The parmodulin NRD-21 is an allosteric inhibitor of PAR1 Gq signaling with improved anti-inflammatory activity and stability

Gandhi, Disha M.; Rosas, Ricardo; Greve, Eric; Kentala, Kaitlin; Diby, N'Guessan; Snyder, Vladyslava; Stephans, Allison; Yeung, Teresa H.W.; Subramaniam, Saravanan; DiMilo, Elliot; Kurtenbach, Khia E.; Arnold, Leggy A.; Weiler, Hartmut; Dockendorff, Chris

doi:10.1016/j.bmc.2019.06.043

Cited by 10 publications

(5 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As a result, oxazole‐based derivative NRD‐21 (Fig. 6, IC 50 0.37 ± 0.13 μ m ) was identified as a negative allosteric PAR1 selective and reversible modulator with improved plasma stability [235]. SAR studies on NRD‐21 determined that limited size lipophilic groups substituted in the 1,3‐diaminobenzene scaffold of ML161 enhance the inhibition of PAR1‐induced Gα q signaling [235].…”

Section: Synthetic Pars Modulators and Structure–activity Relationshipmentioning

confidence: 99%

Molecular mechanisms regulating Proteinase‐Activated Receptors (PARs)

Chandrabalan

Ramachandran

2021

The FEBS Journal

View full text Add to dashboard Cite

Proteinase activated receptors (PARs) are a four-member family of G protein-coupled receptors defined by their irreversible proteolytic mechanism of activation. PARs have emerged as important regulators of various physiological responses and are implicated in numerous pathological conditions. Importantly, PAR1 and PAR4 are critical regulators of platelet function, while PAR2 is well established as a driver of inflammatory responses. PAR targeted drug development efforts are therefore of great interest. In this review, we provide an overview of recent advances in our understanding of molecular mechanisms underlying PAR activation, effector interaction and signalling. We also provide an overview of the diverse proteolytic enzymes that are now established as PAR regulators and describe the ability of different enzymes to elicit biased signalling through PARs. Finally, we highlight recent advances in the development of PAR targeted pharmacological agents and discuss recent structure-activity relationship studies.

show abstract

Section: Synthetic Pars Modulators and Structure–activity Relationshipmentioning

confidence: 99%

Molecular mechanisms regulating Proteinase‐Activated Receptors (PARs)

Chandrabalan

Ramachandran

2021

The FEBS Journal

View full text Add to dashboard Cite

show abstract

“…H1-R blocking using CET had no effects, indicating that this receptor is not involved in DAM mediated enhanced endothelial permeability. ML161 acts at the intracellular side of PAR-1 and selectively blocks G q 22 mediated signal transduction via Ca 2+ /Calmodulin towards eNOS signaling 23 . ROCK inhibition blocks the signal transduction towards myosin light chain kinase phosphorylation and actomyosin contraction 24 .…”

Section: Discussionmentioning

confidence: 99%

Amphetamine increases vascular permeability by modulating endothelial actin cytoskeleton and NO synthase via PAR-1 and VEGF-R

