JPH0720860B2 - Anti-cancer drug - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a novel antitumor agent containing a surfactant as an active ingredient.

Conventionally, as cancer chemotherapeutic agents, alkylating agents (nitrodien mustards, ethyleneimines, sulfonates), antimetabolites (folate antagonists, purine antagonists,
Pyrimidine antagonist), phytofission venom (corsemide, vinblastine, etc.), antibiotics (sarcomycin, carcinophylline, mitomycin, etc.), hormones (adrenal steroids, male hormones, female hormones), and porphyrin complex acid (murphyrin, copp) Etc. are used.
However, most of them are cytotoxic substances,
Since it has serious side effects, it is strongly desired to develop an antitumor agent having low toxicity and excellent antitumor activity.

Therefore, in view of the above-mentioned gist, the present inventors have searched for a substance having low toxicity and antitumor activity, and as a result of earnest research, as a result, a substance having a surfactant effect causes a change in the membrane and induces cell differentiation. From the expectation that it might be, the differentiation-inducing activity of various surfactants was investigated. As a result, they newly found that they have differentiation-inducing activity on animal tumor cells, and obtained new knowledge that the substance has extremely low toxicity and excellent antitumor activity, and the present invention was obtained. To complete the anticancer drug. The active ingredient of the anticancer agent of the present invention can be an excellent cancer chemotherapeutic agent for warm-blooded animals such as humans, livestock, dogs and cats.

The present invention is a carcinostatic agent comprising, as an active ingredient, at least one compound selected from the group consisting of higher alkyl trimethyl ammonium salts and N-alkyl-4-higher alkyl pyridinium salts.

Specific examples of such compounds include lauryl or oleyltrimethylammonium chloride (Compound (1): Cationogen L (registered trademark)) and N-methyl or ethyl-4-lauryl or oleylpyridinium.
Examples thereof include chloride (compound (2): cationogen H (registered trademark)).

In the present invention, "higher alkyl" means an alkyl group having 9 or more carbon atoms.

The following can be mentioned as an example of these compounds. All of these are known surfactants.

The carcinostatic agent of the present invention can be used either orally or parenterally, and when orally administered, it is administered as soft / hard capsules or tablets, granules, fine granules, powders, and parenterally. Can be administered in subcutaneous or intravenous injections such as aqueous suspensions, oily preparations, infusions and solid or suspension viscous liquids in the form of suppositories so that sustained mucosal absorption can be maintained. .

Formulation of the active ingredient of the present invention, excipients, lubricants, adjuvants,
Further, it can be appropriately carried out by using a pharmaceutically acceptable film-forming substance, a coating aid and the like so as to prepare an enteric-coated preparation if necessary, and specific examples thereof are as follows. In addition, since the active ingredient of the present invention itself exhibits surface activity, it can favorably disintegrate and elute the composition of the present invention.

As an excipient, for example, sucrose, lactose, starch, crystalline cellulose, mannitol, light anhydrous silicic acid, magnesium aluminate, magnesium aluminometasilicate, synthetic aluminum silicate, calcium carbonate, sodium hydrogencarbonate, calcium hydrogenphosphate, carboxymethylcellulose calcium Etc. can be added alone or in combination of two or more.

As the lubricant, for example, magnesium stearate, talc, hardened oil and the like can be added alone or in combination of two or more, and as a flavoring agent and a flavoring agent, salt, satukaline,
Sweeteners such as sugar, mannitol, orange oil licorice extract, citric acid, glucose, menthol, eucalyptus oil, malic acid, etc., flavors, colorants, preservatives and the like may be contained.

Examples of adjuvants such as suspending agents and wetting agents include coconut oil, olive oil, sesame oil, peanut oil, calcium lactate,
Safflower oil, soybean phospholipid, etc. can be contained.

As film-forming substances, cellulose acetate phthalate (CAP) is used as a carbohydrate derivative such as cellulose and saccharides, and methyl acrylate / methacrylic acid copolymer is used as a polyvinyl derivative such as acrylic acid copolymers and dibasic acid monoesters. Examples thereof include polymers and methyl methacrylate / methacrylic acid copolymers.

