JPS6323173B2 - - Google Patents

Description

[Detailed description of the invention]

[Industrial Field of Application] The present invention relates to an anticancer agent. [Background of the Invention] Conventionally, cancer chemotherapy agents include alkylating agents (nitrozene mustards, ethyleneimines, sulfonate esters, nitrosoureas), antimetabolites (methotrexate, ftorafur,
cytosine arabinoside, cyclocytidine, etc.),
Botanical anticancer agents (colcemid, vinblastine, podophyrin, etc.), antibiotics (bleomycin, adriamycin, mitomycin, etc.), hormones (adrenal steroids, male hormones, female hormones), immunostimulants (krestin, picibanil, etc.), and porphyrin complex salts ( Mahfilin,
Anticancer substances (COPP), etc. are used, but in general, anticancer substances act not only on cancer cells but also on normal cells, so they are highly toxic and have serious side effects. At present, it is difficult to achieve sufficient effects through the use of drugs. [Object of the Invention] Therefore, an object of the present invention is to provide a novel anticancer agent that does not have such side effects. [Structure of the Invention] The present invention is an anticancer agent containing a compound represented by the following general formula () or a pharmacologically acceptable acid salt thereof as an active ingredient. R'-φ-CH=N-R () In the formula, φ represents a benzene nucleus or a naphthalene nucleus, and R is a lower alkyl group, a hydroxy lower alkyl group, a lower alkylamino lower alkyl group, a pyridyl group, a piperidylmethyl group, a hydroxy It represents a phenyl group or a benzylideneamino lower alkyl group, and R' represents hydrogen, a lower alkylimino lower alkyl group, a lower alkylamino lower alkylimino lower alkyl group or a hydroxy lower alkylimino lower alkyl group. However, when φ represents a naphthalene nucleus, R represents a lower alkyl group or a hydroxy lower alkyl group, and R' represents hydrogen. Pharmaceutically acceptable acid addition salts of the compound of the above formula () are base salts, sulfates, picrates, citrates, tartrates, fumarates, phthalates, succinates, and adipates. , oxalate, malonate, maleate, malate, etc., and these salts can be easily obtained from the compound of formula () by conventional methods. The known compounds contained in formula () are also not yet known to have anticancer properties.
Any compound of formula () can be considered as a Schiff base derivative of benzaldehyde or terephthalaldehyde. 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. Furthermore, as a result of intensive research into the search for anticancer active substances, it was discovered that the aromatic aldehyde derivatives of formula () each have remarkable anticancer activity. The anticancer agent of the present invention has now been completed based on new findings that the substance 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. Examples of the aromatic aldehyde derivatives of formula () having anticancer activity used in the present invention are shown in Table 1.

【table】

[Table] References and physical property values for the above compounds (1) to (18) (hereinafter, the above compounds are indicated by compound numbers) are shown in Table 2.

