Bicyclic thiophene derivatives and combinatorial libraries thereof

Abstract

The present invention relates to novel bicyclic thiophene derivative compounds of the following formula: 1

Description

BACKGROUND OF THE INVENTION

[0001] 1. Field of the Invention

[0002] The present invention relates generally to the synthesis of compounds comprising heterocyclic rings. In one embodiment, the invention provides novel bicyclic thiophene derivative compounds as well as novel combinatorial libraries comprised of such compounds.

[0003] 2. Background Information

[0004] The process of discovering new therapeutically active compounds for a given indication involves the screening of all compounds from available compound collections. From the compounds tested, one or more structures are selected as a promising lead. A large number of related analogs are then synthesized in order to develop a structure-activity relationship and select one or more optimal compounds. With traditional “one-at-a-time” synthesis and biological testing of analogs, this optimization process is long and labor intensive. Adding significant numbers of new structures to the compound collections used in the initial screening step of the discovery and optimization process cannot be accomplished with traditional “one-at-a-time” synthesis methods, except over a time frame of years or even decades. Faster methods are needed that allow for the preparation of up to thousands of related compounds in a matter of days or a few weeks. This need is particularly evident when it comes to synthesizing more complex compounds, such as bicyclic thiophene derivative compounds.

[0005] Combinatorial approaches have been extended to “organic,” or non-peptide, libraries. Combinatorial chemical methods have even been extended to thiophene derivative compounds, as described, for example, in Sabnis et al., J. Heterocyclic Chem., 36, 333 (1999).

[0006] However, the libraries to date contain compounds of limited diversity and complexity. Such compounds are particularly limited regarding amino and substituted amino radicals attached to the bicyclic thiophene core.

[0007] A need therefore exists to develop more complex libraries based on heterocyclic medicinal compounds which would need less time and effort in the synthesis and testing required to bring an organic pharmaceutical product to fruition. In short, improved methods for generating therapeutically useful heterocyclic compounds, such as bicyclic thiophene derivatives, are desired.

[0008] Thiophene derivative compounds have been the subject of investigation in a number of different biological areas. For example, thiophene derivatives have been proposed or used as antidepressant, antineurodegenerative, antiautoimmune, anticancer, antiallergenic, antihistaminic, antianaphylactic, antiinflamatory or herbicidal agents. See Steiner et al., BASF A.-G, Germany). Ger. Offen. (2000); Richter et al., Novo Nordisk A/S, Den.; Ontogen Corporation; Richter, Birgith). PCT Int. Appl. (1999); Steiner et al., BASF A.-G, Germany). Ger. Offen. (1998); Kretzschmar et al., (VEB Arzneimittelwerk Dresden, Ger. Dem. Rep.). Ger. (East) (1989); Kretzschmar et al., (VEB Arzneimittelwerk Dresden, Ger. Dem. Rep.). Ger. (East) (1989); Tanaka et al., Etp. Appl. Pharmacol., Kyto Pharm. Univ., Misasagi, Japan. Dig. Dis. Sci. (1989); Takahashi et al., (Nippon Surfactant Kogyo K.K., Japan). Jpn. Kokai Tokkyo Koho (1992); Ubusawa et al., (Kureha Chemical Industry Co., Ltd., Japan). Jpn. Kokai Tokkyo Koho (1986); Nakanishi et al., Res. Lab., Yoshitomi Pharm. Ind., Ltd., Tokyo, Japan. Yakugaku Zasshi (1970); Nakanishi et al., Res. Lab., Yoshitomi Pharm. Ind., Ltd., Tokyo, Japan. Yakugaku Zasshi (1970); and El-Subbagh, H. I., Saudi Pharm. J. (1999).

[0009] Bicyclic thiophene derivatives have been the subject of serial chemical synthesis. See, for example, Sabnis et al., J. Heterocyclic Chem., 36, 333 (1999); Norris, R. K., Heterocyclic Compounts: Thiophene derivatives, Vol 4, Pt. 2; McKibben et al., Tetrahedron Letters 40 (1999); Leistner et al., Rep. Synthesis (1987); Artman et al., (NPS Pharmaceuticals, Inc., USA). PCT Int. Appl. (1999); Sensfuss et al., Heteroat. Chem. (1998); Mkrtchyan et al., Inst. Tonkoi Org. Khim., NAN Armenii, Yerevan, Armenia. Khim.-Farm. Zh. (1998); El-Subbagh, H. I., Saudi Pharm. J. (1997); Aries, Robert, Fr. Demande (1973); Nakanishi et al., Yoshitomi Pharmaceutical Industries, Ltd., Japan. (1972); and Nakanishi et al., Yoshitomi Pharmaceutical Industries, Ltd., Japan (1972).

[0010] However, more complex bicyclic thiophene derivatives, especially those with a nitrogen substitution (i.e., amino, (mononsubstituted)amino or (disubstituted)amino) on the non-thiophene ring, have been difficult to attain even through serial methods.

[0011] This invention satisfies this need and provides related advantages as well. The present invention overcomes the known limitations to classical serial organic synthesis of bicyclic thiophene derivatives, for example, as well as the shortcomings of combinatorial chemistry related to bicyclic thiophene derivatives. The present invention allows for rapid generation of large diverse libraries of complex bicyclic thiophene derivatives as discrete molecules. The present invention can utilize a readily available pool of building blocks that can be incorporated into the various regions of the molecule. Furthermore, the method of making the present invention allows for the use of building blocks that contain a wide range of diverse functionality. Such building blocks can provide combinatorial libraries that consist of large numbers as well as combinatorial libraries that are extremely diverse with respect to the functionality contained within those libraries. The present invention combines the techniques of solid-phase synthesis of bicyclic thiophene derivatives and the general techniques of synthesis of combinatorial libraries to prepare highly diverse new bicyclic thiophene derivative compounds.

SUMMARY OF THE INVENTION

[0012] The present invention relates to novel bicyclic thiophene derivative compounds of the following formula: 2

[0013] wherein X, R1, R2, R3 and R4 and n have the meanings provided herein.

[0014] The invention further relates to combinatorial libraries containing two or more such compounds, as well as methods of preparing bicyclic thiophene derivative compounds.

BRIEF DESCRIPTION OF THE DRAWING

[0015] FIG. 1 shows Scheme 1 for the combinatorial synthesis of bicyclic thiophene derivative compounds, where n is 0 or 1 and the depicted exocyclic nitrogen is attached to a sulfonyl derivative (—S(O2)R2). The reagents and conditions for each step of Scheme 1 are as follows: step (a) HO2CCH2CN1 HOBT (1-hydroxybenzo-triazole), DMF (NN-dimethyl-formamide) and DIC at 0° C. for 24 hours; (b) 1-BOC-4-piperidone or N-4-BOC-aminocyclohexane, sulfur, morpholine and DMF at 60° for 20 hours; (c) R1COCL, DIEA (diisopropylcarbodimide), DCM (dichloromethane) for 48 hours at room temperature; (d) 50% TFA/DCM for 30 min. at room temperature; (e) RCO2H, DIEA, HOBT, DIC, DMF or RSO2Cl, DIEA, DCM, THF for 48 hours at room temperature; and (F) HF for 2 hours at room temperature.

DETAILED DESCRIPTION OF THE INVENTION

[0016] The present invention provides compounds and combinatorial libraries of compounds of the formula: 3

[0017] wherein:

[0018] wherein n is 0 or 1; X is C or N, provided that when X is C, n is 1; and when X is N, n is 0;

[0019] R1 is selected from the group consisting of C1-C20 alkyl, C2 to C12 alkenyl, C1 to C12 substituted alkyl, C2 to C12 substituted alkenyl, C2 to C12 substituted alkynyl, C3 to C10 cycloalkyl, C3 to C8 substituted cycloalkyl, C4 to C13 cycloalkenyl, C4 to C13 substituted cycloalkenyl, heteroaryl, substituted heteroaryl, C7 to C18 phenylalkyl, C7 to C18 substituted phenylalkyl, phenyl, substituted phenyl, naphthyl, substituted naphthyl, cyclic C2 to C7 alkylene, substituted cyclic C2 to C7 alkylene, cyclic C2 to C7 heteroalkylene, substituted cyclic C2 to C7 heteroalkylene, carboxy and protected carboxy phthalamide; and,

[0020] R2 is selected from the group consisting of

[0021] (a) a carbonyl group coupled with a functional group selected from the group consisting of C1-C20 alkyl, C2 to C12 alkenyl, C1 to C12 substituted alkyl, C2 to C12 substituted alkenyl, C2 to C12 substituted alkynyl, C3 to C10 cycloalkyl, C3 to C8 substituted cycloalkyl, C7 to C13 cycloalkenyl, C7 to C13 substituted cycloalkanyl, heteroaryl, substituted heteroaryl, C7 to C18 phenylalkyl, C7 to C18 substituted phenylalkyl, phenyl, substituted phenyl, naphthyl, substituted naphthyl, Cyclic C2 to C7 alkylene, substituted cyclic C2 to C7 alkylene, cyclic C2 to C7 heteroalkylene, substituted cyclic C2 to C7 heteroalkylene, carboxy and protected carboxy; and,

[0022] (b) phenylsulfonyl, substituted phenylsulfonyl, C1 to C10 alkylsulfonyl, C1 to C10 substituted alkylsulfonyl, C3 to C10 cycloalkylsulfonyl, C2 to C10 substituted alkylsulfonyl.

[0023] R3 is hydrogen or together with functional Group R1 forms a C2 to C7 substituted or unsubstituted alkylene group.

[0024] R4 is hydrogen or together with functional Group R2 forms a C2 to C7 substituted or unsubstituted alkylene group.

[0025] Preferably, R1 is selected from the list comprising 4-butoxybenzoyl, benzoyl, 2-furoyl, 2-naphthoyl, 1-adamantanecarbonyl, methoxyacetyl, benzothiophene-2-carbonyl, phenoxyacetyl, 2-chlorobenzoyl, 2,3-dimethylbenzoyl, 4-cyanobenzoyl, cyclobutanecarbonyl, benzyloxyacetyl, propionyl, thiophene-2-carbonyl, 3-phenylpropionyl, 4-ethylbenzoyl, 4-tert-butylbenzoyl, 3-methoxyphenylacetyl, 3-bromobenzoyl, 2,4,5-trifluorobenzoyl, 4-n-propylbenzoyl, p-toluoyl, m-toluoyl, o-anisoyl, m-anisoyl, 3-cyclopentylpropionyl, butyryl, 4-chlorobenzoyl, 4-chlorobutyryl, isobutyryl, 2,6-difluorobenzoyl, crotonyl, 2-ethylbutyryl, 3-chloropivaloyl, isovaleryl, nicotinoyl, 3,5,5-trimethylhexanoyl, cyclohexanecarbonyl, 2-phenylbutyryl, cyclopropanecarbonyl, 2,2-di-n-propylacetyl, pentanoyl, 2,4-difluorobenzoyl, 2-ethoxybenzoyl, 2-(trifluoromethyl)benzoyl, 4-n-hexyloxybenzoyl, diphenylacetyl, cyclopentanecarbonyl, pivaloyl; and,

[0026] Preferably, R2 is selected from the group comprising 4-acetamidobenzoyl, 2-(trifluoromethyl)benzoyl, 3,4-difluorobenzoyl, 1-naphthoyl, 3-acetamidobenzoyl, 4-(trifluoromethyl)benzoyl, 2,4-difluorobenzoyl, 7-methoxybenzofuran-2-carbonyl, 2-furoyl, 2-(methylthio)benzoyl, 2-(n-propylthio)nicotinyl, 3-fluoro-2-methyl-benzoyl, 3,4-difluorohydrocinnamyl, 4-chlorocinnamyl, 4-chloro-o-anisyl, 2-chlorocinnamyl, 3-furoyl, 2,5-dimethylphenylacetyl, propionyl, o-toluyl, 2,3-dimethoxybenzoyl, 2-(methylthio)nicotinyl, isobutyryl, crotonyl, thiophene-3-carboxylyl, 2,4-dimethylbenzoyl, cyclohexanecarboxylyl, benzo[b]thiophene-2-carbonyl, isovaleryl, 1-methylindole-3-carbonyl, 2-chlorobenzoyl, 3-chlorobenzoyl, 3-methylthiophene-2-carbonyl, 2,5-dimethoxybenzoyl, 4-chlorobenzoyl, 6-methylnicotinyl, 2-ethoxynicotinyl, cyclobutanecarbonyl, cyclopentylacetyl, 3,5-dimethylisoxazole-4-carbonyl, 1-naphthalenesulfonyl, 2-naphthalenesulfonyl, benzenesulfonyl, 2,5-dichlorobenzenesulfonyl, 2-mesitylenesulfonyl, 4-fluorobenzenesulfonyl, 4-chlorobenzenesulfonyl, 4-methoxybenzenesulfonyl, 4-tert-butylbenzenesulfonyl, p-toluenesulfonyl, methanesulfonyl, beta-styrene sulfonyl, 2,3,5,6-tetramethylbenzenesulfonyl, (trifluoro-4-methyl) benzenesulphonyl, 2,5-dimethoxybenzenesulfonyl, o-toluenesulfonyl , p-xylene-2-sulfonyl, 4-ethylbenzenesulfonyl, 4-n-propylbenzenesulfonyl, 4-n-amylbenzenesulfonyl , 4-isopropylbenzenesulphonyl, 2-fluorobenzenesulphonyl, 3-fluorobenzenesulphonyl, 4-chloro-2,5-dimethylbenzenesulphonyl, 2-chlorobenzenesulfonyl, 3-chlorobenzenesulfonyl, m-toluenesulfonyl, 3,4-dimethoxybenzenesulfonyl, 2,3-dichlorobenzenesulfonyl, 2-bromobenzenesulfonyl, 4-(n-butoxy)benzenesulfonyl, 5-chloro-1,3-dimethylpyrazole-4-sulphonyl, 3,5-dimethylisoxazole-4-sulfonyl, 2,4-dichlorobenzenesulfonyl, 5-fluoro-2-methylbenzenesulfonyl, 5-chloro-2-methoxybenzenesulfonyl, 6-methoxy-m-toluenesulfonyl, 4-biphenylsulfonyl, 4-n-butylbenzenesulfonyl, 4-acetylbenzenesulfonyl

[0027] The invention also provides methods of preparing bicyclic thiophene derivative compounds and combinatorial libraries. In one method, as shown in FIG. 1, such compounds can be prepared by (1) coupling of cyanoacetic acid to a hydroxymethyl resin; (2) Gewald synthesis of 2-aminothiophene; (3) acylation of the amino group; (4) reacting the resulting component in a De-Boc reaction; (5) acylation followed by sulfonation; and (6) cleave in the presence of gaseous HF.

[0028] When the above-described compounds include one or more chiral centers, the stereochemistry of such chiral centers can independently be in the R or S configuration, or a mixture of the two. The chiral centers can be further designated as R or S or R, S or d, D, l, L or d, l, D, L.

[0029] Regarding the compounds and combinatorial libraries described herein, the suffix “ene” added to any of the described radical terms means that the radical is connected to two parts of the compound (for example, methylene (—CH2—), ethylene (—CH2CH2—), etc.).

[0030] The term “C1 to C20 alkyl” denotes such radicals as methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, amyl, tert-amyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl and the like. Preferred “C1 to C20 alkyl” groups are methyl, ethyl, iso-butyl, sec-butyl and iso-propyl. Similarly, the term “C1 to C20 alkylene” denotes radicals of 1 to 20 carbons connected to two other parts in the compound.

[0031] The term “C4 to C13 alkenyl” denotes such radicals as vinyl, allyl, 2-butenyl, 3-butenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 5-heptenyl, 6-heptenyl, (as well as octenyl, nonenyl, decenyl, undecenyl, dodecenyl radicals attached at any appropriate carbon position and the like) as well as dienes and trienes of straight and branched chains.

[0032] The term “C2 to C12 alkynyl” denotes such radicals as ethanol, propynyl, 2-butynyl, 2-pentynyl, 3-pentynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl, 2-heptynyl, 3-heptynyl, 4-heptynyl, 5-heptynyl (as well as octynyl, nonynyl, decynyl, undecynyl, dodecynyl radicals attached at any appropriate carbon position and the like) as well as di- and tri-ynes of straight and branched chains.

[0033] The terms “C1 to C20 substituted alkyl,” “C2 to C12 substituted alkenyl,” “C2 to C20 substituted alkynyl,” “C1 to C12 substituted alkylene,” “C2 to C20 substituted alkenylene” and “C2 to C12 substituted alkynylene” denote groups that are substituted by one or more, and preferably one or two, of halogen, hydroxy, protected hydroxy, oxo, protected oxo, C3 to C7 cycloalkyl, phenyl, naphthyl, amino, protected amino, (monosubstituted)amino, protected (monosubstituted)amino, (disubstituted)amino, guanidino, protected guanidino, heterocyclic ring, substituted heterocyclic ring, imidazolyl, indolyl, pyrrolidinyl, C1 to C12 alkoxy, C1 to C20 acyl, C1 to C20 acyloxy, nitro, carboxy, protected carboxy, carbamoyl, carboxamide, protected carboxamide, N-(C1 to C2O alkyl)carboxamide, protected N-(C1 to C20 alkyl)carboxamide, N,N-di(C1 to C12 alkyl)carboxamide, cyano, methylsulfonylamino, thiol, C1 to C10 alkylthio or C1 to C10 alkylsulfonyl groups. The substituted groups may be substituted once or more, and preferably once or twice, with the same or with different substituents.

