ABSTRACT
A series of thiomorpholine sulfonamide hydroxamate TACE inhibitors, all bearing propargylic ether P1' groups, was explored. In particular, compound 5h has excellent in vitro potency against isolated TACE enzyme and in cells, oral activity in a model of TNF-alpha production and a collagen-induced arthritis model, was selected as a clinical candidate for the treatment of RA.
Subject(s)
ADAM Proteins/antagonists & inhibitors , Acetylene/chemistry , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/pharmacology , ADAM17 Protein , Administration, Oral , Alkynes/chemistry , Animals , Arthritis/drug therapy , Caco-2 Cells , Collagen/toxicity , Crystallography, X-Ray , Disease Models, Animal , Dogs , Haplorhini , Humans , Hydroxamic Acids/chemistry , Lipopolysaccharides/pharmacology , Matrix Metalloproteinase 13 , Matrix Metalloproteinase Inhibitors , Mice , Molecular Structure , Morpholines/chemistry , Propanols/chemistry , Rats , Structure-Activity Relationship , Sulfonamides/chemistry , Tumor Necrosis Factor-alpha/metabolismABSTRACT
The SAR of a series of potent sulfonamide hydroxamate TACE inhibitors bearing a butynyloxy P1' group was explored. In particular, compound 5k has excellent in vitro potency against TACE enzyme and in cells, and oral activity in an in vivo model of TNF-alpha production and a collagen-induced arthritis model.
Subject(s)
ADAM Proteins/antagonists & inhibitors , Cysteine Proteinase Inhibitors/chemical synthesis , Hydroxamic Acids/chemical synthesis , Sulfonamides/chemical synthesis , ADAM17 Protein , Animals , Biological Availability , Caspase Inhibitors , Cell Line , Crystallography, X-Ray , Cysteine Proteinase Inhibitors/pharmacokinetics , Cysteine Proteinase Inhibitors/pharmacology , Dogs , Humans , Hydroxamic Acids/pharmacokinetics , Hydroxamic Acids/pharmacology , Inhibitory Concentration 50 , Lipopolysaccharides/pharmacology , Mice , Molecular Structure , Monocytes , Rats , Species Specificity , Structure-Activity Relationship , Sulfonamides/pharmacokinetics , Sulfonamides/pharmacology , Tumor Necrosis Factor-alpha/biosynthesis , Tumor Necrosis Factor-alpha/drug effectsABSTRACT
The SAR of a series of potent sulfonamide hydroxamate TACE inhibitors, all bearing a butynyloxy P1' group, was explored. In particular, compound 5j has excellent in vitro potency against isolated TACE enzyme and in cells, good selectivity over MMP-1 and MMP-9, and oral activity in an in vivo model of TNF-alpha production and a collagen-induced arthritis model.
Subject(s)
Acetylene/chemistry , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/pharmacology , Metalloendopeptidases/antagonists & inhibitors , ortho-Aminobenzoates/chemistry , ADAM Proteins , ADAM17 Protein , Crystallography, X-Ray , Hydroxamic Acids/chemistry , Molecular Structure , Structure-Activity Relationship , Sulfonamides/chemistry , ortho-Aminobenzoates/pharmacologyABSTRACT
Potent and selective bicyclic heteroaryl hydroxamic acid MMP and TACE inhibitors were synthesized by a novel convergent route. Selectivity and efficacy versus MMPs and TACE could be controlled by appropriate substitution on the scaffolds and by variation of the P1' group. Select compounds were found to be effective in in vivo models of arthritis.
Subject(s)
Enzyme Inhibitors/chemistry , Enzyme Inhibitors/pharmacology , Hydroxamic Acids/chemistry , Hydroxamic Acids/pharmacology , Matrix Metalloproteinase Inhibitors , Metalloendopeptidases/antagonists & inhibitors , ADAM Proteins , ADAM17 Protein , Animals , Arthritis/drug therapy , Arthritis/pathology , Biological Availability , Bridged Bicyclo Compounds, Heterocyclic/chemical synthesis , Bridged Bicyclo Compounds, Heterocyclic/chemistry , Bridged Bicyclo Compounds, Heterocyclic/pharmacology , Cartilage/drug effects , Cartilage/pathology , Cattle , Disease Models, Animal , Enzyme Inhibitors/chemical synthesis , Hydroxamic Acids/chemical synthesis , Mice , Rats , Structure-Activity RelationshipABSTRACT
The SAR of a series of potent sulfonamide hydroxamate TACE inhibitors bearing novel acetylenic P1' groups was explored. In particular, compound 4t bearing a butynyloxy P1' moiety has excellent in vitro potency against isolated TACE enzyme and in cells, good selectivity over MMP-1 and oral activity in an in vivo model of TNF-alpha production.
