JPS6259117B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention provides a novel α・α-trehalose-6・
This invention relates to a 6'-medium chain fatty acid diester and an antitumor agent containing the same as an active ingredient. α·α-trehalose is a type of so-called disaccharide composed of two molecules of glucose, and is a compound widely distributed in nature. Various derivatives of this compound have also been synthesized and their pharmacological activities are being investigated. For example, it has been reported that mixtures of various diesters with different fatty acid bonding positions have antitumor effects. The present inventors have discovered that α・α-trehalose 6.
While synthesizing various 6'-fatty acid diesters and studying their pharmacological effects, we found the following formula () (In the formula, n represents an integer from 6 to 10) The α·α-trehalose-6·6′-medium chain fatty acid diester has excellent antitumor activity,
They found that it is useful as a medicine and completed the present invention. That is, an object of the present invention is to provide a novel α·α-trehalose-6·6′-medium chain fatty acid diester represented by the formula (). Another object of the present invention is to provide an antitumor agent containing α·α-trehalose-6·6′-medium chain fatty acid diester represented by the formula () as an active ingredient. The α·α-trehalose-6·6′-medium chain fatty acid diester (hereinafter simply referred to as trehalose medium chain fatty acid diester) () of the present invention can be obtained by converting α·α-trehalose () into 2·6′-medium chain fatty acid diester () according to the following reaction formula, for example.
3,2',3'-tetra-O-benzyl-α,α-trehalose (), and then reacted with medium chain fatty acid () or its active derivative to create a novel intermediate 2,3,2'・3'-tetra-O-benzyl-
It is produced by obtaining α·α-trehalose-6·6′-medium chain fatty acid diester (), which is further reduced and debenzylated using a reduction catalyst. (In the formula, Bn represents a benzyl group, and n has the meaning described above.) In the above method, 2.
3,2',3'-tetra-O-benzyl-α,α-trehalose is produced by reacting benzaldehyde with trehalose represented by the formula () using a conventional method. -Benzylidene-α・α
- trehalose, then converted to 2,3,2',3'-tetra-O-benzyl form with benzyl chloride,
It can be obtained by further reacting with an acid. In addition, the reaction between the compound () and the medium-chain fatty acid () or its active derivative is carried out according to general esterification reaction conditions, usually in the presence of a deoxidizing agent such as pyridine or triethylamine, for 1 mole of the compound (). , preferably by reacting medium chain fatty acids () or active derivatives thereof in a proportion of 1.8 to 2.4 moles. This reaction is preferably carried out at ~30° C. for 1 to 24 hours under ice cooling in the presence or absence of a solvent. Examples of the solvent used here include methylene chloride, chloroform, benzene, toluene, ether, tetrahydrofuran, and dioxane. The reductive debenzyl reaction of the compound () thus obtained is carried out by catalytic reduction using preferably 0.5 to 10 moles of a reduction catalyst per 1 mole of the compound () in an appropriate solvent. Examples of the reduction catalyst include palladium black, palladium-carbon, and Raney nickel. Also,
Examples of the solvent used here include alcohols such as methanol, ethanol, and isopropanol; halides such as chloroform and methylene chloride; and mixed solvents thereof. Next, the pharmacological action of the trehalose medium chain fatty acid diester () of the present invention will be shown. (1) Cell killing effect of trehalose medium-chain fatty acid diester: Using Ryuchemia L-5178Y cells,
10 ⁵ in PRMI1640 medium containing 10% fetal bovine serum
The sample was added to this medium at a predetermined concentration. After culturing in a 5% CO ₂ incubator at 37°C for 48 hours, the number of living cells was counted under a microscope, and the proliferation rate was determined by comparing it with the control group, and plotted on logarithmic probability paper. The IC ₅₀ value was determined. Test concentrations are 25, 50,
The concentrations were 100, 200 and 400 μg/ml. The results are shown in Table 1.

[Table] (2) Survival prolonging effect of trehalose medium-chain fatty acid diester on Ehrlichi tumor-bearing mice: ICR-JCL male mice (5 to 6 weeks old) Ehrlichi cancer cells grown intraperitoneally for 7 days were collected, and then treated with physiological saline. After washing twice with
The cells were adjusted to ×10 ⁵ cells/ml, and 0.5 ml of the cells was intraperitoneally transplanted into ddY male mice (6 to 7 weeks old).
Groups of 8 to 10 ddY mice were used after tumor implantation.
From 24 hours onwards, the sample was intraperitoneally administered once a day for 7 consecutive days. Efficacy was determined by observing whether the tumor was alive or dead for 60 days after tumor transplantation, and calculating the survival rate from the number of survival days using the formula below. Life extension rate (ILS%) = Survival days of drug administration group - Survival days of control group / Average survival days of control group x 100 The sample is suspended in 0.5% CMC physiological saline, and the administered volume is 0.1 ml per 10 g of mouse body weight. The sample concentration was adjusted as follows. The results are shown in Table 2.

【table】

[Table] (3) Meth of trehalose medium chain fatty acid diester
- Engraftment inhibitory effect on A cells: Meth-A cells proliferated in the peritoneal cavity of BALB/C male mice (6-7 weeks old) were collected, washed twice with physiological saline, and then collected at 9 x 10 ⁵ cells/ml. Adjusted to. On the other hand, the sample was prepared with 0.5% CMC saline.
2. Suspended at 1 mg/ml. Immediately before administration, mix the cells and sample suspension at a ratio of 1:1 (V/V),
0.1 ml of the solution was subcutaneously inoculated on the back of BALB/C mice. There were 5 animals in each group, and the number of Meth-A cells engrafted and tumor weight were measured 17 days after transplantation. The results are shown in Table 3.

