WO2003083063A2 - Analogues de ligands communs: les naphthoates - Google Patents

Analogues de ligands communs: les naphthoates Download PDF

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WO2003083063A2
WO2003083063A2 PCT/US2003/009086 US0309086W WO03083063A2 WO 2003083063 A2 WO2003083063 A2 WO 2003083063A2 US 0309086 W US0309086 W US 0309086W WO 03083063 A2 WO03083063 A2 WO 03083063A2
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compound
nrn
formula
combinatorial library
cooh
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PCT/US2003/009086
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WO2003083063A3 (fr
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Hengyuan Lang
Edcon Chang
Dustin Cefalo
Olga Fryszman
Lin Yu
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Triad Therapeutics, Inc.
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Publication of WO2003083063A3 publication Critical patent/WO2003083063A3/fr

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    • C07C205/57—Compounds containing nitro groups bound to a carbon skeleton the carbon skeleton being further substituted by carboxyl groups having nitro groups and carboxyl groups bound to carbon atoms of six-membered aromatic rings of the carbon skeleton
    • C07C205/59—Compounds containing nitro groups bound to a carbon skeleton the carbon skeleton being further substituted by carboxyl groups having nitro groups and carboxyl groups bound to carbon atoms of six-membered aromatic rings of the carbon skeleton the carbon skeleton being further substituted by singly-bound oxygen atoms
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    • C07C229/00—Compounds containing amino and carboxyl groups bound to the same carbon skeleton
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    • C07C229/06—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton
    • C07C229/18—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton the nitrogen atom of the amino group being further bound to carbon atoms of six-membered aromatic rings
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    • C07C229/56—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to carbon atoms of six-membered aromatic rings of the same carbon skeleton with amino and carboxyl groups bound to carbon atoms of the same non-condensed six-membered aromatic ring with amino and carboxyl groups bound in ortho-position
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    • C07C235/42—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings and singly-bound oxygen atoms bound to the same carbon skeleton
    • C07C235/66—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings and singly-bound oxygen atoms bound to the same carbon skeleton with carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings being part of condensed ring systems and singly-bound oxygen atoms, bound to the same carbon skeleton
    • C—CHEMISTRY; METALLURGY
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    • C07C311/00—Amides of sulfonic acids, i.e. compounds having singly-bound oxygen atoms of sulfo groups replaced by nitrogen atoms, not being part of nitro or nitroso groups
    • C07C311/22—Sulfonamides, the carbon skeleton of the acid part being further substituted by singly-bound oxygen atoms
    • C07C311/29—Sulfonamides, the carbon skeleton of the acid part being further substituted by singly-bound oxygen atoms having the sulfur atom of at least one of the sulfonamide groups bound to a carbon atom of a six-membered aromatic ring
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    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C65/00—Compounds having carboxyl groups bound to carbon atoms of six—membered aromatic rings and containing any of the groups OH, O—metal, —CHO, keto, ether, groups, groups, or groups
    • C07C65/01—Compounds having carboxyl groups bound to carbon atoms of six—membered aromatic rings and containing any of the groups OH, O—metal, —CHO, keto, ether, groups, groups, or groups containing hydroxy or O-metal groups
    • C07C65/105—Compounds having carboxyl groups bound to carbon atoms of six—membered aromatic rings and containing any of the groups OH, O—metal, —CHO, keto, ether, groups, groups, or groups containing hydroxy or O-metal groups polycyclic
    • C07C65/11—Compounds having carboxyl groups bound to carbon atoms of six—membered aromatic rings and containing any of the groups OH, O—metal, —CHO, keto, ether, groups, groups, or groups containing hydroxy or O-metal groups polycyclic with carboxyl groups on a condensed ring system containing two rings
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    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C69/00—Esters of carboxylic acids; Esters of carbonic or haloformic acids
    • C07C69/76—Esters of carboxylic acids having a carboxyl group bound to a carbon atom of a six-membered aromatic ring
    • C07C69/94—Esters of carboxylic acids having a carboxyl group bound to a carbon atom of a six-membered aromatic ring of polycyclic hydroxy carboxylic acids, the hydroxy groups and the carboxyl groups of which are bound to carbon atoms of six-membered aromatic rings
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    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
    • C07D209/04—Indoles; Hydrogenated indoles
    • C07D209/10—Indoles; Hydrogenated indoles with substituted hydrocarbon radicals attached to carbon atoms of the hetero ring
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    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/60—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D213/78—Carbon atoms having three bonds to hetero atoms, with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C07D213/79—Acids; Esters
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    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
    • C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
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    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D249/00—Heterocyclic compounds containing five-membered rings having three nitrogen atoms as the only ring hetero atoms
    • C07D249/02—Heterocyclic compounds containing five-membered rings having three nitrogen atoms as the only ring hetero atoms not condensed with other rings
    • C07D249/08—1,2,4-Triazoles; Hydrogenated 1,2,4-triazoles
    • C07D249/10—1,2,4-Triazoles; Hydrogenated 1,2,4-triazoles with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D249/12—Oxygen or sulfur atoms
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    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D257/00—Heterocyclic compounds containing rings having four nitrogen atoms as the only ring hetero atoms
    • C07D257/02—Heterocyclic compounds containing rings having four nitrogen atoms as the only ring hetero atoms not condensed with other rings
    • C07D257/04—Five-membered rings
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    • C07D309/16—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only ring hetero atom, not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member
    • C07D309/28—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only ring hetero atom, not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D309/30—Oxygen atoms, e.g. delta-lactones
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    • C07D319/00—Heterocyclic compounds containing six-membered rings having two oxygen atoms as the only ring hetero atoms
    • C07D319/04—1,3-Dioxanes; Hydrogenated 1,3-dioxanes
    • C07D319/06—1,3-Dioxanes; Hydrogenated 1,3-dioxanes not condensed with other rings
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B50/00—Methods of creating libraries, e.g. combinatorial synthesis
    • C40B50/14—Solid phase synthesis, i.e. wherein one or more library building blocks are bound to a solid support during library creation; Particular methods of cleavage from the solid support
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/573—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
    • G01N33/5735—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes co-enzymes or co-factors, e.g. NAD, ATP

Definitions

  • the present invention relates generally to receptor/ligand interactions and to combinatorial libraries of ligand compounds.
  • the present invention also relates to the manufacture of naphthoates and combinatorial libraries containing such compounds.
  • Screening for lead compounds involves generating a pool of candidate compounds, often using combinatorial chemistry approaches in which compounds are synthesized by combining chemical groups to generate a large number of diverse candidate compounds that bind to the target or that inhibit binding to the target.
  • the candidate compounds are screened with a drug target of interest to identify lead compounds that bind to the target or inhibit binding to the target.
  • the screening process to identify a lead compound can be laborious and time consuming.
  • Structure-based drug design is an alternative approach to identifying drug candidates.
  • Structure-based drug design uses three-dimensional structural data of the drug target as a template to model compounds that bind to the drug target and alter its activity.
  • the compounds identified as potential drug candidates using structural modeling are used as lead compounds for the development of drug candidates that exhibit a desired activity toward the drug target.
  • Identifying compounds using structure-based drug design can be advantageous when compared to the screening approach in that modifications to the compound can often be predicted by modeling studies.
  • obtaining structures of relevant drug targets and of drug targets complexed with test compounds is extremely time- consuming and laborious, often taking years to accomplish.
  • the long time period required to obtain structural information useful for developing drug candidates is particularly limiting with regard to the growing number of newly discovered genes, which are potential drug targets, identified in genomics studies.
  • the present invention provides compounds that function as mimics to a natural common ligand for a receptor family. These compounds interact with a conserved binding site on multiple receptors within the receptor family.
  • the present invention provides compounds that are common ligand mimics for NAD.
  • NAD is a natural common ligand for many oxidoreductases .
  • compounds of the invention that are common ligand mimics for NAD interact selectively with conserved sites on oxidoreductases .
  • the present invention provides compounds of Formula I,
  • Ri to R 4 each independently are H, alkyl, alkenyl, alkynyl, aryl, heterocycle, COOH, COOAlkyl, CONR 9 R ⁇ 0 , C(0)Rn, OH, OAlkyl, OAc, SH, SRu, S0 3 H, S(0)R u , SO 2 NR 9 R ⁇ 0 , S(0) 2 Rn, NH 2 , NHRn, NR9R10, NHCORu, NR9COR11, N 3 , N0 2 , PH 3 , PH 2 Rn, H 2 P0 4 , H 2 P0 3 , H 2 P0 2 , HP0 4 Rn, P0 2 R ⁇ oRn.
  • R 9 , Rio, and R n each independently are hydrogen, alkyl, alkenyl, alkynyl, aryl, or heterocycle, or R 9 and Rio together with the nitrogen to which they are attached can be joined to form a heterocyclic ring.
  • A is a five or six membered carbocyclic or heterocyclic ring having from 0 to 3 heterocyclic atoms, such as nitrogen, oxygen, or sulfur.
  • the heterocyclic ring can be, for example, any of those described in the definition of "heterocycle” below.
  • the heterocyclic ring can be substituted at one or more positions.
  • the invention provides naphthoate compounds of Formula II,
  • Ri to R 8 each independently are H, alkyl, alkenyl, alkynyl, aryl, heterocycle, COOH, COOAlkyl, CONR 9 R ⁇ 0 ,
  • R 9 , Rio, and Rn each independently are hydrogen, alkyl, alkenyl, alkynyl, aryl, or heterocycle, or R 9 and R ⁇ 0 together with the nitrogen to which they are attached can be joined to form a heterocyclic ring.
  • the present invention provides bi-ligands containing a common ligand mimic and a specificity ligand which interact with distinct sites on a receptor.
  • the present invention provides bi-ligands that are the reaction products of compounds of Formula I with specificity ligands.
  • the invention provides bi-ligands containing the reaction products of compounds of Formula II with specificity ligands.
  • the invention provides methods for preparing bi-ligands that are reaction products of the common ligand mimics of general Formulas I and II and a pyridine dicarboxylate specificity ligand.
  • the present invention provides methods for treating microbial diseases .
  • the present invention further provides combinatorial libraries containing one or more common ligand variants of the compounds of the invention.
  • the combinatorial libraries of the invention contain one or more common ligand variants of the compounds of Formula I.
  • the combinatorial libraries of the invention contain one or more common ligand variants of the compounds of Formula II.
  • the present invention also provides combinatorial libraries comprised of one or more bi- ligands that are reaction products of common ligand mimics and specificity ligands.
  • such combinatorial libraries contain one or more bi-ligands that are the reaction product of compounds of Formula I and specificity ligands.
  • such combinatorial libraries contain one or more bi-ligands that are the reaction product of compounds of Formula II and specificity ligands.
  • the present invention also provides methods for producing and screening combinatorial libraries of bi- ligands for binding to a receptor and families of such receptors.
  • the present invention further provides methods for treating microbial infections or diseases with the compounds of the invention.
  • Figure 1 depicts exemplified common ligand mimics of the present invention.
  • Figure 2 depicts exemplified specificity ligands of the present invention.
  • Figure 3 depicts exemplified bi-ligands of the present invention.
  • Figure 4 shows reaction scheme 1 for the preparation of bi-ligands of the present invention.
  • Figure 5 shows reaction scheme 2 for the preparation of bi-ligands of the present invention.
  • Figure 6 shows reaction scheme 3 for the preparation of bi-ligands of the present invention.
  • Figure 7 shows reaction scheme 4 for the preparation of bi-ligands of the present invention.
  • Figure 8 shows reaction scheme 5 for the preparation of bi-ligands of the present invention.
  • Figure 9 shows reaction scheme 6 for the preparation of bi-ligands of the present invention.
  • Figure 10 shows reaction scheme 7 for the preparation of bi-ligands of the present invention.
  • Figure 11 shows the results of an oxidoreductase assay of selected common ligand mimics of the invention on HMGCoA reductase.
  • Figure 12 shows the results of oxidoreductase assays of selected common ligand mimics of the invention on various oxidoreductases.
  • Figure 13 shows the results of an HMGCoA reductase assay of selected bi-ligands of the invention.