[0034] Examples of the above substituted alkyl groups include the 2-oxo-prop-1-yl, 3-oxo-but-1-yl, cyanomethyl, nitromethyl, hydroxymethyl, tetrahydropyranyloxymethyl, trityloxymethyl, propionyloxymethyl, aminomethyl, carboxymethyl, allyloxycarbonylmethyl, allyloxycarbonylaminomethyl, methoxymethyl, ethoxymethyl, t-butoxymethyl, acetoxymethyl, chloromethyl, bromomethyl, iodomethyl, trifluoromethyl, 6-hydroxyhexyl, 2,4-dichloro(n-butyl), 2-aminopropyl, 1-chloroethyl, 2-chloroethyl, 1-bromoethyl, 2-chloroethyl, 1-fluoroethyl, 2-fluoroethyl, 1-iodoethyl, 2-iodoethyl, 1-chloropropyl, 2-chloropropyl, 3-chloropropyl, 1-bromopropyl, 2-bromopropyl, 3-bromopropyl, 1-fluoropropyl, 2-fluoropropyl, 3-fluoropropyl, 1-iodopropyl, 2-iodopropyl, 3-iodopropyl, 2-aminoethyl, 1-aminoethyl, N-benzoyl-2-aminoethyl, N-acetyl-2-aminoethyl, N-benzoyl-1-aminoethyl, N-acetyl-1-aminoethyl and the like.

[0035] Examples of the above substituted alkenyl groups include styrenyl, 3-chloro-propen-1-yl, 3-chloro-buten-1-yl, 3-methoxy-propen-2-yl, 3-phenyl-buten-2-yl, 1-cyano-buten-3-yl and the like. The geometrical isomerism is not critical, and all geometrical isomers for a given substituted alkenyl can be used.

[0036] Examples of the above substituted alkynyl groups include phenylacetylen-1-yl, 1-phenyl-2-propyn-1-yl and the like.

[0037] The term “oxo” denotes a carbon atom bonded to two additional carbon atoms substituted with an oxygen atom doubly bonded to the carbon atom, thereby forming a ketone moiety.

[0038] The term “protected oxo” denotes a carbon atom bonded to two additional carbon atoms substituted with two alkoxy groups or twice bonded to a substituted diol moiety, thereby forming an acyclic or cyclic ketal moiety.

[0039] The term “C1 to C12 alkoxy” as used herein denotes groups such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, t-butoxy and like groups. A preferred alkoxy is methoxy. The term “C1 to C12 substituted alkoxy” means the alkyl portion of the alkoxy can be substituted in the same manner as in relation to C1 to C12 substituted alkyl. Similarly, the term “C1 to C12 phenylalkoxy” as used herein means “C1 to C12 alkoxy” bonded to a phenyl radical.

[0040] The term “C1 to C12 acyloxy” denotes herein groups such as formyloxy, acetoxy, propionyloxy, butyryloxy, pivaloyloxy, pentanoyloxy, hexanoyloxy, heptanoyloxy, octanoyloxy, nonanoyloxy, decanoyloxy, undecanoyloxy, dodecanoyloxy and the like.

[0041] Similarly, the term “C1 to C12 acyl” encompasses groups such as formyl, acetyl, propionyl, butyryl, pentanoyl, pivaloyl, hexanoyl, heptanoyl, octanoyl, nonanoyl, decanoyl, undecanoyl, dodecanoyl, benzoyl and the like. Preferred acyl groups are acetyl and benzoyl.

[0042] The term “C1 to C12 substituted acyl” denotes the acyl group substituted by one or more, and preferably one or two, halogen, hydroxy, protected hydroxy, oxo, protected oxo, cyclohexyl, naphthyl, amino, protected amino, (monosubstituted)amino, protected (monosubstituted)aamino, (disubstituted)amino, guanidino, heterocyclic ring, substituted heterocyclic ring, imidazolyl, indolyl, pyrrolidinyl, C1 to C12 alkoxy, C1 to C12 acyl, C1 to C12 acyloxy, nitro, C1 to C12 alkyl ester, carboxy, protected carboxy, carbamoyl, carboxamide, protected carboxamide, N-(C1 to C12 alkyl)carboxamide, protected N-(C1 to C12 alkyl)carboxamide, N,N-di(C1 to C12 alkyl)carboxamide, cyano, methylsulfonylamino, thiol, C1 to C10 alkylthio or C1 to C10 alkylsulfonyl groups. The substituted acyl groups may be substituted once or more, and preferably once or twice, with the same or with different substituents as described.

[0043] Examples of C1 to C12 substituted acyl groups include 4-phenylbutyroyl, 3-phenylbutyroyl, 3-phenylpropanoyl, 2-cyclohexanylacetyl, cyclohexanecarbonyl, 2-furanoyl and 3-dimethylaminobenzoyl.

[0044] The substituent term “C3 to C7 cycloalkyl” includes the cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl rings. Similarly, a substituent that can be C3 to C7 cycloalkyl” can also be “C5 to C7 cycloalkyl,” which includes the cyclopentyl, cyclohexyl or cycloheptyl rings.

[0045] The substituent term “C3 to C7 substituted cycloalkyl” or “C5 to C7 substituted cycloalkyl” indicates the above cycloalkyl rings substituted by one or two halogen, hydroxy, protected hydroxy, C1 to C10 alkylthio, C1 to C10 alkylsulfoxide, C1 to C10 alkylsulfonyl, C1 to C10 substituted alkylthio, C1 to C10 substituted alkylsulfoxide, C1 to C10 substituted alkylsulfonyl, C1 to C12 alkyl, C1 to C12 alkoxy, C1 to C12 substituted alkyl, C1 to C12 alkoxy, oxo, protected oxo, (monosubstituted)amino, (disubstituted)amino, trifluoromethyl, carboxy, protected carboxy, phenyl, substituted phenyl, phenylthio, phenylsulfoxide, phenylsulfonyl, amino, or protected amino groups.

[0046] The term “cycloalkylene” means a cycloalkyl, as defined above, where the cycloalkyl radical is bonded at two positions connecting together two separate additional groups. Similarly, the term “substituted cycloalkylene” means a cycloalkylene where the cycloalkyl radical is bonded at two positions connecting together two separate additional groups and further bearing at least one additional substituent.

[0047] The term “C5 to C7 cycloalkenyl” indicates a 1,2, or 3-cyclopentenyl ring, a 1,2,3 or 4-cyclohexenyl ring or a 1,2,3,4 or 5-cycloheptenyl ring, while the term “substituted C5 to C7 cycloalkenyl” denotes the above C5 to C7 cycloalkenyl rings substituted by a C1 to C12 alkyl radical, halogen, hydroxy, protected hydroxy, C1 to C12 alkoxy, trifluoromethyl, carboxy, protected carboxy, oxo, protected oxo, (monosubstituted)amino, protected (monosubstituted)amino, (disubstituted)amino, phenyl, substituted phenyl, amino, or protected amino.

[0048] The term “C5 to C7 cycloalkenylene” is a cycloalkenyl ring, as defined above, where the cycloalkenyl radical is bonded at two positions connecting together two separate additional groups. Examples of C5 to C7 cycloalkenylenes include 1,3-cyclopentylene and 1,2-cyclohexylene.

[0049] Similarly, the term “substituted C5 to C7 cycloalkenylene” means a cycloalkenylene further substituted by halogen, hydroxy, protected hydroxy, C1 to C10 alkylthio, C1 to C10 alkylsulfoxide, C1 to C10 alkylsulfonyl, C1 to C10 substituted alkylthio, C1 to C10 substituted alkylsulfoxide, C1 to C10 substituted alkylsulfonyl, C1 to C12 alkyl, C1 to C12 alkoxy, C1 to C12 substituted alkyl, C1 to C12 alkoxy, oxo, protected oxo, (monosubstituted)amino, (disubstituted)amino, trifluoromethyl, carboxy, protected carboxy, phenyl, substituted phenyl, phenylthio, phenylsulfoxide, phenylsulfonyl, amino, or protected amino group. Examples of substituted C5 to C7 cycloalkenylenes include 4-chloro-1,3-cyclopentylene and 4-methyl-1,2-cyclohexylene.

[0050] The term “heterocycle” or “heterocyclic ring” denotes optionally substituted five-membered to eight-membered rings that have 1 to 4 heteroatoms, such as oxygen, sulfur and/or nitrogen, in particular nitrogen, either alone or in conjunction with sulfur or oxygen ring atoms. These five-membered to eight-membered rings may be saturated, fully unsaturated or partially unsaturated, with fully saturated rings being preferred. Preferred heterocyclic rings include morpholino, piperidinyl, piperazinyl, 2-amino-imidazoyl, tetrahydrofurano, pyrrolo, tetrahydrothiophen-yl, hexylmethyleneimino and heptylmethyleneimino.

[0051] The term “substituted heterocycle” or “substituted heterocyclic ring” means the above-described heterocyclic ring substituted with, for example, one or more, and preferably one or two, substituents which are the same or different, which substituents can be halogen, hydroxy, protected hydroxy, cyano, nitro, C1 to C12 alkyl, C1 to C12 alkoxy, C1 to C12 substituted alkoxy, C1 to C12 acyl, C1 to C12 acyloxy, carboxy, protected carboxy, carboxymethyl, protected carboxymethyl, hydroxymethyl, protected hydroxymethyl, amino, protected amino, (monosubstituted)amino, protected (monosubstituted)amino, (disubstituted)amino carboxamide, protected carboxamide, N-(C1 to C12 alkyl)carboxamide, protected N-(C1 to C12 alkyl)carboxamide, N,N-di(C1 to C12 alkyl)carboxamide, trifluoromethyl, N-((C1 to C12 alkyl)sulfonyl)amino, N-(phenylsulfonyl)amino, heterocycle or substituted heterocycle groups.

[0052] The term “heteroaryl” means a heterocyclic aromatic derivative which is a five-membered or six-membered ring system having from 1 to 4 heteroatoms, such as oxygen, sulfur and/or nitrogen, in particular nitrogen, either alone or in conjunction with sulfur or oxygen ring atoms. Examples of heteroaryls include pyridinyl, pyrimidinyl, and pyrazinyl, pyridazinyl, pyrrolo, furano, oxazolo, isoxazolo, thiazolo and the like.

[0053] The term “substituted heteroaryl” means the above-described heteroaryl is substituted with, for example, one or more, and preferably one or two, substituents which are the same or different which substituents can be halogen, hydroxy, protected hydroxy, cyano, nitro, C1 to C12 alkyl, C1 to C12 alkoxy, C1 to C12 substituted alkoxy, C1 to C12 acyl, C1 to C12 substituted acyl, C1 to C12 acyloxy, carboxy, protected carboxy, carboxymethyl, protected carboxymethyl, hydroxymethyl, protected hydroxymethyl, amino, protected amino, (monosubstituted)amino, protected (monosubstituted)amino, (disubstituted)amino, carboxamide, protected carboxamide, N-(C1 to C12 alkyl)carboxamide, protected N-(C1 to C12 alkyl)carboxamide, N,N-di(C1 to C12 alkyl)carboxamide, trifluoromethyl, N-((C1 to C12 alkyl)sulfonyl)amino or N-(phenylsulfonyl)amino groups.

[0054] The term “C7 to C18 phenylalkyl” denotes a C1 to C12 alkyl group substituted at any position within the alkyl chain by a phenyl. The definition includes groups of the formula: -phenyl-alkyl, -alkyl-phenyl and -alkyl-phenyl-alkyl. Examples of such a group include benzyl, 2-phenylethyl, 3-phenyl(n-propyl), 4-phenylhexyl, 3-phenyl(n-amyl), 3-phenyl(sec-butyl) and the like. Preferred C7 to C18 phenylalkyl groups are any one of the preferred alkyl groups described herein combined with a phenyl group.

[0055] Similarly, the term “C1 to C12 heterocycloalkyl” denotes a C1 to C12 alkyl group substituted at any position within the alkyl chain by a “heterocycle,” as defined herein. The definition includes groups of the formula: -heterocyclic-alkyl, -alkyl-heterocyclic and -alkyl-heterocyclic-alkyl. Examples of such a group include 2-pyridylethyl, 3-pierydyl(n-propyl), 4-furylhexyl, 3-piperazyl(n-amyl), 3-morpholyl(sec-butyl) and the like. Preferred C1 to C12 heterocycloalkyl groups are any one of the preferred alkyl groups described herein combined with any one of the preferred heterocycle groups described herein.

[0056] The terms “C7 to C18 substituted phenylalkyl” and “C1 to C12 substituted heterocycloalkyl” denote a C7 to C18 phenylalkyl group or C1 to C12 heterocycloalkyl substituted (on the alkyl or, where applicable, phenyl or heterocyclic portion) with one or more, and preferably one or two, groups chosen from halogen, hydroxy, protected hydroxy, oxo, protected oxo, amino, protected amino, (monosubstituted)amino, protected (monosubstituted)amino, (disubstituted)amino, guanidino, protected guanidino, heterocyclic ring, substituted heterocyclic ring, C1 to C12 alkyl, C1 to C12 substituted alkyl, C1 to C12 alkoxy, C1 to C12 substituted alkoxy, C1 to C12 acyl, C1 to C12 substituted acyl, C1 to C12 acyloxy, nitro, carboxy, protected carboxy, carbamoyl, carboxamide, protected carboxamide, N-(C1 to C12 alkyl)carboxamide, protected N-(C1 to C12 alkyl)carboxamide, N,N-(C1 to C12 dialkyl)carboxamide, cyano, N-(C1 to C12 alkylsulfonyl)amino, thiol, C1 to C10 alkylthio, C1 to C10 alkylsulfonyl groups; and/or the phenyl group may be substituted with one or more, and preferably one or two, substituents chosen from halogen, hydroxy, protected hydroxy, cyano, nitro, C1 to C12 alkyl, C1 to C12 substituted alkyl, C1 to C12 alkoxy, C1 to C12 substituted alkoxy, C1 to C12 acyl, C1 to C12 substituted acyl, C1 to C12 acyloxy, carboxy, protected carboxy, carboxymethyl, protected carboxymethyl, hydroxymethyl, protected hydroxymethyl, amino, protected amino, (monosubstituted)amino, protected (monosubstituted)amino, (disubstituted)amino, carboxamide, protected carboxamide, N-(C1 to C12 alkyl)carboxamide, protected N-(C1 to C12 alkyl)carboxamide, N,N-di(C1 to C12 alkyl)carboxamide, trifluoromethyl, N-((C1 to C12 alkyl)sulfonyl)amino, N-(phenylsulfonyl)amino, Cyclic C2 to C12 alkylene or a phenyl group, substituted or unsubstituted, for a resulting biphenyl group. The substituted alkyl, phenyl or heterocyclic groups may be substituted with one or more, and preferably one or two, substituents which can be the same or different.

[0057] Examples of the term “C7 to C18 substituted phenylalkyl” include groups such as 2-phenyl-1-chloroethyl, 2-(4-methoxyphenyl)ethyl, 4-(2,6-dihydroxy phenyl)n-hexyl, 2-(5-cyano-3-methoxyphenyl)n-pentyl, 3-(2,6-dimethylphenyl)n-propyl, 4-chloro-3-aminobenzyl, 6-(4-methoxyphenyl)-3-carboxy(n-hexyl), 5-(4-aminomethylphenyl)-3-(aminomethyl)n-pentyl, 5-phenyl-3-oxo-n-pent-1-yl and the like.

[0058] The term “C7 to C18 phenylalkylene” specifies a C7 to C18 phenylalkyl, as defined above, where the phenylalkyl radical is bonded at two different positions connecting together two separate additional groups. The definition includes groups of the formula: -phenyl-alkyl-, -alkyl-phenyl- and -alkyl-phenyl-alkyl-. Substitutions on the phenyl ring can be 1,2, 1,3 or 1,4.

[0059] C7 to C18 phenylalkylenes include, for example, 1,4-toluylene and 1,3-xylylene.

[0060] Similarly, the term “C1 to C12 heterocycloalkylene” specifies a C1 to C12 heterocycloalkyl, as defined above, where the heterocycloalkyl radical is bonded at two different positions connecting together two separate additional groups. The definition includes groups of the formula: -heterocyclic-alkyl-, -alkyl-heterocyclic and -alkyl-heterocyclic-alkyl-.

[0061] The terms “C7 to C18 substituted phenylalkylene” and “C1 to C12 substituted heterocycloalkylene” means a C7 to C18 phenylalkylene or C1 to C12 heterocycloalkylene as defined above that is further substituted by halogen, hydroxy, protected hydroxy, C1 to C10 alkylthio, C1 to C10 alkylsulfoxide, C1 to C10 alkylsulfonyl, C1 to C10 substituted alkylthio, C1 to C10 substituted alkylsulfoxide, C1 to C10 substituted alkylsulfonyl, C1 to C12 alkyl, C1 to C12 alkoxy, C1 to C12 substituted alkyl, C1 to C12 alkoxy, oxo, protected oxo, (monosubstituted)amino, (disubstituted)amino, trifluoromethyl, carboxy, protected carboxy, phenyl, substituted phenyl, phenylthio,phenylsulfoxide, phenylsulfonyl, amino, or protected amino group on the phenyl ring or on the alkyl group.

[0062] The term “substituted phenyl” specifies a phenyl group substituted with one or more, and preferably one or two, moieties chosen from the groups consisting of halogen, hydroxy, protected hydroxy, cyano, nitro, C1 to C12 alkyl, C1 to C12 substituted alkyl, C1 to C12 alkoxy, C1 to C12 substituted alkoxy, C1 to C12 acyl, C1 to C12 substituted acyl, C1 to C12 acyloxy, carboxy, protected carboxy, carboxymethyl, protected carboxymethyl, hydroxymethyl, protected hydroxymethyl, amino, protected amino, (monosubstituted)amino, protected (monosubstituted)amino, (disubstituted)amino, carboxamide, protected carboxamide, N-(C1 to C12 alkyl)carboxamide, protected N-(C1 to C12 alkyl)carboxamide, N,N-di(C1 to C12 alkyl)carboxamide, trifluoromethyl, N-((C1 to C12 alkyl)sulfonyl)amino, N-(phenylsulfonyl)amino or phenyl, wherein the phenyl is substituted or unsubstituted, such that, for example, a biphenyl results.

