JPS638085B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a novel anticancer agent. That is, the present invention is based on the following formula () (However, in the formula, R represents a formyl lower alkyl group, an N-lower alkylformylamino group, or a hydroxy lower alkyl group.) An anticancer agent containing a compound represented by the following as an active ingredient is provided. The present invention relates to a novel anticancer agent.
Conventionally, cancer chemotherapy agents include alkylating agents (nitrozene mustards, ethyleneimines, sulfonic acid esters), antimetabolites (folate antagonists, purine antagonists, pyrimidine antagonists), and plant fission toxins (colcemid). , vinblastine, etc.), antibiotics (sarcomycin, carcinophylline, mitomycin, etc.), hormones (adrenal steroids,
(male hormones, female hormones) and porphyrin complex salts (marphyrin, COPP) are used, but the nucleic acid inhibitory effects of anticancer substances are generally highly toxic and serious because they act not only on cancer cells but also on normal cells. At present, it is difficult to achieve sufficient effects by using large amounts of drugs such as chemotherapeutic agents for infectious diseases because they exhibit side effects. The present inventor previously developed a new and useful anticancer agent containing benzaldehyde as an active ingredient (Japanese Patent Application Laid-open No.
(Refer to Publication No. 108027), the action of this active ingredient does not directly attack cancer cells, but is thought to produce therapeutic effects through a mechanism of action that is different from that of conventionally thought chemotherapeutic agents. As a result of intensive research in search of anti-cancer active substances, we discovered that each aromatic compound of formula () has anti-cancer activity.
The anticancer agent of the present invention has now been completed based on the new knowledge that these substances can exert a remarkable effect on cancer treatment. Most conventional cancer chemotherapeutic agents are so-called cytotoxic substances, which are more sensitive to transplanted cancers such as Ehrlichi tumors than to cancerous cells caused by the SV ₄₀ oncogenic virus. The active ingredient exhibits high sensitivity by acting on cells that have become cancerous due to the SV ₄₀ oncogenic virus.
It is thought to be based on a specific anticancer effect that is different from the mechanism of action of conventional cancer chemotherapeutic agents, and has excellent characteristics as an anticancer agent. Table 1 shows examples of the aromatic compounds of formula () having anticancer activity used in the present invention.

[Table] References, physical property values, and structural formulas for the above compounds (1) to (4) (hereinafter, the above compounds are indicated by compound numbers) are shown in Table 2.

