WO1999004789A1 - Substituted aurone derivatives - Google Patents

Abstract

A method for treating a microbial infection is disclosed. The method includes administering to a patient a pharmaceutical composition containing a compound of formula (IA) where each R is independently H, OH, Br, Cl, I, amino, thiol, nitro, C1-4 alkoxy, C1-4 alkenyloxy, C2-6 alkoxyalkyleneoxy, C1-4 alkylthio, C¿3-18 alkyl, or C¿3-18? alkenyl; or two adjacent R's, taken together, are a C2-18 bivalent moiety containing at least one oxygen atom, substituted or disubstituted with A or B or both, A being H, OH, Br, Cl, I, amino, or thiol, and B being H, C1-10 alkyl, C2-18 alkenyl, or C6-18 aryl; provided at least two Rs are not H; further provided that when each of two Rs is one of OH, C1-4 alkoxy, C1-4 alkenyloxy, or C2-6 alkoxyalkyleneoxy, and X is phenyl substituted with two substituents independently selected from OH, alkoxy, and alkenyloxy, the remaining R cannot be prenyl; further provided that when each of two Rs is one of OH, C1-4 alkoxy, C1-4 alkenyloxy, or C2-6 alkoxyalkyleneoxy, and X is phenyl substituted with three substituents independently selected from OH, alkoxy, and alkenyloxy, the remaining R cannot be prenyl; further provided that when each of two Rs is one of OH, C1-4 alkoxy, C1-4 alkenyloxy, or C2-6 alkoxyalkyleneoxy, and X is phenyl subsituted with a prenyl substituent and with two additional substituents independently selected from OH, alkoxy, and alkenyloxy, the remaining R cannot be H or OH; further provided that when each of two Rs is one of OH, C1-4 alkoxy, C1-4 alkenyloxy, or C2-6 alkoxyalkyleneoxy, and X is phenyl subsituted with a substituent containing three rings and with two additional substituents independently selected from OH, alkoxy, and alkenyloxy, the remaining R cannot be prenyl; X is C4-10 alkyl, C4-20 alkenyl, or a C4-20 single, C6-20 bridged, or C6-20 fused ring moiety containing cycloalkyl, cycloalkenyl, aryl, heterocycle, or heteroaryl, where X is substituted with H, OH, Cl, Br, I, amino, cyano, nitro, alkyl, alkoxy, alkenyl, or alkenyloxy; provided that if X is a heteroaryl or heterocyclic moiety where two Rs are each OH and meta to each other, then the remaining R is H and ortho to each of the two hydroxyls, and Y and Z are each O, and a ring atom of X is linked directly to the sp?2¿ carbon atom adjacent to X, then substituted with H, OH, Cl, Br, I, amino, cyano, alkyl, alkoxy, alkenyl, or alkenyloxy; and each of Y and Z is independently selected from O, S, and NH; or a pharmaceutically acceptable salt or ester thereof.

Description

SUBSTITUTED AURONE DERIVATIVES

Background of the Invention

The invention relates to methods of inhibiting microbial infections with substituted aurone derivatives.

Microbial infections, such as fungal infections and bacterial infections, can contribute to and complicate many diseases, including meningitis, pulmonary diseases, and respiratory tract diseases. Opportunistic infections have proliferated, particularly in immunocompromised patients, such as those with AIDS, those undergoing chemotherapy for cancer, and those undergoing therapy to prevent graft rejection following organ transplant surgery.

Fungal infections (mycoses) may be cutaneous, subcutaneous, or systemic. Superficial mycoses include tinea capitis, tinea corporis, tinea pedis, onychomycosis, perionychomycosis, pityriasis versicolor, oral thrush, and other candidoses such as vaginal, respiratory tract, biliary, eosophageal, and urinary tract candidoses. Systemic mycoses include systemic and mucocutaneous candidosis, cryptococcosis, aspergillosis, mucormycosis, paracoccidioidomycosis, North American blastomycosis, histoplasmosis, coccidioidomycosis, and sporotrichosis.

Pathogenic organisms include dermatophytes (e.g., Microsporum canis and other M. spp.; and Trichophyton spp. such as T. rubrum, and T. mentagrophytes), yeasts (e.g., Candida albicans or C. tropicalis), Torulopsis glabrata, Epidermophyton floccosum, Malassezia furfur (Pityropsporon orbiculare, or P. ovale), Cryptococcus neoformans, Aspergillus fumigatus and other Aspergillus spp., Zygomycetes (e.g., Rhizopus, Mucor), Paracoccidioides brasiliensis, Blastomyces dermatitidis, Histoplasma capuslatum, Coccidioides immitis, and Sporothrix schenckii.

Summary of the Invention In one aspect, the invention features a method for treating a microbial infection. The method includes administering to a patient a pharmaceutical composition containing a compound selected from formula (IA):

(IA) wherein each R is independently H, OH, Br, Cl, I, amino, thiol, nitro, C_1-4 alkoxy,

C,.₄ alkenyloxy, C₂„₆ alkoxyalkyleneoxy, C_M alkylthio, C₃.₁₈ alkyl, or C₃.₁₈ alkenyl; or two adjacent Rs, taken together, are a C_2-ι₈ bivalent moiety containing at least one oxgen atom, substituted or disubstituted with A or B, or both, A being H, OH, Br, Cl, I, amino, or thiol, and B being H, C ₀ alkyl, C₂.,₈ alkenyl, or C₆._lg aryl; provided that at least two Rs are not H; further provided that when each of two Rs is one of OH, C_M alkoxy, C alkenyloxy, or C₂.₆ alkoxyalkyleneoxy, and X is phenyl substituted with two substituents independently selected from OH, alkoxy, and alkenyloxy, the remaining R cannot be prenyl; further provided that when each of two Rs is one of OH, C]_₄ alkoxy, C,.₄ alkenyloxy, or C_2.6 alkoxyalkyleneoxy, and X is phenyl substituted with three substituents independently selected from OH, alkoxy, and alkenyloxy, the remaining R cannot be prenyl; further provided that when each of two Rs is one of OH, C,_₄ alkoxy, C,.₄ alkenyloxy, or C₂.₆ alkoxyalkyleneoxy, and X is phenyl substituted with a prenyl substituent and with two additional substituents independently selected from OH, alkoxy, and alkenyloxy, the remaining R cannot be H or OH; further provided that when each of two Rs is one of OH, C,.₄ alkoxy, C,_.4 alkenyloxy, or C₂.₆ alkoxyalkyleneoxy, and X is phenyl substituted with a substituent containing three rings and with two additional substituents independently selected from OH, alkoxy, and alkenyloxy, the remaining R cannot be prenyl; X is C₄.₁₀ alkyl, C₄.₂₀ alkenyl, or a C₄.₂₀ single, C₆.₂₀ bridged, or C₆.₂₀ fused ring moiety containing cycloalkyl, cycloalkenyl, aryl, heterocycle, or heteroaryl, wherein X is substituted with H, OH, Cl, Br, I, amino, cyano, nitro, alkyl, alkoxy, alkenyl, or alkenyloxy; provided that if X is a heteroaryl or heterocyclic moiety where two of R are each OH and meta to each other, the remaining R is H and ortho to each of the two hydroxyls, Y and Z are each O, and a ring atom of X is linked directly to the sp² carbon atom adjacent to X, then substituted with H, OH, Cl, Br, I, amino, cyano, alkyl, alkoxy, alkenyl. or alkenyloxy; and each of Y and Z is independently selected from O, S, and NH; or a pharmaceutically acceptable salt or ester thereof. The infection can be, for example, a fungal infection or a bacterial infection.