[Table] From Table 3, all of the five animals in the control group contained Meth.
-A cells were engrafted. On the other hand, in the mixed inoculation group of 8 compounds of the present invention, 0, 2, and 5 out of 5 mice were prevented from engrafting Meth-A cells at 0.5, 1.0, and 2.0 mg, respectively. Furthermore, the tumor weights on day 17 were lower in all groups administered with compound 8 of the present invention than in the control group. As mentioned above, trehalose medium chain fatty acid diester has an antitumor effect, and its effect is as follows:
It is significantly stronger than the corresponding compounds with 16 to 22 carbon atoms. The toxicity of the trehalose medium-chain fatty acid diester of the present invention is due to the fact that, for example, sucrose fatty acid esters, which are similar substances, are designated as food additives and are widely used as harmless surfactants in foods, medicines, cosmetics, etc. As is clear from the fact that it is a safe compound with almost no toxicity. In addition, the dosage forms for administering the compounds of the present invention include various dosage forms depending on the dosage form, such as oral and parenteral dosage forms, such as oral dosage forms such as tablets, capsules, powders, granules, and liquids; subcutaneous, It can be administered parenterally, such as intramuscular or intravenous injections, annular fluid mixtures, or suppositories. The above formulation can be performed by a method known per se. That is, trehalose medium chain fatty acid diester with excipients such as starch, lactose, and mannitol;
Binders such as sodium carboxymethylcellulose and hydroxypropylcellulose; disintegrants such as crystalline cellulose and calcium carboxymethylcellulose; lubricants such as talc and magnesium stearate; fluidity improvers such as soft hydrosilicic acid, etc., are combined as appropriate. Tablets, capsules, powders, or granules can be produced by this process.
In addition, solutions and injections are manufactured by dissolving trehalose medium-chain fatty acid diester in vegetable oil, etc. to make an oil-based injection, or by dissolving or suspending it in water, etc. using a conventional method to make a syrup, etc. be able to. Furthermore, for making suppositories, commonly used bases,
For example, it can be produced by dispersing it in cacao butter, synthetic oil, etc. by a conventional method and then solidifying it. The dosage of the trehalose medium chain fatty acid diester of the present invention varies depending on the degree of the disease, but
Usually 0.1 to 2000 for oral administration in adults
mg/Kg, and in the case of parenteral administration, it is preferable to administer 0.05 to 1000 mg/Kg once a day or divided into several times a day. Next, the present invention will be explained with reference to Examples. Example 1 2.3.2'.3'-tetra-O-benzyl-α.
20 ml of pyridine and 20 ml of dichloromethane were added to 2.11 g of α-trehalose to dissolve it, and 1.26 g of decanoyl chloride was added dropwise thereto under stirring at 0° C. under ice cooling. After stirring for 30 minutes, the mixture was returned to room temperature and further stirred for 1 hour. The resulting reaction mixture was poured into ice water, extracted with chloroform, and then chloroform was distilled off under reduced pressure. The residue was purified by silica gel column chromatography to give 2,3,2',
2.79 g of 3'-tetra-O-benzyl-6,6'-di-O-decanoyl-α,α-trehalose (yield: 92
%) was obtained. Example 2 2,3,2',3'-tetra-O- obtained in Example 1
Benzyl-6・6′-di-O-decanoyl-α・α
- Dissolve 2.67 g of trehalose in 40 ml of chloroform and add 1.00 g of palladium black as a catalyst to this solution.
g was added, hydrogen gas was introduced, and catalytic reduction was carried out for 1 hour. The resulting reaction solution was filtered to remove the catalyst,
The liquid was concentrated to dryness under reduced pressure. The residue was recrystallized from isopropanol to give 6,6'-
Di-O-decanoyl-α・α-trehalose 1.06
g (yield 62%). Example 3 The compounds shown in Table 4 were obtained in the same manner as in Example 1. At the same time, the compound obtained in Example 1 is also shown in the table.

[Table] Example 4 In the same manner as in Example 2, the compounds shown in Table 5 were obtained. At the same time, the compound obtained in Example 2 is also shown in the table.

[Table] Example 5 Tablet One tablet having the following ingredients and quantities was manufactured by a conventional method. Trehalose medium chain fatty acid diester (decanoate) 100mg D-mannitol 150mg Crystalline cellulose 50mg Starch 28mg Carboxymethyl cellulose calcium 16mg Talc 4mg Magnesium stearate 2mg Total amount 350mg Example 6 Capsules Granules with the following ingredients and amounts were manufactured by a conventional method,
This was filled into one No. 3 capsule. Trehalose fatty acid diester (caprylate)
25 mg Crystalline cellulose 17 mg Light anhydrous silicic acid 7 mg Magnesium stearate 1 mg Example 7 Injection One injection was manufactured from the following ingredients and quantities in a conventional manner. Trehalose fatty acid diester (decanoate)
Add 25mg of olive oil to make a total volume of 1ml. Example 8 Suppositories After melting and stirring the following ingredients and amounts using a conventional method,
The mixture was molded and solidified to produce one suppository. Trehalose fatty acid diester (laurate)
100mg Cocoa butter 1100mg Total amount 1200mg

Claims (1)

[Claims] 1st-order equation () (In the formula, n represents an integer of 6 to 10.) α·α-trehalose-6·6′-medium chain fatty acid diester. 2nd order formula () (wherein n represents an integer of 6 to 10) An antitumor agent containing α·α-trehalose-6·6′-medium chain fatty acid diester as an active ingredient.