COMPOUNDS AND METHODS OF USING THEM IN THE TREATMENT OF CANCER OR VIRAL INFECTIONSTECHNICAL FIELD The present invention relates to benzimidazole derivatives and their use for the treatment of cancer or a viral infection in warm-blooded animals, particularly in humans and other mammals. The methods can use this compound in combination with an enhancer or a chemotherapeutic agent.
BACKGROUND OF THE INVENTION Cancers are a leading cause of death in animals and humans. The exact cause of the cancer is unknown, although several researchers have shown links between certain activities such as smoking or exposure to carcinogens and the incidence of certain types of cancers and tumors. Many types of chemotherapeutic agents have been shown to be effective against cancers and tumor cells, although not all types of cancers and tumors respond to these agents. Unfortunately, many of these agents also destroy normal cells. The exact mechanism for the action of these chemotherapeutic agents is not always known. ? Despite advances in the field of cancer treatment, the main therapies to date are surgery, radiation and chemotherapy. The chemotherapeutic proposals are for the fight against cancers that spread or those that are particularly aggressive. These cytocidal or cytostatic agents work best on cancers with large growth factors, that is, those whose cells are dividing rapidly. ? the date, the hormones, in particular estrogen, progesterone, and threotide, and some antibiotics produced by a variety of microbes, alkylating agents, and anti-metabolites form the major part of the therapies available to oncologists. Ideally, cytotoxic agents having specificity for cancer and tumor cells, while not affecting normal cells, could be conveniently desired. Unfortunately, none have been found and instead agents have been used that focus especially on rapidly dividing cells (tumor and normal). Clearly, the development of materials that focus on cancer cells due to some unique specificity for them could be an important advance. Alternatively, materials that were cytotoxic to cancer cells while they exert mild effects on normal cells would be desirable. The Human Immunodeficiency Virus (HIV), the etiological agent for AIDS (acquired immunodeficiency syndrome), is a member of lentivirus, a subfamily of retroviruses. HIV integrates its genetic information into the host's genome. More importantly, HIV infects and invades cells of the immune system; It breaks the body's immune system and makes the patient susceptible to opportunistic infections and neoplasms. HIV-1 is cytopathic for T4 lymphocytes, the cells of the immune system that express the antigen for CD4 surface cell differentiation. In addition to CD4 + T lymphocytes, the HIV host boundary includes cells of the mononuclear phagocytic lineage, including blood monocytes, tissue macrophages, skin Langerhans cells, and dendritic reticulum cells within the lymph nodes.
The precursor cells in the bone marrow are released into the blood in an immature circulating form known as monocytes. Monocytes use blood strictly as a means of transport. Once they reach the place where they will be used, they leave the blood and end the differentiation in the macrophages. Cells from the monkey citrus / macrophage lineage are a major target population for HIV infection in the body and are thought to provide deposits of virus to spread the infection throughout the body. HIV is also a neurotropic, capable of infecting monocytes and macrophages in the central nervous system causing severe neurological damage. They can interact and fuse with CD4-carrier T lymphocytes, causing depletion of the T lymphocyte and thus contributing to the pathogenesis of AIDS. The evolution of HIV infection to AIDS is mainly determined by the effects of HIV on the cells it infects, including CD4 + T lymphocytes and macrophages. In turn, cell activation, differentiation and proliferation regulate HIV infection and duplication in these cells. HIV and other lentiviruses can proliferate in terminally differentiated, non-proliferating macrophages and T lymphocytes with arrested growth. It is believed that this ability of lentiviruses, including HIV, to reproduce in non-proliferating cells, particularly in macrophages, is unique among ret roviruses. Due to the problems mentioned above in the art, the inventors of the present have sought improvements and provide these improvements herein. Carbendazim, or 2-methoxy-carbonylaminobenzimidazole, has been studied as a treatment for cancer. See US 5, 767, 138 issued June 16, 1998 to J. B. Camden. Carbendazim is metabolized in the body through the hydroxylation of the benzene ring, p incipiently in position 5. The metabolite is not as active in the treatment of cancer as 2-methoxycarbonylaminobenzimidazole. In addition, this compound is not very soluble. A derivative that maintains the activity of the benzimidazole even though it is more soluble is more preferred. It has been found that substituted benzimidazole carbamates, and in particular, those claimed herein are more soluble, and even maintain their cytotoxic behavior.
BRIEF DESCRIPTION OF THE INVENTION The compounds of the present invention are represented by the following formula A:wherein, R is hydrogen, -COOR2 or -CONHR2; Ri is hydrogen, -COOR3 or -CONHR3; each R2 and R3 is independently alkyl, haloalkyl, alkenyl, haloalkenyl, cycloalkyl, cycloalkyl, heterocycloalkyl, heterocycloalkyl, substituted or unsubstituted benzyl, hydroxyalkyl, alkoxyalkyl, poly (alkoxy) alkyl, hydroxyalkoxyalkyl, hydroxypoly (alkoxy) alkyl, haloalkoxyalkyl, halopoli (alkoxy) alkyl, or aminoalkyl; each X and Y is independently hydrogen, alkyl, alkenyl, cycloalkyl, haloalkyl, haloalkenyl, bromo, chloro, fluoro, nitro, or amino; and n is 1, 2 or 3.
The benzyl group may optionally be substituted with one or more substitutes is nitro, carboxy, hydroxy, alkyl, alkoxy, or halide. Also included in the present invention are the pharmaceutically acceptable salts of the benzimidazole derivatives of the formula A. The prodrugs of the compounds of the formula? Are also included in the invention. In a presently preferred embodiment of the invention, the benzimidazole derivatives of the invention are of the formula A-1 or A-2:A-1 A2and more preferably the compounds are of the formula A-1 or A-2 where each X and Y is hydrogen. In another currently preferred embodiment of the invention, the benzimidazole derivatives are of the formula A-3:, A-3and most preferably, the compounds are of the formula A-3, wherein X is hydrogen and Y is hydrogen or halogen; the halogen is preferably chloro. In another currently preferred embodiment of the invention, the benzimidazole derivatives are of the formula A-4:A-4and most preferably, the compounds are of the formula A-4, wherein X is hydrogen and Y is hydrogen or halogen; the halogen is preferably chlorine. By the present invention methods for the treatment of warm-blooded animals, and in particular, humans and other mammals that are affected by cancer or viral infection, are provided, the methods comprising administering to the animal a therapeutically effective amount of a derivative of benzimidazole of the formula A, or a salt or a prodrug thereof.
DETAILED DESCRIPTION OF THE INVENTION A. Definitions The term "alkyl" refers to a saturated monovalent hydrocarbon radical completely of 1 to 12 carbon atoms. It can be straight or branched chain. Alkyl groups containing from 1 to 10 carbon atoms, with 2 to 8 carbon atoms which are particularly preferred, are preferred. The term "alkenyl" refers to an unsaturated monovalent hydrocarbon radical of 2 to 12 carbon atoms containing only carbon and hydrogen and having one or more double bonds. It can be straight or branched chain. Alkenyl groups containing from 2 to 10 carbon atoms, with 2 to 8 carbon atoms which are particularly preferred, are preferred. The term "alkoxy" means the group -OR 'wherein R' is alkyl. Alkoxy groups having 1 to 10 carbon atoms, more preferably 2 to 8 carbon atoms, are preferred.
