JPH0330570B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a novel anticancer agent characterized by containing 2,6-di-tert-butyl-p-cresol as an active ingredient. Conventionally, alkylating agents (nitrodiene mustards, ethyleneimines,
sulfonic acid esters), antimetabolites (folate antagonists, purine antagonists, pyrimidine antagonists), plant fission toxins (colcemid, vinblastine, etc.),
Antibiotics (sarcomycin, carcinophilin,
mitomycin, etc.), hormones (steroids, male hormones, female hormones), and porphyrin complex salts (marphyrin, copp), etc. are used. However, most of them are cytotoxic substances and exhibit serious side effects, so the development of anticancer agents with low toxicity and excellent anticancer activity is strongly desired. Therefore, in view of the above-mentioned purpose, the present inventors searched for a substance with low toxicity and anticancer activity, and as a result of intensive research, 2,6-di-tert-butyl-p-cresol was found to be effective against tumor cells in animals. Obtaining new findings that the substance has differentiation-inducing activity against cancer, has extremely low toxicity, and has excellent anticancer activity,
The anticancer agent of the present invention has been completed. The active ingredient of the anticancer agent of the present invention can serve as an excellent cancer chemotherapeutic agent for humans, livestock, warm-blooded animals such as dogs and cats. 2,6-di-2,6-di-carcinogen having anticancer activity used in the present invention
Tert-butyl-p-cresol will be explained below. 2,6-di-tert-butyl-p-cresol is commonly called "BHT" and can be easily obtained by heating p-cresol and isobutylene in the presence of hydrogen fluoride, anhydrous aluminum chloride, etc. It is a substance and its physical properties are as follows. (1) mp: 69-70℃, (white crystals with a slight phenolic odor) (2) bp: 265℃ (3) Solubility: Insoluble in water, soluble in ethanol, petroleum, cottonseed oil, and peanut oil. Moreover, the toxicity is LD ₅₀ 1.3g/Kg (mouse oral). The anticancer agent of the present invention can be administered either orally or parenterally.
Administered as hard capsules or tablets, granules, fine granules, or powders, and for parenteral administration, subcutaneous or intravenous injections such as aqueous suspensions and oil-based preparations, infusions, and solid or suspended viscosity. It can be administered in the form of a suppository so that sustained mucosal absorption can be maintained in the form of a thick liquid. The formulation of the active ingredient of the present invention includes surfactants, excipients, lubricants, adjuvants, and, if necessary, pharmaceutically acceptable film-forming substances and coating aids to form enteric-coated formulations. Specific examples thereof are as follows. In order to improve disintegration and elution of the composition of the present invention, one or more surfactants such as alcohols, esters, polyethylene glycol derivatives, sorbitan fatty acid esters, sulfated fatty alcohols, etc. are added. Can be added. In addition, excipients such as sucrose, lactose, starch, crystalline cellulose, mannite, soft silicic anhydride, magnesium aluminate, magnesium metasilicate aluminate, synthetic aluminum silicate, calcium carbonate, sodium hydrogen carbonate, calcium hydrogen phosphate, carboxylic Methylcellulose calcium and the like can be added alone or in combination of two or more. As a lubricant, for example, one or more types of magnesium stearate, talc, hydrogenated oil, etc. can be added, and as a flavoring agent and a flavoring agent,
Sweeteners such as salt, saccharin, sugar, mannitrite, orange oil licorice extract, citric acid, glucose, menthol, eucalyptus oil, and malic acid, fragrances, colorants, preservatives, and the like may be included. Adjuvants such as suspending agents and wetting agents may include, for example, coconut oil, olive oil, sesame oil, peanut oil, calcium lactate, safflower oil, soybean phospholipid, and the like. Film-forming substances include cellulose, cellulose acetate phthalate (CAP) as a carbohydrate derivative such as sugars, acrylic acid copolymers, and polyvinyl derivatives such as dibasic acid monoesters such as methyl acrylate and methacrylic acid copolymers. Examples include polymers 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 additives to prevent chemicals from adhering to each other during coating operations can be added to the film-forming agent. properties and make coating operations easier. In addition,
It is also possible to form a dosage form in which the active ingredient is microencapsulated using a film-forming substance and then mixed with excipients and the like. In particular, the blending ratio in typical dosage forms is as follows.

