JPH0161118B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a method for producing a novel cytostatic compound and a pharmaceutical composition containing the compound. More specifically, the present invention relates to the production of novel compounds of the following general formula (). In the general formula (), R ⁴ is β-hydroxyl and R ³ is α-ethyl, or R ⁴ is hydrogen and R ³ is β-ethyl, and R″ is a carbon number other than ethyl. Straight chain alkyl having 1 to 10 carbon atoms or branched alkyl having 3 to 10 carbon atoms (in the straight chain alkyl and the branched alkyl,
the carbon atom bonded to the N _a —CH ₂ —O— group is a primary or secondary carbon atom), or aralkyl in which the alkyl moiety has 1 to 3 carbon atoms, R ¹ is methoxy, and R ² is acetyl. General formula () [wherein R ¹ , R ² , R ³ , R ⁴ and
R″ has the same meaning as previously defined] exhibits cytostatic activity. Compounds of general formula () have the same meaning as previously defined. General formula () or their acid addition salts by oxidation. In the above formula, R ¹ , R ² , R ³ , R ⁴ and R'' have the same meaning as defined before. This reaction essentially involves the formation of a group N _a - in a compound of general formula ().
Including changes in the structure of _CH3 . This change is based on N _a ―
Any bis-indole alkaloid including _CH3 may be used. The starting compounds of general formula () are known. For example, vinblastine [R ¹ = methoxy, R ² = acetyl, R ³ = α-ethyl and R ⁴ = β-hydroxyl] is disclosed in U.S. Pat. No. 3,097,137; Lieurocidin [R ¹ = methoxy, R ² = acetyl, R ³ = β-
For ethyl and R ⁴ = hydrogen] and its isolation
It is described by N. Neuss et al. [Tetrahedron Letters]
Letters) 1968 , 783], and its synthesis was reported by Potier et al. [J.Am.Chem.
Soc. 98 , 7017 (1976)]. Although the starting compound has cytostatic activity, it is more toxic than the compounds of general formula () prepared from this compound. The acute toxicity of the compounds according to the invention was
31 g male Swiss mice were tested. The test was performed on groups of 6 mice in each group. Test compounds were administered intraperitoneally as an injection solution prepared with saline and optionally an additional drop of Tween 80.
Doses were increased continuously from non-lethal to lethal doses. The results were evaluated by the Lichfield-Wilcoxon method and are reported in Table 1 below in comparison with vinblastine, vindesine and vincristine.

Table: According to the results listed in Table 1, the new compounds are 15-25 times less toxic than vincristine and vindesine, and 8-13 times less toxic than vinblastine. Unlike vincristine and vindesine, no paralytic side effects were observed with the new compound at doses up to _LD50 . The cytostatic activity of the new compounds according to the invention was also tested in tissue culture and various transplanted tumor lines. Test compounds were dissolved in tissue culture (HeLa cell culture) at concentrations from a dose threshold of 1 x 10 ^-3 μg/ml up to 100 μg/ml. In 24-hour cultures, intravital microscopic evaluation of metaphase inhibitory effects yielded the following results.

Table: Under similar conditions but using stained tissue cultures, relatively small differences were observed. This can be classified as follows: The first stage (minimum effective dose) is characterized by an increased proportion of blocked mitosis. Some of these are abnormal mitoses, eg three-group mitosis, or polar chromosomes also appear. Generally, the later stages have already disappeared. In the second stage, a strong metaphase block is observed and normal mitosis is almost absent. Chromosomes are in the shape of flaccid globules. The proportion of cells in interphase is low. In the third stage, the chromosomes clump together into a mass in the center of the cell. This condition is called "Ball-Metaphase" pycnomitosis. In the fourth stage, interphase cells are already affected. More specifically, blocked cells are unable to participate in mitosis and their number decreases. The cytoplasm of the cell (in interphase) is elongated,
The edges are irregular, "tufted" and the cells often have an elongated, fibroblast-like shape. Finally, in the fifth stage, the cytoplasm is filled with a fine network structure, and it is clear that the interphase cells have been destroyed by the treatment. The stages were well distinguishable when testing the compounds according to the invention. The following observations were made: The most effective compound was the heptoxy derivative, which caused a strong metaphase blockade already at a dose of 0.001 μg/ml. Pycnomytosis was observed at a dose of 0.1 μg/ml, and interphase cells were affected at doses of 1 to 10 μg/ml. The isobutoxy derivative had a lower degree of effect, ie, normal mitosis could still be observed even at a dose of 0.001 μg/ml. Benzyloxy derivatives have the weakest effect. At the lowest dose, this compound caused a minimal block with several types of modified mitosis, but no anaphase was observed. At 0.01 μg/ml, the blockade was moderate, and a dose of 1 μg/ml induced strong blockage and pycnomitosis. The effects of the new compounds on intraperitoneally implantable tumors (P388 murine leukemia and NK/Ly ascites lymphoma) are discussed below. BDF P388 Leukemia Test
This study was carried out on hybrid mice. The study was performed in groups of 6 mice each, with 10 ⁶ tumor cells implanted intraperitoneally per animal. After transplant
After 24 hours, test compound administration was started. Treatments were administered intraperitoneally, and the weight and condition of the animals were controlled daily. The effect obtained for the treated animals is expressed in % of the mean lifespan (in days) of the control group. The results, listed in the table below, show that the test compound significantly increases the longevity of mice bearing P388 leukemia.

【table】

[Table] *Animal died without tumor.
Swiss-H/Riob from our variety
-H/Riop) Groups of 10 outbred mice were implanted intraperitoneally with 5×10 ⁶ ascites tumor cells.
Treatment was started 24 hours after implantation and the compound was administered daily.
A total of 5 doses were administered. The average lifespan of the control group was 15.7
It was hot in the sun. N-desmethyl-N-(methoxymethyl)-
A similar test was performed with 20'-desoxy-lyurocidin. In these studies, dosage and number of treatments were varied. The results obtained are shown in Table 4.

[Table] Urocidin
4×7
17.1 12.1 142
8×1
21.7 12.2 178
8×0
.5 15.3 12.2 126

Claims (1)

[Claims] 1 General formula () [In the formula, R ⁴ is β-hydroxyl and R ³ is α-ethyl, or R ⁴ is hydrogen and R ³ is β-ethyl, and R″ is C _1-8 alkyl (however, ethyl ) or phenyl C _1-3 alkyl, and R ² is acetyl], the method comprises the steps of: [wherein R ¹ , R ² , R ³ and R ⁴ have the same meanings as defined above] or their acid addition salts and a large excess of the general formula () R″—OH ( ) [wherein R'' has the same meaning as defined above] in the presence of chromium trioxide, an organic solvent, acetic anhydride and an acid at a temperature of -60 to -30°C. Seshime: PH of the obtained reaction mixture is 8
10 and; after purification, isolating the product obtained. 2 Alcohol of the general formula () is converted to the alcohol of the general formula ()
The method according to claim 1, wherein the compound is used in a molar equivalent of 50 to 150. 3. The manufacturing method according to claim 2, wherein a chlorinated hydrocarbon is used as the organic solvent.