The term "α 1 -alcoxy" refers to an alkoxy group covalently bonded to an alkyl group. The alkoxy group contains from 1 to 12, preferably from 1 to 6, carbon atoms. The alkoxy group can be substituted with one or more hydroxyl groups (a "hydroxyalkoxyalkyl") or with one or more halogen atoms (a "haloalkoxyalkyl"); preferably hydroxyl or the halogen is at the terminal end of the alkoxyalkyl substituent. The term "poly (alkoxy) alkyl" represents from 2 to 200, preferably from 2 to 20, alkoxy groups covalently bonded in a straight or branched configuration and attached to an alkyl group. The linear pol i (al coxi) entities have a structure such as - (CH2) m -0- (CH2) m -0- (CH2) m -0- (CH2) m- ... -0-CmH2m + 1, where "m" is a whole number, same or different along the length of the chain. Branched entities have two or more groups (-0- (CHt) m-) linked to a third common group (-0- (CHt) m-), where "t" has a value that is selected independently of 0, 1 and 2 for each group (CHt) m. Linear configurations are preferred. The number of repeating groups (-O- (CHt) m-) within a substituent can be up to 200, preferably from 2 to 20, more preferably from 2 to 7, and most preferably 2-5. . The individual alkoxy groups may be the same or different, and the individual alkoxy groups preferably contain from 1 to 6 carbon atoms each, and most preferably from 1 to 3 carbon atoms each. A currently preferred poly (alkoxy) alkyl is - (CH2) y- (OCH2CH2) X-OCH3 or - (CH2) y- (OCH2CH2) x-OCH2CH3, where y = 1-4 and x = 1-100, preferably 1-10, and more preferably 2-5. The individual alkoxy groups can be substituted with one or more hydroxyl groups (a "hydroxypoly (alkoxy) alkyl") or with one or more halogen atoms (a "halo (alkoxy (alkyl)"); preferably the hydroxyl or halogen is at the terminal end of the poly (alkoxy) alkyl substituent. "Heterocycle" designates a heterocyclic group; that is, a closed ring structure, usually 5 or 6 members, in which one or more of the atoms in the ring is a different element of the carbon atom, such as for example sulfur, nitrogen, or oxygen. A heterocyclic group may be, but is not limited to, pyridine, pyrrole, furan, thiophene, morpholine and purine, optionally substituted with one or more substituents nitro, carboxy, suifonic acid, hydroxy, alkyl, alkoxy, or halide. The term "amino" refers to primary amines (-NH2), to secondary amines (-NHR '), and tertiary amines (-NR'R "), wherein R' and R" are the same or different substituent groups, such as alkyl, alkenyl, halogen, hydroxy, and the like. "Independently" means that two or more groups immediately preceding the term are identical or different; that is, the selection of one of the following list, the term does not affect the selection of the other or others. "Substituted" encompasses both individual and multiple substitutions, the latter include multiple substitutions by the same substituent, as well as mixtures of different substituents. The term "optional" or "optionally" means that the event or circumstance described below may or may not be presented, and that the description includes the cases in which the event or circumstance is presented and the cases in which it does not. For example, "optionally substituted phenyl" means that the phenyl may or may not be substituted and that the description includes both unsubstituted phenyl and phenyl where there is a substitution. As another example, "optionally" followed by "converting the free base to that of acid addition" means that this conversion may or may not be carried out in order that the process described is within the invention, and the invention includes those processes where the free base is converted to the acid addition salt and those processes in which it does not. As used herein, "therapeutically effective amount" means the concentration or amount or level of the compound of the present invention that can achieve a particular medical purpose, such as the control or destruction of cancer cells, virally infected cells, or virus without producing unacceptable toxic symptoms. The term "safe and effective amount" refers to the amount of a component that is sufficient to provide a desired therapeutic response without undue adverse side effects (such as toxicity, irritation, or allergic response) consistent with a reasonable proportion of bene fi t. ri is go when used in the manner of this invention. The specific "safe and effective amount" will vary with these factors depending on the particular condition being treated, the physical condition of the patient, the type of mammal being treated, the duration of treatment, the nature of the concurrent therapy (if there is), and the specific formulations used and the structure of the compounds or their salts. As used herein, a "pharmaceutical addition salt" or "pharmaceutically acceptable salt" is a salt of a benzimidazole-derivative compound with an organic or inorganic acid. These preferred acid addition salts are chlorides, bromides, sulfates, nitrates, phosphates, sulfonates, formates, tartrates, maleates, maleates, citrates, benzoates, salicylates, ascorbates, and others known to those of ordinary skill in the art. As used herein, the term "prodrug" refers to a form of a benzimidazole-derived compound that has minimal therapeutic activity until it is converted to its desired biologically active form. A prodrug is a compound having one or more functional groups or carriers covalently attached thereto, the functional or carrier groups are removed from the compound by metabolic processes within the body to form the respective bioactive compound. As used herein, the term "metabolite" refers to the decomposition or final product of a compound derived from benzimidazole or its salt produced by metabolism or biotransformation in the animal or human body.; for example, biotransformation to a more polar molecule such as, for example, by oxidation, reduction, or hydrolysis, or to a conjugate (see Goodman and Gilman, "The Pharmacological Basis of Therapeutics" 8th Ed., Pergamon Press, Gilman et al. (eds.), 1990 for an analysis of biotrans formation.] As used herein, the metabolite of a benzimidazole derivative compound or its salt may be the biologically active form of the compound in the body. An analysis for the activity of a metabolite of a benzimidazole derivative of the present invention is known to one of skill in the art in light of the present disclosure, for example, the efficacy test against a virus in vitro or in vivo. In the sense in which it is used herein, a "subject in need thereof" is a warm-blooded animal having cancer or having a viral infection.In the sense in which it is used herein, "cancer" is ref iere to all types of cancers, or neoplasms or benign or malignant tumors. In one embodiment, those cancers that attack healthy normal blood cells or bone marrow are contemplated by the present invention. Preferred cancers for treatment using the methods provided herein include carcinoma, sarcoma, lymphoma, or leukemia. By "carcinoma" is meant a benign or malignant epithelial tumor and includes, but is not limited to: breast carcinoma, prostate carcinoma, non-small cell lung carcinoma, colon carcinoma, CNS carcinoma, melanoma carcinoma, ovarian carcinoma, or renal carcinoma. A preferred host is a human host. A "viral infection", in the sense in which it is used herein, means an infection due to a DNA virus or an RNA virus (retrovirus). Examples of a double-stranded DNA virus are the herpes virus and the influenza virus. The human immunodeficiency virus (HIV) is a prototype for retroviruses, that is, viruses that use reverse transcription to reproduce. However, certain DNA viruses use, in part, reverse transcription mechanisms to reproduce, such as, for example, hepatitis B virus. "Viruses" include retroviruses such as, for example, HIV or HTLV, influenza, rhinovirus. , herpes, hepatitis, or the like. As used herein, a benzimidazole derivative of the formulas A and A-1 through A-4, or a pharmaceutical salt thereof or a prodrug thereof, are the "compounds of the present invention". These compounds are further shown at point B below. As used herein, the term "chemotherapeutic agents" includes agents interactive with DNA, antimetabolites, tubulin-interactive agents, hormonal agents and others, such as asparaginase or hydroxyurea, and are additionally shown in point D above. ahead. In the sense in which it is used herein, "enhancers" are materials that affect the inrun system or enhance the effectiveness of a compound of the present invention and are further shown at point E herein. Following the old patent law convention, the terms "one" and "ones" mean "one or more" when used in this application, including claims.
B. Benzimidazole Derivatives The benzimidazole derivatives of the present invention are those of the formula A, as shown above. In one embodiment of the invention, the presently preferred compounds are those of the formulas A-1 and A-2. In another embodiment, the presently preferred compounds are those of the formulas A-3 and A-4. The pharmaceutically acceptable salts of the benzimidazole compounds are considered to be within the scope of the compounds of the present invention. They are salts with an organic or inorganic acid. Preferred acid addition salts are chlorides, bromides, sulfates, nitrates, phosphates, sulfonates, formates, tartrates, maleates, maleates, citrates, benzoates, salicylates, ascorbates, or the like. These salts can be synthesized from the compound of the present invention, or the derivative thereof, which contains a basic or acid entity, by conventional chemical methods. In general, these salts can be prepared by reacting a free acid or a basic form of the compound, or derivative thereof, with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two. In general, nonaqueous media similar to ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. Additional suitable salts can be found in Remington: The Science and Practice of Pharmacy, 19a. ed. , Mack Publishing Company, Easton, PA. , 1995, p. 1457. Pharmaceutically acceptable salts of the compounds of the present invention include the conventional non-toxic salts or the quaternary ammonium salt of the compounds or derivatives formed, for example, from non-toxic inorganic or organic acids. For example, these conventional non-toxic salts include those derived from inorganic acids, such as, for example, hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric, or the like; and salts prepared from organic acids, such as, for example, acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, maleic, hydroxylaleic, f-enylacetic, glutamic, benzoic, salicylic, sulfanilic, 2- acet oxybenzoic, fumaric, toluenesulfonic, ulonic, ethanedisulfonic, oxalic, isethionic, or the like. Preferred acid addition salts are chlorides, bromides, sulfates, nitrates, phosphates, sulfonates, formates, tartrates, maleates, maleates, citrates, benzoates, salicylates, ascorbates, or the like. A presently preferred salt is the hydrochloride salt. Also included within the scope of the compound, or the salts thereof, useful for the present invention are the prodrugs thereof. As used herein, a "prodrug" is a drug covalently linked to a carrier wherein the release of the drug occurs in vivo when the prodrug is administered to a mammalian subject. The prodrugs of the compounds of the present invention are prepared by modifying the functional groups present in the compounds in such a way that the modifications are split, either in routine manipulation or in vivo, to provide the desired compound. Prodrugs include compounds wherein the hydroxy, amine, or sulfhydryl groups bind to any group that, when administered to a mammalian subject, decomposes to form a free hydroxyl, amino, or sulfhydryl group, respectively. Examples of prodrugs include, but are not limited to: acetate, formate, or alcohol benzoate derivatives or functional amine groups in the compounds of the present invention; phosphate esters, dimethylglycine esters, aminoalkylbenzyl esters, aminoalkyl esters or carboxyalkyl esters of alcohol or phenol functional groups in the compounds of the present invention; or the like.