Böttner,

Fischer-Schaepmann,

Werner

et al. 2024

Sci Rep

View full text Add to dashboard Cite

Abuse of amphetamine-type stimulants is linked to cardiovascular adverse effects like arrhythmias, accelerated atherosclerosis, acute coronary syndromes and sudden cardiac death. Excessive catecholamine release following amphetamine use causes vasoconstriction and vasospasms, over time leading to hypertension, endothelial dysfunction or even cardiotoxicity. However, immediate vascular pathomechanisms related to amphetamine exposure, especially endothelial function, remain incompletely understood and were analyzed in this study. Pharmaco-pathological effects of acute d-amphetamine-sulfate (DAM) were investigated ex vivo using contraction–force measurements of rat carotid artery rings and in vitro using label-free, real-time electrochemical impedance spectroscopy (EIS) on endothelial and smooth muscle cells. Specific receptor and target blocking was used to identify molecular targets and to characterize intracellular signaling. DAM induced vasodilation represented by 29.3±2.5% decrease in vascular tone (p<0.001) involving vascular endothelial growth factor receptor (VEGF-R) and protease activated receptor 1 (PAR-1). EIS revealed that DAM induces endothelial barrier disruption (−75.9±1.1% of initial cellular impedance, p<0.001) also involving VEGF-R and PAR-1. Further, in response to DAM, Rho-associated protein kinase (ROCK) mediated reversible contraction of actin cytoskeleton resulting in endothelial barrier disruption. Dephosphorylation of Serine1177 (−50.8±3.7%, p<0.001) and Threonine495 (−44.8±6.5%, p=0.0103) of the endothelial NO synthase (eNOS) were also observed. Blocking of VEGF-R and PAR-1 restored baseline eNOS Threonine495 phosphorylation. DAM induced vasodilation, enhanced vascular permeability and actin cytoskeleton contraction and induced eNOS hypophosphorylation involving VEGF-R, PAR-1 and ROCK. These results may contribute to a better understanding of severe adverse cardiovascular effects in amphetamine abuse.

show abstract

“…Moreover, Peptidase M84 failed to induce apoptosis in PAR1 silenced ovarian cancer cells. Furthermore, ML161, a PAR1 mediated Gq signalling inhibitor [81] also showed a significant decrease in apoptosis in Peptidase M84 treated cells. This suggests that Peptidase M84 caused PAR-1 activation through Gq signalling induction.…”

Section: Discussionmentioning

confidence: 99%

Metallo-protease Peptidase M84 fromBacillus altitudinisinduces ROS dependent apoptosis in ovarian cancer cells by targeting PAR-1

Nag,

Ray,

Tapader

et al. 2023

Preprint

View full text Add to dashboard Cite

In pursuit of isolating novel anticancer proteases from environmental microbial isolates, we have purified and identified an extracellular metallo-protease from Bacillus altitudinis named Peptidase M84. This protease selectively triggered apoptosis in human ovarian adenocarcinoma cells (PA-1, SKOV3) and mouse ovarian carcinoma cells (ID8), in addition to exhibiting no significant effect on normal human epithelial ovarian cell (IOSE) and mouse peritoneal macrophage (PEMΦ) cell viabilities. Protease activated receptor-1 (PAR-1); a GPCR which is reported to be overexpressed in ovarian cancer cells was identified as a novel target of Peptidase M84. We observed that Peptidase M84 induced PAR-1 overexpression along with activating its downstream signalling effectors NFκB and MAPK to promote excessive reactive oxygen species (ROS) generation in ovarian cancer cells. This disrupted mitochondrial membrane potential, allowed cytosolic release of mitochondrial cytochrome c, increased the Bax (pro-apoptotic) to Bcl-2 (anti-apoptotic) ratio and promoted DNA damage to evoke apoptotic death of the ovarian cancer cells. Peptidase M84 also reduced nuclear ki-67 expression in these malignant cells to render an anti-proliferative role. In in vivo set-up, weekly intraperitoneal administration of Peptidase M84 (12 ug/kg body-weight) in the ID8 mice model significantly diminished ascitic fluid accumulation through induction of oxidative stress, increasing murine survival rates by 60%. Collectively, our in vitro and in vivo findings suggested that Peptidase M84 triggered PAR-1 mediated oxidative stress to act as an apoptosis inducer in ovarian cancer cells. This established Peptidase M84 as a promising drug candidate for receptor mediated targeted-therapy of ovarian cancer.

show abstract

The parmodulin NRD-21 is an allosteric inhibitor of PAR1 Gq signaling with improved anti-inflammatory activity and stability

Cited by 10 publications

References 32 publications

Molecular mechanisms regulating Proteinase‐Activated Receptors (PARs)

Molecular mechanisms regulating Proteinase‐Activated Receptors (PARs)

Amphetamine increases vascular permeability by modulating endothelial actin cytoskeleton and NO synthase via PAR-1 and VEGF-R

Metallo-protease Peptidase M84 fromBacillus altitudinisinduces ROS dependent apoptosis in ovarian cancer cells by targeting PAR-1

Contact Info

Product

Resources

About