  • Figure 14 shows a reaction scheme for modification of substituents attached to the common ligand mimics of the invention.
  • Figure 15 shows various reaction schemes by which combinatorial libraries of the present invention can be made .
  • Figure 16 shows the binding activities of selected bi-ligand compounds.
  • FIG. 17 shows minimum inhibitory concentration (MIC) assay results for bi-ligand compounds of the invention.
  • Figure 18 shows the MIC activity of a bi-ligand of the invention against E. coli , S . a ureus, and E. fa caelis .
  • the "*" is 20 hour timepoint, assay ends at 24 hours.
  • the present invention is directed to bi-ligands and the development of combinatorial libraries associated with these bi-ligands.
  • the invention can be used advantageously to develop bi-ligands that bind to two distinct sites on a receptor, a common site and a specificity site. Tailoring of the two portions of the bi-ligand provides optimal binding characteristics. These optimal binding characteristics provide increased diversity within a library, while simultaneously focusing the library on a particular receptor family or a particular member of a receptor family.
  • the two portions of the bi-ligand, a common ligand mimic and a specificity ligand act synergistically to provide higher affinity and/or specificity than either ligand alone.
  • the technology of the present invention can be applied across receptor families or can be used to screen for specific members of a family.
  • the present invention can be used to screen libraries for common ligand mimics that bind to any oxidoreductase.
  • the present invention can be used to screen for a particular oxidoreductase that will bind a particular specificity ligand.
  • the present invention provides common ligand mimics that bind selectively to a conserved site on a receptor.
  • the compounds advantageously can be used to develop combinatorial libraries of bi-ligands more efficiently than conventional methods.
  • the present invention takes advantage of NMR spectroscopy to identify the interactions between the common ligand mimic and the receptor, which allows for improved tailoring of the ligand to the receptor.
  • the present invention also provides bi-ligands containing these common ligand mimics.
  • the bi-ligands of the invention contain a common ligand mimic coupled to a specificity ligand. These bi-ligands provide the ability to tailor the affinity and/or specificity of the ligands to the binding sites on the receptor.
  • the present invention further provides combinatorial libraries containing bi-ligands of the invention as well as formation of such libraries from the common ligand mimics of the invention.
  • These libraries provide an enhanced number of bi-ligands that bind multiple members of a receptor family than is provided with standard combinatorial techniques due to specific positioning of the specificity ligand on the common ligand mimic. Optimal positioning of the specificity ligand can be determined through NMR studies of the receptor and the common ligand mimic to be employed.
  • ligand refers to a molecule that can selectively bind to a receptor.
  • selectively means that the binding interaction is detectable over non-specific interactions as measured by a quantifiable assay.
  • a ligand can be essentially any type of molecule such as an amino acid, peptide, polypeptide, nucleic acid, carbohydrate, lipid, or small organic compound.
  • ligand refers both to a molecule capable of binding to a receptor and to a portion of such a molecule, if that portion of a molecule is capable of binding to a receptor.
  • a bi- ligand which contains a common ligand and specificity ligand, is considered a ligand, as would the common ligand and specificity ligand portions since they can bind to a conserved site and specificity site, respectively.
  • ligand excludes a single atom, for example, a metal atom.
  • Derivatives, analogues, and mimetic compounds also are included within the definition of this term.
  • a ligand can be multi-partite, comprising multiple ligands capable of binding to different sites on one or more receptors, such as a bi-ligand.
  • the ligand components of a multi- partite ligand can be joined together directly, for example, through functional groups on the individual ligand components or can be joined together indirectly, for example, through an expansion linker.
  • common ligand refers to a ligand that binds to a conserved site on receptors in a receptor family.
  • a "natural common ligand” refers to a ligand that is found in nature and binds to a common site on receptors in a receptor family.
  • a “common ligand mimic (CLM)” refers to a common ligand that has structural and/or functional similarities to a natural common ligand but is not naturally occurring.
  • a common ligand mimic can be a modified natural common ligand, for example, an analogue or derivative of a natural common ligand.
  • a common ligand mimic also can be a synthetic compound or a portion of a synthetic compound that is structurally similar to a natural common ligand.
  • a "common ligand variant” refers to a derivative of a common ligand.
  • a common ligand variant has structural and/or functional similarities to a parent common ligand.
  • a common ligand variant differs from another variant, including the parent common ligand, by at least one atom. For example, as with NAD and NADH, the reduced and oxidized forms differ by an atom and are therefore considered to be variants of each other.
  • a common ligand variant includes reactive forms of a common ligand mimic, such as an anion or cation of the common ligand mimic.
  • the term "reactive form” refers to a form of a compound that can react with another compound to form a chemical bond, such as an ionic or covalent bond.
  • the common ligand mimic is an acid of the form ROOH or an ester of the form ROOR'
  • the common ligand variant can be ROO " .
  • conserved site on a receptor refers to a site that has structural and/or functional characteristics common to members of a receptor family.
  • a conserved site contains amino acid residues sufficient for activity and/or function of the receptor that are accessible to binding of a natural common ligand.
  • the amino acid residues sufficient for activity and/or function of a receptor that is an enzyme can be amino acid residues in a substrate binding site of the enzyme.
  • the conserved site in an enzyme that binds a cofactor or coenzyme can be amino acid residues that bind the cofactor or coenzyme.
  • receptor refers to a polypeptide that is capable of selectively binding a ligand.
  • the function or activity of a receptor can be enzymatic activity or ligand binding.
  • Receptors can include, for example, enzymes such as kinases, dehydrogenases, oxidoreductases, GTPases, carboxyl transferases, acyl transferases, decarboxylases, transaminases, racemases, methyl transferases, formyl transferases, and ⁇ -ketodecarboxylases .
  • the receptor can be a functional fragment or modified form of the entire polypeptide so long as the receptor exhibits selective binding to a ligand.
  • a functional fragment of a receptor is a fragment exhibiting binding to a common ligand and a specificity ligand.
  • enzyme refers to a molecule that carries out a catalytic reaction by converting a substrate to a product.
  • Enzymes can be classified based on Enzyme Commission (EC) nomenclature recommended by the Nomenclature Committee of the International Union of Biochemistry and Molecular Biology (IUBMB) (see, for example, www.expasy.ch/sprot/enzyme.html) (which is incorporated herein by reference) .
  • EC Enzyme Commission
  • oxidoreductases are classified as oxidoreductases acting on the CH-OH group of donors with NAD + or NADP + as an acceptor (EC 1.1.1); oxidoreductases acting on the aldehyde or oxo group of donors with NAD + or NADP + as an acceptor (EC 1.2.1); oxidoreductases acting on the CH-CH group of donors with NAD + or NADP + as an acceptor (EC 1.3.1); oxidoreductases acting on the CH-NH 2 group of donors with NAD + or NADP + as an acceptor (EC 1.4.1); oxidoreductases acting on the CH-NH group of donors with NAD + or NADP + as an acceptor (EC 1.5.1); oxidoreductases acting on NADH or NADPH (EC 1.6); and oxidoreductases acting on NADH or NADPH with NAD + or NADP + as an acceptor (EC 1.6.1) .
  • Additional oxidoreductases include oxidoreductases acting on a sulfur group of donors with NAD + or NADP + as an acceptor (EC 1.8.1); oxidoreductases acting on diphenols and related substances as donors with NAD + or NADP + as an acceptor (EC 1.10.1); oxidoreductases acting on hydrogen as donor with NAD + or NADP + as an acceptor (EC 1.12.1); oxidoreductases acting on paired donors with incorporation of molecular oxygen with NADH or NADPH as one donor and incorporation of two atoms (EC 1.14.12) and with NADH or NADPH as one donor and incorporation of one atom (EC 1.14.13); oxidoreductases oxidizing metal ions with NAD + or NADP + as an acceptor (EC 1.16.1); oxidoreductases acting on -CH 2 groups with NAD + or NADP + as an acceptor (EC 1.17.1); and oxidoreductases acting on reduced ferredoxin as donor
  • Enzymes can also bind coenzymes or cofactors such as nicotinamide adenine dinucleotide (NAD) and nicotinamide adenine dinucleotide phosphate (NADP) , thiamine pyrophosphate, flavin adenine dinucleotide (FAD) and flavin mononucleotide (FMN) , pyridoxal phosphate, coenzyme A, and tetrahydrofolate or other cofactors or substrates such as ATP, GTP and S-adenosyl methionine (SAM) .
  • enzymes that bind newly identified cofactors or enzymes can also be receptors.
  • receptor family refers to a group of two or more receptors that share a common, recognizable amino acid motif.
  • a motif in a related family of receptors occurs because certain amino acid residues, or residues having similar chemical characteristics, are required for the structure, function and/or activity of the receptor and are, therefore, conserved between members of the receptor family.
  • Methods of identifying related members of a receptor family are well known to those skilled in the art and include sequence alignment algorithms and identification of conserved patterns or motifs in a group of polypeptides, which are described in more detail below.
  • Members of a receptor family also can be identified by determination of binding to a common ligand.
  • the present invention provides bi-ligands that contain a common ligand mimic as described above and a specificity ligand.
  • bi-ligand refers to a ligand comprising two ligands that bind to independent sites on a receptor.
  • One of the ligands of a bi-ligand is a specificity ligand capable of binding to a site that is specific for a given member of a receptor family when joined to a common ligand.
  • the second ligand of a bi- ligand is a common ligand mimic that binds to a conserved site in a receptor family.
  • the common ligand mimic and specificity ligand are bonded together. Bonding of the two ligands can be direct or indirect, such as through a linking molecule or group.
  • a depiction of exemplary bi- ligands is shown in Figure 3.
  • the term “specificity” refers to the ability of a ligand to differentially bind to one receptor over another receptor in the same receptor family.
  • the differential binding of a particular ligand to a receptor is measurably higher than the binding of the ligand to at least one other receptor in the same receptor family.
  • a ligand having specificity for a receptor refers to a ligand exhibiting specific binding that is at least two-fold higher for one receptor over another receptor in the same receptor family.
  • the term “specificity ligand” refers to a ligand that binds to a specificity site on a receptor.
  • a specificity ligand can bind to a specificity site as an isolated molecule or can bind to a specificity site when attached to a common ligand, as in a bi-ligand.
  • the specificity ligand can bind to a specificity site that is proximal to a conserved site on a receptor.
  • the term "specificity site” refers to a site on a receptor that provides the binding site for a ligand exhibiting specificity for a receptor.
  • a specificity site on a receptor imparts molecular properties that distinguish the receptor from other receptors in the same receptor family.
  • the receptor is an enzyme
  • the specificity site can be a substrate binding site that distinguishes two members of a receptor family which exhibit substrate specificity.
  • a substrate specificity site can be exploited as a potential binding site for the identification of a ligand that has specificity for one receptor over another member of the same receptor family.
  • a specificity site is distinct from the common ligand binding site in that the natural common ligand does not bind to the specificity site.
  • linker refers to a chemical group that can be attached to either the common ligand or the specificity ligand of a bi-ligand.
  • the linker provides the functional groups through which the common ligand mimic and specificity ligand are indirectly bound to one another.
  • the linker can be a simple functional group, such as COOH, NH 2 , OH, or the like.
  • the linker can be a complex chemical group containing one or more unsaturation, one or more substituent, and/or one or more heterocyclic atom.
  • Nonlimiting examples of complex linkers are depicted in Tables 6 to 12.
  • the present invention provides common ligand mimics that are common mimics of NAD and combinatorial libraries containing these common ligand mimics-.
  • compounds of the invention are ligands for conserved sites on dehydrogenases and reductases.