In another aspect, the invention features a compound selected from formulae (I)-(IV) below:

(HI) (IV) where

V is a bivalent C₂.₁₈ moiety containing at least one oxygen atom and substituted with A, B, or both; each of W and W is independently selected from the values for A, cyano, nitro, C,.₄ alkoxy, C,. alkenyloxy, C₂.₆ alkyloxyalkyleneoxy, C₂.₇ carboxyalkyloxy, C₇.₁₅ arylalkoxy, and C,.₄ alkylthio;

R_a is H, C_3-lg alkyl, C₃._lg alkenyl, C₅._lg cyclohexenyl, or C₆.₁₈ aryl; each of R_b and R_c is independently selected from H and C,.₄ alkyl;

X is substituted or unsubstituted C₃.₁₅ alkyl, C₃.₁₈ alkenyl, C₃._l5 cycloalkyl, C₄.,₅ cycloalkenyl, C₄.₂₀ bicyclo[a.b.c]alkyl, C₅.₂₀ bicyclo[a.b.c] alkenyl, C₈.₂₀ tricyclo[a.b.c.d]alkyl, C₈.₂₀ tricycloalkenyl, or C₂.₂₀ heterobicyclo[a.b.c]alkyl, or a combination thereof, where each of a, b, c, and d is independently 0 to 10 (e.g., 0 to 4, 0 to 6, or 1 to 7); and each of Y and Z is independently selected from O and S.

The invention also features synthetic methods suitable for combinatorial synthetic strategies for the production of diverse libraries of structurally related compounds. Other features and advantages of the invention will be apparent from the following detailed description, and from the claims.

Detailed Description In one aspect, the invention features a method of inhibiting a microbial infection, wherein the compound of formula (IA) is selected from formulae (I)-(IV):

<πi) (rv)

In one aspect, V is a bivalent C₂.₁₈ moiety containing at least one oxygen atom and substituted with A, B, or both. V can contain between 1 and 3 rings, e.g., 1 ring, 2 rings, or three rings. For example, V can be selected from the following five formulae:

Each of W and W is independently selected from the values for A, cyano, nitro, C alkoxy, C_M alkenyloxy, C_2.6 alkyloxyalkyleneoxy, C₂.₇ carboxyalkyloxy, C-₇.,₅ arylalkoxy, and C,.₄ alkylthio.

R_a is H, C₃._Ig alkyl, C_3.I8 alkenyl, C₅.,_g cyclohexenyl, or C₆._lg aryl. For example, R,, is H, prop-2-enyl, cinnamyl, 2-methylprop-2-enyl, but-2-enyl, 3-methylbut-2-enyl, 3,7-dimethylocta-2,6-dienyl, (cyclohexenyl)methyl, 3,7,1 l-trimethyldodeca-2,6.10-trienyl, or benzyl. In some cases, 1^ is not prenyl or isoprenyl.

Each of R_b and R,. is independently selected from H and C,.₄ alkyl. In one method, the compound can be of the formula Q=(CHX) where Q is derived from the benzofuranone analogs or derivatives from Schemes Q-l through Q-l 1, and the geometry of the double bond is E or Z. In Schemes Q-l through Q-l 1, the compounds are of the formula Q-H₂, where the two hydrogens are methylene hydrogens.

In another aspect, the compound has an IC₅₀ of less than 50 micrograms per milliliter against at least one pathogenic strain of Candida or Aspergillus. In one embodiment, the compound is of formula (III), where each of Y and

Z is independently selected from O and S, for example, formulae S01-S06 and S08-S19 of Scheme P-l. Other embodiments include a compound where: W and W are selected from H, OH, methoxy, methoxymethyleneoxy, and carboxyrαethoxy; where Y and Z are O, and at least one of W and W is OH; where X is a heterocyclic radical, e.g., a heteroaryl; where X is C₄.₁₀ alkyl, C₄.₂₀ alkenyl, or a C₄.₂₀ single, C₆.₂₀ bridged, or C₆.₂₀ fused ring moiety containing cycloalkyl, cycloalkenyl, or aryl; where X is a nonaromatic moiety containing cycloalkyl, cycloalkenyl, alkyl, or alkenyl; or where the compound is selected from S12 and S02.

Examples of X include benzyl, 2,5-dimethoxyphenyl, 2,3-dimethyl-4-methoxyphenyl, 3-benzyloxyphenyl, 3-phenoxyphenyl,

4-benzyloxy-3-methoxyphenyl, 4-[3-propenoic acid]-phenyl, 2-ethoxy-l-naphthyl,

1 -(methylthio)ethyl, DL-1-phenylethyl, 4-n-pentyloxyphenyl,

1 -(phenylsulfonyl)-2-pyrrolyl, 4-(3-dimethylaminopropoxy)phenyl, 3-phenylpropyl, 2,4-diethoxy-m-tolulyl, 2,6,6-trimethylcyclohexene-l -methyl,

2,5-dimethoxy-3-tetrahydrofuranyl, 4-methyl-5-imidazolyl, 4-n-pentylphenyl,

2-benzyloxy-4,5-dimethoxyphenyl, 1 -pyrenyl, 3,5-dibenzyloxy-3-methoxyphenyl,

3-methyl-4-methoxyphenyl, 4-n-decyloxyphenyl, 2,4-dimethoxy-3-methylphenyl, t-butyl, 3-(4-t-butylphenoxy)phenyl, 2-n-hexyloxyphenyl, 2-(4-chlorophenylthio)phenyl, cyclopropyl, 2,6-dimethoxy-4-hydroxyphenyl,

4-benzyloxyphenyl, 2-benzyloxyphenyl, 8-hydroxy -l,l,7,7-tetramethyljulolidin-9-yl,

2,3,6,7-tetrahydro-8-hydroxyjulolidin-9-yl,

2-methoxymethyl-l-pyrrolidinyl, 5-(2-nitrophenyl)furanyl,

1 , 1 -dimethyl-2-hydroxyethyl, 5-methylfuranyl, 5-(3-chlorophenyl)furanyl, 2,4-hexadienyl, 5-[3(trifluoromethyl)-phenylfuranyl], 4,5-dimethyl-4-pentenyl, imidazolyl, ferrocenyl, 2,6-dimethylhept-5-enyl,

5-[2-(trifluoromethyl)-phenyl]furanyl, 5-(hydroxy-2-nitromethyl)furanyl,

2,4-dimethyl-2,6-heptadienyl, 1-phenylethyl, 5-(2-chlorophenyl)furanyl, benzyl,

5-ethyl-2-furanyl, 5-(4-nitrophenyl)-furanyl, pentamethylphenyl, 1 -(methyldithio)isopropyl, 4-trifluoromethylphenyl, 3-fluoro-4-methoxyphenyl, or the

X of a compound of Schemes X-l through X-10, wherein the compounds of Schemes

X-l through X-10 have the formulae X-CHO.

The fungal infection can be: an infection of a Candida species, an infection of a fungus resistant to at least one azole antifungal agent (e.g., where the azole antifungal agent is fluconazole); or an infection of an Aspergillus species.

Examples of pathogen strains include C. albicans, C glabrata, C krusei, C tropicalis, C parapsilosis, A. fumigatus, and A. niger.

The invention also features aurone derivatives, such as those described in formulae (I)-(IV) in the Summary section. Examples of these compounds include those where X is C₃.₁₅ alkyl, C₃.₁₈ alkenyl, C₃.₁₅ cycloalkyl, C₄.₁₅ cycloalkenyl, C₅_₁₀ bicyclo[a.b.c]alkyl, C₅.₁₀ bicyclo[a.b.c]alkenyl, C₈.₂₀ tricyclo[a.b.c.d]alkyl, C₈_₂₀ tricycloalkenyl, C₃.₁₀ heterobicyclo[a.b.c]alkyl, or a combination thereof, where each of a, b, c, and d is independently 0 to 6; X is C₃.₁₅ alkyl, C₃._lg alkenyl, C₃.₁₅ cycloalkyl, or C₄.,₅ cycloalkenyl; where X is C₅.₁₀ bicyclo[a.b.c.]alkyl, C₅.₁₀ bicyclo[a.b.c]alkenyl, C₈.₁₅ tricyclo[a.b.c.d] alkyl, C_g._I5 tricycloalkenyl, C₃.₁₀ heterobicyclo[a.b.c]alkyl, or a combination thereof; where each of W and W is independently selected from H, hydroxyl, methoxy, hydroxymethyl, and halomethyl; and where W and W are both hydroxyl; or a combination thereof. The bridges can be ortho-fused or ortho- and peri-fused. The bridge can be alkylene, azo, azimino, biimino, epidioxy, nitrilo, imino, furano, epoxythioxy, epithio, alkanoxy, epoxy, or alkanoxyalkano (e.g., methanoxymethano). The invention also features additional novel compounds described in the above method of treatment.

