David W. Snyder

Indole inhibitors of human nonpancreatic secretory phospholipase A2. 3. Indole-3-glyoxamides.

Phospholipases (PLAs) produce rate-limiting precursors in the biosynthesis of various types of biologically active lipids involved in inflammatory processes. Increased levels of human nonpancreatic secretory phospholipase A2 (hnps-PLA2) have been detected in several pathological conditions. An inhibitor of this enzyme could have therapeutic utility. A broad screening program was carried out to identify chemical structures which could inhibit hnps-PLA2. One of the lead compounds generated by the screening program was 5-methoxy-2-methyl-1-(phenylmethyl)-1H-indole-3-acetic acid (13a). We describe the syntheses, structure−activity relationships, and pharmacological activities of a series of indole-3-acetamides and related compounds derived from this lead. This SAR was undertaken with the aid of X-ray crystal structures of complexes between the inhibitors and hnps-PLA2 which were of great value in directing the SAR.

Porcine pancreatic phospholipase A2-induced contractions of guinea pig lung pleural strips

The contractile nature of porcine pancreatic phospholipase A2 (PLA2) was characterized on paired pleural strips obtained from guinea pig lung. PLA2 (0.003-10 U/ml) produced concentration-related contractile responses which were sensitive to various drugs. The major component of the PLA2-induced contractions was derived from products of the cyclooxygenase pathway since a cyclooxygenase inhibitor or the combination of a thromboxane synthetase inhibitor and a thromboxane receptor antagonist produced a 54-65% reduction of the contractile responses. 5-Lipoxygenase products contributed to a smaller component of the PLA2-induced responses since 5-lipoxygenase inhibitors or the combination of a leukotriene (LT) B4 receptor antagonist and an LTD4/LTE4 receptor antagonist only suppressed the maximal responses 22-32%. PLA2-induced contractile responses were nearly abolished by altering both sides of the arachidonic acid cascade simultaneously. In contrast, a PAF receptor antagonist, a histamine (H1) receptor antagonist and an acetylcholine receptor antagonist, failed to significantly reduce PLA2-induced responses. These results demonstrate that exogenous administration of porcine pancreatic PLA2 produced concentration-dependent contractions of pleural strips mediated through the generation of eicosanoids.

Dibasic benzo[b]thiophene derivatives as a novel class of active site directed thrombin inhibitors: 2. Sidechain optimization and demonstration of in vivo efficacy.

Potent, subnanomolar thrombin inhibitors 4, 5, and 6 are developed through side chain optimization of novel, benzo[b]thiophene-based small organic entities 2 and 3 and through SAR additivity studies of the new structural elements identified. X-ray crystallographic studies of 4b-thrombin complex revealed a hydrophobic and an electrostatic interaction of these new elements with thrombin at the S2 and S3 binding sites. In vitro and in vivo pharmacological studies showed that 4, 5, and 6 are potent anticoagulants in human plasma with demonstrated antithrombotic efficacy in a rat model of thrombosis.

Endogenously formed leukotriene C4 activates LTC4 receptors in guinea pig tracheal strips

The bioconversion of leukotriene (LT) C4 to LTD4 via gamma-glutamyl transpeptidase is clearly defined in guinea pig trachea. Acivicin, an inhibitor of gamma-glutamyl transpeptidase, was used to study the contractile responses elicited by either endogenously released or exogenously administered LTC4 and the antagonistic nature of LY 171883 and ICI 204,219, LTD4/LTE4 receptor antagonists, on guinea pig tracheal strips. Pretreating tracheal strips with acivicin resulted in a concentration-related, selective leftward shift in the LTC4 concentration-response curves. Potency of LTC4 was increased 3-fold. Likewise, antigen concentration response curves were potentiated in acivicin-pretreated tissues. Antagonism of LTC4 and antigen contractile responses by LY 171883 and ICI 204,219 were reduced or abolished by acivicin-pretreatment. In contrast, these receptor antagonists effectively blocked LTD4 responses in control and acivicin-pretreated tissues. The results demonstrated that inhibition of gamma-glutamyl transpeptidase by acivicin blocked the bioconversion of LTC4 to LTD4 regardless of the source of LTC4. Data indicated that endogenously formed LTC4 was able to activate the LTC4 receptor in guinea pig tracheal strips.

Relative potencies of 5-lipoxygenase inhibitors on antigeninduced contractions of guinea pig tracheal strips

A quantitative method to assess relative potencies (IC50) of 5-lipoxygenase (5-LO) enzyme inhibitors was established in antigen-induced contractions of tracheas isolated from actively sensitized guinea pigs (Schultz-Dale model). The relative potencies of four purported 5-LO inhibitors determined in this tissue assay were compared with those from a crude enzyme preparation isolated from guinea pig neutrophils. All compounds suppressed ovalbumin (OA)-induced tracheal contractions in a concentration-related manner in the presence of indomethacin and pyrilamine. IC50 Values, determined from the percent inhibition values obtained from responses at 30 ng/mL OA of these compounds ranged from 0.56-15 microM. A similar rank order of potency for inhibition of 5-HETE formation from a crude enzyme preparation was observed. This suggested that these agents had a common mechanism of action in the two assay systems and further validated the IC50 values determined in trachea assay. LY171883, an LTD4/LTE4 receptor antagonist, also suppressed OA-induced contractions concentration dependently with an IC50 of 4.9 microM determined by this method. LTD4 concentration-response curves were not altered by any of the four 5-LO inhibitors, ruling out the possibility that these agents were acting as LT receptor antagonists. Results of this study demonstrated that relative potencies of 5-LO inhibitors can be quantitatively assessed using this airway tissue model, which helps in identifying potential therapeutic agents for asthma.