KR19990036137A - Use of griseofulvin to inhibit cancer growth - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for treating cancer or tumors of a mammal containing griseofulvin, wherein a synergistic chemotherapeutic agent may be used with the griseofulvin, and the griseofulvin alone or other It can be used in combination with other viral agents or synergists to treat viral infections.

Description

Use of griseofulvin to inhibit cancer growth

Cancer, including leukemia, is the leading cause of death in animals and humans. The exact cause of leukemia is unknown, but the link between specific actions, such as smoking or exposure to carcinogens, and the incidence of certain types of leukemia and tumors has been confirmed by many researchers.

Many types of chemotherapeutic agents are known to be effective against cancer, tumors and leukemias, but not all types of cancer and tumor cells respond to the chemotherapeutic agents. Unfortunately, many of these chemotherapy agents also destroy normal cells. The exact mechanism of action of these chemotherapy agents is not always known.

Despite advances in the field of cancer and leukemia treatment, the main treatment methods are still radiation and chemotherapy and bone marrow transplantation. Chemotherapy is particularly attacking active cancer. Such cell breakers or cell proliferation inhibitors work best for cancers with huge growth factors, ie cancers in which cells divide rapidly. To date, hormones, in particular estrogens, progesterone and testosterone, and some antibiotics, alkylating agents, and metabolic antagonists made by various microorganisms form most of the treatments available for oncology. Ideally, cytotoxic agents that have specificity for leukemia, cancer and tumor cells but do not affect normal cells are highly desirable. Unfortunately, no alternative reagents have been found, particularly targeting cells that divide rapidly (hospital cells and normal cells).

Clearly, due to some unique specificities for cancer or leukemia cells, the development of materials targeting these cells has made great progress. Alternatively, a substance that is cytotoxic to leukemia or cancer cells and that exhibits a less irritating effect on normal cells would be preferred. Therefore, it is an object of the present invention to provide a pharmaceutical composition which is effective in treating leukemia and which has little or no effect on normal blood cells.

More specifically, it is an object of the present invention to provide a composition comprising a pharmaceutical carrier and griseofulvin as defined herein, and methods of treating cancer, leukemia and tumors.

The use of the griseofulvin in combination with other chemotherapy agents effective for tumor destruction is a novel method of treatment. The griseofulvin can also be used to treat viral infections in the presence of synergistic factors.

Summary of the Invention

The present invention provides a pharmaceutical composition for the treatment of mammals with leukemia, and particularly warm-blooded animals and humans, comprising a pharmaceutical carrier and an effective amount of griseofulvin. The griseofulvin has the formula (I):

The composition can be used to inhibit the growth of leukemia, tumors and cancer cells in humans or animals by oral, rectal, topical or parenteral, or intravenous administration thereof. The composition does not have a great effect on healthy cells.

In addition, synergistic factors that are chemotherapeutic agents can be used in combination with griseofulvin.

The present invention relates in particular to pharmaceutical compositions useful for the treatment of cancer and tumors in humans and warm-blooded animals.

A. Definition:

As used herein, "comprising" means a number of components that can be used together in a pharmaceutical composition of the present invention. Thus, "consisting essentially of" and "consisting of" are included in "comprising of" above.

As used herein, "pharmaceutically acceptable" component means a component suitable for use in humans and / or animals, without excessive adverse side effects (eg, toxicity, irritation and allergic reactions) corresponding to a moderate profitability / risk ratio.

As used herein, "stable amount and effective amount", when used in the methods of the present invention, provides a desired therapeutic response without excessive adverse side effects (e.g. toxic, irritant and allergic reactions) corresponding to a moderate profitability / risk ratio. It means a sufficient amount of ingredients. The particular “stable amount and effective amount” clearly refers to the particular condition to be treated, the physical condition of the patient, the type of mammal to be treated, the duration of treatment, the nature of the treatment involved (if any), and the particular agent used and It will vary depending on the structure of the compound or derivative thereof.

