Enhanced responsiveness of platelets to vicagrel in IL‐10‐deficient mice through STAT3‐dependent up‐regulation of the hydrolase arylacetamide deacetylase in the intestine

Abstract

Background and Purpose: Vicagrel is a novel promising antiplatelet drug designed for overcoming clopidogrel resistance. There is limited evidence indicating that exogenous IL-10 suppresses CYP3A4 activity in healthy subjects and that IL-10 knockout (KO) mice exhibit increased clopidogrel bioactivation compared with wild-type (WT) mice. In this study, we sought to determine whether IL-10 could play an important role in the metabolism of and platelet response to vicagrel in mice. Experimental Approach: IL-10 KO and WT mice were administered vicagrel, then their plasma H4 (active metabolite of vicagrel) concentrations were determined by LC-MS/MS, and inhibition of ADP-induced whole-blood platelet aggregation by vicagrel was assessed with an aggregometer. The mRNA and protein levels of several relevant genes between IL-10 KO and WT mice were measured by qRT-PCR and Western blots, respectively. Intestinal Aadac protein levels were measured in IL-10 WT mice injected i.p. with vehicle control, Stattic, or BAY 11-7082. Key Results: Compared with WT mice, IL-10 KO mice exhibited significantly increased plasma levels of H4 and enhanced platelet responses to vicagrel, as well as significantly higher mRNA and protein levels of arylacetamide deacetylase (Aadac) in the intestine. In WT mice, STAT3, not NF-κB, mediated Aadac expression in the intestine. Conclusions and Implications: IL-10 suppresses metabolic activation of vicagrel through down-regulation of Aadac in mouse intestine in a STAT3-dependent manner and, consequently, attenuates platelet responses to vicagrel, suggesting that the antiplatelet effect of vicagrel may be modulated by changes in plasma IL-10 levels in relevant clinical settings. 1 | INTRODUCTION A large number of relevant studies have demonstrated that inflammatory cytokines are involved in the whole process of atherosclerosis, due to their excessive generation and uninterrupted release in response to inflammatory stimuli (Ramji & Davies, 2015), and that inflammation plays a critical role in the pathogenesis and progression

Impacts of CYP2C19 genetic polymorphisms on bioavailability and effect on platelet adhesion of vicagrel, a novel thienopyridine P2Y12 inhibitor

“…Vicagrel and clopidogrel have pharmacological effects based on the same active thiol metabolite, H4, which is formed through a 2‐step metabolic activation process (Figure 1). 12,23–27 The second step, from 2‐oxo‐clopidogrel to the thiol metabolite, H4, is the same for both vicagrel and clopidogrel. However, for vicagrel, the first step is the rapid and complete hydrolysis of its ester function to form 2‐oxo‐clopidogrel, which is mediated by human carboxylesterase 2 and arylacetamide deacetylase in the gut instead of CYP2C19 in the liver.…”

Role of circadian clock in the chronoefficacy and chronotoxicity of clopidogrel

“…Whole‐blood samples (400 μl) were collected into a heparinized tube via cardiac puncture. Whole‐blood platelet aggregation induced by ADP (10 μM, final concentration) was measured using the whole‐blood aggregometer as previously described (Jia et al, 2019). The inhibition percentage of clopidogrel in ADP‐induced platelet aggregation was calculated as [1 − (value of a treated mouse/value of a control mouse)] × 100%.…”

Fluorine impairs carboxylesterase 1-mediated hydrolysis of T-2 toxin and increases its chondrocyte toxicity

“…To determine whether NaF could modulate the expression level of CES1, we extracted total RNA from HepG2 cells using Trizol reagent (Invitrogen, Carlsbad, California, USA) according to the manufacturer's instructions and were performed as described elsewhere ( 26 , 27 ). The following primer sequences were used: for CES1 : 5′- AGTTTCAGTACCGTCCAAGC-3′ (forward) and 5′- CCATCTTGCTAAGTCTGATCTCC-3′ (reverse); for GAPDH : 5′- GCACCGTCAAGGCTGAGAAC-3′ (forward), and 5′- TGGTGAAGACGCCAGTGGA-3′ (reverse).…”