MXPA99006793A - Terbenzimidazoles useful for medical therapy (topoisomerase inhibitors) - Google Patents

Abstract

The invention provides a topoisomerase poision of formula (I) wherein Ar is aryl or a nitrogen-, sulfur- or oxygen-containing heteroaromatic group;X is H, CN, CHO, OH, acetyl, CF3, O(C1-C4)alkyl, NO2, NH2, halogen or halo-(C1-C4)alkyl;each Y is individually H, (C1-C4)alkyl or aralkyl;Y'is phenyl, or methoxyphenyl;n is 0 or 1;and each Z is individually H, (C1-C4)alkyl, halogen or halo(C1-C4)alkyl;or a pharmaceutically acceptable salt thereof;for use in medical therapy (e.g. the treatment of fungal infection or cancer). The invention also provides novel compounds of formula (I);pharmaceutical compositions comprising compounds of formula (I);and therapeutic methods, comprising treating fungal infection or treating cancer by administering at least one compound of formula (I).

Description

USEFUL TERBENCIMIDAZOLES FOR MEDICAL THERAPY (TOPOISOMERASE INHIBITORS) Background of the Invention This invention was made with the support of United States National Institutes of Health Grant CA 39962. The Government of E.U.A. has certain rights in the invention. DNA topoisomerases are nuclear enzymes that control and modify DNA topological states by catalyzing the breakdown and coordinated binding of DNA strands. See, for example, D'Arpa and others, Biochim, Biophys. Acta. 989 (1989). Topoisomerase II enzymes alter the topological state of DNA by means of a double strand break in DNA. Interfering with the rupture / reunion reaction of DNA topoisomerases, a number of agents have been shown to convert these enzymes into network enzymes that break DNA, resulting in efficient cell death. See, L.F. Liu, in Topoisomerases: topoisomerase targeting drugs, Adv. in Pharmacol., 29B (1994); L.K. Wang et al., Chem. Res. Toxicol. 6, 813 (1993). Therefore, mammalian topoisomerase II represents an effective pharmacological target for the development of cancer chemotherapy (A. Y. Chem et al., Annu., Rev. Pharmacol. Toxicol., 34, 191 (1994)). Among the clinical agents in use that are recognized as topoisomerase II inhibitors are etoposide (VP-16), teniposide (VM-26), mitoxantrone, m-AMSA, adriamycin (doxorubicin), ellipticine, and daunomycin.

Compared with topoisomerase II inhibitors, there are relatively few known topoisomerase I inhibitors. Camptothecin represents the mammalian topoisomerase I inhibitor most extensively studied. See R. Gallo and others, sL Nati. Cancer Inst .. 46,789 (1971) and B. C. Giovanella et al., Cancer Res., 51, 3052 (1991). Camptothecin interference with the topoisomerase I cleavage / gap reaction results in the accumulation of a covalent intermediate, in which topoisomerase I is reversibly trapped in a separate state, called the separable complex (Y. -H. Hsiang et al, J. Biol. Chem., 260, 14873 (1985), SE Porter et al., Nucí Acids Res .. 17, 8521 (1989); C. Jaxel et al., J. Biol. Chem. 266, 20418 (1991)). The broad spectrum of potent antineoplastic activity observed for camptothecin has further efforts to identify other agents that can effectively poison mammalian topoisomerase I. It has recently been shown that 2 '- (4-ethoxyphenyl) -5- (4-methyl-1-piperazinyl) -2,5'-bi-1 H-benzimidazole from Hoechst 33342 (1), is a topoisomerase I inhibitor. .

This agent, which binds to the minor DNA groove, traps the reversible separable complex derived from DNA and topoisomerase as well as producing a limited number of highly specific single-stranded DNA breaks. For example, see A. Y. Chem et al., Cancer Res. 53, 1332 (1993) and A. Chem et al., PNAS. 90. 8131 (1993). A limitation of Hoechst 33342 as an anticancer agent is the previously reported observation that it is not effective against tumor cell lines that overexpress MDR1. While KB 3-1 cells are known to be very sensitive to Hoechst 33342, with an IC5o of about 9 nM, this compound is about 130 times less cytotoxic than KB V-1 cells, which is known to overexpress MDR1. Recently, several analogs of this bisbenzimidazole have been synthesized to further investigate the structure activity ratios associated with their potency as inhibitors of mammalian topoisomerase I and related cytotoxicity. For example, Q. Sun and others, Biorg. and Med. Chem. Lett. 4, 2871 (1994) described the preparation of bis-benzimidazoles of the formula (2): where n is 0, 1, 2 or 3. However, these compounds were found to be approximately on the order of magnitude less cytotoxic than Hoechst 33342. More recently, Q. Sun and others, in Abstract 2688, Scientific Proceings-86th Annual Meeting of the AACR (Toronto, CA, March 18-22, 1995) described that a trisbenzimidazole derivative, 5- (2-pyridyl) -2- [2'-benzimidazole-5"-M-benzimidazol-5'-I] benzimidazole having similar potency that a human topoisomerase I inhibitor such as Hoechst 33342. Fungal infections have increasingly become more important in the last two decades, causing high mortality among immunocompromised patients, such as transplant recipients and patients with cancer and SI DA. of patients and some existing problems in current antifungal chemotherapy, have created a demand for more effective and safe antifungal agents for the treatment of this increasingly important class of opportunistic infections Based on studies of Saccharomyces cerevisiae and Candida albicans, topoisomerase I Nuclear fungal shows promise as a molecular target for antifungal agents (see JM Fostel et al., Antimicrob Agents Chemother., 39 586 (1995); J. M. Fostel and others, Antimicrob. Agents Chemother .. 39. 2131 (1992)). Studies in S. cerevisiae have established that topoisomerase I is a fungicidal target for camptothecin (J. Nitiss et al., P NAS USA. 85, 7501 (1988)). Studies in C. albicans have shown differences in the sensitivity of topoisomerase I of humans and Candida to aminocatechol A-3253 (J. M. Fostel (1995) cited above). Aspergillus fumigatus and A. niger are two important systemic human pathogens that threaten life. There is an urgent need for more effective antifungal agents for the treatment of patients with these opportunistic infections. SUMMARY OF THE INVENTION The invention provides a therapeutic method for the treatment of a fungal infection comprising administering to a mammal suffering from a fungal infection, particularly, a systemic fungal infection, an effective antifungal amount of a compound of general formula (I). ): (I) wherein Ar is a heteroaromatic group containing aryl or a nitrogen, sulfur, or oxygen; X is H, CN, CHO, OH, acetyl, CF3, O- C alquilo-C alkyl, NO 2, N H 2, halogen or C halo-C 4 alkyl halo; each Y is individually H, C? -C alkyl or aralkyl; Y 'is H, C? -C alkyl, phenyl or methoxyphenyl; each Z individually is H, C? -C4 alkyl, halogen or C? -C4 haloalkyl; and n is 0 or 1; or a pharmaceutically acceptable salt thereof. Preferably, Ar is a C6-C aryl? 2, such as phenyl or a 5- to 13-membered heteroaryl group, more preferably a 5-6-membered heteroaryl group, comprising 1 -3 N, S or O without peroxide, wherein each N is not substituted or substituted with H, C? -C alkyl or benzyl. Ar may occupy position 4, 5, 6 or 7 of the benzo ring, as shown, preferably position 5 and x may occupy any position available on Ar. Positions 4, 7 and 5, 6 are equivalent when Y is H. According to one embodiment, Ar is phenyl and X is C l or Br, preferably occupying the para position. As plotted, Z can occupy any position in the benzo portion. Preferably Z is H, halogen, CH 3 or CF 3. According to another embodiment, n is 0 and x is halogen, for example, F, Br, Cl or I, preferably Cl or Br, and preferably occupies position 5 of the benzo moiety. And preferably it is H or CH3. Y 'is preferably H, CH 3, ethyl or 4-methoxyphenyl. While a number of known inhibitors of human topoisomerase I were found effective against a fungal topoisomerase I, including nitidine and coralline, the compounds of formula (I) are inhibitors of fungal topoisomerase I, as demonstrated by their ability to promote separation of DNA in the presence of Aspergillus topoisomerase I. As described below, it was unexpected to find that the Aspergillus enzyme is completely resistant to some of the more potent human topoisomerase I poisons such as nitidine and coralline, and to the less potent human mono-benzimidazole topoisomerase I poisons. Studies using human topoisomerase I or yeast expressing yeast also suggest similar resistance to yeast topoisomerase I to these compounds. It seems that the fungal enzymes are substantially different in their sensitivity to drugs than their human counterparts. In addition, the compounds of the formula (I) are also cytotoxic for mammalian tumor cells, including camptothecin-resistant and camptothecin-resistant tumor cells and cell lines that exhibit multidrug resistance due to the expression of P-glycoprotein. . Accordingly, the invention provides a therapeutic method for the treatment of cancer which comprises administering to a mammal (i.e., a human), an effective anticancer amount of a compound of the formula (I), or a pharmaceutically acceptable salt thereof. The invention also provides novel compounds of the formula (I). For example, the invention provides a compound of the formula (I): (I) wherein, Ar is an aryl of C6-C? 2, or heteroaryl of 5 to 13 members, comprising 1-3 N, S or O without peroxide, wherein each N is not substituted or substituted with H, alkyl of C? -C4 or benzyl; X is H, CN, CHO, OH, acetyl, CF3, O-C1-C4 alkyl, NO2, NH2, halogen or haloalkyl of C? -C4; each Y is H, C? -C4 alkyl or aralkyl; Y 'is phenyl or methoxyphenyl; each Z is individually H, C? -C4 alkyl, halogen or C? -C4 haloalkyl; and n is 0 or 1; or a pharmaceutically acceptable salt thereof. A preferred compound is a compound of the formula (I) wherein Y 'is methoxyphenyl. Another preferred compound is a compound of the formula (I) wherein n is 1. Another preferred compound is a compound of the formula (I) wherein X is CN, CHO, OH, acetyl, CF3, O-C alkyl? -C, NO2, NH2, halogen or haloalkyl of C? -C4 and n is 0. Yet another preferred compound is a compound of the formula (I) wherein at least one of Z is halogen or haloalkyl of C? -C and n is 0 The invention also provides pharmaceutical compositions adapted for systemic and topical administration, comprising one or more of the compounds of the formula (I) or a pharmaceutically acceptable salt thereof, in combination with a pharmaceutically acceptable carrier. The invention also provides a compound of the formula (I), or a pharmaceutically acceptable salt thereof for use in medical therapy (ie to treat fungal infections or cancer), as well as the use of a compound of the formula (I), or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament to treat fungal infections or to treat cancer. Brief Description of the Drawings Figure 1 is a schematic description of the synthesis of compounds 10-16. Figure 2 is a schematic description for the preparation of intermediates 4-8 used to prepare compounds of the invention. Figure 3 is a schematic description of the preparation of intermediate 9. Figure 4 is a schematic description of the synthesis of compounds JSKIV-68, 37 and 47. Figure 5 is a schematic description of the preparation of the intermediate JSKIV-44.