[0063] Examples of the term “substituted phenyl” includes a mono- or di(halo)phenyl group such as 2, 3 or 4-chlorophenyl, 2,6-dichlorophenyl, 2,5-dichlorophenyl, 3,4-dichlorophenyl, 2, 3 or 4-bromophenyl, 3,4-dibromophenyl, 3-chloro-4-fluorophenyl, 2, 3 or 4-fluorophenyl and the like; a mono or di(hydroxy)phenyl group such as 2, 3 or 4-hydroxyphenyl, 2,4-dihydroxyphenyl, the protected-hydroxy derivatives thereof and the like; a nitrophenyl group such as 2, 3 or 4-nitrophenyl; a cyanophenyl group, for example, 2, 3 or 4-cyanophenyl; a mono- or di(alkyl)phenyl group such as 2, 3 or 4-methylphenyl, 2,4-dimethylphenyl, 2, 3 or 4-(iso-propyl)phenyl, 2, 3 or 4-ethylphenyl, 2, 3 or 4-(n-propyl)phenyl and the like; a mono or di(alkoxyl)phenyl group, for example, 2,6-dimethoxyphenyl, 2, 3 or 4-methoxyphenyl, 2, 3 or 4-ethoxyphenyl, 2, 3 or 4-(isopropoxy)phenyl, 2, 3 or 4-(t-butoxy)phenyl, 3-ethoxy-4-methoxyphenyl and the like; 2, 3 or 4-trifluoromethylphenyl; a mono- or dicarboxyphenyl or (protected carboxy)phenyl group such as 2, 3 or 4-carboxyphenyl or 2,4-di(protected carboxy)phenyl; a mono-or di(hydroxymethyl)phenyl or (protected hydroxymethyl)phenyl such as 2, 3, or 4-(protected hydroxymethyl)phenyl or 3,4-di(hydroxymethyl)phenyl; a mono- or di(aminomethyl)phenyl or (protected aminomethyl)phenyl such as 2, 3 or 4-(aminomethyl)phenyl or 2,4-(protected aminomethyl)phenyl; or a mono- or di(N-(methylsulfonylamino))phenyl such as 2, 3 or 4-(N-(methylsulfonylamino))phenyl. Also, the term “substituted phenyl” represents disubstituted phenyl groups wherein the substituents are different, for example, 3-methyl-4-hydroxyphenyl, 3-chloro-4-hydroxyphenyl, 2-methoxy-4-bromophenyl, 4-ethyl-2-hydroxyphenyl, 3-hydroxy-4-nitrophenyl, 2-hydroxy 4-chlorophenyl and the like.

[0064] The term “phenoxy” denotes a phenyl bonded to an oxygen atom, wherein the binding to the rest of the molecule is through the oxygen atom. The term “substituted phenoxy” specifies a phenoxy group substituted with one or more, and preferably one or two, moieties chosen from the groups consisting of halogen, hydroxy, protected hydroxy, cyano, nitro, C1 to C12 alkyl, C1 to C12 alkoxy, C1 to C12 substituted alkoxy, C1 to C12 acyl, C1 to C12 acyloxy, carboxy, protected carboxy, carboxymethyl, protected carboxymethyl, hydroxymethyl, protected hydroxymethyl, amino, protected amino, (monosubstituted)amino, protected (monosubstituted)amino, (disubstituted)amino, carboxamide, protected carboxamide, N-(C1 to C12 alkyl)carboxamide, protected N-(C1 to C12 alkyl)carboxamide, N,N-di(C1 to C12 alkyl)carboxamide, trifluoromethyl, N-((C1 to C12 alkyl)sulfonyl)amino and N-(phenylsulfonyl)amino.

[0065] Examples of substituted phenoxy include 2-methylphenoxy, 2-ethylphenoxy, 2-propylphenoxy, 2-isopropylphenoxy, 2-sec-butylphenoxy, 2-tert-butylphenoxy, 2-allylphenoxy, 2-propenylphenoxy, 2-cyclopentylphenoxy, 2-fluorophenoxy, 2-(trifluoromethyl)phenoxy, 2-chlorophenoxy, 2-bromophenoxy, 2-methoxyphenoxy, 2-ethoxyphenoxy, 2-isopropoxyphenoxy, 3-methylphenoxy, 3-ethylphenoxy, 3-isopropylphenoxy, 3-tert-butylphenoxy, 3-pentadecylphenoxy, 3-(trifluoromethyl)phenoxy, 3-fluorophenoxy, 3-chlorophenoxy, 3-bromophenoxy, 3-iodophenoxy, 3-methoxyphenoxy, 3-(trifluoromethoxy)phenoxy, 4-methylphenoxy, 4-ethylphenoxy, 4-propylphenoxy, 4-isopropylphenoxy, 4-sec-butylphenoxy, 4-tert-butylphenoxy, 4-tert-amylphenoxy, 4-nonylphenoxy, 4-dodecylphenoxy, 4-cyclopenylphenoxy, 4-(trifluoromethyl)phenoxy, 4-fluorophenoxy, 4-chlorophenoxy, 4-bromophenoxy, 4-iodophenoxy, 4-methoxyphenoxy, 4-(trifluoromethoxy)phenoxy, 4-ethoxyphenoxy, 4-propoxyphenoxy, 4-butoxyphenoxy, 4-hexyloxyphenoxy, 4-heptyloxyphenoxy, 2,3-dimethylphenoxy, 5,6,7,8-tetrahydro-1-naphthoxy, 2,3-dichlorophenoxy, 2,3-dihydro-2,2-dimethyl-7-benzofuranoxy, 2,3-dimethoxyphenoxy, 2,6-dimethylphenoxy, 2,6-diisopropylphenoxy, 2,6-di-sec-butylphenoxy, 2-tert-butyl-6-methylphenoxy, 2,6-di-tert-butylphenoxy, 2-allyl-6-methylphenoxy, 2,6-difluorophenoxy, 2,3-difluorophenoxy, 2,6-dichlorophenoxy, 2,6-dibromophenoxy, 2-fluoro-6-methoxyphenoxy, 2,6-dimethoxyphenoxy, 3,5-dimethylphenoxy, 5-isopropyl-3-methylphenoxy, 3,5-di-tert-butylphenoxy, 3,5-bis(trifluoromethyl)phenoxy, 3,5-difluorophenoxy, 3,5-dichlorophenoxy, 3,5-dimethoxyphenoxy, 3-chloro-5-methoxyphenoxy, 3,4-dimethylphenoxy, 5-indanoxy, 5,6,7,8-tetrahydro-2-naphthoxy, 4-chloro-3-methylphenoxy, 2,4-dimethylphenoxy, 2,5-dimethylphenoxy, 2-isopropyl-5-methylphenoxy, 4-isopropyl-3-methylphenoxy, 5-isopropyl-2-methylphenoxy, 2-tert-butyl-5-methylphenoxy, 2-tert-butyl-4-methylphenoxy, 2,4-di-tert-butylphenoxy, 2,4-di-tert-amylphenoxy, 4-fluoro-2-methylphenoxy, 4-fluoro-3-methylphenoxy, 2-chloro-4-methylphenoxy, 2-chloro-5-methylphenoxy, 4-chloro-2-methylphenoxy, 4-chloro-3-ethylphenoxy, 2-bromo-4-methylphenoxy, 4-iodo-2-methylphenoxy, 2-chloro-5-(trifluoromethyl)phenoxy, 2,4-difluorophenoxy, 2,5-difluorophenoxy, 3,4-difluorophenoxy, 4-chloro-2-fluorophenoxy, 3-chloro-4-fluorophenoxy, 4-chloro-3-fluorophenoxy, 2-bromo-4-fluorophenoxy, 4-bromo-2-fluorophenoxy, 2-bromo-5-fluorophenoxy, 2,4-dichlorophenoxy, 3,4-dichlorophenoxy, 2,5-dichlorophenoxy, 2-bromo-4-chlorophenoxy, 2-chloro-4-fluorophenoxy, 4-bromo-2-chlorophenoxy, 2,4-dibromophenoxy, 2-methoxy-4-methylphenoxy, 4-allyl-2-methylphenoxy, trans-2-ethoxy-5-(1-propenyl)phenoxy, 2-methoxy-4-propenylphenoxy, 3,4-dimethoxyphenoxy, 3-ethoxy-4-methoxyphenoxy, 4-allyl-2,6-dimethoxyphenoxy, 3,4-methylenedioxyphenoxy, 2,3,6-trimethylphenoxy, 2,4-dichloro-3-methylphenoxy, 2,3,4-trifluorophenoxy, 2,3,6-trifluorophenoxy, 2,3,5-trifluorophenoxy, 2,3,4-trichlorophenoxy, 2,3,6-trichlorophenoxy, 2,3,5-trimethylphenoxy, 3,4,5-trimethylphenoxy, 4-chloro-3,5-dimethylphenoxy, 4-bromo-3,5-dimethylphenoxy, 2,4,6-trimethylphenoxy, 2,6-bis(hydroxymethyl)-4-methyl phenoxy, 2,6-di-tert-butyl-4-methylphenoxy, 2,6-di-tert-butyl-4-methoxyphenoxy, 2,4,5-trifluorophenoxy, 2-chloro-3,5-difluorophenoxy, 2,4,6-trichlorophenoxy, 3,4,5-trimethoxyphenoxy, 2,3,5-trichlorophenoxy, 4-bromo-2,6-dimethylphenoxy, 4-bromo-6-chloro-2-methylphenoxy, 2,6-dibromo-4-methyl phenoxy, 2,6-dichloro-4-fluorophenoxy, 2,6-dibromo-4-fluorophenoxy, 2,4,6-tribromophenoxy, 2,4,6-triiodophenoxy, 2-chloro-4,5-dimethylphenoxy, 4-chloro-2-isopropyl-5-methylphenoxy, 2-bromo-4,5-difluorophenoxy, 2,4,5-trichlorophenoxy, 2,3,5,6-tetrafluorophenoxy and the like.

[0066] The term “C7 to C18 substituted phenylalkoxy” denotes a C7 to C18 phenylalkoxy group bonded to the rest of the molecule through the oxygen atom, wherein the phenylalkyl portion is substituted with one or more, and preferably one or two, groups selected from halogen, hydroxy, protected hydroxy, oxo, protected oxo, amino, protected amino, (monosubstituted)amino, protected (monosubstituted)amino, (disubstituted)amino, guanidino, heterocyclic ring, substituted heterocyclic ring, C1 to C12 alkoxy, C1 to C12 acyl, C1 to C12 acyloxy, nitro, carboxy, protected carboxy, carbamoyl, carboxamide, protected carboxamide, N-(C1 to C12 alkyl)carboxamide, protected N-(C1 to C12 alkyl)carboxamide, N,N-(C1 to C12 dialkyl)carboxamide, cyano, N-(C1 to C12 alkylsulfonyl)amino, thiol, C1 to C10 alkylthio, C1 to C10 alkylsulfonyl groups; and/or the phenyl group can be substituted with one or more, and preferably one or two, substituents chosen from halogen, hydroxy, protected hydroxy, cyano, nitro, C1 to C12 alkyl, C1 to C12 alkoxy, C1 to C12 acyl, C1 to C12 acyloxy, carboxy, protected carboxy, carboxymethyl, protected carboxymethyl, hydroxymethyl, protected hydroxymethyl, amino, protected amino, (monosubstituted)amino, protected (monosubstituted)amino, (disubstituted)amino, carboxamide, protected carboxamide, N-(C1 to C12 alkyl)carboxamide, protected N-(C1 to C12 alkyl)carboxamide, N,N-di (C1 to C12 alkyl)carboxamide, trifluoromethyl, N-((C1 to C12 alkyl)sulfonyl)amino, N-(phenylsulfonyl)amino or a phenyl group, substituted or unsubstituted, for a resulting biphenyl group. The substituted alkyl or phenyl groups may be substituted with one or more, and preferably one or two, substituents which can be the same or different.

[0067] Examples of the term “C7 to C18 substituted phenylalkoxy” include groups such as 2-(4-hydroxyphenyl)ethoxy, 4-(4-methoxyphenyl)butoxy, (2R)-3-phenyl-2-amino-propoxy, (2S)-3-phenyl-2-amino-propoxy, 2-indanoxy, 6-phenyl-1-hexanoxy, cinnamyloxy, (+/−)-2-phenyl-1-propoxy, 2,2-dimethyl-3-phenyl-1-propoxy and the like.

[0068] The term “phthalimide” means a cyclic imide which is made from phthalic acid, also called 1,2-benzenedicarboxylic acid. The term “substituted phthalimide” specifies a phthalimide group substituted with one or more, and preferably one or two, moieties chosen from the groups consisting of halogen, hydroxy, protected hydroxy, cyano, nitro, C1 to C12 alkyl, C1 to C12 alkoxy, C1 to C12 substituted alkoxy, C1 to C12 acyl, C1 to C12 acyloxy, carboxy, protected carboxy, carboxymethyl, protected carboxymethyl, hydroxymethyl, protected hydroxymethyl, amino, protected amino, (monosubstituted)amino, protected (monosubstituted)amino, (disubstituted)amino, carboxamide, protected carboxamide, N-(C1 to C12 alkyl)carboxamide, protected N-(C1 to C12 alkyl)carboxamide, N,N-di(C1 to C12 alkyl)carboxamide, trifluoromethyl, N-((C1 to C12 alkyl)sulfonyl)amino and N-(phenylsulfonyl)amino.

[0069] Examples of substituted phthalimides include 4,5-dichlorophthalimido, 3-fluorophthalimido, 4-methoxyphthalimido, 3-methylphthalimido, 4-carboxyphthalimido and the like.

[0070] The term “substituted naphthyl” specifies a naphthyl group substituted with one or more, and preferably one or two, moieties either on the same ring or on different rings chosen from the groups consisting of halogen, hydroxy, protected hydroxy, cyano, nitro, C1 to C6 alkyl, C1 to C7 alkoxy, C1 to C7 acyl, C1 to C7 acyloxy, carboxy, protected carboxy, carboxymethyl, protected carboxymethyl, hydroxymethyl, protected hydroxymethyl, amino, protected amino, (monosubstituted)amino, protected (monosubstituted)amino, (disubstituted)amino, carboxamide, protected carboxamide, N-(C1 to C12 alkyl)carboxamide, protected N-(C1 to C12 alkyl)carboxamide, N, N-di(C1 to C12 alkyl)carboxamide, trifluoromethyl, N-((C1 to C12 alkyl)sulfonyl)amino or N-(phenylsulfonyl)amino.

[0071] Examples of the term “substituted naphthyl” includes a mono or di(halo)naphthyl group such as 1, 2, 3, 4, 5, 6, 7 or 8-chloronaphthyl, 2,6-dichloronaphthyl, 2,5-dichloronaphthyl, 3,4-dichloronaphthyl, 1, 2, 3, 4, 5, 6, 7 or 8-bromonaphthyl, 3,4-dibromonaphthyl, 3-chloro-4-fluoronaphthyl, 1, 2, 3, 4, 5, 6, 7 or 8-fluoronaphthyl and the like; a mono or di(hydroxy)naphthyl group such as 1, 2, 3, 4, 5, 6, 7 or 8-hydroxynaphthyl, 2,4-dihydroxynaphthyl, the protected-hydroxy derivatives thereof and the like; a nitronaphthyl group such as 3- or 4-nitronaphthyl; a cyanonaphthyl group, for example, 1, 2, 3, 4, 5, 6, 7 or 8-cyanonaphthyl; a mono- or di(alkyl)naphthyl group such as 2, 3, 4, 5, 6, 7 or 8-methylnaphthyl, 1,2,4-dimethylnaphthyl, 1, 2, 3, 4, 5, 6, 7 or 8-(isopropyl)naphthyl, 1, 2, 3, 4, 5, 6, 7 or 8-ethylnaphthyl, 1, 2, 3, 4, 5, 6, 7 or 8-(n-propyl)naphthyl and the like; a mono or di(alkoxy)naphthyl group, for example, 2,6-dimethoxynaphthyl, 1, 2, 3, 4, 5, 6, 7 or 8-methoxynaphthyl, 1, 2, 3, 4, 5, 6, 7 or 8-ethoxynaphthyl, 1, 2, 3, 4, 5, 6, 7 or 8-(isopropoxy)naphthyl, 1, 2, 3, 4, 5, 6, 7 or 8-(t-butoxy)naphthyl, 3-ethoxy-4-methoxynaphthyl and the like; 1, 2, 3, 4, 5, 6, 7 or 8-trifluoromethylnaphthyl; a mono- or dicarboxynaphthyl or (protected carboxy)naphthyl group such as 1, 2, 3, 4, 5, 6, 7 or 8-carboxynaphthyl or 2,4-di(-protected carboxy)naphthyl; a mono- or di(hydroxymethyl)naphthyl or (protected hydroxymethyl)naphthyl such as 1, 2, 3, 4, 5, 6, 7 or 8-(protected hydroxymethyl)naphthyl or 3,4-di(hydroxymethyl)naphthyl; a mono- or di(amino)naphthyl or (protected amino)naphthyl such as 1, 2, 3, 4, 5, 6, 7 or 8-(amino)naphthyl or 2,4-(protected amino)-naphthyl, a mono- or di(aminomethyl)naphthyl or (protected aminomethyl)naphthyl such as 2, 3, or 4-(aminomethyl)naphthyl or 2,4-(protected aminomethyl)-naphthyl; or a mono- or di-(N-methylsulfonylamino) naphthyl such as 1, 2, 3, 4, 5, 6, 7 or 8-(N-methylsulfonylamino)naphthyl. Also, the term “substituted naphthyl” represents disubstituted naphthyl groups wherein the substituents are different, for example, 3-methyl-4-hydroxynaphth-1-yl, 3-chloro-4-hydroxynaphth-2-yl, 2-methoxy-4-bromonaphth-1-yl, 4-ethyl-2-hydroxynaphth-1-yl, 3-hydroxy-4-nitronaphth-2-yl, 2-hydroxy-4-chloronaphth-1-yl, 2-methoxy-7-bromonaphth-1-yl, 4-ethyl-5-hydroxynaphth-2-yl, 3-hydroxy-8-nitronaphth-2-yl, 2-hydroxy-5-chloronaphth-1-yl and the like.