As used herein, "pharmaceutical addition salt" is an organic or inorganic acid salt of an anti-leukemia compound. These preferred acid addition salts are chloride, bromide, sulfate, nitrate, phosphate, sulfonate, formate, tartarate, maleate, malate, citrate, benzoate, salicylate, ascorbate and the like.

As used herein, “pharmaceutical carrier” is a pharmaceutically acceptable solvent, suspending agent or excipient that delivers the anti-leukemic agent to an animal or human. The carrier may be liquid or solid and is selected according to the intended mode of administration.

As used herein, "cancer" or "leukemia" refers to any type of cancer or tumor or malignant disease that attacks the bone marrow or normal healthy blood cells that produce blood cells found in a mammal.

As used herein, “virus” includes viruses that cause diseases of warm-blooded animals, including HIV, influenza, rhinoviruses, herpes, and the like.

As used herein, “griseofulvin” is 7-chloro-2 ′, 4,6-trimethoxy-6-methylspiro [benzofuran-2- (3H), 1 ′-[2] cyclohexene] -3 Means 4'-dion. It is an antibiotic produced by penicillium griseopulin.

As used herein, “synergism factor” is a substance such as triprolidine and its cis-isomers used in combination with griseofulvin. The synergistic factor may suppress the immune system or improve the efficacy of the drug.

As used herein, "chemotherapeutic agent" includes DNA-interacting agents, metabolic antagonists, tubulin-interacting agents, hormones and other asparaginases or hydroxyureas.

B. Griseofulvin

The griseofulvin has the formula (I):

[Formula I]

It is prepared according to the methods disclosed in US Pat. Nos. 3,069,328 (Hockenhull) (1962) and 3,069,328 (Dorey et al.) (1962).

C. Chemotherapy

Chemotherapy agents are generally classified as DNA-interacting agents, metabolic antagonists, tubulin-interacting agents, hormonal agents and other asparaginases or hydroxyureas. Each group of chemotherapeutic agents may be further classified by activity or type of compound. Chemotherapeutic agents used in combination with griseofulvin include all members of the group. A detailed discussion of chemotherapeutic agents and methods of their administration can be found in Dorr et al., Cancer Chemotherapy Handbook, 2nd ed., P. 15-34, Appleton & Lange (Connecticut, 1994).

Examples of DNA-interacting agents include alkylating agents such as cisplatin, cyclophosphamide, and altamine; DNA chain-breakers such as bleomycin; Intercalated topoisomerase II inhibitors such as dactinomycin and doxorubicin; Non-inserted topoisomerase II inhibitors such as etoposide and teniposide; And the minority DNA binding agent flkamidine.

Alkylating agents form covalent chemical adducts with cellular DNA, RNA and protein molecules and with small amino acids, glutathione and similar chemicals. Generally, such alkylating agents react with nucleophilic atoms in cell constructs such as amino, carboxyl, phosphate, sulfhydryl groups in nucleic acids, proteins, amino acids or glutathione. The mechanism and role of such alkylating agents in the treatment of cancer is not sufficiently known. Typical alkylating agents include nitrogen mustards such as chlorambucil, cyclophosphamide, isopamide, mechloretamine, melphalan, uracil mustard; Aziridine such as thiotepa; Methanesulfonate esters such as busulfan; Nitroso ureas such as carmustine, lomustine, streptozosin; Platinum complexes such as cisplatin and carboplatin; Mitomycin and bioreductive alkylating agents such as procarbazine, dacarbazine and altretamine. DNA chain breakers include bleomycin. DNA topoisomerase II inhibitors include inserts such as amsacrine, dactinomycin, daunorubicin, doxorubicin, idarubicin and mitoxantrone; Non-insert agents such as etoposide and teniposide.

DNA minority binding agent is flcamidine.