  • HMG CoA reductase HMG CoA reductase
  • IMPDH inosine-5'- monophosphate dehydrogenase
  • DOXPR 1-deoxy-D-xylulose- 5-phosphate reductase
  • DHPR dihydrodipicolinate reductase
  • DHFR dihydrofolate reductase
  • IPMDH 3- isopropylmalate
  • GPDH glyceraldehyde-3-phosphate dehydrogenase
  • AR aldose reductase
  • ADH alcohol dehydrogenase
  • LDH lactate dehydrogenase
  • the present invention also provides compounds and combinatorial libraries of compounds of the formula:
  • Ri to R 4 each independently are H, alkyl, alkenyl, alkynyl, aryl, heterocycle, COOH, COOAlkyl, CONR ⁇ 0 Ru, C(0)Ri 2 , OH, OAlkyl, OAc, SH, SR i2 , S0 3 H, S(0)R 12 , SO2NR1 0 R11, S(0) 2 Ri 2 , NH 2 , NHRi 2 , NR 10 Rn, NHCOR ⁇ 2 , NR ⁇ 0 COR ⁇ 2 , N 3 , N0 2 , PH 3 , PH 2 R ⁇ 2 , H 2 P0 4 , H 2 P0 3 , H 2 P0 2 , HP0 4 R i2 , PO2R11R12, CN, or X.
  • R 9 is an oxygen, sulfur, or nitrogen atom, where the nitrogen atom can be substituted, e.g. NR 12 .
  • Rio, R 11 , and R 12 each independently are hydrogen, alkyl, alkenyl, alkynyl, aryl, or heterocycle, or R ⁇ 0 and Rn together with the nitrogen to which they are attached can be joined to form a heterocyclic ring.
  • alkyl means a carbon chain having from one to twenty carbon atoms.
  • R 9 , Rio, and R n each independently can be, for example, hydrogen, alkyl, alkenyl, alkynyl, aryl, or heterocycle, or R 9 and R ⁇ 0 together with the carbon or nitrogen atom to which they are attached can be joined to form a ring.
  • alkyl group present in the compounds of the invention can have one or more of its carbon atoms replaced by a heterocyclic atom, such as an oxygen, nitrogen, or sulfur atom.
  • alkyl as used herein includes groups such as (OCH 2 CH 2 ) n or
  • n has a value such that there are twenty or less carbon atoms in the alkyl group.
  • Similar compounds having alkyl groups containing a nitrogen or sulfur atom are also encompassed by the present invention.
  • alkenyl means an unsaturated alkyl groups as defined above, where the unsaturation is in the form of a double bond.
  • the alkenyl groups of the present invention can have one or more unsaturations .
  • alkynyl means an unsaturated alkyl group as defined above, where the unsaturation is in the form of a triple bond.
  • Alkynyl groups of the present invention can include one or more unsaturations.
  • the compounds of the present invention can include compounds in which Ri to R 8 each independently are complex substituents containing one or more unsaturation, one or more substituent, and/or one or more heterocyclic atom. These complex substituents are also referred to herein as “linkers” or “expansion linkers.” Nonlimiting examples of complex substituents that can be used in the present invention are presented in Tables 6 to 12.
  • aromatic group refers to a group that has a planar ring with 4n+2 pi-electrons, where in is a positive integer.
  • aryl denotes a nonheterocyclic aromatic compound or group. For example, a benzene ring or naphthalene ring.
  • heterocyclic group or “heterocycle” refers to an aromatic compound or group containing one or more heterocyclic atom.
  • Nonlimiting examples of heterocyclic atoms that can be present in the heterocyclic groups of the invention include nitrogen, oxygen and sulfur.
  • heterocydes of the present invention will have from five to seven atoms and can be substituted or unsubstituted.
  • substituents include, for example, those groups provided for Ri to Rs-
  • Nonlimiting examples of heterocyclic groups of the invention include pyroles, pyrazoles, imidazoles, pyridines, pyrimidines, pyridazines, pyrazines, triazines, furans, oxazoles, thiazoles, thiophenes, diazoles, triazoles, tetrazoles, oxadiazoles, thiodiazoles, and fused heterocyclic rings, for example, indoles, benzofurans, benzothoiphenes, benzoimidazoles, benzodiazoles, benzotriazoles, benzotetrazoles, and quinolines .
  • variable "X" indicates a halogen atom.
  • Halogens suitable for use in the present invention include chlorine, fluorine, iodine, and bromine, with bromine being particularly useful.
  • "Ac” denotes an acyl group. Suitable acyl groups can have, for example, an alkyl, alkenyl, alkynyl, aromatic, or heterocyclic group as defined above attached to the carbonyl group.
  • the benzyl ring in Formula I can be substituted with one or multiple substituents. Variation in the substitution on the benzyl ring provides compounds that allow for addition of a specificity ligand to directed sites on the benzyl ring. Direction of the specificity ligand improves the ease and efficiency of manufacture of combinatorial libraries containing bi-ligands having the common ligand mimic bound to a specificity ligand.
  • Ri to R 4 is a substituent other than hydrogen.
  • Ri to R 4 independently can be, alkyl, alkenyl, alkynyl, aryl, heterocycle, COOH, COOAlkyl, CONR 9 R ⁇ 0 , C(0)Rn, OH, OAlkyl, OAc, SH, SRn, S0 3 H, S(0)Rn, SO 2 NR 9 R ⁇ 0 , S(0) 2 Rn, NH 2 , NHRn, NR 9 R ⁇ 0 , NHCORn, NRgCORn, N 3 , N0 2 , PH 3 , PH 2 Ru, H 2 P0 4 , H 2 P0 3 , H 2 P0 2 , HP0 4 Rn, P0 2 R ⁇ oR ⁇ .
  • Ri to R 4 independently can be an amide, a hydroxy group, a thiol group, or an acid group, such as a carboxylic acid.
  • Ri to R 4 independently can be any of the complex substituents provided in Tables 6 to 12.
  • compounds of the invention contain an active hydroxy group, they also can be present in the form of an ether or ester, for example, an alkyl ether or alkyl ester.
  • the invention encompasses compounds in which Ri to R can be an OAlkyl group or a COOAlkyl group.
  • Non-limiting examples of OAlkyl groups include OMe (OCH 3 ) , OEt (OCH 2 CH 3 ), OPr (OCH 2 CH 2 CH 3 ) , and the like.
  • Non-limiting examples of COOAlkyl groups include COOMe, COOEt, COOPr, COOBu, COO-tBu, and the like.
  • two or more of Ri to R 4 are substituents other than hydrogen.
  • the substituent groups can be the same or different.
  • the benzyl ring of the compounds can be substituted with two OAlkyl groups, such as two OMe groups or one OMe group and one OPr group.
  • the benzyl ring of the compounds can be substituted with an OH group and either a COOH or COOAlkyl group. Any combination of the above listed substituents for Ri to R, including complex substituents such as those in Tables 6 to 12, is contemplated by the present invention.
  • the compounds of the invention contain three or more substituents any combination of Ri to R 4 is encompassed by the invention.
  • the invention provides compounds in which Ri to R 4 are not all hydrogen.
  • the invention includes compounds in which at least one of Ri to R 4 is a substituent other than hydrogen.
  • Compounds having complex substituents are encompassed by the invention. The following formulas are representative of such compounds. In each of the formula, any combination of the variables listed can exist. Nonlimiting examples of naphthoate compounds corresponding to formulas la to II are provided in Tables 6 to 12.
  • the invention provides compounds and combinatorial libraries of compounds having formula la
  • D is alkylene, alkenylene, alkynylene, aryl, or heterocycle and Y is OH, NHRn, SRn, COOH, S0 2 OH, X, CN,
  • alkylene alkenylene
  • alkynylene refers to alkyl, alkenyl, and alkynyl groups as defined above in which one additional atom has been removed such that the group is divalent.
  • the invention provides compounds and combinatorial libraries of compounds having formula lb
  • Rio and R n each independently are hydrogen, alkyl, alkenyl, alkynyl, aryl, or heterocycle.
  • the invention provides compounds and combinatorial libraries of compounds having formula Ic
  • Rio and Rn each independently are hydrogen, alkyl, alkenyl, alkynyl, aryl, or heterocycle.
  • the invention provides compounds and combinatorial libraries of compounds having formula Id
  • Rio and Rn each independently are hydrogen, alkyl, alkenyl, alkynyl, aryl, or heterocycle.
  • the invention provides compounds and combinatorial libraries of compounds having formula Ie
  • R 9 , Rio, and Rn each independently are hydrogen, alkyl, alkenyl, alkynyl, aryl, or heterocycle.
  • the invention provides compounds and combinatorial libraries of compounds having formula If
  • Rg, Rio, and Rn each independently are hydrogen, alkyl, alkenyl, alkynyl, aryl, or heterocycle.
  • the invention provides compounds and combinatorial libraries of compounds having formula Ig
  • R ⁇ 0 and Rn each independently are hydrogen, alkyl, alkenyl, alkynyl, aryl, or heterocycle.
  • the invention provides compounds and combinatorial libraries of compounds having formula Ih
  • Rio and Rn each independently are hydrogen, alkyl, alkenyl, alkynyl, aryl, or heterocycle.
  • the invention provides compounds and combinatorial libraries of compounds having formula Ii
  • Rio and Rn each independently are hydrogen, alkyl, alkenyl, alkynyl, aryl, or heterocycle.
  • the invention provides compounds and combinatorial libraries of compounds having formula Ij
  • E is CH 2 CH 2 CH 2 OCH, or CH 2 CH 2 SCH and n is an integer between 1 and 10, inclusive. In certain embodiments of the invention, when n is greater than 4, E is CH 2 CH 2 OCH or CH 2 CH 2 SCH.
  • the invention provides compounds and combinatorial libraries of compounds having formula Ik
  • E is CH 2 , CH 2 CH 2 OCH, or CH 2 CH 2 SCH and n is an integer between 1 and 10, inclusive. In certain embodiments of the invention, when n is greater than 4, E is CH 2 CH 2 OCH or CH 2 CH 2 SCH.
  • the invention provides compounds and combinatorial libraries of compounds having formula II
  • R 6 , R, and Rs each independently are as defined above.
  • the invention provides compounds and combinatorial libraries of compounds having the formula
  • Ri to R 8 each independently are H, alkyl, alkenyl, alkynyl, aryl, heterocycle, COOH, COOAlkyl, CONRgRio, C(0)Ru, OH, OAlkyl, OAc, SH, SRu, S0 3 H, S(0)Ru, SO 2 NR 9 R ⁇ 0 , S(0) 2 Rn, NH 2 , NHRn, NRgRio, NHCORu, NRgCORu, N 3 , N0 2 , PH 3 , PH 2 Ru, H 2 P0 4 , H 2 P0 3 , H 2 P0 2 , HP0 4 Ru, P0 2 R ⁇ oR ⁇ . CN, or X.
  • Rg, Rio, and Rn each independently are hydrogen, alkyl, alkenyl, alkynyl, aryl, or heterocycle, or R 9 and R i0 together with the nitrogen to which they are attached can be joined to form a heterocyclic ring.
  • the compounds of the present invention can include compounds in which R x to R 8 each independently are complex substituents containing one or more unsaturation, one or more substituent, and/or one or more heterocyclic atom. These complex substituents are also referred to herein as "linkers” or "expansion linkers.” Nonlimiting examples of complex substituents that can be used in the present invention are presented in Tables 6 to 12.
  • Ri to R 8 is a substituent other than hydrogen.
  • Ri to R 8 independently can be, alkyl, alkenyl, alkynyl, aryl, heterocycle, COOH, COOAlkyl, CONR 9 R ⁇ 0 , C(0)Rn, OH, OAlkyl, OAc, SH, SRu, S0 3 H, S(0)Ru, SO 2 NR 9 R 10 , S(0) 2 R ⁇ , NH 2 , NHRn, NRgRio, NHCORu, NRgCORu, N 3 , N0 2 , PH 3 , PH 2 Rn, H 2 P0 4 , H 2 P0 3 , H 2 P0 2 , HP0 4 Rn, PO 2 R ⁇ 0 R ⁇ , CN, or X, where R 9 , R ⁇ 0 , and Rn are as defined in Formula I.
  • Ri to R 8 independently can be an amide, a hydroxy group, a thiol group, or an acid group, such as a carboxylic acid.
  • Ri to R 8 independently can be any of the complex substituents provided in Tables 6 to 12.
  • compounds of the invention contain an active hydroxy group, they also can be present in the form of an ether or ester, for example, an alkyl ether or alkyl ester.