Parameters of the synthetic reaction The terms "solvent""inert organic solvent" and "inert solvent" mean a solvent that is passive or non-reactive under the reaction conditions that are being described together with them (including, for example, benzene, toluene, acetonitrile, tetrahydrofuran ("THF") ), dimethylformamide ("DMF"), chloroform, methylene chloride (or dichloromethane), diethyl ether, methanol, pyridine, and the like). Unless otherwise specified, the solvents used in the reactions of the present invention are inert organic solvents.
The temperature of the reaction can vary widely depending on the reactivity of the reactants. However, the temperature should not be so high that it decomposes the reagents or so low that it causes the inhibition of condensation or freezing of the solvent. ? Unless otherwise specified, the reactions described herein are carried out at atmospheric pressure over a temperature range from dry ice temperature to about 100 ° C, more preferably between about 10 ° C and 50 ° C, and most preferably at "ambient" or "ambient" ("RT") temperature, for example, about 20 ° C. ? Unless otherwise specified, the reaction times and conditions are intended to be approximate. The time required in the present for the reactions will depend to a large extent on the temperature that will be used and the relative reactivities of the starting materials. Therefore, the reaction time can vary widely, for example between about five minutes and two days. Various known techniques can be used such as, for example, different types of chromatography, especially thin layer chromatography ("TLC"), gas chromatography ("GC"), or spectroscopy to monitor the progress of the reaction by the disappearance of the starting compound or compounds. The isolation and purification of the compounds and intermediates described herein can be effected, if desired, by any convenient separation or purification process such as, for example, filtration, extraction, crystallization, column chromatography, thin layer chromatography. , thick layer chromatography, centrifugal chromatography, or preparative HPLC, or a combination of these procedures. Specific illustrations of convenient separation and isolation procedures can be found by reference to the examples below. However, of course other equivalent separation or isolation procedures can also be used.
Synthesis of the compounds of Formula A The compounds of the invention are prepared by modifying the benzimidazole nucleus:In a synthetic process, the commercially available 2-aminobenzimidazole (101) can be reacted with a variety of chloroformates or isocyanates in pyridine to form, respectively, the carbamates (105) or ureas (103) in the N3 position. In this manner, benzimidazole derivatives substituted at the N3 position of the formula A-1 (105) or A-2 (103) with a free 2-amino group are synthesized (Reaction Scheme 1).
Reaction Scheme 1:Upon heating the N3-acyl analogs (103, 105) in pyridine, the acyl groups can be induced to migrate to the 2-amino position to provide the benzimidazole derivatives of the formula A-3 or A-4 (205 and 203, respectively) (Reaction Scheme 2).203 205Reaction Scheme 2: When it is desired to synthesize a compound of the formula A having a chlorine atom in the 5 (6) position, the 4-chloro-1,2-phenylenediamine 301 is condensed with cyanogen bromide (Zou, R . et al., J. Med. Chem. 40: 811-818 (1997)) to provide the corresponding chloro 2-aminobenzimidazole 303. The benzimidazole 303 can then be heated with an isocyanate in pyridine to form the chloro analogues 2-aminourea-305 (Reaction Scheme 3).
Reaction Scheme 3Another method of preparing the compounds of formula A-3 or A-4, the commercially available methyl 2-aminobenzimidazole carbamate fungicide (carbenda zim, 401), is treated with a suitable alcohol or amine with or without added toluene as the solvent in presence of the aluminum isopropoxide catalyst to provide the new carbamates (403) and the ureas (405), respectively (Reaction Scheme 4).
Reaction Scheme:405Accepted preferred embodiments In one embodiment of the present invention, the currently preferred benzimidazole derivatives are those where Rx and X are hydrogen, and Y is hydrogen or chlorine and is in position 5 (6). Also preferred are those compounds where Rx, X and Y are hydrogen and R is selected from those groups listed in Table 1: The solubility is based on a normal measurement used in medicinal chemistry, the octanol-water partition coefficient, LogP, Lower Log) values indicate superior aqueous solubility. There are a variety of methods to estimate this value by a proposed structure using computer calculations. The low LogP solubility values were determined using the atomic typing method of Ghose, Pritchett et al. (J. Cornput, Chem 9 (l): 80-90, 1988). Carbendazim has a LogP of 1,302.
TABLE 1 Comp. Do not . R LogP 1-1 -COOCH2CH = CH2 2.408 1-2 -COOCH2CH2CH3 3.202 1-3 -C0NHCH2C¾C1 1.721 1-4 -C0NHCH2Ph 2.794 1-5 -COOCH2Ph 3.44 1-6 -CONHCH2CH2CH3 1.829 1-7 -COOCH, CH2CH3 2.479 1- 8 -COOCH2CH2CH2N (CH3) 2-1-9 -CONHCH2Ph (-2-OH) -1- 10 -COOCH2CH2-morpholino-1-11 -COOCH (CH3) (cyclopropyl) - In another embodiment of the invention, the derivatives of Currently preferred benzimidazole are those where R is hydrogen, X is hydrogen, and Y is hydrogen or chlorine and is in the 5 (6) position. Also preferred are those compounds where R and X are hydrogen, Y is hydrogen or chlorine at the 5 (6) position, and Ri is selected from those groups listed in Table 2:TABLE 2 Comp. Do not . Ri LogP 2-1 -C0NHCH2CH2N (CH3) 2 0.531 2-2 -COOCH2CH = CH2 2.042 2-3 -COO (CH2CH20) 2CH2CH3 1.315 2-4 -CONHCH2CH2OCH2CH2OH 0.044 2-5 -COOCH2CH2OCH2CH2CI 1.841 2-6 -COOCH2CH2Cl 2.005 2- 7 -COOCH2CH2OCH2CH2OH 0.694 2-8 -COOCH2CH2CH3 2.113 2-9 -COOC¾Ph 3.078 2-10 -C0NHCHZCH2C1 1.355 2-10 * -C0NHCH2CH2C1 1.873 2-11 -CONHCH2Ph 2.428 2-12 -CONHCH2CH2CH3 1463 Comp. Do not . Ri LogP 2-12 * -CONHCH2CH2CH3 1.981 2-13 -CONHCH3-2-13 * -CONHCH3 1.170 2-14 -CONHCH2Ph (-2-OH) -2-15 -COOCH2CH2-morpholino-2-16 -COOCH (CH3) (cyclopropyl) - * Y = Cl, in position 5 (6)C. Selection Analysis Selection analyzes to determine those cancers susceptible to a treatment using the compounds of the present invention include incubation cell line models representing specific cancers as established, for example, by the National Cancer Institute, in the presence of and absence of these compounds. Viability of the cells can be determined by MTT analysis (Promega S.A., Madison, WI 53711), or SRB analysis (sulforhodamine B) (Skehan, et al., JNCI, 82:13, 1107, 1990). Susceptibility to these compounds exists when viability in the presence of a compound of the present invention has less than that viability in the absence of that compound.
Exemplary cell line models that represent specific cancers include, but are not limited to, the following: Non-small cell lung cancer: NCIH23, NCIH324, NCIH522, A549 / ATCC, A549 (ASC), CALU1, EKVX, NCIH125M, NCIH226, NCIH520, SKMES1, NCIH322M, NCIH358M, NCIH460, NCIH292, HOP62, HOP18, HOP19, HOP92, LXFL 529, S 1573, LXFS 650L, ML1019, ML1076, ML1045, or UABLG22; Small cell lung cancer: NCIH69,NCIH146, NCIH82, NCIH524, DMS 114, D S 273, HOP27, SHP77, or RHOS; Colon cancer: HT29, HCC2998, HCT116, LOVO, SW1116, SW620, COLO 205, DLD1, IDR, COLO 320DM, HCT15, CXF 280, M12, KM20L2, COLO 741, CXF 264L, COLO 746, UABC02, MLI059, CAC02, HT29 / PAR, HT29 / DR1, or NB4; Breast cancer: CF7, MCF7 / ADRRES, ZR751, ZR7530, MDAMB231 / A CC, HS 578T, UISCBCA1, MCF7 / ATCC, SKBR3, MDAMB 435, MDAN, BT549, T47D, MDAMB231, AXF 401, B 474, or MDAMB 468; Ovarian cancer: OVCAR3, OVCAR4, OVCAR5, OVCAR8, A2780, IGROV1, SKOV3, OVXF 899, A1336, or ES2;Leukemia: P388, P3888 / ADR, CCRFCEM, CCRFSB, K562, M0LT4, L1210, HL60 (TB), RPMI8226, SR, or K562 / ADR; Fibroblast: IMR90, or CCD19LU; Kidney cancer: U031, SN12C, SN12S1, SN12K1,SN12L1, SN12A1, A498, A704, CAKI1, RXF 393, RXF631, 7860, SW156, TK164, 769P, SS78, ACHN, TK10, RXF 486L,, UOK57, or UOK57LN; Melanoma: LOX I VI, MAL E3, RPMI7951, SKMEL2, SKMEL5, SKMEL28, SKMEL31, UCSD 242L, UCSD 354L, M14, M19MEL, UACC62, UACC257, MEXF 514L, or UABMEL3; Prostate cancer: PC 3, PC3M, DU145, LNCAP, 1013L, UMSCP1, IS, JE, RER, MRM, DHM, AG, RB, RVP, FC, WAE, DB / SMC, JCAl, ND1, WMF, TSUPRI, JECA , GDP, TIO, WB, RVP1, or WLL; CNS cancer: SNB7, SNB19, SNB44, SNB56, SNB75, SNB78, U251, TE671, SF268, SF295, SF539, XF 498, S 1088, SW 1783, U87 MG, SF767, SF763, A172, or SMSKCNY; . Bone / muscle: A204 / ATCC, OHS, TE85, A673, CHA59, MHM 25, RH18, RH30, or RD; and Lymphoma: AS283, HT, KD488, PA682, SUDHL7, RL, DB, SUDHL1, S U DHL 4, SUDHL10, NÜDUL1, or HUT 102.