Metabolic antagonists interfere with the production of nucleic acids by one or the other of two major mechanisms. Some of these drugs inhibit DNA replication by inhibiting the production of deoxyribonucleoside triphosphate, an intermediate precursor of DNA synthesis. Some of these compounds are sufficiently similar to purine or pyrimidine, which may be their substituents in the assimilation nucleotide pathway. The analog may then be substituted with DNA and RNA instead of its conventional counterpart. Metabolic antagonists useful herein include folic acid antagonists such as methotrexate and trimetrexate; Pyrimidine antagonists such as fluorouracil, fluorodeoxyuridine, CB3717, azacytidine, cytarabine and phloxuridine; Purine antagonists including mercaptopurine, 6-thioguanine, fludarabine, pentostatin; Sugar modified analogues including cycltrabin, fludarabine; There is a ribonucleotide reduction inhibitor comprising hydroxyurea.

Tubulin interactions act by binding proteins that polymerize to form cellular microtubules to specific sites on the tubulin. Microtubules are important cellular structural units. When the interacting agent binds onto a protein, the cells cannot form microtubules. Such tubulin interactions include vincristine and vinblastine, alkaloids and paclitaxel.

Hormones are also useful for the treatment of cancer and tumors. They are hormonally sensitive to tumors and are generally derived from natural sources. These include estrogens, conjugated estrogens and ethynyl estradiols and diethylstilbestol, chlortriacene and idenestrol; Progestins such as hydroxyprogesterone caproate, hydroxyprogesterone and megestrol; Androgens such as testosterone, testosterone propionate, fluoxymesterone, methyltestosterone.

Adrenal corticosteroids are derived from natural adrenal cortisol or hydrocortisone. They are used because of their anti-inflammatory benefits, as well as some ability to inhibit mitosis and stop DNA synthesis. Such compounds include prednisone, dexamethasone, methylprednisolone and prednisolone.

Routine hormones that release hormonal agents, or gonadotropin-releasing hormone antagonists, are mainly used for the treatment of prostate cancer. These include leuprolide acetate and goserelin acetate. They prevent the biosynthesis of steroids in the test.

Antihormonal antigens include antiestrogens such as tamoshifen, antiandrogens such as flutamide, and antiadrenal agents such as mitotaines and aminoglutetimides.

Hydroxyurea is thought to act mainly in the process of inhibition of the enzyme ribonucleotide reductase.

Asparaginase is an enzyme that converts asparagine into nonfunctional aspartic acid and thus blocks intratumoral protein synthesis.

D. Rising Factor

"Elevation factor" may be any substance that improves or increases the efficacy of a pharmaceutical composition, or acts as an immunosuppressant. One such synergistic factor is triprolidine and its cis-isomers used in combination with chemotherapeutic agents and griseofulvin. Triprolidine is disclosed in US Pat. No. 5,114,951 (1992). Other synergistic factors include procodazole, 1H-benzimidazole-2-propanoic acid; [β- (2-benzimidazole) propionic acid; 2- (2-carboxyethyl) benzimidazole; Propazole]. Procodazole is a nonspecific active immunoprotectant against viral and bacterial infections and can be used with the compositions claimed herein. The compound is effective for treating cancer, tumors, leukemias and viral infections with griseopulin alone or in combination with chemotherapy.

Propionic acid and salts and esters thereof may also be used in combination with the pharmaceutical compositions claimed herein.

Antioxidant vitamins such as vitamins A, C and E, and beta-carotene can be added to these compositions.

E. Dosage

Suitable dosages are described in the methods of the invention. The type and dosage of the compound and the carrier vary widely depending on the type of warm-blooded animal or human, body weight, and type of cancer or tumor or viral infection to be treated. Generally, in the case of griseofulvin or chemotherapy, the dosage is suitably about 1 to about 8,000 mg per kg of body weight. Preferably, 15 to about 5,000 mg per kg body weight is used. In general, human dosages are less than small warm blooded mammals such as mice. The dosage unit may contain a single compound or a mixture of this and other compounds or other cancer inhibiting compounds. Dosage units may also contain diluents, extenders, carriers, liposomes, and the like. The unit may be in solid or gel form, such as pills, tablets, capsules or the like, or liquid form suitable for oral, rectal, topical, intravenous or parenteral administration or injection into or around the bone marrow. The range and ratio between griseofulvin and chemotherapy depends on the type of cancer or tumor to be treated and the type of particular chemotherapy.

