates the dose, level, and number of days the dosing was administered, and Column 4 indicates the percent inhibition of tumor growth or the prolongation of survival time (GLS is an abbreviation for Gardner's lymphosarcoma, and B16 for a melanoma malignum ).

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When the hydrazide derivatives of Formula I are used as anti-neoplastic agents, the parenteral route of administration is used. For this purpose, isotonic solutions containing 1-10 mg per ml are used. ml of a compound of formula I.

The compounds are administered in an amount of between 0.01 and 1 mg per day. and preferably between 0.1 and 1 mg per kg of body weight. kg of body weight 1 or 2 times a week or 1 or 2 times every 2 weeks depending on both the activity and the toxicity of the compound concerned. Another method 10 for determining the therapeutic dose is based on the area of the patient's body surface, using an amount of between 0.1 and 10 mg per day. square meters of body surface every 7 or 14 days.

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 New 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: An analogous process for the preparation of derivatives of 4-desacetylvincaleucoblastin-C-3-carboxhydrazide of the general formula: 5 'OH 7 ---- C2H5 11' 8 * ηηι 3 «12g, \ V 11 (i) ιϊ \ itN ^ -co-ch, (JI 1 11 ^ H i O i 8 I lOr-- N 'i 14 11 Jl9 J 6' | 5 | ~ ~ CH2 "CH3 CH.-0 - ^ J ^ 1 J \ 3 / ~ 0H 3 ^ Vl8N N 'HO ^ r 17. In "CH, I C-NH-N ix" wherein R is alkanoyl of 2-4 C atoms or benzoyl, characterized by reacting a 4-desacety 1-vincaleucoblastin C-3 derivative of the formula: 148510 5 'OH ~ C2H5 1111 81 τα {31 12. g, \ 13'% ^^ i ^ \ i8 * / 11 14. N 17 ^ Y ^ -0-CH3 (ia) H I 0 I 8 v \ 7 j 10 | -N || (I 14 111 119 3 6 - -ch2-ch3 CH3 '° 17 I 1 CH3 j ^ C-Q * H. 0 wherein Q 1 is NHNH 4 with the acid anhydride or acid chloride of an alkanoic acid having 2-4 C atoms or benzoic acid. Analogous process according to claim 1 for preparation 2 of 4-desacetyl-vincaleucoblastin-C-3-N-acetylcarbox-5-hydrazide, characterized in that 4-desacetyl-vincaleucoblastin-C-3-carboxyhydrazide is reacted with acetic anhydride. Analogous process according to claim 1 for preparation 2 of 4-desacetyl-vincaleucoblastin-C-3-N-butryrylcarbox-hydrazide, characterized in that 4-desacetyl-vincaleucoblastin-C-3-carboxyhydrazide is reacted with butyric anhydride.