  • the invention encompasses compounds in which Ri to R 8 can be an OAlkyl group or a COOAlkyl group.
  • Non-limiting examples of OAlkyl groups include OMe (0CH 3 ) , OEt (OCH 2 CH 3 ), OPr (OCH 2 CH 2 CH 3 ) , and the like.
  • Non-limiting examples of COOAlkyl groups include COOMe, COOEt, COOPr, COOBu, COO-tBu, and the like.
  • two or more of Ri to R 8 are substituents other than hydrogen. In such instances, the substituent groups can be the same or different.
  • the benzyl ring of the compounds can be substituted with two OAlkyl groups, such as two OMe groups or one OMe group and one OPr group.
  • the benzyl ring of the compounds can be substituted with an OH group and either a COOH or COOAlkyl group.
  • Any combination of the above listed substituents for Ri to R 8 including complex substituents such as those in Tables 6 to 12, is contemplated by the present invention.
  • any combination of Ri to R 8 is encompassed by the invention.
  • the invention provides compounds in which Ri to R 8 are not all hydrogen.
  • the invention includes compounds in which at least one of Ri to R 8 is a substituent other than hydrogen.
  • the invention provides compounds containing at least one COOH linker group.
  • D is alkylene, alkenylene, alkynylene, aryl, or heterocycle
  • Rn is hydrogen, alkyl, alkenyl, alkynyl, aryl, or heterocycle.
  • the invention provides compounds and combinatorial libraries of compounds having formula lib
  • Rn is hydrogen, alkyl, alkenyl, alkynyl, aryl, or heterocycle.
  • the invention provides compounds and combinatorial libraries of compounds having formula lie
  • Rio and Rn each independently are hydrogen, alkyl, alkenyl, alkynyl, aryl, or heterocycle.
  • the invention provides compounds and combinatorial libraries of compounds having formula lid
  • Rio and Rn each independently are hydrogen, alkyl, alkenyl, alkynyl, aryl, or heterocycle.
  • the invention provides compounds and combinatorial libraries of compounds having formula lie
  • R, R 9 , Rio, and Rn each independently are hydrogen, alkyl, alkenyl, alkynyl, aryl, or heterocycle.
  • the invention provides compounds and combinatorial libraries of compounds having formula Ilf
  • R, Rg, Rio, and Rn each independently are hydrogen, alkyl, alkenyl, alkynyl, aryl, or heterocycle.
  • the invention provides compounds and combinatorial libraries of compounds having formula Ilg
  • Rio and Rn each independently are hydrogen, alkyl, alkenyl, alkynyl, aryl, or heterocycle.
  • the invention provides compounds and combinatorial libraries of compounds having formula Ilh
  • R ⁇ 0 and Rn each independently are hydrogen, alkyl, alkenyl, alkynyl, aryl, or heterocycle.
  • the invention provides compounds and combinatorial libraries of compounds having formula Hi
  • Rio and Rn each independently are hydrogen, alkyl, alkenyl, alkynyl, aryl, or heterocycle.
  • the invention provides compounds and combinatorial libraries of compounds having formula IIj
  • E is CH 2 , CH 2 CH 2 OCH, or CH 2 CH 2 SCH and n is an integer between 1 and 10, inclusive. In certain embodiments of the invention, when n is greater than 4, E is CH 2 CH 2 OCH or CH 2 CH 2 SCH.
  • the invention provides compounds and combinatorial libraries of compounds having formula Ilk
  • E is CH , CH 2 CH 2 OCH, or CH 2 CH 2 SCH and n is an integer between 1 and 10, inclusive. In certain embodiments of the invention, when n is greater than 4, E is CH 2 CH 2 OCH or CH 2 CH 2 SCH.
  • invention provides compounds and combinatorial libraries of compounds having formula III
  • salt encompasses those salts that form within the carboxylate anions and amine nitrogens and includes salts formed with the organic and inorganic anions and cations discussed below. Furthermore, the term includes salts that form by standard acid-based 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, glutaric, phthalic, tartaric, lauric, stearic, salicyclic, methanesulfonic, benzenesulfonic, sorbic, picric, benzoic, cinnamic, and like acids.
  • organic or inorganic cation refers to counter-ions for the carboxylate anion of a carboxylate salt.
  • the counter-ions are chosen from the sodium, potassium, barium, aluminum, and calcium) ; ammonium and organic cations, such as mono-, di-, and tri-alkyl amines.
  • suitable alkyl amines include, but are not limited to, trimethylamine, cyclohexylamine, dibenzylamine, bis (2-hydroxyethyl) amine, and the like. See for example "Pharmaceutical Salts," Berge et al . , J. Pharm . Sci . , 66:1-19 (1977), which is incorporated herein by reference.
  • 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.
  • any zwitterionic form of the instant compounds formed by a carboxylic acid and an amino group is referred to by this term.
  • a cation for a carboxylate anion will exist when a position is substituted by a (quarternary ammonium) methyl group.
  • the compounds of the invention can also exist as solvates and hydrates. Thus, these compounds can 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.
  • One or more compounds of the invention can be in the biologically active ester form.
  • esters induce increased blood levels and prolong efficacy of the corresponding nonesterified forms of the compounds.
  • the present invention provides bi-ligands that contain a common ligand mimic as described above and a specificity ligand.
  • the common ligand mimic and the specificity ligand can be attached directly or indirectly.
  • the common ligand mimic and specificity ligand are attached via a covalent bond formed from the reaction of one or more functional groups on the common ligand mimic with one or more functional groups on the specificity ligand.
  • Direct attachment of the individual ligands in the bi-ligand can occur through reaction of simple functional groups on the ligands.
  • Indirect attachment of the individual ligands in the bi-ligand can occur through a linker molecule.
  • Such linkers include those provided in Tables 6 to 12.
  • linkers bind to each of the common ligand mimic and the specificity ligand through functional groups on the linker and the individual ligands.
  • Some of the common ligand mimics of the present invention having substituents which include linker molecules, e.g. the common ligand mimics of Tables 6 to 12. Tailoring of the specific type and length of the linker attaching the common ligand mimic and specificity ligand allows tailoring of the bi-ligand to optimize binding of the common ligand mimic to a conservative site on the receptor and binding of the specificity ligand to a specificity site on the receptor.
  • the present invention provides specificity ligands that are specific for NAD receptors and combinatorial libraries containing these specificity ligands.
  • compounds of the invention are ligands for specificity sites on dehydrogenases and reductases like those described above.
  • the present invention provides specificity ligands.
  • specificity ligands may include protecting groups and are reacted with common ligand mimics to form the bi-ligands of the invention. Examples of the specificity ligands useful in the present invention are provided in Figure 2. Specificity ligands, such as those provided in Figure 2 can also exist as salts, or in other reactive forms.
  • Bi-ligands of the invention can be bi-ligands for any receptor.
  • the bi-ligand is a bi-ligand that binds an oxidoreductase.
  • bi-ligands of the present invention comprise a naphthoate compound as a common ligand mimic and a specificity ligand.
  • bi-ligands of the invention can contain a common ligand mimic of Formula I coupled to a specificity ligand, for example those provided in Figure 2.
  • bi-ligands of the invention can contain a common ligand mimic of Formula II coupled to a specificity ligand.
  • the specificity ligand can be any specificity ligand, for example a ligand that binds to a specificity site on an oxidoreductase.
  • the specificity ligand can be a pyridine dicarboxylate . Examples of particular bi- ligands that fall within the invention are provided in Figure 3.
  • Bi-ligands of the present invention can be produced by any feasible method.
  • the compounds of the present invention can be produced by the following methods. These methods are exemplified using a common ligand mimic or Formula I and a pyridine dicarboxylate specificity ligand.
  • a common ligand mimic or Formula I and a pyridine dicarboxylate specificity ligand.
  • variations in such methods can be employed to produce bi-ligands having other common ligand mimics or other specificity ligands .
  • a common ligand mimic of the invention such as a naphthoate compound of Formula I or Formula II can be reacted in the presence of HOBfH 2 0.
  • Suitable solvents include dimethylformamide, tetrahydrofuran, and dichloromethane.
  • the reaction of 4- (2- amino-ethylsulfanyl) -pyridine-2, 6-dicarboxylic acid dimethyl ester can be performed in dimethylformamide with the addition of (HOBfH 2 0) .
  • Triethylamine and 1- dimethylaminopropyl-3-ethyl-carbodiimide (EDCI) are then added to the mixture.
  • the reaction is then stirred at room temperature for a period of about 2 to 40 hours.
  • the reaction can be stirred at room temperature for a period of about 24 hours.
  • reaction precipitate is collected and washed in a mixture of solvent, hydrochloric acid, and methanol. Then, the recovered solid can be suspended in a mixture of alcohol, base, and water, such as a methanol, LiOH, and water mixture. This solution is stirred at room temperature for a period of about 1 to 24 hours until it is homogenous. The solution is then acidified, for example with citric acid or aqueous 2N HCl. The resulting precipitated product can then be filtered, washed with water, and dried.
  • a "combinatorial library” is an intentionally created collection of differing molecules that 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.
  • the present invention provides combinatorial libraries containing two or more compounds.
  • the present invention also provides combinatorial libraries containing three, four, or five or more compounds.
  • the present invention further provides combinatorial libraries that can contain ten or more compounds, for example, fifty or more compounds. If desired, the combinatorial libraries of the invention can contain 100,000 or more, or even 1,000,000 or more, compounds.
  • the present invention provides combinatorial libraries containing common ligand variants of compounds of Formula I.
  • These common ligand variants are active forms of the compounds of Formula I that are capable of binding to a specificity ligand to form a bi-ligand.
  • the common ligand variant can be a compound containing the group COO " .
  • Common ligand variants of the invention include common ligand mimics in which the subsituents on the compounds are complex ligands such as those attached to the compounds listed in Tables 6 to 12.
  • the present invention provides combinatorial libraries containing common ligand variants of compounds of Formula II.
  • These common ligand variants are active forms of the compounds of Formula I that are capable of binding to a specificity ligand to form a bi-ligand.
  • the common ligand variant can be a compound containing the group COO " .
  • Common ligand variants of the invention include common ligand mimics in which the subsituents on the compounds are complex ligands such as those attached to the compounds listed in Tables 6 to 12.
  • the present invention provides combinatorial libraries containing bi- ligands of the invention.
  • the bi-ligands are the reaction product of a common ligand mimic and a specificity ligand which interact with distinct sites on a single receptor.
  • the common ligand mimic can be one or more common ligand mimics for NAD which binds to a conserved site on a dehydrogenase, like ADH.
  • the specificity ligand is one or more ligands which bind a specificity site on ADH.
  • combinatorial libraries can contain bi- ligands having a single common ligand mimic bonded to multiple specificity ligands.
  • the combinatorial libraries can contain bi-ligands having a single specificity ligand bonded to multiple common ligand mimics.
  • the combinatorial libraries can contain multiple common ligand mimics and multiple specificity ligands for one or more receptors.
  • a common ligand mimic of the invention to produce the combinatorial library allows generation of combinatorial libraries having improved affinity and/or specificity. Selection and tailoring of the substituents on the common ligand mimic also allows for production of combinatorial libraries in a more efficient manner than heretofore possible.
  • Bi-ligand libraries of the invention can be prepared in a variety of different ways. For example, two methods employing a resin, such as HOBt resin, carbodiimide resin, or DIEA (diisopropyldiisoamine) resin, can be used to form bi-ligand libraries. In one such method, bi-ligand libraries can be prepared via direct coupling of amines to common ligand mimics of the invention having a carboxylic acid group.
  • a resin such as HOBt resin, carbodiimide resin, or DIEA (diisopropyldiisoamine) resin
  • Bi-ligand libraries can be prepared in the following manner. HOBt resin is swelled in a dry solvent, such as a mixture of dry THF and dry DMF, and added to a solution of a common ligand mimic of the invention that is dissolved in a solvent, such as a mixture of DMF and DIC. The solution is shaken at room temperature overnight and then washed with 3x dry DMF and 3x dry THF. The resin is added to a solution of an amine in a solvent, for example dry DMF. The mixture is shaken again at room temperature overnight. The resin then can be filtered and washed with solvent, and the filtrate can be collected and vacuum dried to provide bi-ligands of the invention.
