JPH0158161B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to novel anticancer agents, including cinnamaldehyde, α-methyl-cinnamaldehyde, β-furfuracrolein, 2-methoxy-5-acetyl-benzaldehyde, 4-ethoxyisophthalaldehyde, and 4-chloro-6- The present invention provides an anticancer agent containing as an active ingredient at least one compound selected from methyl isophthalaldehyde. Conventionally, cancer chemotherapy agents include alkylating agents (nitrozene mustards, ethyleneimines, sulfonic acid esters), antimetabolites (folate antagonists, purine antagonists, pyrimidine antagonists), and plant fission toxins (colceimide). , vinblastine, etc.),
Antibiotics (sarcomycin, carcinophilin,
mitomycin, etc.), hormones (adrenal steroids, male hormones, female hormones), and porphyrin complex salts (marphyrin, COPP), etc., but in general, the nucleic acid inhibiting effect of anticancer substances affects not only cancer cells but also normal cells. Because of their strong toxicity and serious side effects, it is currently difficult to achieve sufficient effects by using large amounts of drugs such as chemotherapeutic agents for infectious diseases. The present inventor previously developed a new and useful anticancer agent containing benzaldehyde as an active ingredient (Japanese Patent Publication No. 54
(Refer to Publication No. 962), 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 each of the aromatic aldehydes mentioned above has anticancer activity, and these substances may be used for cancer treatment. We have now completed the anticancer agent of the present invention by obtaining new knowledge that it can exert a remarkable effect on cancer. 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 is effective against humans, livestock,
It can be an excellent cancer chemotherapy agent for warm-blooded animals such as dogs and cats. Table 1 shows the physical properties and references of the aromatic aldehyde having anticancer activity used in the present invention.

