JPS6344725B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a novel anticancer agent.
Traditionally, cancer chemotherapy agents include alkylating agents (nitrozene mustards, ethyleneimines, sulfonic acid esters), antimetabolites (folate antagonists,
purine antagonists, pyrimidine antagonists), plant fission toxins (colcemid, vinblastine, etc.), antibiotics (sarcomycin, cartinophilin, mitomycin, etc.), hormones (adrenal steroids, male hormones, female hormones), and porphyrin complex salts (marphyrin). , COPP), etc. However, the nucleic acid inhibitory effects of anticancer substances are generally highly toxic as they act not only on cancer cells but also on normal cells.
At present, it is difficult to achieve sufficient effects by using large doses of drugs such as chemotherapeutic agents for infectious diseases, as they exhibit serious side effects. The present inventor previously developed a new and useful anticancer agent containing benzaldehyde as an active ingredient (patent no.
1157737), the action of this active ingredient does not directly attack cancer cells, but is a specific drug that is thought to produce therapeutic effects through a mechanism of action that is different from that of conventionally thought chemotherapeutic agents. They newly discovered that it has anticancer activity, and as a result of intensive research to search for substances with anticancer activity, they discovered substances with anticancer activity in aromatic aldehydes, and these substances have a remarkable effect on cancer treatment. We have now completed the anticancer agent of the present invention by obtaining new findings regarding its ability to exert its effects. 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. Aromatic aldehydes having anticancer activity used in the present invention are shown in Table 1.

[Table] References and physical property values for the above compounds 1 to 7 (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 approximately 0.3 to 15.0% by weight is appropriate when administered as a suspension or absorbed into the mucous membrane, and for parenteral administration. When administered, approximately 0.01
~10% by weight is suitable. When formulating the active ingredient of the present invention, since the above-mentioned aromatic aldehyde compound is an unstable stimulant, it is necessary to prevent its deterioration and remove its irritating properties in the formulation of the composition. Therefore, it is necessary to prevent its autoxidation and improve its stimulatory properties. 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, soft silicic anhydride, magnesium aluminate, magnesium metasilicate amine, 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, malic acid, flavoring agents, coloring agents, preservatives, etc. 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 acetate phthalate (CAP) as a carbohydrate derivative such as cellulose and sugars, acrylic acid copolymers, and methyl acrylate and methacrylate as polyvinyl derivatives such as dibasic acid monoesters. copolymer,
Examples include methyl methacrylate/methacrylic acid copolymer. 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. Although the dosage depends on the desired therapeutic effect and duration of treatment, adults usually receive 0.5 to 5000 mg of the above compound per day, and children usually receive 0.5 to 5000 mg of the compound 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% autoclaved separately at 115°C for 15 minutes.
Add 3-5ml of sodium bicarbonate solution to pH7.1~
Adjust to 7.2. Millipore filtered L-Glutamine (2.92g/100
ml) Add 10ml of solution. In addition, for the preservation of test cells, the final concentration is 10%.
of dimethyl sulfoxide. (2) Preparation of transplanted cells 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 added to 50 ml of growth medium.
When the cells are suspended in a roux flask and 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, and then add 0.2% trypsin solution [4.7 g of Eagle MEM medium (manufactured by Nissui Pharmaceutical Co., Ltd.), 0.6 g of baking soda]
Dissolve g and 1 g of trypsin in 500 ml of distilled water,
Add 10 ml of Millipore filtered solution and treat with trypsin for 2 to 3 minutes at room temperature, 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 a Roux flask. (3) Cell culture and administration of test compound Dispense 1.8 ml of the above cell suspension into a disposable cellar (diameter 35 mm) and place it in a carbon dioxide gas incubator (5% CO ₂ , 95% air) at 37°C for 24 hours. Incubate for 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 with a Millipore filter. (4) How to count the number of cells After administering the test compound, decant the 48-hour shell, discard the supernatant (culture medium), and treat the cells attached to the bottom of the shell with 10 ml of the above 0.2% trypsin solution, which will result in single cells. . Decant this to remove the trypsin solution, make a cell suspension with physiological saline containing 10 mmol phosphate buffer (PH7.0),
Place one or two drops of the sample on a hemocytometer, cover with a cover glass, and count the number of cells under a microscope. The inhibition rate of test cell proliferation was determined by the following formula. Suppression rate (%) = (Number of cells in a test compound-free test sample) - (Test compound-administered cell number) / (Test compound-unadministered cell number) × 100 Next, the active ingredient of the present invention Regarding the toxicity of these compounds, all of them have low molecular structures and are quickly excreted outside the body, so they do not cause any side effects.
In addition, subcutaneous injection and oral administration in mice
All of the LD ₅₀ values are low toxicity compared to other anticancer substances. For example, the LD ₅₀ (mg/Kg) of the above compounds 1, 5, 6, and 7 when administered orally to mice are 500, 500, and 500, respectively. 500, 8000 and above 8000. The present invention will be specifically explained below using formulation examples and test examples. Formulation Example 1 (Injection, Capsule) 30 mg of the above compound 1 was dissolved in 1 g of purified sesame oil and 100 mg of aluminum stearate gel, sealed, sealed with an inert gas such as nitrogen or helium, stored in a cool, dark place, and administered subcutaneously. It is made into an injectable preparation. 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 2 (enteric-coated tablet) β-cyclodextrin (manufactured by Nihon Shokuhin Kako Co., Ltd.)
When 15 g of Compound 7 is mixed in 3000 ml of a saturated aqueous solution of Compound 7 and stirred for 5 hours, the clathrate is precipitated. When this precipitate is dried under reduced pressure, 100 g of the clathrate of Compound 7 is obtained. 1000 enteric-coated tablets each for adults (a) and children (b) were manufactured with the following ingredient composition.