Figure 6 is a schematic description of the preparation of the modified analogues in the central benzimidazole moiety. Figure 7 is a schematic description of the preparation of modified analogs in the terminal benzimidazole moiety, wherein Z and Y 'are as defined above. Figure 8 summarizes the activity of several agents against topoisomerase I of humans and Aspergillus. The poisoning activity of several drugs against humans (column H) and Aspergillus (column A) are qualitatively indicated by a + (active) or - (inactive). DM / l / 33 is only very weakly active against Aspergillus topoisomerase I and is indicated by *. Detailed Description of the Invention The (Ar) groups useful in the present compounds comprise C 6 -C 8 aryl, preferably C 6 -C 4 aryl, v. gr. , systems containing aromatic rings, said systems comprise a total of 6 to 12 carbon atoms. Therefore, as used herein, the term "aryl" includes aryl substituted with mono or bis-C 1 -C 4 alkyl, such as tolyl and xylyl; an alkyl of Ci- C4, such as benzyl or phenethyl; and alcaralkyl. Preferably aryl is phenyl, benzyl or naphthyl. Heteroaromatic rings include aromatic rings containing up to 3 ring heteroatoms such as N, S or O without peroxide and up to 12 ring atoms. Representative aromatic rings include thiophene, benzothiophene, naphthothiophene, triantreno, furan, benzofuran, isobenzofuran, pyran, chromene, xanthene, phenoxathiin, pyrrole, imidazole, pyrazole, pyridine, pyrazine, triazole, tetrazole, pyrazine, triazine, pyrimidine, pyridazine, indolizine , isoindole, indole, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, phenanthroline, phenazine, isothiazole, phenothiazine, oxazole, isoxazole, furazane, phenoxazine and similar. Preferred heteroaromatic rings have a 5-6 membered heteroaromatic ring which may or may not be fused to an aromatic ring such as a benzo ring, eg, the preferred 2, 3 or 4-pyridyl substitutents. The term "alkyl" includes straight or branched chain alkyl, as well as cycloalkyl and (cycloalkyl) alkyl, e.g., methyl, ethyl, i-propyl, cyclopropyl or cyclopropylmethyl. Methoxyphenyl includes 2-, 3- or 4-methoxyphenyl. Pharmaceutically acceptable salts include the acid addition with basic NH with organic or inorganic acids, eg, hydrochloride, carbonate, sulfate, bicarbonate, acetate, phosphate, tartarate, citrate, malate, maleate and the like propionate . The preparation of representative substituted trisbenzimidazoles is described in Figure 1. Except for phenylenediamine which was commercially available, phenylenediamines appropriately substituted by catalytic hydrogenation of the respective o-nitroaniline derivatives were synthesized. These phenylenediamines were coupled with 5-formyl-2- (benzimidazo-5'-yl) benzimidazole, 9, heating them together with nitrobenzene at 150 ° C to provide the different trisbenzimidazoles, 10-16, in yields ranging from 43-96% , using the general MP methodologies Singh et al., Chem. Res. Toxicol., 5, 597 (1992) and Y. Bathini et al., Svnth Comm. 20, 955 (1990). The required nitroanilines, as described in Figure 1, except for 3 that was commercially available, were synthesized from 4-bromo-nitroaniline, V7. Compound V7 was prepared from o-trinoaniline in good yield, 94%, using 2,4,4,6-tetrabromo-2,5-cyclohexadienone as the bromination reagent. G.J. Fox and others, Org. Syn., 55, 20 (1973). While allyltributyltin and phenyltributyltin are commercially available, the pyridyltributyltin derivatives were prepared from tributyltin chloride and 2-, 3-, and 4-bromopyrin, respectively. See D. Peters et al., Heterocvclic Chem. 27 2165 (1990). These tributyltin derivatives were coupled with 4-bromo-2-nitroaniline using PdCI2 (PPh3) 2 as the catalyst in DMF as described in Figure 2 to provide compounds 4, 5, 6, 7 and 8, respectively, according to with the methodology of M. Iwao and others, Heterocvcles. 36, 1483 (1993). This methodology can generally be applied to prepare 2-nitroanilines substituted with 3-, 4-, 5-or 6-aryl and heteroaryl of the corresponding bromonitroanilines. The preparation of 5-formyl-2- (benzimidazo-5'-yl) benzimidazole, 9, was achieved as described in Figure 3. The reduction of 5-benzimidazolecarboxylic acid to 5-hydroxymethylbenzimidazole was achieved using LiAIH4. Oxidation of the resulting crude benzyl alcohol with tetrapropylammonium perruthenate (TPAP) and N-methylmorpholine N-oxide provided in two steps 5-formylbenzimidazole desired with an overall yield of 32%. See, A. Cherif et al., J. Med. Chem. , 35, 3208 (1992). The coupling of 5-formylbenzimidazole with 4-cyano-1,2-phenylenediamine provided 5-cyano-2- (benzimidazol-5'-yl) benzimidazole, 19 which, when treated with Ni-AI catalyst in the presence of aqueous formic acid, gave 5-formyl-2- (benzimidazol-5'-yl) benzimidazole, 9, in 65% yield. (J. R. Pipier et al., J ^ Med. Chem. 31, 2164 (1988)). The compounds of the present invention can be formulated as pharmaceutical compositions and administered in a mammalian host, such as an immunosuppressed human patient suffering from a systemic or local fungal infection, in a variety of forms adapted to the chosen route of administration, is say, by oral or parenteral, intravenous, intramuscular, topical or subcutaneous routes. Therefore, the compounds herein can be administered systemically, v. g r. , orally, in combination with a pharmaceutically acceptable carrier such as an inert diluent or an edible assimilable carrier. They can be enclosed in hard or soft shell gelatin capsules, they can be compressed into tablets or they can be incorporated directly with the food of the patient's diet. For oral therapeutic administration, the active compound can be combined with one or more excipients and used in the form of non-digestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers and the like. Said compositions and preparations should contain at least 0.1% active compound. The percentage of the compositions and preparations, of course, can vary and conveniently can be between about 2 to about 60% by weight of a given unit dosage form. The amount of active compound in said therapeutically useful compositions is such that an effective dose level will be obtained. Tablets, troches, pills, capsules and the like may also contain the following: binders such as gum tragacanth, acacia, corn starch or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as corn starch, potato starch, alginic acid and the like; a lubricant such as magnesium stearate; and, an agent such as sucrose, lactose or saccharin or a flavoring agent such as peppermint, oil of wintergreen, or cherry flavoring can be added. When the unit dose form is a capsule, it may contain, in addition to materials of the above type, a liquid vehicle, such as a vegetable oil or a polyethylene glycol. Other different materials may be present as coatings or in some way modify the physical form of the solid unit dose form. For example, tablets, pills or capsules can be coated with gelatin, wax, shellac or sugar and the like. A syrup or elixir may contain the active compound, sucrose as a source of enduron, methyl and propylparabens as preservatives, a colorant and a flavoring such as cherry or orange flavor. Of course, any material used to prepare some form of unit dose should be pharmaceutically acceptable and substantially non-toxic in the amounts employed. In addition, the active compound can be incorporated into preparations and sustained release devices. The active compound can also be administered intravenously or intraperitoneally by infusion or injection. The solutions of the active compound or its salts can be prepared in water, optionally mixed with a non-toxic surfactant. The dispersions can also be prepared in glycerol, polyethylene glycols, liquids, triacetin and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms. The pharmaceutical dosage forms suitable for injection or infusion may include sterile aqueous solutions or dispersions or sterile powders comprising the active ingredient which is adapted to the extemporaneous preparation of sterile injectable or sterile infusion solutions or dispersions, optionally encapsulated in liposomes. In all cases, the final dosage form must be sterile, fluid and stable under the conditions of manufacture and storage. The carrier or liquid vehicle may be a solvent or liquid dispersion medium comprising, for example, water, ethanol, a polyol (eg, glycerol, propylene glycol and polyethylene glycides and the like), vegetable oils, esters of non-toxic glycerol and suitable mixtures thereof. Proper fluidity can be maintained, for example, by the formation of liposomes, by maintaining the required particle size in the case of dispersion or by the use of surfactants. The prevention of the action of microorganisms can be carried out by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, buffer solutions of pH or sodium chloride. The prolonged adsorption of the injectable compositions can be carried out by using the compositions of agents that retard adsorption, for example, aluminum monostearate and gelatin. Sterile injectable solutions were prepared by incorporating the active compound in the required amount in the appropriate solvent with other different ingredients listed above, as required, followed by filter sterilization. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are the vacuum drying and freeze drying techniques, which give a powder of the active ingredient plus any desired additional ingredient present in the solutions sterilized by filtration previously. For topical administration, the present compounds can be administered in pure form, that is, when they are liquid. However, it will generally be convenient to administer them to the skin as compositions or formulations, in combination with a dermatologically acceptable vehicle, which may be a solid or a liquid. Useful solid carriers include finely divided solids such as talc, clay, microcrystalline cellulose, silica, alumina and the like. Useful liquid carriers include water, alcohols or glycols or water-alcohol / glycol mixtures, in which the present compounds can be dissolved or dispersed to effective levels, optionally with the aid of non-toxic surfactants. Adjuvants such as fragrances and additional antimicrobial agents can be added to optimize the properties for a certain use. The resulting liquid compositions may be applied to absorbent pads, may be used to impregnate bandages or other fabrics, or sprayed on the affected area using pump or aerosol time sprays. The liquid compositions can also be used as eye drops, mouth rinses, washes, etc. Antibacterial presaturated towels are described by Anderson (U.S. Patent No. 4,896,768). Thickeners such as synthetic polymers, fatty acids, salts of fatty acids and esters, fatty alcohols, modified celluloses or modified mineral materials can also be used with liquid carriers to form spreads, gels, ointments, soaps and the like, for application directly to the skin of the user.

Other examples of useful dermatological compositions that can be used to deliver the compounds of the formula (I) to the skin are described in Jacquet et al., (U.S. Patent No. 4,608,392), Geria (U.S. Patent No. 4,992,478), Smith and others, (U.S. Patent No. 4,559,157) and Wortzman (U.S. Patent No. 4,820,508). Useful doses of the compounds of 1 can be determined by comparing their activity in vitro and their activity in vivo in animal models, to those of an equivalent dose of camptothecin (see, for example, BC Giovanella et al., Cancer Res .. 51 , 3052 (1991)) or Hoechst 33342 (see, AY Chem et al., Cancer Res. 53, 1332 (1993)). Methods of extrapolating effective doses in mice and other animals to humans are known in the art; for example, see Patent of E.U.A. No.4,938,949. Generally, the concentration of the compounds of the formula (I) in a liquid composition, such as a lotion, will be about 0.5-25% by weight, preferably about 0.5-10% by weight. The concentration in a semi-solid or solid composition such as a gel or a powder will be about 0.1-5% by weight, preferably about 0.5-2.5% by weight. The single doses for injection, infusion or ingestion, will generally vary between 50-1500 mg and can be administered, for example, 1-3 times a day, to give levels of approximately 0.5 - 50 mg / kg, for adults.