In addition, when coating the above film-forming substance, a coating-forming agent which is usually used, for example, a plasticizer and various additives for preventing mutual adhesion of chemicals during the coating operation are added to form a film-forming agent. You can improve the properties of the and make the coating operation easier. The active ingredient may be microencapsulated using a film-forming substance and then mixed with an excipient or the like to give a dosage form.

In particular, the compounding ratio in a typical formulation is as follows.

Particularly preferred range Active ingredient 0.1 to 90% by weight 0.3 to 15% by weight Excipient 10 to 99.8% by weight 85 to 99.4% by weight Lubricant 0 to 50% by weight 0 to 20% by weight Film forming substance 0.1 to 50% by weight 0.3 ~ 20 wt% Particularly preferred excipients are lactose, crystalline cellulose, carboxymethylcellulose calcium.

In addition, the dose is an amount sufficient to effectively treat the target tumor and depends on the tumor symptom, the administration route, the dosage form, etc. About 0.01-100 mg / kg body weight (0.01-60 mg / kg for dwarfs)
Body weight), the upper limit is preferably about 50 mg / kg body weight, more preferably about 10 mg / kg body weight, and in the case of parenteral administration, the upper limit is about 10 mg / kg body weight, preferably 5 mg. / kg body weight, more preferably 2 mg / kg body weight is suitable.

Next, a carcinostatic test confirming the carcinostatic activity of the compound of the present invention will be described.

[1] Friend leukemia cell (mouse erythroid leukemia
cell, B8 cells) GIBCO HAM F-12 medium, 15% fetal bovine serum and 60 mg
B8 cells are inoculated at 25 × 10 ⁴ cells / ml into the one to which / of kanamycin is added, and a predetermined amount of the test compound is added thereto (final volume 5 ml).

After culturing in 7.5% CO ₂ at 37 ℃ for 7 days,
The number of cells stained by the benzidine staining method of n), that is, the number of cells adapted to produce hemoglobin due to differentiation into erythrocytes is measured, and the differentiation induction rate is obtained.

[2] Mouse myeloid leukem
IA cell, M1) test GIBCO Eagle MEM medium with 10% horse serum and 60mg /
Mana cells are inoculated to the cells to which kanamycin is added at 5.0 × 10 ⁴ cells / ml, and a predetermined amount of the test compound is added thereto (final volume 5 ml).

After culturing in 7.5% CO ₂ at 37 ° C. for 7 days, lysozyme activity induced by differentiation into phagocytes or granulocytes is examined. The unit of lysozyme activity is Micrococcus lysozyme.
The amount of lysozyme that decreases the absorbance at a wavelength of 450 mμ by 0.001 per minute when measured at pH 6.24 and a temperature of 25 ° C. by using lysozyme as a substrate and a suspension of microbial cells as a substrate.

[3] mouse teratocarcinoma cell
Test for s): One month after the transplantation of teratoma cells from the abdominal cavity of mice to the abdominal cavity was used. Teratoma cells are embryoid bodies (embroid bo
dy) existed as a cell mass and were used without treatment with trypsin. The embryoid bodies obtained by spontaneously submerging in the collected ascites were washed 3 times with a modified Dalbeco's medium or Hanks' solution, then inoculated into a medium containing 10% fetal bovine serum, and a predetermined amount of the test compound was added. CO ₂ 7.5 at ℃
Saturate water vapor containing 8% and incubate in air for 1 week. 0.86 for embryoid bodies obtained by centrifugation (10 minutes at 2000 rpm)
% Nacl solution, naphthol AS-MX phosphate and diazo reagent (Fast Violet B Salt) are added and left at room temperature for 1 hour. This is centrifuged (2000 rpm, 10 minutes) to separate the embryoid body, ethanol is added, and the mixture is left at room temperature for 1 hour. (Undifferentiated cells are colored red).

