SK46999A3 - Pharmaceutical composition - Google Patents

Abstract

A method of treating HIV or other viral infections by administering a herbicide or fungicide or derivative thereof to an animal or human. The fungicides or herbicides can be used in conjunction with other treatments, e.g. with AZT or protease inhibitors for the treatment of HIV. For example, thiabendazole and chloropropham have been shown to quickly reduce the level of virus production from cell populations chronically infected with HIV-1 and the antiviral effect is maintained with continued compound exposure. This reduction of virus production occurs at concentrations which are non toxic to the host cell and which have no effect on the syntheses of cellular DNA, RNA and protein. Further, chronically infected cells treated for prolonged periods of time with thiabendazole and chloropropham were not super-infected with HIV. A method for inhibiting the growth of tumors and cancers in mammals comprising administering a herbicidal or fungicidal derivative is also disclosed herein. The fungicides or herbicides can be used in conjunction with other treatments, e.g. taxol for the treatment of breast cancer. Potentiators can also be included in the herbicidal or fungicidal composition. This method is particularly effective when the cancer or virus is an animal cell genetically modified by plant or fungus genetic material. A chemotherapeutic agent can also be administered first to significantly reduce the size of the cancer and then the treatment with the herbicide or fungicide is used. These methods are particularly effective when the cancer or virus is a mutated cell comprising plant or fungal genetic material.

Description

The invention relates to a method of inhibiting the growth of viruses, in particular HIV, cancer and tumors in humans and warm-blooded animals, and comprises administering herbicidal and fungicidal agents. Tumor size decreases, malignant tumor growth slows, and viral replication is inhibited. This treatment is particularly effective when the virus or cancer is the result of a mutated cell, i.

An animal cell that is mutated by the incorporation of genetic material from plants, fungi and fungi.

BACKGROUND OF THE INVENTION

HIV and other viral infections are among the leading causes of death. HIV is a disease in which a replicating virus attacks the body's immune system. HIV virus cannot be easily destroyed even if the mechanism that prevents replication of the virus in host cells works well. Herpes simplex is another viral infection that can be treated very difficult. A method of treating these diseases and other viral infections is highly desirable.

Surprisingly, it has been found that fungicides, herbicides,

fungal inhibitors and derivatives thereof can inhibit viral replication. The fungicides and herbicides may be used in conjunction with other treatments, e.g. AZT, 3TC or protease inhibitors for the treatment of HIV. This HIV treatment is significantly effective in the treatment of chronically infected cells and cells that do not appear to develop resistance to treatment. For example, it has been shown that thiabendazole and chlorpropham rapidly reduce the level of virus production in the cell population of chronically infected HIV-1 and the antiviral effect persists after subsequent exposure (up to one year). Further, this significant, substantially complete suppression of viral production persists in the chronically inability to induce the experiment. In contrast, resistant strains a

Malignant tumors of both animals and humans, malignant tumors, the emergence of infected cells up to 80 days after the drug is removed and this vaccine effect is at the same level as if both substances were present. This effect has not been described in the HIV treatment literature to date. Reduction of virus production occurs at concentrations that are nontoxic to host cells and that have no effect on cellular DNA, RNA, and protein synthesis. Furthermore, chronically infected cells treated for a long time with tiabendazole and chlorpropham were not superinfected with HIV. Further, a resistant strain is important for these agents after one-year protease inhibitors of the week induce all RT inhibitors to induce resistant strains for one to two months.

are among the main causes of death The exact cause is unknown but many scientists describe different types of tumors in connection with various activities, such as smoking, carcinogenic exposure. However, it is known that tumor cells are abnormal cells that may sleep for some time and subsequently grow rapidly. Many tumor cells are considered immortal because they do not die as normal animal and human cells, but continue to spontaneously propagate.

It is obvious that the breakthrough will be the development of substances with unique specificity that will be directed against tumor cells. Otherwise, it is desirable that substances having a cytotoxic effect on tumor cells show only a slight effect on normal cells.

It is known that plant and genetically altered by the introduction of a different cell species. These genetically altered cells have DNA or a portion of the genes or genetic material of one cell incorporated into a new cell. These new cells exhibit features of both cells, but are no longer identified as entirely plant or animal cells. Such altered or animal cells may be genetic material from abnormal animal cells may be rapidly growing and do not respond as normal cells by a regulatory mechanism.

The organism has various mechanisms to filter out and eliminate molds, fungi and pollen substances. If ingested,

The digestive system can secrete or detoxify them in reasonable amounts. The nose, lungs and skin contain active mechanisms for filtration of foreign substances. However, there will always be plants, fungi and fungi that penetrate the body's defense mechanisms and reach the bloodstream, lymphatic system, or through the lungs and skin membranes. These substances can penetrate and alter animal cells. The altered cells may be formed by combining plant cells with animal cells formed by combining pollen with an animal cell. Pel can be absorbed by breathing, as the lung tissue and other cells of the nose, mouth and throat are in permanent contact with plant substances. Other cells can be formed by combining fungal and fungal substances with animal cell genetic material in the same way.

Abnormal or altered cells that contain genetic material of plant and animal cells or fungi and animal cells are altered cells in the environment. The organism normally rejects them by generating antibodies, but if the cells exhibit characteristics of both animal and plant cells or fungi, including fungi, the antibodies may not function in the same way. This allows cells to grow due to their abnormality, which is called cancer or tumor growth, and also includes so-called cancer growth. liquid tumors, e.g. leukemia.

Viruses also have the ability to attack cells and multiply in cell mass. These new cells are different from the two original cells. The viruses themselves mutate and change due to their interaction with fungi, molds and plant material, especially pollen. These viruses can also be destroyed or their replication inhibited by treatment with fugicides, mold inhibitors and herbicides. Interestingly, some of these substances also have anthelmintic activity.

Interestingly, 15% of malignant tumors are caused by viruses. Geographically speaking, the presence of fungi, especially those found in grasses, and the use of herbicides and fungicides to control the growth or proliferation of these substances have been found to correlate with the occurrence of malignant tumors at these sites. The more the growth of fungi and plants was inhibited by the use of herbicides and fungicides, the less malignant tumors occurred in the area.

It is an object of the invention to provide a method of treating HIV wherein the production of the virus is suppressed. The method comprises administering a safe and effective amount of a herbicide or fungicide alone or in combination with other HIV drugs, such as e.g. AZT or 3TC.

It is another object of the present invention to provide an antiviral treatment which comprises administering a safe and effective amount of a viral growth inhibitory agent which also has the ability to destroy viruses, particularly altered and altered cells. These substances are herbicides that can destroy plant cells and fungicides that are effective against fungi, including fungi. These substances are also administered in conjunction with a multiplier, such as an anti-inflammatory drug, or vitamins, including antioxidant vitamins.

It is a further object of the invention to provide an anticancer treatment which comprises administering a safe and effective amount of a tumor attenuating agent that has the ability to destroy tumor cells, particularly altered cells. These substances are herbicides that can destroy plant cells and fungicides that are effective against fungi, including fungi. These agents are administered with the chemotherapeutic agent either simultaneously or sequentially and / or in conjunction with a multiplier.

These and other objects will be apparent from the following detailed description of the invention.

SUMMARY OF THE INVENTION

A method of treating HIV and other viral infections and malignant tumors in mammals and especially warm-blooded animals is a claim. The method comprises administering a safe and effective amount of a herbicide or fungicide or derivatives thereof that significantly reduces tumor mass or inhibits replication of viruses and tumor cells. The herbicidal agent and its derivatives are effective in the treatment of malignant tumors or viruses, which are genetically altered animal cells resulting from the association of a plant cell with an animal cell. Fungicides and derivatives thereof are effective when the cancer or virus is an animal cell that contains genetic material derived from fungi or fungi.

These components are used to inhibit the growth of malignant and other tumors in humans or animals. An effective amount is administered either by mouth, rectum, topically or parenterally, intravenously or by injection directly into the tumor. Multipliers are also used in this mixture. The mixture is used in conjunction with other treatments, either concurrently or sequentially.

DETAILED DESCRIPTION OF THE INVENTION

A. Definitions

As used herein, a component is understood to mean various components that can be combined into the pharmaceutical agent of the invention. Accordingly, the term consisting of essentially consisting of and consisting of.