In some embodiments, enantiomers of disclosed compounds are separated. The bridging olefmic bond between Q and X is sometimes preferably E (entgegen) and sometimes preferably Z (zusammen). Depending on the individual embodiment, chiral centers may be (R) or (S).

Terms

Some terms are defined below, and some terms are defined elsewhere in the disclosure.

Alkyls may be substituted or unsubstituted and may be straight, branched, or cyclic. Preferably, alkyl groups have between 1 and 10 carbon atoms, and more preferably have between 1 and 6 carbon atoms. Examples of alkyls include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, t-butyl, pentyl, cyclopentyl, isopentyl, neopentyl, t-pentyl, sec-pentyl, hexyl, cyclohexyl, isohexyl,

2,3-dimethylbutyl, 2,2-dimethylbutyl, 3-ethylpentyl, 3,4-dimethylpentyl, heptyl, octyl, nonyl, decyl, and (2,3,4-trimethylcyclo-hexyl)methyl. An alkylene is a bivalent hydrocarbon, e.g., an alkyl group with an additional hydrogen removed, such as methylene, propylene, or 1 ,4-cyclohexylene. Alkoxy groups are alkyl groups linked to the remainder of the molecule, e.g., a ring, by an oxygen. Alkoxy groups also include polyethers, such as methoxyethyloxy. Alkyl, alkenyl, alkynyl, aryl, and heterocyclic radicals, whether or not substituting groups, (discussed below) may be linked by alkyl, alkenyl, alkynyl, ether, ester, amide, urea, urethane, amino, thioether, or thioester groups, such as methoxymethyl and alkylthioalkyl.

Alkenyls are alkyl groups with one or more unsaturated carbon-carbon bonds, such as cyclopentenyl, cyclopentadienyl, cyclohexadiene, but-2-enyl, 3,4-dimethylpent-3-enyl, allyl, vinyl, prenyl, isoprenyl, and norbornenyl. Examples of alkenylenes include vinylene and propenylene. Similarly, alkynyl groups have one or more triple bonds, and may also include one or more double bonds.

Aryls include aromatic rings, substituted or unsubstituted, preferably having between 6 and 20 carbon atoms, and more preferably between 6 and 14 carbon atoms, exclusive of substitution on the ring. Examples of aryls include phenyl, naphthyl, indenyl, pentalenyl, anthryl, azulyl, and biphenylyl. Combinations include alkylaryls (e.g., tolyl, xylyl, mesityl, cumenyl, 2-ethyl-4 methylphenyl) and arylalkyls (e.g., benzyl, phenylethyl, ) or arylalkenyls, and divalent arylenes such as 1 ,4-phenylene.

Haloalkyl (or haloalkenyl or haloalkynyl) includes any alkyl (or alkenyl or alkynyl) group where at least one hydrogen is replaced with a halogen (fluorine, chlorine, bromine, or iodine). Where more than one hydrogen is replaced (e.g., a dihaloalkyl or a hexahaloalkyl), the halogens are selected independently and may be on the same carbon atom or on different carbon atoms. Amino-substituted, nitro-substituted, or otherwise substituted alkyls (or alkenyls or alkynyl or aryls) are analogous to the above. Halomethyls include perchloromethyl, bromomethyl, and fluorochloromethy 1.

Heterocyclic radicals may be aromatic (heteroaryl) or nonaromatic, and substituted or unsubstituted. They have one, two or three rings which are single, fused, bridged rings, or polycyclic. They contain between 2 and 15 carbon atoms in the ring, i.e., exclusive of substitution. They can be linked to the rest of the molecule through a carbon atom or a heteroatom. Heterocyclic radicals include thienyl, thianthrenyl, furanyl, pyranyl, isobenzofuranyl, chromenyl, xanthenyl, phenoxathiinyl, 2H-pyrrolyl, pyrrolyl, imidazolyl, pyrazolyl, isothiazolyl, isoxazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, indolizinyl, isoindolyl, quinuclidinyl, 3H-indolyl, indolyl, indazolyl, purinyl, 4H-quinolizinyl, isoquinolyl, quinolyl, phthalazinyl, naphthyridinyl, quinoxalinyl, cinnolinyl, pteridinyl, 4H-carbazolyl, carbazolyl, beta-carbolinyl, phenanthridinyl, acridinyl, perimidinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxazinyl, isochromanyl, chromanyl, furazanyl, pyrrolinyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrazolinyl, imidazolinyl, piperidyl, piperazinyl, and morpholinyl. Heterocyclic radicals also include benz[h]isoquinolinyl, thieno[2,3-b]furanyl, 2H-furo[3,2-b]pyranyl.

Substituted moieties have one, two, three, or more of the following independently selected substituting moieties (instead of a hydrogen): C,.₁₀ alkyl, C₂.₁₀ alkenyl, C _]0 alkoxy, C₂.₁₀ alkenyloxy, C,.₁₀ haloalkyl, Cj_₁₀ haloalkoxy, aryl, aryloxy, hydroxy, nitro, chloro, fluoro, bromo, and iodo, thiol, cyano, and amino. Substituting moieties also include combinations of the above with carbonyl (acyl), sulfonyl, thionyl (e.g., thioketone), and carboxyl, such as alkyloxycarbonyl, arylalkyloxy, (N,N-dialkylamino)alkoxy, arylsulfonyl, and carboxylic acids. In some embodiments, substituting moieties have between 1 and 6 carbon atoms, and more preferably have between 1 and 3 carbon atoms. Examples of carbon-containing substituting moieties include chloromethyl, hydroxymethyl, bromoethyl, methoxy, and ethoxy. An alkyl does not have an alkyl or haloalkyl substituent, although, for example, a cycloalkyl may have an alkyl or haloalkyl substituent. The invention also encompasses compounds identical to any of the disclosed structures (e.g., formula (IV)), except that one or more conventional protecting groups are used, such as hydroxyl protecting groups, carboxylate protecting groups, and carbonyl protecting groups. Methods of adding and removing such protecting groups are well known in the art (see, for example, Protective Groups in Organic Synthesis, by T.W. Greene and P.G.M. Wuts, 2nd ed., 1991, Chapters 2-5). For example, the following representative hydroxyl protecting groups are provided. There is some overlap between the above-described R moieties and the disclosed hydroxyl protecting groups. Methyl ethers include methoxymethyl; methylthiomethyl; t-butylthio- methyl; (phenyldimethyldiyl)methoxy-methyl; benzyloxymethyl; p-methoxybenzyl- oxymethyl; (4-methoxyphenoxy)methyl; guaiacolmethyl; t-butoxymethyl; 4- pentenyloxymethyl; siloxymethyl; 2-methoxyethoxym ethyl; 2,2,2-trichloro- ethoxymethyl; bis(2-chloroethoxy)methyl; 2-(trimethylsilyl)ethoxymethyl; tetrahydropyran-2-yl; 3-bromotetrahydropyran-2-yl; 1 -methoxycyclohexyl; 4- methoxy-tetrahydropyran-2-yl; 4-methoxytetrahydrothiopyran-2-yl; 4- methoxytetrahydrothio-pyran-2-yl-S,S-dioxido; l-[(2-chloro-4-methyl)phenyl]-4- methoxypiperidin-4-yl; l,4-dioxan-2-yl; tetrahydrofuranyl; tetrahydrothiofuranyl; and 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl. Ethyl ethers include 1-ethoxy ethyl; l-(2-chloroethoxy)ethyl; 1 -methyl- 1- methoxyethyl; 1 -methyl- l-benzyloxy-2-fluoroethyl; 2,2,2-trichloroethyl; 2-trimethylsilylethyl; 2-(phenylselenyl)ethyl; t-butyl; allyl; p-chlorophenyl; p- methoxyphenyl; and 2,4-dinitrophenyl.