[0072] The term “naphthylene” means a naphthyl radical bonded at two positions connecting together two separate additional groups. Similarly, the term “substituted napthylene” means a naphthylene group that is substituted by halogen, hydroxy, protected hydroxy, C1 to C10 alkylthio, C1 to C10 alkylsulfoxide, C1 to C10 alkylsulfonyl, C1 to C10 substituted alkylthio, C1 to C10 substituted alkylsulfoxide, C1 to C10 substituted alkylsulfonyl, C1 to C12 alkyl, C1 to C12 alkoxy, C1 to C12 substituted alkyl, C1 to C12 alkoxy, oxo, protected oxo, (monosubstituted)amino, (disubstituted)amino, trifluoromethyl, carboxy, protected carboxy, phenyl, substituted phenyl, phenylthio, phenylsulfoxide, phenylsulfonyl, amino, or protected amino group.

[0073] The terms “halo” and “halogen” refer to the fluoro, chloro, bromo or iodo atoms. There can be one or more halogens, which are the same or different. Preferred halogens are chloro and fluoro.

[0074] The term “monosubstituted amino” refers to an amino group with one substituent chosen from the group consisting of phenyl, substituted phenyl, C1 to C12 alkyl, C1 to C12 substituted alkyl, C1 to C12 acyl, C1 to C12 substituted acyl, C2 to C12 alkenyl, C2 to C12 substituted alkenyl, C2 to C12 alkynyl, C2 to C12 substituted alkynyl, C7 to C18 phenylalkyl, C7 to C18 substituted phenylalkyl, heterocyclic ring, substituted heterocyclic ring, C3 to C12 heterocycloalkyl and C3 to C12 substituted heterocycloalkyl cycloalkyl and substcycloalkyl. The (monosubstituted)amino can additionally have an amino-protecting group as encompassed by the term “protected (monosubstituted)amino.”

[0075] The term “disubstituted amino” refers to an amino group with two substituents chosen from the group consisting of phenyl, substituted phenyl, C1 to C12 alkyl, C1 to C12 substituted alkyl, C1 to C12 acyl, C2 to C12 alkenyl, C2 to C12 alkynyl, C7 to C18 phenylalkyl, C7 to C18 substituted phenylalkyl, C3 to C12 heterocycloalkyl and C3 to C12 substituted heterocycloalkyl cycloalkyl and substcyloalkyl. The two substituents can be the same or different.

[0076] The term “amino-protecting group” as used herein refers to substituents of the amino group commonly employed to block or protect the amino functionality while reacting other functional groups of the molecule. The term “protected (monosubstituted)amino” means there is an amino-protecting group on the monosubstituted amino nitrogen atom. In addition, the term “protected carboxamide” means there is an amino-protecting group on the carboxamide nitrogen. Similarly, the term “protected N-(C1 to C12 alkyl)carboxamide” means there is an amino-protecting group on the carboxamide nitrogen.

[0077] Examples of such amino-protecting groups include the formyl (“For”) group, the trityl group, the phthalimido group, the trichloroacetyl group, the chloroacetyl, bromoacetyl, and iodoacetyl groups, urethane-type blocking groups, such as t-butoxycarbonyl (“Boc”), 2-(4-biphenylyl)propyl-2-oxycarbonyl (“Bpoc”), 2-phenylpropyl-2-oxycarbonyl (“Poc”), 2-(4-xenyl)isopropoxycarbonyl, 1,1-diphenylethyl-1-oxycarbonyl, 1,1-diphenylpropyl-1-oxycarbonyl, 2-(3,5-dimethoxyphenyl)propyl-2-oxycarbonyl (“Ddz”), 2-(p-toluyl)propyl-2-oxycarbonyl, cyclopentanyloxycarbonyl, 1-methylcyclopentanyloxycarbonyl, cyclohexanyloxy-carbonyl, 1-methylcyclohexanyloxycarbonyl, 2-methylcyclohexanyloxycarbonyl, 2-(4-toluylsulfonyl)-ethoxycarbonyl, 2-(methylsulfonyl)ethoxycarbonyl, 2-(triphenylphosphino)-ethoxycarbonyl, 9-fluorenylmethoxycarbonyl (“Fmoc”), 2-(trimethylsilyl)ethoxycarbonyl, allyloxycarbonyl, 1-(trimethylsilylmethyl)prop-1-enyloxycarbonyl, 5-benzisoxalylmethoxycarbonyl, 4-acetoxybenzyl-oxycarbonyl, 2,2,2-trichloroethoxycarbonyl, 2-ethynyl-2-propoxycarbonyl, cyclopropylmethoxycarbonyl, isobornyloxycarbonyl, 1-piperidyloxycarbonyl, benzyloxycarbonyl (“Cbz”), 4-phenylbenzyloxycarbonyl, 2-methylbenzyloxy-carbonyl, -2,4,5,-tetramethylbenzyloxycarbonyl (“Tmz”), 4-methoxybenzyloxycarbonyl, 4-fluorobenzyloxycarbonyl, 4-chlorobenzyloxycarbonyl, 3-chlorobenzyloxycarbonyl, 2-chlorobenzyloxycarbonyl, 2,4-dichlorobenzyl-oxycarbonyl, 4-bromobenzyloxycarbonyl, 3-bromobenzyloxycarbonyl, 4-nitrobenzyloxy-carbonyl, 4-cyanobenzyloxycarbonyl, 4-(decyloxy)benzyloxycarbonyl and the like; the benzoylmethylsulfonyl group, dithiasuccinoyl (“Dts”), the 2-(nitro)phenylsulfenyl group (“Nps”), the diphenyl-phosphine oxide group and like amino-protecting groups. The species of amino-protecting group employed is not critical so long as the derivatized amino group is stable to the conditions of the subsequent reaction(s) and can be removed at the appropriate point without disrupting the remainder of the compounds. Preferred amino-protecting groups are Boc, Cbz and Fmoc. Further examples of amino-protecting groups embraced by the above term are well known in organic synthesis and the peptide art and are described by, for example, T. W. Greene and P. G. M. Wuts, “Protective Groups in Organic Synthesis,” 2nd ed., John Wiley and Sons, New York, N.Y., 1991, Chapter 7, M. Bodanzsky, “Principles of Peptide Synthesis,” 1st and 2nd revised ed., Springer-Verlag, New York, N.Y., 1984 and 1993, and Stewart and Young, “Solid Phase Peptide Synthesis,” 2nd ed., Pierce Chemical Co., Rockford, Ill., 1984, each of which is incorporated herein by reference. The related term “protected amino” defines an amino group substituted with an amino-protecting group discussed above.

[0078] The term “protected guanidino” as used herein refers to an “amino-protecting group” on one or two of the guanidino nitrogen atoms. Examples of “protected guanidino” groups are described by T. W. Greene and P. G. M. Wuts; M. Bodanzsky; and Stewart and Young, supra.

[0079] The term “carboxy-protecting group” as used herein refers to one of the ester derivatives of the carboxylic acid group commonly employed to block or protect the carboxylic acid group while reactions are carried out on other functional groups on the compound. Examples of such carboxylic acid protecting groups include t-butyl, 4-nitrobenzyl, 4-methoxybenzyl, 3,4-dimethoxybenzyl, 2,4-dimethoxybenzyl, 2,4,6-trimethoxybenzyl, 2,4,6-trimethylbenzyl, pentamethylbenzyl, 3,4-methylenedioxybenzyl, benzhydryl, 4,4′-dimethoxytrityl, 4,4′,4″-trimethoxytrityl, 2-phenylpropyl, trimethylsilyl, t-butyldimethylsilyl, phenacyl, 2,2,2-trichloroethyl, (trimethylsilyl)ethyl, (di(n-butyl)methylsilyl)ethyl, p-toluenesulfonylethyl, 4-nitrobenzylsulfonylethyl, allyl, cinnamyl, 1-(trimethylsilylmethyl)propenyl and like moieties. The species of carboxy-protecting group employed is not critical so long as the derivatized carboxylic acid is stable to the conditions of subsequent reaction(s) and can be removed at the appropriate point without disrupting the remainder of the molecule. Further examples of these groups are found in E. Haslam, “Protective Groups in Organic Chemistry,” J. G. W. McOmie, Ed., Plenum Press, New York, N.Y., 1973, Chapter 5, and T. W. Greene and P. G. M. Wuts, “Protective Groups in Organic Synthesis,” 2nd ed., John Wiley and Sons, New York, N.Y., 1991, Chapter 5, each of which is incorporated herein by reference. A related term is “protected carboxy,” which refers to a carboxy group substituted with one of the above carboxy-protecting groups.

[0080] The term “hydroxy-protecting group” refers to readily cleavable groups bonded to hydroxyl groups, such as the tetrahydropyranyl, 2-methoxypropyl, 1-ethoxyethyl, methoxymethyl, 2-methoxyethoxymethyl, methylthiomethyl, t-butyl, t-amyl, trityl, 4-methoxytrityl, 4,4′-dimethoxytrityl, 4,4′,4″-trimethoxytrityl, benzyl, allyl, trimethylsilyl, (t-butyl)dimethylsilyl, 2,2,2-trichloroethoxycarbonyl groups and the like. The species of hydroxy-protecting groups is not critical so long as the derivatized hydroxyl group is stable to the conditions of subsequent reaction(s) and can be removed at the appropriate point without disrupting the remainder of the molecule. Further examples of hydroxy-protecting groups are described by C. B. Reese and E. Haslam, “Protective Groups in Organic Chemistry,” J. G. W. McOmie, Ed., Plenum Press, New York, N.Y., 1973, Chapters 3 and 4, respectively, and T. W. Greene and P. G. M. Wuts, “Protective Groups in Organic Synthesis,” 2nd ed., John Wiley and Sons, New York, N.Y., 1991, Chapters 2 and 3. Related terms are “protected hydroxy,” and “protected hydroxymethyl” which refer to a hydroxy or hydroxymethyl substituted with one of the above hydroxy-protecting groups.

[0081] The term “C1 to C10 alkylthio” refers to sulfide groups such as methylthio, ethylthio, n-propylthio, isopropylthio, n-butylthio, t-butylthio and like groups.

[0082] The term “C1 to C10 alkylsulfoxide” indicates sulfoxide groups such as methylsulfoxide, ethylsulfoxide, n-propylsulfoxide, isopropylsulfoxide, n-butylsulfoxide, sec-butylsulfoxide and the like. The term “C1 to C10 alkylsulfonyl” encompasses groups such as methylsulfonyl, ethylsulfonyl, n-propylsulfonyl, isopropylsulfonyl, n-butylsulfonyl, t-butylsulfonyl and the like. it should also be understood that the above thio, sulfoxide or sulfonyl groups can be at any point on the alkyl chain (e.g., 2-methylmercaptoethyl).

[0083] The terms “C1 to C10 substituted alkylthio,” “C1 to C10 substituted alkylsulfoxide,” and “C1 to C10 substituted alkylsulfonyl,” denote the C1 to C10 alkyl portion of these groups may be substituted as described above in relation to “substituted alkyl.”

[0084] The terms “phenylthio,” “phenylsulfoxide,” and “phenylsulfonyl” specify a thiol, a sulfoxide, or sulfone, respectively, containing a phenyl group. The terms “substituted phenylthio,” “substituted phenylsulfoxide,” and “substituted phenylsulfonyl” means that the phenyl ring of these groups can be substituted as described above in relation to “substituted phenyl.”

[0085] The term “C1 to C12 alkoxycarbonyl” means a “C1 to C12 alkoxy” group attached to a carbonyl group. The term “C1 to C12 substituted alkoxycarbonyl” denotes a substituted alkoxy bonded to the carbonyl group, which alkoxy may be substituted as described above in relation to “C1 to C12 substituted alkyl.”

[0086] The term “phenylene” means a phenyl group where the phenyl radical is bonded at two positions connecting together two separate additional groups. Examples of “phenylene” include 1,2-phenylene, 1,3-phenylene, and 1,4-phenylene.

[0087] The term “substituted phenylene” means a phenyl group where the phenyl radical is bonded at two positions connecting together two separate additional groups, wherein the phenyl is substituted as described above in relation to “substituted phenyl.”

[0088] The term “substituted C1 to C12 alkylene” means a C1 to C12 alkyl group where the alkyl radical is connected to two additional groups and further bearing (i.e., substituted with) an additional substituent. Examples of a “substituted C1 to C12 alkylene” include aminomethylene, 1-(amino)-1,2-ethylene, 2-(amino)-1,2-ethylene, 1-(acetamido)-1,2-ethylene, 2-(acetamido)-1,2-ethylene, 2-hydroxy-1,1-ethylene and 1-(amino)-1,3-propylene.

[0089] The terms “cyclic C2 to C7 alkylene,” “substituted cyclic C2 to C7 alkylene,” “cyclic C2 to C7 heteroalkylene,” and “substituted cyclic C2 to C7 heteroalkylene,” defines such a cyclic group bonded (“fused”) to the phenyl radical resulting in a bicyclic ring system. The cyclic group may be saturated or contain one or two double bonds. Furthermore, the cyclic group may have one or two methylene or methine groups replaced by one or two oxygen, nitrogen or sulfur atoms which are the cyclic C2 to C7 heteroalkylene.

[0090] The cyclic alkylene or heteroalkylene group may be substituted once or twice by the same or different substituents which, if appropriate, can be connected to another part of the compound (e.g., alkylene) selected from the group consisting of the following moieties: hydroxy, protected hydroxy, carboxy, protected carboxy, oxo, protected oxo, C1 to C4 acyloxy, formyl, C1 to C12 acyl, C1 to C12 alkyl, C1 to C7 alkoxy, C1 to C10 alkylthio, C1 to C10 alkylsulfoxide, C1 to C10 alkylsulfonyl, halo, amino, protected amino, (monosubstituted)amino, protected (monosubstituted)amino, (disubstituted)amino, hydroxymethyl or a protected hydroxymethyl.

[0091] The cyclic alkylene or heteroalkylene group fused to the benzene radical can contain two to ten ring members and, preferably, contains three to six members. Examples of such saturated cyclic groups are when the resulting bicyclic ring system is a 2,3-dihydro-indanyl or tetralin ring. When the cyclic groups are unsaturated, the resulting bicyclic ring system can be naphthyl or indolyl. Examples of fused cyclic groups which contain one nitrogen atom and one or more double bonds, preferably one or two double bonds, are when the benzene radical is fused to a pyridino, pyrano, pyrrolo, pyridinyl, dihydropyrrolo, or dihydropyridinyl ring. Examples of fused cyclic groups which contain one oxygen atom and one or two double bonds are when the benzene radical ring is fused to a furo, pyrano, dihydrofurano, or dihydropyrano ring. Examples of fused cyclic groups which have one sulfur atom and contain one or two double bonds are when the benzene radical is fused to a thieno, thiopyrano, dihydrothieno or dihydrothiopyrano ring. Examples of cyclic groups which contain two heteroatoms selected from sulfur and nitrogen and one or two double bonds are when the benzene radical ring is fused to a thiazolo, isothiazolo, dihydrothiazolo or dihydroisothiazolo ring. Examples of cyclic groups which contain two heteroatoms selected from oxygen and nitrogen and one or two double bonds are when the benzene ring is fused to an oxazolo, isoxazolo, dihydrooxazolo or dihydroisoxazolo ring. Examples of cyclic groups which contain two nitrogen heteroatoms and one or two double bonds occur when the benzene ring is fused to a pyrazolo, imidazolo, dihydropyrazolo or dihydroimidazolo ring or pyrazinyl.

[0092] One or more of the compounds of the invention, even within a given library, may be present as a salt. The term “salt” encompasses those salts that form with the carboxylate anions and amine nitrogens and include salts formed with the organic and inorganic anions and cations discussed below. Furthermore, the term includes salts that form by standard acid-base reactions with basic groups (such as amino groups) and organic or inorganic acids. Such acids include hydrochloric, hydrofluoric, trifluoroacetic, sulfuric, phosphoric, acetic, succinic, citric, lactic, maleic, fumaric, palmitic, cholic, pamoic, mucic, D-glutamic, D-camphoric, glutaric, phthalic, tartaric, lauric, stearic, salicyclic, methanesulfonic, benzenesulfonic, sorbic, picric, benzoic, cinnamic, and like acids.

[0093] The term “organic or inorganic cation” refers to counter-ions for the carboxylate anion of a carboxylate salt. The counter-ions are chosen from the alkali and alkaline earth metals, (such as lithium, sodium, potassium, barium, aluminum and calcium); ammonium and mono-, di- and tri-alkyl amines such as trimethylamine, cyclohexylamine; and the organic cations, such as dibenzylammonium, benzylammonium, 2-hydroxyethylammonium, bis(2-hydroxyethyl)ammonium, phenylethylbenzylammonium, dibenzylethylenediammonium, and like cations. See, for example, “Pharmaceutical Salts,” Berge et al., J. Pharm. Sci., 66:1-19 (1977), which is incorporated herein by reference. Other cations encompassed by the above term include the protonated form of procaine, quinine and N-methylglucosamine, and the protonated forms of basic amino acids such as glycine, ornithine, histidine, phenylglycine, lysine and arginine. Furthermore, any zwitterionic form of the instant compounds formed by a carboxylic acid and an amino group is referred to by this term. For example, a cation for a carboxylate anion will exist when a position is substituted with a (quaternary ammonium)methyl group. A preferred cation for the carboxylate anion is the sodium cation.

[0094] The compounds of the invention can also exist as solvates and hydrates. Thus, these compounds may crystallize with, for example, waters of hydration, or one, a number of, or any fraction thereof of molecules of the mother liquor solvent. The solvates and hydrates of such compounds are included within the scope of this invention.

