ABSTRACT
Inhibition of hydroxy acid oxidase 1 (HAO1) is a strategy to mitigate the accumulation of toxic oxalate that results from reduced activity of alanine-glyoxylate aminotransferase (AGXT) in primary hyperoxaluria 1 (PH1) patients. DNA-Encoded Chemical Library (DECL) screening provided two novel chemical series of potent HAO1 inhibitors, represented by compounds 3-6. Compound 5 was further optimized via various structure-activity relationship (SAR) exploration methods to 29, a compound with improved potency and absorption, distribution, metabolism, and excretion (ADME)/pharmacokinetic (PK) properties. Since carboxylic acid-containing compounds are often poorly permeable and have potential active glucuronide metabolites, we undertook a brief, initial exploration of acid replacements with the aim of identifying non-acid-containing HAO1 inhibitors. Structure-based drug design initiated with Compound 5 led to the identification of a nonacid inhibitor of HAO1, 31, which has weaker potency and increased permeability.
Subject(s)
Alcohol Oxidoreductases/antagonists & inhibitors , DNA/chemistry , Small Molecule Libraries/chemistry , Alcohol Oxidoreductases/metabolism , Animals , Binding Sites , Crystallography, X-Ray , DNA/metabolism , Drug Design , Half-Life , Humans , Hyperoxaluria, Primary/metabolism , Hyperoxaluria, Primary/pathology , Indoles/chemistry , Indoles/metabolism , Male , Mice , Molecular Docking Simulation , Small Molecule Libraries/metabolism , Structure-Activity Relationship , Thiazoles/chemistry , Thiazoles/metabolism , Transaminases/genetics , Transaminases/metabolismABSTRACT
Design, synthesis and structure-activity relationship of a series of biphenylsulfonamido-3-methylbutanoic acid based aggrecanase-1 inhibitors are described. In addition to robust aggrecanase-1 inhibition, these compounds also exhibit potent MMP-13 activity. In cell-based cartilage explants assay compound 48 produced 87% inhibition of proteoglycan degradation at 10 µg/mL. Good pharmacokinetic properties were demonstrated by 46 with a half-life of 6h and bioavailability of 23%.
Subject(s)
ADAM Proteins/antagonists & inhibitors , ADAM Proteins/metabolism , Biphenyl Compounds/pharmacology , Procollagen N-Endopeptidase/antagonists & inhibitors , Procollagen N-Endopeptidase/metabolism , Protease Inhibitors/pharmacology , Sulfonamides/pharmacology , ADAMTS4 Protein , Animals , Biphenyl Compounds/chemistry , Biphenyl Compounds/pharmacokinetics , Drug Design , Humans , Male , Matrix Metalloproteinase 13/metabolism , Models, Molecular , Osteoarthritis/drug therapy , Protease Inhibitors/chemistry , Protease Inhibitors/pharmacokinetics , Proteoglycans/metabolism , Rats , Rats, Sprague-Dawley , Structure-Activity Relationship , Sulfonamides/chemistry , Sulfonamides/pharmacokineticsABSTRACT
The prevention of aggrecan (a key component of cartilage) cleavage via the inhibition of aggrecanase-1 may provide a unique opportunity to stop the progression of cartilage degradation in osteoarthritis. The evaluation of a series of biphenylsulfonamides resulted in the identification of the ((4-keto)-phenoxy)methyl biphenyl-4-sulfonamides analogs (19-21 and 24) with improved Agg-1 inhibition and MMP-2, MMP-13 activity.