The terbenzimidazoles present are particularly useful for treating systemic fungal infections or "deep mycosis". These infections include coccidiomycosis, chromoblastomycosis, cryptococcosis, systemic moniliasis, histoplasmosis, aspergillosis, rodotorulosis, sporotrichosis, paracoccidiosis, phycomycosis, blastomycosis, and candidiasis. Susceptible fungi include candida (monilia) albicans, which is a member of the normal flora of mucosal membranes in the respiratory, gastrointestinal, and female genital tracts. In these and other places they may have dominance and be associated with pathological conditions. Sometimes it produces progressive systemic disease in debilitated or immunosuppressed patients. Candida can cause infection in the bloodstream, thrombophlebitis, endocarditis or infection in the eyes and other organs when it is introduced intravenously (tubes, needles, hyperalimentation, addition to narcotics, etc.). Other yeasts (eg, torulopsis glabrata) may be pathogenic under similar circumstances. The present compounds can also be used against infections of Cryptococcus neoformans. The fungus lives freely in the soil and is frequently found in pigeon feces. In men, it can cause primary lung infection that is occasionally followed by fatal meningitis. Blastomyces (Ajellomyces) dermatitidis infections can also be inhibited. This fungus causes a chronic granulomatous disease, North American blastomycosis, which can be limited to the skin or skin or can be extensively disseminated in the body. The present compounds can also be used against Blastomyces brasiliensis, an ascomycete that causes blastomycosis of South and Central America (paracoccidioidal granuloma), or to treat infection with H. capsulatum, which usually occurs in the respiratory tract and can lead to pneumonia cl unique and prolonged disease. Infections due to Coccidioide immitis can also be treated, which can cause illness if my child has fever, malaise, cough, headache, pain and sweating and which can lead to a highly fatal form called coccidiodal granuloma. The compounds are also effective against Geotrichum candidum, a fungus similar to yeast that produces geotrichosis, an infection of bronchi, lungs and mucosal membranes and Sporothrich (Sporotrichum) schenckii, a fungus that causes sporotrichosis, a chronic granulomatous skin infection., lymphatic tissues and others in animals and men. The present compounds can also be used to treat chromoblastomycosis, maduromycosis and phycomycosis, caused by Rhyzopus sp. o Mucor sp. The compounds present are particularly effective against the Aspergillus species. Aspergillus fumigatus and others Aspergillus sp. they have become a frequent cause of systemic fungal infection in an altered host. Patients with leukemia or lymphoma, immunosuppressed people (especially patients with S I DA or patients undergoing organ transplants) and those receiving intensive corticosteroid therapy are particularly susceptible to aspergillosis. The entrance door is the respiratory tract and in most cases there are manifestations of pulmonary aspergillosis, predominantly necrotising bronchopneumonia, hemorrhagic pulmonary infarction or granulomas (aspergillomas). The present compounds are also useful for inhibiting the growth of fungi, including yeasts on the skin of humans and animals such as domestic pets, farm animals and zoo animals. Such gram positive microorganisms include Propionibacterium acnes which is the main pathogen that causes human acne vulgaris. Fungal infections of the skin of animals and humans can also be treated, including head tub, leg tub (itching), body tub (culebri lla), pedia tub (athlete's foot), and nail tub. The fungi associated with said dermatophytoses include T. mentagrophytes, M. audevinii, T. rubrum, E. floccasum and M. pelineum. The present compounds are also effective against fungi associated with membrane infections of body cavities. These infections include, cotton on, vag initis and paronychia. See R. T. Yousef et al., Mvkosen, 21, 190 (1978) and H. Gershon, «Pharm. Sci. , 68, 82 (1979). The present compounds can also be used in cosmetic and skin-cleansing compositions such as soaps, shampoos, deodorants and skin softening lotions, where they can function as deodorants, i.e. to control bacteria that cause skin odor. . The present compounds can also be used in shampoos, rinses and other hair care products to inhibit Pityrosporum ovale (dandruff, skin lesions, in immunosuppressed subjects). Analogs present can also be used to treat cancers known to be susceptible to topoisomerase I inhibitors, including, but not limited to, Burkitt's mor, chronic lymphocytic leukemia, multiple myeloma, anaplastic squamous cell and long-cell carcinomas, adenocarcinomas of the lung , Ewing's sarcoma, non-Hodgkins's disease, breast tumor, colon tumor, stomach tumor, bronchogenic carcinoma of oat cells, squamous cell carcinoma of the cervix, ovarian tumors, bladder tumors, testicular tumors , endometrial tumors, malignant melanoma and lymphocytic leukemia, water and prostatic carcinoma. The present compounds can be administered as agents alone or in combination with other antineoplastic drugs commonly used to treat these cancers. The invention will also be described by reference to the following detailed examples wherein the melting points were determined with a Thomas-Hoover melting point single melting capillary apparatus. The infrared spectrum data (I R) were obtained in a Fourier transformation spectrophotometer 1600 from Perkin-Elmer and reported in cm'1. Nuclear magnetic resonance of protons (1H NMR) and carbon (13C NMR) were recorded in a Fourier transformation spectrophotometer of Varian Gemini-200. The NMR spectra (200 MHz 1H and 50 MHz 13C) were recorded in CDCI3 (unless otherwise noted) with chemical changes reported in units d lower than tetramethylsilane (TMS). Coupling constants were reported in hertz. The mass spectra were obtained from the Midwest Center for Mass Spectrometry within the Department of Chemistry of the University of Nebraska-Lincoln. The combustion analyzes were carried out by Atlantic Microlabs, Inc., Norcross, GA, and were at + 0.4%. THF was recently distilled from sodium and benzophenone before use. Allytributyltin and phenyltributyltin were purchased from Aldrich Chemical Company. Strain R21 of Aspergillus nidulans (pabaAl, and A2) was used in all the examples. Hoechst dye of dibenzimidazole 33342 (Ho33342), camptothecin and berenyl dye, were compared to Sigma Chemical Co. Monobenzimidazoles (QS / ll / 9, 48, 50, 51 and 59A), terbenzimidazoles (H and 1_3) and protoberberines (coral, DMII / 33) and nitidine were synthesized as described below and by (Q. Sun et al., Biorg. &; Med. Chem. Lett. 4, 2871 (1994) and J. Med. Chem., 38, 3638 (1995); Kim and others, Biorg. & Med. Chem. Lett. 462 (1996); J. Med. Chem .. 39, 992 (1996); D. Makhey et al., Med. Chem. Res .. 5, 1 (1995); Biorg. & Med. Chem. Lett. 4, 781 (1996)). (See Figure 8 for structures). All drugs were dissolved in dimethyl sulfoxide (Sigma Chemical Co.) at a concentration of 1, 5 or 10 mg / ml and kept frozen in aliquots at -20 ° C. Example 1. General Procedure for Coupling Reaction Catalyzed by PdCUfPPha)? of 4-Bromo-2-nitroaniline (13) with Tin Compounds (A) 4-Phenyl-2-nitroaniline (5). A solution of 4-bromo-nitroaniline 17 (1.0 g, 4.67 mmol), tributylphenyl tin (2.2 g, 6.07 mmol), bis (triphenylphosphine) palladium (II) chloride (164 mg, 0.234 mmol) and triphenylphosphine (613 mg, 2.34 mmole) in DMF (15 ml) was heated under N2 at 120 ° C overnight. After the solution was cooled to room temperature, the reaction mixture was chromatographed directly on silica gel with 2-5% EtOAc / Hexane to give 752 mg (75) of 5 as a yellow solid: mp 169-171 ° C; IR (CHCl3) 3517, 3398, 3022, 1635, 1525, 1250; 1H NMR 8.38 (1H, d, J = 2.2), 7.66 (1H, dd, J = 8.7, 2.2), 7.59-7.54 (2H, m), 7.49-7.34 (3H, m), 6.90 (1H, d , J = 8.8), 6.13 (NH, brs); 13 C NMR 144.2, 139.3, 135.0, 130.9, 129.5, 127.8, 126.8, 124.4, 119.8, 112.8; Anal. Cale, for C? 2H10N2O2: C, 67.28; H, 4.70; N, 13.08. Found: c, 67.38, H, 4.76; N, 13.01. (B) 4-AMI-2-nitroaniline (4). Prepared from 4-bromo-2-nitroaniline 17 (1.70 g, 7.84 mmol) and allyltributyltin (3.38 g, 10.2 mmol) as a yellow solid with 96% yield as described above for 5: mp 29-31 ° C; IR (KBr) 3490, 3374, 1638, 1518, 1341, 1253; 1H NMR 7.90 (1H, d, J = 2.0), 7.19 (1H, dd, J = 8.5, 2.0), 6.77 (1H, d, J = 8.5), 6.05 (NH, brs), 6.00-5.80 (1H , m), 5.11 (1H, dd, J = .4, 1.4), 5.04 (1H, ddd, J = 6.6, 3.0, 1.5), 3.28 (1H, d, J = 6.6); 13 C NMR 143.81, 137.13, 129.34, 125.59, 119.49, 116.95, 39.18; HRMS (El) Cale. For C9H10N2O2 178.0742, found 178.0746. (C) 4- (2'-Pyridyl) -2-nitroaniline (6). Prepared from 4-bromo-2-nitroaniline 17 (597 mg, 2.75 mmol) and 2-tributylstanilpyridine (1.01 g, 2.75 mmol) as a yellow solid with 52% yield as described above for 5: mp 146-148 ° C; IR (CHCl3) 3516, 3397, 3020, 1634, 1524, 1341, 1250; 1H NMR 8.74 (1H, d, J = 2.2), 8.63 (1H, dd, J = 4.9, 1.5), 8.13 (1H, dd, J = 8.8, 2.1), 7.78-7.66 (2H, m), 7.20 (1H, ddd, J = 4.8, 4.7, 1.9), 6.92 (1H, d, J = 8.8), 6.37 (NH, brs); 13 C NMR 155.6, 150.1, 145.6, 137.4, 134.5, 129.1, 124.7, 122.4, 119.8, 119.7; Anal. Cale. For CnH9N3O2: C, 61.39; H, 4.21; N, 19.53. Found: C, 61.29; H, 4.23; N, 19.43. (D) 4- (3'-Pyridyl) -2-nitroaniline (7). Prepared from 4-bromo-2-nitroaniline 17 (1.42 g, 6.53 mmol) and 3-tributylstanilpyridine (3.60 g, 9.79 mmol) as a yellow solid with 32% yield as described above for 5: mp 177-179 ° C; IR (CHCl3) 3515, 3399, 3052, 2983, 1638, 1524, 1341, 1259; 1H NMR 8.68 (1H, dJ = 1.7), 8.42 (1H, dd, J = 4.8, 1.5), 8.22 (1H, d, J = 2.2), 7.74 (1H, ddd, J = 7.9, 2.4, 1.6) , 7.50 (1H, dd, J = 8.7, 2.2), 7.23 (1H, ddd, J = 8.0, 4.8, 0.8), 6.92 (1H, d, J = 8.8), 6.56 (NH, brs); 13 C NMR 148.7, 147.8, 145.4, 135.0, 134.4, 133.8, 126.5, 124.4, 124.0, 120.4; Anal. Cale. For CnH9N3O2: C, 61.39; H, 4.21; N, 19.53. Found: C, 61.28; H, 4.16; N, 19.40. (E) 4- (4'-Pyridyl) -2-nitroaniline (8). Prepared from 4-bromo-2-nitroaniline 17 (165 mg, 0.76 mmol) and 4-tributylstanilpyridine (280 mg, 0.76 mmol) as a yellow solid in 25% yield as described above for 5: mp 230-232 ° C; IR (CHCl3) 3518, 3398, 3032, 1636, 1528, 1344; 1H NMR 8.55 (2H, d, J = 6.3), 8.52 (1H, d, J = 2.3), 7.84 (1H, dd, J = 8.9, 2.3), 7.71 (2H, d, J = 6.4), 7.13 (1H, d, J = 8.9); 13 C NMR 149.4, 133.4, 124.0, 120.7, 120.0; HRMS (El) Cale, for CnH9N3O2 215.0695. Found 215.0698. Example 2.5-Form i l-2- (benzimidazol-5'-yl) benz imidazole (9). A mixture of 5-cyano-2- (benzimidazol-5'-yl) benzimidazole 19 (148 mg, 0.57 mmole), Ni-AI catalyst (500 mg), formic acid (7 ml) and water (3 ml), was heated under reflux under N2 for 4 hours. The hot reaction mixture was filtered immediately through a plug of celite and evaporated to give a yellow solid. The yellow solid was then dissolved in hot water (5 ml) and the solution was neutralized to pH 9 by 2N NaOH. The solid precipitate was recovered by suction filtration and further purified by flash chromatography on silica gel (15% MeOH / EtOAc) to give 142 mg (95%) of 9 as a white solid: mp >275 ° C; IR (KBr) 3106, 2835, 1685, 1618, 1432, 1293; 1H-NMR (CD3OD) d 10.01 (1H, s), 8.39 (1H, s), 8.35 (1H, s), 8.13 (1H, s) 8.06 (1H, dd, J = 8.6, 1.6), 7.83 (1H, s), dd, J = 8.4, 1.4), 7.77 (1H, d, J = 8.5), 7.71 (1H, d, J = 8.3); HRMS (FAB) Cale, for C? SHnN4O 263.0933, found 263.0932. Example 1. General Procedures for Preparing 5- Substituted Trisbenzimidazoles (A) 2- [2 '- (Benzimidazol-5"-i I) benzimidazol-5'-yl] benzimidazole (10) A solution of 5-formyl-2- (benzimidazol-5'-yl) benzimidazole 9 (121 mg, 0.46 mmol) and phenylenediamine (60 mg, 0.55 mol) in nitrobenzene (8 mL) was heated at 150 ° C under N2 for the night. The mixture was cooled to room temperature and chromatographed on silica gel (0.20% MeOH / EtOAc) to give 155 mg (96%) of 10 as a yellow solid: mp > 275 ° C; IR (KBr) 3400, 3157, 1630, 1542, 1438, 1294; 1 H NMR (DMSO-d 6 + 3 drops of CF 3 COOH) d 9.71 (1H, s), 8.75 (1H, s), 8.65 (1H, d, J = 1.1), 8.48 (1H, dd, J = 8.7, 1.5) , 8.21 (1H, dd, J = 8.6, 1.6), 8.15 (1H, d, J = 8.8), 8.08 (1H, d, J = 8.7), 7.90 (1H, dd, J = 6.2, 3.1), 7.61 (2H, dd, J = 6.1, 3.1); 13 C NMR (DMSO-d 6 + 3 drops of CF 3 COOH) d 154.4, 149.8, 133.2, 132.0, 131.7, 126.2, 125.5, 125.4, 123.9, 123.6, 116.3, 115.9, 114.23, 114.17, 114.13; HRMS (FAB) Cale, for C2? H? SN6351.1358, found: 351.1357. (B) 5-Cyano-2- [2 '- (Benzimidazol-5"-yl) benzimidazol-5'-yl-benzimidazole (11). Hydrogenation of 3 (70 mg, 0.43 mmol) was achieved at 2812 kg / cm2. H2 at room temperature for 1 hour using 10% Pd-C (30 mg) in EtOAc (10 mL) The reaction mixture was filtered and concentrated in vacuo to give a solid.The solution of this solid and 9 (87 mg, 0.33 mmol) in nitrobenzene (5 mL) was heated at 150 ° C under N2 overnight.The mixture was cooled to room temperature and chromatographed directly on silica gel (0-10% MeOH / EtOAc) to give 107 mg (86%) of 11 as a solid, mp> 280 ° C; IR (KBr) 3416, 3148, 2222, 1626, 1553, 1441, 1291; 1H-NMR (DMSO-d6 + 3 drops of CF3COOH) d 8.50 (1H, s), 8.46 (1H, s), 8.40 (1H, s), 8.18-8.11 (3H,), 7.81-7.75 (3H, m), 7.62 (1H, dd, J = 8.3, 1.5); HRMS (FAB) Cale, for C22H13N7 376.1310, found: 376.1309. (C) Propyl-2- [2 '- (benzimidazol-5"-yl) benzimidazol-5'-yl] benz-midazole (12). Prepared 4-Allyl-2-nitroaniline 4 (312 mg, 1.75 mmol) and 5-formyl-2- (benzimidazol-5'-yl) benzimidazole 9 (121 mg, 0.46 mmol) with a yield of 79% as described before for 11: solid: pf > 270 ° C; IR (KBr) 3421, 3068, 2957, 1434; 1 H NMR (DMSO-d 6 + 3 drops of CF 3 COOH) d 9.66 (1H, s), 8.73 (1H, s), 8.59 (1H, s), 8.48 (1H, dd, J = 8.7, 1.5), 8.13 (1H , dd, J = 8.7, 1.4), 8.11 (1H, d, J = 8.7), 8.02 (1H, d, J = 8.5), 7.79 (1H, d, J = 8.4), 7.66 (1H, s); 7.45 (1H, dd, J = 8.5, 1.3), 2.80 (2H, t, J = 7.0), 1.70 (2H, m), 0.96 (3H, t, J = 7.2); 13 C NMR (DMSO-d 6 + 3 drops of CF 3 COOH) d 153.84, 149.74, 141.64, 141.01, 139.37, 133.10, 132.26, 131.99, 130.34, 127.08, 126.26, 125.14, 141.64, 141.01, 139.37, 133.10, 132.26, 131.99, 130.34 , 127.08, 126.26, 125.14, 122.91, 117.52, 116.32, 116.06, 115.76, 113.78, 112.99, 37.45, 24.73, 13.74. (D) 5-Phenyl-2- [2 '- (benzimidazol-5"-yl) benzimidazol-5'-yl-benzimidazole (13) .Preparation of 4-phenyl-2-nitroaniline 5 (247 mg, 1.15 mmoles) and -formyl-2- (benzimidazol-5'-yl) benzimidazole 9 (201 mg, 0.77 mmol) with a yield of 89% as described for 11: solid: mp 262-164 ° C dec; IR (KBr) 3402, 3104, 1627, 1552, 1442, 1290; 1H-NMR (DMSO-d6 + 3 drops of CF3COOH) d 9.66 (1H, s), 8.74 (1H, s), 8.65 (1H, s), 8.50 (1H, dd, J = 8.8, 1.1), 8.21 (1H, dd, J = 8.7, 1.4), 8.12 (1H, d, J = 8.8), 8.06 (1H, s), 8.05 (1H, d, J = 8.4), 7.97 (1H, d, J = 8.7), 7.89 (1H, dd, J = 8.7, 1.5), 7.80 (2H, d, J = 7.0), 7.61-7.47 (3H, m); HRMS (FAB) Cale, for C27H19N6 427.1671, found 427.1666. (E) 5- (2-Pyridyl) -2- [2 '- (benzimidazol-5"-yl) benzimidazol-5'-yl-benzimidazole (14). Preparation of 4- (2'-pyridyl) -2-nitroaniline 6 (110 mg 0.60 mmol) and 5-formyl-2- (benzimidazol-5'-yl) benzimidazole 9 (51 mg, 0.25 mmol) with a yield of 84 % as described above for 11: solid: pf > 275 ° C; IR (KBr) 3411, 3157, 1630, 1593, 1432; 1H-NMR (CD3OD) d 8.59 (1H, d, J = 4.8), 8.35 (1H, s), 8.31-8.25 (2H, m), 8.10 (1H, s), 8.04-7.94 (2H, m), 7.85 -7.77 (3H, m), 7.72 (1h; d, J = 8.6), 7.68 (1H, d, J = 8.7), 7.64 (1H, d, J = 8.7), 7.30 (1H, m); HRMS (FAB) Cale, for C26H18N7428.1624, found 428.1611. (F) 5- (3-Pyridyl) -2- [2 '- (benzimidazol-5"-yl) benzimidazol-5'-yl-benzimidazole (15) .Preparation of 4- (3'-pyridyl) -2-nitroaniline 7 (183 mg, 0.85 mmol) and 5-formyl-2- (benzimidazol-5'-yl) benzimidazole 9 in 46% yield as described above for 11: solid: mp >275 ° C; IR (KBr) 3400, 3070, 2836, 1438, 1289; 1H-NMR (CD3OD) d 8.83 (1H, d, J = 1.6), 8.49 (1H, dd, J = 4.9, 1.5), 8.38 (1H, d, J = 1.1), 8.31 (1H, d, J = 1.1 ), 8.29 (1H, s), 8.11 (1H, ddd, J = 8.0, 2.3, 1.6), 8.05 (1H, dd, J = 8.5, 1.6), 8.0 (1H, dd, J = 8.5, 1.6), 7.81 (1H, d, J = 1.1), 7.77-7.68 (3H, m), 7.55-7.47 (2H, m); HRMS (FAB) Cale, for C26H18N7428.1624, found 428.1612. (G) 5- (4-Pyridyl) -2- [2 '- (benzimidazol-5"-yl) benzimidazole-5'-Ijbenzimidazole (16) .Preparation of 4- (4'-pyridyl) -2-nitroaniline 8 (35 mg, 0.16 mmol) and 5-formyl-2- (benzimidazol-5'-yl) benzimidazole 9 (50 mg, 0.19 mmol) with a 43% yield as described above for 11: solid: mp > 280 ° C; IR (KBr) 3411, 3118, 1600, 1552, 1439, 1290; 1H-NMR (CD3OD) d 8.51 (2H, d, J = 6.2), 8.33 (1H, d, J = 1.1), 8.27 ( 1H, s), 8.25 (1H, d, J = 1.1), 8.01 (1H, dd, J = 8.6, 1.7), 7.96 (1H, dd, J = 8.9, 2.0), 7.87 (1H, d, J = 1.0), 7.74-7.56 (6H, m); HRMS (FAB) Cale, for C26H18N7428.1624, found 428.1625. Example 4.4-Bromo-2-nitroaniline (17) A solution of 2-nitroaniline (5 g, 36.2 mmoles) ) in CH2CI2 (100 mL) was cooled to -10 ° C and treated with 90% 2,4,4,6-tetrabromo-2,5-cyclohexadienone (19.8 g, 43.5 mmol) in 5 portions. The mixture was stirred at -10 ° C-0 ° C for 1 hour. After warming to room temperature, the reaction mixture was washed with 2N NaOH (60 ml) and brine (50 ml), dried over Na2SO4 and evaporated. Flash chromatography on silica gel (5% EtOAc / Hexane) gave 7.40 g (94% of 17 as a yellow solid: mp 109-110 ° C (lit. mp 112-113 ° C); 1 H NMR 8.27 ( 1H, d, J = 2.3), 7.43 (1H, dd, J = 8.9, 2.4), 6.73 (1H, d, J = 8.8), 6.09 (NH, brs).

