JPS60178815A - Carcinostatic agent - Google Patents

Abstract

PURPOSE:A carcinostatic agent having extremely low toxicity and improved carcinostatic activity, comprising a specific compound such as chalcone, flavanone, etc. CONSTITUTION:A carcinostatic agent containing a compound selected from chalcone, dihydrochalcone, flavanone, isoflavanone, rotenoid, isoflavan, isoflavene, pterocarpan, coumestan, 3-arylcoumarin derivative, and dimethylmelannein as an active ingredient. The compound was found newly to have differentiation and introduction activity against animal tumor cells. An amount of the compound in a pharmaceutical preparation is preferably 0.3-15wt%, a daily dose is 0.01- 100mg per kg weight of adult in case of oral administration, the upper limit is 50mg, especially 10mg, and the upper limit of parenteral administration is 10mg, especially 2mg.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention provides chalcone, dihydrochalcono
s flavanone, y (1soflavanone).

rotinoid (rotenold), infranogun (
1so-i1avan), Infrapen (1sofl
avene), zoterocalzz:y (pteroc
arpan), Kumestay (Coumest13n)
), 3-arylcoumar
ln) derivatives and dimethylmelanein (dlmeth
16 (1), which is characterized by containing a compound selected from ylmelanneln) as an active ingredient.

Conventionally, alkylating agents (nitrozoene mustards, ethyleneimines, etc.) have been used as cancer chemotherapy agents.

sulfonic acid esters), antimetabolites (folate antagonists,
purine antagonists, pyrimidine antagonists), plant fission toxins (
colcemid, vinblastine, etc.), antibiotics (sarcomycin, carcinophylline, mitomycin, etc.), hormones (adrenal steroids, male hormones, female hormones), and volufiline lead acid (marfilin, copp)
etc. are used. However, most of them are
Since it is a cytotoxic substance and exhibits serious side effects, there is a strong desire to develop an anticancer agent with low toxicity and excellent anticancer activity.

Therefore, in view of the above-mentioned purpose, the present inventors have developed a
511 As a result of extensive research and exploration of substances that have cancer activity, it was newly discovered that the compound has differentiation-inducing activity that is beneficial to animal tumor cells, and that the substance has extremely low toxicity and is an excellent anticancer agent. We have obtained new knowledge regarding the presence of active gold, and have completed the anticancer agent of the present invention. The active ingredient of the anticancer agent of the present invention is:

It can be an excellent cancer chemotherapeutic agent for humans, livestock, dogs, cats, and other animals.

Examples of compounds that are active ingredients of the present invention include the following compounds.

As a chalcone compound.

(a) llqulrltige
nin ) (b) isori kiri teizo x nin (lso
(c) As a flavanone compound, (d) Licoflapanone (IIcOi1avanone)
) As an isoflavanone compound, (e) 3,4,6.7-titramethoxyflavanone (3
,4,6,7-tetramatoxyflavan
one ) (1> Daizui 7 (daldzln)
(g) Licoinflavon A (1lcolsofla
bone A) (h) co-lsoflavons B tri
acetate) as a rotinoid compound, (+) rotenone as an uncompounded isoflavan, medium bestitol/l/ (vestltol) (k
) Licolicynon (1lcorlcdin) (1
) Glabridin 7 (glabridin) As an isoflaven compound, (m) 5yn-isofujihe:y (5yn-1sof
lavsne ) (n) glabre
ne )7' Theocarno9one-based compound, (0)
Demethylhomobutyroruby (demethylh)
omopterocarpin) (p)? - As a coumestane compound.

(q) Glycyrol 3-allylcoumarin compound, (r) syn-3-allylcoumarin (5yn-3
-aryl-coumarin) (s) glycyrin Other compounds include (t) 7y-homoiso7ugin (syn-homo
iso-flavone) (u) dimethylmelanein (dlmethylmel
anneln) All of these compounds are known compounds, and their structural formulas are as shown in Table 1.

Table 1 Compound Number Structural Formula () () % Formula % 0 The anticancer agent of the present invention can be administered either orally or parenterally, and when administered orally, Administered as soft or hard tablets, tablets, granules, fine granules, or powders (for parenteral administration, aqueous suspensions,
It can be administered in the form of crude drugs, such as subcutaneous or intravenous injections such as oil-based preparations, infusions, and solid or suspended viscous liquids to maintain sustained mucosal absorption.