Benzyl ethers include benzyl; p-methoxybenzyl; 3,4-dimethoxybenzyl; o- nitrobenzyl; p-nitrobenzyl; p-halobenzyl; 2,6-dichlorobenzyl; p-cyanobenzyl; p-phenylbenzyl; 2- and 4-picolyl; 3-methyl-2-picolyl-N-oxido; diphenylmethyl; p,p'- dinitrobenzhydryl; 5-dibenzosuberyl; triphenylmethyl; α-naphthyldiphenylmethyl; p- methoxyphenyldiphenylmethyl; di(p-methoxyphenyl)phenylmethyl; tri(p- methoxyphenyl)methyl; 4-(4'-bromo-phenacyloxy)phenyldiphenylmethyl; 4,4',4"- tris(4,5-dichlorophthalimidophenyl)methyl; 4,4',4"-tris-(levulinoyloxyphenyl)methyl; 4,4',4"-tris(benzoyloxyphenyl)methyl; 3-(imidazol-l-ylmethyl)bis(4',4"- dimethoxyphenyl)-methyl; l,l-bis(4-methoxyphenyl)-l'-pyrenylmethyl; 9-anthryl; 9-(9-phenyl)xanthenyl; 9-(9-phenyl-10-oxo)anthryl; l,3-benzodithiolan-2- yl; and benzisothiazolyl S,S-dioxido. Silyl ethers include trimethylsilyl; triethylsilyl; triisopropylsilyl; dimethylisopropylsilyl; diethylisopropyl-silyl; dimethylthexylsilyl; t- butyldimethylsilyl; t-butyl-diphenylsilyl; tribenzylsilyl; tri-p-xylylsilyl; triphenyl- silyl; diphenylmethylsilyl; and t-butylmethoxyphenylsilyl. Esters include formate; benzoylformate; acetate; chloroacetate; trichloroacetate; methoxyacetate; triphenylmethoxyacetate; phenoxyacetate; p- chlorophenoxyacetate; p-(phosphate)phenylacetate; 3-phenylproprionate; 4- oxopentanoate (levulinate); 4,4-(ethylenedithio)pentanoate; pivaloate; adamantoate; crotonate; 4-methoxycrotonate; benzoate; p-phenylbenzoate; and 2,4,6- trimethylbenzoate.

Carbonates include methyl carbonate; 9-fluorenyl-methylcarbonate; ethyl carbonate; 2,2,2-trichloroethyl carbonate; 2-(trimethylsilyl)ethyl carbonate; 2-(phenyl- sulfonyl)ethyl carbonate; 2-(triphenylphosphono)ethyl carbonate; isobutyl carbonate; vinyl carbonate; allyl carbonate; p-nitrophenyl carbonate; benzyl carbonate; p- methoxybenzyl carbonate; 3,4-dimethoxybenzyl carbonate; o-nitrobenzyl carbonate; p-nitrobenzyl carbonate; S-benzyl thiocarbonate; 4-ethoxy-l -naphthyl carbonate; and methyl dithiocarbonate.

Protecting groups with assisted cleavage include 2-iodobenzoate; 4-azidobutyrate; 4-nitro-4-methylpentanoate; o-(dibromomethyl)benzoate; 2-formylbenzenesulfonate; 2-(methylthiomethoxy)ethyl carbonate;

4-(methylthiomethoxy)-butyrate; and 2-(methylthiomethoxymethyl) benzoate.

Miscellaneous esters include 2,6-dichloro-4-methylphenoxyacetate; 2,6- dichloro-4-( 1,1,3 ,3-tetramethyl-butyl)phenoxyacetate; 2,4-bis( 1 , 1 -dimethylpropyl)- phenoxy-acetate; chlorodiphenylacetate; isobutyrate; monosuccinoate; (E)-2-methyl- 2-butenoate (tigloate); o-(methoxycarbonyl)benzoate; p-benzoate; α-naphthoate; nitrate; alkyl N,N,N',N'-tetramethylphosphorodiamidate; N-phenylcarbamate; borate; dimethylphosphinothioyl; and 2,4-dinitrophenyl-sulfenate.

Sulfonates include methanesulfonate (mesylate); benzylsulfonate; and tosylate. Cyclic acetals and ketals include methylene; ethylidene; 1-t- butylethylidene; 1-phenylethylidene; 4-methoxyphenylethylidene; 2,2,2- trichloroethylidene; acetonide (isopropylidene); cyclopentylidene; cyclohexylidene; cycloheptylidene; benzylidene; p-methoxybenzylidene; 2,4-dimethoxybenzylidene; 3,4-dimethoxybenzylidene; and 2-, 3-, or 4-nitrobenzylidene.

Cyclic ortho esters include methoxymethylene; ethoxymethylene; dimethoxymethylene; 1-methoxyethylidene; 1 -ethoxyethylidine; 1 ,2-dimethoxy- ethylidene; α-methoxybenzylidene; l-(N,N-dimethylamino)ethylidene derivative; α-(N,N-dimethylamino)benzylidene derivative; and 2-oxacyclo-pentylidene.

Note that these cyclic ortho esters, like the bivalent organic moieties recited above for adjacent pairs of substituents (e.g., R, and R₂ in formula (IV)), may react with non-adjacent hydroxyl moieties. For example, a bivalent organic moiety recited in the preceding paragraph or recited above for adjacent pairs of substituents may be selected for two nonadjacent substituents on the same molecule or for any two substituents on two separate molecules. The two separate molecules can be the same or different, and are selected from compounds disclosed herein.

Silyl derivatives include di-t-butylsilylene group; 1, 3-(l, 1,3,3- tetraisopropyldisiloxanylidene) derivative; tetra-t-butoxydisiloxane- 1 ,3-diylidene derivative; cyclic carbonates; cyclic boronates; ethyl boronate; and phenyl boronate. Preferred protecting groups for catechols include cyclic acetals and ketals such as methylene, acetonide, cyclohexylidene, and diphenylmethylene; and cyclic esters such as cyclic borate and cyclic carbonate.

The invention encompasses other C,_₁₀ hydroxyl protecting groups not individually identified above which are pharmaceutically acceptable, and are optionally metabolized (e.g., cleaved or modified) to form one of the compounds disclosed herein. In other words, the invention encompasses metabolic precursors of the disclosed compounds and metabolites of the disclosed compounds having antimicrobial activity. The invention also encompasses amides, amine salts, and other organic salts of the disclosed compounds. Amides may be formed by reacting a disclosed compound or activated derivative thereof with any naturally-occurring amino acid, an oligopeptide having up to 10 (e.g., 4, 3, or 2) residues, a peptidomimetic having a molecular weight less than 300, or any C,.₂₀ organic moiety having an amino group that is not already described above. The term "naturally occurring amino acid" is meant to include the 20 common α-amino acids (Gly, Ala, Val, Leu, He, Ser, Thr, Asp, Asn, Lys, Glu, Gin, Arg, His, Phe, Cys, Trp, Tyr, Met and Pro), and other amino acids that are natural products, such as norleucine, ethylglycine, ornithine, methylbutenylmethylthreonine, and phenylglycine. Examples of amino acid side chains include H (glycine), methyl (alanine), -CH₂-(C=O)-NH₂ (asparagine), -CH₂-SH (cysteine), and -CH(OH)CH₃ (threonine).

