WO2002067940A1

WO2002067940A1 - Application of phenazinone derivatives in preparing antitumor and antimetastases drugs

Info

Publication number: WO2002067940A1
Application number: PCT/CN2002/000121
Authority: WO
Inventors: Qiwang Xu; Junkang Liu; Zetao Yuan
Original assignee: Third Military Medical University, Chinese People's Liberation Army, P.R. Of China
Priority date: 2001-02-28
Filing date: 2002-02-28
Publication date: 2002-09-06
Also published as: CN1372927A; CN1162155C

Abstract

The invention relates to the application of phenazinone derivatives of formula (I) in preparing antitumor and antimetastases drugs. The compounds of formula (I) inhibit the growth of tumor cells by regulating the metabolism of these cells, and block the growth of metastasized cells, and so effect a role of antimetatases. These compounds have significant effects which have been confirmed by the studies in laboratory, and the tests on animals and human beings.

Description

Application of phenazinone derivative in preparing anti-tumor and anti-tumor metastasis medicine Technical field

The present invention relates to the use of phenazinone derivatives in the preparation of anti-tumor and anti-tumor metastatic drugs.

Background technique

Tumors are a common and frequently-occurring disease. Among them, malignant tumors are the most serious type of diseases that currently endanger human health. According to statistics, about 5 million people die from cancer each year, and about 900,000 people die from cancer every year in China. Tumors are new organisms formed by the abnormal proliferation of cells in local tissues under the action of various carcinogenic factors in the body, which often appear as local masses. Tumor cells are transformed from normal cells, but when they become tumor cells, they have abnormal morphology, metabolism, and function.

At present, there are various methods for treating tumors such as surgery, radiation, anticancer drugs, traditional Chinese medicine, and immunization. Among them, surgical treatment is still the most effective treatment method. Chemotherapy has developed rapidly, and currently many types of anticancer drugs such as cytotoxic drugs, antimetabolites, antibiotics, alkaloids, and hormones have appeared. But at present, there are two problems that cannot be solved by anticancer drugs. The first is efficacy. Chemotherapy drugs can only kill a certain percentage of tumor cells. For example, advanced leukemia has 10 ¹² tumor cells, even if a certain drug can kill tumor cells. 99. 99%, there are still 10 ⁸ tumor cells, and clinical recurrence can still occur. The second is that anticancer drugs also have a certain effect on normal cells, especially normal cells that grow and proliferate. Therefore, after administration of drugs, various adverse reactions may occur. Common are ① leukocytes and thrombocytopenia; ② gastrointestinal reactions are nausea, vomiting, diarrhea, oral ulcers, etc .; ③ hair loss; ④ hematuria; ⑤ decreased immune ability; prone to bacterial or fungal infection.

Therefore, there is a need for new antitumor and / or tumor metastatic drugs that are effective in killing tumor cells and have low drug side effects.

Confirm this The present inventors have now surprisingly discovered that some phenazinone derivatives can effectively inhibit tumors without significant toxic and side effects on normal tissues. Therefore, it would be very advantageous to use them to treat tumors and / or tumor metastases. Summary of the invention

Therefore, the present invention relates to the use of a phenazinone derivative of the following formula (I) and a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating tumors and / or tumor metastases

among them, ! ? ^-^ Alkyl or -Cs haloalkyl. In another aspect, the present invention relates to a method of treating tumors and / or tumor metastases, comprising administering to a patient in need thereof a therapeutically effective amount of a halide derivative of formula (I) as defined above, or a pharmaceutically acceptable salt thereof. Pyrosin (R-methyl phenazinone of formula (I)) is a metabolite of P. aeruginosa. Numerous clinical case investigations have shown that P. aeruginosa is rarely found in infectious pathogens of tumor tissues. There were cases of long-term Pseudomonas aeruginosa infection in the lungs, and no cases of metastatic tumors were found. This suggests that certain metabolites of Pseudomonas aeruginosa may have antitumor effects. Pseudomonin was isolated and purified from the metabolites of Pseudomonas aeruginosa, and conventional in vitro tests showed that it had no significant tumoricidal effect. However, fluctuating administration under conditions that regulate metabolism, that is, administration at the beginning of passaging or when tumor cell growth is initiated by regulation of foreign factors, can effectively inhibit tumor cell growth. Surprisingly, the antitumor effect of pyocyanin has been demonstrated by the applicant in animal tests. Tumor cell line Derived from epithelial tissue, connective tissue, and lymphocytes of the blood system. The human trial results show that long-term oral administration of this substance can make the cancer disappear and be used locally, and the cancer tissue will dry up and fall off without affecting the normal tissue.