[0095] One or more compounds of the invention, even when in a library, can be in the biologically active ester form, such as the non-toxic, metabolically-labile ester-form. Such ester forms induce increased blood levels and prolong the efficacy of the corresponding non-esterified forms of the compounds. Ester groups which can be used include the lower alkoxymethyl groups, for example, methoxymethyl, ethoxymethyl, isopropoxymethyl and the like; the -(C1 to C12) alkoxyethyl groups, for example methoxyethyl, ethoxyethyl, propoxyethyl, isopropoxyethyl and the like; the 2-oxo-1,3-dioxolen-4-ylmethyl groups, such as 5-methyl-2-oxo-1,3-dioxolen-4-ylmethyl, 5-phenyl-2-oxo-1,3-dioxolen-4-ylmethyl and the like; the C1 to C10 alkylthiomethyl groups, for example methylthiomethyl, ethylthiomethyl, iso-propylthiomethyl and the like; the acyloxymethyl groups, for example pivaloyloxymethyl, pivaloyloxyethyl, -acetoxymethyl and the like; the ethoxycarbonyl-1-methyl group; the -acetoxyethyl; the 1-(C1 to C12 alkyloxycarbonyloxy)ethyl groups such as the 1-(ethoxycarbonyloxy)ethyl group; and the 1-(C1 to C12 alkylaminocarbonyloxy)ethyl groups such as the 1-(methylaminocarbonyloxy)ethyl group.

[0096] The term “functionalized resin” means any resin, crosslinked or otherwise, where functional groups have been introduced into the resin, as is common in the art. Such resins include, for example, those functionalized with amino, alkylhalo, formyl or hydroxy groups. Such resins which can serve as solid supports are well known in the art and include, for example, 4-methylbenzhydrylamine-copoly(styrene-1% divinylbenzene) (MBHA), 4-hydroxymethylphenoxymethyl-copoly(styrene-1% divinylbenzene), 4-oxymethyl-phenyl-acetamido-copoly(stryene-1% divinylbenzene) (Wang), 4-(oxymethyl)-phenylacetamido methyl (Pam), and Tentagel™, from Rapp Polymere Gmbh, trialkoxy-diphenyl-methyl ester-copoly(styrene-1% divinylbenzene) (RINK) all of which are commercially available. Other functionalized resins are known in the art and can be use without departure from the scope of the current invention. Such resins may include those described in Jung, G., Combinatorial Peptide and Nonpeptide Libraries, A Handbook (V C H Verlag, 1996) or Bunin, B. A., The Combinatorial Index (Academic Press, 1998), the disclosures of which are incorporated herein by reference.

[0097] As used herein, a “combinatorial library” is an intentionally created collection of differing molecules which can be prepared by the means provided below or otherwise and screened for biological activity in a variety of formats (e.g., libraries of soluble molecules, libraries of compounds attached to resin beads, silica chips or other solid supports). A “combinatorial library,” as defined above, involves successive rounds of chemical syntheses based on a common starting structure. The combinatorial libraries can be screened in any variety of assays, such as those detailed below as well as others useful for assessing their biological activity. The combinatorial libraries will generally have at least one active compound and are generally prepared such that the compounds are in equimolar quantities.

[0098] Compounds described in previous work that are not taught as part of a collection of compounds or not taught as intended for use as part of such a collection are not part of a “combinatorial library” of the invention. In addition, compounds that are in an unintentional or undesired mixture are not part of a “combinatorial library” of the invention.

[0099] A combinatorial library of the invention can contain two or more of the above-described bicyclic thiophene compounds. The invention further provides a combinatorial library containing three, four or five or more of the above-described compounds. In another embodiment of the invention, a combinatorial library can contain ten or more of the above-described compounds. In yet another embodiment of the invention, a combinatorial library can contain fifty or more of the above-described compounds. If desired, a combinatorial library of the invention can contain 100,000 or more, or even 1,000,000 or more, of the above-described compounds.

[0100] By way of example, the preparation of the combinatorial libraries can use the “split resin approach.” The split resin approach is described by, for example, U.S. Pat. No. 5,010,175 to Rutter, WO PCT 91/19735 to Simon, and Gallop et al., J. Med. Chem., 37:1233-1251 (1994), all of which are incorporated herein by reference.

[0101] For preparing pharmaceutical compositions containing compounds of the invention, inert, pharmaceutically acceptable carriers are used. The pharmaceutical carrier can be either solid or liquid. Solid form preparations include, for example, powders, tablets, dispersible granules, capsules, cachets, and suppositories.

[0102] A solid carrier can be one or more substances which can also act as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, or tablet disintegrating agents; it can also be an encapsulating material.

[0103] In powders, the carrier is generally a finely divided solid which is in a mixture with the finely divided active component. In tablets, the active compound is mixed with the carrier having the necessary binding properties in suitable proportions and compacted in the shape and size desired.

[0104] For preparing pharmaceutical composition in the form of suppositories, a low-melting wax such as a mixture of fatty acid glycerides and cocoa butter is first melted and the active ingredient is dispersed therein by, for example, stirring. The molten homogeneous mixture is then poured into convenient-sized molds and allowed to cool and solidify.

[0105] Powders and tablets preferably contain between about 5% to about 70% by weight of the active ingredient. Suitable carriers include, for example, magnesium carbonate, magnesium stearate, talc, lactose, sugar, pectin, dextrin, starch, tragacanth, methyl cellulose, sodium carboxymethyl cellulose, a low-melting wax, cocoa butter and the like.

[0106] The pharmaceutical compositions can include the formulation of the active compound with encapsulating material as a carrier providing a capsule in which the active component (with or without other carriers) is surrounded by a carrier, which is thus in association with it. In a similar manner, cachets are also included. Tablets, powders, cachets, and capsules can be used as solid dosage forms suitable for oral administration.

[0107] Liquid pharmaceutical compositions include, for example, solutions suitable for oral or parenteral administration, or suspensions, and emulsions suitable for oral administration. Sterile water solutions of the active component or sterile solutions of the active component in solvents comprising water, ethanol, or propylene glycol are examples of liquid compositions suitable for parenteral administration.

[0108] Sterile solutions can be prepared by dissolving the active component in the desired solvent system, and then passing the resulting solution through a membrane filter to sterilize it or, alternatively, by dissolving the sterile compound in a previously sterilized solvent under sterile conditions.

[0109] Aqueous solutions for oral administration can be prepared by dissolving the active compound in water and adding suitable flavorants, coloring agents, stabilizers, and thickening agents as desired. Aqueous suspensions for oral use can be made by dispersing the finely divided active component in water together with a viscous material such as natural or synthetic gums, resins, methyl cellulose, sodium carboxymethyl cellulose, and other suspending agents known to the pharmaceutical formulation art.

[0110] Preferably, the pharmaceutical composition is in unit dosage form. In such form, the composition is divided into unit doses containing appropriate quantities of the active bicyclic thiophene compound. The unit dosage form can be a packaged preparation, the package containing discrete quantities of the preparation, for example, packeted tablets, capsules, and powders in vials or ampules. The unit dosage form can also be a capsule, cachet, or tablet itself, or it can be the appropriate number of any of these packaged forms.

[0111] As pharmaceutical compositions for treating infections, pain, or any other indication the compounds of the present invention are generally in a pharmaceutical composition so as to be administered to a subject at dosage levels of from 0.7 to 7000 mg per day, and preferably 1 to 500 mg per day, for a normal human adult of approximately 70 kg of body weight, this translates into a dosage of from 0.01 to 100 mg/kg of body weight per day. The specific dosages employed, however, can be varied depending upon the requirements of the patient, the severity of the condition being treated, and the activity of the compound being employed. The determination of optimum dosages for a particular situation is within the skill of the art.

[0112] The compounds and combinatorial libraries of the invention can be prepared as set forth in FIG. 1, as described below.

[0113] Variant bicyclic thiophene derivative compounds and combinatorial libraries can be prepared in order to achieve a high level of diversity. For instance, as shown in FIGS. 1, such compounds can be prepared by coupling a hydroxymethyl resin with cyanoactic acid (See Step A, FIG. 1) followed by a Gewald synthesis of 2-aminothiophene (See Step B, FIG. 1). The resulting compound contains a -BOC site and an amino site, both of which can undergo reactions to add to add functional groups. The amino group can undergo acylation with a carboxylic acid solution to add a radical group to the thiophene derivative (See Step C, FIG. 1). Following a de-BOC reaction wherein the -BOC group is removed from the thiophene derivative (See Step D of FIG. 1), the thiophene derivative may undergo sulfonation and couple with a sulfonyl chloride derivative (See Step E FIG. 1). The resulting compound can be cleaved from the resin (See Step F of FIG. 1 and step h of FIG. 2).

[0114] Resin-bound bicyclic thiophene derivative compounds can be cleaved by treating them, for example, with HF. They can also be cleaved with TFA/DCM, provided that TFA sensitive protecting group such as Boc are not used in the synthetic scheme. The compounds can be extracted from the spent resin, for example, with AcOH.

[0115] The nonsupport-bound combinatorial libraries can be screened as single compounds. In addition, the nonsupport-bound combinatorial libraries can be screened as mixtures in solution in assays such as radio-receptor inhibition assays, anti-bacterial assays, anti-fungal assays, calmodulin-dependent phosphodiesterase (CaMPDE) assays and phosphodiesterase (PDE) assays, as described in detail below. Deconvolution of highly active mixtures can then be carried out by iterative or positional scanning methods. These techniques, the iterative approach or the positional scanning approach, can be utilized for finding other active compounds within the combinatorial libraries of the present invention using any one of the below-described assays or others well known in the art.

[0116] The iterative approach is well-known and is set forth in general in Houghten et al., Nature, 354, 84-86 (1991) and Dooley et al., Science, 266, 2019-2022 (1994), both of which are incorporated herein by reference. In the iterative approach, for example, sub-libraries of a molecule having three variable groups are made wherein the first variable is defined. Each of the compounds with the defined variable group is reacted with all of the other possibilities at the other two variable groups. These sub-libraries are each tested to define the identity of the second variable in the sub-library having the highest activity in the screen of choice. A new sub-library with the first two variable positions defined is reacted again with all the other possibilities at the remaining undefined variable position. As before, the identity of the third variable position in the sub-library having the highest activity is determined. If more variables exist, this process is repeated for all variables, yielding the compound with each variable contributing to the highest desired activity in the screening process. Promising compounds from this process can then be synthesized on larger scale in traditional single-compound synthetic methods for further biological investigation.

[0117] The positional-scanning approach has been described for various combinatorial libraries as described, for example, in R. Houghten et al. PCT/US91/08694 and U.S. Pat. No. 5,556,762, both of which are incorporated herein by reference. In the positional scanning approach, sublibraries are made defining only one variable with each set of sublibraries and all possible sublibraries with each single variable defined (and all other possibilities at all of the other variable positions), made and tested. From the instant description one skilled in the art could synthesize combinatorial libraries wherein two fixed positions are defined at a time. From the testing of each single-variable defined combinatorial library, the optimum substituent at that position can be determined, pointing to the optimum or at least a series of compounds having a maximum of the desired biological activity. Thus, the number of sublibraries for compounds with a single position defined will be the number of different substituents desired at that position, and the number of all the compounds in each sublibrary will be the product of the number of substituents at each of the other variables.

[0118] Individual compounds and pharmaceutical compositions containing the compounds, as well as methods of using the same, are included within the scope of the present invention. The compounds of the present invention can be used for a variety of purposes and indications and as medicaments for any such purposes and indications. For example, bicyclic thiophene derivative compounds of the present invention can be used as pesticides, acaricides, receptor agonists or antagonists and antimicrobial agents, including antibacterial or antiviral agents. The libraries can be screened in any variety of melanocortin receptor and related activity assays, such as those detailed below as well as others known in the art. Additionally, the subject compounds can be useful as analgesics. Assays which can be used to test the biological activity of the instant compounds include antimicrobial assays, a competitive enzyme-linked immunoabsorbent assay and radio-receptor assays, as described below.

[0119] The melanocortin (MC) receptors are a group of cell surface proteins that mediate a variety of physiological effects, including regulation of adrenal gland function such as production of the glucocorticoids cortisol and aldosterone; control of melanocyte growth and pigment production; thermoregulation; immunomodulation; and analgesia. Five distinct MC receptors have been cloned and are expressed in a variety of tissues, including melanocytes, adrenal cortex, brain, gut, placenta, skeletal muscle, lung, spleen, thymus, bone marrow, pituitary, gonads and adipose tissue (Tatro, Neuroimmunomodulation 3:259-284 (1996)). Three MC receptors, MCR-1, MCR-3 and MCR-4, are expressed in brain tissue (Xia et al., Neuroreport 6:2193-2196 (1995)).

[0120] A variety of ligands termed melanocortins function as agonists that stimulate the activity of MC receptors. The melanocortins include melanocyte-stimulating hormones (MSH) such as α-MSH, β-MSH and γ-MSH, as well as adrenocorticotropic hormone (ACTH). Individual ligands can bind to multiple MC receptors with differing relative affinities. The variety of ligands and MC receptors with differential tissue-specific expression likely provides the molecular basis for the diverse physiological effects of melanocortins and MC receptors. For example, α-MSH antagonizes the actions of immunological substances such as cytokines and acts to modulate fever, inflammation and immune responses (Catania and Lipton, Annals N. Y. Acad. Sci. 680:412-423 (1993)).

[0121] The role of certain specific MC receptors in some of the physiological effects described above for MC receptors has been elucidated. For example, MCR-1 is involved in pain and inflammation. MCR-1 mRNA is expressed in neutrophils (Catania et al., Peptides 17:675-679 (1996)). The anti-inflammatory agent α-MASH was found to inhibit migration of neutrophils. Thus, the presence of MCR-1 in neutrophils correlates with the anti-inflammatory activity of α-MSH.

[0122] An interesting link of MC receptors to regulation of food intake and obesity has recently been described. The brain MC receptor MCR-4 has been shown to function in the regulation of body weight and food intake. Mice in which MCR-4 has been knocked out exhibit weight gain (Huszar et al., Cell 88:131-141 (1997)). In addition, injection into brain of synthetic peptides that mimic melanocortins and bind to MCR-4 caused suppressed feeding in normal and mutant obese mice (Fan et al., Nature 385:165-168 (1997)). These results indicate that the brain MC receptor MCR-4 functions in regulating food intake and body weight.

[0123] Due to the varied physiological activities of MC receptors, high affinity ligands of MC receptors could be used to exploit the varied physiological responses of MC receptors by functioning as potential therapeutic agents or as lead compounds for the development of therapeutic agents. Furthermore, due to the effect of MC receptors on the activity of various cytokines, high affinity MC receptor ligands could also be used to regulate cytokine activity.

[0124] A variety of assays can be used to identify or characterize MC receptor ligands of the invention. For example, the ability of a bicyclic thiophene derivative compound to compete for binding of a known MC receptor ligand can be used to assess the affinity and specificity of a bicyclic thiophene derivative compound for one or more MC receptors. Any MC receptor ligand can be used so long as the ligand can be labeled with a detectable moiety. The detectable moiety can be, for example, a radiolabel, fluorescent label or chromophore, or any detectable functional moiety so long as the MC receptor ligand exhibits specific MC receptor binding. A particularly useful detectable MC receptor ligand for identifying and characterizing other MC receptor ligands is 125I-HP 467, which has the amino acid sequence Ac-Nle-Gln-His-(p(I)-D-Phe)-Arg-(D-Trp)-Gly-NH2 and is described in Dooley et al., “Melanocortin Receptor Ligands and Methods of Using Same,” U.S. patent application Ser. No. 09/027,108, filed Feb. 20, 1998, which is incorporated herein by reference. HP 467 is a para-iodinated form of HP 228.

[0125] Using assay methods such as those described above, binding kinetics and competition with radiolabeled HP 467 can confirm that bicyclic thiophene derivative compounds of the invention bind to one or more MC receptors. Furthermore, bicyclic thiophene derivative compounds of the invention can exhibit a range of affinities and specificity for various MC receptors.

[0126] The invention provides MC receptor ligands that can bind to several MC receptors with similar affinity. In addition, the invention also provides MC receptor ligands that can be selective for one or more MC receptors. As used herein, the term “selective” means that the affinity of a MC receptor ligand differs between one MC receptor and another by about 10-fold, generally about 20- to 50-fold, and particularly about 100-fold. In some cases, a MC receptor ligand having broad specificity is desired. In other cases, it is desirable to use MC receptor ligands having selectivity for a particular MC receptor. For example, MCR-1 ligands are particularly useful for treating pain and inflammation, whereas MCR-4 ligands are useful for treating obesity. The binding characteristics and specificity of a given MC receptor ligand can be selected based on the particular disease or physiological effect that is desired to be altered.

[0127] Another assay useful for identifying or characterizing MC receptor ligands measures signaling of MC receptors. MC receptors are G protein-coupled receptors that couple to adenylate cyclase and produce cAMP. Therefore, measuring cAMP production in a cell expressing a MC receptor and treated with a MC receptor ligand can be used to assess the function of the MC receptor ligand in activating a MC receptor.

[0128] Ligands for MC-3 that can alter the activity of an MC-3 receptor can be useful for treating sexual dysfunction and other conditions or conditions associated with MC-3 such as inflammation. Other MC-3-associated conditions that can be treated with the MC-3 receptor ligands include disuse deconditioning; organ damage such as organ transplantation or ischemic injury; adverse reactions associated with cancer chemotherapy; diseases such as atherosclerosis that are mediated by free radicals and nitric oxide action; bacterial endotoxic sepsis and related shock; adult respiratory distress syndrome; and autoimmune or other patho-immunogenic diseases or reactions such as allergic reactions or anaphylaxis, rheumatoid arthritis, inflammatory bowel disease, ulcerative colitis, glomerulonephritis, systemic lupus erythematosus, transplant atherosclerosis and parasitic mediated immune dysfunctions such as Chagas's disease.

[0129] The invention further provides a method for treating an MC-3-associated condition in a subject. The term “MC-3-associated condition” includes any condition or condition mediated by MC-3 or can be affected by binding an MC-3 ligand. Such conditions include inflammation and sexual dysfunction.