Example 5.5-Formylbenzimidazole (18). A suspension of 5-benzimidazolecarboxylic acid (1.57 g, 9.7 mmol) in dry THF (50 ml) was cooled to -78 ° C under N2 and treated with LiAIH4 (736 mg, 19.14 mmol). After the addition, the mixture was allowed to warm slowly to room temperature and then stirred at room temperature overnight. The mixture was cooled by MeOH and H2O carefully and passed through a short column of silica gel eluting with 10% MeOH / EtOAc. The eluate was concentrated to give 876 mg of crude alcohol as a solid. The crude alcohol (876 mg) was dissolved in a mixture of DMF (3 ml), THF (10 ml) and CH2Cl2 (40 ml). The 4-methylmorpholine N-oxide (2.25 g, 19.2 mmoles), molecular sieves of 4Á (5 g) and TPAP (169 mg, 0.48 mmole) were subsequently added to the crude alcohol solution. The mixture was stirred at room temperature overnight and filtered through a pad of silica gel eluting with 0-10% MeOH / EtOAc to give 452 mg (32%, 2 steps) of 17 as a white solid: mp 164-166 ° C IR (KBr) 3087, 2818, 1690, 1292; 1H-NMR (CD3OD) d 9.95 (1H, s), 8.34 (1H, s), 8.08 (1H, d, J = 1.5), 7.74 (1H, dd, J = 8.4, 1.5), 7.63 (1H, d, J = 8.4); 13 C NMR (CD3OD) d 194.2, 146.0, 143.0, 139.8, 133.6, 124.9, 120.7, 116.6: Anal. Cale, for C8H6N2O: C, 65.75; H, 4.14; N, 19.17. Found: C, 65.60; H, 4.17; N, 19.08. Example 6.5-Cyano-2- (benzimidazol-5'-yl) benzimidazole (19). A mixture of 5-formylbenzimidazole 18 (211 mg, 1.44 mmol) and 4-cyano-1,2-phenylenediamine (230 mg, 1.73 mmol) in nitrobenzene (10 ml) was heated at 150 ° C under N2 overnight. The mixture was cooled to room temperature and chromatographed directly on silica gel eluting with 0-15% MeOH / EtOAc to give 244 mg (65%) of 18 as a solid: mp > 270 ° C IR (KBr) 3110, 2826, 2224, 1627, 1426, 1294; 1H-NMR (CD3OD) d 8.41 (1H, s), 8.33 (1H, s), 8.07 (1H, dd, J = 8.6, 1.5), 7.98 (1H, s), 7.78 (1H, d, J = 8.4) , 7.73 (1H, d, J = 8.4), 7.56 (1H, dd, J = 8.4, 1.5), 13C NMR (DMSO-d6 + 3 drops of CF3COOH) d 153.4, 140.4, 138.3, 132.9, 131.6, 127.0, 125.8, 125.3, 120.8, 119.8, 116.0, 115.8, 113.9, 105.5; HRMS (FAB) Cale, for Ci5H10N5260.0936, found 260.0935. Example 7 (A) 5-Bromo-2- [2 '- (benzimidazol-5"-l) benzimidazol-5'-yl] -benzimidazole (JSKI IV-37) A mixture of 5-formyl-2- (benzimidazole) -5'-yl) benzimidazole (118.8 mg, 0.45 mmol) and 5-bromophenylenediamine (169.6 g, 0.9 mmol) in nitrobenzene (5 mL) was heated at 150 ° C under N2 overnight.The mixture was cooled to room temperature and chromatographed using 0-10% methanol / ethyl acetate to give 127.3 mg (66%) of brown-yellow solid: mp> 280 ° C IR (KBr) 3101, 1626, 1547, 1440; 1 H NMR (DMSO) d6) d 7.34 (dd, 1H, J = 7.0, 2.0), 7.57 (d, 1H, J = 9.0), 7.71-7.80 (m, 3H), 8.04-8.18 (m, 2H), 8.39 (s, 2H) ), 8.50 (s, 1H); 13C NMR (DMSO-d6 + 3 drops of CF3COOH) d 114.1, 115.8, 116.2, 116.4, 117.4, 117.0, 118.6, 123.5, 125.3, 126.2, 128.7, 128.9, 131.8, 132.0, 132.3, 133.1, 134.4, 138.3, 140.6, 151.1, 153.4.