Subject(s)
Acetylene/chemistry , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/pharmacology , Hydroxamic Acids/chemistry , Hydroxamic Acids/pharmacology , Metalloendopeptidases/antagonists & inhibitors , Sulfonamides/chemical synthesis , Sulfonamides/pharmacology , ortho-Aminobenzoates/chemistry , ortho-Aminobenzoates/pharmacology , ADAM Proteins , ADAM17 Protein , Structure-Activity RelationshipABSTRACT
Anthranilic acid derivatives bearing basic amines were prepared and evaluated in vitro and in vivo as inhibitors of MMP-1, MMP-9, MMP-13, and TACE. Piperazine 4u has been identified as a potent, selective, orally active inhibitor of MMP-9 and MMP-13.
Subject(s)
Amines/chemistry , Enzyme Inhibitors/pharmacology , Matrix Metalloproteinase Inhibitors , ortho-Aminobenzoates/pharmacology , ADAM Proteins , ADAM17 Protein , Animals , Binding Sites , Collagenases/metabolism , Enzyme Inhibitors/chemical synthesis , Enzyme Inhibitors/chemistry , Hydroxamic Acids/chemistry , Interleukin-1 , Interleukin-1beta , Magnetic Resonance Spectroscopy , Matrix Metalloproteinase 1/metabolism , Matrix Metalloproteinase 13 , Matrix Metalloproteinase 9/metabolism , Metalloendopeptidases , Mice , Models, Molecular , Osteoarthritis/drug therapy , Rats , Structure-Activity Relationship , ortho-Aminobenzoates/chemical synthesis , ortho-Aminobenzoates/chemistryABSTRACT
BACKGROUND: Transforming growth factor-alpha (TGF-alpha) expression is abnormal in polycystic kidney disease. We previously demonstrated that blockade of the epidermal growth factor receptor (EGFR), the receptor for TGF-alpha, significantly slowed disease progression in the bpk murine model of autosomal-recessive kidney disease (ARPKD). In the present study, kidney TGF-alpha expression in this model is characterized, and the therapeutic potential of inhibiting TGF-alpha in ARPKD is examined using a novel inhibitor of tumor necrosis factor-alpha converting enzyme (TACE), the metalloproteinase that cleaves membrane-bound TGF-alpha to release the secreted ligand. METHODS: Immunohistochemistry (IH) and Western analysis were performed on kidneys from cystic bpk mice and noncystic littermates at postnatal days 7, 14, and 21. Bpk mice and normal controls were treated with WTACE2, a competitive inhibitor of TACE, from day 7 until day 21, and the effects on kidney histology and renal function were assessed. RESULTS: Increased TGF-alpha expression by IH was demonstrated in the proximal tubules (PT) at postnatal day 7 and collecting tubules (CT) by day 21. A parallel increase in kidney TGF-alpha expression was demonstrated by Western analysis. Treatment of cystic bpk mice with WTACE2 resulted in a 43% reduction in kidney weight to body weight ratio (11.2 vs. 19.7%), improved cystic index (3.2 vs. 4.8), reduced cystic CT to PT ratio (1.2 vs. 8), and a greater than 30% reduction in BUN and serum creatinine. CONCLUSIONS: These findings support the pathophysiological role of the TGF-alpha/EGFR axis in murine ARPKD and demonstrate that inhibition of TGF-alpha secretion has therapeutic potential in PKD.
Subject(s)
Hydroxamic Acids/therapeutic use , Metalloendopeptidases/antagonists & inhibitors , Polycystic Kidney, Autosomal Recessive/drug therapy , Sulfonamides/therapeutic use , ADAM Proteins , ADAM17 Protein , Aging/metabolism , Animals , Animals, Newborn/growth & development , Animals, Newborn/metabolism , Immunohistochemistry , Kidney/metabolism , Kidney/pathology , Mice , Mice, Inbred Strains , Polycystic Kidney, Autosomal Recessive/metabolism , Polycystic Kidney, Autosomal Recessive/pathology , Transforming Growth Factor alpha/metabolismABSTRACT
A novel series of anthranilic acid-based inhibitors of MMP-1, MMP-9, MMP-13, and TACE was prepared and evaluated. Selective inhibitors of MMP-9, MMP-13, and TACE were identified, including the potent, orally active MMP-13 inhibitor 4p.