Subjects or patients of the disclosed methods may be any living animal, plant, or plant product (e.g., grain or feed). Animals include mammals, particularly humans. Animals also include domestic animals bred for food or as pets, such as horses, cows, sheep, poultry, fish, pigs, cats, dogs, and zoo animals. Plants include trees, crops, grasses, and flowering plants.

r.

<o

CΛ o o rO

n

Scheme Q- / J oo oo oo

PSS057 o as as

O

Scheme X-l

Scheme X-3

Scheme X-4

Scheine X-b

Scheme X-

C

Scheme X-S

Sche e X-9

Scheme X-10

Matrix Synthesis

Over 3000 aurones were synthesized by matrix methodology and screened for antimicrobial activity. Numerous benzofuranones (Schemes Q-l through Q-l 1) and aldehydes (Schemes X-l through X-10) were obtained commercially or synthesized. One-hundred and sixty aldehydes were purchased.

The aldehydes were divided into two sets or plates of 80 aldehydes. Each set was reacted with a given benzofuranone. Phenolic hydroxyl groups were protected, for example, as methoxymethyl ethers. In general, the alicyclic or aliphatic aldehydes were less reactive than the aromatic aldehydes and thus required more vigorous conditions, such as higher temperatures. Even so, yields were generally lower than the aromatic aldehydes. Portions of the adducts were screened for antimicrobial activity, and the remainder deprotected with trimethylsilyl chloride in methanol to yield the unprotected aurone, which was also screened for antimicrobial activity. Specifically, a solution of benzofuranone in methanol, (1 M, 10 μl) was added to a 2 ml polypropylene tube containing 100 μl of methanol. After a solution of methanolic sodium methoxide (0.5 M, 22 μl) was added, the reaction was shaken for 1 minute. A solution of the aldehyde in methanol (1 M, 10 μl) was added and the reaction was left shaking for 5 minutes. After partitioning between ethyl acetate (1 ml) and water (0.5 ml), the organic layer was collected and transferred to another 2 ml polypropylene tube and allowed to dry. The dried sample was redissolved in 1 ml methanol and divided into 2 equal portions, one of which was dried and tested for antimicrobial activity. To the second portion was added trimethylsilyl chloride (50 μl). After standing at room temperature for 8 hours, ethyl acetate (1 ml) and saturated sodium bicarbonate (500 μl) were added. The organic layer was collected and dried. The dried sample was tested for antimicrobial activity. The compounds were tested for antimicrobial activity using, for example, methods described in WO 97/26873. Antifungal Activity

Thirty-eight 96-well plates were prepared with the above samples and tested against both C. albicans and A. fumigatus at either 8 μg/ml or 12.5 μg/ml. Plates with high activity were retested at the same concentration to confirm activity. Based on the above, preferred compounds have at least one phenolic hydroxyl group on the benzofuranone portion of the aurone, preferably at position 5 (W in formulae I-IV). Compounds with 2,3-dihydroxyphenyl or 2,3,4-trihydroxyphenyl (derived from the aldehyde reagent) exhibited good inhibition against Candida, yet exhibited little inhibition of Aspergillus . Some preferred anti-Candida compounds have polar substituents on the aldehyde portion of the aurone. Turning to an -Aspergillus compounds, aurones including 3,5-di-t-butyl-4-hydroxyphenyl and 2,4-difluorophenyl (again derived from the aldehyde reagent) generally exhibited good inhibition. Less polar substituents tend to improve inhibition of Aspergillus. Inhibition can be measured in terms of an IC₅₀, an MIC, or a percent inhibition relative to control (absence of test compound) at a given concentration, such as 8 μg/ml or 12.5 μg/ml. In general, a percent inhibition of at least 30% at 8 μg/ml or 12.5 μg/ml is preferred (e.g., at least 40%, at least 50%, at least 65%, and least 70%, and at least 85%). Preferred compounds of formula (II) exhibit a percent inhibition of at least 70%, including two as high as 87%. These compounds have formulae where W is OH, V is 3',4'-(l,l-dimethylpropyloxy) to form a dihydropyran series, and X is selected from 3,4-dimethoxyphenyl, 4-t-butylphenyl, 2(prop-2-enyloxy)phenyl, 3-phenoxyphenyl, 3-ethoxy-4-hydroxyphenyl, 6-cyclohexenyl, and norborn-4-yl. The inhibition values of 60 compounds are shown in Tables 1 through 3 on the next three pages.

CA 49% CA 92% CA 0% CA 94% CA 89%

AF 40% AF 0% AF 0% AF 31% AF 0%

@ 12.5μg/ml ¹ 12.5μg/ml 12.5μg/ml ¹ 12.5μg/ml ^■ 12.5μg/ml

CA 0% CA 95% CA 57% CA 79% CA 94%

AF 43% AF 36% AF 48% AF 27% AF 44%

^@ 12.5μg/ml ' 12.5μg/ml 12.5μg/ml 12.5μg/ml 12.5μg/ml

CA 67% CA 87% CA 80% CA 96% CA 0%

AF 94% AF 0% AF 76% AF 52% AF 69%

@ 8μg/ml @ 8μg/ml @ 8μg/ml @ 8μg/ml @ 8μg/ml

CA 53% CA 0% CA 52% CA 45% CA 69%

AF 96% AF 0% AF 68% AF 0% AF 35%

@ 8μg/ml D 8μg/ml @ 8μg/ml @ 8μg/ml @ 8μg/ml

Antibacterial Activity

Compounds described herein also have activity against bacteria. The compounds shown in the table below were tested using assays known in the art and were found to have activity against Staphylococcus aureus.

Use

Compositions including one or more disclosed compounds are useful for inhibiting microbial infections, or combinations of infections. The invention features a method for inhibiting a microbial infection in a subject, which method includes administering a pharmaceutically effective amount of such a composition. One aspect is a method for inhibiting a fungal infection which is resistant or sensitive to known therapies, such as fluconazole or other azoles. Examples of fluconazole-resistant or fluconazole-sensitive strains include C. glabrata, C kefyr, and C. tropicalis.

Formulation and Administration A disclosed composition contains from about 0.1 to 90% by weight (such as about 0.1 to 20%, or about 0.5 to 10%) of active compound(s). Disclosed compositions can be formulated as solids or liquids for oral administration, or as liquids or semi-solids (ointments, creams) for topical administration. The compositions can also be formulated for administration by nebulization or inhalation, or administration by intravenous, intramuscular, or intraperitoneal injection. Formulations for controlled release, including implantable or biodegradable or biocompatible matrices, are also contemplated. Controlled release includes continuous and intermittent release. Methods of formulation, including pharmaceutical carriers, are well-known to those in the art. The effective amount of active compound(s) used to practice the present invention for therapeutic or prophylactic treatment of conditions caused by or contributed to by a microbial infection varies depending upon the manner of administration, the age, body weight, and general health of the subject. Ultimately, the attending physician or veterinarian will decide the appropriate amount and dosage regimen. Such amount is referred to as an "effective" amount.

There now follow particular examples that describe the preparation of compounds of the invention, and the methods of the invention. These examples are provided for the purpose of illustrating the invention, and should not be construed as limiting.

Example 1 : 2-Amino-4.6-dimethoxy-κ-chloroacetophenone (5)

Boron trichloride (1M in CH₂C1₂, 6.5 ml) was added to a dry N₂ flushed flask through a septum and cooled on ice. 3,5-Dimethoxyaniline (1.0 g, 6.5 mmol), dissolved in dry CH₂C1₂, was added, followed by dropwise addition of chloroacetonitrile (0.500 ml, 7.8 mmol). The mixture was stirred under N₂ for 10 minutes, and ZnCl₂ (0.98 g, 7.2 mmol) was added. The green mixture was refluxed for 1 hour and stirred 18 hours at room temperature. Hydrochloric acid (2N, 5 ml) was added; the mixture was refluxed for 30 minutes, allowed to cool to 25 °C, and an excess of NaOH (2N) was added. After two extractions with CH₂C1₂, the organic phases were dried (MgSO₄), filtered, and concentrated in vacuo to afford the crude product (1.013 g, 68%). Purification by column chromatography (eluent: CH₂Cl₂/MeOH: 99/l) gave (5) (834 mg, 56%). HPLC-MS: MY230). 'H-NMR (CDC1₃, 250 MHz): δ 6.5 (broad s, 2 H); 5.65 (m, 2 H); 4.70 (s, 2 H); 3.79 (s, 3 H); 3.72 (s, 3 H).

