JPS61233649A - Novel compound having immunological activating action and production thereof - Google Patents

Abstract

NEW MATERIAL:3-Hydroxy-4-(hydroxymethyl)benzoic acid expressed by formula I and a salt thereof. USE:A medicine, which is a substance, having immunological activating action without cytotoxicity, and useful for immunological deficiency disease of an organisms with cancer, immunization and chemotherapy of cancer and prevention of metastasis. PREPARATION:An ester expressed by formula II (R<1> is alkyl; R<2> is H or lower alkyl) is reduced in the presence of a reducing agent, e.g. NaBH4, in a solvent, e.g. absolute methanol, at room temperature- while heating, etc. and the residual ester groups are hydrolyzed in the presence of an alkali (earth) metal hydroxide, e.g. NaOH, in an organic solvent, e.g. methanol, at room temperature- while heating to afford the aimed compound expressed by formula I.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a novel compound having immunostimulatory activity and a method for producing the same.

The present inventors succeeded in producing a compound represented by structural formula (1), a pharmacologically acceptable salt thereof, and a hydrate of the compound or a salt thereof.

That is, the gist of the present invention is a novel compound represented by the structural formula, a pharmacologically acceptable salt, and a hydrate of the compound or a salt thereof.

Next, a method for producing the compound of the present invention will be described.

The compound of the present invention is a compound represented by the general formula (II) (wherein R1 represents a straight-chain or branched alkyl group, and R2 represents a hydrogen atom or a straight-chain or branched lower alkyl group), which has a hydroxyl group. It is obtained by selectively reducing the carboxylic acid ester adjacent to , to a hydroxymethyl group, and further hydrolyzing it.

Representative examples of this raw material compound (IT) include hydroxyterephthalic acid dimethyl ester and hydroxyterephthalic acid diethyl ester.

In the reduction reaction, the raw material compound (rl) is subjected to appropriate reaction conditions.
Sodium borohydride, sodium dihydrobis-
(2-methoxyethoxy, 67) can be carried out using metal hydrides.

Typical examples of this reaction include, for example, when sodium borohydride is used as the reducing agent, a lower alkanol such as methanol or ethanol is used as the solvent,
Examples include a reaction carried out at room temperature or under heating, and a reaction carried out at room temperature or when sodium dihydrobis-(2-methoxyethoxy)aluminate is used as a reducing agent.

The hydrolysis reaction can be carried out using any known and commonly used method, but preferably in a mixed solvent of water and an organic solvent that is miscible with water such as methanol, or in a mixed solvent such as sodium hydroxide or barium hydroxide. It is carried out using an alkali metal or alkaline earth metal hydroxide at room temperature or under heating.

When a compound in which R1 is a linear or branched lower alkyl group and R2 is a hydrogen atom in the general formula (IT) is used as a raw material compound, the lead metal hydride must be reacted under appropriate conditions. It is obtained by

When a compound in which R' and R'' are both linear or branched lower alkyl groups in general formula (II) is used as a raw material compound, first, lead metal hydride is reacted under severe conditions to selectively react the compound. It is obtained by reducing the ester adjacent to the hydroxyl group to a hydroxymethyl group and then hydrolyzing the remaining ester to a carboxylic acid.

Further, as a result of examining the usefulness of the compound of the present invention (BF-127 substance) as a medicine, it has been found that the compound of the present invention has an immunostimulatory effect based on the following pharmacological action.

Written by Pujjlj - Rule 1) Effect on cell-mediated immunity The effect of BF-127 substance on cell-mediated immunity was investigated by inoculating mouse paws with sheep red blood cells as an antigen and immunizing them with delayed hypersensitivity (Del. .ayed-type
The effect of hypersensitivity (abbreviated as DTII) was examined as an index. The DTII reaction was performed by Lagrange et al. (P.ll.

Lagrange, G. B. Mackaness an
dT. E. Old 11er:. 1. Exp.

Med. ] 39 1529-1539.1974).