As used herein, the terms mutant cell and environmentally altered cell mean an animal cell that is genetically altered by joining genetic material, i. DNA and RNA segments from plant cells or fungal cells with animal cell genetic material to produce a new, genetically modified cell that is neither plant, fungal nor animal, but which is a viable parasite that persists in the host cell. As such a cell grows and multiplies, it becomes a malignant cell in the host animal.

►

As used herein, the terms "pharmaceutically acceptable substance" means a substance that is suitable for use in human and / or animal without unnecessary side effects (eg toxicity, irritation, allergic response), with an acceptable benefit / risk ratio.

As used herein, the term safe and effective dose refers to an amount of a substance that is suitable for achieving a therapeutic response without undue side effects (e.g., toxicity, irritation, allergic response) with an acceptable benefit / risk ratio when used according to the method of the invention . The term safe and effective dose varies depending on the factors of the particular treatment conditions, the physical condition of the patient, the species of mammal being treated, the time of treatment, the type of concurrent treatment (if any) and the exact structure of the substances and their derivatives.

As used herein, the term pharmaceutical salts means a salt of herbicidal or fungicidal substances and derivatives thereof with an organic or inorganic acid. Popular acid salts include chlorides, bromides, sulfates, nitrates, phosphates, sulfonates, formates, tartrates, maleates, malate, citrates, benzoates, salicylates, ascorbates and the like.

As used herein, the term derivative means a chemically modified derivative of a fungicidal or herbicidal substance that is more soluble or more easily metabolizable, but does not have a significantly altered herbicidal or fungicidal effect. For example, the addition of a hydroxyl group or other hydrophilic group improves solubility and absorption in the body and in most cases does not affect the functionality of the substance. Those skilled in the art will readily determine these substances.

As used herein, the term pharmaceutical carrier is a pharmaceutically acceptable solvent, suspending agent or vehicle, including liposomes, for delivery of the anti-cancer agent to an animal or human. The carrier may be liquid or solid and its choice will depend on the intended route of administration.

As used herein, the term cancer includes k

Also all malignant tumors and neoplasms in mammals, including tumors and leukemia.

As used herein, the term chemotherapeutic agent includes DNA agents, antimetabolites, tubulin agents, hormonal agents and others, such as asparaginase or hydroxyurea.

As used herein, viruses include viruses that cause diseases (viral infections) in humans and other warm-blooded animals, such as HIV virus, herpes, influenza virus and rhinoviruses.

In this document you multiply the expression used! are substances such as triprolidine and its cis-isomer and procodazole which are used in combination with a chemotherapeutic agent and herbicidal or fungicidal agents.

As used herein, the term "significantly decreases" means a significant reduction in tumor mass. Usually it is below 50% of the original mass, but above all the reduction of the mass to undetectable size.

As used herein, fungicides means substances that are effective in inhibiting fungal growth or in controlling fungi. Fungal inhibitors are included in the term fungicides, since fungi are fungi.

As used herein, the term herbicides means substances which are effective in inhibiting the growth of plants or those which destroy plant cells.

B. Fungicides

Any number of fungicides and their derivatives and pharmaceutically acceptable salts are used. The particular fungicide is selected for both safety and efficacy in preventing fungal growth. A wider range of fungicides is preferred. Some effective fungicides are shown below.

1. Benzimidazole compounds

Bezimidazole derivatives are known for their antifungal activity. Surprisingly, it has been found that these agents can cause apoptosis of the cancer cell line. Apoptosis is a specific cell death that differs from necrosis. Most tumor cells can live indefinitely. Tumor cells are referred to as immortalized cell lines. Therefore, the ability to induce apoptosis is very important.

The compounds of formula I have the following structure:

(I) wherein X is hydrogen, halogen, alkyl of less than 7 carbon atoms or alkoxy of less than 7 carbon atoms, n is an integer of less than 4, Y is hydrogen, chlorine, nitro, methyl or ethyl, and R 2 is hydrogen, CONHR 3 and R 3 is alkyl of less than 7 carbon atoms, preferably butyl or isobutyl or an alkyl group of 1 to 8 carbon atoms, and R ₂ is NHCOOR 1 wherein R 1 is an aliphatic hydrocarbon of less than 7 carbon atoms, preferably an alkyl group of less than 7 carbon atoms.

A preferred compound has the general formula II:

wherein R is hydrogen, CONHR 3 and R 3 is alkyl of less than 7 carbon atoms, preferably butyl or isobutyl or alkyl of 1 to 8 carbon atoms or non-toxic, pharmaceutically acceptable salts of both organic and inorganic acids.

Most preferred compounds are methyl (butylcarbamoyl) -2-benzimidazolecarbamate, 2-methoxycarbonylaminobenzimidazole and compounds wherein Y is chlorine and X is hydrogen. These compounds are prepared as described in U.S. Pat. 3,738,995 Adams et al., June 12, 1973.

2. Thiabenzimidazole compounds

Thiabendazole has been found to be effective in the treatment of chronic HIV, especially when suppression of viral production has been demonstrated without developing cell resistance.

Compounds of formula III:

wherein X is hydrogen, halogen, alkyl of less than 7 carbon atoms or alkoxy of less than 7 carbon atoms, n is a positive integer less than 4, Y is hydrogen, chloro, nitro, methyl or ethyl, and R is hydrogen, CONHR ₃ , wherein R 3 is an alkyl group of less than 7 carbon atoms, preferably butyl or isobutyl or an alkyl group of 1 to 8 carbon atoms, and R 2 is thiazolyl. Preferred is a formulation of formula IV:

wherein R is hydrogen and R 2 is 4-thiazolyl or non-toxic pharmaceutically acceptable salts of both organic and inorganic acids. The most preferred compounds are 2- (4-thiazolyl) benzimidazole. Thiabendazole is also an anthelmintic agent.

Thiazolyl derivatives are prepared according to the method of Brown et al., J. Am. Chem. Soc., 83, 1764 (1961) and Grenda et al., J. Org. Chem., 30, 259 (1965).

3. Substituted benzimidazole derivatives

More soluble benzimidazole compounds are useful in the present invention. While these compounds are not known for herbicidal or fungicidal use, they are believed to have effects against HIV, malignant tumors and other viral infections. These derivatives are compounds of formula V:

(V) wherein R 1 is selected from the group consisting of hydrogen, carboxyl (-CO ₂ H), hydroxyl, amino, or esters (-CO ₂ R '), wherein R' is selected from the group consisting of alkoxy, haloalkyl, alkenyl and cycloalkyl, wherein the alkyl groups have 1 to 8 carbon atoms or CH ₃ CH ₂ (OCH ₂ CH ₂ ) _n - or CH ₃ CH ₂ CH ₂ (OCH ₂ CH ₂ CH ₂ ) or (CH ₃ ) ₂ CH-- -a compound of formula VI:

and (OCH _{(CH3) CH2) n} ---, n is 1 to 3. The alkyl groups are preferably straight-chain. The halogen is preferably substituted on the terminal carbon and the halogen is chlorine. Preferred are those cycloalkyl groups having 3 to 6 carbon atoms. Cycloalkyl groups also include those substituted on the alkyl chain, 2-cyclopropylethyl, cyclopropylmethyl, 2-cyclopropyl propyl or 2-cyclopropylpropyl or cyclohexylmethyl. Preferred are compounds of formulas VII to XI:

(IX),

(X),

and pharmaceutically acceptable salts thereof (XI)

4th

1Η-1,2,4-triazole derivatives

1H-1,2,4-triazole derivatives are known for their fungicidal activity. They are systemic substances used to prevent and control fungi. These are compounds of formula XII:

• I

I o o I '/

CH, -C-Ar (XII) wherein Z is an alkylene selected from the group consisting of CH ₂ CH ₂ 'CH ₂ CH ₂ CH ₂ -, -CH (CH ₃₎ CH (CH ₃₎ - and -CH ₂ -CH (alkyl) wherein said alkyl contains 1 to 10 carbon atoms, Ar is a member selected from the group consisting of phenyl, substituted phenyl, thienyl, halothienyl, naphthyl, and fluorenyl, wherein substituted phenyl means that on the phenyl radical are 1 to 3 substituents selected independently from the group consisting of halogen, lower alkyl, lower polyalkoxy, cyano and nitro. The therapeutically active acid salts of the aforementioned compound (I) are also included within the scope of this invention.

