Discovery of a Novel Potent and Selective Calcium Release-Activated Calcium Channel Inhibitor: 2,6-Difluoro-N-(2'-methyl-3'-(4-methyl-5-oxo-4,5-dihydro-1,3,4-oxadiazol-2-yl)-[1,1'-biphenyl]-4-yl)benzamide. Structure-Activity Relationship and Preclinical Characterization.

The role of calcium release-activated calcium (CRAC) channels is well characterized and is of particular importance in T-cell function. CRAC channels are involved in the pathogenesis of several autoimmune diseases, making it an attractive therapeutic target for treating inflammatory diseases, like rheumatoid arthritis (RA). A systematic structure-activity relationship study with the goal of optimizing lipophilicity successfully yielded two lead compounds, 36 and 37. Both compounds showed decent potency and selectivity and a remarkable pharmacokinetic profile. Further characterization in in vivo RA models and subsequent histopathological evaluation of tissues led to the identification of 36 as a clinical candidate. Compound 36 displayed an excellent safety profile and had a sufficient safety margin to qualify it for use in human testing. Oral administration of 36 in Phase 1 clinical study in healthy volunteers established favorable safety, tolerability, and good target engagement as measured by levels of IL-2 and TNF-α.

A novel fibrinolytic enzyme from marine Pseudomonas aeruginosa KU1 and its rapid in vivo thrombolysis with little haemolysis.

A direct acting, extracellular, fibrinolytic enzyme, ~50 KDa from marine Pseudomonas aeruginosa KU1 (PEKU1), was purified. It was found to be a metalloprotease. 60% of the total activity of the purified PEKU1 was retained at 70 °C and the enzyme was practically denatured at 80 °C, 2 h. Metal ions, such as Na+, K+ and Co2+, were found to enhance slightly the fibrinolytic activity, while Fe2+, Mn2+ and Zn2+ were found to be inhibiting. The enzyme showed only less than 5% haemolysis, suggesting its thrombolytic administration safe. Tryptic digestion revealed its similarity to serralysin like alkaline protease of P. aeruginosa. In silico studies showed its binding of protease substrates and fibrin D-dimer in its active site. High affinity binding of bradykinin to the active site of PEKU1, confirmed by in vitro cleaving, suggested its future use as an analgesic. The purified enzyme with Na+, K+ and Co2+, and without Fe2+, Mn2+ and Zn2+ showed thrombolysis in vivo in carrageenan induced murine tail thrombolytic model. The enzyme PEKU1, a novel protease from marine isolate Pseudomonas aeruginosa KU1 has great potential to be developed as a therapeutic agent to combat cardiovascular diseases, as well as analgesic and anti-inflammatory drug in appropriate sites.

Fibrinolytic Enzymes for Thrombolytic Therapy.

Cardiovascular diseases are a group of disorders consisting importantly of coronary heart disease, peripheral arterial disease, cerebrovascular disease, rheumatic heart disease, congenital heart disease, deep vein thrombosis and pulmonary embolism. Severe cardiovascular disease conditions lead to acute myocardial infarction and stroke. One of the reasons for this is formation of blood clots inside the vessel. Anticoagulants and antiplatelet drugs are used for managing cardiovascular diseases for a long time. However, they were unable to dissolve an existing thrombus. Fibrinolytic enzymes have become more substantial for treating cardiovascular diseases since they could lyse the fibrin clot within the blood vessel. Inability of plasma fibrinolytic system demands better thrombolytic drugs. Major thrombolytic enzymes belonging to plasminogen activators and plasmin like enzymes. Currently used fibrinolytic enzymes and their limitations are revisited in the present chapter. Reported enzymes from various sources with potential to be used as cardiovascular therapeutic is also discussed here.

Canine filarial infections in a human Brugia malayi endemic area of India.

A very high prevalence of microfilaremia of 42.68 per cent out of 164 canine blood samples examined was observed in Cherthala (of Alappuzha district of Kerala state), a known human Brugia malayi endemic area of south India. The species of canine microfilariae were identified as Dirofilaria repens, Brugia malayi, and Acanthocheilonema reconditum. D. repens was the most commonly detected species followed by B. pahangi. D. immitis was not detected in any of the samples examined. Based on molecular techniques, microfilariae with histochemical staining pattern of "local staining at anal pore and diffuse staining at central body" was identified as D. repens in addition to those showing acid phosphatase activity only at the anal pore. Even though B. malayi like acid phosphatase activity was observed in few dogs examined, they were identified as genetically closer to B. pahangi. Hence, the possibility of dogs acting as reservoirs of human B. malayi in this area was ruled out.

An Assessment of Antibacterial Activity of Four Endodontic Sealers on Enterococcus faecalis by a Direct Contact Test: An In Vitro Study.

Aim. To evaluate the antibacterial activity of four endodontic sealers on Enterococcus faecalis by a direct contact test. Material and Methods. Enterococcus faecalis was used as a test organism. Direct contact test which is based on measuring the effect of close contact between test bacteria and tested material on the kinetics of bacterial growth was performed to overcome the disadvantages of agar diffusion test. The sealers tested were zinc oxide eugenol-based sealer, glass-ionomer-based sealer, polydimethyl-siloxane-based sealer, and urethane dimethacrylate resin-based sealer. Data was collected by recording the optical density with the help of a spectrophotometer. Results. The sealers exhibited different inhibitory effects. The results obtained were subjected to statistical analysis by Kruskal Wallis analysis of variance and Dunn's multiple comparison test. Group comparison showed very highly significant difference between the groups. Conclusion. Zinc oxide eugenol-based sealer was the most effective and urethane dimethacrylate resin-based sealer was the least effective against Enterococcus faecalis, whereas glass-ionomer-based and polydimethyl-siloxane-based sealers were effective only for a short period. Inhibition of the bacterial growth is related to the direct contact of the microorganism with the sealer.