  • amines useful for the preparation of bi-ligand libraries include those in Table 1.
  • - bi-ligand libraries can be prepared by reacting carboxylic acids to common ligand mimics of the present invention having an amine containing substituent.
  • HOBt resin is swelled a dry solvent, such as dry THF, and added to a solution of a carboxylic acid in a solvent, such as a mixture of dry DMF and DIC. The solution is shaken at room temperature overnight and then washed with 3x dry DMF and lx dry THF. The resin is added to a solution of a common ligand mimic of the invention in a solvent, for example dry DMF. The solution is again shaken at room temperature overnight. The resin then can be filtered and washed with solvent, followed by collection and vacuum drying of the filtrate to provide bi-ligands of the invention.
  • carboxylic acids useful for the preparation of bi-ligand libraries include those in Table 2.
  • bi-ligand libraries of the invention can be built through the direct reaction of isocyanates or thioisocyanates using a combination of solid phase chemistry and solution phase chemistry.
  • Bi-ligand libraries of the invention can further be prepared in the following manner.
  • a solution of an isocyanate or thioisocyanate and a common ligand mimic having an amine of the invention is formed in a solvent, such as DMSO.
  • the isocyanate and common ligand mimic are allowed to react overnight, followed by the addition of aminomethylated polystyrene Resin (NovaBiochem, Cat. No. 01-64-0383) . This mixture is then shaken at room temperature for a period of time, for example about 4 hours.
  • the resin then can be filtered and dried under reduced pressure to yield the desired product.
  • isocyanates and thioisocyanates are provided in Table 3.
  • Bi-ligand libraries of the present invention can be prepared in the following manner.
  • a mixture of DBU, a halopyridine and a thiol is formed in a solvent, such as dioxane.
  • the reaction mixture then is agitated under microwave irradiation at a temperature of 150 to 170°C for a period of about 30 to 40 minutes.
  • the reaction mixture is agitated under microwave irradiation at a temperature of about 170°C for a period of about 40 minutes.
  • the solvent can be removed from the mixture and the resultant oil residue subjected to a column to provide the desired intermediate compound.
  • the intermediate compound then can be suspended in a mixture of water and alcohol, for example a mixture of water and methanol. Lithium hydroxide is added to the solution, which then is refluxed for a period of about 1 to 2 hours, for example a period of about 2 hours.
  • Solvent can be removed from the reaction mixture, and the residue dissolved in water. Dilute hydrochloric acid is added dropwise, forming a white precipitate.
  • the white precipitate is dissolved in a solvent, such as a mixture of dry DMF and DIC.
  • HOBt resin swelled in a solvent, such as dry THF, is then added to the solution, which is shaken at room temperature overnight.
  • the resin then is washed with 3x dry DMF and 2x dry THF and added to a solution of an amine dissolved in a solvent, such as dry DMF.
  • the mixture can be shaken at room temperature overnight, followed by filtration and washing in solvent of the Boc protected intermediate, which then can be collected and vacuum dried.
  • the Boc-protected intermediate is then dissolved in a solvent mixture, for example a mixture of TFA and dichloroethane.
  • a solvent mixture for example a mixture of TFA and dichloroethane.
  • the mixture is then shaken at room temperature for a period of about 15 to 20 minutes, for example a period of about 20 minutes.
  • Solvent can be removed from the mixture to form a deBoc intermediate.
  • HOBt resin swelled in a solvent, such as a mixture of dry THF and dry DMF, is added to a solution of a common ligand mimic of the present invention, dissolved in a solvent, such as a mixture of dry DMF and DIC. This solution then is shaken at room temperature overnight and washed with 3x dry DMF and 3x dry THF.
  • a solvent such as a mixture of dry THF and dry DMF
  • the resin mixture then can be added to a solution of the deBoc intermediate in a solvent, such as dry THF.
  • a solvent such as dry THF.
  • the mixture can be shaken at room temperature overnight, followed by filtration and washing of the resin in a solvent, such as dry DMF.
  • the filtrate then can be collected and vacuum dried to provide bi-ligands of the invention.
  • Nonlimiting examples of amines that are useful in this method include those provided in Table 4.
  • Bi-ligands libraries of the invention can be prepared in the following manner. A mixture of 4- mercaptobenzoic acid and an alkyl bromide is formed in a solvent, such as CH 3 CN. Triethylamine resin (Fluka) then is added to the mixture, which is shaken at room temperature overnight. The resin can be filtered and washed with solvent, followed by collection and vacuum drying.
  • a solvent such as CH 3 CN.
  • the filtrate is dissolved in a solvent, such as a mixture of dry DMF and DIC.
  • HOBt resin swelled in a solvent, such as dry THF, is added to the solution.
  • the solution then is shaken at room temperature overnight and washed with 3x dry DMF and 2x dry THF.
  • the resin then is added to a common ligand mimic of the invention, which has been dissolved in a solvent, such as dry DMF.
  • the solution is shaken at room temperature overnight.
  • the resin then can be filtered and washed with solvent.
  • the filtrate can be collected and vacuum dried to provide bi-ligands of the invention.
  • Nonlimiting examples of alkylhalides useful in this method are provided in Table 5.
  • the present invention is based on the development of bi-ligands that bind to two independent sites on a receptor.
  • the combination of two ligands into a single molecule allows both ligands to simultaneously bind to the receptor and thus can provide synergistically higher affinity than either ligand alone (Dempsey and Snell, Biochemistry 2:1414-1419 (1963); and Radzicka and olfenden, Methods Enzymol. 249:284-303 (1995), each of which is incorporated herein by reference) .
  • the generation of libraries of bi-ligands focused for binding to a receptor family or a particular receptor in a receptor family has been described previously (see WO 99/60404, which is incorporated herein by reference) .
  • the common ligand mimics of the present invention allow for increased diversity of bi-ligand libraries while simultaneously preserving the ability to focus a library for binding to a receptor family.
  • bi-ligands having binding activity for a receptor family it is generally desirable to use a common ligand having relatively modest binding activity, for example, mM to ⁇ M binding activity. This binding activity is increased when combined with a specificity ligand.
  • the common ligand mimic can be modified through the addition of substituents, which can also be called expansion linkers. Substitution of the common ligand mimic allows for tailoring of the bi-ligand by directing the attachment location of the specificity ligand on the common ligand mimic. Tailoring of the bi-ligand in this manner provides optimal binding of the common ligand mimic to the conserved site on the receptor and of the specificity ligand to the specificity site on the same receptor. Through such tailoring, libraries having improved diversity and improved receptor binding can be produced. The bi-ligands contained in such libraries also exhibit improved affinity and/or specificity.
  • a number of formats for generating combinatorial libraries are well known in the art, for example soluble libraries, compounds attached to resin beads, silica chips or other solid supports.
  • the "split resin approach" can be used, as described in U.S. Patent No. 5,010,175 to Rutter and in Gallop et al., J. Med. Chem., 37:1233-1251 (1994), incorporated by reference herein.
  • bi-ligands having diversity at the specificity ligand position have been described previously (see WO 99/60404, WO 00/75364, and US 6,333,149 which issued December 25, 2001).
  • a library of bi-ligands is generated so that the binding affinity of the common ligand mimic and the specificity ligand can synergistically contribute to the binding interactions of the bi-ligand with a receptor having the respective conserved site and specificity site.
  • the bi-ligands are generated with the specificity ligand and common ligand mimic oriented so that they can simultaneously bind to the specificity site and conserved site, respectively, of a receptor.
  • the present invention also provides methods of screening combinatorial libraries of bi-ligands comprising one or more common ligand mimic bound to a variety of specificity ligands and identification of bi- ligands having binding activity for the receptor.
  • the present invention provides methods for generating a library of bi-ligands suitable for screening a particular member of a receptor family as well as other members of a receptor family.
  • BLAST Basic Local Alignment Search Tool
  • ExPASy www.expasy.ch/sprot/prosite.html
  • a third resource for identifying members of a receptor family is Structural Classification of Proteins (SCOP) available at SCOP (scop.mrc- lmb.cam.ac.uk/scop/) (which is incorporated herein by reference) .
  • SCIP Structural Classification of Proteins
  • the next step in development of bi-ligands involves determining whether there is a natural common ligand that binds at least two members of the receptor family, and preferably to several or most members of the receptor family.
  • a natural common ligand for the identified receptor family is already known.
  • dehydrogenases bind to dinucleotides such as NAD or NADP. Therefore, NAD or NADP are natural common ligands to a number of dehydrogenase family members.
  • all kinases bind ATP, and, thus, ATP is a natural common ligand to kinases .
  • At least two receptors in the receptor family are selected as receptors for identifying useful common ligand mimics. Selection criteria depend upon the specific use of the bi-ligands to be produced. Once common ligand mimics are identified, these compounds are screened for binding affinity to the receptor family.
  • Those common ligand mimics having the most desirable binding activity then can be modified by adding substituents that are useful for the attachment and orientation of a specificity ligand.
  • substituents that are useful for the attachment and orientation of a specificity ligand.
  • naphthoates were determined to be common ligand mimics for NAD.
  • These compounds can be modified, for example, by the addition of substituents to the naphthalene ring system.
  • the ring system can be substituted with a COOH group, two OMe groups, or an NHAc group. These groups provide attachment points for the specificity ligand.
  • Substituents added to the ring system can also act as blocking groups to prevent attachment of a specificity ligand at a particular site or can act to orient the specificity ligand in a particular manner to improve binding of the bi-ligand to the receptor.
  • a receptor can be incubated in the presence of a known ligand and one or more potential common ligand mimics.
  • the natural common ligand has an intrinsic property that is useful for detecting whether the natural common ligand is bound.
  • the natural common ligand for dehydrogenases, NAD has intrinsic fluorescence.
  • the known ligand when the natural common ligand does not have an intrinsic property useful for detecting ligand binding, the known ligand can be labeled with a detectable moiety.
  • the natural common ligand for kinases, ATP can be radiolabeled with 32 P, and the displacement of radioactive ATP from an ATP binding receptor in the presence of potential common ligand mimics can be used to detect additional common ligand mimics.
  • Any detectable moiety for example a radioactive or fluorescent label, can be added to the known ligand so long as the labeled known ligand can bind to a receptor having a conserved site.
  • a radioactive or fluorescent moiety can be added to NAD or a derivative thereof to facilitate screening of the NAD common ligand mimics and for bi-ligands of the invention.
  • the library can be screened for binding activity to a receptor in a corresponding receptor family.
  • Methods of screening for binding activity that are well known in the art can be used to test for binding activity.
  • the common ligand mimics and bi-ligands of the present invention can be screened, for example, by the following methods. Screening can be performed through kinetic assays that evaluate the ability of the common ligand mimic or bi-ligand to react with the receptor. For example, where the receptor is and reductase or dehydrogenase for which NAD is a natural common ligand, compounds of the invention can be assayed for their ability to oxidize NADH or NADPH or for their ability to reduce NAD+. Such assays are described more fully in Examples 18 and 19. As disclosed herein, compounds of the invention having binding activity for HMGCoA reductase exhibited antimicrobial activity (see Example 20).
  • Antimicrobial activity can occur through "microbicidal inhibition, " which refers to the ability of a compound to reduce or inhibit the survival of a microorganism by killing or irreversibly damaging it, or through “microbistatic inhibition, " which refers to the ability of a compound to reduce or inhibit the growth or proliferative ability of a target microorganism without necessarily killing it.
  • the invention provides methods of inhibiting microbial growth.
  • the compounds of the invention can be used as antimicrobial agents.
  • the antimicrobial agents can be used to inhibit microbial growth in an environment capable of sustaining survival or growth of a microorganism.
  • An environment capable of sustaining survival or growth of a microorganism means a gaseous, liquid or solid material, including a living organism, in or upon which a microorganism can live or propagate.