D. Chemotherapeutic agents Chemotherapeutic agents in general are grouped as interactive agents with DNA, antimetabolites, tubulin-interactive agents, hormonal agents, other agents such as, for example, asparaginase or hydroxyurea, and the agents as set forth in Table 3. Each of the groups of chemotherapeutic agents can be further divided by the type of agent. activity or compound. The chemotherapeutic agents used in combination with a compound of the present invention, or the salts thereof of the present invention can be selected from any of these groups but are not limited thereto. For a detailed analysis of chemotherapeutic agents and their methods of administration, see Dorr, et al., Cancer Chemotherapy Handbook, 2a. edition, pages 15-34, Appleton &; Lange (Connecticut, 1994) incorporated herein by reference. DNA-interacting agents include alkylating agents, for example, cisplatin, cyclophosphamide, alt retamine; agents for DNA strand breakage, such as, for example, bleomycin; topoisomerase II intercalating inhibitors, (eg, dactinomycin and doxorubicin); non-intercalating inhibitors of topoisomerase II such as, for example, etoposide and teniposide; and minor channel binder plicamidine with DNA, for example. Alkylating agents form covalent chemical adducts with DNA, cellular RNA or protein molecules, or with minor amino acids, glutathione, or similar chemicals. In general, alkylating agents react with a nucleophilic atom on a cellular constituent, such as for example, an amino, carboxyl, phosphate, or sulfhydryl group on nucleic acids, proteins, amino acids, or on glutathione. The mechanism and function of these alkylating agents in cancer therapy is not well understood. Typical alkylating agents include, but are not limited to: nitrogen mustards, such as, for example, chlorambucil, cyclophosphamide, isofamide, mechlorethamine, melphalan, uracil mustard; aziridine such as, for example, thiotepa; methylsulfonate esters such as, for example, busulfan; nitrous ureas, such as, for example, carmustine, lomustine, est reptozocin; platinum complexes, such as, for example, cisplatin, carboplatin; bioreductive alkylator, such as, for example, mitomycin, and procarbazine, dacarbazine and altretamine. Agents for DNA strand breakage include bleomycin, for example. Shallow topoi II inhibitors of DNA include intercalators, such as, for example, amsacrine, dactomycin, daunorubicin, doxorubicin (adriamycin), idarubicin, and mitoxant scab; non-intercalators, such as, for example, etoposide and teniposide, for example. A minor channel binder of DNA is plica icine, for example. Antimetabolites interfere with the production of nucleic acids by one of two main mechanisms. Certain drugs inhibit the production of deoxyribonucleoside triphosphates, which are the intermediate precursors for DNA synthesis, thus inhibiting DNA duplication. Certain of the compounds are analogues of purines or pyrimidines and are incorporated into the anabolic nucleotide trajectories. These analogs are then replaced in DNA or RNA instead of their normal counterparts. Antimetabolites useful herein include, but are not limited to, folate antagonists, such as, for example, methotrexate and t r imet r exat; pyrimidine antagonists, such as for example, -fluorouracil, fluorodeoxyuridine, CB3717, azacitidine, cytarabine, and floxuridine; purine antagonists include mercaptopurine, 6-thioguanine, fludarabine, pent or s t t ina; and inhibitors of ibonucleotide reductase include idroxyurea. Interactive tubulin agents act by binding to specific sites on tubulin, a protein that polymerizes to form cellular microtubules. Microtubules are essential cellular structural units. When the interactive agents bind to the protein, the cell can not form microtubules. Interactive tubulin agents include vincristine and vinblastine, both alkaloids and paclitaxel (Taxol), for example. Hormonal agents are also useful in the treatment of cancers and tumors. They are used in hormonally susceptible tumors and are usually derived from natural sources. Hormonal agents include, but are not limited to: estrogens, conjugated estrogens and ethinyl estradiol, and diethylstilbesterol, clortrianisen and idenestrol; progestins such as, for example, hydroxyprogeny 3-erona caproate, medroxyprogesterone, and megestrol; and androgens such as, for example, tosterone tes, tosterone testosterone propionate; f luoximes terona, and methyltestosterone. Adrenal corticosteroids are derived from natural adrenal cortisol or hydrocortisone. They are used because of their anti-inflammatory benefits, as well as the ability of some to inhibit mitotic divisions and interrupt DNA synthesis. These compounds include, but are not limited to: prednisone, dexamet asone, methylprednisolone and prednisolone. Hormone agents for luteinizing hormone release or hormonal antagonists for gonadotropin release are used primarily for the treatment of prostate cancer. These include leuprolide acetate and goserelin acetate. They avoid the biosynthesis of spheroids in the testes. For example, antihormonal antigens include antiestrogenic agents such as, for example, tamoxifen, antiandrogenic agents such as, for example, flutamide; and anti-adrenates agents such as, for example, mitotana and aminoglute imide.
Additional agents include the following: hydroxyurea appears to act primarily through the inhibition of the enzyme ribonucleotide reductase, and asparaginase is an enzyme that converts asparagine to non-functional aspartic acid and thus blocks the synthesis of the protein in the tumor. Taxol (paclitaxel) is a preferred chemotherapeutic agent. A list of chemotherapeutic agents currently available according to the class is given as Table 3, and includes the diseases for which the agents are indicated.