As used in the above definition of Z, the term alkyl includes straight and branched chain hydrocarbon radicals containing from 1 to 10 carbon atoms, such as e.g. methyl, ethyl, 1-methylethyl, propyl, 1,1-dimethylethyl, butyl, pentyl, hexyl, heptyl, octyl, decyl and the like. As used herein, the term lower alkyl means straight and branched chain saturated hydrocarbons having 1 to 6 carbon atoms, such as e.g. methyl, ethyl, propyl, 1-methylethyl, butyl, 1,1-dimethylethyl, pentyl, hexyl and im-like alkyls. The term halo generally refers to halogen atoms having an atomic mass of less than 127, i. fluorine, chlorine, bromine and iodine.

Also used herein are their pharmaceutically acceptable salts of both organic and inorganic acids.

Preferred derivatives include:

1- [2- (2,4-Dichloro-phenyl) -1,3-dioxolan-2-ylmethyl] -1H-1,2,4-triazole;

1- [2- (2,4-Dichloro-phenyl) -4-methyl-1,3-dioxolan-2-ylmethyl] -1H-1,2,4-triazole;

I

1- [2- (2,4-Dichloro-phenyl) -4-ethyl-1,3-dioxolan-2-ylmethyl] -1H-1,2,4-triazole;

1- [2- (2,4-Dichlorophenyl) -4-propyl-1,3-dioxolan-2-ylmethyl] -1H-1,2,4-triazole;

1- [2- (2,4-Dichlorophenyl) -4-pentyl-1,3-dioxolan-2-ylmethyl] -1H1,2,4-triazole and therapeutically active salts of their acids.

These compounds are prepared as described in U.S. Pat. No. 4,079,062 Van Reet et al., Mar. 14, 1999.

5. 1,3-Bis-triazolyl-2-propanol derivatives 1,3-bis-triazolyl-2-propanol derivatives are known for their fungicidal activity. They are systemic substances used to prevent and control fungi. These are compounds of formula XIII:

OH

C-CH, -C-CH, R

(XIII) wherein R 1 is optionally substituted alkyl, cycloalkyl (e.g. cyclopentyl or cyclohexyl), aryl or haloaryl (e.g. phenyl or 2,4-dichlorophenyl) or aralkyl (e.g. benzyl); and their salts and metal complexes and ethers or esters, or non-toxic, pharmaceutically acceptable acid salts, both organic and inorganic. In particular, bis-triazole derivatives such as 2- (2,4-dichlorophenyl) -1,3-bis (1H-1,2,4-triazol-1-yl) propan-2-ol and the corresponding 2- and 2- 4-chlorophenyl analogs and 2,4-dichlorophenyl analogs. Preferred is the composition of 2- (2,4-dichlorophenyl) -1,3-bis (1H-1,2,4-triazol-114-yl) propan-2-ol and their pharmaceutically acceptable acid salts, both organic and inorganic.

The compounds are prepared as described in U.S. Pat. No. 4,404,216, Richardson, Sep. 13, 1983; No. 2,078,719A, issued Jan. 27, 1982 (both to Imperial Chemical Industries, Ltd.).

6. Griseofulvin

Griseofulvin is a compound of formula XIV:

It is prepared as described in U.S. Pat. 30169,328, Hockenhull (1962) and U.S. Pat. No. 3,069,328, Dorey et al. (1962).

C. Herbicides

Any number of herbicides and their derivatives and pharmaceutically acceptable salts are used. Preferred are the herbicides described below:

N-chlorophenylcarbamates and N-chlorophenylthiocarbamates

N-chlorophenylcarbamates and N-chlorophenylthiocarbamates are known for their herbicidal activity. They are systemic substances used to prevent and control certain plants and weeds. Systemic herbicides differ from other herbicides in their ability to be absorbed and permeated by the plant. This systemic capability is not an essential requirement for the compounds of this invention.

Chlorpropham, isopropyl N- (3-chlorophenyl) carbamate appears to be particularly effective in the treatment of HIV.

These are compounds of formula XV:

O-R'- "

- H (XV) wherein n is 1 to 3, X is oxygen or sulfur and R is selected from the group consisting of hydrogen, lower alkyl, lower alkenyl, cyclohexyl, C 1 -C 8 phenalkyl, and a pharmaceutically acceptable salt of these compounds .

Preferred are those compounds in which R is C 1 -C 4 alkyl, preferably isopropyl and X is oxygen, n is 1 and the chloro group is in the 3-position of the phenyl group. Ν-3-Chlorophenylcarbamate is preferably used.

These compounds are prepared as described in U.S. Pat. No. 2,695,225 to Witman (1954) and U.S. Pat. No. 2,734,911, Strain (1956).

2. N-phosphonoglycines

N-phosphonoglycine derivatives are known for their herbicidal activity. They are systemic substances used to prevent and control certain plants and weeds.

These are compounds of formula XVI:

ABOUT

H

X-CH2-C

R

I n-ch ₂ o

II

P-OY (XVI), wherein X is selected from the group consisting of hydroxy, thioyl, alkoxy or chloroxy of up to 12 carbon atoms: lower alkenoxy, cyclohexyloxy, morpholino, pyrrolidinyl, piperidino and NHR '; Y and Z are independently selected from hydrogen and lower alkyl, R is selected from the group consisting of hydrogen, formyl, acetyl, benzoyl, nitrobenzoyl and chlorinated benzoyl, R 'is selected from the group consisting of hydrogen, lower alkyl and lower alkenyl, cyclohexyl, phenalkyl of up to 8 carbon atoms, phenyl, chlorinated phenyl and anisyl, and certain salts of these compounds selected from the group consisting of metals of groups I and II having an atomic number of up to 30, hydrochloride, pyridine, ammonia, lower aliphatic amine hydrocarbons, lower aminoalkanols and aniline.

Most preferred are compounds of formula XVII:

O CH) O

II I li

HO-C-CH ₂ -N-CH ₂ -P- (OH) ₂ (XVII),

Preferably, lower alkylamine salts are used, in particular zopropylamine salts.

These compounds are prepared as described in U.S. Pat. No. 3,794,758, Franz, Dec. 10, 1974.

The pharmaceutical composition for the treatment of mammals, especially warm-blooded animals and man, consists of a pharmaceutical carrier and an effective amount of an antitumor agent selected from the group consisting of (1) N-phosphonoglycine derivatives and (2) N-chlorophenylcarbamates or are also useful herein N-chlorophenylthiocarbamates.

D. HIV drugs

HIV is treated with two general classes of drugs, reverse transcriptase inhibitors and protease inhibitors. AZT and 3TC are widely used to treat acute HIV infection. Herbicidal and fungicidal agents and their derivatives are used in combination with AZT or 3TC in the treatment of acute HIV infection. They do not interfere with AZT activity.

Other HIV and antiviral agents in combination are also used in the present invention. These include reverse transcriptase inhibitors and protease inhibitors.

The drugs are used either simultaneously or administered sequentially with fungicidal or herbicidal agents and derivatives thereof.

E. Chemotherapeutic agents * I, ·

Fungicides and herbicides may be administered with chemotherapeutic agents. They are administered either sequentially, the chemotherapeutic agent is used to reduce the tumor, followed by treatment with the herbicide, fungicide or derivatives thereof, or the two are administered simultaneously.

Chemotherapeutic agents generally belong to classes of DNA interacting agents, antimetabolites, tubulin interacting agents, hormonal agents, and others such as asparaginase or hydroxyurea. Each of these groups of chemotherapeutic agents is further subdivided by activity type or compound. Chemotherapeutic agents that are used sequentially in combination with herbicidal or fungicidal agents include primary agents from the group of DNA interacting agents, antimetabolites, tubulin interacting agents, antimetabolites, and tubulin interacting agents. For a detailed description of chemotherapeutic agents and their mode of administration, see Dorr et al., Cancer Chemotherapy Handbook, 2nd Edition, p. 15-34, Appleton & Lange (Connecticut, 1994), incorporated herein by reference.

In order to reduce tumor mass or stop tumor cell growth, the chemotherapeutic agent must prevent cell replication but have the ability to protect the cell itself. Such are substances that are primarily active on DNA, e.g. cisplatin and tubulin agents.

DNA agents include alkylating agents, such as e.g. cisplatin, cyclophosphamide, altretamine, DNA breakers such as e.g. bleomycin, interstitial topoisomerase II inhibitors, such as e.g. etoposide and teniposide, and plicamycin.