Subject(s)
ADAM Proteins/antagonists & inhibitors , ADAM Proteins/metabolism , Chemistry, Pharmaceutical/methods , Osteoarthritis/drug therapy , Procollagen N-Endopeptidase/antagonists & inhibitors , Procollagen N-Endopeptidase/metabolism , Sulfonamides/chemical synthesis , ADAMTS4 Protein , Cartilage/drug effects , Cartilage/metabolism , Drug Design , Humans , Inhibitory Concentration 50 , Matrix Metalloproteinase 13/metabolism , Matrix Metalloproteinase 2/metabolism , Models, Chemical , Molecular Conformation , Proteoglycans/chemistry , Sulfonamides/pharmacologyABSTRACT
N-((8-Hydroxy-5-substituted-quinolin-7-yl)(phenyl)methyl)-2-phenyloxy/amino-acetamide inhibitors of ADAMTS-5 (Aggrecanase-2) have been prepared. Selected compounds 10, 14, 25, and 53 show sub-microM ADAMTS-5 potency and good selectivity over the related metalloproteases ADAMTS-4 (Aggrecanase-1), MMP-13, and MMP-12. Compound 53 shows a good balance of potent ADAMTS-5 inhibition, moderate CYP3A4 inhibition and good rat liver microsome stability. This series of compounds represents progress towards selective ADAMTS-5 inhibitors as disease modifying osteoarthritis agents.
Subject(s)
ADAM Proteins/antagonists & inhibitors , ADAM Proteins/chemistry , Acetamides/chemical synthesis , Acetamides/pharmacology , ADAM Proteins/metabolism , ADAMTS4 Protein , ADAMTS5 Protein , Animals , Chemistry, Pharmaceutical/methods , Cytochrome P-450 CYP3A , Cytochrome P-450 CYP3A Inhibitors , Cytochrome P-450 Enzyme Inhibitors , Drug Design , Humans , Inhibitory Concentration 50 , Microsomes, Liver/drug effects , Models, Chemical , Osteoarthritis/drug therapy , Procollagen N-Endopeptidase/metabolism , RatsABSTRACT
Aggrecanases are now believed to be the principal proteinases responsible for aggrecan degradation in osteoarthritis. Given their potential as a drug target, we solved crystal structures of the two most active human aggrecanase isoforms, ADAMTS4 and ADAMTS5, each in complex with bound inhibitor and one wherein the enzyme is in apo form. These structures show that the unliganded and inhibitor-bound enzymes exhibit two essentially different catalytic-site configurations: an autoinhibited, nonbinding, closed form and an open, binding form. On this basis, we propose that mature aggrecanases exist as an ensemble of at least two isomers, only one of which is proteolytically active.
Subject(s)
ADAM Proteins/chemistry , Procollagen N-Endopeptidase/chemistry , ADAMTS4 Protein , ADAMTS5 Protein , Binding Sites , Crystallography, X-Ray , Enzyme Inhibitors/chemistry , Humans , Models, Molecular , Protein ConformationABSTRACT
5'-Phenyl-3'H-spiro[indoline-3,2'-[1,3,4]thiadiazol]-2-one inhibitors of ADAMTS-5 (Aggrecanase-2) have been prepared via commercially available starting materials. Selected compounds 23, 33-35 show sub-micromolar ADAMTS-5 potency and strong SAR trends with selectivity over the related metalloproteases ADAMTS-4 (Aggrecanase-1), MMP12, and MMP13. This series of compounds represents progress toward a selective ADAMTS-5 inhibitor as a disease modifying osteoarthritis drug.
Subject(s)
Endopeptidases/metabolism , Indoles/chemistry , Protease Inhibitors/chemistry , Protease Inhibitors/pharmacology , Spiro Compounds/chemistry , Spiro Compounds/pharmacology , Thiadiazoles/chemistry , Molecular Structure , Protease Inhibitors/chemical synthesis , Spiro Compounds/chemical synthesis , Structure-Activity Relationship , Thiadiazoles/chemical synthesis , Thiadiazoles/pharmacologyABSTRACT
5-Benzylidene-2-thioxo-thiazolidin-4-one inhibitors of ADAMTS-5 (Aggrecanase-2) have been prepared via commercially available starting materials. The identified compounds show micromolar ADAMTS-5 potency and demonstrate SAR trends for both the aryl group and thioxothiazolidinone zinc chelator. This series of compounds represents steps toward a metalloprotease inhibitor as a disease-modifying osteoarthritis drug.