Table 3. Neoplastic diseases1 for which the example guimiotherapeutic agents are indicatedClass Type of agent Name Disease2 Agents Mechlorethamine mustard Hodgkin's disease, non-alkylating nitrogen (HN2) lymphomas Hodgkin Cyclophosphamide Acute and chronic lymphocytic leukemia, Ifosfamide Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, neuroblastoma , soft tissue breast, ovarian, lung sarcoma, Wilms tumor, cervix, testicular Mel alán Multiple myeloma, breast, ovary Chlorambucil Chronic lymphocytic leukemia, primary macroglobulinemia, Hodgkin's disease, non-Hodgkin's lymphomas Estramustine Prostate Ethylene imines and Hexamethylmelamine Ovary methylmelamines Thiotepa Bladder, breast, ovary Alkyl sulphonates Busulfan Chronic granulocytic leukemia Nitrosoureas Carmustine Hodgkin's disease, non-Hodgkin's lymphomas, primary brain tumors, multiple myeloma, malignant melanoma Lomustine Hodgkin's disease, non-Hodgkin's lymphomas, brain tumors primary ales, small cell lungs Semustine Primary brain tumors, stomach, colon Streptozocin Malignant pancreatic insulin, malignant carcinoid Triacenes Dacarbazine Malignant melanoma, Hodgkin's disease, Procarbazine soft tissue sarcomas AziridineAntimetabolites Acid analogues Methotrexate Acute lymphocytic leukemia, choriocarcinoma, folic Trimetrexate mycosis fungoides, breast sarcoma, head and neck, pulmonary, osteogenic Fluorouracil analogs Breast, colon, stomach, pancreas, ovary, pyrimidine Floxuridine head and neck, urinary, bladder, premalignant (topical) skin lesions Cytarabine Acute granulocytic and lymphocytic leukemia Azacitidine Purine analogues Mercaptopurine Acute, granulocytic and acute inhibitory lymphocytic and related chronic granulocytic leukemias Thioguanine Acute granulocytic, acute lymphocytic and chronic granulocytic leukemias Pentostatin Hairy cell leukemia, mycosis fungoides, leukemia chronic lymphocytic Fludarabine Chronic lymphocytic leukemia, Hodgkin's and non-Hodgkin's lymphomas, mycosis fungoides Products Vinca Vinblastine's Alkaloids (VLB) Hodgkin's disease, non-natural lymphomas Hodgkin's, breast, testicular Vincristine acute lymphocytic leukemia, neuroblastoma, Wilms tumor, rhabdomyosarcoma, Hodgkin's disease, non-Hodgkin lymphomas, small cell lung Vindesina Acute lymphocytic leukemia * Vinca-resistant, chronic myelocytic leukemia, melanoma, lymphomas, breast Epitodophyllotoxins Etoposide Testicles , small cell lung and other Teniposide, breast, Hodgkin's disease, non-Hodgkin's lymphomas, acute granulocytic leukemia, Kaposi's sarcoma Antibiotics Dactinomycin Choriocarcinoma, Wilms tumor, rhabdomyosarcoma, testicles, Kaposi's sarcoma Daunorubicin Acute lymphocytic and granulocytic leukemiasDoxorubicin Soft tissue sarcomas, Osteogenes Leos, and 4'- others; Hodgkin's disease, lymphocytes that do not Deoxidoxorubicin are Hodgkin's, acute leukemias, breast, genitourinary, thyroid, pulmonary, stomach, neuroblastoma Bleomycin, Testicles, head and neck, skin, esophagus, lung, and genitourinary tract; Hodgkin's disease, non-Hodgkin lymphomas Plicamycin Testicles, malignant hypercalcemia Mitomycin Stomach, cervix, colon, breast, pancreas, bladder, head and neck Enzymes L-Asparaginase Acute lymphocytic leukemia Taxans Docetaxel Mama, ovarian Taxoids Paclitaxel Alpha Interferon modifiers Hairy cell leukemia, aposi sarcoma, melanoma response, carcinoid, renal cell, ovary, biological bladder, non-Hodgkin lymphomas, mycosis fungoides, multiple myeloma, chronic granulocytic leukemia Invasive tumor factor tumor necrosis Lymphocytes for Investigational tumor infiltration Miscellaneous agents Cisplatin complexes Testicular, ovarian, bladder sarcoma, head coordination with Carboplatin and neck , pulmonary, thyroid, cervix, platinum endometrium, neuroblastoma, osteogenic Anthracene Mitoxantrone Acute granulocytic leukemia, breast Replaced urea Hydroxyurea Chronic granulocytic leukemia, polycythemia vera, essential malignant melanoma thrombocytosisDerivative of methyl Procarbazine Hodgkin's disease Hydrazine Suppressor Mitotan Adrenocortical adrenal cortex Aminoglutethimide Breast Hormones and Adrenocortis-Prednisone Acute and chronic lymphocytic leukemias, non-Hodgkin's coesteroid antagonists, Hodgkin's disease, breast Progestins Hydroxy caproate Endometrium, breast Progesterone Acetate of Medroxyprogesterone Megestrol Acetate Estrogens Dietillestil-bestrol Breast, prostate Etinil estradiol Antienstrogen Tamoxifen Mama Androgens Testosterone Mama propionate Fluoxymesterone Antiandrogen Flutamide Prostate Hormone analogue Leuprolide Goserelin Prostate, breast estrogen-recepti "ora- for release of positive gonadotropin Adapted from Calabresi, P ., and BA Chabner, "Chemotherapy of Neoplastic Diseases" Section XII, pp 1202-1263 in: Goodman and Gilman's The Pharmacological Basis of Therapeutics, Eighth ed., 1990 Pergamin Press, Inc .; and Barrows, LR, "Antineoplastic and I mmunoactive Drugs ", Chapter 75, pp 1236-1262, in: Remington: The Science and Practice of Pharmacy, Mack Publishing Co. Easton, PA, 1995; both references are incorporated herein by reference, in particular for treatment protocols. 2Neoplasms are carcinomas unless otherwise indicated.
E. Photoreleasers An "enhancer", in the sense in which it is used herein, is a material that improves or increases the efficacy of the benzimidazole derivatives or a salt or prodrug thereof, or which acts on the immune system. as an immunomodulato. The potentiators can be used in combination with a compound of the present invention. An enhancer can be an antiviral agent. One of these enhancers is triprolidine or its cis-isomer. Triprolidine is described in U.S. Patent 5,114,951 (1992, the patent is incorporated herein by reference). An additional enhancer is procodazole, (also called lH-be z imide z or 1 -2-propanoic acid, or β- (2-benzimidazole) propionic acid or 2- (2-carboxyethyl) benzimide zol or propazole). Procodazole is a nonspecific immunoprotective agent active against viral and bacterial infections and may be used in combination with the compounds set forth herein. Procodazole is effective with a compound of the present invention, only in the treatment of cancers, tumors, leukemia or viral infections, or combined with a chemotherapeutic agent.
Additional potents include, but are not limited to: propionic acid, salts thereof, or esters thereof; antioxidant vitamins such as, for example, vitamins A, C, E, or beta-carotene; abacavir; AL-721 (mixture of lipids); amprenavir; Amphotericin B methyl ester; Ampligen (decoupled RNA); anti-AIDS antibody; interferon-a antibody to i-human; anti-AIDS antibody, ascorbic acid and derivatives thereof; AS-101 (immunostimulants based on heavy metals); a z idotimidine; ß-int erferon; Bupirimine; butylated hydroxytoluene; Ciamexon, Cimetidine; CL-246,738, colony stimulating factors, including G -CSF; Creme Pharmatex (benzalkonium chloride); CS-82 (derived from 5-unsubstituted Zidovudine); Ciclosporin; D-penicillamine (3-mercapto-D-valine); delavirdine; Dextran sulfate; dinitrochlorobenzene; efavirenz; erit ropoyetine;Foscarnet (trisodium phosphonoformate); fusidic acid; ganciclovir; glucan; glycyrrhizin, HPA-23 (ammonium-21-tungs to-9 -antimonate); antiviral human immuno-virus; hyperimmune gamma-globulin, IMREG-1, IMREG-2; indinavir; interferon-a; interferon-gamma; interleukin-1 or int erleucin-2; isoprinosine; Krestin; LC-9018; lamivudine;lentilart LF-1695; methionine-encephalomyelitis; Minophagen C; the tripeptide of muramilo; naltrexone; nelfinavir; Neutropin; nevirapine; Nonoxynol; Ornidyl(eflornit ina); inhibitors without transcript reverse transcript nucleoside; Nucleoside analogues (ddA, ddC, ddl, ddT, ddG, AZT, and the like); pentamidine isethionate; Phenytoin; polyimanoacetate; Peptide T (octapept idica sequence); protease inhibitors; Ribavirin; Rifabutin (ansamycin); ritonavir; AR immunomodulator; rsT4 (recombinant soluble T4); saquinavir; shosaikoto and ginseng; SK-818 (antiviral derived from germanium); sodium diethylcarbonate diet; stavudine; stearic acid derivative; suramin and the like thereof; thymic humoral factor; TP-5; Fraction 5 of Tymosin and Tymosin 1; Timostimulin; TNF (tumor necrosis factor), vitamin B preparations; Tr imet rexato; UA001; al fa-int erferon; c acyclovir, for example. A compound, or a salt or a prodrug thereof, of the present invention can be combined with an enhancer and a chemotherapeutic agent in the methods of the present invention.
F. Dosage Any suitable dosage can be administered in the methods of the present invention. The compound or salt or prodrug thereof selected for a particular application, the carrier and the amount will vary widely depending on the species of warm-blooded animal or human, the type of cancer, or the particular viral infection that will be treated, and it will depend on the effective inhibitory concentrations observed in the analysis studies. The dosage to be administered, of course, will vary depending on known factors, such as, for example, the armacodynamic characteristics of the particular compound, the salt, or combination and its mode and route of administration; the age, health, or weight of the subject; the nature and magnitude of the symptoms; the metabolic characteristics of the drug and the patient, the type of simultaneous treatment; the frequency of treatment; or the desired effect. In general, a dosage as small as approximately 1-2 milligrams (mg) per kilogram (kg) of body weight is suitable, although preferably as little as 10 mg / kg and up to approximately 10,000 mg / kg may be used. . Preferably, a dosage of 15 mg / kg to about 5000 mg / kg is used. More preferably, the dose is between 150 mg / kg to about 1000 mg / kg. The doses useful in the treatment for cancer or viral infections are 250 mg / kg, 500 mg / kg, 800 mg / kg, 1000 mg / kg, 1500 mg / kg, 2500 mg / kg, 3500 mg / kg, 4000 mg / kg, 5000 mg / kg, or 6000 mg / kg. Any variation of dose can be used. In general, a compound, salt thereof, prodrug thereof, or a combination of the present invention can be administered on a daily basis _ one or more times a day, or one to four times a week, either in a single dose or in separate doses during the day. Dosing twice a week for a period of at least several weeks is preferred, and dosing will often continue for prolonged periods of time and possibly for the life of the patient. However, the dosage and dosing regimen will vary, depending on the ability of the patient to sustain the effective plasma levels of the compounds of the present invention, or salt or prodrug thereof, in the blood.