Subject(s)
Enzyme Inhibitors/pharmacology , Matrix Metalloproteinase Inhibitors , ortho-Aminobenzoates/pharmacology , ADAM Proteins , ADAM17 Protein , Collagenases/metabolism , Enzyme Inhibitors/chemical synthesis , Enzyme Inhibitors/chemistry , Matrix Metalloproteinase 13 , Matrix Metalloproteinase 9/metabolism , Matrix Metalloproteinases/metabolism , Metalloendopeptidases/antagonists & inhibitors , Metalloendopeptidases/metabolism , Structure-Activity Relationship , ortho-Aminobenzoates/chemical synthesis , ortho-Aminobenzoates/chemistryABSTRACT
Heteroaryl and cycloalkyl sulfonamide-hydroxamic acid MMP inhibitors were investigated. Of these, the pyridyl analogue 2 is the most potent and selective inhibitor of MMP-9 and MMP-13 in vitro.
Subject(s)
Hydroxamic Acids/pharmacology , Matrix Metalloproteinase Inhibitors , Protease Inhibitors/chemical synthesis , ortho-Aminobenzoates/pharmacology , Animals , Combinatorial Chemistry Techniques , Humans , Hydroxamic Acids/chemical synthesis , Hydroxamic Acids/chemistry , Inhibitory Concentration 50 , Matrix Metalloproteinase 13 , Protease Inhibitors/chemistry , Protease Inhibitors/pharmacology , Structure-Activity Relationship , Sulfonamides/chemical synthesis , Sulfonamides/chemistry , Sulfonamides/pharmacology , ortho-Aminobenzoates/chemical synthesis , ortho-Aminobenzoates/chemistryABSTRACT
A novel series of anthranilic acid-based inhibitors of MMP-1, MMP-9, and MMP-13 was prepared and evaluated both in vitro and in vivo. The most potent compound, 6e, has in vivo activity in a rat sponge-wrapped cartilage model.
Subject(s)
Matrix Metalloproteinase Inhibitors , Protease Inhibitors/chemical synthesis , ortho-Aminobenzoates/pharmacology , Animals , Combinatorial Chemistry Techniques , Humans , Hydroxamic Acids/chemical synthesis , Hydroxamic Acids/chemistry , Hydroxamic Acids/pharmacology , Inhibitory Concentration 50 , Matrix Metalloproteinase 13 , Protease Inhibitors/chemistry , Protease Inhibitors/pharmacology , Structure-Activity Relationship , Sulfonamides/chemical synthesis , Sulfonamides/chemistry , Sulfonamides/pharmacology , ortho-Aminobenzoates/chemical synthesis , ortho-Aminobenzoates/chemistryABSTRACT
The high-resolution solution structure of the catalytic fragment of human collagenase-3 (MMP-13) complexed with a sulfonamide derivative of a hydroxamic acid compound (WAY-151693) has been determined by multidimensional heteronuclear NMR. A total of 30 structures were calculated for residues 7-164 by means of hybrid distance geometry-simulated annealing using a total of 3280 experimental NMR restraints. The atomic rms distribution about the mean coordinate positions for the 30 structures is 0.43(+/-0.05) A for the backbone atoms, 0.80(+/-0.09) A for all atoms, and 0.47(+/-0.04) A for all atoms excluding disordered side-chains. The overall structure of MMP-13 is composed of a beta-sheet consisting of five beta-strands in a mixed parallel and anti-parallel arrangement and three alpha-helices where its overall fold is consistent with previously solved MMP structures. A comparison of the NMR structure of MMP-13 with the published 1.6 A resolution X-ray structure indicates that the major differences between the structures is associated with loop dynamics and crystal-packing interactions. The side-chains of some active-site residues for the NMR and X-ray structures of MMP-13 adopt distinct conformations. This is attributed to the presence of unique inhibitors in the two structures that encounter distinct interactions with MMP-13. The major structural difference observed between the MMP-13 and MMP-1 NMR structures is the relative size and shape of the S1' pocket where this pocket is significantly longer for MMP-13, nearly reaching the surface of the protein. Additionally, MMP-1 and MMP-13 exhibit different dynamic properties for the active-site loop and the structural Zn-binding region. The inhibitor WAY-151693 is well defined in the MMP-13 active-site based on a total of 52 distance restraints. The binding motif of WAY-151693 in the MMP-13 complex is consistent with our previously reported MMP-1:CGS-27023A NMR structure and is similar to the MMP-13: RS-130830 X-ray structure.