(B) 5-Chloro-2- [2 '- (benzimidazol-5"-yl) benzimidazol-5'-i!] - benzimidazole (JSKI IV-68) A mixture of 5-formyl-2- (benzimidazole-5) '-il) benzimidazole (160 mg, 0.61 mmol) and 5-chlorophenylenediamine (174 mg, 1.22 mmol) in nitrobenzene (5 mL) was heated at 150 ° C under N2 overnight.The mixture was cooled to room temperature and chromatographed using 0-10% methanol / ethyl acetate to give 167 mg (71%) of brown-yellow solid: mp> 280 ° C IR (KBr) 3103, 2826, 1427, 1293; 1 H NMR (DMSO-d6) d 7.24 (dd, 1H, J = 8.5, 2.0), 7.60-7.81 (m, 4H), 8.07-8.17 (m, 2H), 8.40 (s, 2H), 8.50 (s, 1H); 13C NMR (DMSO -d6 + 3 drops of CF3COOH) d 114.3, 114.4, 115. 3, 115.5, 115.6, 116.2, 118.5, 123.1, 125.4, 125.5, 125.6, 129.4, 132. 4, 132.9, 133.0, 135.2, 138.9, 140.9, 151.8, 153.5. (C) 5- (p-Chlorofinyl) -2- [2 '- (benzimidazol-5"-yl) benzimidazol-5'-yl] -benzimidazole (JSKI IV-47) A mixture of 5-formyl-2 - (benzimidazol-5'-yl) benzimidazole (99 mg, 0.38 mmol) and 5- (p-chlorophenyl) phenylenediamine (154 mg, 0.71 mmol) in nitrobenzene (5 mL) was heated at 150 ° C under N2 overnight The mixture was cooled to room temperature and chromatographed using 0-10% methanol / ethyl acetate to give 85 mg (49%) of brown yellow solid: mp> 280 ° C IR (KBr) 3046, 2820, 1426 1281; 1H NMR (DMSO-d6 + 3 drops of CF3COOH) d 7.56 (d, 2H, J = 8.5), 7.82 (d, 2H, J = 8.5), 7.88-8.21 (m, 6H), 8.48 (d , 1H, J = 8.8), 8.63 (s, 1H), 8.72 (s, 1H), 9.69 (s, 1H); 13C NMR (DMSO-d6 + 3 drops of CF3COOH) d 118.8, 114.7, 115.8, 116.1, 117.7, 123.0, 124.1, 125.2, 125.3, 129.2, 129.3, 131.9, 132.1, 133.0, 133.1, 137.2, 138.5, 139.3, 141.6, 150.8, 153.8. (D) 4-Bromophenylenediamine (JSK IV-35) A 2 -nitro-4-bromoaniline (340 mg, 1.57 mmol) in absolute ethanol (20 mL) was added SnC I2 (1.50 g, 7.91 mmol) and heated to reflux overnight. The reaction mixture was then basified to pH 11 with 2N NaOH and extracted with ether to give 275 mg (94%) of product. This product was used without further purification for the synthesis of JSK IV-37. (E) 4-Chlorophenylenediamine (JSK IV-67) To 2-nitro-4-chloroaniline (304 mg, 1.76 mmol) in absolute ethanol (20 mL) was added SnCl2 (1.68 g, 8.86 mmol) and heated to reflux overnight. The reaction mixture was then basified to pH 11 with 2N NaOH and extracted with ether to give 250 mg (quantitative yield) of product. This product was used without further purification for the synthesis of JSK IV-68. (F) p-Chlorotributyl phenyltin (JSK IV-42) 4-Bromochlorobenzene (3.2 g, 16.62 mmol) was dissolved in dry THF (20 ml). After bringing the reaction temperature to -78 ° C with an acetone / dry ice bath, nBuLi (15.58 mL, 1.6M, 1.5 equivalent) was added slowly and stirred at -78 ° C for 30 minutes. Tributyl tin chloride (6.77 mL, 1.5 equivalent) was added and stirred overnight while the reaction was brought to room temperature. The reaction mixture was cooled by stirring the open reaction flask in air for 1 hour after which TH F (7.35 G, 97%) was rotoevaporated, then passing the mixture through a column of rapid silica gel with 100% strength. hexanes.

(G) 2-Nitro-5- (p-chlorophenyl) aniline (JSK IV-44) To JSK IV-42 (2.02 g, 5.04 mmole) and 2-nitro-4-bromoaniline (730 mg, 3.36 mmole) in DMF (18 μL) was added Pd (PPh3) 2Cl2 (17.9 mg, 0.17 mmol) and PPh3 (440.2 mg, 1.70 mmol) and heated at 120 ° C overnight. DM F was rotoevaporated and the mixture was separated on a column of silica gel eluting with 5-10% ethyl acetate / hexanes to give 270 mg (32%) reddish solid. (H) 4- (p-Chlorophenyl) phenylenediamine (JSK IV-46) JSK was dissolved IV-44 (190 mg, 0.77 mmol) in ethyl acetate (100 μL) and then adding 10% Pd-C (40 mg) was reduced by hydrogenation (3.1635 kg / cm2). The product (quantitative yield) was used in JSK IV-47 without further purification. Example 8. Bioanalysis A. Analysis of DNA Separation Mediated by Topoisomerase I Topoisomerase I of A DN was purified from calf thymus gland as previously reported by BD H illigan et al., J_ ^ Biol. Chem. 260. 2475 (1985). Plasmid YEpG was also purified by the alkali lysis method followed by phenol deprotection and isspecific centrifugation of CsCI / etidinium as described by T. Mariatis et al.