【table】

[Table] The anticancer agent of the present invention can be administered either orally or parenterally.
Administered as hard capsules, tablets, granules, fine granules, or powders; when administered parenterally, injections,
It can be administered in a dosage form such as a suppository so as to maintain sustained mucosal absorption in the form of a drop, a solid or a viscous liquid suspension. The proportion of the active ingredient in the anticancer drug composition of the present invention may vary depending on the dosage form, but usually, when administered orally or by mucosal absorption, approximately 0.3 to 15.0% by weight is appropriate; When it is, approximately 0.01 ~
10% by weight is suitable. In addition, when formulating the active ingredient of the present invention, the above-mentioned compound can be made into a subcutaneous or intravenous injection preparation in the form of an aqueous suspension, an oily preparation, etc., as well as a capsule, It can be formulated into tablets, fine granules, etc. and administered orally; however, among the above compounds, those with an aldehyde group are unstable stimulants and therefore should not be used in the formulation of compositions. It is required to prevent its deterioration and eliminate its irritating properties. These compounds include the above-mentioned compounds (1) to (3), and their autooxidation must be prevented and their stimulatory properties must be improved. For this method, for example, it is appropriate to use an inclusion compound that utilizes the inclusion ability of choleic acid or cyclodextrin, and formulation using microcapsules is also effective. In addition, in order to impart stability and acid resistance that can withstand long-term storage to the above-mentioned clathrate compounds and to fully maintain their medicinal efficacy, it is possible to formulate a formulation with a pharmaceutically acceptable coating. A stable anticancer drug composition can be obtained. The surfactants, excipients, lubricants, adjuvants, pharmaceutically acceptable film-forming substances, etc. used in formulating the active ingredient of the present invention 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, light silicic anhydride, magnesium aluminate, magnesium aluminate metasilicate, 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, fall seed oil, calcium lactate, safflower oil, soybean phospholipid, and the like. Film-forming substances include cellulose acetate phthalate (CAP) as a carbohydrate derivative such as cellulose and sugars, methyl acrylate and methacrylate as polyvinyl derivatives such as acrylic acid copolymers and dibasic acid monoesters. Examples include 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 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. The dosage depends on the desired therapeutic effect and duration of treatment, but for adults it is usually 0.5 to 5000 mg of the above compound per day, and for children it is usually 0.5 to 5000 mg per day.
It is 0.5-3000 mg. Next, the anticancer activity testing method for confirming the anticancer activity of the above compound will be described. The test cells were W2K-11 cells obtained by turning C3H mouse kidney cells into cancer with the SV ₄₀ oncogenic virus, and were cultured by the following method. (1) Preparation of growth culture solution Dissolve 9.4 g of Eagle's MEM medium in 900 ml of distilled water, autoclave at 120°C for 15 minutes, cool, add 100 ml of calf serum and 10% carbon dioxide that was separately autoclaved at 115°C for 15 minutes. Add 3-5 ml of sodium hydrogen solution
Correct to PH7.1~7.2. Immediately before use, add 10 ml of Millipore filtered L-glutamine (2.92 g/100 ml) solution. In addition, to preserve the test cells, the final concentration is 10.
% dimethyl sulfoxide. (2) Preparation of transplanted cells The test cells stored in a Jeep freezer (-80℃) were lysed at room temperature, centrifuged at 670 x g for 5 minutes, the supernatant was discarded, and the precipitated cells were suspended in 50 ml of growth medium. After it becomes cloudy, transfer it to a roux flask,
When cultured at 37°C, the cells begin to proliferate while adhering to the bottom of the flask, and fully proliferate in 3 to 4 days. Decant the culture solution, then add 0.2% trypsin solution [Eagle MEM medium (Nissui Pharmaceutical Co., Ltd.)
After dissolving 4.7 g of the solution, 0.6 g of sodium bicarbonate, and 1 g of trypsin in 500 ml of distilled water and filtering it through a Millipore filter, add 10 ml of the solution, treat with trypsin for 2 to 3 minutes at room temperature, and then decant the trypsin solution. Furthermore, add 50 ml of fresh growth medium and wash off adhering cells using a Komagome pipette to obtain a cell suspension. A portion is subcultured using Roux flasks. (3) Cell culture and administration of test compound Dispense 1.8 ml of the above cell suspension into a disposable chamber (diameter 35 mm) and incubate at 37°C in a carbon dioxide incubator (5% CO ₂ , 95% air).
Incubate for 24 hours. At this point, 0.2 ml of the test compound solution is administered to continue the culture. The state of cell proliferation is observed every day using an inverted microscope, and the number of surviving cells is counted 48 hours after administration. The test compound is dissolved in distilled water or ethanol (final concentration 2%) and then filtered through a Millipore filter. (4) How to count cells After administering the test compound, decant the 48-hour-old shell, discard the supernatant (culture medium), and treat the cells attached to the bottom of the shell with 1.0 ml of the above 0.2% trypsin solution, resulting in single cells. . Decant this to remove the trypsin solution, make a cell suspension with physiological saline containing 10 mmol of phosphate buffer (PH7.0), place 1 to 2 drops of this on a hemocytometer, and put it on a cover glass. Count the number of cells under a microscope. The inhibition rate of test cell proliferation was determined by the following formula. Inhibition rate (%) = (1 - (number of cells in the test compound-administered test compound)/(cell number in the test compound-unadministered test test sample)) x 100 Next, regarding the toxicity of the active ingredient compound of the present invention, Because of its low molecular structure, the compound is quickly excreted outside the body and does not cause any side effects.
All of the LD ₅₀ values are lower in toxicity than anticancer substances. For example, the LD ₅₀ (mg/Kg) of the above compounds (1), (2), and (3) when administered orally to mice is ,Each
3000, over 8000 and 1500. The present invention will be specifically explained below using formulation examples and test examples. Formulation Example 1 (Injection/Drop) Add 5 g of powdered glucose to contain 500 mg of the above compound (1) and aseptically dispense into vials.
Seal tightly, fill with inert gas such as nitrogen or helium, and store in a cool, dark place. Before use, add 500 ml of 0.85% physiological saline to make an intravenous injection,
Administer 10 to 500 ml per day by intravenous injection or drip depending on symptoms. Formulation Example 2 (Injection/Drop) An intravenous injection for mild symptoms was prepared in the same manner as Formulation Example 1, except that 50 mg of the above compound (3) was used.
Administer 10 to 500 ml per day by intravenous injection or drip depending on symptoms. Formulation Example 3 (Injection, Capsule) 30 mg of the above compound (2) was dissolved in 1 g of refined sesame oil and 100 mg of aluminum stearate gel, sealed, sealed with an inert gas such as nitrogen or helium, and stored in a cool, dark place. , as a preparation for subcutaneous injection. Administer 1 to 10 ml subcutaneously once a day depending on the symptoms. In addition, the above preparation is dispensed into capsules in 0.5 ml portions to prepare oral capsules, and 1 to 10 capsules are orally administered per day depending on the symptoms. Formulation example 4 (enteric-coated tablet) β-cyclodextrin (manufactured by Nihon Shokuhin Kako Co., Ltd.)
Add 15 g of the above compound (3) to 3,000 ml of a saturated aqueous solution of can get. Enteric-coated tablets (1) for adults and 1000 for children (2) were manufactured with the following ingredient composition.