Alkylating agents form covalent bonds with cellular DNA, RNA and protein molecules and lower amino acids, glutathione and similar chemical compounds. Generally, these alkylating agents react with a nucleophilic particle in the cell mass, such as amino groups, carboxy groups, phosphate, sulfohydril groups in nucleic acids, proteins, amino acids, or glutathione. The mechanism and role of these alkylating agents in the treatment of malignant diseases has not yet been elucidated. Typical alkylating agents include:

nitrogen mustards such as e.g. chlorambucil, cyclophosphamide, isophosphamide, mechloretamine, melphalan and uracil mustard;

aziridines, e.g. thiotepa;

methanesulfonic acid esters, e.g. busulfan; nitrosoureas such as e.g. carmustine, lomustine, and streptozocin;

platinum complexes, e.g. cisplatin and carboplatin; bioreductive alkylating agents, e.g. mitomycin, procarbazine, dacarbazine and altretamine;

DNA chain damaging agents, e.g. bleomycin; DNA topoisomerase II inhibitors, including the following:

substances with intercalating effect such as e.g. amsacrine, dactinomycin, daunorubicin, doxorubicin, idarubicin, and mitoxantrone;

non-intercalating agents such as e.g. etoposide and teniposide;

DNA binding agents, such as e.g. plicamycin.

Antimetabolites interfere with the production of nucleic acids by one or two major mechanisms. Some of the drugs block the formation of deoxyribonucleoside triphosphates, which are the immediate precursors for DNA synthesis and thus inhibit DNA replication. Some compounds are sufficiently similar to purines and pyrimidines to replace them in nucleotide formation. These similar substances can then replace them in DNA and RNA instead of their normal counterparts. Here, the antimetabolites used include:

folic acid antagonists such as e.g. methotrexate and trimetrexate;

pyrimidine antagonists such as e.g. fluorouracil, fluorodeoxyuridine, CB3717, azacytidine, cytarabine, and floxuridine;

purine antagonists such as e.g. mercaptopurine, 6-thiogvanine, fludarabine, pentostatin;

modified sugar analogs including cyctrabine, fludarabine;

ribonucleotide reductase inhibitors including hydroxyurea.

Tubulin actives bind to specific sites of tubulin, a protein that forms cellular microtubules by polymerization. Microtubules are critical structural cell units. When the agent binds to the protein, the cell cannot form microtubules. Tubulin actives include vincristine, vinblastine, both alkaloids and paclitaxel.

The adrenal corticosteroids are derived from adrenal cortisol or hydrocortisone. They are used for their anti-inflammatory effects, as well as the ability to stop mitotic division and stop DNA synthesis. These compounds include prednisone, dexamethasone, methylprednisolone and prednisolone.

The hydroxyurea primarily acts by inhibiting the enzyme ribonucleotide reductase.

Asparagenase is an enzyme that converts asparagine to a dysfunctional aspartic acid and thereby blocks protein synthesis in the tumor.

Hormonal agents and luteinizing hormones are usually not used to significantly reduce tumor mass. However, they can be used in conjunction with chemotherapeutic agents or herbicidal or fungicidal agents or derivatives thereof.

Hormone blocking agents are also useful in the treatment of cancer. They are used in hormone-alterable tumors and are usually obtained from natural sources.

They include:

estrogens, conjugated estrogens and ethinylestradiol and diethylstilbestrol, chlorotrianizene, idenestrol;

progestins such as e.g. hydroxyprogesterone caproate, medroxyprogesterone and megestrol;

androgens such as e.g. testosterone, testosterone propionate, fluoxymesterone, methyltestosterone.

Luteinizing hormone releasing hormone antagonists or gonadotropins are used primarily for the treatment of prostate cancer. They include leuprolide acetate and goserelin acetate. These substances prevent the synthesis of steroids in the gonads.

Antihormonal agents include: antiestrogenic agents, e.g. tamoxifen, antiandrogenic agents, e.g. flutamide; and antiadrenal agents, e.g. mitotane and aminoglutethimide.

F. Potentiators

Potentiators can be any substances that improve and enhance the efficacy of pharmaceutical agents and / or act on the immune system. One such potentiator is triprolidine and its cis-isomer, which are used in combination with chemotherapeutic agents and fungicides or herbicides. Triprolidine is described in U.S. Pat. 5,114,951 (1992).

Another potentiator is procodazole, 1H-benzimidazole-2-propanoic acid, [beta- (2-benzimidazole) propionic acid,

2- (2-carboxyethyl) benzimidazole; propazol). Procodazole is a non-specific effective immunoprotective agent against viral and bacterial infections and can be used in the compounds claimed herein.

Potentiators can improve the efficacy of the herbicidal and fungicidal compounds and can be used in a safe and effective amount. These combinations may be administered to the patient or animal by mouth, rectum, topically or parenterally.

Antioxidant vitamins such as ascorbic acid, beta-carotene, vitamin A and vitamin E may be administered with the compounds of the invention.

G. Dosage

Any suitable dose may be administered by the method of the invention. The type of substances, carriers and amounts vary depending on the species of warm-blooded animal or human, body weight, and the virus or malignancy being treated. The extent and proportion of the fungicidal or herbicidal agents used and their derivatives and / or chemotherapeutic agents depend on the type of substance and the type of malignancy being treated. In general, a dosage of from 2 milligrams (mg) per kilogram (kg) of body weight to 4000 mg per kg of body weight is suitable for the herbicidal or fungicidal substances and their derivatives. Higher doses up to 6000 mg / kg may also be used. Preferably, fungicidal or herbicidal agents are also used in doses of from 15 mg / kg to 3000 mg / kg of body weight. For chemotherapeutic agents, lower dosages are suitable, i. from 0.01 mg / kg body weight to 400 mg / kg body weight. Generally, doses for humans are lower than for small warm-blooded mammals such as mice. The dosage unit consists of either a single agent or a mixture thereof with other agents that suppress cancers.

Any suitable dose may be administered by the method of the invention in the treatment of HIV infection. The type of substances and carriers and the amounts will vary depending on the species of warm-blooded animal or human and body weight. The range and proportion of fungicidal or herbicidal agents used and their derivatives and agents for the treatment of HIV infection depends on the type of agent. In general, a dosage of from 0.2 mg / kg body weight to 4000 mg / kg body weight is suitable for fungicidal or herbicidal substances and derivatives thereof. Higher doses up to 6000 mg / kg may also be used. Preferably, fungicidal or herbicidal agents are also used in doses of from 20 mg / kg to 3000 mg / kg of body weight. For chemotherapeutic agents, lower dosages are suitable, i. from 0.01 mg / kg body weight to 400 mg / kg body weight. In general, human doses are lower than those for small warm-blooded mammals such as mice. The dosage unit consists of either a single agent or a mixture thereof with other agents for the treatment of HIV infection.

H.

Pharmaceutical forms

The herbicides or fungicides and their derivatives are usually admixed with a pharmaceutically acceptable carrier. The carrier may be solid or liquid or in the form of liposomal particles. The form is generally administered as a single drug, in the form of a tablet or capsule, in liquid form. An example.

selects based on the method

The active agent is administered in a liposome, as a powder or suitable solid carriers are lactose, sucrose, gelatin and agar. Capsules or tablets are easy to manufacture and easy to swallow or chew. Other solid forms include granules or bulk powders. Tablets may contain suitable binders, lubricating oils, diluents, grinding agents, coloring agents, flavors, flowing agents and release agents. Examples of suitable liquid dosage forms are aqueous solutions or suspensions, pharmaceutically acceptable fats and oils, alcohols and other organic solvents, including esters, emulsions, syrups or elixirs, suspensions, solutions and / or suspensions of non-effervescent granules, and effervescent effervescent granules. Liquid dosage forms may contain e.g. suitable solvents, preservatives, emulsifiers, suspending agents, diluents, sweeteners, thickeners and release agents. Oral dosage forms include optionally flavoring and coloring agents. Parenteral and intravenous forms also contain minerals and other agents to be compatible with the selected type of injection or target system.