Example 2: 4.6-Dimethoxy-3-indolinone (6)

2-Amino-4,6-dimethoxy- -chloroacetophenone (670 mg, 2.9 mmol) was dissolved in dry acetone (10 ml). K₂CO₃ (604 mg, 4.4 mmol) and a little KI was added, and the mixture was refluxed for 4 hours, then stirred at room temperature for 2 days until the starting material had disappeared. The solvent was evaporated in vacuo, and water (20 ml) was added to the compound. Extraction with CH₂C1₂, drying with MgSO₄, filtration, and evaporation of the solvent in vacuo afforded the crude product. Purification by column chromatography (eluent: CH₂Cl₂/MeOH: 99/1) gave (6) (394 mg, 70%) as green crystals. 'H-NMR (CDC1₃, 250 MHz): δ 6.5 (broad s, 2 H); 5.65 (dd, 2 H); 4.39 (s, 2 H); 4.39 (s, 2 H); 3.82 (s, 3 H); 3.72 (s, 3 H). Example 3: 2-Amino-4.5.6-trimethoxy- -chloroacetophenone 8

The target compound (8) was synthesized in the manner described for the synthesis of (5) using 3,4,5-trimethoxyaniline (2.5 g, 13.6 mmol), BC1₃ (1 M in CH₂C1₂, 13.6 ml), ZnCl₂ (2.04 g, 15 mmol), and chloroacetonitrile (1.03 ml, 16.3 mmol) in dry CH₂C1₂. Purification by column chromatography as described for (5) afforded (8) (1.06 g, 30%) as a cystalline product. 'H-NMR (CDC1₃, 250 MHz): δ 6.5-5.5 (broad s, 2 H); 5.9 (s, 1 H); 4.70 (s, 2 H); 3.95 (s, 3 H); 3.80 (s, 3 H); 3.65 (s, 3 H).

Example 4: 4.5.6-Trimethoxy-3-indolinone (9) Compound (9) was synthesized from (8) in the same manner as described for the synthesis of (6) using 2-amino-4,5,6-trimethoxy- -chloroacetophenone (1.05 g, 4.0 mmol), K₂CO₃ (838 mg, 6.1 mmol), a little KI, and dry acetone (50 ml). Purification by column chromatography (eluent: CH₂Cl₂/MeOH: 9/1) afforded (9) (190 mg, 21%) as red crystals.

Example 5: Preparation of 6-mefhoxy-3-benzofuranone

Chloroacetonitrile (3.5 ml, 55.2 mmol) was added dropwise to a stirred solution containing 3-methoxyphenol (5 ml, 46 mmol) and zinc chloride (6.9 g, 50.6 mmol) in anhydrous dioxane (30 ml) at room temperature. The resulting solution was saturated with dry hydrogen chloride gas. After stirring at room temperature overnight, the yellow precipitate was filtered and washed with anhydrous ether (100 ml). The collected precipitate was dissolved in water (80 ml) and heated to reflux for 1 hour. The solution was allowed to cool to approximately 40 °C, and aqueous sodium hydroxide (20% w/v) (7.5 ml) was added. After stirring at that temperature for 30 minutes a pale yellow precipitate had formed. A heterogeneous system was then taken to pH ~ 7 by addition of hydrochloric acid (1 M). The precipitate was filtered, washed with water, and recrystallized from acetone to give the desired compound as a light yellow powder (4.14 g, 54.8%). 'H NMR: δ (ppm): 3.85 (OMe); 4.9 (CH2); 6.50-6.46 (2 H-Phenyl); 7.70-7.66 (1 H-Phenyl).

Example 6: Preparation of 6-methoxy-3-benzothiofuranone or 4-methoxy-3-benzothiofuranone

Chloroacetonitrile (0.62 ml, 9.8 mmol) was added dropwise to a stirred solution containing 3-methoxythiophenol (1 ml, 8.1 mmol) and aluminum chloride (1.19 g, 8.9 mmol) in anhydrous ether (10 ml) at room temperature. The resulting solution was saturated with dry hydrogen chloride gas. After stirring at room temperature overnight, the pale yellow precipitate was filtered and washed with anhydrous ether (30 ml). The collected precipitate was dissolved in water (25 ml) and heated to reflux for 1 hour. After cooling to approximately 40°C, aqueous sodium hydroxide (20% w/v) (2.6 ml) was added. After stirring at that temperature for 30 minutes the solution was then adjusted to pH = 7 by the addition of hydrochloric acid (1 M). The resulting solution was extracted with ethyl acetate (2 X 50 ml). The combined organic extracts were washed with brine, dried over magnesium sulfate, and concentrated in vacuo to afford an orange oil. Column chromatography, using dichloromethane with 1% methanol, gave a light yellow solid (770 mg, 52.8%), a single compound by TLC and HPLC.

Example 7: Preparation of 4.6-dimethoxy-3-benzofuranone

Benzofuranone (3 g, 18.1 mmol) was dissolved in DMF (100 ml). To this was added Li₂CO₃ (5.4 g, 72.4 mmol) and methyl iodide (3.5 ml, 54.3 mmol) in one portion. A nitrogen atmosphere was maintained, and the reaction was stirred for 18 hours at 70°C. The mixture was filtered, and water was added to the solution. The DMF/water phase was extracted with dichloromethane (3 X 100 ml) and the organic phase was washed with a saturated NaHCO₃ solution (2 X 100 ml). After drying with MgSO₄ and concentrating the solution, solid yellow crystals formed. These were washed with cold ethanol to give a single compound by HPLC. Yield: 2.2 g (73%). Example 8: Preparation of 4.6-dimethoxy-3-benzofuranthione

This reaction was performed under nitrogen and anhydrous conditions. 4,6-Dimethoxy-3-benzofuranone (1.5 g, 6.6 mmol) was dissolved in dry toluene (25 ml), and Lawesson's reagent (1.6 g, 4 mmol) was added. The mixture was refluxed with stirring for 18 hours. The mixture was cooled to room temperature and purified by chromatography with 1 :1 etheπpetroleum ether. Concentration of the fractions yielded yellow/orange crystals. The crystals were washed with the eluent and clean yellow crystals were produced. Η NMR showed at least two compounds, probably the thioketone and the thiol. Yield: 400 mg (27%). NMR: δ(ppm): 3.86, 3.88, 3.94, 3.96 (OMe, 2 from thioketone and 2 from thiol), δ (ppm): 4.04 (-OCH₂C(S)-), δ (ppm): 6.40 6.52 (aromatic).

Example 9: 4-Methoxy-3 -benzofuranone (2) and 6-methoxy-3-benzofuranone (3) These compounds were prepared from 3-methoxyphenol (1) (6 ml, 55 mmol), ZnCl₂ (8.2 g, 60 mmol), and chloroacetonitrile (4.2 g, 66 mmol) in dry ether (100 ml). Purification of the crude product by column chromatography (eluent:

CH₂Cl₂/MeOH: 99/1) gave (2) (1.147 g, 13%), a single compound by HPLC (98% pure, recrystallized from EtOH). 'H-NMR (Acetone-d₆, 400 MHz): δ( 7.45 (1 H, d); 6.64 (2 H, m); 4.62 (s, 2 H); 3.95 (s, 3 H) and the more polar compound (3) (1.48 g, 17%). HPLC (84% purity). 'H-NMR (Acetone -d₆): δ(400 MHz): values corresponded to previous synthesis of (3).