[Table] Particularly preferred excipients are lactose, crystalline cellulose, and carboxymethylcellulose calcium. In addition, the dosage is sufficient to effectively treat the target tumor, and depends on the symptoms of the tumor, route of administration, dosage form, etc., but in general, in the case of oral administration, the amount per day for adults is The range is about 0.01 to 100 mg/Kg body weight (0.01 to 60 mg/Kg body weight for dwarfs), and the upper limit is preferably about 50 mg/Kg body weight, more preferably about 10 mg/Kg body weight, and it is suitable for parenteral administration. In this case, the upper limit is approximately 10 mg/Kg body weight, preferably 5 mg/Kg body weight, and more preferably 2 mg/Kg body weight. Next, we will discuss anticancer tests that confirmed the anticancer activity of BHT. [1] Friend leukemia cells (mouse erythroid
leukemia cell, B8 cells) B8 cells were inoculated at 2.5 x 10 ⁴ cells/ml into GIBCO HAM F-12 medium supplemented with 15% fetal bovine serum and 60 mg/kanamycin. Then, add a predetermined amount of the test compound (final volume: 5 ml). After culturing for 7 days at 37°C in 7.5% _CO2 , the cells were stained using Orkin's benzidine staining method, and the number of stained cells, that is, the number of cells that began to produce hemoglobin due to differentiation into red blood cells, was determined. Measure and determine the differentiation induction rate. Differentiation induction rate (%) = number of stained cells/total number of cells x 10
0 [2] Mouse teratoma cells (teratocarcinoma cells)
Teratoma cells were used one month after being transplanted from the peritoneal cavity of a mouse to the peritoneal cavity. Teratoma cells exist in the peritoneal cavity as a cell mass called an embryoid body, which resembles an early embryo.
They were used without trypsin treatment. The embryoid bodies obtained by natural settling in the collected ascites were washed three times with Dulbecault's modified medium or Hank's solution, then inoculated into a medium containing 10% fetal bovine serum, and a predetermined amount of the test compound was added. ℃
The cells are saturated with water vapor containing 7.5-8% CO ₂ and cultured in air for one week. Centrifugation (2000r.p.
After washing the embryoid bodies obtained using 0.86% NaCl solution, naphthol AS-MX phosphate and diazo reagent (Fast Violet B Salt) were added and left at room temperature for 1 hour. This is centrifuged (2000 rpm, 10 minutes) to separate the embryoid bodies, ethanol is added, and the mixture is left at room temperature for 1 hour. (Undifferentiated cells are colored red). this,
Measure the absorption at 535 nm to determine alkaline phosphatase activity (degree of differentiation induction). When 5mM hexamethylene bisacetamide (HMBA) is added (shows no alkaline phosphatase activity at all), it is marked as "++", and when HMBA is not added (shows extremely strong alkaline phosphatase activity), it is marked as "-". -'' and the degree of differentiation induction was shown in the following stages. ++: Shows no alkaline phosphatase activity. +: Almost no alkaline phosphatase activity. ±: Slightly shows alkaline phosphatase activity. -: Strongly shows alkaline phosphatase activity. --: Shows extremely strong alkaline phosphatase activity. In addition, in the test examples described below, it had a differentiation-inducing effect and showed anticancer activity. The present invention will be specifically explained below using formulation examples and test examples. Formulation example 1 (injection/infusion) 5g of powdered glucose to contain 10mg of BHT
Aseptically dispense into vials, seal tightly, fill with inert gas such as nitrogen or helium, and store in a cool, dark place. Dissolve in ethanol before use.
Add 100 ml of 0.85% physiological saline to make an intravenous injection, and administer 10 to 100 ml per day by intravenous injection or drip depending on the symptoms. Formulation Example 2 (Injection/Drip) Using 2 mg of BHT, prepare an intravenous injection for mild symptoms using the same method as Formulation Example 1, and administer 10 to 100 ml per day by intravenous injection or drip depending on the symptoms. Formulation Example 3 (Enteric-coated capsules) Take 5g of BHT, 2.46g of lactose, and 0.04g of hydroxypropyl cellulose, mix well, and then
The product is formed into granules according to a conventional method, thoroughly dried and sieved to produce granules suitable for bottles, heat-seal packaging, etc. Next, cellulose acetate phthalate
0.5 g and 0.5 g of hydroxypropyl methyl cellulose phthalate are dissolved to form a coated base material, and the base material is coated while the granules are suspended and flowed to form enteric-coated granules. This composition is filled into capsules to produce 100 enteric-coated capsule preparations. Test Example Using BHT, the differentiation induction rate of Friend leukemia cells and the degree of differentiation induction of mouse teratoma cells were investigated using the above test methods [1] and [2], and the results are shown in Tables 1 and 2, respectively. The following results were obtained.

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[Table] As is clear from the results of the above test examples, BHT
It has been demonstrated that it exhibits excellent anticancer activity with little toxicity, as it shows an effect on inducing cancer cells to differentiate into normal cells.

Claims (1)

[Scope of Claims] 1. An anticancer agent characterized by containing 2,6-di-tert-butyl-p-cresol as an active ingredient. 2. The anticancer agent according to claim 1 in a parenteral administration form. 3. The anticancer agent according to claim 1 in an oral administration form.