Subject(s)
ADAM Proteins/antagonists & inhibitors , Thiazolidinediones/chemical synthesis , Thiazolidinediones/pharmacology , ADAMTS5 Protein , Chelating Agents , Enzyme Inhibitors/pharmacology , Enzyme Inhibitors/therapeutic use , Humans , Metalloproteases/antagonists & inhibitors , Osteoarthritis/drug therapy , Structure-Activity Relationship , ZincABSTRACT
A series of 5-((1H-pyrazol-4-yl)methylene)-2-thioxothiazolidin-4-one inhibitors of ADAMTS-5 (aggrecanase-2) is described. These compounds show microM functional inhibition of ADAMTS-5, and represent a new class of agents with the potential of inhibiting degradation of aggrecan, a major component of cartilage which is lost in osteoporosis. Compound 12 is noteworthy in that it has an ADAMTS-5 IC50: 1.1 microM and shows >40-fold functional selectivity over ADAMTS-4 (aggrecanase-1).
Subject(s)
ADAM Proteins/antagonists & inhibitors , Thiazolidinediones/chemical synthesis , ADAMTS4 Protein , ADAMTS5 Protein , Aggrecans/drug effects , Aggrecans/metabolism , Cartilage , Humans , Inhibitory Concentration 50 , Osteoporosis/drug therapy , Procollagen N-Endopeptidase , Structure-Activity Relationship , Thiazolidinediones/pharmacologyABSTRACT
Aggrecanases are recently discovered enzymes that cleave aggrecan, a key component of cartilage. Aggrecanase inhibitors may provide a unique means to halt the progression of cartilage destruction in osteoarthritis. The synthesis and evaluation of biphenylsulfonamidocarboxylic acid inhibitors of aggrecanase-1 are reported. Compound 24 demonstrated 89% inhibition of proteoglycan degradation at 10 microg/mL and has an oral bioavailability in rat of 35%.
Subject(s)
ADAM Proteins/antagonists & inhibitors , Biphenyl Compounds/chemistry , Carboxylic Acids , Enzyme Inhibitors , Procollagen N-Endopeptidase/antagonists & inhibitors , Sulfonamides/chemistry , ADAMTS4 Protein , Administration, Oral , Animals , Carboxylic Acids/chemical synthesis , Carboxylic Acids/chemistry , Carboxylic Acids/pharmacology , Collagenases/metabolism , Crystallography, X-Ray , Drug Evaluation, Preclinical , Enzyme Inhibitors/chemical synthesis , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/pharmacology , Matrix Metalloproteinase 13 , Matrix Metalloproteinase Inhibitors , Models, Molecular , Molecular Structure , Proteoglycans/drug effects , Proteoglycans/metabolism , Rats , Structure-Activity RelationshipABSTRACT
Members of the ADAMTS (a disintegrin and metalloprotease with thrombospondin motifs) family share common structural features including a disintegrin domain, a zinc metalloprotease domain, and at least one thrombospondin motif. Aberrant expression of several of these proteins has led to an understanding of their role in human disease; however, a link to function for many has not yet been made. One such uncharacterized family member, ADAMTS-8, shares significant protein sequence homology with a subgroup of ADAMTSs that includes ADAMTS-1, ADAMTS-4, ADAMTS-5, and ADAMTS-15. Each of these proteases has been shown to cleave 'aggrecanase-susceptible' site(s) within the extracellular matrix (ECM) proteoglycan aggrecan, and ADAMTS-4 and ADAMTS-5 have been postulated to play a role in the depletion of articular cartilage in osteoarthritic disease. Based on sequence relationships, in the present study we examined the ability of ADAMTS-8 to exhibit 'aggrecanase' activity. A neoepitope monoclonal antibody (MAb; AGG-C1; anti-NITEGE373) was developed and used to demonstrate the ability of ADAMTS-8 to cleave aggrecan at the aggrecanase-susceptible Glu373-Ala374 peptide bond. In addition, expression analyses demonstrated the presence of ADAMTS-8 mRNA transcripts in normal and osteoarthritic human cartilage.