Example 10: 4.6-Bismethoxymethoxybenzofuranone

To two grams (12 mmol) benzofuranone dissolved in 100 ml DMF was added 6 ml triethylamine. After adding 2.8 ml (36 mmol) methoxymethyl chloride (MOM-C1) dropwise over 15 minutes, the solution was stirred for 18 hours at room temperature. Water (100 ml) was added to quench the excess MOM-C1. The resultant mixture was extracted with 100 ml brine, dried with MgSO₄, and concentrated to give a brown oil which contained some DMF. Chromatographic purification (3:1 ethyl acetate:hexane with 2% diisopropylethylamine) gave the di-MOM protected benzofuranone as an oil. Recrystallization from water gave the product as fine, light brown needles (2.6 g, 70%).

Example 11 : 4.6-Bismethoxymethoxybenzofuranone thioketone This reaction was performed under nitrogen and anhydrous conditions. To a solution of the product of Example 12 (0.3 g, 1.35 mmol) dissolved in 15 ml dry toluene was added Lawesson's reagent (0.36 g, 0.9 mmol). The reaction was refluxed overnight until the ketone was consumed, by TLC. Chromatographic purification (3:1 dichloromethane: petroleum ether and 2% diisopropylethylamine; or 3:1 etheπpetroleum ether and 2% diisopropylethylamine) gave a slightly yellow clear oil. TLC showed a major and minor product.

Example 12: Inhibition of Fungal Growth

Each test compound was tested against nine isolates in an eight-point dose response assay ranging from 50 μg/ml to 0.39 μg/ml. Aspergillus MIC's (minimum inhibitory concentrations) were scored visually after 48 and 72 hours at 37 °C. All

Candida MIC's were scored visually after a 24 hour incubation at 35 °C.

Amphotericin B (2.5 μg/ml) and 5-flucytosine (2.0 μg/ml) were standard controls for each antifungal assay. In each case, total inhibition was observed for all assays relative to amphotericin B and 5-flucytosine. The results are shown below in Table 4, MIC values in μg/ml after 72 hours. HFF toxicity was analyzed after a 24 hour incubation at 37°C (5% CO₂ ). MTS/PMS was added, and the sample absorbance was read at 450 nm. TABLE 4

Minimum Inhibitory Concentrations

Pathogen S02 S12 S17

^a A. fumigatis 12.5 12.5 6.25 ATCC8001 (XI)

A. fumigatus 6.25 (>50) 6.25 (50) 6.25 (25) ATCC8001 (X2)

^b A. fumigatus 6.25 (>50) 6.25 (50) 6.25 (>5C 94-2766

^c A. niger >50 ( ;>50) 50 (50) 12.5 (50)

C albicans 12.5 12.5 3.125

ATCC90028

C. tropicalis 12.5 >50 >50 ATCC750

C. krusei 6.25 6.25 6.25 ATCC6258

^d C. glabrata 0.39 0.39 0.39 (Fluconazole resistant)

C. parapsilosis 6.25 6.25 12.5 ATCC90018

^a Reference strain from Chrisope Technologies ^b Clinical isolate from J. R. Graybill ^c Clinical isolate from A. Sugar ^d Clinical isolate from M. Rinaldi

Scheme P-l shows several compounds of the invention. Scheme P-l

Scheme P-l

Sll

S12 S13 Scheme P-l

S15

S18 S19 All publications and patents mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication or patent was specifically and individually indicated to be incorporated by reference.

Other Embodiments From the foregoing description, it will be apparent that variations and modifications may be made to the invention described herein to adopt it to various usages and conditions. Such embodiments are also within the scope of the following claims.

What is claimed is:

Claims

Claims 1. A method for treating a microbial infection, said method comprising administering to a patient a pharmaceutical composition containing a compound of formula (IA):

(IA)

wherein each R is independently H, OH, Br, Cl, I, amino, thiol, nitro, C,.₄ alkoxy, C_M alkenyloxy, C₂.₆ alkoxyalkyleneoxy, C alkylthio, C₃_,₈ alkyl, or C₃_,₈ alkenyl; or two adjacent Rs, taken together, are a C_2-18 bivalent moiety containing at least one oxgen atom, substituted or disubstituted with A or B, or both, A being H, OH, Br, Cl, I, amino, or thiol, and B being H, C,_.10 alkyl, C₂.₁₈ alkenyl, or C₆.₁₈ aryl; provided that at least two Rs are not H; further provided that when each of two Rs is one of OH, C alkoxy, C,.₄ alkenyloxy, or C₂.₆ alkoxyalkyleneoxy, and X is phenyl substituted with two substituents independently selected from OH, alkoxy, and alkenyloxy, the remaining R cannot be prenyl; further provided that when each of two Rs is one of OH, C,.₄ alkoxy, C alkenyloxy, or C₂_₆ alkoxyalkyleneoxy, and X is phenyl substituted with three substituents independently selected from OH, alkoxy, and alkenyloxy, the remaining R cannot be prenyl; further provided that when each of two Rs is one of OH, C_1-4 alkoxy, C,_₄ alkenyloxy, or C₂.₆ alkoxyalkyleneoxy, and X is phenyl substituted with a prenyl substituent and with two additional substituents independently selected from OH, alkoxy, and alkenyloxy, the remaining R cannot be H or OH; further provided that when each of two Rs is one of OH, C alkoxy, C,_₄ alkenyloxy, or C₂.₆ alkoxyalkyleneoxy, and X is phenyl substituted with a substituent containing three rings and with two additional substituents independently selected from OH, alkoxy, and alkenyloxy, the remaining R cannot be prenyl;

X is C₄.₁₀ alkyl, C₄.₂₀ alkenyl, or a C₄_₂₀ single, C₆.₂₀ bridged, or C₆.₂₀ fused ring moiety containing cycloalkyl, cycloalkenyl, aryl, heterocycle, or heteroaryl, wherein X is substituted with H, OH, Cl, Br, I, amino, cyano, nitro, alkyl, alkoxy, alkenyl, or alkenyloxy; provided that if X is a heteroaryl or heterocyclic moiety where two Rs are each OH and meta to each other, then the remaining R is H and ortho to each of the two hydroxyls, Y and Z are each O, and a ring atom of X is linked directly to the sp² carbon atom adjacent to X, and substituted with H, OH, Cl, Br, I, amino, cyano, alkyl, alkoxy, alkenyl, or alkenyloxy; and each of Y and Z is independently selected from O, S, and NH; or a pharmaceutically acceptable salt or ester thereof.

2. The method of claim 1, wherein X is selected from benzyl, 2,5-dimethoxyphenyl, 2,3-dimethyl-4-methoxyphenyl, 3-benzyloxyphenyl, 3-phenoxyphenyl, 4-benzyloxy-3-methoxyphenyl, 4-[3-propenoic acid]-phenyl, 2-ethoxy- 1 -naphthyl, 1 -(methylthio)ethyl, DL- 1 -phenylethyl, 4-n-pentyloxyphenyl, 1 -(phenylsulfonyl)-2-pyrrolyl, 4-(3-dimethylaminopropoxy)phenyl, 3-phenylpropyl, 2,4-diethoxy-m-tolulyl, 2,6,6-trimethylcyclohexene-l -methyl, 2,5-dimethoxy-3-tetrahydrofuranyl, 4-methyl-5-imidazolyl, 4-n-pentylphenyl, 2-benzyloxy-4,5-dimethoxyphenyl, 1 -pyrenyl, 3,5-dibenzyloxy-3-methoxyphenyl, 3-methyl-4-methoxyphenyl, 4-n-decyloxyphenyl, 2,4-dimethoxy-3-methylphenyl, t-butyl, 3-(4-t-butylphenoxy)phenyl, 2-n-hexyloxyphenyl, 2-(4-chlorophenylthio)phenyl, cyclopropyl, 2,6-dimethoxy-4-hydroxyphenyl, 4-benzyloxyphenyl, 2-benzyloxyphenyl, 8-hydroxy -l,l,7,7-tetramethyljulolidin-9-yl, 2,3,6,7-tetrahydro-8-hydroxyjulolidin-9-yl, 2-methoxymethyl-l-pyrrolidinyl, 5 -(2-nitrophenyl)furanyl, 1 , 1 -dimethyl-2-hydroxy ethyl, 5 -methylfuranyl,