The compound, a salt thereof, prodrug thereof, or a combination can be micronized or pulverized in such a way as to disperse and solubilize more easily by the body. Processes for crushing or spraying drugs are well known in the art. For example, a hammer mill or a similar milling device can be used. The preferred particle size is less than about ??? μ and preferably less than 50μ. Intravenously, the most preferred doses may vary between about 1 and 10 mg / kg / minute during an infusion at constant speed. The dosage for humans in general will be lower than that used in mice and typically is about 1/12 of the dose that is effective in mice. Thus, if 500 mg / kg was effective in mice, a dose of 42 mg / kg could be used in humans. For a 60 kg man, this dose could be 2520 mg. The compounds and salts and prodrugs thereof of the present invention will generally be safe. The LD50 is high, approximately 1500 mg / kg administered orally in mice and there is no special handling requirement. The compounds. they can be administered orally, and because they are not very soluble, they are preferably administered in the form of a tablet or as a suspension. Alternatively, when micronized to a sufficiently small size, they can be administered and preferably do so as an injection. The compounds and salts and prodrugs thereof of the present invention can be administered in a unit dosage form which can be prepared by any method known to one of skill in the art in light of the present disclosure. Unit dosages may include from 1 milligram to 1000 milligrams of the active ingredient. Preferably, the dosage unit will contain between approximately 10 mg and 500 mg of the active ingredient. The active ingredient is generally present in an amount between about 0.5% and 95% by weight based on the total weight of the dosage unit. For intravenous use, preferred dosages may vary between about 1 and 10 mg / kg / minute during an infusion at constant speed.
A dosage unit may comprise a single compound, or mixtures thereof, with other compounds or other cancer inhibitory or viral compounds. The dosage unit may comprise diluents, expanders, carriers, liposomes, or the like. The unit may be in solid form or gel, such as for example, pills, tablets, capsules and the like or in liquid form suitable for oral, rectal, topical, intravenous or parenteral injection or injection in or around the treatment site. The variation and proportion of the benzimidazole derivative, or salt or prodrug thereof, to the chemotherapeutic agent or enhancer will depend on the type of cancer or viral infection being treated and the particular chemotherapeutic agent or enhancer.
G. Formulations Formulations of the present invention include the compound of the present invention, a salt thereof or a prodrug thereof and, optionally, a chemotherapeutic agent and, optionally, an enhancer, generally mixed with a pharmaceutically acceptable carrier. . A "pharmaceutical carrier" is a pharmaceutically acceptable solvent, "suspending agent or vehicle for delivering a compound of the present invention to the animal or human." The carrier can be liquid or solid and is selected in a planned administration manner in mind. A "pharmaceutically acceptable component" is one that is suitable for use with humans and / or animals without causing undue adverse side effects (such as, for example, toxicity, irritation, and allergic responses) in accordance with a reasonable proportion of benefit. / Risk Oral formulations suitable for use in the practice of the present invention include effervescent or non-effervescent capsules, gels, seals, tablets, powders or tablets, powders or granules, as a solution or suspension in aqueous or non-aqueous liquid, or as an oil-in-water liquid emulsion or a water-in-oil emulsion The compounds of the present invention n may also be presented as a bolus, electuary, or paste. In general, the formulations are prepared by uniformly mixing the active ingredient with liquid carriers or finely divided solid carriers or both, and then if necessary shaping the product. A pharmaceutical carrier is selected on the basis of the selected route of administration and normal pharmaceutical practice. Each carrier must be "acceptable" in the sense that it is compatible with the other ingredients of the formulation and does not harm the subject. This carrier can be a solid or liquid and the type in general is selected based on the type of administration to be used. Examples of suitable solid carriers include lactose, sucrose, gelatin, agar and bulk powders. Examples of suitable liquid carriers include pharmaceutically acceptable water, fats and oils, alcohols or other organic solvents, including esters, emulsions, syrups or elixirs, suspensions, solutions and / or suspensions, and reconstituted solutions and / or suspensions from of non-effervescent granules and effervescent preparations reconstituted from effervescent granules. For example, these liquid carriers may contain solvents, preservatives, emulsifying agents, suspending agents, diluents, sweeteners, thickeners, and suitable dissolving agents. Preferred carriers are edible oils, for example, corn oils or canola. Polyethylene glycols, for example PEG, are also preferred carriers. Formulations for oral administration may comprise an inert, pharmaceutically acceptable, non-toxic carrier, such as, for example, lactose, starch, sucrose, glucose, methylcellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol. , cyclodextrin, cyclodextrin derivatives, or the like. The capsules or tablets can be formulated easily and can be easy to swallow or chew. The tablets may contain suitable carriers, binders, lubricants, diluents, disintegrating agents, coloring agents, flavoring agents, flow-inducing agents, or agents for dissolution. A tablet can be made by compression or molding, optionally with one or more additional ingredients. Compressed tablets can be prepared by compressing the active ingredient in a free-flowing form (eg, powder, granules) optionally mixed with a binder (eg, gelatin, hydroxypropylmethylcellulose), lubricant, inert diluent, preservative, disintegrating agent (for example, sodium starch glycolate, crosslinked carboxymethylcellulose) surfactant or dispersant. Suitable binders include starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as, for example, acacia, tragacanth, or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes, or the like. The lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, or the like. Disintegrants include, for example, starch, cellulose, agar, bentonite, xanthan gum, or the like. The molded tablets can be produced by molding in a suitable machine a mixture of the pulverized active ingredient, moistened with an inert liquid diluent. The tablets can optionally be coated or labeled and formulated in order to provide a slow or controlled release of the active ingredient. The tablets may also optionally be provided with an enteric coating to provide release in the parts of the intestine other than the stomach.
Exemplary pharmaceutically acceptable carriers and excipients that can be used to formulate the oral dosage forms of the present invention are described in United States Patent No. 3,903,297 to Robert, issued September 2, 1975, incorporated herein by reference. as reference. The techniques and compositions for preparing the dosage forms useful in the present 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 2nd. Edition (1976). Formulations suitable for topical administration in the mouth, wherein the active ingredient is dissolved or suspended in a suitable carrier include pellets which may comprise the active ingredient in a flavored carrier, typically sucrose and acacia or tragacanth; gelatin, glycerin, or sucrose and acacia; and mouth rinses comprising the active ingredient in a suitable liquid carrier. Topical applications for administration according to the method of the present invention include ointments, cream, suspensions, lotions, powder, solutions, pastes, gels, spray, aerosol or oil. Alternatively, a formulation may comprise a transdermal patch or bandage, such as, for example, a bandage impregnated with an active ingredient and optionally one or more carriers or diluents. To be administered in the form of a transdermal delivery system, dosing administration will, of course, be continuous rather than intermittent throughout the dosing regimen. Topical formulations may conveniently include a compound that reinforces the absorption or penetration of the active ingredient through the skin or other affected areas. Examples of these dermal penetration sensors include dimethylsulfoxide and related analogues. The oil phase of an emulsion used to treat subjects in the present invention may be constituted of ingredients known to one of skill in the art in the light of the present disclosure. An emulsion may comprise one or more emulsifiers. For example, an oil phase may comprise at least one emulsifier with a fat or an oil, either with a fat or with an oil, or a hydrophilic emulsifier may be included together with a lipophilic emulsifier which acts as a stabilizer. Together, the emulsifiers, with or without stabilizers, constitute an emulsifying wax, and the wax together with the oil and / or grease constitute the emulsifying ointment base which forms the oily dispersed phase of the cream formulations. Emulsifiers and emulsion stabilizers suitable for use in the formulation include T een 60, Span 80, ketoestilic alcohol, myristyl alcohol, glyceryl monostearate and sodium lauryl sulfate, paraffin, mono or dibasic alkylesters, straight or branched chain, oil mineral. The choice of oils or greases suitable for the formulation is based on achieving the desired cosmetic properties, the required properties and compatibility with the active ingredient. The compounds of the present invention can also be administered vaginally, for example, as pessaries, tampons, creams, gels, pastes, foams or spray formulations containing suitable carriers in addition to the active ingredient. These carriers are known in the art in light of the present disclosure.