  • the types of environments that can be treated using a method of the invention include, for example, a tissue or bodily fluid of an organism such as a human; a liquid such as water or an aqueous solution; a food such as a food crop, a food product or a food extract; and an object such as the surface of an instrument used, for example, to prepare food or to perform surgery; and a gas such as that used for anesthetization in preparation for surgery.
  • a method of the invention encompasses administering to the environment an effective amount of a compound of the invention or analog thereof such that the antimicrobial compound can contact a microorganism in the environment, thereby reducing or inhibiting the ability of the microorganism to grow or survive.
  • a compound of the invention can be used in a variety of procedures for reducing or inhibiting the survival or growth of microorganisms, including the microbicidal inhibition of survival of a microorganism as well as the microbistatic inhibition of growth.
  • a compound of the invention can be used, for example, as a therapeutic agent, a food preservative, a disinfectant or a medicament .
  • an effective amount of a compound of the invention is administered to the environment.
  • the term "effective amount” refers to the amount of a compound that reduces or inhibits the survival or growth of a microorganism in an environment.
  • an effective amount of a compound of the invention produces only minimal effects against the environment, although the level of an acceptable deleterious effect is weighed against the benefit caused by the antimicrobial effect and can be readily determined by one skilled in the art.
  • the invention provides a method of generating a bi-ligand library, comprising coupling a plurality of chemical moieties to a compound of the invention.
  • Methods of generating and coupling a plurality of chemical moities which can be used as putative specifity ligands are well known to those skilled in the art and are exemplified herein.
  • Bi-ligand libraries of the invention can be prepared in a variety of different ways, as described herein above.
  • the compounds of the invention can be used to generate combinatorial libraries, with a plurality of chemical moieties positioned to bind in the specificity site of an enzyme, such as a substrate binding site of an oxidoreductose .
  • a bi-ligand library of the invention can be used in a variety of screening methods, for example, to identify a ligand and/or inhibitor of an NAD cofactor- binding molecule, as well as in methods for optimizing a ligand or inhibitor of an NAD cofactor-binding protein.
  • a variety of experimental approaches can be used for identifying or optimizing a ligand or inhibitor of an NAD cofactor-binding protein.
  • NMR-based methods described in PCT publication WO 03/023392; U.S. Patent Serial No. 6,333,149, and WO 00/75364 are useful for both identifying and optimizing molecules that bind to polypeptides, such as NAD cofactor-binding molecules.
  • the invention provides a method for identifying a ligand that binds to a NAD cofactor binding molecule.
  • the method includes the steps of contacting a NAD cofactor binding molecule with a first compound selected from the compounds of the invention and a specificity ligand under conditions wherein the first compound and the NAD cofactor binding molecule form a bound complex; and detecting magnetization transfer signals between the first compound and the specificity ligand, thereby determining that the first compound and specificity ligand are proximal in a bound complex.
  • Magnetization transfer signals between the molecules can be determined by NMR methods as described, for example, in PCT publication WO 03/023392 (13)A1; U.S. Patent Serial No.
  • These methods can further include obtaining a population of candidate binding compounds by attaching a plurality of chemical moieties, for example, putative specificity ligands to generate bi-ligands, which can be screened for binding activity to various members of a receptor or enzyme family that binds the common ligand of NAD and/or NADP.
  • a plurality of chemical moieties for example, putative specificity ligands to generate bi-ligands, which can be screened for binding activity to various members of a receptor or enzyme family that binds the common ligand of NAD and/or NADP.
  • the invention provides a method of identifying a selective ligand for an NAD cofactor-binding molecule.
  • the method involves contacting an NAD cofactor-binding molecule with a plurality of compounds of a library of the invention, and identifying a compound that binds selectively to the NAD cofactor-binding molecule.
  • the invention provides method for generating a conjugate comprising a compound of the invention coupled to a detectable moiety.
  • the method involves reacting the compound of the invention with a detectable moiety to form a conjugate.
  • a compound of the invention can be coupled to a detectable moiety and used to detect binding to a receptor or enzyme.
  • a compound of the invention that binds to a NAD/NADP cofactor binding site can be coupled to a detectable moiety, and the conjugate containing the compound and the detectable moiety can be used in binding assays to detect binding to an NAD/NADP binding molecule such as an oxidoreductase.
  • a compound of the invention coupled to a detectable moiety can be used screen candidate common ligands by competitive binding assays.
  • a "detectable moiety” refers to a molecule that can be detected through physical or chemical means. Any detectable moiety can be employed in the present invention, so long as the detectable moiety does not interfere with the binding of a ligand to its binding partner, for example, a receptor or enzyme that binds an NAD or NADP cofactor.
  • the detectable moiety can be a molecule detectable by analytical methods including, for example, fluorescent tags; fluorescent proteins, such as green fluorescent protein; radioactive tags; ferromagnetic substances; luminescent and chemiluminescent tags, chromophores and colorimetric indicators; detectable binding agents, such as members of a binding pair like biotin/streptavidin or antibodies/antigens. Methods of coupling or conjugating a detectable moiety to a compound of the invention are well know to those skilled in the art (see, for example, Hermanson, Bioconjugate Techniques, Academic Press, San Diego (1996) ) .
  • detectable moiety useful in the present invention is a fluorescent tag.
  • Fluorescent tags are well known in the art and are described, for example, in Hermanson, Bioconjugate Techniques, pp. 297- 364, Academic Press, San Diego (1996) .
  • Fluorescent molecules useful in the invention include, but are not limited to, fluorescein and fluorescein derivatives; rhodamine and rhodamine derivatives; coumarin and coumarin derivatives; BODIPYTM (4, -difluoro-4-bora-3a, 4a- diaza-s-indacene) and BODIPYTM derivatives (Molecular Probes; Eugene, OR) ; Cascade BlueTM (8-methoxypyrene- 1, 3, 6-trisulfonic acid) and derivatives thereof (Molecular Probes); Lucifer Yellow (3, 6-disulfonate-4- amino-napthalimide) and derivatives thereof; Alexa fluor dyes (Molecular Probes) and CyDye fluorescent dyes
  • fluorescent moieties include fluorescein isothiocyanate (FITC) , Lissamine, Rhodamine B, Oregon Green®, Rhodamine GreenTM, Rhodamine Red-X, Texas Red and related compounds, tetramethylrhodamine, and the like.
  • FITC fluorescein isothiocyanate
  • detectable moiety useful in the present invention is a fluorescent protein.
  • Fluorescent proteins include, but are not limited to, green fluorescent protein and derivatives thereof as well as phycobiliproteins, and derivatives thereof, such as phycoerythrin and phycocyanin.
  • Nonlimiting examples of chromophores suitable as detectable moieties include phenolphthalein, malachite green, phytochromes and apophytochromes, yellow protein chromophore, melanophores, and phenanthrolines .
  • Another detectable moiety is a dye that can be visualized. Exemplary dyes include eosins, erythrosins, and the like.
  • colorimetric indicators such as chemiluminescent molecules, which are useful as detectable moieties include, but are not limited to, 1, 2-dioxetane and luminol.
  • radioactive tags suitable as detectable moieties include 32 P, 33 P, 35 S, 3 H, 14 C, 125 I, 59 Fe, and 18 F.
  • the invention provides a method of identifying an inhibitor of NAD cofactor binnding to an NAD cofactor-binding molecule.
  • the method involves contacting an NAD cofactor-binding molecule with a candidate inhibitor and a conjugate comprising a compound of the invention coupled to a detectable moiety; measuring the conjugate in the presence of the NAD cofactor-binding molecule and the absence of the candidate inhibitor; comparing the measured conjugate in the presence of the NAD cofactor-binding molecule and the candidate inhibitor to the conjugate in the presence of the NAD cofactor-binding molecule and the absence of the candidate inhibitor, wherein a decrease in the measured conjugate indicates binding of the candidate inhibitor to the NAD cofactor-binding molecule.
  • a variety of approaches can be used to perform a competive binding assay using a conjugate coupled to a compound of the invention coupled to a detectable moiety. Such approaches can involve detecting a conjugate coupled to a compound of the invention in solution or solid-state forms and using a variety of different detection methods, depending on the particular detectable moiety selected. Those skilled in the art will be able to determine an appropriate detection method for a selected detectable moiety. It is understood that a candidate inhibitor can comprise a compound of the invention and can be screened as described above. An identified compound that has competitive binding activity can be further screened for enzyme inhibitory activity using methods well known to those skilled in the art.
  • a method for treating a patient having an NAD cofactor-binding molecule-associated condition involves administering to the patient an effective amount of a compound of the invention, wherein symptoms or progression of the NAD cofactor-binding molecule- associated condition are reduced.
  • an exemplary NAD cofactor-binding molecule-associated condition is bacterial infection.
  • bi-ligands containing compounds of the invention can be used as antimicrobial agents to inhibit the growth of a microorganism, as disclosed herein.
  • Butoxycarbonylmethyl-2, 2-dimethyl- [1,3] dioxan- 4-ylmethanol (compound 1, 1.38 g, 5.3 mmol) was dissolved in CH 2 C1 2 (25 ml) under N 2 and chilled to a temperature of 0°C.
  • Triethylamine (883 ⁇ l, 6.3 mmol) was added to the solution, followed by the addition of methanesulfonyl chloride (450 ⁇ l, 5.8 mmol). The solution was allowed to warm to room temperature while stirring overnight and then was quenched with water.
  • the mixture was diluted with methylene chloride, and the organic layer was washed with water and brine, dried (Na 2 S0 4 ) , and concentrated.
  • the crude product was redissolved in tetrahydrofuran (THF) , and tetrabutylammonium azide was added (2.26 g, 7.95 mmol). The mixture was heated to a temperature of 50 °C for a period of three days, followed by dilution with diethyl ether and water. The organic layers were washed with water and brine and then dried.
  • the crude product was purified by flash chromatography on silica gel eluted with -85:15 hexanes/ethyl acetate to provide 985 mg (65%) of the product azide.
  • Step b Formation of (6-aminomethyl-2 , 2-dimethyl- [l,3]dioxan-4-yl) -acetic acid tert-butyl ester
  • the solution was allowed to warm to room temperature with stirring overnight, then quenched with water.
  • the mixture was diluted with methylene chloride, and the organic layer was washed with water and brine, dried (Na 2 S0 4 ) , and concentrated.
  • the crude product was redissolved in N,N-dimethylformamide (DMF), and sodium azide was added (1.42 g, 21.9 mmol).
  • the mixture was stirred overnight and then diluted with a mixture of methylene chloride and water.
  • the organic layer was washed with water three times and brine once, and dried.
  • Step B Formation of C- [3- (4-fluoro-phenyl) -1-isopropyl- lH-indol-2-yl] -methylamine
  • the azide (70 mg, 0.23 mmol) was dissolved in THF (30 ml) and treated with diphenylphosphinopolystyrene (250 mg, 0.25 mmol) and water (1 drop, catalytic) overnight.
  • the resin was filtered and rinsed with diethyl ether and methylene chloride. The filtrate was dried (Na 2 S0 4 ) , and concentrated.
  • R, Ri, and R 2 hydrogen, alkyl, alkenyl, alkynyl, aryl, and heterocyclic
  • R in the compounds is alky, alkenyl, alkynyl, aromatic, or heterocyclic.
  • the variables E, F, Y, and n can have the values provided in Table 8 above.
  • the variables E, F, Y, and n can have the values provided in Table 8 above.
  • the variables E, F, Y, and n can have the values provided in Table 8 above.
  • This example provides a general procedure for preparing bi-ligand libraries from common ligand mimics of the invention according to the reaction scheme presented in Figure 15a.
  • Compound numbers correspond to the numbers in the figure.
  • HOBt resin is in cry DMF.
  • the resin then is added to a solution of compound 10 dissolved in a mixture of dry DMF and DIC (N, ' -diisopropylcarbodii ide) .
  • the solution is shaken at room temperature for a period of about 2 to 20 hours and then washed three times with dry DMF and three times with dry THF.
  • HOBt resin is swelled in dry DMF.
  • the resin is added to a solution of carboxylic acid (1-naphthalene acetic acid) dissolved in a mixture of dry DMF and DIC.
