In vitro pharmacokinetic behavior in lung of harringtonine, an antagonist of SARS-CoV-2 associated proteins: New insights of inhalation therapy for COVID-19

Jiapan Gao; Panpan Lei; Xinyue Su; Jinna Liang; Bingxi Ren; Xiaoyu Ma; Yuxiu Zhang; Yongjing Zhang; Weina Ma

doi:10.1016/j.phymed.2024.155582

In vitro pharmacokinetic behavior in lung of harringtonine, an antagonist of SARS-CoV-2 associated proteins: New insights of inhalation therapy for COVID-19

Phytomedicine. 2024 Apr 4:129:155582. doi: 10.1016/j.phymed.2024.155582. Online ahead of print.

Authors

Jiapan Gao¹, Panpan Lei¹, Xinyue Su¹, Jinna Liang¹, Bingxi Ren¹, Xiaoyu Ma¹, Yuxiu Zhang¹, Yongjing Zhang², Weina Ma³

Affiliations

¹ School of Pharmacy, Xi'an Jiaotong University, Xi'an, 710061, China.
² School of Pharmacy, Xi'an Jiaotong University, Xi'an, 710061, China; Department of Medical Oncology, First Affiliated Hospital of Xi'an Jiaotong University, Xi'an Jiaotong University, Xi'an, 710061, China.
³ School of Pharmacy, Xi'an Jiaotong University, Xi'an, 710061, China. Electronic address: maweina2015@xjtu.edu.cn.

PMID: 38608595
DOI: 10.1016/j.phymed.2024.155582

Abstract

Background: Recent studies have shown that harringtonine (HT) could specifically bind with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike (S) protein and host cell transmembrane serine protease 2 (TMPRSS2) to block membrane fusion, which is an effective antagonist for SARS-CoV-2.

Purpose: Our study focused on in-depth exploration of in vitro pharmacokinetic characteristics of HT in lung.

Methods: HPLC-fluorescence detection method was used to detect changes of HT content. Incubation systems of lung microsomes for phase I metabolism and UGT incubation systems for phase II metabolism were performed to elucidate metabolites and metabolic mechanisms of HT, and then the metabolic enzyme phenotypes for HT were clarified by chemical inhibition method and recombinant enzyme method. Through metabolomics, we comprehensively evaluated the physiological dynamic changes in SD rat and human lung microsomes, and revealed the relationship between metabolomics and pharmacological activity of HT.

Results: HPLC-fluorescence detection method showed strong specificity, high accuracy, and good stability for rapid quantification of HT. We confirmed that HT mainly underwent phase I metabolism, and the metabolites of HT in different species were all identified as 4'-demethyl HT, with metabolic pathway being hydrolysis reaction. CYP1A2 and CYP2E1 participated in HT metabolism, but as HT metabolism was not NADPH dependent, the esterase HCES1 in lung also played a role. The main KEGG pathways in SD rat and human lung microsomes were cortisol synthesis and secretion, steroid hormone biosynthesis and linoleic acid metabolism, respectively. The downregulated key biomarkers of 11-deoxycortisol, 21-deoxycortisol and 9(10)-EpOME suggested that HT could prevent immunosuppression and interfere with infection and replication of SARS-CoV-2.

Conclusion: HT was mainly metabolized into 4'-demethyl HT through phase I reactions, which was mediated by CYP1A2, CYP2E1, and HCES1. The downregulation of 11-deoxycortisol, 21-deoxycortisol and 9(10)-EpOME were key ways of HT against SARS-CoV-2. Our study was of great significance for development and clinical application of HT in the treatment of COVID-19.

Keywords: HPLC-fluorescence; Harringtonine; Lung microsomes; Metabolomics; SARS-CoV-2.