[Table] (Production Example 1) After refluxing the tetramethylamine salt of 2,4-di-ω-chloromethylethoxybenzene in 60% ethyl alcohol for 5 hours, it was added to ice, and the precipitated crystals were collected by filtration. Recrystallized from ethyl alcohol, melting point 112
Obtain colorless needle-shaped crystals at ~113°C. [nmr( _CDCl3 ), δ1.53(t,3):- _CH3 ,
δ4.28 (q, 2): - CH ₂ -, δ9.86 (s, 1): -
CH0, δ-0.24(s,1):-CH0] 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, in formulating the active ingredient of the present invention, the aromatic aldehyde described above can be made into a water-soluble suspension, oily preparation, etc., into a subcutaneous or intravenous injection preparation according to a conventional method. It can be formulated into dosage forms such as capsules, tablets, and fine granules for oral use. However, among the above compounds, unstable stimulants should be formulated to prevent deterioration and stimulant properties. Removal of is required. That is, the compound must both prevent its autoxidation and improve its stimulant 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. In general, the active ingredient of the present invention can be formulated using surfactants, excipients, lubricants, adjuvants, pharmaceutically acceptable film-forming substances, etc. Specific examples thereof are listed below. For example: 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, 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, 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 substances, in addition to commonly used coating aids such as plasticizers, various additives can be added to prevent the chemicals from adhering to each other during coating operations. properties and make coating operations easier. A typical dosage form of the anticancer agent of the present invention is a dosage form for oral administration, particularly an enteric-coated dosage form. Dosage forms in which the active ingredient is coated with a film-forming substance are particularly preferred because of the relative instability of the active ingredient and because it is enteric-coated. When the active ingredient is in liquid form, it is preferable to form it into a clathrate compound and then mix it with an excipient or the like, or to impregnate it in a porous excipient and then coat it with a film-forming substance. When the active ingredient is in solid form, it is preferable to thoroughly mix it with excipients and the like before coating it with a film-forming substance. Note that a dosage form may be prepared 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. Particularly preferred range Active ingredients 0.1-90% by weight 0.3-15% by weight Excipients 10-99.9 〃 85-99.7 〃 Lubricants 0-50 〃 0-20 〃 Surfactants 0-50 〃 0-20 〃 Film forming Substances 0.1-50 〃 0.3-20 〃 Particularly preferred excipients are lactose, crystalline cellulose, calcium carboxymethyl cellulose. 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 0.5 to 5000 mg/Kg body weight (0.5 to 3000 mg/Kg body weight for dwarfs), and the upper limit is preferably about 200 mg/Kg body weight, more preferably about 50 mg/Kg body weight, and in the case of injections. , its upper limit is about 100 mg/Kg body weight, preferably 50 mg/Kg body weight, more preferably 20 mg/Kg
Appropriate weight. As a result of various tests, it is understood that the anticancer agent of the present invention is effective against human cancer. 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 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~
Corrected 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 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 the mixture becomes cloudy, it is transferred to a Lou 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, then add 10 ml of 0.2% trypsin solution [4.7 g of Eagle MEM medium (manufactured by Nissui Pharmaceutical Co., Ltd.), 0.6 g of baking soda, and 1 g of trypsin dissolved in 500 ml of distilled water and filtered with a Millipore filter]. After additional trypsinization for 2-3 minutes at room temperature, the trypsin solution is decanted. Additionally, 50ml of fresh growth medium
Add to the mixture and wash off the 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 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 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 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. Inhibition rate (%) = 1 - (Number of cells in a shear plate administered with the test compound) / (Number of cells in a shear plate not administered with the test compound) x 100 Next, regarding the toxicity of the active ingredient compound of the present invention, which compound Because of its low molecular structure, it is quickly excreted outside the body and does not cause any side effects.
The LD ₅₀ values of these drugs are also lower than that of other anticancer substances. The present invention will be specifically explained below using formulation examples and test examples. Formulation Example 1 (Injection/Drop) 5 g of powdered glucose was added to contain 500 mg of the above compound 4, and aseptically dispensed 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 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 4 (enteric-coated tablet) β-cyclodextrin (manufactured by Nihon Shokuhin Kako Co., Ltd.)
When 15 g of Compound 1 is mixed in 3000 ml of a saturated aqueous solution of Compound 1 and stirred for 5 hours, the clathrate precipitates. When this precipitate is dried under reduced pressure, 100 g of the clathrate of Compound 1 is obtained. Enteric-coated tablets (1) for adults and 1000 for children (2) were manufactured with the following ingredient composition.

[Table] Loin phthalate Take each component of [A], mix well, pressurize the mixture directly, or knead thoroughly.
The mixture was formed into granules through the screen of an extrusion type granulator, dried sufficiently, and then pressed to produce tablets. Next, it was well dissolved into the molded tablet [B]
The base material is coated to make enteric-coated tablets. About this tablet
That's what it means. ) Disintegration test method, artificial gastric fluid (PH1.2) test of enteric-coated preparations showed that it did not disintegrate even after shaking for 1 hour and shaking it for 1 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] Loose phthalate Enteric-coated granules suitable for capsules were manufactured 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 compounds 1 to 6 as aromatic aldehyde test compounds, cancer cells of C3H mice that were made cancerous by the SV ₄₀ oncogenic virus according to the above test method.
When the inhibition rate (%) of W2K-11 proliferation was calculated, the results shown in Table 2 were obtained.

[Consideration]

As is clear from the results of the above test examples, it was demonstrated that the above test compound had excellent anticancer activity. In addition, the above test compound has an extremely low activity against cancerous cells caused by the SV ₄₀ oncogenic virus, as compared to most conventional cancer chemotherapeutic agents, which have a suppressive effect on 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 has activity against SV ₄₀ virus-induced cancer and has low toxicity, is recognized to have very specific anticancer activity.

Claims (1)

[Claims] 1. Cinnamaldehyde, α-methyl-cinnamaldehyde, β-furfuracrolein, 2-methoxy-5-acetyl-benzaldehyde, 4-ethoxyisophthalaldehyde and 4-chloro-6
-An anticancer drug containing as an active ingredient at least one compound selected from methyl isophthalaldehyde. 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.