  • the solution is shaken at room temperature overnight and washed with 3x dry DMF and lx dry THF.
  • the naphthoate compounds depicted in Figure 13 were prepared in accordance with the methods of Examples 1 and 5. The compounds were screened for binding to the following enzymes: dihydrodipicolinate reductase (DHPR) , alcohol dehydrogenase (ADH) , dihydrofolate reductase (DHFR), l-deoxy-D-xylulose-5-phosphate reductase (DOXPR), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) , inosine-5' -monophosphate dehydrogenase (IMPDH) , and HMG CoA reductase (HMGCoAR) .
  • DHPR dihydrodipicolinate reductase
  • ADH alcohol dehydrogenase
  • DHFR dihydrofolate reductase
  • DOXPR l-deoxy-D-xylulose-5-phosphate reductase
  • GPDH
  • the compounds were screened using a kinetic protocol that spectrophotometrically evaluates oxidation of NADPH.
  • DHPR dihydrodipicolinate synthase
  • the L-ASA (L-aspartate semialdehyde) solution was prepared in the following manner. 180 ⁇ M stock solution of ASA was prepared. 100 ⁇ l of the ASA stock solution was mixed with 150 ⁇ l of concentrated NaHC0 3 and 375 ⁇ l of H 2 0. For use in the assay, 28.8 mM L-ASA was equal to 625 ⁇ l of the solution. The L-ASA stock solution was kept at a temperature of -20°C. After dilution, the pH of the 28.8 mM solution was checked and maintained between 1 and 2.
  • the DHPS reaction was monitored at 340 nm prior to and after addition of the inhibitor to detect background reaction with the inhibitor.
  • the solution for background detection was a 945 ⁇ l solution containing 0.1 HEPES (pH 7.8), 1 mM pyruvate, 6 ⁇ M NADPH, 40 ⁇ M L-ASA, and 7 ⁇ l of 1 mg/ml DHPS at 25°C in the volumes provided above.
  • the sample solution was then mixed and incubated for 10 minutes.
  • 500 nM solutions of the inhibitors and enough DMSO to provide a final DMSO concentration of 5% of the total assay volume were added.
  • the solution was mixed and incubated for an additional 6 minutes.
  • DHPR samples 5 ⁇ l of the diluted DHPR enzyme were added. The sample was mixed for 20 seconds and then the reaction was run for 10 minutes. After a 50 second lag, the samples were read in a Cary spectrophotometer at 340 nm. Reading of the samples was continued until 300 seconds.
  • Cuvette #1 contained the control reaction (no inhibitor)
  • cuvette #2 contained the positive control reaction in which Cibacron Blue at 2.58 ⁇ M was substituted for inhibitor to yield 70 to 80% inhibition.
  • the substrate was kept at a level at least 10 times the Km. The final concentration of L-ASA was about 1 mM.
  • the compounds were screened using a kinetic protocol that spectrophotometrically evaluates reduction of NAD+.
  • the ADH reaction was monitored at 340 nm prior to and after addition of the inhibitor to detect background reaction with the inhibitor.
  • Solutions of 100 ⁇ l of the inhibitors in DMSO were prepared to provide a final DMSO concentration of 5% of the total assay volume. These solutions were incubated for 6 minutes at 25°C in a 990 ⁇ l of a solution containing 0.1 M HEPES, pH 8.0, 80 ⁇ M NAD+, and 130 mM of ethanol.
  • the reaction was then initiated with 10 ⁇ l of ADH from Bakers Yeast (3.3 ⁇ g/ml; 1:400 dilution of 1.0 mg/ml). After the enzyme was added, the solution was mixed for 20 seconds, and the reaction was run for 10 minutes.
  • the samples were read in a Cary spectrophotometer at 340 nm. Reading of the samples was continued until 300 seconds.
  • Cuvette #1 contained the control reaction (no inhibitor)
  • cuvette #2 contained the positive control reaction in which Cibacron Blue at 15.5 ⁇ M was substituted for inhibitor to yield 50 to 60% inhibition.
  • the substrate was kept at a level at least 10 times the Km. The final concentration of pyruvate was about 2.5 mM. Where only a simple read was desired, as in the case of NAD+ concentration determination, 13 ⁇ l (10 M stock) of ethanol was used to drive the reaction, and 10 ⁇ l of pure enzyme (1 mg/ml) was used.
  • NAD+ was soluble at 2 mM, which allowed the concentration determination step to be skipped.
  • the procedure was as follows. All of the ingredients except for the enzyme were mixed together. The solution was mixed well and the absorbance at 340 nm read. The enzyme was added and read again at OD 340 after the absorbance stopped changing, generally 10 to 15 minutes after the enzyme was added.
  • the compounds were screened using a kinetic protocol that spectrophotometrically evaluates oxidation of NADH.
  • the DHFR reaction was monitored at 340 nm prior to and after addition of the inhibitor to detect background reaction with the inhibitor.
  • Solutions of 100 ⁇ l of the inhibitors in DMSO were prepared to provide a final DMSO concentration of 5% of the total assay volume. These solutions were incubated for 6 minutes at 25°C in a 992 ⁇ l of a solution containing 0.1 M Tris-HCl, pH 7.0, 150 mM KCI, 5 ⁇ M H 2 folate, and 52 ⁇ M NADH.
  • the oxidation reaction was then initiated with 8 ⁇ l of DHFR (0.047 mg/ml) . After the enzyme was added, the solution was mixed for 20 seconds, and the reaction was run for 10 minutes.
  • the compounds were screened using a kinetic protocol that spectrophotometrically evaluates oxidation of NADPH.
  • Inhibitor 15 m-M 100 ⁇ M 6.7 ⁇ l (0.67% DMSO
  • the DOXPR reaction was monitored at 340 nm prior to and after addition of the inhibitor to detect background reaction with the inhibitor.
  • Solutions of the inhibitors in DMSO were prepared to provide a final DMSO concentration of 5% of the total assay volume. These solutions were incubated for 6 minutes at 25°C in a 990 ⁇ l of a solution containing 0.1 M HEPES, pH 7.4, 1 mM MnCl 2 1.15 mM DOXP, and 8 ⁇ M NADPH.
  • the oxidation reaction was then initiated with 10 ⁇ l of DOXP reductoisomerase (10 ⁇ g/ml) . After the enzyme was added, the solution was mixed for 20 seconds, and the reaction was run for 10 minutes.
  • the samples were read in a Cary spectrophotometer at 340 nm. Reading of the samples was continued until 300 seconds.
  • Cuvette #1 contained the control reaction (no inhibitor)
  • cuvette #2 contained the positive control reaction in which Cibacron Blue at 10.32 ⁇ M was substituted for inhibitor to yield 70 to 80% inhibition.
  • the substrate was kept at a level at least 10 times the Km.
  • the compounds were screened using a kinetic protocol that spectrophotometrically evaluates reduction of NAD+.
  • Stock solutions of each of the reagents were prepared in the following concentrations. Dilutions of the stock solutions were prepared prior to running the assay in the concentrations indicated below.
  • the GAPDH reaction was monitored at 340 nm prior to and after addition of the inhibitor to detect background reaction with the inhibitor.
  • Solutions of 100 ⁇ l of the inhibitors incubated for 6 minutes at 25°C in a 990 ⁇ l of a solution containing 125 mM triethanolamine, pH 7.5, 145 ⁇ M glyceraldehyde 3-phosphate (GAP), 0.211 mM NAD, 5 mM sodium arsenate, and 3mM ⁇ -metcaptoethanol (2- BME) .
  • GAP glyceraldehyde 3-phosphate
  • the reaction was then initiated with 10 ⁇ l of E. coli GAPDH (1:200 dilution of 1.0 mg/ml). After the enzyme was added, the solution was mixed for 20 seconds, and the reaction was run for 10 minutes.
  • the tube was then immersed, while shaking, in the boiling water for 3 minutes. Next, the tube was placed in an ice bath to cool for 5 minutes. As the sample cooled, a resin settled to the bottom of the test tube, allowing removal of the supernatant with a pasteur pipette. The supernatant was filtered through a 0.45 or 0.2 ⁇ M cellulose acetate syringe filter.
  • the filtered supernatant was retained, and another 1 ml of dH 2 0 was added to the resin tube. The tube was then shaken and centrifuged for 5 minutes at 3,000 rpm. The supernatant was again removed with a pasteur pipette and passed through a 0.45 or 0.2 ⁇ M cellulose acetate syringe filter. The two supernatant aliquots were then pooled to provide a total GAP concentration of about 50 mM. The GAP was then divided into 100 ⁇ l aliquots and stored at -20°C until use.
  • the compounds were screened using a kinetic protocol that spectrophotometrically evaluates reduction of NAD+.
  • Stock solutions of each of the reagents were prepared in the following concentrations. Dilutions of the stock solutions were prepared prior to running the assay in the concentrations indicated below.
  • the IMPDH reaction was monitored at 340 nm prior to and after addition of. the inhibitor to detect background reaction with the inhibitor.
  • Solutions of 100 ⁇ l of the inhibitors in DMSO were prepared to provide a final DMSO concentration of 5% of the total assay volume. These solutions were incubated for 6 minutes at 37°C in a 992 ⁇ l of a solution containing 0.1 M Tris-HCl, pH 8.0, 0.25 M KCI, 0.3% glycerol, 30 ⁇ M NAD+, and 600 ⁇ M IMP (inosine monophosphate) .
  • the reaction was then initiated with 8 ⁇ l of IMPDH (0.75 ⁇ g/ml). After the enzyme was added, the solution was mixed for 20 seconds, and the reaction was run for 10 minutes.
  • the samples were read in a Cary spectrophotometer at 340 nm. Reading of the samples was continued until 300 seconds.
  • Cuvette #1 contained the control reaction (no inhibitor)
  • cuvette #2 contained the positive control reaction in which Cibacron Blue was substituted for inhibitor.
  • the substrate was kept at a level at least 10 times the Km.
  • the compounds were screened using a kinetic protocol that spectrophotometrically evaluates oxidation of NADPH.
  • the HMGCoAR reaction was monitored at 340 nm prior to and after addition of the inhibitor to detect background reaction with the inhibitor.
  • Solutions of 500 nM of the inhibitors in DMSO were prepared to provide a final DMSO concentration of 2% of the total assay volume. These solutions were incubated for 6 minutes at 25°C in a 994 ⁇ l of a solution containing 25 mM KH 2 P0, pH 7.5, 160 ⁇ M HMGCoA, 13 ⁇ M NADPH, 50 m-M NaCl, 1 mM EDTA, and 5mM DTT. The reaction was then initiated with 5 ⁇ l of
  • HMGCoAR enzyme (1:40 dilution of 0.65 mg/ml). After the enzyme was added, the solution was mixed for 20 seconds, and the reaction was run for 10 minutes. After a 50 second lag, the samples were read in a Cary spectrophotometer at 340 nm. Reading of the samples was continued until 300 seconds.
  • Cuvette #1 contained the control reaction (no inhibitor)
  • cuvette #2 contained the positive control reaction in which Cibacron Blue at 2.05 ⁇ M was substituted for inhibitor to yield 50 to 70% inhibition.
  • the substrate was kept at a level at least 10 times the Km.
  • IC 50 data for various compounds against various oxidoreductases are presented in Figure 12.
  • the compounds shown in Figure 12 exhibited binding activity for various oxidoreductases.
  • Several of the compounds exhibited specificity for HMGCoA reductase of greater than 2-fold relative to the other oxidoreductases tested. These results show that several compounds have specific binding activity for HMGCoA reductase.
  • This example describes the screening of naphthoate common ligand mimics for binding activity to a variety of oxidoreductases.
  • ICso data for these compounds are presented in Figure 13.
  • the compounds shown in Figure 13 exhibited binding activity for HMGCoA reductase.
  • One compound exhibited and IC50 of less than 600 nM. Therefore, the compounds exhibited high affinity binding for HMGCoA reductase.
  • the antimicrobial susceptibilities of Escherichia coli, Staphylococcus aureus, and Enterococcus faecalis were determined using the microdilution method set out by the National Committee for Clinical Laboratory Standards (NCCLS) .