Molecular Cloning, A Laboratorv Manual, Cold Spring Harbor Labs, NY (1982) pages 149-185. The final labeling of the plasmid was achieved as previously described by L.F. Liu et al., J. Biol. Cherr.: T 258, 15365 (1983). Separation analyzes were carried out as previously reported by A. Y. Chem et al., Cancer Res., 53, 1332 (1993). Human topoisomerase I was isolated as a recombinant fusion protein using the T7 expression system. B. Cytotoxicity Analysis Cytotoxicity was determined using the tetrazolium cytotoxicity assay of the MTT microtiter plate (MTA) following the procedures of F. Denizot et al., J. Immunol. Methods, 89, 271 (1986); J. Carmichael et al., Cancer Res., 47, 936 (1987) and T.J. Mosmann et al., Immunol. Methods. 65. 55 (1983). The human lymphoblast RPMI 8402 and its camptothecin-resistant variant cell line CPT-K5, were provided by Dr. Toshiwo Andoh (Aichi Cancer Center Research Institute, Nagoya, Japan). See, for example, T. Andoh et al., Adv. Pharmacol., 29B, 93 (1994). The cytotoxicity analysis was carried out using 96-well microtiter plates. The cells were grown in suspension at 37 ° C in 5% CO2 and a regular passage was maintained in RPMI medium supplemented with 10% heat inactive fetal bovine serum, L-glutamine (2 mM), penicillin (100 U / ml) and streptomycin (0.1 mg / ml). For the determination of IC5o the cells were exposed continuously with varying concentrations of drug concentrations and MTT analysis was carried out at the end of the fourth day. The KB3-1 cell line of human drug-sensitive squamous cell carcinoma (S. Auyama et al., Somatic Cell Mol. Genet., 11, 117 (1985)) and its cells of KBV-1 multidrug-resistant variant selected from vinblastine (DW Shen et al., Science, 32, 643 81986) were provided by Dr. Michael Gottesman (National Cancer Institute, Bethesda, ML). These cells were grown as single-layer cultures in 5% CO2 and maintained in a regular passage in minimal essential Dulbecco medium supplemented with 10% heat-inactivated fetal bovine serum. The KBV-1 cells maintained similarly except that they were grown in the presence of 1 μg / ml of vinblastine. C. Results As shown in Table 1, the comparison of compounds 10-16 and halo analogues of JSKIB-37, 47 and 68 with Hoechst 33342 (1) as topoisomerase I inhibitors, demonstrated that several of these trisbenzimidazoles had potency. similar or greater Table 1. Separation of DNA mediated by topoisomerase I and Cytotoxicity of Bis and Trisbenzimidazoles Separation of Cytotoxicity of ICsoa (μM) DNA mediated Cell Lines Compound by Topo I RPMI CPT-K5 KB3-1 KB-1 Hoechst 33342 1 0.03 0.9 0.01 1.2 1.1 14 28 N.D. N.D. 11 1 > 25c > 25c N.D. N.D. 12 100 7.9 20 N.D. N.D. 13 2 0.09 0.58 0.58 0.35 14 3.3 0.16 5.8 0.05 0.09 2 0.035 2.5 0.02 0.02 16 2 0.035 2.5 0.02 0.01 19 1000 > 25c N.D N.D. N.D.

JSKIV-37 1 1.40 1.40 JSKIV-47 10 0.09 0.20 JSKIV-68 1 1.04 0.65 a) IC50 was calculated after 4 days of continuous exposure to the drug, N.D. = Not determined. b) Topoisomerase I separation values were reported as CER, Relative Effective Concentration, that is, the relative concentrations of Hoechst 33342, whose value was assumed arbitrarily as 1, which are capable of producing the same separation in the plasmid DNA in presence of topoisomerase I of the calf thymus. The separation was calculated from the strongest intensity of the specific Hoechst band. c) No indication of cytotoxicity was considered indicative of IC50 values substantially greater than the upper doses analyzed. While 10 and 11 exhibited similar potency in their inhibition of topoisomerase I as observed with Hoechst 33342, both compounds did not exhibit significant cytotoxicity towards the human lymphoblast cell line, RPMI 8402. However, this may be due to the inability of the compound pure penetration of the target cell that can be overcome by the selection of a suitable vehicle, such as liposomes. Trisbenzimidazole substituted with 5-phenyl 13 was about half as potent as Hoechst 33342 as a topoisomerase I inhibitor. However, in contrast to 10 and 11, it had significant cytotoxicity towards the human lymphoblast cell line, RPMI 8402 cells. As observed with Hoechst 33342, 13 was also effective against Camptothecin-resistant CPT-K5 cells. The relative resistance of Hoechst 33342 and 13, were expressed as the ratio of IC50 values of the resistant cell line to the drug sensitive one, is approximately 30 times compared to the relative resistance of camptothecin which is 2,500 times, as reported by AY Chem et al., Cancer Res., 53, 1332 (1993). A similar effect was observed in another pair of cell lines; 13 has an IC50 of 0.015 μg / ml in the human ovarian tumor cell line, A2780, relative to an IC50 of 0.03 μg / ml in CPT-2000, a variant of A2780 selected for camptothecin resistance and known to contain a topoisomerase I resistant to mutant camptothecin. The trisbenzimidazole derivative of 5-n-propyl, 12, was much less active than 10, 11 or 13 as a topoisomerase I inhibitor. Its weak activity as a topoisomerase I inhibitor correlated with its weak cytotoxicity. The activity of several of these compounds was also evaluated using recombinant human topoisomerase I. Several of these analogs induced separation of DNA in the presence of human topoisomerase I compared to that observed with calf-type isolated topoisomerase I. The cytotoxicity activity of Hoechst 33342 and 13 was also evaluated against KB-31 and KB V-1 cells. The primary difference between these cell lines is in the degree to which human MDR1 is expressed (P-glycoprotein). Recent studies have shown that antineoplastic agents that are cationic at physiological pH probably serve more as substrates for MDR1 and, therefore, are similarly less active against cells that secrete P-glycoprotein. In view of the fact that Hoechst 33342 is extensively protonated at physiological pH, it is not surprising that ICso differs from approximately two orders of magnitude for KB 3-1 compared with KB V-1 cells, as reported by AY Chem et al., Adv. . Pharmacol .. 245. 29B (1994). In contrast to Hoechst 33342, there is little difference between the ICS0 values observed for 13 in these two cell lines. Therefore, 13 does not appear to be a substrate for human MDR1. These data indicate that these trisbenzimidazole derivatives have significant chemotherapeutic advantages purchased with Hoechst 33342 or pibencimol (Hoechst 33258)., 2 '- (4-hydroxyphenyl) -5- (4-methyl-1-piperazinyl) -2,5'-bi-1H-benzimidazole. These data indicate that the substitution of this trisbenzimidazole with the 5-Ar substituent can give derivatives that are active as topoisomerase I inhibitors and cytotoxic for tumor cells. The trisbenzimidazoles substituted at the 5-position with the 2-, 3-, or 4-pyridyl group. 14-16, were evaluated for their potency as inhibitors of topoisomerase I and for cytotoxicity as summarized in Table 1. These analogues, similar to 13, have activity as inhibitors of topoisomerase I. The 3- and 4-pyridyl analogues, 15 and 16, are somewhat more active than the 2-pyridyl derivative, 14, as the topoisomerase I inhibitors as well as the cytotoxic agents. As can be seen with 13, these trisbenzimidazoles substituted with pyridyl had cell-like cytotoxicity KB 3-1 as well as KB V-1 cells that overexpress M DR 1. A primary advantage of these trisbenzim idazoles substituted with heteroaryl compared to Hoechst 33342 is their efficacy against cell lines expressing MDR 1. Example 9. Partial Purification of Topoisomerase I from Asoeraillus nidulans. Two liters of YG medium (0.5% yeast extract and 2% glucose) were inoculated with approximately 5 x 108 conidia / ml. After 16 hours of development at 37 ° C, the mycelia were recovered, washed with p HI buffer (50 mM Tris-HCl, pH 7.7, 1 mM EDTA, 1 mM EGTA, 10% glycerol, 1 mM of phenylmethylsulfonyl fluoride and 1 mM of 2-mercaptoethanol) and rapidly cooled in nitrogen. Frozen mycelia (approximately 20 grams) were milled to powder and resuspended in 30 ml of Regulatory Solution I. The lysate was centrifuged at 10 K rpm on a Rotor Sorval H B3 for 15 minutes to remove cellular debris. The supernatant was made at 6% in polyethylene glycol (v / v) and 1 M NaCl. After one hour on ice with moderate agitation, the solution was centrifuged at 14 K rpm in a Rotor Sorball for 30 minutes to remove the nucleic acids. The subsequent purification steps were the same as those previously described by purification of topoisomerase from recombinant human DNA (B. Gatto et al., Cancer Res. 56, 2795 (1996)). Briefly, the supernatant was directly chromatographed on an HPLC column of Hydroxyapatite Bio-gel (BioRad Laboratories, Richmond, CA). Fractions containing the relax activity were combined, diluted and then loaded onto a BioRex70 column (BioRad Laboratories, Richmond, CA). The column was revealed with a linear gradient of 0.2 to 1 M KCI. The peak fractions were combined and dialysed overnight at 4 ° C against 30 mM potassium phosphate, 50% glycerol (v / v), 0.5 mM EDTA, and 1 mM DTT. Recombinant human topoisomerase I was purified from Eschericha coli BL21 (DE3) harboring PET1B as previously described by (Gatto et al., Cancer Res..56, 2795 (1996)). Example 10. Covalent Transfer of 32P Radioactivity from DNA to Topoisomerase I. The phosphate transfer method was a modification of the procedure previously described by T.C. Rowe et al., J. Biol. Chem., 259. 9177 (1984). In summary, a 100 μl reaction mixture containing 10 mM Tris-HCl, pH 7.5, 1 mM MgCl 2, 0.5 mM dithiothreitol, 30 μg / ml bovine serum albumin, drug (camptothecin or Hoechst 33342) at an indicated concentration, 50 ng of YEpG, labeled with 32PdATP DNA by the random starter method (Randomly Initiated Marking Equipment, Boehringer Mannheim) and 300 units of human topoisomerase I or Aspergillus, was incubated at 37 ° C for 10 minutes . The reactions were terminated by adding 0.18 M NaOH and 2.5 mM EDTA. After neutralizing the reaction with a pre-calibrated amount of Tris-HCl, 9 μl of 0.1 M CaCl 2 and 7.5 μl of 20% SDS were added and the volume was adjusted to 300 μl with H 2 O. Five units of nuclease Sa / 31 (New England, BioLabs) were added and the sample was ingested for 1 hour at 25 ° C. The reaction was terminated by extraction with 1 volume of phenol. The phenol phase was saved and re-extracted once with an equal volume of 10 mM Tris-HCl, pH 8.0, 1 mM EDTA. The protein-oligonucleotide complexes were then precipitated with the phenol phase by adding 10 volumes of acetone cooled on ice and placing on ice for 10 minutes. The pellet was dissolved in an SDS sample buffer and analyzed for SDS-PAGE. Gel drying and autoradiography were performed as described (Hsiang et al., J. Biol. Chem..260, 14873 (1985)). Example 11. Relaxation Analysis of Topoisomerase I The relaxation analysis was carried out as described by L.F. Liu and others, PNAS USA. 78, 3487 (1981). Briefly, each reaction mixture (20 μl) contained a mixture of relaxed and supercoiled YePg DNA (150 ng each) and 1 μl of Aspergillus topoisomerase I or human diluted to several degrees. After an incubation at 23 or 37 ° C for 15 minutes, the reactions were terminated by the addition of 5 μl of a pre-heated retention solution (5% sarcosyl, 25% sucrose, 50 mM EDTA and 0.05 mg / ml of bromophenol blue). The DNA samples were then analyzed using 1% agarose gel in TPE electrophoresis solution (90 mM Tris-phosphate, 2 mM EDTA, pH 8.0). Example 12. Topoisomerase I Cleavage Assay DNA topoisomerase I cleavage assays were carried out as described by Y. H. Hsiang et al., J. Biol. Chem., 260, 14873 (1985). The YEpG DNA was aligned with BamH1 and then labeled at the 3 'end with Klenow polymerase and [-32P] dCTP. After phenol extraction and ethanol precipitation, the labeled DNA was resuspended in 10 mM Tris, pH 8.0, and 1 mM EDTA. DNA separation analysis was performed in a reaction mixture (20 μl) containing 40 mM Tris-HCl, pH 7.8, 100 mM KCl, 10 mM MgCl 2, 0.5 mM dithiothreitol, 0.5 mM EDTA, 30 μg / ml of bovine serum albumin, 20 ng of labeled YEpG DNA and 1 ml of Aspergillus topoisomerase I or human diluted to several degrees. After incubation at 23 ° C for 15 minutes, the reactions were terminated by the addition of SDS (final concentration 1%) and proteinase K (final concentration 200 μg / ml). The proteinase K treatment was continued at 37 ° C for another hour. The finished reactions were denatured with alkali and then run (alkaline loading) were loaded directly into neutral charge buffer (neutral charge) on a 1% agarose gel in neutral TPE electrophoresis solution. Gel drying and autoradiography were carried out as described by Hsiang and others, cited above. Example 13. Yeast Cytotoxicity Analysis The topoisomerase one specific in vivo cytotoxicity assay was adapted from A. M. Knab et al., J. Biol. Chem., 268, 2232 (1993). In this system, several genes of topoisomerase I or cDNA were cloned into the single copy yeast plasmid vector (YCpGALI; Knab et al., Cited above) will be expressed under the control of GAL1 promoter in strain JN2-134 of S. cerevisiae (MAT, rd52 :: Leu2, trpl, ade2-1, his ?, ura3-52, sel, top1-1, and Ieu2; MA Bjornsti et al., Cancer Res .. 49. 6318 (1989)). The topoisomerase I gene or cDNA constructs in the vector are, respectively, the wild type yeast topoisomerase I gene (YCpGAL-ScTOPI; Kim &Wang, citation needed (1989)), a non-functional topoisomerase I gene in where tyrosine-727 was mutated in the active site to phenylalanine (YCpGAL1-Sctop1 Y727, AM Knab et al., J. Biol. Chem .. 268. 22322 (1993)) and wild-type human topoisomerase I DNA (YCp- GAL-hTOP1; MA Bjornsti et al., Cancer Res., 49, 6318 (1989)). To qualitatively test the topoisomerase I cytotoxicity and specificity of the drugs, yeast cells containing the specific plasmid were grown in drip medium supplemented with uracil, 2% galactose and the drug was tested. It has been established that the levator can survive when the topoisomerase I function is obliterated and that the topo I poisons only kill cells having a functional topoisomerase I. Therefore, the separation of the degree of relaxation of growth of each of the strains tested in the presence of several drugs with this in control plates (without drug) shows: (a) if the drug has such an effect of cytotoxities on yeast. , (b) if the cytotoxicity is specific for topoisomerase I; and (c) if there is a differential specificity of the drugs for yeast compared to topoisomerase I. Example 14. Characterization of Aspergillus nidulans Topoisomerase I The plasmid relaxant activity was used to monitor Aspergillus topoisomerase during purification. The relaxant activity in the Aspergillus cell extract was purified through a procedure designed for the purification of recombinant human DNA topoisomerase. coli (B. Gatto et al., Cancer Res., 56, 2795 (1996)). Several pieces of evidence suggest that the purified Aspergillus enzyme is the primary nuclear topoisomerase I identified and characterized in other eukaryotic organisms including yeast. First, the purified enzyme is highly active and represents the main DNA relaxant activity in the Aspergillus cell extract. Of two liters of culture, 30,000 units of topoisomerase I relax activity were obtained. Similar to human topoisomerase I, the Aspergillus enzyme relaxes the plasmid DNA to complete it and does not require Mg (I I) or a co-factor of energy. Second, the purified Aspergillus enzyme relaxes the supercoiled DNA side both negatively and positively, a property shared by all topoisomerase I of eukaryotic nuclear DNA. Third, the Aspergillus enzyme is sensitive to inhibition by camptothecin and H oechst 33342 (Ho33342) which are known to inhibit (poisonous) human nuclear topoisomerase I. The sensitivity of the enzyme Aspergillus to camptothecin and Hoechst 33342 was initially indicated by a phosphate transfer experiment that was designed to determine the approximate reduced molecular weight of the enzyme. In this experiment, 32 P-labeled DNA was reacted with Aspergillus topoisomerase I to form covalent protein DNA complexes. The covalent complex of A D N of topoisomerase I was directed with Ba / 31 to reduce the size of the labeled oligonucleotide which is covalently linked to topoisomerase I. Using this phosphate transfer method, Aspergillus topoisomerase I was identified as a 105 kDa protein that is slightly larger than recombinant human topoisomerase I (100 kDa). The lower band at the position of approximately 75 kDa is known to be a 100 kDa proteolytic degradation product of human topoisomerase I. The effect of the residual oligonucleotide on the mobility of topoisomerase I is apparently negligible. Recently, both camptothecin (100mM) and Ho33342 (1mM) simulated phosphate transfer as evidenced by the increasing labeling of Aspergillus topoisomerase I of 105 kDa. Higher concentrations of Ho33342, the phosphate transfer was progressively inhibited. This effect of camptothecin and Ho33342 is discussed below. Example 15. Camptothecin and Ho33342 Are Potent Inhibitors of Topoisomerase I from Asoeraillus. The phosphate transfer experiment suggested that both camptothecin and Ho33342 can inhibit Aspergillus topoisomerase I by a mechanism of poisoning. In order to test this possibility, Aspergillus topoisomerase I was used in a DNA separation reaction in the presence of several drugs. Both ptotecin (CPT) and Ho33342 (HOE) are potent inhibitors of Aspergillus topoisomerase I. Extensive DNA separation was observed at concentrations as low as 1.0 and 0.1 mg / ml for camptothecin and Ho33342, respectively. Increasingly, nitidine and coralline, which are known to be highly potent inhibitors of human DNA topoisomerase I, will not inhibit Aspergillus topoisomerase I to any significant degree. DM / l / 33, another highly potent inhibitor of human topoisomerase I, was only weakly inhibitory towards topoisomerase I of Aspergillus. Berenyl, which is inactive against human topoisomerase I was inactive against Aspergillus topoisomerase I. These results indicate that human and Aspergillus topoisomerase I are substantially different in terms of their sensitivity to several enzyme inhibitors. It is also interesting to note that at a higher concentration of Ho3342 (10 mg / ml), the DNA separation mediated by topoisomerase I was dramatically inhibited. This inhibitory effect of separation at higher concentrations of inhibitors has been previously described for a number of intercalators and minor slit DNA binding ligands (AY Chem et al., PNAS USA, 90. 8131 (1993) and was attributed to the inhibition of enzyme binding to the DNA standard (KM Tewey et al., Science, 226, 466 (1985).) The inhibitory effect of Ho33342 on phosphate transfer to Aspergillus topoisomerase I, therefore, can be explained similarly. Selective Sensitivity of Aspergillus topoisomerase I to Bi- and Ter-benzimidazoles Previous studies have identified a number of mono-, bi and ter-benzimidazoles as effective inhibitors (poisons) for mammalian DNA topoisomerase I. To test whether the topoisomerase Aspergillus I is also sensitive to the inhibitory effect of these benzimidazoles, a number of compounds were screened using separation analysis, Aspergillus topoisomerase I (60 units / reaction). tion) was strongly inhibited (poisoned) by 1_3 and 1_1, both of which are terbenzimidazoles. None of the monobenzimidazoles, including QS / ll / 50, QS / ll / 51, QS / II / 59A, a QS / ll / 9, exhibited any inhibitory effect on Aspergillus topoisomerase I. Previous studies have established that mono-benzimidazoles except QS / ll / 50, are inhibitors (poisons) of topoisomerase I of mammalian DNA. The selective sensitivity of topoisomerase I from Aspergillus to bi-benzimidazoles (e.g., Ho33342 and compound 2 where n is 3) and ter-benzimidazoles (e.g., 13 and 11), but not mono-benzimidazoles (v. .gr., QS / ll / 9) again indicates differences in drug sensitivity between human and Aspergillus enzymes. Example 17. Differences in Separation Specificity between human topoisomerase I and Asoeraillus. In addition to differences in drug sensitivity between topoisomerase I of humans and Aspergillus, additional differences in the specificity of separation have been observed between humans and topoisomerase I of Aspergillus. The enzyme patterns of humans (labeled hTOP1, 150 units / reaction) and Aspergillus (labeled AnTOPI, 60 units / reaction) are dramatically different in the presence of the benzimidazole Ho33342 (HOE). The largest number of separation sites and the greatest degree of separation exhibited by Aspergillus topoisomerase I in the presence of HOE are not understood. Although it is less obvious, the separation patterns of the enzymes of humans and Aspergillus were also different in the presence of camptothecin (CPT).