Therefore, the present invention relates to the application of a phenazinone derivative of the formula (I) and a pharmaceutically acceptable salt thereof to treat various tumors, which can be used as a main active ingredient to prepare an antitumor drug. Because it has been proven that phenazinone derivatives of formula (I) have low toxicity, humans and other mammals can take high doses without significant side effects, and at the same time can effectively inhibit the occurrence and metastasis of tumors and dissipate tumor tissues. The compounds appear to be promising antitumor drugs.

A preferred compound of formula (I) is 1? = Methyl compound of formula (I), ie pyocyanin. Some of the compounds of formula (I) as defined above are commercially available, others can be prepared by conventional methods, for example, by alkylation of compounds of formula (I) where R = H. Pyocyanin can be fermented from P. aeruginosa or other similar strains according to known methods and isolated and purified from the fermented species.

Among the pharmaceutically acceptable salts of phenazinone derivatives of formula (I), mention may be made of those formed with pharmaceutically acceptable acids, such as those formed with inorganic acids, such as hydrochloride, hydrobromide, boron Acid salts, phosphates, sulfates, hydrogen sulfates, and hydrogen phosphates, as well as those formed with organic acids such as citrate, stearic acid, ascorbate, methyl sulfate, naphthalene-2-sulfonate , Picrate, fumarate, maleate, malonate, oxalate, succinate, acetate, tartrate, mesylate, methanylsulfonate, isethionate Salt, _α -ketoglutarate, α-glyceryl phosphate, and glucose-1-phosphate. The compounds of the invention may generally be administered orally, parenterally, sublingually or transdermally, preferably parenterally or orally. In treating tumors and / or tumor metastases according to the method of the present invention, the amount of active ingredient administered depends on the nature and severity of the disease being treated and the weight of the patient. For adults, for systemic preparations, the dosage is 100 mg-3000 mg of a oxazinone derivative of the formula (I) per day. For a topical preparation, the dosage is 500 mg-5000 mg of a hydrazinone derivative of the formula (I) per day.

In the pharmaceutical composition of the present invention for oral, sublingual, subcutaneous, intramuscular, intravenous, transdermal or rectal treatment of the above indications, the active ingredient may be administered to animals and humans in the form of an administration unit, such as It is lyophilized or mixed with conventional pharmaceutical carriers. Suitable dosage unit forms include oral forms, such as oral dispersible tablets, capsules, powders, granules and solutions or suspensions, sublingual and buccal forms, subcutaneous, intramuscular or intravenous Administration forms, topical administration forms and rectal administration forms. Formulations for parenteral and oral administration are preferred, especially injectable solutions (for intravenous, intramuscular, subcutaneous injection), tablets and capsules.

The solid composition in the form of a tablet is prepared by mixing the main active ingredient with a pharmaceutical excipient, such as gelatin, starch, lactose, magnesium stearate, talc, gum arabic, and the like. The tablets may be coated with sucrose or other suitable substances, or they may be treated to have a sustained and delayed effect and to continuously release a predetermined amount of the active ingredient.

Formulations in the form of capsules are obtained by mixing the active ingredient with a diluent and filling the resulting mixture into soft or hard capsules.

Formulations in the form of syrups or pools may contain the active ingredient and preferably a calorie-free sweetener, methyl paraben and propyl paraben as preservatives, and flavoring agents and suitable colorants.

Water-dispersible powders and granules may contain the active ingredient and may be mixed with a dispersing or wetting agent, or a suspending agent such as polyvinylpyrrolidone, and a sweetener or taste modifier.

Rectal administration is performed using suppositories, where the suppositories are made with an adhesive that melts at the temperature of the rectum, such as cocoa butter or polyethylene glycol.

Parenteral administration can be carried out using injectable sterile aqueous, saline, alcoholic solutions or homogeneous suspensions of the compounds of the present invention. Best Mode of the Invention The following test examples use pyocyanin as an example to demonstrate the antitumor and antitumor metastatic activity of the compound of formula (I) of the present invention.

1. In vitro antitumor test of pyocyanin:

1. Cell line: B16 cells

2- Medium: Complete 1640 Medium

3. Method:

3.1 Subculture the cultured B16 cells by trypsinization into culture flasks, and add different doses of chloropyridin after the cells grow, the concentrations are: 1/1 million, 1 / 700,000, 1Λ0,000,000, 1/20 10,000, 1 / 100,000, 1 / 50,000, 1/20, 1 / 10,000, control culture, set daily doses, change the culture medium and drugs for 3 consecutive days. The results are shown in Table (1), indicating green pus Bacteriocin has no killing effect on tumor cells between 1 / 50,000 and 1 / 100,000, but has killing effect only at concentrations above 1/2 000.