The formulation of the active ingredient of the present invention includes surfactants, excipients,
This can be carried out as appropriate using lubricants, adjuvants, and, if necessary, pharmaceutically acceptable film-forming substances and coating aids to prepare enteric-coated preparations. Specific examples include the following: In order to improve the disintegration and dissolution of the composition of the present invention, surfactants such as alcohol,
Esters, SiJ? Liethylene glycol derivatives, fatty acid esters of sorbitan, sulfated fatty alcohols, etc. can be used to add two or more additives.Additionally, excipients such as sja, lactose, denone, crystalline cellulose, mannitol, etc. One or more of light silicic anhydride, magnesium aluminate, meta-magnesium aluminate, synthetic aluminum silicate, calcium carbonate, sodium hydrogen carbonate, calcium hydrogen phosphate, calcium carboxymethyl cellulose, etc. or 24 JU or more may be added in some cases. can.

As a lubricant, for example, one or more types of stearin, magnesium stearin, talc, hydrogenated oil, etc. can be added, and as a flavoring and flavoring agent, salt, saccharin, sugar, mannitol, orange oil licorice extract, etc. can be added. ,citric acid,
Sweeteners such as glucose, menthol, eucalyptus oil, and malic acid, flavoring agents, coloring agents, and preservatives? 4j may be included.
Suspending agent? ! Examples of adjuvants such as agent #J include coconut oil, olive oil, sesame oil, quince oil, calcium lactate, safflower oil, soybean phospholipid, and the like.

Film-forming substances include cellulose, cellulose acetate phthalate (CAP) as carbohydrate derivatives such as sugars,
Examples of polyvinyl derivatives such as acrylic acid copolymers and dibasic acid monoesters include methyl acrylate/methacrylic acid copolymers and methyl methacrylate/methacrylic acid copolymers.

In addition, when coating the above-mentioned film-forming substance, in addition to commonly used coating aids such as plasticizers, various persimmon additives can be added to prevent adhesion of chemicals to each other during coating operations. The properties 19 of the forming agent can be improved and the coating operation can be made easier. Note that the active ingredient may be microencapsulated using a film-forming substance and then mixed with excipients and the like to form a dosage form.

In particular, the blending ratio in a typical dosage form is as shown below.

Particularly preferred Its active ingredient range: 0.1-9ON content%: 0.3-15M content: Excipients: 10-99.8 #85-99.4 #Lubricant
o~5oIo~2ol Surfactant 0~5o l O~2ol Film forming substance 0.1~50 #0.5~20 #Particularly preferred excipients are lactose, crystalline cellulose, carboxymethylcellulose calcium .

In addition, the dosage is sufficient to effectively treat the target tumor, and it depends on the symptoms of the tumor, route of administration, dosage form, etc., but in general, in the case of oral administration, it is enough to treat the target tumor once per day for adults. Approximately o, oi ~1[]Om9/Kg (L for dwarfs)
The upper limit is preferably about 5Ω/kg body weight, more preferably about 1Ω/kg body weight, and in the case of parenteral administration, the upper limit is Approximately 1 omy/+rp body weight, preferably 5-9/cm body weight, more preferably 2-/CP body weight.

Next, what about the compounds of the present invention? We will briefly describe the Sk inhibition test that confirmed lft1 cancer activity.

[1] Friend leukemia cells (mouse erythr
oidleukemia cel +, 88651
Test on GIBCO HAM F-12
25 X 10' cells were added to the medium supplemented with 15% fetal bovine serum and 60 mf//l kanamycin.
/awl! So that B8 thin Qi! ! to which a predetermined amount of the test compound is added (final volume: 5 m).

After culturing for 7 days at 57°C in 7.5% CO2, the cells were stained with Orkin's benzonor staining method to determine the number of stained 6'l cells, i.e., the number of stained hemoglobin cells produced by differentiation into red blood cells. Measure the number of cells that have become differentiated and calculate the differentiation rate.

[2] Mouse myeloid leukemia cell 11f ((mouse m
yellowleukemia cell, M1)
Test for G I 8CO excretion in single MEM medium,
5, OX 10' cell/me to 10% horse mackerel YH and 60 m/m kanamycin
[M1 thin@] was airlifted, and to this was added n [amount of test compound (final volume: 5tRt).

After culturing for 7 days at 57°C in 7.5% CO2, the lysozyme activity induced by differentiation into phagocytic cells and certain 1nFi granulocytes is examined. In addition, the 1NL position of lysozyme activity (
unit') is Micrococcus Iysodeikti.
Cus) Using a suspension of bacterial cells as a substrate, lysozyme was allowed to act, and the measurement was carried out at 25°C at 25°C.
It refers to the amount of zozyme that reduces the absorbance at wavelength μ by 0.001 per minute.

[3] Mouse teratoma 111
Test against teratoma (tocarcinoma) @: Teratoma cells were used one month after being transplanted from the peritoneal cavity of a mouse to the peritoneal cavity.