To regulate the possibility that contaminating topoisomerase II in the preparation of the enzyme Aspergillus toposiomerase I may contribute to the separation pattern, part of the samples were also analyzed for possible double-strand breaks, no double-strand DNA breaks were observed when the DNA samples were analyzed by neutral charges instead of alkalines. It is also evident from this experiment that Aspergillus topoisomerase I is less sensitive to C PT than the human enzyme. Differences in the specificity of separation between human enzymes (150 units / reaction) and Aspergillus (60 units / reaction) are also evident when terbenzimidazoles (13. and H) were used at 0. 1, 1.0 and 10. ug / ml. In addition, the Aspergillus enzyme appeared to be substantially less sensitive to 1_3 than the human enzyme. Example 18. Yeast Topoisomerase I Enzymes and Asperaillus Exhibit Similarity to Drug Susceptibility / Resistance Yeast top l suppression strains expressing human or yeast topoisomerase I under identical conditions have been used to assess drug sensitivity. differential of human and yeast enzymes (J. Nitiss et al., PNAS U SA 85, 7501 (1988); B, Gatto et al., Cancer Res. 56, 2795 (1996)). Although yeast cells expressing yeast topoisomerase I are sensitive to camptothecin, they are at least ten times more resistant to camptothecin than yeast cells expressing topoisomerase I from humans. Nitidine, DM / l / 33 and QS / l / 9 are highly cytotoxic against yeast cells expressing human topoisomerase I, but not cytotoxic for yeast cells expressing functional or non-functional yeast topoisomerase I. These results indicate that the topoisomerase I of yeast and Aspergillus are resistant to the same drugs (ie, nitidine, the protoberberin DM-l-33 and the mono-benzimidazole QS-l-9) that poison the topoisomerase I of humans. Therefore, Aspergillus topoisomerase I, similar to topoisomerase I of humans, is sensitive to the activity of camptothecin poisoning, the dibenzimidazoles H o33342 and terbenzimidazoles (ü and 13.). Although camptothecin appears to be less active against Aspergillus enzyme than human enzyme, terbenzimidazole seems to be more effective against Aspergillus enzyme than human enzyme. The effectiveness of terbenzimidazoles against Aspergillus topoisomerase I is not restricted to 1J_ and 1_3, the terbenzim idazoles of the formula I wherein n = 1, X = H, Ar = 5-phenyl, Y = H and Y 'is ethyl or -methoxyphenyl and 4-phenyl-isomer of compound 13, are also effective against the enzyme fu ngal in vitro. The overall superior sensitivity of topoisomerase I from Aspergillus to terbenzimidazoles is not understood. However, it is adjusted from the higher degree of separation and the looser sequence separation specificity, it can be argued that Aspergillus topoisomerase I may be less sensitive to the inhibitory effect of these DNA binding ligands. In other words, Aspergillus topoisomerase I can bind DNA with higher affinity than the human enzyme and is therefore less susceptible to the inhibitory effect of these binding ligands of A D N. The invention has been described with reference to several specific and preferred embodiments and techniques. However, it should be understood that many variations and modifications can be made while remaining within the spirit and scope of the invention.