[0130] The term “sexual dysfunction” herein means any condition that inhibits or impairs normal sexual function, including coitus. However, the term need not be limited to physiological conditions, but may include psychogenic conditions or perceived impairment without a formal diagnosis of pathology.

[0131] In males, sexual dysfunction includes erectile dysfunction. The term “erectile dysfunction” or “impotence” means herein the inability or impaired ability to attain or sustain an erection that would be of satisfactory rigidity for coitus. Sexual dysfunction in males can also include premature ejaculation and priapism, which is a condition of prolonged and sometimes painful erection unrelated to sexual activity, often associated with sickle-cell disease.

[0132] In females, sexual dysfunction includes sexual arousal disorder. The term “sexual arousal disorder” means herein a persistent or recurrent failure to attain or maintain the lubrication-swelling response of sexual excitement until completion of sexual activity. Sexual dysfunction in females can also include inhibited orgasm and dyspareunia, which is painful or difficult coitus. Sexual dysfunction can also be manifested as inhibited sexual desire or inhibited lordosis behavior in animals.

[0133] In addition, the ability of the compounds to inhibit bacterial growth, and therefore be useful to that infection, can be determined by methods well known in the art. Compounds of the present invention can be shown to have antimicrobial activity by the in vitro antimicrobial activity assay described below and, therefore, are useful as antimicrobial agents.

[0134] Moreover, an exemplary in vitro antimicrobial activity assay is described in Blondelle and Houghten, Biochemistry 30:4671-4678 (1991), which is incorporated herein by reference. In brief, Staphylococcus aureus ATCC 29213 (Rockville, Md.) is grown overnight at 37° C. in Mueller-Hinton broth, then re-inoculated and incubated at 37° C. to reach the exponential phase of bacterial growth (i.e., a final bacterial suspension containing 105 to 5×105 colony-forming units/ml). The concentration of cells is established by plating 100 μl of the culture solution using serial dilutions (e.g., 10−2, 10−3 and 10−4) onto solid agar plates. In 96-well tissue culture plates, compounds, individual or in mixtures, are added to the bacterial suspension at concentrations derived from serial two-fold dilutions ranging from 1500 to 2.9 μg/ml. The plates are incubated overnight at 37° C. and the growth determined at each concentration by OD620 nm. The IC50 (the concentration necessary to inhibit 50% of the growth of the bacteria) can then be calculated.

[0135] The competitive ELISA method which can be used here is a modification of the direct ELISA technique described previously in Appel et al., J. Immunol. 144:976-983 (1990), which is incorporated herein by reference. It differs only in the MAb addition step. Briefly, multi-well microplates are coated with the antigenic peptide (Ac-GASPYPNLSNQQT-NH2) at a concentration of 100 pmol/50 μl. After blocking, 25 μl of a 1.0 mg/ml solution of each mixture of a synthetic combinatorial library (or individual compound) is added, followed by MAb 125-10F3 (Appel et al., supra) (25 μl per well). The MAb is added at a fixed dilution in which the bicyclic guanidine in solution effectively competes for MAb binding with the antigenic peptide adsorbed to the plate. The remaining steps are the same as for direct ELISA. The concentration of compound necessary to inhibit 50% of the MAb binding to the control peptide on the plate (IC50) is determined by serial dilutions of the compound.

[0136] Alternative screening can be done with radio-receptor assays. The radio-receptor assay, can be selective for any one of the μ, κ, or δ opiate receptors. Compounds of the present invention can be useful in vitro for the diagnosis of relevant opioid receptor subtypes, such as κ, in the brain and other tissue samples. Similarly, the compounds can be used in vivo diagnostically to localize opioid receptor subtypes.

[0137] The radio-receptor assays are also an indication of the compounds' analgesic properties as described, for example, in Dooley et al., Proc. Natl. Acad. Sci., 90:10811-10815 (1993). For example, it can be envisioned that these compounds can be used for therapeutic purposes to block the peripheral effects of a centrally acting pain killer. For instance, morphine is a centrally acting pain killer. Morphine, however, has a number of deleterious effects in the periphery which are not required for the desired analgesic effects, such as constipation and pruritus (itching). While it is known that the many compounds do not readily cross the blood-brain barrier and, therefore, elicit no central effect, the subject compounds can have value in blocking the periphery effects of morphine, such as constipation and pruritus. Accordingly, the subject compounds can also be useful as drugs, namely as analgesics, or to treat pathologies associated with other compounds which interact with the opioid receptor system.

[0138] Additionally, such compounds can be tested in a a receptor assay. Ligands for the a receptor can be useful as antipsychotic agents, as described in Abou-Gharbia et al., Annual Reports in Medicinal Chemistry, 28:1-10 (1993).

[0139] Radio-receptor assays can be performed with particulate membranes prepared using a modification of the method described in Pasternak et al., Mol. Pharmacol. 11:340-351 (1975), which is incorporated herein by reference. Rat brains frozen in liquid nitrogen can be obtained from Rockland (Gilbertsville, Pa.). The brains are thawed, the cerebella removed and the remaining tissue weighed. Each brain is individually homogenized in 40 ml Tris-HCl buffer (50 mM, pH 7.4, 4° C.) and centrifuged (Sorvall® RC5C SA-600: Du Pont, Wilmington, Del.) (16,000 rpm) for 10 minutes. The pellets are resuspended in fresh Tris-HCl buffer and incubated at 37° C. for 40 minutes. Following incubation, the suspensions are centrifuged as before, the resulting pellets resuspended in 100 volumes of Tris buffer and the suspensions combined. Membrane suspensions are prepared and used in the same day. Protein content of the crude homogenates generally range from 0.15-0.2 mg/ml as determined using the method described in Bradford, M. M., Anal. Biochem. 72:248-254 (1976), which is incorporated herein by reference.

[0140] Binding assays are carried out in polypropylene tubes, each tube containing 0.5 ml of membrane suspension. 8 nM of 3H-[D-Ala2,Me-Phe4,Gly-ol5]enkephalin (DAMGO) (specific activity=36 Ci/mmol, 160,000 cpm per tube; which can be obtained from Multiple Peptide Systems, San Diego, Calif., through NIDA drug distribution program 271-90-7302) and 80 μg/ml of bicyclic guanidine, individual or as a mixture and Tris-HCl buffer in a total volume of 0.65 ml. Assay tubes are incubated for 60 mins. at 25° C. The reaction is terminated by filtration through GF-B filters on a Tomtec harvester (Orange, Conn.). The filters are subsequently washed with 6 ml of Tris-HCl buffer, 4° C. Bound radioactivity is counted on a Pharmacia Biotech Betaplate Liquid Scintillation Counter (Piscataway, N.J.) and expressed in cpm. To determine inter- and intra-assay variation, standard curves in which 3H-DAMGO is incubated in the presence of a range of concentrations of unlabeled DAMGO (0.13-3900 nM) are generally included in each plate of each assay (a 96-well format). Competitive inhibition assays are performed as above using serial dilutions of the bicyclic guanidines, individually or in mixtures. IC50 values (the concentration necessary to inhibit 50% of 3H-DAMGO binding) are then calculated. IC50 values of less than 1000 nM are indicative of highly active opioid compounds which bind to the μ receptor, with particularly active compounds having IC50 values of 100 nM or less and the most active compounds with values of less than 10 nM.

[0141] As opposed to this μ receptor selective assay, which can be carried out using 3H-DAMGCO as radioligand, as described above, assays selective for κ receptors can be carried out using [3H]-U69,593 (3 nM, specific activity 62 Ci/mmol) as radioligand. Assays selective for δ opiate receptors can be carried out using tritiated DSLET ([D-Ser2,D-Leu5]-threonine-enkephalin) as radioligand. Assays selective for the o opiate receptor can use radiolabeled pentazocine as ligand.

[0142] Screening of combinatorial libraries and compounds of the invention can be done with an anti-fungal assay. Compounds of the present invention can be useful for treating fungal infections.

[0143] Screening of combinatorial libraries and compounds of the invention also can be done with a calmodulin-dependent phosphodiesterase (CaMPDE) assay. Compounds of the present invention can be useful as calmodulin antagonists.

[0144] Calmodulin (CaM), which is the major intracellular calcium receptor, is involved in many processes that are crucial to cellular viability. In particular, calmodulin is implicated in calcium-stimulated cell proliferation. Calmodulin antagonists are, therefore, useful for treating conditions associated with increased cell proliferation, for example, cancer. In addition, calmodulin antagonists such as compounds of the subject invention are useful both in vitro and in vivo for identifying the role of calmodulin in other biological processes. The disadvantages of known antagonists such as trifluoperazine and N-(4-aminobutyl)-5-chloro-2-naphthalenesulfonamide (W13) include their non-specificity and toxicity. In contrast, advantages of the combinatorial libraries and compounds of the subject invention as calmodulin antagonists include their reduced flexibility and ability to generate broader conformational space of interactive residues as compared to their linear counterparts.

[0145] An example of an assay that identifies CaM antagonists is a CaMPDE assay. In brief, samples are mixed with 50 μl of assay buffer (360 mM Tris, 360 mM Imidazole, 45 mM Mg(CH3COO)2, pH 7.5) and 10 μl of CaCl2 (4.5 mM) to a final volume of 251 μl. 25 μl of calmodulin stock solution (Boehringer Mannheim; 0.01 μg/μl) is then added and the samples then sit at room temperature for 10 minutes. 14 μl of PDE (Sigma; 2 Units dissolved in 4 ml of water; stock concentration: 0.0005 Units/μl) is then added, followed by 50 μl of 5′-nucleotidase (Sigma; 100 Units dissolved in 10 ml of 10 mM Tris-HCl containing 0.5 mM Mg(CH3COO)2, pH 7.0; stock concentration: 10 Units/ml). The samples are then incubated for 10 minutes at 30° C. 50 μl of adenosine 3′,5′-cyclic monophosphate (cAMP) (20 mM in water at pH 7.0) is added, the samples incubated for 1 hour at 30° C. and then vortexed. 200 μl of trichloroacetic acid (TCA) (55% in water) is added to a 200 μl sample aliquot, which is then vortexed and centrifuged for 10 minutes. 80 μl of the resulting supernatants of each sample is transferred to a 96-well plate, with 2 wells each containing 80 μl of each sample. 80 μl of ammonium molybdate (1.1% in 1.1N H2SO4) is then added to all the wells, and the OD of each were determined at 730 nm, with the values later subtracted to the final OD reading. 16 μl of reducing agent (6 g sodium bisulfite, 0.6 g sodium sulfite and 125 mg of 1-amino-2-naphtol-4-sulfonic acid in 50 ml of water) is then added to one of each sample duplicate and 16 μl of water is added to the other duplicate. After sitting for 1 hour at room temperature, the OD of each well is determined at 730 nm. The percent inhibition of calmodulin activity is then calculated for each sample, using as 0% inhibition a control sample containing all reagents without any test samples and as 100% inhibition a control sample containing test samples and all reagents except calmodulin. In addition, the percent inhibition of phosphodiesterase activity was determined by following a similar protocol as the CaMPDE assay described above, except not adding calmodulin to the sample mixture and calculating the percent inhibition by using as 0% inhibition a control reagent without any test samples and as 100% inhibition a control sample containing test samples and all reagents except cAMP.

[0146] The following examples are provided to illustrate but not limit the present invention. The following abreviations have the corresponding meanings:

[0147] DMF: N,N-dimethylforamide;

[0148] HOBt: 1-hydroxybenzotriazole;

[0149] Boc: tert-butoxycarbonyl;

[0150] DIC: N,N′-diisopropylcarbodiimide;

[0151] TFA: trifluoroacetic acid;

[0152] DIEA: N,N-diisopropylethylamine;

[0153] DCM: dichloromethane;

[0154] RT: room temperature

[0155] MeOH: methanol

EXAMPLE 1

Preparation of Bicyclic Thiophenes

[0156] Step A. Coupling of Cyanoacetic Acid to Hydroxymethyl Resin

[0157] Hydroxymethyl resin (1.8 g., 1.57 mmol OH) was dispensed into a porous polypropylene packet (Tea-bag, 75 mm×80 mm, 65μ). To a Nalgene bottle containing the packet was added DMF (41.4 mL), cyanoacetic acid (1.76 g, 20.7 mmol) and HOBT (2.80 g, 20.7 mmol). The bottle was shaken for 5 minutes and placed in an ice/water bath. DIC (3.16 ml, 20.7 mmol) was added in portions. The reaction mixture was maintained at about 5-10° C. during the addition of DIC. The reaction mixture was then warmed up to room temperature and shaken for 24 h. The packet was washed with DMF (3×80 mL), DCM (3×80 mL) and MeOH (3×80 mL) and dried in air for overnight.

[0158] Step B: Gewald Synthesis of 2-aminothiophene

[0159] The resin-bound cyano acetate in the packet was shaken with a solution of N-4-BOC-aminocyclohexanone (3.33 g, 15.6 mmol mmol), sulfur (0.50 g, 15.6 mmol) and morpholine (1.36 g, 15.6 mmol) in DMF (62 mL) at 60° C. for 20 h. The packet was washed thoroughly with DMF (6×80 mL) and MeOH (3×80 mL), and dried in air for overnight.

[0160] Step C: Acylation of the Amino Group

[0161] To a Nalgene bottle containing the resin-bound 2-aminothiophene in the packet was added DCM (56 mL), DIEA (5.1 mL, 29.5 mmol) and isobutyryl chloride (2.1 g, 19.7 mmol) sequentially. The reaction mixture was then shaken at room temperature for 2 days. The packet was then washed thoroughly with DCM (3×80 mL), DMF (3×80 mL), MeOH (3×80 mL) and dried in air for overnight.

[0162] The following chlorides were coupled with the resin according to the above-described procedures: 4-butoxybenzoyl chloride, benzoyl chloride, 2-furoyl chloride, 2-naphthoyl chloride, 1-adamantanecarbonyl chloride, methoxyacetyl chloride, benzothiophene-2-carbonyl chloride, phenoxyacetyl chloride, 2-chlorobenzoyl chloride, 2,3-dimethylbenzoyl chloride, 4-cyanobenzoyl chloride, cyclobutanecarbonyl chloride, benzyloxyacetyl chloride, propionyl chloride, thiophene-2-carbonyl chloride, 3-phenylpropionyl chloride, 4-ethylbenzoyl chloride, 4-tert-butylbenzoyl chloride, 3-methoxyphenylacetyl chloride, 3-bromobenzoyl chloride, 2,4,5-trifluorobenzoyl chloride, 4-n-propylbenzoyl chloride, p-toluoyl chloride, m-toluoyl chloride, o-anisoyl chloride, m-anisoyl chloride, 3-cyclopentylpropionyl chloride, butyryl chloride, 4-chlorobenzoyl chloride, 4-chlorobutyryl chloride, isobutyryl chloride, 2,6-difluorobenzoyl chloride, crotonyl chloride, 2-ethylbutyryl chloride, 3-chloropivaloyl chloride, isovaleryl chloride, nicotinoyl chloride hydrochloride, 3,5,5-trimethylhexanoyl chloride, cyclohexanecarbonyl chloride, 2-phenylbutyryl chloride, cyclopropanecarbonyl chloride, 2,2-di-n-propylacetyl chloride, pentanoyl chloride, 2,4-difluorobenzoyl chloride, 2-ethoxybenzoyl chloride, 2-(trifluoromethyl)benzoyl chloride, 4-n-hexyloxybenzoyl chloride, diphenylacetyl chloride, cyclopentanecarbonyl chloride, pivaloyl chloride.

[0163] Step D: De-BOC Reaction

[0164] The packet was shaken with 50% TFA/DCM (80 mL) at room temperature for 30 minutes and washed with DCM (3×80 mL), 5% DIEA/DCM (3×80 mL), MeOH (3×80 mL) and dried in air for overnight.

[0165] Step E: Acylation and Sulfonylation

[0166] The resin in the packet was suspended in DMF (20 mL) and the and the resin suspension was distributed equally into 40 wells of a microtiter plate (2 mL×80). The extra DMF was removed from the wells, leaving about 0.2 mL of DMF in each well. An array of 40 different carboxyl acid solutions in DMF (RCOOH, 0.42 M; DIEA, 0.42M; HOBT, 0.36 M) was prepared. To each well was added 1.0 mL of the corresponding carboxylic acid solution and 65 μL of DIC.

[0167] The following carboxylic acids were coupled to the resin according to the above-described procedures: 4-acetamidobenzoic acid, 2-(trifluoromethyl)benzoic acid, 3,4-difluorobenzoic acid, 1-naphthoic acid, 3-acetamidobenzoic acid, 4-(trifluoromethyl)benzoic acid, 2,4-difluorobenzoic acid, 7-methoxybenzofuran-2-carboxylic acid, 2-furoic acid, 2-(methylthio)benzoic acid, 2-(n-propylthio)nicotinic acid, 3-fluoro-2-methyl-benzoic acid, 3,4-difluorohydrocinnamic acid, 4-chlorocinnamic acid, 4-chloro-o-anisic acid, 2-chlorocinnamic acid, 3-furoic acid, 2,5-dimethylphenylacetic acid, propionic acid, o-toluic acid, 2,3-dimethoxybenzoic acid, 2-(methylthio)nicotinic acid, isobutyric acid, crotonic acid, thiophene-3-carboxylic acid, 2,4-dimethylbenzoic acid, cyclohexanecarboxylic acid, benzo[b]thiophene-2-carboxylic acid, isovaleric acid, 1-methylindole-3-carboxylic acid, 2-chiorobenzoic acid, 3-chlorobenzoic acid, 3-methylthiophene-2-carboxylic acid, 2,5-dimethoxybenzoic acid, 4-chlorobenzoic acid, 6-methylnicotinic acid, 2-ethoxynicotinic acid, cyclobutanecarboxylic acid, cyclopentylacetic acid, 3,5-dimethylisoxazole-4-carboxylic acid.