Subject(s)
Catalytic Domain , Collagenases/chemistry , Collagenases/metabolism , Hydroxamic Acids/metabolism , Protease Inhibitors/metabolism , Pyrazines , Amino Acid Sequence , Binding Sites , Crystallography, X-Ray , Humans , Hydroxamic Acids/chemistry , Hydroxamic Acids/pharmacology , Inhibitory Concentration 50 , Matrix Metalloproteinase 1/chemistry , Matrix Metalloproteinase 1/metabolism , Matrix Metalloproteinase 13 , Matrix Metalloproteinase Inhibitors , Models, Molecular , Molecular Sequence Data , Nuclear Magnetic Resonance, Biomolecular , Peptide Fragments/chemistry , Peptide Fragments/metabolism , Protease Inhibitors/chemistry , Protease Inhibitors/pharmacology , Protein Structure, Secondary , Sequence Alignment , Sequence Homology, Amino Acid , Solutions , Sulfonamides , Zinc/metabolismSubject(s)
Collagenases/chemistry , Hydroxamic Acids/chemistry , Nuclear Magnetic Resonance, Biomolecular , Protein Structure, Secondary , Sulfonamides/chemistry , Enzyme Stability , Humans , Hydroxamic Acids/antagonists & inhibitors , Hydroxamic Acids/metabolism , Matrix Metalloproteinase 13 , Molecular Structure , Protein Binding , Recombinant Proteins/chemistry , Sulfonamides/metabolismABSTRACT
Experimental evidence confirms that the matrix metalloproteinases (MMPs) play a fundamental role in a wide variety of pathologic conditions that involve connective tissue destruction including osteoarthritis and rheumatoid arthritis, tumor metastasis and angiogenesis, corneal ulceration, multiple sclerosis, periodontal disease, and atherosclerosis. Modulation of MMP regulation is possible at several biochemical sites, but direct inhibition of enzyme action provides a particularly attractive target for therapeutic intervention. Hypotheses concerning inhibition of specific MMP(s) with respect to disease target and/or side-effect profile have emerged. Examples are presented of recent advances in medicinal chemistry approaches to the design of matrix metalloproteinase inhibitors (MMPIs), approaches that address structural requirements and that influence potency, selectivity, and bioavailability. Two important approaches to the design, synthesis, and biological evaluation of MMPIs are highlighted: (1) the invention of alternatives to hydroxamic acid zinc chelators and (2) the construction of nonpeptide scaffolds. One current example in each of these two approaches from our own work is described.
Subject(s)
Enzyme Inhibitors/chemical synthesis , Metalloendopeptidases/antagonists & inhibitors , Animals , Arteriosclerosis/drug therapy , Arteriosclerosis/pathology , Arteriosclerosis/physiopathology , Arthritis, Rheumatoid/drug therapy , Arthritis, Rheumatoid/pathology , Arthritis, Rheumatoid/physiopathology , Connective Tissue/enzymology , Connective Tissue/pathology , Drug Design , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/therapeutic use , Extracellular Matrix/enzymology , Humans , Neoplasm Metastasis , Osteoarthritis/drug therapy , Osteoarthritis/pathology , Osteoarthritis/physiopathology , Periodontal Diseases/drug therapy , Periodontal Diseases/pathology , Periodontal Diseases/physiopathologyABSTRACT
A novel series of matrix metalloproteinase (MMP) inhibitors is described. Incorporation of a terminal alpha-mercaptoketone or alpha-mercaptoalcohol in the zinc binding domain of a series of inhibitors led to compounds exhibiting low nanomolar activity against collagenase-1 (MMP-1), stromelysin (MMP-3), and gelatinase-B (MMP-9).
Subject(s)
Alcohols/chemistry , Ketones/chemistry , Metalloendopeptidases/antagonists & inhibitors , Protease Inhibitors/chemistry , Sulfhydryl Compounds/chemistry , Alcohols/chemical synthesis , Alcohols/pharmacology , Binding Sites , Drug Design , Hydroxamic Acids/chemistry , Hydroxamic Acids/pharmacology , Ketones/chemical synthesis , Ketones/pharmacology , Kinetics , Matrix Metalloproteinase 1 , Matrix Metalloproteinase 9 , Matrix Metalloproteinase Inhibitors , Molecular Structure , Protease Inhibitors/chemical synthesis , Protease Inhibitors/pharmacology , Structure-Activity Relationship , Sulfhydryl Compounds/chemical synthesis , Sulfhydryl Compounds/pharmacology , Zinc/metabolismABSTRACT
A series of succinyl based mercaptoketones and diastereomeric mercaptoalcohols were prepared and evaluated in vitro as inhibitors of the matrix metalloproteinases collagenase-1 (MMP-1), stromelysin (MMP-3), and gelatinase-B (MMP-9).
