JPH06298764A - Immunosuppressant - Google Patents

Abstract

PURPOSE:To obtain an immunosuppressant useful as a therapeutic agent for autoimmune disease, allergic disease, transplantation of organs, etc., comprising radicicol as an active ingredient. CONSTITUTION:An immunosuppressant comprises radicicol of the formula as an active ingredient. Radicicol of the formula is obtained by culturing a filamentous fungus belonging to the genus Penicillium in a medium, by a conventional method, forming and accumulating radicicol in the culture mixture, purifying and isolating from the culture mixture and made into tablet, capsule, suppository, drop, etc.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a novel immunosuppressant. The immunosuppressive agent of the present invention is useful for treating autoimmune diseases, allergic diseases, organ transplants and the like.

[0002]

Radicicol, which is a microbial metabolite, is conventionally known to have an antifungal action and an anticancer action (Nature, 171, 344 (1953) and Neoplasma). , 24, 21 (1977)]. Further, it is known that a radicicol derivative obtained by modifying a phenolic hydroxyl group with various acyl groups has an antitumor effect (Japanese Patent Laid-Open No. 4-226991).

[0003]

[Problems to be Solved by the Invention] As immunosuppressants, adrenocortical hormones, antimetabolites, alkylating agents, alkaloids, antibiotics, anti-lymphocyte globulin, anti-CD3 monoclonal antibodies, etc. are known. It is used as a therapeutic drug for allergic diseases and organ transplants. However, they are not always satisfactory in terms of efficacy, sustainability, side effects, etc., and there is a demand for the development of even more excellent immunosuppressive agents.

[0004]

According to the invention, the formula (I)

[0005]

[Chemical 2]

An immunosuppressive agent containing radicicol represented by the formula as an active ingredient is provided. Radicicol is Penicill
It is obtained by culturing a filamentous fungus belonging to the genus ium in a medium, producing and accumulating in the culture, and purifying and isolating from the culture [Journal of Natural Products (J. N.
atural Products), 42 , 374 (1979)]. Next, the immunosuppressive action of radicicol will be described in Test Examples.

Test Example T Cell Proliferation Inhibition Test in Mouse Lymphocyte Mixed Reaction Spleens were aseptically removed from AKR mice [Japan SLC, Inc.] to prepare single cell suspensions. Mitomycin C (MMC) [Kyowa Hakko Kogyo Co., Ltd.] was added to this suspension (final concentration 50 μg / m
l) and incubated at 37 ° C for 30 minutes. After culturing, the cells were washed 3 times with a solution (HBSS) containing 2.5% fetal bovine serum (FCS, Gibco) in Hanks' balanced salt solution (Gibco), and 1x10 ⁷ cel.
Adjusted to ls / ml.

[0008] In each well of a 96-well microtiter plate, B10.BR mouse [Nippon SLC, Inc.] 50 μl lymph node cell suspension (containing 1.5 × 10 ⁵ cells) and AKR mouse spleen cell suspension 50 μl 100 μl of radicicol solution (containing 5 × 10 ⁵ cells) and each test concentration was added, and the mixture was cultured in a CO ₂ incubator at 37 ° C. for 72 hours. 18 hours before the end of culture [ ³ H]
-Thymidine 1.0 μCi was added. After completion of the culture, the cells were collected on a filter paper with a cell harvester, dried, and a toluene scintillator was added, and the amount of [ ³ H] -thymidine incorporated into the cells was measured with a liquid scintillation counter (test group).

As a control group, the radicicol solution was not added, and the culture was carried out in the same manner as above to measure the radioactivity of [ ³ H] -thymidine incorporated into the cells. The T cell proliferation inhibition rate was calculated according to the following formula.

[0010]

[Equation 1]

The results are shown in Table 1.

[0012]

[Table 1]

According to Table 1, radicicol suppressed T cell proliferation in mouse lymphocyte mixed reaction and showed a clear immunosuppressive action. The acute toxicity (LD50) of radicicol was 175 mg / kg when administered intravenously to rats [CRC Handbook of Antibiotic C
Ompounds), 2 , 448 (1980)], has low toxicity and can be safely used in a wide dose range.

Radicicol has an excellent immunosuppressive action and is useful as a therapeutic agent for autoimmune diseases, allergic diseases, organ transplants and the like. Radicicol is orally or parenterally administered as it is or as various pharmaceutical compositions. Examples of the dosage form of such a pharmaceutical composition include tablets, pills, powders, granules, capsules, suppositories, injections, and drip infusions.

For the formulation of the above-mentioned dosage form, generally known methods are applied, for example, various excipients, lubricants, binders, disintegrating agents, suspending agents, isotonic agents, emulsifying agents, absorbing agents. You may contain a promoter etc.

The carrier used in the pharmaceutical composition includes, for example, water, distilled water for injection, physiological saline, glucose, fructose, sucrose, mannitol, lactose, starch, corn starch, cellulose, methyl cellulose, carboxymethyl cellulose, Examples include hydroxypropyl cellulose, alginic acid, talc, sodium citrate, calcium carbonate, calcium hydrogen phosphate, magnesium stearate, urea, silicone resin, sorbitan fatty acid ester, glycerin fatty acid ester, etc. To be selected.

The dose of radicicol is determined according to the desired therapeutic effect, administration method, treatment period, patient's age, body weight, etc., but oral or parenteral administration methods (eg injection, infusion, suppository) It is usually 0.01 to 2 mg / kg per day for adults by rectal administration or skin patching. Next, embodiments of the present invention will be described with reference to examples.

[0018]

【Example】

Example 1 Tablets Radicicol 100 g, lactose 40 g, corn starch 18 g and carboxymethylcellulose calcium 10
g was mixed, 10% hydroxypropyl cellulose aqueous solution was added, and the mixture was kneaded by a conventional method. This kneading liquid is granulated with an extrusion granulator equipped with a 1.0 mm basket, and magnesium stearate is added to the granules to form granules for tableting.
Tableting was performed by a conventional method to give a tablet of 8 mm diameter containing 100 mg of radicicol in one preparation (170 mg).

Example 2 Capsules A mixture of 50 g of radicicol, 80 g of lactose and 38 g of potato starch was mixed with a 10% aqueous solution of hydroxypropylcellulose and kneaded, and granulated in the same manner as in Example 1 below. Magnesium stearate was added and filled into hard capsules with a capsule filling machine, and a capsule containing 50 mg of radicicol in 1 capsule (170 mg) was obtained by a conventional method.

Example 3 Soft capsule agent By dissolving 10 g of radicicol in 100 g of soybean oil and injecting the obtained solution into a capsule by a conventional method,
Soft capsules containing 10 mg radicicol per capsule were prepared.

[0021]

INDUSTRIAL APPLICABILITY The present invention provides an excellent immunosuppressive agent.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Soichiro Sato 564-5 Nakachikari, Nagaizumi-cho, Sunto-gun, Shizuoka

Claims (1)

[Claims] 1. The formula: An immunosuppressive agent containing radicicol represented by