Specific examples of the pharmaceutically acceptable carriers and ointment bases of the present invention which are used in the manufacture of oral dosage forms are described in U.S. Pat. 3,903,297, Robert, Sep. 2, 1975. The procedures and compounds for making the dosage forms used in this invention are described in the following references: 7 Modern Pharmaceutics, Chapters 9 and 10 Banker & Rhodes, Editors, (1979), Lieberman et al. Pharmaceutical Dosage Forms: Tablets (1981); and Ansel, Introduction to Pharmaceutical Dosage Forms 2 ^nd Edition (197 6).

I. Method of treatment

The method of treatment may be any suitable method that is effective in treating individual viral diseases, malignancies and benign tumors. Treatment may be by oral, rectal, topical, parenteral or intravenous administration and the like. The mode of administration of the effective amount varies and depends on the type of tumor or viral disease being treated. It is believed that parenteral treatment by intravenous, subcutaneous or intramuscular administration of herbicidal or fungicidal agents is accomplished using a suitable carrier. The additional anti-viral agents may be used together with the herbicide or fungicide as well as the additional anti-tumor agents may be combined in the treatment. Solvents are used to facilitate the deposition or administration of substances to warm-blooded animals.

Doses of herbicidal or fungicidal agents that are used to treat viral infections are administered for 7 to 21 days or longer to inhibit growth or kill viruses. In the case of chronic infections, the substances may be administered for a longer period, up to several years.

For the treatment of acute viral infections or HIV infection, a fungicidal agent is administered after AZT treatment or in combination with other HIV treatment. These medicines are administered in a sequential regimen in which the HIV virus is first weakened in the body and the subsequent herbicidal or fungicidal agent prevents it from multiplying. AZT treatment may be continued during treatment with a herbicidal or fungicidal agent. When the disease is at an early stage, the herbicidal or fungicidal agent is administered to prevent the growth or growth of the virus and thereby slow the progression of the disease.

In the treatment of malignant diseases, a herbicidal or fungicidal substance is preferably first administered in order to significantly reduce the tumor mass. This takes 3 to 14 days. Tumor reduction or tumor cell levels reach 50% of the original size. Radiation therapy is used in combination with herbicidal or fungicidal treatments.

Once the tumor is reduced, the herbicidal or fungicidal agent is administered. Being relatively safe, they can be administered for 14 to 365 days to maintain the efficacy of reducing regrowth malignancy.

The following examples are illustrative and are not intended to limit the invention.

Benomyl is methyl 1- (butylcarbamoyl) -2-benzimidazolecarbamate

Carbendazim is methyl 2-benzimidazole carbamate

Thiabendazole is 2- (4-thiazolyl) -1H-benzimidazole

DETAILED DESCRIPTION OF THE INVENTION

Example 1 HIV testing

HIV replication studies

Chlorpropham and thiabendazole have been studied in chronically infected with HIV. These cell populations contain incorporated copies of the HIV genome and thereby produce HIV at relatively high levels (CEM-SK1, U937-SK1 and H9-SK1 from Frederick Research Center, Maryland) or are secretly infected and produce virus only after stimulation with phorbol esters, tumor necrosis factor or IL 6 (U1 and ACH2). Virus production was reduced in all cell lines tested and the substances did not stimulate viral production in covertly infected cells. A decrease in viral production was quantitated by the activity of reverse transcriptase, p24 supernatant, as well as intracellular p24, which determines that the substance inhibits viral production at the replication stage, preferably over intracellular protein production.

Quantification of the infectivity of virions produced by the infected cells demonstrates a reduced number of infectious virions concurrently with a decrease in RT or p24 supernatant. This proves that the substances reduce the amount of virus produced but not the quality of the virions. A decrease in viral production in chronically infected cells was found at concentrations that are non-toxic to the target cells. Thiabendazole inhibits viral production at concentrations greater than 1 to 10 pg / ml, while chlorpropham inhibits viral production at concentrations greater than 0.25 pg / ml.

Toxicity in chronically infected cells was similar to that found in uninfected cells. Evaluation of the action of chlorpropham and thiabendazole on chronically infected cells was performed by evaluating the incorporation of thymidine (DNA), uridine (RNA) and leucine (protein) into cell macromolecules. Inhibition of synthesis of cellular macromolecules concurrently with the toxicity of two substances has not been demonstrated

¹ I expected, at lower nontoxic concentrations found to inhibit virus production from the chronically infected cells.

After 28 days of treatment of chronically infected cells with these agents, the toxicity of the agents to the target cells appears similar to both infected and uninfected cells. Preferably, the substances do not kill cells infected with HIV. The decrease in the level of viral production was stable and was found at concentrations higher than 10 gg / ml for thiabendazole and higher than 1 gg / ml for chlorpropham.

These results suggest that chlorpropham and thiabendazole can rapidly reduce the level of viral production of populations chronically infected with HIV-1 and are maintained with prolonged exposure to cellular antiviral activity.

This reduction in viral production occurs at concentrations that are nontoxic to the host cell and that have no effect on the synthesis of cellular DNA, RNA, and proteins.

Viral resistance studies

Cells chronically infected with HIV were cultured in the presence of 1 pg / ml thiabendazole during the first month, 5 pg / ml during the second month, 10 gg / ml during the 3 month, 20 and 40 gg / ml during the 4 month and 80 gg / ml for 5 to 6 months. Chlorpropham was used at concentrations of 1,2,4,8 and 16 gg / ml for each of the six months. At the end of each month, cells were scored for viral production by comparison with cells chronically infected without treatment with these agents. For each of the six months of experimental treatment, there was no change in the antiviral effect of the substances and their toxicity remained the same. The substances persisted against HIV and did not rapidly develop resistance through the selection of resistant viruses or by adapting the cells to prevent drug-induced toxicity. Viral production remains completely suppressed in cultures treated with 40 and 80 gg / ml thiabendazole and 8 and 16 pg / ml chlorpropham, respectively.

Recurrence of viral production in chronically infected cells previously treated

Chronically infected cells were treated for a prolonged period, then the substance was washed out and the cells were further cultured to determine if and when viral production would occur. Cultures that completely eliminated viral production were used in these assays. These cultures included chronically infected cells, cultured in the presence of 20, 40 and 80 pg / ml thiabendazole and 4, 8 and 16 gg / ml chlorpropham respectively. Within 4 days, virus production was resumed if lower concentrations of both substances (20 gg / ml and 4 µg / ml) were used. Virus production was resumed at 40 µg / ml tiabendazole on day 12 and at 8 µg / ml chlorpropham on day 54. At higher concentrations, virus production was observed at approximately 70 days.

Infectiousness of cells treated with chlorpropham and thiabendazole

Cells were eluted from cells that had been pretreated with chlorpropham and thiabendazole for a longer period of time and were used as target cells. These cells were divided into three populations and designated Group 1, 2 or 3. Group 1 was treated with the compound for 24 hours (at the same concentration as used in the extended treatment phase), then the substance was washed and the cells were cultured in the presence of infectious virus. and fresh substance. Group 2 was pre-treated for 24 hours, then the substance was washed out and the cells were cultured only in the presence of infectious virus. Group 3 was cultured in both the pre-treatment and infectious phases only in fresh medium (virus or substance free). Viral production in cell populations was identical, regardless of culture conditions. These results demonstrate that chronically infected cells treated for prolonged periods were not superinfected with HIV.

Complementary studies of chronic HIV infection

U1 cells chronically infected with HIV-1 were obtained from cells of the promonocytic line U937 acutely infected with HIV-1. ACH-2 cells chronically infected with HIV-1 were obtained from the T-cell line A3.01, acutely infected with HIV-1.

These cells were cultured in medium and phorbol ester, PMA. PMA causes cells (both U1 and ACH-2) to activate but do not divide, but also cause U-1 cells to differentiate. This leads to a decrease in the number of cells in cultures treated with PMA compared to the cultures in the medium or the cultures themselves. The viability of the cells after being treated with the enhancers is measured.

Both cell lines subsequently produce small amounts of HIV-1. The ACH-2 cell lines tend to produce more HIV-1 than U1 cells, as evidenced by the p-24 ELISA. When both cell lines are cultured in the presence of PMA, there is an increase in the amount of HIV-1 production measured by the β-24 antigen by ELISA.

In addition, the number of cells that produce HIV-1 positive mRNA per microscopic field is measured. The comparison is made by adhering to the same number of cells on a glass slide for each drug concentration (10 x 10 ⁶ cells / ml).