【table】

【table】

[Table] Production Example 1 (Production method of compound (1)) Tertiary butylamine in an equimolar amount as benzaldehyde was refluxed for 3 hours while removing azeotropic water in anhydrous benzene, and then the benzene was distilled off under reduced pressure. , Compound (1) is obtained by distillation. Production Example 2 (Production method for compound (2)) Piperidylmethylamine in an equimolar amount as benzaldehyde is refluxed for 3 hours while removing azeotropic water in anhydrous benzene, and then the benzene is distilled off under reduced pressure. Distillation section of 153℃/2mmHg for compound (2)
get as. Production Example 3 (Production method of compound (6)) Compound (6) is obtained by reacting terephthalaldehyde and a large excess of 70% ethylamine aqueous solution at room temperature for 3 days, followed by recrystallization by conventional treatment. Production Example 4 (Production method for compound (7)) After reacting terephthalaldehyde and a large excess of 70% tertiary butylamine aqueous solution at room temperature for 3 days, unreacted substances were distilled off under reduced pressure to obtain compound (7). . Production Example 5 (Production of Compound (8)) Terephthalaldehyde and 2 times the mole of 3-dimethylamino-propylamine were refluxed for 3 hours while removing azeotropic water in anhydrous benzene, and then the benzene and Unreacted substances are distilled off to obtain compound (8). Production Example 6 (Production of Compounds (9) and (10)) Terephthalaldehyde and 2 times the mole of isopropanolamine were refluxed for 3 hours while removing azeotropic water in anhydrous benzene, and then treated in a conventional manner to produce isopropanolamine. Recrystallize from a 1:1 mixed solvent of ether and ethanol to obtain compound (9). The above procedure is repeated using 2-amino-dimethylethanol to obtain compound (10). Production Example 7 (Production method for compounds (11) and (12)) Terephthalaldehyde and 2 times the mole of 2-amino-1-butanol were refluxed in anhydrous benzene for 3 hours while removing azeotropic water, and then refluxed for 3 hours. legal processing,
Recrystallization from ethanol yields compound (11). The above procedure is repeated using ethanolamine to obtain compound (12). Production Example 8 (Production of Compound (11)) Terephthalaldehyde and 2 times the mole of 2-amino-1-propanol were refluxed for 3 hours while removing azeotropic water in anhydrous benzene, and then treated in a conventional manner. Recrystallization from ethanol yields compound (11). 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 orally or by mucosal absorption, and when administered parenterally, When the value is approximately 0.01~
10% by weight is suitable. Furthermore, when formulating the active ingredient of the present invention, the aromatic aldehyde derivative of the above formula () can be made into a water-soluble suspension, oil-based preparation, etc., into a subcutaneous or intravenous injection preparation. In addition to being able to
It can be formulated into dosage forms such as capsules, tablets, and fine granules for oral use. 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, corona 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, methyl acrylate and methacrylate as polyvinyl derivatives such as acrylic acid copolymers and dibasic acid monoesters. Examples include copolymers, methyl methacrylate/methacrylic acid copolymers, and the like. 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, for example, it is appropriate to use a pharmacologically acceptable clathrate compound that utilizes the clathration ability of choleic acid or cyclodextrin.
Formulation using microcapsules is also useful. In addition, in order to give the above-mentioned clathrate compound stability and acid resistance that can withstand long-term retention and to fully maintain its medicinal efficacy, it is possible to formulate a formulation with a pharmaceutically acceptable coating. A stable anticancer drug composition can be obtained. 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 0.5-3000mg. Next, the anticancer activity test 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 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 3-5 ml of sodium hydrogen carbonate solution to correct the pH to 7.1-7.2. Immediately before using the medium, add 10 ml of L-glutamine (2.92 g/100 ml) solution filtered through a Millipore filter. In addition, to preserve the test cells, the final concentration is 10.
% dimethyl sulfoxide. (2) Preparation of transplanted cells Test cells stored in a Jeep freezer (-80℃) were lysed at room temperature, centrifuged at 670× for 5 minutes, the supernatant was discarded, and the precipitated cells were added to the growth medium.
After suspending in 50 ml, transfer to a roux flask and
When cultured at .degree. 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 add 0.2% trypsin solution [Eagle MEM medium (Nippon Pharmaceutical Co., Ltd.)
Dissolve 4.7 g of the product, 0.6 g of sodium bicarbonate, and 1 g of trypsin in 500 ml of distilled water and filter it with 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 with a Millipore filter. (4) How to count the number of cells After administering the test compound, decant the shell for 48 hours, 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. Become. 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 (%) = (Number of cells in a shear plate without administration of the test compound) - (Number of cells in a shear plate administered with a test compound) / (Number of cells in a shear plate without administration of a test compound) x 100 Next, the active ingredient of the present invention Concerning the toxicity of these compounds, all of them are rapidly excreted outside the body due to their low-molecular structure and do not cause any side effects.
Both LD ₅₀ values are low toxicity compared to other anticancer substances, and the above compounds are less toxic when administered orally to mice.
LD ₅₀ (mg/Kg) is shown in Table 3. 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 (12), dispense aseptically into vials, seal them, and fill with inert gas such as nitrogen or helium. Store in a cool, dark place. Before use, add 500 ml of 0.85% physiological saline to make an intravenous injection, and administer for one day.
Administer 10 to 500 ml 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 (11) was used, and administered for one day.
Administer 10 to 500 ml by intravenous injection or drip depending on symptoms. Formulation Example 3 (Injection, Capsule) 30 mg of the above compound (6) 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, and stored in a cool, dark place. 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 Nissui Foods Kako Co., Ltd.)
When 15 g of the above compound (1) is added to 3000 ml of a saturated aqueous solution and mixed and stirred for 5 hours, the clathrate precipitates. When this precipitate is dried under reduced pressure, 100 g of the clathrate compound of the above compound (1) is obtained. It will be done. 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 methylcellulose phthalate 80 Take each component of [A] and mix well. After that, it was formed 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 6 (Enteric-coated capsules) 1000 enteric-coated capsules were manufactured using the following ingredients.

[Table] Cellulose phthalate Enteric-coated granules suitable for capsules were produced 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 Using the above-mentioned compounds (1) to (14), the growth inhibition rate (%) of cancer cells W2K-11 of C3H mice that had become cancerous due to the SV ₄₀ oncogenic virus was calculated according to the test method as shown in Table 3. The results shown are obtained.

〔Effect of the invention〕

As is clear from the results of the test examples, it was demonstrated that the above test compound has excellent anticancer activity. Among the above test compounds, the activity value of Compound (1) is lower than that of the other compounds, but this is because most of the conventional cancer chemotherapy agents are used to treat cancers caused by the SV ₄₀ oncogenic virus, compared to transplanted cancers in animals. It should be noted that the above-mentioned compounds act on these cells and exhibit an inhibitory effect, whereas the activity against these cells is extremely low, and all of the compounds are extremely characteristic in that they have extremely low toxicity. Furthermore, under the above test conditions, compound (7) showed the highest solubility at 10 μg/ml, but it is naturally expected that the above growth inhibition rate would improve if the concentration was further increased. 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 compound, which is active against SV ₄₀ virus-induced cancer and has low toxicity, is recognized to have very specific anticancer activity.

Claims (1)

[Scope of Claims] An anticancer agent containing a compound represented by the following formula () or a pharmacologically acceptable acid salt thereof as an active ingredient. R'-φ-CH=N-R () In the formula, φ represents a benzene nucleus or a naphthalene nucleus, and R is a lower alkyl group, a hydroxy lower alkyl group, a lower alkylamino lower alkyl group, a pyridyl group, a piperidylmethyl group, a hydroxy It represents a phenyl group or a benzylideneamino lower alkyl group, and R' represents hydrogen, a lower alkylimino lower alkyl group, a lower alkylamino lower alkylimino lower alkyl group or a hydroxy lower alkylimino lower alkyl group. However, when φ represents a naphthalene nucleus, R
represents a lower alkyl group or a hydroxy lower alkyl group, and R' represents hydrogen. 2. The anticancer agent according to claim 1, which uses the compound of formula () as a pharmacologically acceptable clathrate compound. 3. The anticancer agent according to claim 1, which is an orally administered agent. 4. The anticancer agent according to claim 3, which contains the active ingredient in an amount of 0.3 to 15.0% by weight. 5. The anticancer agent according to claim 1, which is a parenterally administered agent. 6. The anticancer agent according to claim 5, which contains the active ingredient in an amount of 0.01 to 10% by weight. 7. Claim 1, which takes an enteric-coated dosage form,
The anticancer agent according to any one of Items 3 and 5.