5-(3-chlorophenyl)furanyl, 2,4-hexadienyl, 5-[3(trifluoromethyl)phenylfuranyl], 4,5-dimethyl-4-pentenyl, imidazolyl, ferrocenyl, 2,6-dimethylhept-5-enyl, 5-[2-(trifluoromethyl)phenyl]furanyl, 5-(hydroxy-2-nitromethyl)furanyl, 2,4-dimethyl-2,6-heptadienyl, 1 -phenylethyl, 5-(2-chlorophenyl)furanyl, benzyl, 5-ethyl-2-furanyl, 5-(4-nitrophenyl)furanyl, pentamethylphenyl, l-(methyldithio)isopropyl, 4-trifluoromethylphenyl, 3-fluoro-4-methoxyphenyl, or the X of a compound of Schemes X-l through X-10, wherein the compounds of Schemes X-l through X-10 have the formulae X-CHO.

3. The method of claim 2, wherein said compound is selected from formulae (I), (II), (III), and (IV):

(HI) (IV) wherein V is a bivalent C_2.,_g moiety containing at least one oxygen atom and substituted with A, B, or both; each of W and is independently selected from the values for A, cyano, nitro, C,.₄ alkoxy, C,.₄ alkenyloxy, C₂.₆ alkyloxyalkyleneoxy, C₂.₇ carboxyalkyloxy, C₇.₁₅ arylalkoxy, and C,.₄alkylthio; R_a is H, C₃._1S alkyl, C₃.|₈ alkenyl, C₅.₁₈ cyclohexenyl, or C₆.₁₈ aryl; and each of R_b and R_c is independently selected from H and C alkyl.

4. The method of claim 3, wherein said compound is of the formula Q=(CHX), wherein Q is selected from the Qs of the compounds of Schemes Q-l through Q-l 1, wherein the compounds of Schemes Q-l through Q-l 1 have the formula Q-H₂.

5. The method of claim 1, wherein said compound has an IC₅₀ of less than 50 micrograms per milliliter against at least one pathogenic strain of Candida or Aspergillus.

6. The method of claim 3, wherein V contains between 1 and 3 rings.

7. The method of claim 3, wherein V is selected from the following formulae:

8. The method of claim 3, wherein V is selected from the following formulae:

9. The method of claim 3, wherein R_;, is selected from H, prop-2-enyl. cinnamyl, 2-methylprop-2-enyl, but-2-enyl, 3-methylbut-2-enyl, 3,7-dimethylocta-2,6-dienyl, (cyclohexenyl)methyl, 3,7,1 l-trimethyldodeca-2,6,10-trienyl, and benzyl.

10. The method of claim 3, wherein said compound is of formula (III). and each of Y and Z is independently selected from O and S.

11. The method of claim 1, wherein said compound is selected from S01, S02, S03, S04, S05, S06, S08, S09, S10, Sl l, S12, S13, S14, S15, S16, S17, S18. and S19.

12. The method of claim 2, wherein each of W and W' is independently selected from H, OH, methoxy, methoxymethyleneoxy, and carboxymethoxy.

13. The method of claim 12, wherein Y and Z are O, and at least one of W and W' is OH.

14. The method of claim 2, wherein X is a heterocyclic or heteroaryl moiety.

15. The method of claim 1, wherein X is selected from C₄.₁₀ alkyl, C₄.₂₀ alkenyl, and a C₄.₂₀ single, C₆.₂₀ bridged, or C₆.₂₀ fused ring moiety containing cycloalkyl, cycloalkenyl, or aryl.

16. The method of claim 15, wherein X is a nonaromatic moiety containing cycloalkyl, cycloalkenyl, alkyl, or alkenyl.

17. The method of claim 1, wherein said compound is selected from SI 7 and SI 9.

18. The method of claim 15, wherein X is an aryl moiety.

19. The method of claim 1, wherein said compound is selected from S12 and S02.

20. The method of claim 1, wherein said microbial infection is a fungal infection.

21. The method of claim 20, wherein said fungal infection is an infection of a Candida species.

22. The method of claim 20, wherein said fungal infection is an infection of a fungus resistant to at least one azole antifungal agent.

23. The method of claim 22, wherein said azole antifungal agent is fluconazole.

24. The method of claim 20, wherein said fungal infection is an infection of an Aspergillus species.

25. The method of claim 20, wherein said fungal infection is selected from an infection due to C. albicans, C. glabrata, C krusei, C. tropicalis, C. parapsilosis, A. fumigatus, or A. niger.

26. The method of claim 1, wherein said microbial infection is a bacterial infection.

27. A compound selected from formulae (I)-(IV) below:

(I») wherein V is a bivalent C₂._I8 moiety containing at least one oxygen atom and substituted with A, B, or both; each of W and is independently selected from the values for A, cyano, nitro, C,.₄ alkoxy, C,.₄ alkenyloxy, C₂.₆ alkyloxyalkyleneoxy, C₂.₇ carboxyalkyloxy, C₇.₁₅ arylalkoxy, and C,_-4 alkylthio; R_a is H, C₃.₁₈ alkyl, C₃.₁₈ alkenyl, C₅.₁₈ cyclohexenyl, or C₆_₁₈ aryl; each of R_b and R_c is indpendently selected from H and C,_₄ alkyl;

X is substituted or unsubstituted C₃.₁₅ alkyl, C₃.₁₈ alkenyl, C₃.₁₅ cycloalkyl, C₄_₁₅ cycloalkenyl, C₄_₂₀ bicyclo[a.b.c]alkyl, C₅.₂₀ bicyclo[a.b.c] alkenyl, C₈.₂₀ tricyclo[a.b.c.d]alkyl, C₈.₂₀ tricycloalkenyl, C₂.₂o heterobicyclo[a.b.c]alkyl, or a combination thereof, wherein each of a, b, c, and d is independently 0 to 10; and each of Y and Z is independently selected from O and S.

28. The compound of claim 27, wherein X is C₃.₁₅ alkyl, C₃.₁₈ alkenyl, C₃„ι₅ cycloalkyl, C₄.₁₅ cycloalkenyl, C₅.₁₀ bicyclo[a.b.c]alkyl, C₅.₁₀ bicyclo[a.b.c]alkenyl, C₈.₂₀ tricyclo[a.b.c.d]alkyl, C₈.₂₀ tricycloalkenyl, C₃.₁₀ heterobicyclo[a.b.c]alkyl, or a combination thereof, wherein each of a, b, c, and d is independently 0 to 6.

29. The compound of claim 27, wherein X is C₃.₁₅ alkyl, C₃.₁₈ alkenyl, C₃_ι₅ cycloalkyl, or C₄.₁₅ cycloalkenyl.

30. The compound of claim 27, wherein X is C₅.₁₀ bicyclo[a.b.c.]alkyl,

C_5.10 bicyclo[a.b.c] alkenyl, C₈.₁₅ tricyclo[a.b.c.d]alkyl, C₈.₁₅ tricycloalkenyl, or C_3.10 heterobicyclo[a.b.c]alkyl, or a combination thereof.

31. The compound of claim 28, wherein each of W and W' is independently selected from H, hydroxyl, methoxy, hydroxymethyl, and halomethyl.

32. The compound of claim 28, wherein W and W' are both hydroxyl.