These cells were treated with test samples.

'J

f. '

Thiabendazole at 60 µg / ml suppressed replication in HIV monocytes by 74% and HIV replication in T cells increased by 26%. A positive control was AZT, which at a concentration of 1 µg / ml suppressed replication in HIV monocytes by 98% and suppressed HIV replication in T cells by 60%. The therapeutic index (TI), the ratio of the toxic dose of the drug to the effective dose of the drug, is 2.8 for tiabendazole, as opposed to 12,500 for AZT.

Model of acute HIV infection

In an acute in vitro HIV infection model, griseofulvin inhibited viral replication by 98% at a concentration of 10 µg / ml with a therapeutic index of 5.3. AZT, a known HIV drug, also attenuated viral replication by 98% at a concentration of 1 gg / ml with a therapeutic index of 12,500. The therapeutic index is the ratio of the toxic dose of the drug to the effective dose of the drug.

Example 2

Tests of colon, breast and lung tumor cells

The following cell culture assays were performed to verify the toxicity of herbicidal or fungicidal agents on human colon, breast and lung tumor cells. Cell viability was assessed by tumor shrinkage using MTT (3- [4,5-dimethylthiazol-2-yl] -2,5-diphenyltetrazolium bromide). The MTT assay is a known assay for measuring cell viability.

Colon tumor cells (HT29 from the American Type Culture Collection (ATCC) and breast tumor cells (MXI from ATCC cell lines) were cultured in Eagle's minimal baseline medium with 10% fetal bovine serum.

Tumor cells were passaged and seeded into culture flasks at the desired cell densities.

¹ I

The culture medium was decanted and the cell coating was washed twice with phosphate buffer (PBS). Cells were digested with trypsin and split prior to seeding into culture flasks. Unless otherwise specified, cultures were incubated at 37 ± 1 ° C in a humidified atmosphere with 5 ± 1% carbon dioxide. Cultures were allowed to incubate until they coincided with 50-80%.

Cell cultures were cultured incompletely until they completely blended. The medium was aspirated from the flask and the cell coatings were washed twice with PBS. Then trypsin solution was added to each flask to cover the cell coatings. After 30 to 60 seconds, the solution was removed and the flasks were incubated at room temperature for 2 to 6 minutes. As soon as 90% of the cells were released, growth medium was added. Cells were then removed by spreading and transferred to a sterile centrifuge tube. The concentration of cells in the suspension was determined, suitable dilutions were made to achieve a density of 5000 cells / ml. Cells were incomplete cultured into the desired wells of 96-well bioassay plates (200 μΐ cell suspension / well). PBS was added to all remaining wells to ensure moisture. Plates were then allowed to incubate overnight before starting treatment.

Each dose of test substance was tested in four-well culture with 100 μΐ of each dilution. Additional wells were added to the wells designated as solvent controls

100 μΐ of the methanol control, an additional 100 μΐ of treatment medium was added to the negative controls. PBS was added to the remaining wells that were not treated with test substance or medium. The plates were then incubated for 5 days.

At the end of the 5th incubation day, each dose group was examined microscopically to assess toxicity. A 0.5 mg / ml MTT solution was prepared in the treatment medium and the solution was filtered through a 0.45 μιη filter

I, so as to remove undissolved crystals. The medium was poured from the wells of the plates. Immediately thereafter, 2000 μΐ of filtered MTT solution was added to each well, in addition to the two untreated blank wells. 200 μΐ of treatment medium was added to the two blind wells. The plates were then returned to the incubator for 3 hours. After incubation, MTT medium was decanted from the wells containing it. Excess medium was added to each well and the plates were allowed to shake at room temperature for 2 hours.

Absorbance at 550 nm (OD550) was measured with a VMax sensor from Molecular Devices (Menlo Park, CA).

The mean absorbance of the OD550 wells with solvent controls, all wells with test substance solution, and all blank wells and positive controls was calculated. The OD550 absorbance mean of the blank wells was subtracted from the diameter of the wells of the solvent controls and from the wells of the test substance solution to obtain the corresponding OD550 mean.

corrected OD ₅₅₀ of test substance solution% control ----------------------------------------- ------- x 100 corrected mean OD ₅₅₀ solvent control

Dose sensitivity curves were plotted as semi-logarithmic plots with% control on the linear axis and test compound concentration on the logarithmic axis. EC 50 was read from the graph for each test substance.

Separate sensitivities were made for the methanol test data for test substances administered in methanol.

Adriamycin was used as a positive control. In all cases, it was more than one to two times more toxic than any of the test substances. Adriamycin is one of the more potent substances currently used with significant side effects. Peak plasma concentrations of other fully active chemotherapeutic agents are 10 to 50 times higher than that of adiamycin.

EC50 refers to the concentration at which half of the cells are killed.

The following tables show test results for individual fungicides and herbicides. The effect of these substances on normal healthy cells is also determined.

Table no. 1

Test substance The result EC-50 / Ppm / HT29 HT29 MXI MXI A549 A549 adriamycin 0.03 0,006 0.02 0,001 0.03 0,009 benomyl 0,742 0,747 1.42 2.42 0,980 1.02 carbendazim 0,621 0, 662 0,829 0,856 0,856 0,836

The following results were obtained for normal healthy cells:

Table no. 2

Test substance EC -50 cells bronchi cells corneal fibro blasts. benomyl .728 0,682 3.26 2.4 3.24 2.81 carbendazim 0,320 0,506 0,752 0,822 1.52 1.42 adriamycin 0,015 0.0020 0.0035 0.0093 0,065 0.10

Lung tumor cells (A549), breast tumor cells (MCF-7) and intestinal tumor cells (HT-29) were used in the present study. Thiabendazole, a systemic fungicide, effectively killed these cells. Table no. 3 is a summary of the results.

Table no. 3

Concentration (ppm) Optical density A-549 MC F-7 HT-29 0-control '0,600 '0,245' 0,398 173 0,007 0,007 0,005 35 0,411 0,025 0,011 17.3 0, 851 0,258 0,204 3.46 1.12 0,466 0,713 0.87 1.32 0,507 0,852

The results of the chlorpropham tests are shown in Tables 1 and 2. 4a no. 5

Table no. 4

Test substance The result EC-50 / ppm or pg / ml / HT29 HT29 MXI MXI A549 A549 adriamycin 0,003 0, 006 0.02 0,001 0.03 0,009 chlorpropham 13.3 11.4 91.8 108 12.6 92.5

The following results are shown for normal healthy cells.

Table no. 5

Test substance EC -50 bronchial cells corneal cells fibro blasts chlóprofam 0,002 > 15.2 3.9 13.0 > 152 64.2 adriamycin 0,015 0.0020 0.0035 0.0093 0,065 0.10

The results of the tests with glyphosphate are shown in Tables no. 6 and 7

Table no. 6

Test substance The result EC-50 / Ppm / HT29 HT29 MXI MXI A549 A, 54 9 adriamycin 0,003 0,006 0.02 0,001 0, 03 0,009 glyphosate 5.41 3.73 36.5 14.6 25.9 22.3

The following results are shown for normal healthy cells.

Table no. 7

Test substance EC -50 bronchial cells corneal cells fibro blasts glyphosate 1.59 3.54 3.09 3.21 86.1 35.8 adriamycin 0,015 0.0020 0.0035 0.0093 0,065 0.10

A mixture of chlorpropam and glyphosphate was also tested. The results are shown in Tables. 8 and no. 9th

Table no. 8

Test substance The result EC-50 / Ppm / HT29 HT29 MXI MXI A549 A549 adriamycin 0,003 0,006 0.02 0,001 0.03 0.009. chlorpropham 13.3 11.4 91.8 108 12.6 92.5 glyphosate 5.41 3.73 36.5 14.6 25.9 22.3 1: 1 mixture * 1.96 1.61 9.70 8.78 10.8 10.1

a mixture of chlorpropham and glyphosphate®

The following results are valid for normal healthy cells.

Table no. 9

Test substance 1 _f EC -50 bronchial cells corneal cells fibro blasts chlóprofam 0,002 > 15.2 3.9 13,0 ' > 152 64.2 glyphosate 1.59 3.54 3.09 3.21 86.1 35.8 1: 1 mixture * 0,001 0,497 0,242 0,286 129 5.95 adriamycin 0,015 0.0020 0.0035 0.0093 0,065 0.10

* a mixture of chlorpropham and glyphosphate®

In table no. 10 shows the results of the propiconazole test.

