ABSTRACT
BACKGROUND: Vitrification by Rapid-I method could be essential for felid rescue programs to protect wild felid in the future. OBJECTIVE: This study was aimed at adapting the Rapid I method and evaluating the viability of serval and Pallas cat oocytes compared to oocytes of the domestic cat. MATERIALS AND METHODS: Oocytes after collection and in vitro maturation were vitrified using Cryotech medium (Cryotech, Japan) and a Rapid-I device (Vitrolife, Sweden). To evaluate viability, oocytes after warming were stained with fluorescein diacetate and ethidium bromide. RESULTS: Survival rate in the control group (domestic cat) was 75 %. In the experimental group, 70% (serval) and 60% (pallas cat) viable oocytes were found. CONCLUSION: The Rapid-I method can be applied successfully for the vitrification of wild felid oocytes.
Subject(s)
Cell Survival , Cryopreservation/veterinary , Felidae , Felis , Oocytes/cytology , Animals , Cryopreservation/methods , Cryoprotective Agents , Female , VitrificationABSTRACT
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
The mammalian target of rapamycin (mTOR) is a central regulator of G1 cell cycle protein synthesis that precedes commitment to normal cellular replication. We have studied the effect of cell cycle inhibitor-779 (CCI-779), a rapamycin ester that inhibits mTOR function, on the proliferation of a panel of breast cancer cell lines. Six of eight lines studied were sensitive (IC(50)< or = 50 nM) and two lines were resistant (IC(50)>1.0 microM) to CCI-779. Sensitive lines were estrogen dependent (MCF-7, BT-474, T-47D), or lacked expression of the tumor suppressor PTEN (MDA-MB-468, BT-549), and/or overexpressed the Her-2/neu oncogene (SKBR-3, BT-474). Resistant lines (MDA-MB-435, MDA-MB-231) shared none of these properties. CCI-779 (50 nM) inhibited mTOR function in both a sensitive and a resistant line. In nu/nu mouse xenografts, CCI-779 inhibited growth of MDA-MB-468 (sensitive) but not MDA-MB-435 resistant tumors. Treatment of sensitive lines with CCI-779 resulted in a decrease in D-type cyclin and c-myc levels and an increase in p27(kip-1) levels. There was good correlation between activation of the Akt pathway and sensitivity to CCI-779. Amplification of mTOR-regulated p70S6 kinase, which is downstream of Akt, may also have conferred CCI-779 sensitivity to MCF-7 cells. Taken together, the data suggest that mTOR may be a good target for breast cancer therapy, especially in tumors with Akt activation resulting from either growth factor dependency or loss of PTEN function.
Subject(s)
Antineoplastic Agents/pharmacology , Breast Neoplasms/drug therapy , Protein Kinase Inhibitors , Protein Kinases , Sirolimus/analogs & derivatives , Sirolimus/pharmacology , Animals , Antineoplastic Agents/therapeutic use , Breast Neoplasms/metabolism , Breast Neoplasms/pathology , Drug Evaluation, Preclinical , Drug Resistance, Neoplasm , Female , Mice , Mice, Nude , Sirolimus/therapeutic use , TOR Serine-Threonine Kinases , Tumor Cells, Cultured/drug effectsABSTRACT
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
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
The solution structure of the catalytic fragment of human fibroblast collagenase (MMP-1) complexed with a sulfonamide derivative of a hydroxamic acid compound (CGS-27023A) has been determined using two-dimensional and three-dimensional heteronuclear NMR spectroscopy. The solution structure of the complex was calculated by means of hybrid distance geometry-simulated annealing using a combination of experimental NMR restraints obtained from the previous refinement of the inhibitor-free MMP-1 (1) and recent restraints for the MMP-1:CGS-27023A complex. The hydroxamic acid moiety of CGS-27023A was found to chelate to the "right" of the catalytic zinc where the p-methoxyphenyl sits in the S1' active-site pocket, the isopropyl group is in contact with H83 and N80, and the pyridine ring is solvent exposed. The sulfonyl oxygens are in hydrogen-bonding distance to the backbone NHs of L81 and A82. This is similar to the conformation determined by NMR of the inhibitor bound to stromelysin (2, 3). A total of 48 distance restraints were observed between MMP-1 and CGS-27023A from 3D 13C-edited/12C-filtered NOESY and 3D 15N-edited NOESY experiments. An additional 18 intramolecular restraints were observed for CGS-27023A from a 2D 12C-filtered NOESY experiment. A minimal set of NMR experiments in combination with the free MMP-1 assignments were used to assign the MMP-1 (1)H, 13C, and 15N resonances in the MMP-1:CGS-27023A complex. The assignments of CGS-27023A in the complex were obtained from 2D 12C-filtered NOESY and 2D 12C-filtered TOCSY experiments.
Subject(s)
Collagenases/chemistry , Fibroblasts/enzymology , Hydroxamic Acids/chemistry , Peptide Fragments/chemistry , Protease Inhibitors/chemistry , Pyrazines , Sulfonamides/chemistry , Catalysis , Computer Simulation , Crystallography, X-Ray , Humans , Macromolecular Substances , Matrix Metalloproteinase 1 , Matrix Metalloproteinase 3/chemistry , Matrix Metalloproteinase 8 , Matrix Metalloproteinase Inhibitors , Models, Molecular , Nuclear Magnetic Resonance, Biomolecular , Peptide Fragments/antagonists & inhibitors , Protein Conformation , SolutionsABSTRACT
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
The effects of potential anti-osteoarthritic compounds both on the direct inhibition of collagenase and neutral protease activities and on IL-1 induced release of neutral proteases from rabbit articular chondrocytes were investigated. WY-46,135 ((+)-N-[[[(5-chloro-2-benzothiazolyl)thio]phenyl]acetyl]-L- cysteine) directly inhibited collagenase activity (IC50 = 15.4 microM). This inhibition was reversible upon dialysis. WY-46,135 also directly inhibited neutral protease activity (IC50 = 16.8 microM) but did not significantly block bacterial collagenase activity at a concentration of 80 microM. In contrast, WY-48,989 (4-[[2-(7-chloro-2-phenyl-2H-pyrazolo[4,3-c]quinolin-4- yl)ethyl]amino]benzonitrile) did not directly inhibit either collagenase (10 microM) or neutral protease (100 microM) activity. Both WY-48,989 and WY-46,135 inhibited IL-1 stimulated release of neutral proteases (IC50 = 3 microM). The activities of these compounds represents two potential approaches for the treatment of osteoarthritis. WY-46,135 combines direct metalloprotease inhibitory activity with the inhibition of IL-1 stimulated neutral protease release from articular chondrocytes while WY-48,989 selectively inhibits the IL-1 induced release of metalloproteases.
Subject(s)
Acetylcysteine/analogs & derivatives , Cartilage, Articular/enzymology , Cysteine/analogs & derivatives , Interleukin-1/pharmacology , Protease Inhibitors/pharmacology , Pyrazoles/pharmacology , Animals , Benzothiazoles , Cartilage, Articular/pathology , Cysteine/pharmacology , Endopeptidases/analysis , Endopeptidases/biosynthesis , Enzyme Induction/drug effects , Hindlimb , In Vitro Techniques , Male , Microbial Collagenase/analysis , RabbitsABSTRACT
The synthesis and in vitro antibacterial profile of a series of (Z)-(2-amino-4-thiazolyl)-[(2,3-dialkoxypropoxy)imino]acetyl derivatives of 7-aminocephalosporanic acid and 3-aminomonobactamic acid are reported.