This is measured for absorption at 535 nm to determine the alkaline phosphatase activity (degree of differentiation induction).

When hexamethylene bisacetamide (HMBA) 5 mM was added (no alkaline phosphatase activity is shown).
Is indicated as "++", the case where HMBA is not added (alkaline phosphatase activity is extremely strong) is indicated as "-", and the degree of differentiation induction is shown in the next step.

++: shows no alkaline phosphatase activity at all.

+: Almost no alkaline phosphatase activity is shown.

±: Shows some alkaline phosphatase activity.

-: Strongly shows alkaline phosphatase activity.

-: Extremely strong alkaline phosphatase activity.

In addition, in the test example described later, it has anti-tumor activity by having a differentiation inducing action.

Hereinafter, the present invention will be specifically described with reference to formulation examples and test examples.

Formulation Example 1 (Injection / Drip) Add 5 g of powdered glucose to contain 10 mg of Compound (1), aseptically distribute to a vial, seal, and seal with an inert gas such as nitrogen or helium in a cool dark place. Save to. Dissolve in ethanol before use, 0.85% physiological saline 100 ml
Is added to give an intravenous injection, and 10 to 100 ml is administered daily by intravenous injection or infusion depending on the condition.

Formulation Example 2 (Injection / Drip) Using 2 mg of Compound (1), a mild intravenous injection is prepared in the same manner as in Formulation Example 1, and 10 to 100 ml is intravenously injected or infused daily depending on the medical condition. Administer.

Formulation Example 3 (Enteric Coated Capsules) 5 g of compound (2), 2.46 g of lactose and 0.04 g of hydroxypropyl cellulose were taken and mixed well, and then formed into granules by a conventional method, dried well and sieved. Separately,
Manufacture granules suitable for heat-sealing packaging. Next, 0.5 g of cellulose acetate phthalate and 0.5 g of hydroxypropylmethyl cellulose phthalate are dissolved to form a coated base material, and the granules are allowed to float and flow, and the base material is coated to form an enteric coated granule. This composition is filled into capsules to produce 100 enteric-coated capsule preparations.

Test Example Using the compound of the present invention, according to the above-mentioned test methods [1], [2] and [3], differentiation induction rate of Friend leukemia cells, lysozyme activity by induction of differentiation of mouse myeloid leukemia cells and mouse teratoma cells When the degree of differentiation induction was examined, a remarkable differentiation inducing activity was observed in the concentration range shown below. The scale of BioAssay is as follows.

(1) Friend leukemia cell (ELC) 1.5% DMSO (+) Hemoglobin 45.1Pg / cell (-) Hemoglobin 12.8Pg / cell (2) Mouse myeloid leukemia cell (MLC) 10 ^-5 M dexamethasone (+) lysozyme 18.9μU / cell (−) lysozyme 5.5 μU / cell (3) Mouse teratoma cell (TER) 5 mM hexamethylenebisacetamide (+) alkaline phosphatase 0.208 IU / ml culture solution (−) alkaline phosphatase 0.349 IU / ml culture Liquid The concentrations (μg / ml) for inducing differentiation of Compounds (1) and (2) were as follows.

As is clear from the results of the above test examples, various surfactants, which are the active ingredients of the present invention, show an action of inducing differentiation into normal cells to cancer cells, and thus have excellent anticancer activity with low toxicity. It was proved to show.

LD ₅₀ value (mg / kg) of the compound of the present invention investigated by intraperitoneally and orally administering the compound of the present invention to mouse 129 strain
Was as follows:

This result indicates that the compound of the present invention can be safely used as an anticancer agent.

Claims (3)

[Claims] 1. An anti-cancer agent comprising as an active ingredient at least one compound selected from the group consisting of higher alkyl trimethyl ammonium salts and N-alkyl-4-higher alkyl pyridinium salts. 2. The first aspect of the present invention is based on a parenteral dosage form.
The anticancer drug according to the item. 3. The anticancer agent according to claim 1, which is in the form of an oral dosage form.