Table no. 10

Test substance Result EC-50 (ppm) HT29 MXI A549 adriamycin 0.00639 0.00078 0.00373 propiconazole 0.0331 0.0284 0,113

Leukemia studies

Mice were selected randomly and divided into treatment groups. Five groups were infected with leukemia. These diseased animals were dosed for five days, skipped for the next days, then dosed for an additional five days and skipped for another three days, then dosed for five days and skipped for another two days. This irregular dose and dose missed variation is not an ideal regimen, but the results appear to be positive for carbendazim ⁽ TM ⁾ . One group of mice was treated with cytoxane ^(TM) , 2- [bis (2-chloroethyl) -amino-1-oxo-2aza-5-oxophosphoridine, the control group received canola oil, and the third group was treated with different levels of carbendazim ^(TM) , methyl (butylcarbamoyl) -2-benzimidazole carbamate. A non-treatment control group was also used. Carbendazim ^(TM) was administered in three different doses, 4000 mg / kg, 2500 mg / kg and 1000 mg / kg. Cytoxan ^(TM) was administered at 125 mg / kg. After 8 days, a group of untreated mice lost one mouse, after 10 days 8 mice died, and on day 11 all 10 mice died. Mice from the cytoxane ( ^{TM) group} survived more than 21 days. In the higher dose group of carbendazim ^(TM> one mouse died on day 14, two died on day 15, 16, and 17, and one on day 20, 21, and 22. The average number of days for this group was 17, 3. In the intermediate dose group, 2 mice died on day 14, 4 died on day 15, 1 mouse died on day 16, 2 died on day 19 and one mouse on day 21. The average number of days for this group was 16.50. In the lowest dose group, two mice died on days 12, 13, 14 and 15 and one died on days 16 and 17. The average number of days for this group was 14.1.

In the in vivo leukemia mouse model, P388, carbendazole increased the survival time of mice compared to the control group by 129% at 1000 mg / kg, 148% at 2000 mg / kg, and 189% at 4000 mg / kg.

Cancer model in mice

Carbendazim slowed tumor growth in a mouse, breast and lung cancer model. MXI breast cancers, inoculated subcutaneously under the skin of mice, were treated with a 500 mg / kg dose of carbendazim. Tumor growth slowed by 42%. Carbendazim slowed the growth of A549 lung tumor inoculated subcutaneously under the skin of mice by 57% at the same dose. Mass screening of mice with HT29 inoculated subcutaneous tumors under the skin of the mice slowed tumor growth by 54% at a dose of 2500 mg / kg carbendazim.

Cytoxan, which was administered to animals of the mouse model for breast cancer, significantly reduced tumor mass.

Carbendazim was administered to mice at doses of 4000, 5000, and 6000 mg / kg of body weight per foot. The tumor further decreased in size and its regrowth was limited even after 180 days of carbendazim treatment. Growth depended on the dose. The control group treated with cytoxan experienced repeated growth after 100 days, and after estrogen stimulation, rapid growth occurred after day 115. Even after estrogen stimulation, there was no significant change in tumor mass in the animals treated with carbendazim. After 130 days carbendazim (at 4000, 5000, and 6000 mg / kg body weight) was administered to mice previously treated with cytoxane. The tumor further decreased in size and its regrowth was limited even after 180 days.

In an in vivo leukemia (P388) mouse model, griseofulvin was shown to prolong survival compared to the control group by 156% at 4000 mg / kg, 188% at 5000 mg / kg, and 218% at 6000 mg / kg. kg.

Model of melanoma in mice

In the in vivo mouse model for melanoma, B16, carbendazim increased survival time compared to the control group by 131% at 1000 mg / kg, 163% at 2000 mg / kg, and 187% at 4000 mg / kg.

When carbendazim was combined with navelbine from 0.5 mg / kg to 2.0 mg / kg, the effective dose of carbendazim in the mouse model was lower in vivo.

Dose of carbendazim (Mg / kg) Dose of navelbine (Mg / kg) % increase in survival time compared to with the group untreated mice 4000 0.5 255 4000 1.0 298 4000 2.0 268 2000 0.5 259 2000 1.0 265 2000 2.0 287 1000 0, 5 207 1000 1.0 233 1000 2.0 245 - 0.5 190 - 1.0 245 - 2.0 265

In the in vivo mouse model for melanoma (B16), griseofulvin increased survival time compared to the untreated control group by 165% at 4000 mg / kg, by 179% at 5000 mg / kg and by 201% at 6000 mg / kg. Cytoxan 300 mg / kg prolonged survival by 192%.

Example 3

Evaluation of antiviral effect in human influenza virus

Female CD mice (from Charles River Breeding Labs, Portage, MI) were accepted for use at 5-7 days of age. The mice were 6 to 9 weeks of age and weighed approximately 20 to 28 g at the start of the test. All mice used in this study did not differ by age by more than 10 days. Mice were housed six times in bedding cages.

They were fed to the rodent diet 5002 (PMI, St. Luis Missouri) to taste.

Mice were exposed to human influenza virus, AT2 (Taiwan / 1/164). The organism was stored at -70 ° C. Prior to infection, the frozen vials were thawed and diluted to an appropriate concentration with buffered saline. Mice were anesthetized with halothane and a 50 μ 50 dose of virus was administered through the nose.

Test substances were administered at the concentrations and volume below. During Days 1 to 14, 10 mice from each group received test compounds by mouth wash. Saline control animals (10) received a comparable volume of saline compared to test compound mice. Dosing of the test substance was performed at 24 hour intervals. On day 0, about 4 hours after the second dose of test substance or saline, all mice were exposed to an infectious dose of virus that was calculated to ensure 90% mortality. Animals were observed daily for morbidity and mortality for 21 days after challenge.

test substance dose (mg / kg) mortality fluconazole 350 0 fluconazole 700 30 saline - 100 amantadine 75 0

At a dose of 175 mg / kg propiconazole, 40% of the mice survived compared to the saline control group where none survived. At a dose of 350 mg / kg propiconazole, 57% of the mice survived.

Example 4

Evaluation of antiviral effect in rhinovirus

In the WI-38 cell line rhinovirus type A-1 bulk study, propiconazole was effective at a dose of 32 gg / ml. A-36683 from Abbott, (S, S) -1,2-bis (5-methoxy-2-benzimidazolyl) -1,2-ethanediol, was a positive control. For group A-36683, the therapeutic index was 1000 to 3200. Propiconazole had a therapeutic index of 1 to 3 (see Schleicher et al., Applied Microbiology, 23, No. 1, 113-116 (1972)).

In a bulk study for W1-38 cell line A-1 rhinovirus, griseofulvin was effective at a dose of 100 pg / ml. A-36683 from Abbott, (S, S) -1,2-bis (5-methoxy-2-benzimidazolyl) -1,2-ethanediol, was a positive control. For group A-36683, the therapeutic index was 1000 to 3200. Griseofulvin had a therapeutic index of 1 to 2 (see Schleicher et al., Applied Microbiology, 23, No. 1, 113-116 (1972)).

Example 5

Human tumor colony formation assay in vitro

Solid tumors that were removed by the patients were crushed into 2-5 mm fragments and immediately placed in Mc Coy medium 5A with 10%, heat-inactivated newborn calf serum and 1% penicillin / streptomycin. Within 4 hours, these solid tumors were mechanically divided by means of scissors, passed through a stainless steel sieve no. 100, via needle no. 25 and washed with McCoy medium as described above. Intra-abdominal, pleural and pericardial fluid and bone marrow were obtained by standard procedures. The fluids or marrow were placed in sterile containers containing 10 units of preservative-free heparin per ml of malignant fluid or pulp. After centrifugation for 10 minutes at 150G, cells were removed and washed with McCoy medium with 10% heat-inactivated calf serum. The viability of the cell suspension was determined by trypan blue hemocytometer.