Formulations for rectal administration may be presented as a suppository with a suitable base comprising, for example, cacao butter or a salicylate. Formulations suitable for nasal administration may be administered in a liquid form, for example, nasal spray, nasal drops, or by administration of the aerosol by a nebulizer, which includes aqueous or oily solutions of the active ingredient. Formulations for nasal administration, wherein the carrier is a solid, includes a coarse powder having a particle size, for example, of less than about 100 microns, preferably less than about 50 microns that is administered in the manner in which it is administered. snuff is taken; that is, by rapid inhalation through the nasal passage of a powder container held close to the nose. Formulations suitable for parenteral administration include aqueous and non-aqueous formulations isotonic with the blood of the intended recipient; and sterile aqueous and non-aqueous suspensions which may include suspension systems designed to direct the compound to the blood components or one or more organs. The formulations can be presented in sealed unit dose or multidose containers, for example, ampoules or vials. Solutions and suspensions may be prepared in extemporaneous injections, from sterile powders, granules and sterile tablets of the kind described above. The forms• Parenteral and intravenous can also include minerals and other materials to make them compatible with the type of injection or delivery system selected. In general, suitable carriers for parenteral solutions are water, a suitable oil, saline solution, aqueous dextrose (glucose), or related sugar solutions and glycols such as, for example, propylene glycol or polyethylene glycols. Solutions for parenteral administration preferably contain a water soluble salt of the active ingredient, suitable stabilizing agents and, if necessary, buffering substances. Suitable stabilizing agents are antioxidant agents, such as, for example, sodium bisulfite, sodium sulfite, or ascorbic acid, alone or in combination. Citric acid salts thereof, or sodium EDTA, are also used. In addition, parenteral solutions may contain preservatives, such as, for example, benzalkonium chloride, methyl- or pr-opyl-paraben, or chlorobutanol. Suitable pharmaceutical carriers are described in Remington, cited above. The present invention additionally contemplates administering the compounds of the present invention described in the invention for use in the form of veterinary formulations which can be prepared, for example, by methods that are conventional in the art in light of the present disclosure. Pharmaceutical dosage formulations useful for the administration of the compounds of the present invention are illustrated as follows: Capsules: many unit capsules are prepared by filling two-piece normal hard gelatin capsules, each with 100 milligrams of powdered active ingredient, 150 milligrams of lactose, 50 milligrams of cellulose, and 6 milligrams of magnesium stearate. Soft gelatine capsules: A mixture of the active ingredient is prepared in a digestible oil, such as, for example, soybean oil, cottonseed oil or olive oil and is injected by means of a positive displacement pump into the interior of Gelatin to form soft gelatin capsules containing 100 milligrams of the active ingredient. The capsules are washed and dried. Tablets: Many tablets are prepared by conventional procedures so that the dosage unit is 100 milligrams of active ingredient, 0.2 milligrams of colloidal silicon dioxide, 5 milligrams of magnesium stearate, 275 milligrams of microcrystalline cellulose, 11 milligrams of starch and 98.8 milligrams. milligrams of lactose. Suitable coatings can be applied to increase palatability or delay in absorption. Injectable: A parenteral composition suitable for administration by injection is prepared by stirring 1.5% by weight of active ingredient in 10% by volume of propylene glycol and water. The solution is made isotonic with sodium chloride and sterilized. Suspension: An aqueous suspension is prepared for oral administration in such a way that every 5 ml contains 100 mg of finely divided active ingredient, 200 mg of sodium carboxymethylcellulose, 5 mg of sodium benzoate, 1.0 g of sorbitol solution, USP, and 0.025 ml of vanillin. The compounds of the present invention can be administered in the form of liposomal delivery systems, such as, for example, small unilamellar vesicles, large unillar amelar vesicles, and. multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as, for example, cholesterol, stearylamine, or phosphate idylcholine. The compounds of the present invention can be coupled with soluble polymers such as for example, white drug carriers. These polymers can include polyvinylpyrrolidone, pyran copolymer, polyhydroxylpropylmeta-crilamide-phenol, polyhydroxyethylaspartamidephenol, or polyethyleneoxide-polylysine substituted with palmitoyl residues. In addition, the compounds of the present invention can be coupled to a class of biodegradable polymers useful for achieving controlled release of a drug, for example, polylactic acid, polyglycolic acid, polylactic and polyglycolic acid copolymers, polysilon caprolactone, polyhydroxybutyl acid l, poliortoes tere s, polyacetals, polydihydropyrans, polycyanoacrylates, and cross-linked or amphipathic block copolymers of hydrogels.
H. Treatment method The treatment method can be any suitable method that is effective in the treatment for the particular cancer or viral infection being treated. The treatment includes administering a therapeutically effective amount of the compounds of the present invention in a manner described above to a subject in need of treatment. The compounds of the present invention can be administered by any means that produces contact of the active agent with the site of action of the agent in the body, for example, suitable means include, but are not limited to: oral, rectal, nasal, topical (including t ransdermal, aerosol, buccal or sublingual), vaginal, parenteral (including subcutaneous, intramuscular, intravenous or intradermal.), intravesical, or injection into or around the cancer or site of viral infection. conventional medium available for use in conjunction with pharmaceuticals, either as individual therapeutic agents or in a combination of therapeutics Preferably, the compounds of the present invention are administered as a pharmaceutical formulation comprising at least one compound of the present invention, as was defined above, together with one or more pharmaceutically acceptable carriers. they can co-administer in the form of a tablet or capsule, as an agglomerated powder, or in a liquid form, or as a liposome. The preferred route will vary with the condition and age of the recipient, virus or cancer being treated, the nature of the disorder, or the severity of the disorder. It is believed that oral administration, or parenteral treatment is the preferred method for administering the compounds to subjects in need thereof. In each of the methods described above, the administration can be in vivo, or it can be ex vivo. In vivo treatment is useful for treating diseases in a mammal, preferably, the mammal is a human being; and the ex vivo treatment is useful for purging bodily fluids, such as, for example, blood, plasma, bone marrow, and the like, to return them to the body. The nation's blood supply is commonly tested for antibodies to HIV. However, the test is still imperfect and samples that provide negative tests may still contain the HIV virus. Blood for treatment and blood products with the compounds of the present invention can add an extra margin of safety to kill any retroviruses that may not have been detected. The body tissue can be internal or external to an animal body, or, for example, it can be the superficial skin of the animal.
I. Combination Therapy The compounds of the present invention can be further combined with chemotherapeutic agents and / or enhancers to provide combination therapy. The combination therapy is intended to include any chemically compatible combination of a compound of the present invention with other compounds of the present invention or other compounds outside the present invention, provided that the combination does not eliminate the activity of the compound of the present invention. invention. For example, one or more compounds can be combined with an enhancer or chemotherapeutic agent. In the case of a retroviral infection; The present invention contemplates a combination therapy with nucleoside analogues such as AZT, inhibitors of nucleoside reverse transcriptase, TC-3, or protease inhibitors. In the case of hepatitis, it is contemplated as a combination therapy acyclovir, famciclovir or valaciclovir, Ribavirin, interferon, or combinations of Ribavirin and interferon or beta globulin. For herpes, a recombinant alpha interferon can be used as a combination therapy. The active agent can be co-administered, for example, in the form of a tablet or capsule, liposome, as an agglomerated powder, or in a liquid form. The amount of the chemotherapeutic agent or enhancer used may be as low as that of the benzimidazole derivative. It will be present in a dosage unit in an amount that provides an operational combination with the benzimidazole derivative. The dosage of the chemotherapeutic agent or enhancer may vary between about 0.5 mg / kg of body weight and 400 mg / kg of body weight.
The combination therapy can be sequential, that is, the treatment with a first agent and then with the second agent, c the treatment can be with both agent at the same time. Sequential therapy may be within a reasonable time after completing the first therapy - before starting the second therapy. The treatment with both agents at the same time can be in the same daily dose or in separate doses. For example, treatment with one agent on day 1 and the other on day 2. The exact regimen will deplete the disorder being treated, the severity of the disorder, and the response to treatment. In addition to the use of chemotherapeutic agents and enhancers, a benzimidazole derivative or a salt or a prodrug thereof can be combined with a fungicide or a herbicide. Preferred herbicides and fungicides include carbendazim, fluoconazole, benomyl, glyphosate, and propicodazole.
J. Pharmaceutical equipment The present invention also includes pharmaceutical kits useful, for example, for the treatment of cancer or a viral infection. The kits comprise one or more containers containing a pharmaceutical composition comprising a therapeutically effective amount of a compound of the present invention. These kits may additionally include, if desired, for example, one or more of various components of the conventional pharmaceutical equipment, such as, for example, containers with one or more pharmaceutically acceptable carriers, additional containers, etc., as will be readily apparent by those experts in the art. Instructions such as, for example, printed instructions, either as inserts or labels, quantities indicating instructions for components to be administered, guidelines for administration, and / or guidelines may also be included on the equipment. to mix the s components.