In the peritoneal cavity, teratoma cells form embryoid bodies (e
mbrold body), and these were used without treatment with trigsin or the like.

After washing the embryoid bodies obtained by natural settling in the collected ascitic fluid three times with Dulbecault's modified medium or I. Nx's solution,
It is inoculated into a medium containing 10% tallow rot, added with a predetermined amount of the test compound, saturated with water vapor containing 7.5% to 8% CO2 at 37°C, and cultured in air for one week. Centrifugation (
2000 r, p, m, 10 min) The embryos obtained were 0.
Cleaned with 86% NaCj solution, naphthol AS-MX
Phosphate and Diazo Reagents (Fast Violet
B 5alt) was added and left at room temperature for 1 hour. This was centrifuged (2000r, p, m).

After 10 minutes), the embryoid bodies are separated, ethanol is added, and the cells are left at room temperature for 1 hour.

(Undifferentiated cells are colored red).

The absorption at 5550m was measured, and 5mM of hexamethylene bisacetamide (HMBA) was added to calculate the alkaline phosphatase activity (degree of differentiation induction).
8'4 (does not show alkaline phosphatase activity at all) ir++J, and "-1" if no HMBA is added (extremely shows alkaline phosphatase activity)
The degree of differentiation induction was shown in the following steps.

++: O showing no alkaline phosphatase activity +: Almost no alkaline phosphatase activity.

- Shows some alkaline phosphatase activity.

- Strongly exhibits dual-alkaline phosphatase activity.

--: Shows very strong alkaline phosphatase activity.

In addition, in the test examples described below, it showed anticancer activity with a differentiation-inducing effect.

The present invention'f will be specifically explained below using formulation examples and test examples.

Formulation Example 1 (Injection/Drop) Add 5f of powdered glucose to contain 101 ng of compound (a), dispense into vials aseptically, seal, and fill with inert gas such as nitrogen or helium. Store in a cool, dark place. Before use, dissolve in ethanol and add 100a/l of 0.85% physiological saline to make an intravenous injection.
Administer 10 to 10 logos by intravenous injection or infusion depending on the symptoms. Formulation Example 2 (Injection/Drop) Using 2 mg of compound (g), administer intravenously for mild symptoms in the same manner as Formulation Example 1. As an intravenous injection, 10 to 100 doses per day
Administer ml by intravenous injection or infusion depending on symptoms.

Formulation Example 5 (Enteric Coated Capsules) Compound 52, milk m2.46, and hydroxyzorobyl cellulose 0.041Pe were mixed, formed into granules according to a conventional method, and dried thoroughly. Insect 1
Part 1: Manufacture granules suitable for bottles, heat-seal packaging, etc. Next, cellulose acetate phthalate 0.5F and hydroxyglobil methylcellulose phthalate 0.5F.
1. The granules are suspended and flowed and the base material f! : Coated to form enteric-coated granules. This cultivated acid product is filled into capsules to produce 100 enteric-coated capsule preparations.

Using the compounds in Test Example Table 1, the above test method [:1], (2
) and [3], respectively, Friend leukemia cells (EL
C) Differentiation induction rate of mouse myeloid leukemia cells (M L
When we investigated the lysozyme activity induced by differentiation in C) and the degree of induction of differentiation in mouse teratoma tumor 1 (TER), we found that
The results shown in Tables 2, 3 and 4 were obtained, respectively.

Table 2 Table 3 Table 4 Table 4 As is clear from the results of the above test examples, each active ingredient compound of the active ingredient of the present invention induces differentiation into 11 normal cells from 11 cancer cells. It has been proven that the drug has excellent anticancer activity with little toxicity.

Patent applicant Continuation of RIKEN page 1 ■Int, C1,4 Identification symbol Office M Procedural amendment (method) Commissioner of the Patent Office Kazuo Wakasugi 1, Indication of case Patent application No. 33756, 1982 2 Invention Name System Cancer Drug 3, Relationship with the person making the amendment Applicant name (679) RIKEN 4, Agent

Claims (3)

[Claims] (1) Chalcone, dihydrochalcone
, 7lavanone (Iava-nono), Ioflavanone (l5oflavanone), rotenold (rotenold), Infrapan (Iso-fl)
avan), infrapens (1soflavens)
), Zoterocarpan *
Coumestan, a derivative of 3-arylcoumarin (6-arylcoumarn) and dimethylmelanei (dlmethylm)
An anticancer agent characterized by containing a compound selected from the group consisting of ``elanneln'' as an active ingredient. (2) The anticancer agent according to claim 1 in a parenteral administration form. (3) The anticancer agent according to claim 441 in an oral administration form.