Cells were cloned and suspended in 0.3% agar, supplemented with CMRL 1066, supplemented with 15%, heat inactivated horse serum, penicillin (100 units / ml), streptomycin (2 mg / ml), glutamine (2 mmol), insulin (3). units / ml), asparagine (0.6 mg / ml) and HEPES buffer (2 mmol). For a continuous exposure test, each substance is added to the higher mixture. Cells are placed in 35 mm Petri dishes between two agar layers to prevent fibroblast growth. Three plates are prepared for each experimental point. Plates are placed in a 37 ° C incubator and removed on day 14, counting for each number of colonies on each plate. The number of colonies (defined as 50 cells) formed on the three drug-treated plates is compared to the number of colonies formed on the three control plates. The percentage of colonies that survive at a certain concentration of the substance is determined. Three plates as positive controls are used to determine the degree of survival. Sodium orthovanadate at a concentration of 200 µg / ml is used as a positive control. The assay is evaluated if less than 30% of colonies are present on the positive control plates compared to the untreated control plates.

In a single dose experiment, propiconazole at concentrations of 0.5 and 5.0 pg / ml was inactive (0/1) against tumors. At a concentration of 50.0 gg / ml in the continuous exposure experiment, propiconazole was effective against colon, lung (non-small cell), melanoma, and ovarian cancer. 6 out of 8 were <50% survival.

In a single dose experiment, griseofulvin at concentrations of 0.5 and 5.0 gg / ml was inactive (0/1) against tumors. At a concentration of 50.0 gg / ml in a continuous exposure experiment, griseofulvin was effective against colon, lung (non-small cell), melanoma, and ovarian cancer. 5 out of 6 were <50% survival.

In a single dose experiment, fluconazole at concentrations of 0.5 and 5.0 gg / ml was ineffective (0/3 and 0/1, 13) against tumors. At a concentration of 50.0 pg / ml in a continuous exposure experiment, fluconazole was effective against non-small cell lung tumors and in particular ovarian cancer. In 4 of 13, <50% was survival.

f * 0 «ΜΜ-Α4

Claims (34)

A pharmaceutical composition for the treatment of viral infections in a warm-blooded mammal, comprising from 700 mg to 6000 mg of a substance selected from the group consisting of herbicides, fungicides, herbicide derivatives, fungicide derivatives and mixtures thereof with a pharmaceutically acceptable carrier. Pharmaceutical preparation according to claim 1, characterized in that the viral infection is caused by a virus which contains genetic material from plants, fungi and fungi. Pharmaceutical preparation according to claim 1 or 2, characterized in that the viral infection is an HIV infection. Pharmaceutical preparation according to claim 3, characterized in that an antiviral agent, preferably a transcriptase inhibitor or a protease inhibitor, is added to the compound. Pharmaceutical preparation according to claim 1, 2, 3 or 4, characterized in that it contains 3000 mg to 6000 mg of said herbicide or fungicide. Pharmaceutical preparation according to claim 5, characterized in that it is prepared in a liquid form selected from the group consisting of aqueous solutions, alcohol solutions, emulsions, suspensions and suspensions formed from non-effervescent and effervescent formulations and suspensions in pharmaceutically acceptable fats or oils. A pharmaceutical composition according to claim 3, 4 or 5, wherein the HIV medicament is added to said pharmaceutical composition. Pharmaceutical preparation according to claim 1, 2, 3, 4, 5, 6 or 7, characterized in that a potentiator is added to said pharmaceutical preparation. A pharmaceutical composition for the treatment of malignant, comprising from 700 mg to 6000 mg of a substance selected from the group consisting of herbicides, fungicides, herbicide derivatives, fungicide derivatives, or a pharmaceutically acceptable salt of an organic or inorganic acid with a pharmaceutically acceptable carrier. Pharmaceutical preparation according to claim 9, characterized in that the tumor cells comprise genetic material from plants, fungi and fungi. Pharmaceutical preparation according to claim 9 or 10, characterized in that a potentiator is added to said pharmaceutical preparation. Pharmaceutical preparation according to claim 10 or 11, characterized in that it contains 3000 mg to 6000 mg of said herbicide or fungicide or fungicide derivative or herbicide derivative. 13. The pharmaceutical composition of claim 12, wherein a chemotherapeutic agent is added to said pharmaceutical composition. Pharmaceutical preparation according to claim 13, characterized in that said chemotherapeutic agent is selected from the group consisting of DNA agents, antimetabolites or tubulin agents, preferably asparaginase, hydroxyurea, cisplatin, cyclophosphamide, altretamine, bleomycin, dactinomycin, doxorubicin, etoposide, teniposide and plicamycin, methotrexate, fluorouracil, fluorodeoxyuridine, CB3717, azacytidine, cytarabine, floxuridine, mercaptopurine, 6-thiogvanine, pentostatin, cyctrabine and fludarabine. 15. The pharmaceutical composition of claim 14, wherein said chemotherapeutic agent is in the form of liposomal particles. Pharmaceutical preparation according to claim 9, 10, 11, 12, 13, 14 or 15, characterized in that 0.5 mg to 400 mg of the chemotherapeutic agent is used. Pharmaceutical preparation according to claim 13, 14, 15 or 16, characterized in that said herbicide or fungicide is in liquid form selected from the group consisting of aqueous solutions, alcoholic solutions, emulsions, suspensions and suspensions formed from non-effervescent and effervescent formulations and suspensions in pharmaceutically acceptable fats or oils. 18. A method for the preparation of a pharmaceutical composition for the treatment of viral infections in a warm-blooded mammal, comprising mixing a safe and effective amount of a substance selected from the group consisting of herbicides, fungicides, herbicide derivatives, fungicide derivatives, pharmaceutically acceptable salts thereof. and mixtures thereof with a pharmaceutically acceptable carrier. 19. The method according to claim 1, wherein said substance is prepared for the treatment of a viral infection caused by a virus which comprises genetic material from plants, fungi and fungi. 20. The method of claim 1 or 2, wherein said viral infection is an HIV infection. A method according to claim 3, characterized in that an antiviral agent, preferably a transcriptase inhibitor or a protease inhibitor, is added to the compound. A process according to claim 1, 2, 3 or 4, characterized in that 3000 mg to 6000 mg of said herbicide or fungicide is used for its preparation. The method of claim 5, wherein the mixture is prepared in a liquid form selected from the group consisting of aqueous solutions, alcoholic solutions, emulsions, suspensions and suspensions formed from non-effervescent and effervescent formulations and suspensions in pharmaceutically acceptable fats or oils. . A method according to claim 3, 4 or 5, characterized in that an HIV medicament is added to said herbicide or fungicide. 25. A method according to claim 1, 2, 3, 4, 5, 6 or 7, characterized in that a potentiator is added to said pharmaceutical composition. 26. A method of preparing a pharmaceutical for the treatment of malignant tumors, comprising adding a safe and effective amount of a herbicide, fungicide, herbicide derivative, fungicide derivative, a pharmaceutically acceptable salt of an organic or inorganic acid thereof with a pharmaceutically acceptable carrier. 27. The method of claim 9 wherein said pharmaceutical composition is prepared for the treatment of a malignant tumor comprising genetic material from plants, fungi, and fungi. The method of claim 9 or 10, wherein a potentiator is added to said pharmaceutical composition. A process according to claim 11, wherein 2 mg to 4000 mg of said herbicide or fungicide or fungicide derivative or herbicide derivative are added. The method of claim 11, wherein a chemotherapeutic agent is added to said herbicide or fungicide. 31. The method of claim 13, wherein said chemotherapeutic agent is selected from the group consisting of DNA agents, antimetabolites, or tubulin agents, preferably asparaginase, hydroxyurea, cisplatin, cyclophosphamide, altretamine, bleomycin, dactinomycin. , doxorubicin, etoposide, teniposide and plicamycin, methotrexate, fluorouracil, fluorodeoxyuridine, CB3717, azacytidine, cytarabine, floxuridine, mercaptopurine, 6-thiogvanine, pentostatin, cyctrabine and fludarabine. 32. The method of claim 14 wherein said chemotherapeutic agent is in the form of liposomal particles. Process according to claim 9, 10, 11, 12, 13, 14 or 15, characterized in that 150 mg to 4000 mg of said herbicide or fungicide are added and 0.5 mg to 400 mg of the chemotherapeutic agent are used. I I 34. The method of claim 13, 14, 15 or 16 wherein said herbicide or fungicide is prepared in a liquid form selected from the group consisting of aqueous solutions, alcoholic solutions, emulsions, suspensions, and suspensions formed from non-effervescent and effervescent products. preparations and suspensions in pharmaceutically acceptable fats or oils.