Recombinant human 92-kDa type IV collagenase/gelatinase from baculovirus-infected insect cells: expression, purification, and characterization.

Human 92-kDa type IV collagenase/gelatinase (MMP9) has been expressed in insect cells and secreted into the cell medium via a baculovirus expression system. The expression level of the proenzyme from Trichoplusia ni cells was estimated to be > = 300 mg/L of cell medium. The recombinant protein was purified in a single step using heparin-affinity chromatography with an overall yield of ca.

2,5-Diarylisothiazolone: novel inhibitors of cytokine-induced cartilage destruction.

A series of 2,5-diarylisothiazolones is reported that inhibit the IL-1 beta-induced breakdown of bovine nasal septum cartilage in an organ culture assay. The synthesis and preliminary SAR of these compounds are described. These compounds represent a novel, nonpeptide lead series approach to the mediation of the chronic cartilage breakdown associated with arthritic disease.

Comparative effects of selective cyclooxygenase 1 and cyclooxygenase 2 inhibitors on myeloperoxidase and 3 alpha-hydroxysteroid dehydrogenase.

The clinical efficacy of non-steroidal anti-inflammatory drugs (NSAIDs) is believed to result from the ability of these compounds to inhibit the inducible isoform of the enzyme cyclooxygenase, COX2. The gastrointestinal and renal side effects of these drugs, in contrast, are thought to relate to their ability to inhibit the constitutive isozyme, COX1. There is structural and pharmacological evidence that suggests that NSAIDs may also inhibit two unrelated enzymes, myeloperoxidase (MP) and 3 alpha-hydroxysteroid dehydrogenase (3 alpha-HSD), potentially with untoward consequences for the patient.

Metabolism resistant isothiazolone inhibitors of cartilage breakdown.

A series of 2-(arylmethyl)pyridoisothiazolones is reported that inhibit the IL-1 beta induced breakdown of bovine nasal septum cartilage in an organ culture assay. The synthesis and preliminary SAR of these compounds are described. These compounds represent a novel, non-peptide lead series approach to the mediation of the chronic cartilage breakdown associated with arthritic disease.

Mechanism of selective inhibition of the inducible isoform of prostaglandin G/H synthase.

Selective inhibition of the inducible isoform of prostaglandin G/H synthase (cyclooxygenase-2; COX2; EC 1.14.99.

Heteroaryl-fused 2-phenylisothiazolone inhibitors of cartilage breakdown.

The synthesis, biological evaluation, and structure-activity relationships of a series of N-phenyl heteroaryl-fused isothiazolones are described. These isothiazolones have been shown to exhibit potent, dose-dependent inhibition of IL-1 beta-induced breakdown of proteoglycan in a cartilage organ culture assay. This effect is likely due to inhibition of MMP activation and a consequent reduction in MMP activity following IL-1 beta stimulation.

Characterization of extracellular phospholipase A2 (PLA2) activity in fluid and peritoneal cells from casein-treated rats.

Extracellular phospholipase A2 activity (PLA2) found in the fluid and cells of the peritoneal cavity of rats injected with casein is described. PLA2 activities from both the fluid and cells require Ca2+ and have pH optima of 7. Acid-extraction increased PLA2 activity in the polymorphonuclear leukocyte (PMN) homogenates 20-fold but not the PLA2 activity in the extracellular fluid.

Phospholipase A2 (PLA2) activity in rabbit chondrocytes.

The effects of recombinant interleukin-1 beta (rIL-1 beta), recombinant tumor necrosis factor (rTNF alpha) and two growth factors, basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) on PLA2 activity and prostaglandin E2 (PGE2) release were investigated using rabbit chondrocytes. Cellular PLA2 activity increased 2-10 x above controls in the presence of 8 x 10(-12) M (5 U) rIL-1 beta or 5 x 10(-9) M rTNF alpha after 20 hr incubation. PLA2 activity remained constant with 1-50 ng/ml of either growth factor.

The effect of immunomodulating drugs on adjuvant-induced arthritis in Lewis rats.

The purpose of this study was to investigate the effects of cyclosporine A (CSA) and methotrexate (MTX) as potential immunomodulators in a nonestablished adjuvant arthritis (AA) model. Non-injected hind paw volumes were reduced when AA rats were treated for 18 days with CSA (100% at 10 mg/kg) or MTX (100% at 0.1 mg/kg).