[A]

Main ingredient (clathrate compound of Compound 2 above) 100 (g) Lactose 737 Hydroxypropyl cellulose 3 [B] Cellulose acetate phthalate 80 (g) Hydroxypropyl methyl cellulose phthalate 80 After taking each component of [A] and mixing well. The product was molded into granules according to a conventional method, 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 4 (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 3, 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 The above compounds 1, 2, 3, 4, 5, 6 and 7 were used as test granules of aromatic aldehyde, and the test method was used to cause cancer by the SV ₄₀ oncogenic virus.
Growth inhibition rate (%) of cancer cells W2K・11 in C3H mice
was calculated, and the growth inhibition rate relative activity value of the above test compound was calculated when the growth inhibition rate (%) of benzaldehyde (control) was set as 100. The results shown in Table 3 were obtained.

[Consideration]

As is clear from the results of Test Example 1, all of the above test compounds exhibit activity values equal to or higher than benzaldehyde, when the growth inhibition rate of benzaldehyde, which has excellent anticancer activity, is set as 100. It has been proven that it has excellent anticancer activity. Many of the conventional cancer chemotherapeutic agents have low activity against cancerous cells caused by the SV ₄₀ oncogenic virus compared to transplanted animal cancers, but it is extremely unique that the above compounds act on this and show a suppressive effect. It is noteworthy that In other words, although many of the conventional cytotoxic anticancer drugs show remarkable activity against experimental tumors in animals, that is, transplanted cancers, they still have many problems clinically, and there are very few effective examples. 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 compounds having activity against SV ₄₀ virus-induced cancer are recognized to have very characteristic anticancer activity.

Claims (1)

[Claims] 1. At least one compound selected from the group consisting of O-fluorobenzaldehyde, m-fluorobenzaldehyde, P-fluorobenzaldehyde, O-phthalaldehydic acid, O-phthalaldehyde, m-phthalaldehyde, and P-phthalaldehyde as an active ingredient. An anticancer drug.