IO'' of sheep red blood cells were suspended in 0.05 me physiological saline and immunized by subcutaneously inoculating one heel of an ICR female 6-week-old mouse, and each group of BP-127 was administered at 0.25 +
n (2) was dissolved in saline and injected intraperitoneally. Four days later, 10" sheep red blood cells were subcutaneously injected into the heel of the other foot to induce a DTI+ response, and 24 hours later, that foot (Table 1) Effect of BF-127 on the establishment of delayed hypersensitivity in mice to sheep red blood cells Mouse: ICR (Note 1) Subcutaneous injection (Note 2) Intraperitoneal injection Footprint thickness of non-administered mice Table 1 As shown, administration of 1 mg ~ O, 17 Ig/mouse of BF-127 substance enhanced the degree of DT11 response by 58% to 134% compared to the control. Administration, and intravenous and subcutaneous injections are also seen; examples of oral administration are shown below.

ICR female mice, 6 weeks old, were immunized with sheep red blood cells as described above, and 1 mg, 1 (10 μg, and 10 μg of BF-127 substance) were each dissolved in 0.25 m of physiological saline and administered orally at the same time as immunization. were administered to examine its effects.

(Table 2) (Table 2) BF-127 by oral administration and intraperitoneal injection
Effect of Substance on Establishment of Delayed Hypersensitivity in Mice Footprint Thickness of Untreated Mice The results are shown in Table 2. Oral administration of the BF-127 substance enhanced the DT11 response by 30% to 50% at any dose. The cell-mediated immunity enhancing effect of the BF-127 substance was also observed in other strains of mice of different ages.

CDF, (DBA/2XBALB/CF, ) male,
Nine-week-old mice were immunized as described above and BF-1
Each of the 27 substances was administered intraperitoneally or orally and the results were observed. (Table 3) (Table 3) CDF, Effect of BP-127 on the development of delayed hypersensitivity to sheep red blood cells in mice CDF, mouse, male, heel thickness of 9-week-old untreated mice The results are shown in Table 3 As shown in Figure 3, the enhancing effect was observed in both intraperitoneal injection and oral administration. As described above, it has been revealed that the substance BP-127 acts to enhance cellular immunity, and this suggests that it can be used for immunochemotherapy of cancer.

Below, the effect of BF-127 substance on animal transplanted tumors was investigated.

2) Effect on animal transplanted tumor a) Effect on Gardner Lymphoma Gardner I lymphoma cells 1 transplanted once a week into female 100 week old C3H/He mice
05 were subcutaneously transplanted into the lumbar region of 5 syngeneic mice per group,
After 7 days, take 10 BF-127 substances once a day for 5 days.
μg/mouse, 1 μg/mouse, and 0.1 μg/mouse were intraperitoneally injected, and 30 days after tumor implantation, the tumor was excised and its weight was measured to determine the effect. (Table 4) (Table 4
) BP- for Gardner Lymphoma
127 effective tumor mean severity IF'.

(mg) (Note 3) (%) 0゛Cell transplantation Untreated mouse 1310-BF
-12710μg570 56.51μg
930 29.00.1μg950 27.5 BLM (Note 2) 100μg 760
42.0 (Note 1) Once a day from the 7th day after tumor implantation, 5
Daily intraperitoneal administration (Note 2) Pleomycin (Note 3) Measurement on day 300 after tumor implantation (average of 5 C311/He mice) As a result, administration of 10 μg/mouse of BF-127 substance caused tumor growth compared to control. Suppresses proliferation with an inhibition rate of 56.5
%Met.

(mouth) Effect on IMC Carcjnoma IMCC
Arcinoma is CDF, a tumor that is a naturally occurring ascites-type tumor in mice and transplanted into syngeneic mice at the Institute of Microbial Chemistry. Pathological examination revealed that this tumor is a ``1 skin undifferentiated tumor.'' It is being done. IM
5 x 10' C tumor cells were transplanted subcutaneously into the sebaceous area of 5 CDF mice per group, and 8 days later, 100μ of BF-127 was applied.
g ~ 1 μg/mouse was administered once a day for 5 days, and 30 days after tumor implantation, the tumor was excised and its weight was measured to determine the effect. At the same time, the anticancer substances mitomycin C 20 g/mouse and pleomycin 100 tp/mouse were used as controls (Table 5-A).