  • NCCLS National Committee for Clinical Laboratory Standards
  • the bacterial strains tested were the ATCC strains used by NCCLS as clinical quality control (QC) standards.
  • the bi-ligand inhitors were resuspended in dimethylsulfoxide (DMSO) at a final concentration of 2 mg/ml.
  • DMSO dimethylsulfoxide
  • Bacteria were grown to mid-log phase and diluted to ⁇ 1 X 10 6 CFU/ml in fresh Mueller- Hinton broth (MHB) .
  • the inhibitors were diluted into sterile MHB at a concentration of 120 ⁇ g/ml; a 2-fold dilution series was prepared.
  • Microtiter plate samples consisted of equal volumes of the bacterial and drug suspensions .
  • Samples were run in triplicate with drug concentration ranging from 60-0.94 ⁇ g/ml.
  • the blank consisted of MHB plus drug candidate.
  • Two additional controls were run in triplicate.
  • a gentamicin MIC was run as an internal plate control to ensure the QC strains were inhibited by a known drug in the correct MIC range.
  • An MHB +DMSO dilution series was run to ensure that the growth inhibition was not caused by the inclusion of DMSO in the dilution series.
  • Shown in Figure 16 are the respective binding activities of selected bi-ligand compounds.
  • the IC50 binding activity for HMGCoA reductase are shown.
  • HMGCoA reductase was selected as a target enzyme for testing antimicrobial activity of bi-ligand compounds because HMGCoA reductase is known to be expressed in Staphylococcus and Streptococcus as well as Enterococcus . Therefore, compounds which bind to and/or inhibit HMGCoA reductase are candidate compounds that can be tested for antimicrobial activity.
  • Such compounds can be particularly useful for treating antibiotic resistant organisms such as methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus .
  • MIC Minimum inhibitory concentration assays were performed as described above. Compounds were tested against three organisms: Escherichia coli (HMGCoA reductase negative) , Staphylococcus aureus (HMGCoA reductase positive) , and Enterococcus faecalis (HMGCoA reductase positive) . A 2-fold dilution series was performed as described above, starting at 60 ⁇ g/ml (3% DMSO) , for each compound. Bacteria were tested at each concentration in triplicate using and endpoint assay.
  • Figure 18 shows the activity another compound tested for MIC activity against E. coli , S . aureus, and E.rioselis .
  • the compound inhibited Enterococcus and Staphylococcus but not E. coli .

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Abstract

L'invention concerne des analogues de ligands communs agissant en tant que ligands communs pour une famille de récepteurs. L'invention concerne également des bi-lingands contenant ces analogues de ligands communs. Les bi-ligands de l'invention présentent une affinité accrue et/ou une sélectivité de liaison de ligand par rapport à un récepteur ou à une famille de récepteurs, par le biais de l'action synergique de l'analogue de ligand commun et du ligand de spécificité qui compose ce bi-ligand. L'invention concerne des bibliothèques combinatoires contenant les analogues de ligands communs et les bi-ligands de l'invention. En outre, l'invention concerne encore des procédés de fabrication des analogues de ligands communs et des bi-ligands de l'invention, et des méthodes d'essai des bibliothèques combinatoires de l'invention.
PCT/US2003/009086 2002-03-22 2003-03-21 Analogues de ligands communs: les naphthoates WO2003083063A2 (fr)

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006117657A1 (fr) * 2005-05-03 2006-11-09 Ranbaxy Laboratories Limited Derives de triazolone utilises comme agents anti-inflammatoires

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2848296A (en) * 1955-11-17 1958-08-19 Goodrich Co B F Process for the basic dyeing of vinylidene cyanide interpolymers
US3285706A (en) * 1960-09-26 1966-11-15 Alliance Color And Chemical Co Continuous fusion apparatus
US4002675A (en) * 1974-02-15 1977-01-11 Basf Aktiengesellschaft Manufacture of 2-hydroxy-naphthalene-3-carboxylic acid
US5344843A (en) * 1991-05-09 1994-09-06 Hoffman-La Roche Inc. Cat-1 inhibitors, pharmaceutical compositions and methods of use
US6297348B1 (en) * 1999-08-30 2001-10-02 Shell Oil Company Closely linking a NDA process with a pen process

Family Cites Families (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE838290C (de) * 1950-05-28 1952-05-08 Naphtol Chemie Offenbach Verfahren zur Herstellung von Arylamiden der 2-Oxynaphthalin-1-carbonsaeure
AT368678B (de) * 1980-08-05 1982-10-25 Nitrokemia Ipartelepek Antidotum zum schutz von kulturpflanzen gegen die schaedigende wirkung von herbiziden
RO93222B1 (ro) * 1986-03-22 1987-12-31 COMBINATUL DE îNGRASAMINTE CHIMICE Procedeu pentru obtinerea acidului 1, 4-dihidroxi-2-naftoic
EP0478197A1 (fr) * 1990-09-17 1992-04-01 MITSUI TOATSU CHEMICALS, Inc. Procédé pour contrôler le rapport des isomères d'acides carboxyliques hydroxyaromatiques
JPH05148222A (ja) * 1991-11-29 1993-06-15 Fujirebio Inc ナフトエ酸誘導体
JPH1014589A (ja) * 1996-07-08 1998-01-20 Unitika Ltd キャンディダ・リポリティカｈｎａ−１１０株及び２，６−ジヒドロキシ−３−ナフトエ酸の製造方法
JP4105305B2 (ja) * 1997-10-03 2008-06-25 上野製薬株式会社 芳香族ヒドロキシカルボン酸の製造方法
JP3824288B2 (ja) * 1998-05-26 2006-09-20 富士写真フイルム株式会社 ポジ型感光性樹脂組成物

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2848296A (en) * 1955-11-17 1958-08-19 Goodrich Co B F Process for the basic dyeing of vinylidene cyanide interpolymers
US3285706A (en) * 1960-09-26 1966-11-15 Alliance Color And Chemical Co Continuous fusion apparatus
US4002675A (en) * 1974-02-15 1977-01-11 Basf Aktiengesellschaft Manufacture of 2-hydroxy-naphthalene-3-carboxylic acid
US5344843A (en) * 1991-05-09 1994-09-06 Hoffman-La Roche Inc. Cat-1 inhibitors, pharmaceutical compositions and methods of use
US6297348B1 (en) * 1999-08-30 2001-10-02 Shell Oil Company Closely linking a NDA process with a pen process

Non-Patent Citations (17)

* Cited by examiner, † Cited by third party
Title
DATABASE CAPLUS [Online] BALLINI ET AL.: 'Zeolite HSZ-360 as a new reusable catalyst for the direct acetylation of alcohols and phenols under solventless conditions', XP002972019 Retrieved from STN Database accession no. 1998:513390 & TETRAHEDRON LETTERS vol. 39, no. 33, 1998, pages 6049 - 6052 *
DATABASE CAPLUS [Online] BEKHLI ET AL.: 'Synthesis of benzo(g)quinoline derivatives. I. 1,2,3,4-tetrahydro-4-oxobenzo(g)quinoline', XP002972027 Retrieved from STN Database accession no. 1969:106340 & KHIM GETEROTSIKL. SOEDIN., SB 1967, pages 296 - 299 *
DATABASE CAPLUS [Online] CRISAN ET AL.: 'Method of preparation of 1,4-dihydroxy-2-naphthoic acid', XP002972026 Retrieved from STN Database accession no. 1988:610742 & RO 93 222 A1 30 December 1987 *
DATABASE CAPLUS [Online] GOROG ET AL.: 'Herbicide antidote for plants', XP002972020 Retrieved from STN Database accession no. 1983:102692 & AT 368 678 B 25 October 1982 *
DATABASE CAPLUS [Online] HAHN: 'Arylamides of 2-naphthol-1-carboxylic acid', XP002972025 Retrieved from STN Database accession no. 1957:81756 & DE 838 290 C1 08 May 1952 *
DATABASE CAPLUS [Online] KAWABE ET AL.: 'Photosensitive composition of polymer having alicyclic backbone for positively working photoresist', XP002972021 Retrieved from STN Database accession no. 1999:781001 & JP 11 338 150 A 10 December 1999 *
DATABASE CAPLUS [Online] KAWANAKA ET AL.: 'Candida lipolytica HNA-110 strain and manufacture of 2,6-dihydroxy-3-naphthoic acid with candida species', XP002972029 Retrieved from STN Database accession no. 1998:55765 & JP 10 014 589 A 20 January 1998 *
DATABASE CAPLUS [Online] KOSUGI ET AL.: 'preparation of aromatic hydroxy carboxylic acids', XP002972031 Retrieved from STN Database accession no. 1999:409253 & JP 11 171 819 A 29 June 1999 *
DATABASE CAPLUS [Online] KUDARI ET AL.: 'Synthesis of a new series of 1,8-bis(5-aryloxymethyl-1,3,4-oxadiazol-2-y l) octanes and 1,8-bis(5-aryloxymethyl-4-amino/anilino-1,2 4-triazol-2-yl) octanes', XP002972022 Retrieved from STN Database accession no. 1998:125272 & ORIENTAL JOURNAL OF CHEMISTRY vol. 13, no. 3, 1997, pages 245 - 248 *
DATABASE CAPLUS [Online] LITVAK ET AL.: 'Substitution of an ester group for the halogen atom in 1-bromonaphthalene in the presence of molybdenum hexacarbonyl', XP002971471 Retrieved from STN Database accession no. 1980:550031 & ZHURNAL ORGANICHESKOI KHIMII vol. 16, no. 5, 1980, page 1097 *
DATABASE CAPLUS [Online] LU ET AL.: 'Synthesis of alkyl aryl ethers by phase transfer catalysis', XP002972024 Retrieved from STN Database accession no. 1996:158293 & HUAXUE SHIJI vol. 17, no. 6, 1995, pages 367 - 368 *
DATABASE CAPLUS [Online] MISHRA ET AL.: 'Use of some phenolic and pyrazolone derivatives as acid-base indicators', XP002972030 Retrieved from STN Database accession no. 1978:181806 & INDIAN JOURNAL OF CHEMISTRY vol. 15A, no. 10, 1977, pages 936 - 937 *
DATABASE CAPLUS [Online] NAKANISHI ET AL.: 'Process for controlling the isomer ratio of aromatic hydrocarboxylic acids in the carboxylation of alkali phenolates in phosphine oxide solvents', XP002972032 Retrieved from STN Database accession no. 1992:235255 & EP 0 478 197 A1 01 April 1992 *
DATABASE CAPLUS [Online] OSTAPCHUK ET AL.: 'Benzindoles. 24 Synthesis and certain properties of 5,6-benzotryptamine hydrochloride', XP002972028 Retrieved from STN Database accession no. 1984:454844 & KHIMIYA GETEROTSIKLICHESKIKH SOEDINENII no. 3, 1984, pages 346 - 348 *
DATABASE CAPLUS [Online] PAVLIDIS ET AL.: 'Synthesis of aliphatic amines related to monodansylcadaverine via the oxidative rearrangement of aliphati amides with phenyliodosyl bis(trifluoracetate)' Retrieved from STN Database accession no. 1989:172769 & SYNTHETIC COMMUNICATIONS vol. 18, no. 14, 1988, pages 1615 - 1624 *
DATABASE CAPLUS [Online] SEKINE ET AL.: 'Preparation of heterocycle-containing naphthoic acid derivatives as 5-lipoxygenase inhibitors and allergy inhibitors', XP002972033 Retrieved from STN Database accession no. 1993:603438 & JP 5 148 222 A 15 June 1993 *
DATABASE CAPLUS [Online] SHARMA ET AL.: 'Facile reduction of carboxylic acid imidazolides to primary alcohols in the presence of water', XP002972023 Retrieved from STN Database accession no. 1995:809508 & SYNLETT no. 8, 1995, pages 839 - 840 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2006117657A1 (fr) * 2005-05-03 2006-11-09 Ranbaxy Laboratories Limited Derives de triazolone utilises comme agents anti-inflammatoires

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