Claims (50)

1. CLAIMS 1. A compound of the formula (I):
2. (I) wherein, Ar is an aryl of C6-C? 2, or heteroaryl of 5 to 12 members, comprising 1-3 N, S or O without peroxide, wherein each N is not substituted or substituted with H, alkyl of C? -C4 or benzyl; X is H, CN, CHO, OH, acetyl, CF 3, O-C 1 -C 4 alkyl, NO 2, NH 2, halogen or C 1 -C 4 haloalkyl; each Y is H, C1-C4 alkyl or aralkyl; Y 'is phenyl or methoxyphenyl; each Z is individually H, C? -C alkyl, halogen or C? -C4 haloalkyl; and n is 0 or 1; or a pharmaceutically acceptable salt thereof. 2. Claim 1, wherein Y 'is methoxyphenyl.
3. 3. Claim 1, wherein n is 1.
4. 4. Claim 1, wherein X is CN, CHO, OH, acetyl, CF3, O-C?-C, NO 2, NH 2, halogen or haloalkyl of C?-C4 and n is 0.
5. 5. Claim 1, wherein at least one Z is halogen or haloalkyl of C? -C4 and n is 0.
6. 6. Claim 1, 2, 3, 4 or 5, wherein medical therapy is the treatment of fungal infection.
7. 7. Claim 1, 2, 3, 4 or 5, wherein medical therapy is the treatment of cancer.
8. 8. The use of a compound of the formula (I): (I) wherein, Ar is an aryl of C6-C? 2, or heteroaryl of 5 to 12 members, comprising 1-3 N, S or O without peroxide, wherein each N is not substituted or substituted with H, alkyl of C? -C4 or benzyl; X is H, CN, CHO, OH, acetyl, CF 3, O-C 1 -C 4 alkyl, NO 2, NH 2, halogen or C 1 -C 4 haloalkyl; each Y is H, C? -C alkyl or aralkyl; Y 'is phenyl or methoxyphenyl; each Z is individually H, C? -C4 alkyl, halogen or C? -C haloalkyl; and n is 0 or 1; or a pharmaceutically acceptable salt thereof; for the manufacture of a medicine to treat fungal infection.
9. 9. Claim 8, wherein Y 'is methoxyphenyl.
10. 10. Claim 8, wherein n is 1.
11. 11. Claim 1, wherein X is CN, CHO, OH, acetyl, CF3, O-C-C4 alkyl, NO2, NH2, halogen or haloalkyl of C ? -C and n is 0.
12. 12. Claim 8, wherein at least one Z is halogen or haloalkyl of CrC and n is 0.
13. 13. The use of the compound of the formula (I): (i) wherein, Ar is an aryl of C6-C? 2, or heteroaryl of 5 to 12 members, comprising 1-3 N, S or O without peroxide, wherein each N is not substituted or substituted with H, alkyl of C? -C4 or benzyl; X is H, CN, CHO, OH, acetyl, CF3, O-C1-C4 alkyl, NO2, NH2, halogen or haloalkyl of C? -C4; each Y is H, C? -C alkyl or aralkyl; Y 'is phenyl or methoxyphenyl; each Z is individually H, C? -C4 alkyl, halogen or C? -C4 haloalkyl; and n is 0 or 1; or a pharmaceutically acceptable salt thereof; for the manufacture of a medicine for the treatment of cancer.
14. 14. Claim 13, wherein Y 'is methoxyphenyl.
15. 15. Claim 13, wherein n is 1.
16. 16. Claim 13, wherein X is CN, CHO, OH, acetyl, CF3, O-C?-C, NO 2, NH 2, halogen, or C-haloalkyl. C4 and n is 0.
17. 17. Claim 13, wherein at least one Z is halogen or C1-C4 haloalkyl and n is 0.
18. 18. A compound of Formula (I): (i) wherein, Ar is an aryl of C6-C? 2, or heteroaryl of 5 to 12 members, comprising 1-3 N, S or O without peroxide, wherein each N is not substituted or substituted with H, alkyl of C? -C or benzyl; X is H, CN, CHO, OH, acetyl, CF3, O-alkyl of d-C4, NO2, NH2, halogen or haloalkyl of C? -C4; each Y is H, C1-C4 alkyl or aralkyl; Y 'is phenyl or methoxyphenyl; each Z is individually H, C? -C4 alkyl, halogen or C? -C haloalkyl; and n is 0 or 1; or a pharmaceutically acceptable salt thereof.
19. 19. A compound of the formula (I): (I) wherein, Ar is a C6-C12 aryl, or 5- to 12-membered heteroaryl, comprising 1-3 N, S or O without peroxide, wherein each N is not substituted or substituted with H, C-alkyl? -C or benzyl; X is H, CN, CHO, OH, acetyl, CF 3, O-C 1 -C 4 alkyl, NO 2, NH 2, halogen or C 1 -C 4 haloalkyl; each Y is H, C? -C alkyl or aralkyl; Y 'is phenyl or methoxyphenyl; each Z is individually H, C? -C alkyl, halogen or C? -C4 haloalkyl; and n is 0 or 1; or a pharmaceutically acceptable salt thereof.
20. 20. A compound of the formula (I): (I) wherein, Ar is an aryl of C6-C? 2, or heteroaryl of 5 to 12 members, comprising 1-3 N, S or O without peroxide, wherein each N is not substituted or substituted with H, alkyl of C? -C4 or benzyl; X is H, CN, CHO, OH, acetyl, CF 3, O-C 1 -C 4 alkyl, NO 2, NH 2, halogen or C 1 -C 4 haloalkyl; each Y is H, C? -C4 alkyl or aralkyl; Y 'is phenyl or methoxyphenyl; each Z is individually H, C? -C4 alkyl, halogen or C? -C4 haloalkyl; and n is 0 or 1; or a pharmaceutically acceptable salt thereof.
21. 21. A compound of the formula (I): (I) wherein, Ar is an aryl of C6-C? 2, or heteroaryl of 5 to 12 members, comprising 1-3 N, S or O without peroxide, wherein each N is not substituted or substituted with H, alkyl of C? -C or benzyl; X is H, CN, CHO, OH, acetyl, CF 3, O-C 1 -C 4 alkyl, NO 2, NH 2, halogen or C 1 -C 4 haloalkyl; each Y is H, C? -C alkyl or aralkyl; Y 'is phenyl or methoxyphenyl; each Z is individually H, C? -C alkyl, halogen or C? -C4 haloalkyl; and n is 0 or 1; or a pharmaceutically acceptable salt thereof.
22. 22. A compound of the formula (I): (I) wherein, X is H, CN, CHO, OH, acetyl, CF3, O-C?-C4 alkyl, NO 2, NH 2, halogen or C?-C haloalkyl; each Y is H, C? -C4 alkyl or aralkyl; Y 'is phenyl or methoxyphenyl; each Z is individually H, C? -C4 alkyl, halogen or C? -C4 haloalkyl; and n is 0 or 1; or a pharmaceutically acceptable salt thereof.
23. 23. A compound of the formula (I): (I) wherein, X is H, CN, CHO, OH, acetyl, CF 3, O-C 1 -C 4 alkyl, NO 2, NH 2, halogen or C 1 -C haloalkyl; each Y is H, C? -C4 alkyl or aralkyl; Y 'is phenyl or methoxyphenyl; each Z is individually H, C? -C4 alkyl, halogen or C? -C4 haloalkyl, provides at least one Z is halogen or C1-C4 alkyl halo; and n is 0; or a pharmaceutically acceptable salt thereof.
24. 24. The compound of claim 18, 19, or 20 wherein n is 1.
25. 25. The compound of claim 21 or 22 wherein Ar is in the 5-position.
26. 26. The compound of claim 21 or 24, in where Ar is phenyl.
27. 27. The compound of claim 21 or 24 wherein Ar is 2-pyridyl.
28. 28. The compound of claim 18, 19, 20, 21, 22, 23 or 23, where X is halogen.
29. 29. The compound of claim 28, wherein X is Cl.
30. 30. The compound of claim 26, wherein X-Ar is p-chlorophenyl.
31. 31. The compound of claim 30, wherein each Y is H, and each Z is H.
32. 32. The compound of claim 18, 19, or 20, wherein n is 0.
33. 33. The compound of claim 32, wherein X is Cl.
34. 34. The compound of claim 33, where X is Br.
35. 35. The compound of claim 33 or 34, wherein Y 'is 4-methoxyphenyl; each Y is H, and each Z is H.
36. 36. The compound of claim 18, 19, 20, 21 or 22, wherein at least one Z is halogen or haloalkyl of C! -C4.
37. 37. The compound of claim 36, wherein at least one Z is F or CF3.
38. 38. The compound of claim 21 or 24, wherein Ar is benzo.
39. 39. The compound of claim 38, wherein Ar is 4,5-benzo.
40. 40. The compound of claim 38, wherein Ar is 5,6-benzo.
41. 41. A pharmaceutical composition comprising a compound of claim 18, 19, 20, 21, 22, 23 or 24 and a pharmaceutically acceptable carrier.
42. 42. A therapeutic method comprising treating fungal infection by administering to a mammal in need of such therapy, an effective amount of a compound of the formula (I): (I) wherein, Ar is an aryl of C6-C? 2, or heteroaryl of 5 to 12 members, comprising 1-3 N, S or O without peroxide, wherein each N is not substituted or substituted with H, alkyl of C? -C4 or benzyl; X is H, CN, CHO, OH, acetyl, CF3, O-C1-C4 alkyl, NO2, NH2, halogen or haloalkyl of C? -C4; each Y is H, C1-C4 alkyl or aralkyl; Y 'is phenyl or methoxyphenyl; each Z is individually H, C? -C4 alkyl, halogen or C? -C haloalkyl; and n is 0 or 1; or a pharmaceutically acceptable salt thereof.
43. 43. A therapeutic method comprising treating a fungal infection by administering to a mammal in need of such therapy a compound of claim 18, 19, 20, 21, 22, 23, or 24.
44. 44. The method of claim 42, in where the mammal is a human being.
45. 45. The method of claim 42, wherein the fungal infection is a fungal infection.
46. 46. The method of claim 42, wherein the compound is administered in combination with a pharmaceutically active carrier. 47. A therapeutic method comprising treating cancer by administering to a mammal in need of such therapy, an effective amount of a compound of the formula (I):
47. (I) wherein, Ar is an aryl of C6-C? 2, or heteroaryl of 5 to 12 members, comprising 1-3 N, S or O without peroxide, wherein each N is not substituted or substituted with H, alkyl of C? -C or benzyl; X is H, CN, CHO, OH, acetyl, CF 3, O-C 1 -C 4 alkyl, NO 2, NH 2, halogen or C 1 -C 4 haloalkyl; each Y is H, C ^ C- alkyl, or aralkyl; Y 'is phenyl or methoxyphenyl; each Z is individually H, C? -C4 alkyl, halogen or C? -C4 haloalkyl; and n is 0 or 1; or a pharmaceutically acceptable salt thereof.
48. 48. A therapeutic method comprising treating cancer by administering to a mammal in need of such therapy, an effective amount of a compound of claim 18, 19, 20, 21, 22, 23 or 24.
49. 49. The method of claim 47, in where the mammal is a human being.
50. 50. The method of claim 47, wherein the compound is administered in combination with a pharmaceutically acceptable carrier.