(Table 5) (A) Effect of BP-127 substance on IMC cancer (Note 1
) Intraperitoneal injection once a day for 5 days from 8 days after tumor implantation (Note 2) Pleomycin (Note 3) Mitomycin C (Note 4) Measurement on the 300th day after tumor implantation (average of 5 animals per group)
10μg~1 of BF-127 substance as shown in Table 5-A
Administration of μg/mouse showed similar inhibitory effects as the anticancer agents mitomycin C and pleomycin.

In addition, 1 × 10 cells were transplanted, and BP-127
The effects of the substance were examined by intraperitoneal injection or intraperitoneal injection of ~1 μg/1 μg (Table 5-B).

(B) (Note 1) Intraperitoneal administration once a day for 5 days from the 8th day after tumor implantation (Note 2) Oral administration once a day for 5 days from the 8th day after tumor implantation (Note 3) Tumor Measurement 300 days after transplantation (average of 5 animals per group)
As shown in Table 5-B, intraperitoneal injection and oral administration showed even better suppressive effects.

Since the BF-127 substance is a non-cytotoxic substance, the inhibitory effect of this substance on various tumors is thought to be due to a reaction mediated by the host. From the above experimental examples, BF-1
It is clear that the 27 substances are useful for immunodeficiency in cancer-bearing organisms, cancer immunity and chemotherapy, and prevention of metastasis.

Next, the present invention will be explained in more detail with reference to Examples, but the present invention is not limited thereto.

Examples Example 1 580 mg of hydroxyterephthalic acid dimethyl ester was dissolved in 30 mQ of anhydrous methanol, and 2.5 g of sodium borohydride was slowly added. After the addition was completed, the mixture was stirred at room temperature for 1 hour, and then purified with 6N hydrochloric acid under water cooling.
Adjusted to H1.95. The residue obtained by extraction with butanol and distillation of the solvent was purified by silica gel column chromatography (benzene-ethyl acetate, 9:1 to 4:1), and 3-hydroxy-4-(hydroxymethyl)benzoin was extracted. 418 mg of acid methyl ester was obtained. Yield 83%.

Crystallization from benzene-methanol-ethyl ether gave 320 mg of colorless crystals.

Melting point T (14.5-105℃ I Rv (KBr): 3430,320F1.2
920, 1708, 1695, 1612°1590.1
517, 1440, 1420, 1370, 1360, 1
310,1295°1280.1255. +235.1
220,1190.1178.1110,1095°+
030.978.950.923.880.872.8
40.805°790, 756cffl NMR (60MIlz, heavy methanol solution) δ:
3.88 (311,s), 4.72 (211,s),
7.3-7.7 (311) Example 2 316 ung of 3-hydroxy-4-(hydroxymethyl)benzoic acid methyl ester obtained in Example 1 was mixed with methanol] mf! 5me of 1N aqueous sodium hydroxide solution was added, and the saponification reaction was carried out at room temperature for 1.5 hours. After the reaction, the mixture was diluted with water, 5me of 1N hydrochloric acid was added, and extracted with ethyl acetate. Ethyl acetate was distilled off under reduced pressure.
-Hydroxy-4-(hydroxymethyl)benzoic acid 27
2 mg was obtained. Yield 93%. Crystallization was performed from a benzene-ethanol mixture to obtain 154 mg of colorless crystals.

Claims (2)

[Claims] (1) Compounds represented by structural formula (I) ▲Mathematical formulas, chemical formulas, tables, etc.▼, pharmacologically acceptable salts thereof, and hydrates of the compounds or salts thereof. (2) General formula (II) ▲There are mathematical formulas, chemical formulas, tables, etc.▼ (In the formula, R^2 represents a linear or branched alkyl group,
Furthermore, the structural formula (I) ▲ is characterized by reducing the ester represented by R^2 (representing a hydrogen atom or a linear or branched lower alkyl group), and then, if necessary, hydrolyzing the remaining ester group. There are mathematical formulas, chemical formulas, tables, etc. ▼ Method for producing the compound represented by.