JPH0617304B2 - Anti-cancer drug - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention provides the following compounds which show differentiation-inducing activity on cancer cells: Or The present invention relates to an anticancer drug containing as an active ingredient.

Conventionally, as cancer chemotherapeutic agents, alkylating agents (nitrodien mustards, ethyleneimines, sulfonates), antimetabolites (folate antagonists, purine antagonists,
Pyrimidine antagonist), phytofission venom (corsemide, vinblastine, etc.), antibiotics (sarcomycin, carcinophylline, mitomycin, etc.), hormones (adrenal steroids, male hormones, female hormones), and porphyrin complex acid (murphyrin, copp) Etc. are used.
However, most of them are cytotoxic substances,
Since it has serious side effects, it is strongly desired to develop an antitumor agent having low toxicity and excellent antitumor activity.

Therefore, in view of the above-mentioned gist, the present inventors searched for a substance having low toxicity and antitumor activity, and as a result of earnest research, the flavonoid or isoflavonoid compound has a differentiation-inducing activity on animal tumor cells. The present inventors have newly found that they have, and obtained new knowledge that the substance has extremely low toxicity and excellent antitumor activity, and completed the antitumor agent of the present invention. The active ingredient of the antitumor agent of the present invention can be an excellent cancer chemotherapeutic agent for warm-blooded animals such as humans, livestock, dogs and animals.

The flavonoids or isoflavonoids which are the active ingredients of the present invention are the following compounds.

(1) Flavone (2) Fisetin: 3,3 ', 4', 7-tetr
ahydroxyflavone (3) Naringenin: 4 ', 5,7-trihy
droxyflavanone (4) Flavanone (Flavanone) As shown in Table 1, all of these compounds are known compounds, and their structural formulas and physical properties are as follows (hereinafter, indicated by the above compound numbers). .).

The carcinostatic agent of the present invention can be used either orally or parenterally, and when orally administered, it is administered as soft / hard capsules or tablets, granules, fine granules, powders, and parenterally. Can be administered in subcutaneous or intravenous injections such as aqueous suspensions, oily preparations, infusions and solid or suspension viscous liquids in the form of suppositories so that sustained mucosal absorption can be maintained. .

The active ingredient of the present invention is formulated by a surfactant, an excipient, a lubricant, an adjuvant, and a pharmaceutically acceptable film-forming substance or a coating aid to form an enteric-coated preparation as required. Can be appropriately performed by using, for example,
It is as follows.

In order to ensure good disintegration and dissolution of the composition of the present invention,
Surfactants such as alcohols, esters, polyethylene glycol derivatives, fatty acid esters of sorbitan, and sulfated fatty alcohols can be added alone or in combination of two or more.

Further, as an excipient, for example, sucrose, lactose, starch, crystalline cellulose, mannitol, light anhydrous silicic acid, magnesium aluminate, magnesium metasilicate aluminate, synthetic aluminum silicate, calcium carbonate, sodium hydrogen carbonate, calcium hydrogen phosphate, carboxy One kind or two or more kinds such as methylcellulose calcium can be added in combination.

As the lubricant, for example, magnesium stearate, talc, hardened oil and the like can be added alone or in combination of two or more, and as a flavoring agent and a flavoring agent, salt, satukaline,
Sweeteners such as sugar, mannitol, orange oil licorice extract, citric acid, glucose, menthol, eucalyptus oil, malic acid, etc., flavors, colorants, preservatives and the like may be contained.

Examples of adjuvants such as suspending agents and wetting agents include coconut oil, olive oil, sesame oil, peanut oil, calcium lactate,
Safflower oil, soybean phospholipid, etc. can be contained.

Further, as a film-forming substance, cellulose, cellulose acetate phthalate (CAP) as a carbohydrate derivative such as saccharide,
Examples of polyvinyl derivatives such as acrylic acid-based copolymers and dibasic acid monoesters include methyl acrylate / methacrylic acid copolymers and methyl methacrylate / methacrylic acid copolymers.

In addition, when coating the above film-forming substance, a coating-forming agent which is usually used, for example, a plasticizer and various additives for preventing mutual adhesion of chemicals during the coating operation are added to form a film-forming agent. You can improve the properties of the and make the coating operation easier. The active ingredient may be microencapsulated using a film-forming substance and then mixed with an excipient or the like to give a dosage form.

In particular, the compounding ratio in a typical formulation is as follows.

Particularly preferred excipients are lactose, crystalline cellulose and carboxymethyl cellulose calcium.

In addition, the dose is an amount sufficient to effectively treat the target tumor and depends on the tumor symptom, the administration route, the dosage form, etc. About 0.01 to 100 mg / kg body weight (0.01 to 60 for dwarfs)
mg / kg body weight), the upper limit is preferably about 50 mg
/ Kg body weight, more preferably about 10 mg / kg body weight. In the case of parenteral administration, the upper limit is about 10 mg / kg body weight, preferably 5 mg to kg body weight, more preferably 2 mg / kg body weight.
mg / kg body weight is appropriate.