F. Dosage Form

Chemotherapy agents, griseofulvin, and optionally synergistic factors are usually mixed with pharmaceutically acceptable carriers. The carrier may be solid or liquid, the type of which is generally selected according to the dosage form used. The active agent can be administered as a tablet or capsule, such as agglomerated powder, in liposome form, or in liquid form. Examples of suitable solid carriers include lactose, sucrose, gelatin and agar. Capsules or tablets can be easily formulated and made easy to swallow or chew. Other solid forms include granules and bulk powders. Tablets may contain suitable binders, lubricants, diluents, disintegrants, colorants, flavors, flow-inducing agents, and melting agents. Examples of suitable liquid dosage forms include solutions or suspensions from water, pharmaceutically acceptable fats and oils, alcohols or other organic solvents (including esters), emulsions, syrups or elixirs, suspensions, solutions and / or suspensions reconstituted from non-bubble granules, And foam preparations reconstituted from foam granules. Such liquid dosage forms may contain, for example, suitable solvents, preservatives, emulsifiers, suspending agents, diluents, sweetening agents, thickening agents and melting agents. Oral dosage forms optionally contain flavoring and coloring agents. Parenteral and intravenous dosage forms will also contain minerals and other substances that make them suitable for the type of injection or delivery system selected.

Specific examples of pharmaceutically acceptable carriers and excipients that can be used to prepare oral dosage forms of the invention are disclosed in US Pat. No. 3,903,297 to Robert (September 2, 1975). Techniques and compositions for preparing dosage forms useful in the present invention are disclosed in the following references: 7 Modern Pharmaceutics, Chapters 9 and 10 (Banker & Rhodes, Editor, 1979); Pharmaceutical Dosage Forms: Tablets (1981) (Lieberman et al.); And Introduction to Pharmaceutical Dosage Forms, 2nd Edition (1976) (Ansel).

G. Treatment Methods

The treatment method can be any method effective for the treatment of the specific type of cancer or tumor to be treated. Treatment can be oral, rectal, topical, parenteral or intravenous administration or injection into a tumor or cancer. The method of administering the effective amount also depends on the leukemia, cancer, tumor or virus to be treated. Parenteral treatment by intravenous, subcutaneous or intramuscular administration of griocevin, formulated with a suitable carrier, an additional cancer inhibiting compound, or a compound or diluent that facilitates administration is a preferred method for administering the compound to warm-blooded animals Can be.

In addition to the use of chemotherapy agents and synergists, griseofulvin can be combined with fungicides, herbicides or other antiviral agents. Preferred preparations and fungicides are carbendazim, fluoroconazole, benoyl, glyphosate and propicodazole.

Example 1

For acute HIV in an in vitro model, griseofulvin inhibited 98% viral replication at 10 μg / ml with a therapeutic index of 5.3. The known HIV drug AZT also inhibited 98% viral replication at 1 μg / ml with a therapeutic index of 12,500. The therapeutic index is the ratio between the toxic drug dose and the effective drug dose.

Example 2

In an in vivo study of mice for leukemia (P388), griseofulvin was 156% at a dose of 4,000 mg / kg, 188% at a dose of 5,000 mg / kg, and 218 at a dose of 6,000 mg / kg. An increase in survival time for% untreated control was shown.

Example 3

In an in vivo study of mice for melanoma (B16), griseofulvin was 165% at a dose of 4,000 mg / kg, 179% at a dose of 5,000 mg / kg, and 6,000 mg / kg. An increase in survival time was shown for the 201% untreated control. Cytosan at 300 mg / kg showed an increased survival of 192%.

Example 4

For in vitro population screening for rhinovirus, type A-1, cell line WI-38, griseofulvin was effective at 100 μg / ml. A positive control was A-36683, (S, S) -1,2-bis (5-methoxy-2-benzimidazolyl) -1,2-ethanediol from Abbott Company. The A-36683 had a therapeutic index of 1,000 to 3,200. Griseofulvin had a therapeutic index of 1-2 (see Applied Microbiology, 23, No. 1, 113-116 (1972)).