Subject(s)
Alcohols/chemical synthesis , Ketones/chemical synthesis , Metalloendopeptidases/antagonists & inhibitors , Protease Inhibitors/chemical synthesis , Succinates/chemical synthesis , Sulfhydryl Compounds/chemical synthesis , Alcohols/chemistry , Alcohols/pharmacology , Drug Design , Indicators and Reagents , Ketones/chemistry , Ketones/pharmacology , Kinetics , Matrix Metalloproteinase 1 , Matrix Metalloproteinase 9 , Matrix Metalloproteinase Inhibitors , Molecular Structure , Protease Inhibitors/chemistry , Protease Inhibitors/pharmacology , Structure-Activity Relationship , Succinates/chemistry , Succinates/pharmacology , Sulfhydryl Compounds/chemistry , Sulfhydryl Compounds/pharmacologyABSTRACT
A novel series of diazepine-based hydroxamic acid inhibitors of MMP-1, MMP-9, and MMP-13 were prepared and evaluated both in vitro and in vivo.
Subject(s)
Azepines/metabolism , Enzyme Inhibitors/chemical synthesis , Enzyme Inhibitors/pharmacology , Matrix Metalloproteinase Inhibitors , Organic Chemicals , Pyrazines , Animals , Antineoplastic Agents/pharmacology , Hydroxamic Acids/chemical synthesis , Hydroxamic Acids/pharmacology , Matrix Metalloproteinase 1 , Matrix Metalloproteinase 9 , Mice , Protease Inhibitors/pharmacology , Structure-Activity Relationship , Sulfonamides/chemical synthesis , Sulfonamides/pharmacologyABSTRACT
Alpha receptor and angiotensin II blockade were used to study renal cortical vasoconstrictor mechanisms during hemorrhage to 70 mmHg in chloralose-urethane anesthetized dogs. A freeze-dissection 133Xe disappearance technique was utilized to assess renal blood flow patterns. Hemorrhage alone caused a 30% decrease in total renal blood flow (TRBF) and a 50% decrease in outer cortical blood flow. Inner cortical flow decreased approximately 30%. Outer medullary blood flow decreased 25%. Renal arterial infusion of phentolamine beginning 20 min posthemorrhage produced no alteration in the expected posthemorrhage TRBF or its distribution. Plasma renin activity increased 6-7 fold in the hemorrhage group as well as in the hemorrhage with phentolamine group. Saralasin treatment beginning 20 min posthemorrhage produced a pattern in which neither TRBF nor cortical blood flow was significantly reduced by the hemorrhage. It therefore appears that angiotensin II is a renal cortical vasoconstrictor during hemorrhage. These data demonstrate that during the first 30 minutes of hemorrhage the renal cortical vasoconstriction is mediated by angiotensin II and appears to be independent of alpha-adrenergic mechanisms.
Subject(s)
Angiotensin II/physiology , Hemorrhage/physiopathology , Receptors, Adrenergic, alpha/physiology , Receptors, Adrenergic/physiology , Renal Circulation , Angiotensin II/antagonists & inhibitors , Animals , Dogs , Kidney Cortex/blood supply , Phentolamine/pharmacology , Receptors, Adrenergic, alpha/drug effects , Renin-Angiotensin System , Saralasin/pharmacology , VasoconstrictionABSTRACT
Dopamine has been reported to increase renal blood flow during hemorrhagic shock. Since this effect would be of considerable benefit in shock, the effect of dopamine on hemorrhagic hypotension to 70 mm Hg for five hours was studied. Plasma renin activity and outer cortical blood flow were significantly greater in the group of dogs receiving dopamine. Total renal blood flow, sodium excretion, and potassium excretion were similar in both groups; however, the ratio of urine sodium to potassium concentration followed closely the plasma renin activity. We conclude that dopamine infusion alone is of limited value in improving renal hemodynamics and function during hemorrhagic hypotension to 70 mm Hg. In addition, increased plasma renin activity produced by dopamine infusion during hemorrhage would tend to offset the expected increases in renal blood flow and sodium excretion.