Next, a carcinostatic test confirming the carcinostatic activity of the compound of the present invention will be described.

[1] Friend leukemia cell (mouse erythroid leukemia
cell, B8 cells) GIBCO HAM F-12 medium supplemented with 15% fetal bovine serum and 60 mg / l kanamycin was inoculated with B8 cells at 25 × 10 ⁴ cells / ml. Then, a predetermined amount of the test compound is added thereto (final volume 5 ml).

After culturing in 7.5% CO ₂ at 37 ° C for 7 days,
The number of cells stained by the benzidine staining method of n), that is, the number of cells adapted to produce hemoglobin due to differentiation into erythrocytes is measured, and the differentiation induction rate is obtained.

[2] Mouse myeloid leukem
Test for ia cell, M1) GIBCO Eagle MEM medium supplemented with 10% horse serum and 60 mg / l kanamycin 5.0 × 10
M1 cells are inoculated to obtain ⁴ cells / ml, and a predetermined amount of the test compound is added thereto (final volume 5 ml).

After culturing in 7.5% CO ₂ at 37 ° C. for 7 days, lysozyme activity induced by differentiation into phagocytes or granulocytes is examined. In addition, 1 unit of lysozyme activity (unit),
Micrococcus lycus
odeikticus) A suspension of microbial cells is used as a substrate, and lysozyme is allowed to act on it, and the amount of lysozyme is measured at pH 6.24 and a temperature of 25 ° C. to reduce the absorbance at a wavelength of 450 mμ by 0.001 per minute.

[3] Test for mouse teratocarcinoma: One month after teratoma cells were transplanted from the abdominal cavity of the mouse to the abdominal cavity, the cells were used. Teratoma cells exist in the abdominal cavity as a cell mass called an embroid body similar to the early embryo, and they were used without trypsin treatment. The embryoid body obtained by spontaneously submerging in the collected ascites was washed 3 times with a modified Darbeco's medium or Hanks' solution, and then inoculated into a medium containing 10% fetal bovine serum, and a predetermined amount of the test compound was added to the medium. CO ₂ 7.5-8 at ℃
% Of water vapor is saturated and incubated in air for 1 week.
The embryoid body obtained by centrifugation (2000 rpm 10 minutes) was
After washing with 86% NaCl solution, naphthol AS-MX phosphate and diazo reagent (Fast Violet B Salt) are added and left at room temperature for 1 hour. This is centrifuged (2000r.pm,
After 10 minutes), the embryoid bodies are separated, ethanol is added, and the mixture is left at room temperature for 1 hour.

(Undifferentiated cells are colored red).

The absorption at 535 nm is measured to determine the alkaline phosphatase activity (degree of differentiation induction).

The case where 5 mM of hexamethylene bisacetamide (HMBA) was added (has no alkaline phosphatase activity at all) was designated as "++", and the case where HMBA was not added (has extremely strong alkaline phosphatase activity). --"age,
The degree of differentiation induction was shown in the next step.

++: shows no alkaline phosphatase activity at all.

+: Almost no alkaline phosphatase activity is shown.

+: Shows some alkaline phosphatase activity.

-: Strongly shows alkaline phosphatase activity.

-: Extremely strong alkaline phosphatase activity.

In addition, in the test example described later, it has anti-tumor activity by having a differentiation inducing action.

Hereinafter, the present invention will be specifically described with reference to formulation examples and test examples.

Formulation example (enteric coated capsule) Compound (1) 5 g, lactose 2.46
g and hydroxypropylcellulose (0.04 g) are mixed well, and then the mixture is shaped into granules by a conventional method, dried well and sieved to produce granules suitable for bottles, heat-sealed packaging and the like. Next, 0.5 g of cellulose acetate phthalate and 0.5 g of hydroxypropylmethylcellulose phthalate are dissolved to form a coated base material, and the above granules are allowed to float and flow, and this base material is coated to form an enteric coated granule. This composition is filled into capsules to produce 100 enteric-coated capsule preparations.

Test Example From the test methods [1], [2] and [3] using the compounds of Table 1, the induction rate of differentiation of friend leukemia cells, lysozyme activity by induction of differentiation of mouse myeloid leukemia cells and mouse teratoma cells When the degree of differentiation induction was examined, the results shown in Table 2, Table 3 and Table 4 were obtained.

As is clear from the results of the above Test Examples, the various flavonoids and isoflavonoids of the present invention show an effect of inducing differentiation into normal cells against cancer cells, and thus exhibit excellent antitumor activity with low toxicity. Proven.

Front Page Continuation (56) References Japanese Patent Publication No. 53-23832 (JP, B1) Japanese Patent Publication No. 53-23831 (JP, B1) Journal of National Products vol. 42, No. 1, P. 85-P. 91, (1979)

Claims (1)

[Claims] 1. The following compound showing a differentiation-inducing activity against cancer cells: Or An anticancer agent containing as an active ingredient.