[0168] Alternatively the resin in the packet was distributed equally into 40 wells of a microtiter plate. An array of 40 different sulfonyl chloride solutions in a 2:1 mixture of THF/DCM (RSO2Cl, 0.24M) was prepared to each well was added 125 μL of DIEA and 1.5 mL of the corresponding sulfonyl chloride solution.

[0169] The following sulfonyl chloride solutions were coupled to the resin according to their above-decribed procedures: 1-naphthalenesulfonyl chloride, 2-naphthalenesulfonyl chloride, benzenesulfonyl chloride, 2,5-dichlorobenzenesulfonyl chloride, 2-mesitylenesulfonyl chloride, 4-fluorobenzenesulfonyl chloride, 4-chlorobenzenesulfonyl chloride, 4-methoxybenzenesulfonyl chloride, 4-tert-butylbenzenesulfonyl chloride, p-toluenesulfonyl chloride, methanesulfonyl chloride, beta-styrene sulfonyl chloride, 2,3,5,6-tetramethylbenzenesulfonyl chloride, 3-(trifluoromethyl)benzenesulphonyl chloride, 2,5-dimethoxybenzenesulfonyl chloride, o-toluenesulfonyl chloride, p-xylene-2-sulfonyl chloride, 4-ethylbenzenesulfonyl chloride, 4-n-propylbenzenesulfonyl chloride, 4-n-amylbenzenesulfonyl chloride, 4-isopropylbenzenesulphonyl chloride, 2-fluorobenzenesulphonyl chloride, 3-fluorobenzenesulphonyl chloride, 4-chloro-2,5-dimethylbenzenesulphonyl chloride, 2-chlorobenzenesulfonyl chloride, 3-chlorobenzenesulfonyl chloride, m-toluenesulfonyl chloride, 3,4-dimethoxybenzenesulfonyl chloride, 2,3-dichlorobenzenesulfonyl chloride, 2-bromobenzenesulfonyl chloride, 4-(n-butoxy)benzenesulfonyl chloride, 5-chloro-1,3-dimethylpyrazole-4-sulphonyl chloride, 3,5-dimethylisoxazole-4-sulfonyl chloride, 2,4-dichlorobenzenesulfonyl chloride, 5-fluoro-2-methylbenzenesulfonyl chloride, 5-chloro-2-methoxybenzenesulfonyl chloride, 6-methoxy-m-toluenesulfonyl chloride, 4-biphenylsulfonyl chloride, 4-n-butylbenzenesulfonyl chloride, 4-acetylbenzenesulfonyl chloride.

[0170] The plates were capped tightly and shaken at room temperature for 2 days. The resin was washed with MeOH (3×1 mL/well), DMF 3×1 mL/well), MeOH (6×1 mL/well), and dried in air for three days and then held under vacuum overnight.

[0171] Step F. HF Gas Cleavage

[0172] The plates were treated with gaseous HF at room temperature for 2 hours. The HF was removed completely by a stream of nitrogen and then by vacuum. The resin was extracted with acetic acid (3×0.5 mL/well). The extract was lyophilized to give the thiophene compounds.

EXAMPLE 2

Anti-microbial Screen

[0173] Streptococcus pyogenes (ATCC# 97-03 14289) was grown in Todd Hewitt Broth (THB) (Difco Laboratories #0492-17-6) overnight until reaching an optical density of (OD=0.636 @ 570 nm) by reading 0.1 ml in a 96 well microtiter plate in a Molecular Devices Thermomax. This preparation was kept frozen as stocks in 30% v/v glycerol in 1.5 ml aliquots at −70mC. until use. Prior to experiments, 6 ml aliquots were thawed and diluted into 50 ml 2×THB. 60 ul of this dilution was added to 92 wells of microtiter plate. To three wells THB (200 ul) was added to serve as a blank and a sterility control. Test compounds in DMSO and appropriate concentrations of DMSO were added to Growth/Solvent Controls at 0 time. Plates were read at 0 time at 570 nm in the Molecular Devices plate reader to obtain compounds correction factors for insoluble or colored compounds. Plates were read again at 4 hours.

[0174] Percent inhibition is calculated with the following formulae: 4

[0175] Color correct=O.D. 0 hr−Blank 0 hr)−(Solvent Control 0 hr−Blank 0 hr)

[0176] % Inhibition=100−O.D. test compound 4 hr−Blank 4 hr−color correct O.D. growth/solvent control 4 hr−Blank 4 hr

[0177] Under this formula, the most active compounds tested were as follows:

0.413	99.62732919	0.1776	5	C26H25FN2O4S
0.321	98.89389594	0.1776	6	C24H28N2O4S
0.093	98.7186954	0.1776	7	C20H22N2O4S
0.106	96.85497962	0.1776	8	C24H21N3O5S
0.08	96.64848012	0.1776	9	C22H16N2O5S2
0.064	96.38905067	0.1776	10	C21H19N3O5S
0.071	96.38905067	0.1776	11	C20H16N2O5S
0.097	95.92312172	0.1776	12	C23H21N3O5S
0.304	95.65217391	0.1776	13	C23H26N2O4S
0.139	95.62076749	0.1776	14	C25H32N2O4S2
0.067	94.99126383	0.1776	15	C19H18N2O4S
0.106	94.99126383	0.1776	16	C22H17ClN2O4S
0.228	94.8192322	0.1776	17	C24H22ClN3O4S2
0.068	94.52533489	0.1776	18	C20H16N2O5S
0.181	94.52105852	0.1776	19	C24H19ClF2N2O4S
0.068	93.59347699	0.1776	20	C20H16N2O4S2
0.136	93.12754805	0.1776	21	C24H19ClN2O4S
0.079	93.12754805	0.1776	22	C21H18N2O4S2
0.419	92.66693978	0.1776	23	C29H32N2O4S
0.342	92.33954451	0.1776	24	C26H27N3O5S
0.309	92.01147071	0.1776	25	C26H27N3O4S2
0.523	92.01147071	0.1776	26	C28H29N3O5S
0.077	90.79790332	0.1776	27	C19H20N2O4S
0.074	90.79790332	0.1776	28	C24H21N3O5S
0.084	90.41309431	0.1776	29	C21H18N2O5S
0.178	90.31818182	0.1776	30	C23H19ClN2O5S
0.493	90.02070393	0.1776	31	C26H26N2O4S2
0.224	89.71732896	0.1776	32	C26H25ClN2O4S
0.089	89.40011648	0.1776	33	C20H20N2O4S
0.14	89.40011648	0.1776	34	C28H26N2O5S2
0.79	89.35817805	0.1776	35	C27H26F2N2O4S
0.267	88.5	0.1776	36	C25H25N3O5S
0.344	88.03312629	0.1776	37	C25H22F2N2O4S
0.264	87.60717069	0.1776	38	C26H18F2N2O4S
0.09	87.5364007	0.1776	39	C25HH21N3O4S
0.14	87.07047175	0.1776	40	C22H17ClN2O4S
0.298	86.76771815	0.1776	41	C26H24F2N2O4S
0.253	86.70807453	0.1776	42	C25H22F2N2O4S
0.203	86.60454281	0.1776	43	C24H23N3O4S2
0.291	86.43998361	0.1776	44	C25H24N2O4S2
0.387	86.37681159	0.1776	45	C25H23ClN2O4S
0.317	86.36009353	0.1776	46	C28H24N2O6S
0.1	86.13861386	0.1776	47	C25H24N2O4S
0.282	86.11224908	0.1776	48	C25H21F3N2O4S
0.448	86.0516934	0.1776	49	C25H30N2O6S
0.218	86.04554865	0.1776	50	C25H25N3O4S2
0.243	85.87402162	0.1776	51#NAME?	C20H15ClN2O4S2
0.328	85.78451454	0.1776	52	C26H26N2O4S
0.448	85.12904547	0.1776	53	C26H24F2N2O4S
0.336	84.72049689	0.1776	54	C23H28N2O4S
0.243	84.68132687	0.1776	55	C20H21ClN2O4S
0.192	83.80896913	0.1776	56	C26H20N2O4S
0.299	83.4903728	0.1776	57	C28H31N3O4S
0.413	82.83490373	0.1776	58	C28H30N2O4S
0.364	82.59411107	0.1776	59	C24H19ClF2N2O4S
0.248	82.59411107	0.1776	60	C22H15ClF2N2O4S
0.277	82.59411107	0.1776	61	C22H23ClN2O4S
0.294	82.48663102	0.1776	62	C27H28N2O4S
0.214	82.41118229	0.1776	63	C22H19N3O4S
0.21	82.31818182	0.1776	64	C27H22N2O5S
0.222	82.18442256	0.1776	65	C27H25ClN2O4S
0.31	82.17943466	0.1776	66	C24H24N2O4S
0.198	81.89616253	0.1776	67	C26H34N2O4S
0.205	81.89616253	0.1776	68	C24H36N2O4S
0.16	81.4793244	0.1776	69	C23H19ClN2O5S
0.343	80.86849652	0.1776	70	C27H25F3N2O4S
0.321	80.75201432	0.1776	71	C28H30N2O4S
0.26	80.21302745	0.1776	72	C26H26N2O6S
0.138	80.08153757	0.1776	73	C23H20N2O4S
0.31	79.31420052	0.1776	74	C23H20ClN3O5S
0.401	79.22982384	0.1776	75	C26H32N2O4S
0.109	79.14967967	0.1776	76	C23H20N2O4S2
0.38	79.08902692	0.1776	77	C27H29N3O4S2
0.21	78.87763055	0.1776	78	C27H22N2O4S
0.234	78.41967946	0.1776	79	C23H18ClFN2O4S
0.231	78.12150578	0.1776	80	C20H15ClN2O5S
0.216	77.94232268	0.1776	81	C24H24N2O4S
0.209	77.9188857	0.1776	82	C24H21ClN2O4S
0.475	77.9188857	0.1776	83	C28H30N2O6S
0.255	77.82333209	0.1776	84	C24H21ClN2O6S
0.23	77.66377269	0.1776	85	C24H17ClF2N2O4S
0.166	77.56207675	0.1776	86	C25H31FN2O4S
0.231	77.26341663	0.1776	87	C23H28N2O4S
0.222	77.03235991	0.1776	88	C22H15ClF2N2O4S
0.293	76.69524552	0.1776	89	C26H19ClN2O4S
0.341	76.4389234	0.1776	90	C27H28N2O6S
0.105	76.354106	0.1776	91	C25H20N2O6S
0.28	76.33246366	0.1776	92	C24H20ClN3O5S
0.284	76.06951872	0.1776	93	C24H28N2O4S
0.469	75.95247849	0.1776	94	C26H24F2N2O4S
0.185	75.77272727	0.1776	95	C23H18F2N2O5S
0.258	74.96927489	0.1776	96	C24H21ClN2O4S
0.183	74.95631916	0.1776	97	C24H20F2N2O4S
0.283	74.7826087	0.1776	98	C22H26N2O4S
0.289	74.64154035	0.1776	99	C26H27N3O4S
0.163	74.31818182	0.1776	100	C24H22N2O5S
0.186	73.59090909	0.1776	101	C24H19F3N2O5S
0.158	73.59090909	0.1776	102	C24H21FN2O5S
0.183	73.55853232	0.1776	103	C24H22N2O6S
0.198	73.26578332	0.1776	104	C27H21FN2O4S
0.397	73.00286768	0.1776	105	C30H28N2O4S
0.203	72.87376902	0.1776	106	C25H23ClN2O4S
0.33	72.86096257	0.1776	107	C24H25F3N2O4S
0.249	72.75437943	0.1776	108	C24H20ClN3O5S
0.237	72.75437943	0.1776	109	C22H16Cl2N2O4S
0.195	72.62667443	0.1776	110	C23H17F3N2O4S
0.267	72.51566697	0.1776	111	C24H24N2OS2
0.304	72.45620574	0.1776	112	C22H23ClN2O4S
0.143	72.33047545	0.1776	113	C21H17FN2O5S
0.261	71.85985837	0.1776	114	C23H19ClN2O4S2
0.309	71.69192954	0.1776	115	C28H29N3O5S
0.351	70.66716362	0.1776	116	C24H21ClN2O6S
0.451	69.39778779	0.1776	117	C28H28F2N2O4S
0.257	69.29274843	0.1776	118	C23H26N2O4S
0.304	69.17629519	0.1776	119	C22H18ClN3O4S
0.467	68.91344383	0.1776	120	C28H32N2O6S
0.215	68.89390519	0.1776	121	C28H32N2O4S
0.277	68.74231872	0.1776	122	C25H26N2O4S
0.201	68.57994782	0.1776	123	C24H18Cl2N2O4S
0.166	68.57654432	0.1776	124	C26H26N2O4S2
0.324	68.41458419	0.1776	125	C26H25ClN2O4S
0.21	68.17155756	0.1776	126	C26H34N2O6S
0.247	67.98360045	0.1776	127	C20H19ClN2O4S
0.237	67.75911512	0.1776	128	C25H25N3O5S
0.324	67.51336898	0.1776	129	C25H27ClN2O4S
0.254	67.50223814	0.1776	130	C24H25ClN2O4S
0.211	67.50145603	0.1776	131	C24H18N2O4S2
0.523	67.49482402	0.1776	132	C26H25ClN2O5S
0.351	67.43138058	0.1776	133	C28H24N2O4S
0.273	67.3421668	0.1776	134	C29H26N2O4S
0.244	67.08907939	0.1776	135	C23H19ClN2O4S2
0.286	67.08907939	0.1776	136	C23H16ClF3N2O4S
0.338	67.03039751	0.1776	137	C28H25N3O4S2
0.328	66.83229814	0.1776	138	C25H30N2O4S
0.288	66.7909057	0.1776	139	C24H22ClN3O4S2
0.345	66.78603402	0.1776	140	C24h26F2N2O4S
0.185	66.78603402	0.1776	141	C27H24N2O4S2
0.162	66.71862822	0.1776	142	C18H14N2O5S
0.217	66.40685892	0.1776	143	C26H18F2N2O4S
0.325	66.08734403	0.1776	144	C23H25ClN2O4S
0.243	65.89638464	0.1776	145	C26H19ClN2O4S
0.186	65.78332034	0.1776	146	C23H22N2O4S
0.198	65.78332034	0.1776	147	C20H22N2O5S
0.257	65.63774024	0.1776	148	C22H26N2O5S
0.27	65.37433155	0.1776	149	C23H32N2O4S
0.213	65.13723884	0.1776	150	C22H24N2O4S
0.26	65.01782531	0.1776	151	C26H29N3O4S
0.359	64.78821363	0.1776	152	C27H27F3N2O4S
0.23	64.7036899	0.1776	153	C22H15ClF2N2O4S
0.264	64.53624318	0.1776	154	C28H24N2O6S
0.233	64.53624318	0.1776	155	C26H21N3O4S2
0.421	64.51345756	0.1776	156	C26H23F3N2O4S
0.207	64.5	0.1776	157	C24h22N2O5S
0.252	64.48176977	0.1776	158	C24H30N2O4S
0.286	64.48176977	0.1776	159	C27H28N2O4S
0.321	64.40551621	0.1776	160	C22H18ClN3O4S
0.257	64.40551621	0.1776	161	C23H16ClF3N2O4S
0.286	64.18219462	0.1776	162	C25H23ClN2O4S
0.258	63.92121755	0.1776	163	C23H25FN2O4S
0.202	63.8263007	0.1776	164	C24H23N3O4S
0.212	63.77272727	0.1776	165	C25H20N2O5S2
0.2	63.60093531	0.1776	166	C26H19ClN2O4S
0.37	63.51099515	0.1776	167	C24H21ClN2O4S
0.243	63.49856616	0.1776	168	C24H28N2O4S
0.256	63.47629797	0.1776	169	C24H29ClN2O4S
0.163	63.40909091	0.1776	170	C24H21FN2O5S
0.315	63.28916602	0.1776	171	C27H19F3N2O4S
0.317	63.17083163	0.1776	172	C25H21F3N2O4S
0.212	62.97739673	0.1776	173	C22H20N2O4S
0.244	62.91464778	0.1776	174	C22H16Cl2N2O4S
0.282	62.84309709	0.1776	175	C25H23ClN2Ohd 5S
0.264	62.66562744	0.1776	176	C26H21N3O4S
0.218	62.6164741	0.1776	177	C23H18ClFN2O4S
0.206	62.48880931	0.1776	178	C29H26N2O4S
0.174	62.35385814	0.1776	179	C20H17N3O5S2
0.226	62.31818182	0.1776	180	C26H26N2O5S
0.277	62.18762802	0.1776	181	C27H25N3O4S
0.297	62.02012672	0.1776	182	C21H17ClN2O4S2
0.204	61.95454545	0.1776	183	C26H26N2O5S
0.324	61.84162063	0.1776	184	C27H30N2O4S
0.224	61.59090909	0.1776	185	C25H25N3O5S2
0.382	61.5320911	0.1776	186	C25H30N2O4S
0.518	61.45276292	0.1776	187	C23H24F2N2O4S
0.276	61.20442442	0.1776	188	C24H24N2O5S
0.431	61.20082816	0.1776	189	C27H28N2O4S
0.22	61.2560567	0.1776	190	C22H15ClF2N2O4S
0.298	61.05640107	0.1776	191	C24H28N2O6S
0.236	60.87668988	0.1776	192	C23H22N2O4S2
0.193	60.86363636	0.1776	193	C25H22F2N2O5S
0.219	60.69829902	0.1776	194	C21H24N2O4S
0.318	60.53830228	0.1776	195	C27H28N2O6S
0.23	60.52925829	0.1776	196	C22H18ClN3O4S2
0.231	60.52925829	0.1776	197	C24H21ClN2O6S
0.184	60.4832424	0.1776	198	C20H15ClN2O5S
0.233	59.98421468	0.1776	199	C23H17ClF2N2O5S
0.271	59.89348628	0.1776	200	C26H32N2O4S
0.23	59.85970382	0.1776	201	C24H22N2O4S
0.26	59.66850829	0.1776	202	C23H15F5N2O4S
0.241	59.63473723	0.1776	203	C20H21ClN2O4S
0.433	59.56575174	0.1776	204	C27H25F3N2O4S

EXAMPLE 3

Melanocortin Receptor Assay

[0178] This example describes methods for assaying binding to MC receptors.