Example 5

Solid tumors cut from patients were chopped to a size of 2 to 5 mm and then placed in McCoy's medium 5A + 10% heat-inactivated newborn calf serum + 1% penicillin / streptomycin. Within 4 hours, these solid tumors were mechanically separated with scissors and No. Squeezed through a 100 stainless steel mesh, 25 gauge needles, and then washed with McCoy's medium as described above. By standard techniques, ascites, pleural, pericardial fluid and bone marrow were obtained. The fluid or bone marrow was placed in a sterile container containing 10 units of preservative-free heparin per ml of harmful fluid or bone marrow. After centrifugation at 150 x g for 10 minutes, the cells were harvested and washed with McCoy's medium + 10% heat-inactivated calf serum. The viability of the cell suspension was measured with trypan blue on a hemocytometer.

The cells to be cloned were treated with 15% heat-inactivated horse serum, penicillin (100 units / ml), streptomycin (2 mg / ml), glutamine (2 mM), insulin (3 units / ml), asparagine ( 0.6 mg / ml) and 0.3% agar in abundant CMRL 1066 supplemented with HEPES buffer (2 mM). For the continuous exposure test, each compound was added to the mixture. To prevent the growth of fibroblasts, cells were placed in a 35 mm Petri dish on the top of the agar above the bottom of the agar. Three plates were prepared for each data point. The plates were placed in a 37 ° C. incubator and then recovered on day 14 to count the number of colonies in each plate. By comparing the number of colonies formed in three compound-treated plates (defined as 50 cells) with the number of colonies formed in three control plates, the percentage of colonies surviving at the concentration of the compound can be assessed. Three positive control plates were used to measure survival. 200 μg / ml orthosodium vanadate was used as a positive control. The test was evaluated when the colonies in the positive control were 30% or less compared to the untreated control.

In single dose experiments of griseofulvin, concentrations of 0.5 and 5.0 μg / ml were not effective against tumors in the test (0/1). In a continuous exposure experiment of griseofulvin, a concentration of 50.0 μg / ml was effective for colon, lung, non-small cell and ovarian cancer. 5/6 and above all had a survival rate of 50% or less.

Claims (10)

A pharmaceutical composition for the treatment of cancer and tumors containing 1 to about 8,000 mg of the pharmaceutical carrier per kg of body weight. A pharmaceutical composition according to claim 1 which contains a synergistic factor. The pharmaceutical composition according to claim 1 or 2, comprising a pharmaceutically acceptable carrier, a stable amount and an effective amount of griseofulvin and a chemotherapeutic agent. The method of claim 1, wherein the chemotherapeutic agent is selected from the group consisting of DNA-interacting agents, metabolic antagonists, tubulin-interacting agents, hormones, asparaginases or hydroxyureas. Pharmaceutical composition. The method of claim 1, 2 or 4, wherein the chemotherapeutic agent is asparaginase, hydroxyurea, cisplatin, cyclophosphamide, altretamine, bleomycin, dactinomycin, doxorubicin, A pharmaceutical composition selected from the group consisting of etoposide, teniposide and flcamidine. 5. The method of claim 1, wherein the chemotherapeutic agent is methotrexate, fluorouracil, fluorodeoxyuridine, CB3717, azacytidine, cytarabine, phloxuridine, mercaptopurine , 6-thioguanine, fludarabine, pentostatin, cycltrabin and fludarabine. A method of treating cancer or tumors in a warm blooded mammal comprising administering a composition according to any one of claims 1 to 6. A method of treating viral infection in a warm blooded mammal comprising administering a composition according to any one of claims 1 to 6. A unit dosage composition for the treatment of a viral infection in an animal or human containing from 1 to about 8,000 mg of the pharmaceutical carrier per kg of body weight. A unit dosage composition for the treatment of cancer or tumors in an animal or human containing from 1 to about 8,000 mg of the pharmaceutical carrier per kg of body weight.