[A]

Main ingredient (clathrate compound of compound (1) above) 100 (g) Lactose 737 Hydroxypropyl cellulose 3 [B] Cellulose acetate phthalate 80 (g) Hydroxypropyl methyl cellulose phthalate
80 After thoroughly mixing the ingredients of [A], they were formed into granules according to a conventional method, and the granules were thoroughly dried and sieved to produce granules suitable for bottles, heat-seal packaging, etc. Next, the granules are coated with the dissolved base material [B] while floating and flowing to form enteric-coated granules. When this granule was subjected to a disintegration test using a Japanese Pharmacopoeia disintegration tester, it did not disintegrate even when shaken in artificial gastric juice of pH 1.2 for one hour. In artificial intestinal fluid with a pH of 7.5, it disintegrated in 5 minutes. Formulation Example 6 (Enteric-coated capsules) 1000 enteric-coated capsules were manufactured using the following ingredients.

[Table] Cellulose phthalate Enteric-coated granules suitable for capsules were prepared using the above ingredients in the same manner as described in Formulation Example 5, and the composition was filled into capsules to obtain enteric-coated capsules. When this capsule was subjected to a disintegration test using a Japanese Pharmacopoeia disintegration tester, no disintegration or dissolution was observed even after shaking in artificial gastric fluid at pH 1.2 for 1 hour, and no disintegration or dissolution was observed in artificial intestinal fluid at pH 7.5 in 5 minutes. Collapsed or the entire amount eluted. Test Example 1 Using the above compounds (1) to (4) as aromatic compounds,
When the growth inhibition rate (%) of cancer cells W2K-11 in C3H mice that had become cancerous due to the SV ₄₀ oncogenic virus was calculated using the above test method, the results shown in Table 3 were obtained. At the same time, the LD ₅₀ (mg/Kg) of each compound was also written.

[Consideration]

As is clear from the results of the above test examples, the above test compound was proven to have excellent anticancer activity. In addition, the above test compound has an extremely low activity against cancerous cells caused by the SV ₄₀ oncogenic virus, whereas most of the conventional cancer chemotherapeutic agents have an extremely low activity against cancerous cells caused by the SV 40 oncogenic virus. It should be noted that it is extremely characteristic that both compounds exhibit extremely low toxicity. In other words, many of the conventional cytotoxic anticancer drugs show remarkable activity against experimental tumors in animals, that is, transplanted cancers, but clinically many problems remain, and there are very few effective cases. This is the actual situation, and this is thought to be based on the fundamental difference between transplanted cancer and primary cancer, and can be said to be an artificial primary cancer.
The above-mentioned compound, which is active against SV ₄₀ virus-induced cancer and has low toxicity, is recognized to have very specific anticancer activity.

Claims (1)

[Claims] Linear formula () (However, in the formula, R represents a formyl lower alkyl group, an N-lower alkylformylamino group, or a hydroxy lower alkyl group.) An anticancer agent containing a compound represented by the following 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.