[0179] All cell culture media and reagents are obtained from GibcoBRL (Gaithersburg Md.), except for COSMIC CALF SERUM (HyClone; Logan Utah). HEK 293 cell lines are transfected with the human MC receptors hMCR-1, hMCR-3, and hMCR-4 (Gantz et al., Biochem. Biophys. Res. Comm. 200:1214-1220 (1994); Gantz et al., J. Biol. Chem. 268:8246-8250 (1993); Gantz et al. J. Biol. Chem. 268:15174-15179 (1993); Haskell-Leuvano et al., Biochem. Biophys. Res. Comm. 204:1137-1142 (1994); each of which is incorporated herein by reference). Vectors for construction of an hMCR-5 expressing cell line are obtained, and a line of HEK 293 cells expressing hMCR-5 is constructed (Gantz, supra, 1994). hMCR-5 has been described previously (Franberg et al., Biochem. Biophys. Res. Commun. 236:489-492 (1997); Chowdhary et al., Cytogenet. Cell Genet. 68:1-2 (1995); Chowdhary et al., Cytogenet. Cell Genet. 68:79-81 (1995), each of which is incorporated herein by reference). HEK 293 cells are maintained in DMEM, 25 mM HEPES, 2 mM glutamine, non-essential amino acids, vitamins, sodium pyruvate, 10% COSMIC CALF SERUM, 100 units/ml penicillin, 100 μg/ml streptomycin and 0.2 mg/ml G418 to maintain selection.

[0180] Before assaying, cells are washed once with phosphate buffered saline (“PBS”; without Ca2+ and Mg2+), and stripped from the flasks using 0.25% trypsin and 0.5 mM EDTA. Cells are suspended in PBS, 10% COSMIC CALF SERUM and 1 mM CaCl2. Cell suspensions are prepared at a density of 2×104 cells/ml for HEK 293 cells expressing hMCR-3, hMCR-4 or hMCR-5, and 1×105 cells/ml for HEK 293 cells expressing hMCR-1. Suspensions are placed in a water bath and allowed to warm to 37° C. for 1 hr.

[0181] Binding assays are performed in a total volume of 250 μl for HEK 293 cells. Control and test compounds are dissolved in distilled water. 125I-HP 467 (50,000 dpm) (2000 Ci/mmol) (custom labeled by Amersham; Arlington Heights Ill.) is prepared in 50 mM Tris, pH 7.4, 2 mg/ml BSA, 10 mM CaCl2, 5 mM MgCl2, 2 mM EDTA and added to each tube. To each tube is added 4×103 HEK 293 cells expressing hMCR-3, hMCR-4 or hMCR-5, or 2×104 cells expressing hMCR-1. Assays are incubated for 2.5 hr at 37° C.

[0182] GF/B filter plates are prepared by soaking for at least one hour in 5 mg/ml BSA and 10 mM CaCl2. Assays are filtered using a Brandel 96-well cell harvester (Brandel Inc.; Gaithersburg, Md.). The filters are washed four times with cold 50 mM Tris, pH 7.4, and the filter plates dehydrated for 2 hr and 35 μl of MICROSCINT is added to each well. Filter plates are counted using a Packard Topcount (Packard Instrument Co.) and data analyzed using GraphPad PRISM v2.0 (GraphPad Software Inc.; San Diego Calif.) and Microsoft EXCEL v5.0a (Microsoft Corp.; Redmond Wash.).

[0183] To assay bicyclic thiophene derivative compounds, binding assays are performed in duplicate in a 96 well format. HP 467 is prepared in 50 mM Tris, pH 7.4, and 121I-HP 467 is diluted to give 100,000 dpm per 50 μl. A bicyclic thiophene derivative compound, is added to the well in 25 μl aliquots. A 25 μl aliquot of 125I-HP 467 is added to each well. A 0.2 ml aliquot of suspended cells is added to each well to give the cell numbers indicate above, and the cells are incubated at 37° C. for 2.5 hr. Cells are harvested on GF/B filter plates as described above and counted.

EXAMPLE 4

Penile Erection due to Administration of a Bicyclic Thiophenederivative Compound

[0184] Adult male rats are housed 2-3 per cage and are acclimated to the standard vivarium light cycle (12 hr. light, 12 hr. dark), rat chow and water for a least a week prior to testing. All experiments are performed between 9 a.m. and noon and rats are placed in cylindrical, clear plexiglass chambers during the 60 minute observation period. Mirrors are positioned below and to the sides of the chambers, to improve viewing.

[0185] Observations begin 10 minutes after an unstraperitoneal injection of either saline or compound. An observer counts the number of grooming motions, stretches, yawns and penile erections (spontaneously occurring, not elicited by genital grooming) and records them every 5 minutes, for a total of 60 minutes. The observer is unaware of the treatment and animals are tested once, with n=6 in each group. Values in the figures represent the group mean and standard error of the mean. HP 228 can be used as a positive control for penile erections. Significant differences between groups are determined by an overall analysis of variance and the Student Neunmann-Keuls post hoc test can be used to identify individual differences between groups (p≦0.05).

[0186] Although the invention has been described with reference to the examples provided above, it should be understood that various modifications can be made by those skilled in the art without departing from the invention. Accordingly, the invention is set out in the following claims.

Claims

1. A single compound of the formula: n is 0 or 1; X is C or N, provided that when X is C, n is 1; and when X is n, n is 0; R1 is selected from the group consisting of C1-C20 alkyl, C2 to C12 alkenyl, C1 to C12 substituted alkyl, C2 to C12 substituted alkenyl, C2 to C12 substituted alkynyl, C3 to C10 cycloalkyl, C3 to C8 substituted cycloalkyl, C7 to C13 cycloalkenyl, C7 to C13 substituted cycloalkanyl, heteroaryl, substituted heteroaryl, C7 to C18 phenylalkyl, C7 to C18 substituted phenylalkyl, phenyl, substituted phenyl, naphthyl, substituted naphthyl, cyclic C2 to C7 alkylene, substituted cyclic C2 to C7 alkylene, cyclic C2 to C7 hetero alkylene, substituted cyclic C2 to C7 heteroalkylene, carboxy and protected carboxy; and, R2 is selected from the group consisting of (a) a carbonyl group coupled with a functional group selected from the group consisting of C1-C20 alkyl, C2 to C12 alkenyl, C1 to C12 substituted alkyl, C2 to C12 substituted alkenyl, C2 to C12 substituted alkynyl, C3 to C10 cycloalkyl, C3 to C8 substituted cycloalkyl, C7 to C13 cycloalkenyl, C7 to C13 substituted cycloalkanyl, heteroaryl, substituted heteroaryl, C7 to C18 phenylalkyl, C7 to C18 substituted phenylalkyl, phenyl, substituted phenyl, naphthyl, substituted naphthyl, cyclic C2 to C7 alkylene, substituted cyclic C2 to C7 alkylene, cyclic C2 to C7 heteroalkylene, substituted cyclic C2 to C7 heteroalkylene, carboxy and protected carboxy; and, (b) phenylsulfonyl, substituted phenylsulfonyl, C1 to C10 alkylsulfonyl, C1 to C10 substituted alkylsulfonyl, C3 to C10 cycloalkylsulfonyl, C2 to C10 substituted alkylsulfonyl; R3 is hydrogen or together with functional Group R1 forms a C2 to C7 substituted or unsubstituted alkylene group; and R4 is hydrogen or together with functional Group R2 forms a C2 to C7 substituted or unsubstituted alkylene group.

2. The compound of claim 1 wherein R1 is selected from the group consisting of 4-butoxybenzyl, benzyl, 2-furyl, 2-naphthyl, 1-adamantanyl, methoxymethyl, benzothiophenyl, phenoxymethyl, 2-chlorobenzyl, 2,3-dimethylbenzyl, 4-cyanobenzyl, cyclobutyl, benzyloxyl, ethyl, thiophene, 3-phenylethyl, 4-ethylbenzyl, 4-tert-butylbenzyl, 3-methoxyphenylmethyl, 3-bromobenzyl, 2,4,5-trifluorobenzyl, 4-n-propylbenzyl, p-toluyl, m-toluyl, o-anisyl, m-anisyl, 3-cyclopentylethyl, propyl, 3-chloropropyl, 4-chlorobutyryl, isopropyl, 2,6-difluorobenzyl, propenyl, 1-ethylpropyl, 3-chloropivalyl, isobutyl, nicotinyl, 2,4,4-trimethylpentanyl, cyclohexanyl, 1-phenylpropyl, cyclopropyl, 2,2-di-n-propylacetyl, butyl, 2,4-difluorobenzyl, 2-ethoxybenzyl, 2-(trifluoromethyl)benzyl, 4-n-hexyloxybenzyl, diphenylmethyl, cyclopentanyl, 1,1-dimethylethyl;

3. The compound of claim 1 wherein R1 is an alkyl group containing 1 to 8 carbon atoms;

4. The compound of claims 1 and 2 wherein R2 is selected from the group consisting of 4-acetamidobenzoyl, 2-(trifluoromethyl)benzoyl, 3,4-difluorobenzoyl, 1-naphthoyl, 3-acetamidobenzoyl, 4-(trifluoromethyl)benzoyl, 2,4-difluorobenzoyl, 7-methoxybenzofuran-2-carbonyl, 2-furoyl, 2-(methylthio)benzoyl, 2-(n-propylthio)nicotinyl, 3-fluoro-2-methyl-benzoyl, 3,4-difluorohydrocinnamyl, 4-chlorocinnamyl, 4-chloro-o-anisyl, 2-chlorocinnamyl, 3-furoyl, 2,5-dimethylphenylacetyl, propionyl, o-toluyl, 2,3-dimethoxybenzoyl, 2-(methylthio)nicotinyl, isobutyryl, crotonyl, thiophene-3-carboxylyl, 2,4-dimethylbenzoyl, cyclohexanecarboxylyl, benzo[b]thiophene-2-carbonyl, isovaleryl, 1-methylindole-3-carbonyl, 2-chlorobenzoyl, 3-chlorobenzoyl, 3-methylthiophene-2-carbonyl, 2,5-dimethoxybenzoyl, 4-chlorobenzoyl, 6-methylnicotinyl, 2-ethoxynicotinyl, cyclobutanecarbonyl, cyclopentylacetyl, 3,5-dimethylisoxazole-4-carbonyl, 1-naphthalenesulfonyl, 2-naphthalenesulfonyl, benzenesulfonyl, 2,5-dichlorobenzenesulfonyl, 2-mesitylenesulfonyl, 4-fluorobenzenesulfonyl, 4-chlorobenzenesulfonyl, 4-methoxybenzenesulfonyl, 4-tert-butylbenzenesulfonyl, p-toluenesulfonyl, methanesulfonyl, beta-styrene sulfonyl, 2,3,5,6-tetramethylbenzenesulfonyl, 3(trifluoromethyl)benzenesulphonyl, 2,5-dimethoxybenzenesulfonyl, o-toluenesulfonyl, p-xylene-2-sulfonyl, 4-ethylbenzenesulfonyl, 4-n-propylbenzenesulfonyl, 4-n-amylbenzenesulfonyl, 4-isopropylbenzenesulphonyl, 2-fluorobenzenesulphonyl, 3-fluorobenzenesulphonyl, 4-chloro-2,5-dimethylbenzenesulphonyl, 2-chlorobenzenesulfonyl, 3-chlorobenzenesulfonyl, m-toluenesulfonyl, 3,4-dimethoxybenzenesulfonyl, 2,3-dichlorobenzenesulfonyl, 2-bromobenzenesulfonyl, 4-(n-butoxy)benzenesulfonyl, 5-chloro-1,3-dimethylpyrazole-4-sulphonyl, 3,5-dimethylisoxazole-4-sulfonyl, 2,4-dichlorobenzenesulfonyl, 5-fluoro-2-methylbenzenesulfonyl, 5-chloro-2-methoxybenzenesulfonyl, 6-methoxy-m-toluenesulfonyl, 4-biphenylsulfonyl, 4-n-butylbenzenesulfonyl, 4-acetylbenzenesulfonyl.

5. A combinatorial library of two or more compounds of the formula:

6. Combinational library of claim 5 wherein R1 is selected from the group consisting of 4-butoxybenzyl, benzyl, 2-furyl, 2-naphthyl, 1-adamantanyl, methoxymethyl, benzothiophenyl, phenoxymethyl, 2-chlorobenzyl, 2,3-dimethylbenzyl, 4-cyanobenzyl, cyclobutyl, benzyloxyl, ethyl, thiophene, 3-phenylethyl, 4-ethylbenzyl, 4-tert-butylbenzyl, 3-methoxyphenylmethyl, 3-bromobenzyl, 2,4,5-trifluorobenzyl, 4-n-propylbenzyl, p-toluyl, m-toluyl, o-anisyl, m-anisyl, 3-cyclopentylethyl, propyl, 3-chloropropyl, 4-chlorobutyryl, isopropyl, 2,6-difluorobenzyl, propenyl, 1-ethylpropyl, 3-chloropivalyl, isobutyl, nicotinyl, 2,4,4-trimethylpentanyl, cyclohexanyl, 1-phenylpropyl, cyclopropyl, 2,2-di-n-propylacetyl, butyl, 2,4-difluorobenzyl, 2-ethoxybenzyl, 2-(trifluoromethyl)benzyl, 4-n-hexyloxybenzyl, diphenylmethyl, cyclopentanyl, and 1,1-dimethylethyl.

7. The combinational library of claim 5 wherein R1 is an alkyl group containing 1 to 8 carbon atoms;

8. The combinational library of claims 5 and 6 wherein R2 is selected from the group consisting of 4-acetamidobenzoyl, 2-(trifluoromethyl)benzoyl, 3,4-difluorobenzoyl, 1-naphthoyl, 3-acetamidobenzoyl, 4-(trifluoromethyl)benzoyl, 2,4-difluorobenzoyl, 7-methoxybenzofuran-2-carbonyl, 2-furoyl, 2-(methylthio)benzoyl, 2-(n-propylthio)nicotinyl, 3-fluoro-2-methyl-benzoyl, 3,4-difluorohydrocinnamyl, 4-chlorocinnamyl, 4-chloro-o-anisyl, 2-chlorocinnamyl, 3-furoyl, 2,5-dimethylphenylacetyl, propionyl, o-toluyl, 2,3-dimethoxybenzoyl, 2-(methylthio)nicotinyl, isobutyryl, crotonyl, thiophene-3-carboxylyl, 2,4-dimethylbenzoyl, cyclohexanecarboxylyl, benzo[b]thiophene-2-carbonyl, isovaleryl, 1-methylindole-3-carbonyl, 2-chlorobenzoyl, 3-chlorobenzoyl, 3-methylthiophene-2-carbonyl, 2,5-dimethoxybenzoyl, 4-chlorobenzoyl, 6-methylnicotinyl, 2-ethoxynicotinyl, cyclobutanecarbonyl, cyclopentylacetyl, 3,5-dimethylisoxazole-4-carbonyl, 1-naphthalenesulfonyl, 2-naphthalenesulfonyl, benzenesulfonyl, 2,5-dichlorobenzenesulfonyl, 2-mesitylenesulfonyl, 4-fluorobenzenesulfonyl, 4-chlorobenzenesulfonyl, 4-methoxybenzenesulfonyl, 4-tert-butylbenzenesulfonyl, p-toluenesulfonyl, methanesulfonyl, beta-styrene sulfonyl, 2,3,5,6-tetramethylbenzenesulfonyl, 3(trifluoromethyl)benzenesulphonyl, 2,5-dimethoxybenzenesulfonyl, o-toluenesulfonyl, p-xylene-2-sulfonyl, 4-ethylbenzenesulfonyl, 4-n-propylbenzenesulfonyl, 4-n-amylbenzenesulfonyl, 4-isopropylbenzenesulphonyl, 2-fluorobenzenesulphonyl, 3-fluorobenzenesulphonyl, 4-chloro-2,5-dimethylbenzenesulphonyl, 2-chlorobenzenesulfonyl, 3-chlorobenzenesulfonyl, m-toluenesulfonyl, 3,4-dimethoxybenzenesulfonyl, 2,3-dichlorobenzenesulfonyl, 2-bromobenzenesulfonyl, 4-(n-butoxy)benzenesulfonyl, 5-chloro-1,3-dimethylpyrazole-4-sulphonyl, 3,5-dimethylisoxazole-4-sulfonyl, 2,4-dichlorobenzenesulfonyl, 5-fluoro-2-methylbenzenesulfonyl, 5-chloro-2-methoxybenzenesulfonyl, 6-methoxy-m-toluenesulfonyl, 4-biphenylsulfonyl, 4-n-butylbenzenesulfonyl, 4-acetylbenzenesulfonyl.

9. A method of preparing a bicyclic thiophene derivative, comprising: (a) acylating hydroxymethyl-resin-bound bicyclic thiophene containing a BOC group with an acylating agent; (b) removing the BOC function group from hydroxymethyl-resin-bound bicyclic thiophene of (a) resulting from (a); (c) acylating the hydroxymethyl-resin-bound bicyclic thiophene of step (b) with a carboxylic acid; (d) cleaving the hydroxymethyl-resin-bound bicyclic thiophene of step (c) using an acid;

10. A method of preparing a bicyclic hydantoin derivative, comprising: (a) acylating hydroxymethyl-resin-bound bicyclic thiophene containing a BOC group with a carbonyl chloride; (b) removing the BOC function group from hydroxymethyl-resin-bound bicyclic thiophene resulting from (a); (c) sultonating the hydroxymethyl-resin-bound bicyclic thiophene of step (b) with a sulfonyl chloride; (d) cleaving the hydroxymethyl-resin-bound bicyclic thiophene of step (c) therein using an acid.