3.2 After the cultured B16 cells were digested with trypsin, they were administered at the beginning of a passage, and the concentration was: 1/1 million, 1 / 700,000, 1 / 400,000, 1/20, 1 / The results are shown in Table (2). One is to reduce the concentration of the growing tumor cells and leave them at room temperature. After the shock occurs, the cells are revived and re-administered. The dose setting is the same as above. The results are shown in Table (3). Table 1.B16 cell growth results after continuous administration of different doses of pyocyanin

It can be known from Table 1 that there is no killing effect on tumor cells from 50,000 to V1 million. Growth results of B16 cells given different doses of pyocyanin when activated growth characteristics appear

It can be known from Table 2 that when tumor cells start to grow, psoralen is given again, and it is sensitive, and 1/100 million treatment can play a significant inhibitory effect, and concentrations above 1/2 million show a killing effect. Table 3 B16 cell growth results after administering different doses of pyocyanin when initiating growth characteristics appear

It can be known from Table 3 that when tumor cells start to grow after being regulated by external factors, they are given sensitivity to pyocyanin, and 1/100 million treatment can have a significant inhibitory effect. Extinguishing effect.

Animal tests

Materials and methods

1.1 Animals: Kunming mice, Scad mice 1.2 Tumor cell lines: B16 cells, PG cells

1.3 Methods: B16 cells were injected into the feet of Kunming mice, and they were administered after growing, and were treated with local injection, intravenous injection, oral gavage, and intramuscular injection. The results are shown in the table below.

2. Results and evaluation

As can be seen from the table, local injection of different concentrations of chloropyridin aqueous solution for local treatment of tumors has obvious effects. There are significant differences between the three groups and the control group, and the group I and group II have shown a regression effect after discontinuation. Effect of Oral Administration on B16 Kunming Mouse Transplantation Tumor

Effect of intravenous injection on B16 Kunming mouse transplanted tumors Days of animals Number of tumors (cm ² ) cx¾D Initial size of the control group of the experimental group 15 0.61¾). 38 0.64 37 Treatment 3 days 15 0.68¾). 41 0.71 ¾.36 6 days of treatment 15 0.70 ¾). 29 0.75¾). 32 9 days of treatment 15 0.68¾). 31 0.82 22 12 days of treatment 15 0.60¾). 26 0.90 24 15 days of treatment 15 0.55 19 1.07¾) .21

Observation on the effect of oral administration of pyocyanin on animals

It can be seen from the table that oral administration of pyocyanin has a good inhibitory effect on animal tumors. 3. Pseudomonas toxicity test

Completed by the Third Military Medical University Environmental Hygiene Department.

The experiments include:

1 acute toxicity test

2 Ames experiment

3 Micronucleus test of mouse bone marrow cells

4 mouse sperm deformity experiment

5 mouse testis chromosome aberration experiment

6 dominant lethal experiment

7 Subchronic toxicity (90-day feeding) experiment

8 Traditional Teratogenic Experiments

Experimental conclusion: Pyocyanin is a non-toxic substance 4. Typical cases

1Mr. Chen, 72 years old, went to our center for treatment due to diarrhea, and was found by colonoscopy and pathological biopsy. He was a patient with rectal cancer. After taking Seliqi oral solution (the main functional ingredient is pyocyanin), which was marketed as a health product at that time, the patient reported that the symptoms of diarrhea and bowel sound were controlled after 5 consecutive boxes of use, and the symptoms of tumors were reduced. The tumor did not expand.

2 Guo, 46 years old, rectal cancer was admitted to the Southwest Hospital General Surgery. After rectal cancer resection, the stump continued to develop. We learned that after conducting an antitumor experiment with pyocyanin, we volunteered for a trial. 5ml。 Local injection method, 0.3% concentration injection 0.5ml. Three times in a row, three days after stopping the drug, it was found that the tumor shrank and dried, and continued to be used 5 times, and the local tumor atrophied completely. No tumor metastasis was seen during the administration.

Claims

Rights request

1. Formula (I) phenazinone derivatives and pharmaceutically acceptable salts thereof in the preparation of antitumor and

Application in anti-tumor metastatic medicine

among them, ! ? It is ^-^ alkyl or d-haloalkyl.

2. The use according to claim 1, wherein the derivative is a phenone of the formula (I) wherein R is methyl.

3. The use according to claim 1 or 2, wherein the medicament is an injection preparation suitable for intravenous injection, intramuscular injection, local injection or an external preparation.

4. The use according to claim 3, wherein for systemic preparations, the dosage is 100 mg-3000 mg of phenazinone derivative of formula (I) per day, and for topical preparations, the dosage is 500 mg-5000 mg of formula (I) ) A harazinone derivative.