K. Studies The following studies were conducted to test the efficacy of the benzimidazole derivatives of the present invention against certain cancers and viral infections.
Molecular colon and melanoma t cell test: The following cell culture tests were performed to test the toxicity of the benzimidazole compound derivatives of the present invention in colon and melanoma tumor cells. The viability of the cells was tested by observing the reduction of TT (3- [4,5-dimethylthiazol-2-yl] -2,5-diphenyltetrazolium bromide). MTT analysis is a well-known measurement of cell viability. Colon tumor cells (HT29). and the melanoma cells (B16 murine melanoma) were seeded (1000-2000 cells) in each well of a 96-well fflicrot itulator plate and allowed to grow. Twenty-four hours later, increasing concentrations of each benzimidazole derivative were added to the plates and the cells were allowed to incubate for 3-6 days in the presence of the drugs. The MTT reagent was then added to the wells and allowed to incubate for 4 hours at 37 ° C after the formazan metabolite was solubilized with acid isopropanol and the absorbance was read. The experimental controls included empty cavities to which no cells were added (zero point of formazan absorbance) and control cavities to which no drug was added (highest level of formazan absorbance). Each drug concentration was tested in duplicate and the resulting average absorbance plotted against the drug concentration using the EZ-ED50 computer program on a personal computer connected to the microtiter plate reader. The program adjusted the resulting curves to the four parameter equation:where Amax and Amin are, respectively, the absorbances in the absence and presence of the highest drug concentration, X is the concentration of drug and Y is the absorbance at that concentration, n is the slope of the curve, and IC50 is the concentration of drug that provides 50% inhibition of growth. The IC50 value is then derived from this equation by EZ-ED50. The IC50 values for the inhibition of growth of cancer cell lines both B16 and HT29 are reported in the following Table 4 (Y = H unless otherwise indicated).
TABLE 4 MTT ANALYSIS - GROWTH INHIBITOR ACTIVITY Cl, in position 5 (6) Microtubule inhibition analysis: The benzimidazole derivatives of this invention were evaluated for their ability to inhibit the formation of microtubules, following the procedure of Luduena, RF and MC Roach (Pharmacol, Ther 49: 133-152 (1991)). Tubulin was purified from bovine brain by a cycle of assembly and disassembly, followed by phosphorus focellulose chromatography and gel filtration (Fellous, A. et al., Eur. J. Biochem. 78 (1): 167-174 ( 1977)). The analysis was performed by preparing a sample of tubulin protein under conditions that stimulate microtubule polymerization (Prasad, V. et al., J. Protein Chem. 11 (5): 509-515 (1992)), then adding the derivatives of benzimide zo 1, to 2μ ?, and following the chronological course of polymerization. The microtubular polymerization was followed when performing the reaction in a cuvette and monitoring the UV absorbance at 350 nrti, which allows direct visualization of the chronological course of the polymerization. After 30 minutes, the polymerization was completed and the polymerization inhibition percentage was calculated as the final absorbance of the reaction taken as a percentage of the final absorbance of a control run parallel polymerization reaction without added drug. These data are reported in the following Table 5.
TABLE 5 INHIBITION OF TUBULINE POLYMERIZATION* Y = Cl, in position 5 (6) DNA binding analysis: DNA binding analysis was performed in a manner very similar to that of the MTT cell growth inhibition analysis above. The analysis was based on the ability of the test compounds to displace the methyl green (Sigma) from the DNA and the resulting hydration of the free methyl green molecule to a colorless derivative (Burres, NS et al., J. Nat. Prod. 55 (11): 1582-1587 (1992), Kim and Nordén, FEBS Lett 315 (1): 61-64 (1993)). Regarding the MTT analysis, each drug concentration was analyzed in duplicate and the data were averaged before the analysis. The experimental controls in this analysis included cavities containing free methyl green solution without DNA, for the totally colorless previous absorbance, and methyl green-DNA complex (Sigma) without drug, for the highest absorbance of methyl green. · The methyl-DNA green in the buffer was added to each well to obtain A655 of about 0.7. Serial dilutions of drug were added to the buffer and the plates were mixed, then allowed to stand at room temperature for 24 hours to allow displacement and hydration of free methyl green. The absorbance at 655 mm was measured as a function of aggregate drug concentration in an automated microplate reader, and the data was reduced by the EZ-ED50 computer program to derive IC50 values (concentration at which 50% of the green of methyl was displaced from the DNA). None of the benzimidazole derivatives listed in Table 4 or 5 showed any DNA interaction by this analysis.
EXAMPLES Example 1: Summary of Comp. No. 1-4 0.200 grams (1.5 mmol) of 2-aminobenzimide zol were dissolved in 4 mL of dehydrated pyridine and one equivalent (1.5 mmol, 200 mg) of benzyl isocyanate was added under argon. The mixture was stirred for 16 hours under argon at 23 ° C and then another equivalent of benzyl isocyanate was added. After an additional four hours of stirring under argon at 23 ° C, the mixture was emptied into 20 mL of water to precipitate the product. The product was collected by filtration, washed with water and dried under reduced pressure for 16 hours to provide 0.234 g (0.879 mmol, 58.6% yield) of white powder.
E em lo 2: Synthesis of Comp. No. 1-5 0.200 grams (1.5 mmol) of 2-aminobenzimidazole were dissolved in 4 mL of dehydrated pyridine and one equivalent (1.5 mmol, 256 mg) of benzyl chloroformate under argon was added. The mixture was stirred for 16 hours under argon at 23 ° C and then another equivalent of benzyl chloroformate was added. After an additional four hours of stirring under argon at 23 ° C, the mixture was emptied into 20 mL of water to precipitate the product. The product was collected by filtration, washed with water and dried under reduced pressure for 16 hours to provide 0.168 g (0.629 mol, 41.9% yield) of white powder.
Step 3: Synthesis of Comp. No. 2-11 0.112 grams (0.421 mmol) of Comp. No. 1-4, of Example 1, were dissolved in 1 mL of dehydrated pyridine and heated at 80 ° C for 5 hours with stirring. Six mL of water was added to the mixture with stirring and the solid precipitate was collected by filtration, washed with water, and dried under reduced pressure for 16 hours. 0.074 g (0.277 mmol, 65.8% yield) of a whitish powdery product were obtained.
Example 4: Synthesis of Comp. No. 2-9 0.040 grams (0.150 mmol) of Comp. No. 1-5, of Example 2 were dissolved in 4 mL of toluene and heated to 110 ° C with stirring for 24 hours.
The mixture was cooled in an ice water bath and the precipitated solid was collected by filtration to provide 0.024 g (0.0.90 mmol, 60% yield) of a product as a white powder.
Example 5: Synthesis of Comp. No. 2-7 0.025 grams (0.131 mmol) 2-aminobenzimidazole methyl carbamate were mixed with 2 mL of toluene and 0.696 g (6.55 mmol) of diethylene glycol. Three mg of aluminum isopropoxide was added and the mixture was heated at 110 ° C for 15 hours. The toluene was evaporated under an argon stream with continuous heating, then it was removed from the heat and the residue was treated with 4 mL of boiling water. The resulting mixture was cooled to 23 ° C, then stored for 18 hours at 4 ° C. The precipitate that formed was collected by filtration and dried under reduced pressure to provide 0.029 g (0.109 mmol, 83.2% yield) of a product as whitish flakes.
Example 6: Synthesis of Comp. No. 2-5 0.025 grams (0.131 mmol) of methyl 2-aminobenzimidazole methyl carbamate were mixed with 2 mL of toluene and 0.817 g (6.55 mmol) of 2- (2-chloroethoxy) ethanol. Three mg of aluminum isopropoxide was added and the mixture was heated at 110 ° C for 15 hours. The toluene was evaporated under an argon stream with continuous heating, then it was removed from the heat and the residue was treated with 4 ml of boiling water. The resulting mixture was cooled to 23 ° C, then stored for 18 hours at 4 ° C. The precipitate that formed was collected by filtration and dried under reduced pressure to provide 0.026 g (0.09 mmol, 68.7% yield) of product as a whitish powder.
Example 7: Synthesis of Comp. No. 2-4 0.025 g of (0.131 mmol) of methyl 2-aminobenzimidazole carbamate were mixed with 2 ml of toluene and 0.689 g (6.55 mmol) of 2- (2-aminoethoxy) ethanol. Three mg of aluminum isopropoxide was added and the mixture was heated at 110 ° C for 15 hours. The toluene was evaporated under an argon stream with continuous heating, then it was removed from the heat and the residue was treated with 4 ml of boiling water. The resulting mixture was cooled to 23 ° C, then stored for 18 hours at 4 ° C. The precipitate that formed was collected by filtration and dried under reduced pressure to provide 0.009 g (0.034 mmol, 25.9% yield) of the product as colorless flake-like crystals.