Liquid chromatography-tandem mass spectrometry based method development and validation of S016-1271 (LR8P), a novel cationic antimicrobial peptide for its application to pharmacokinetic studies.

Autor: Puttrevu SK; Pharmacokinetics and Metabolism Division, CSIR-Central Drug Research Institute, Lucknow, 226031, India; Academy of Scientific and Innovative Research, New Delhi, 110001, India., Laxman TS; Pharmacokinetics and Metabolism Division, CSIR-Central Drug Research Institute, Lucknow, 226031, India; Academy of Scientific and Innovative Research, New Delhi, 110001, India., Tripathi AK; Molecular and Structural Biology Division, CSIR-Central Drug Research Institute, Lucknow, 226031, India; Jawaharlal Nehru University, New Delhi, 110067, India., Yadav AK; Pharmacokinetics and Metabolism Division, CSIR-Central Drug Research Institute, Lucknow, 226031, India., Verma SK; Pharmacokinetics and Metabolism Division, CSIR-Central Drug Research Institute, Lucknow, 226031, India; Jawaharlal Nehru University, New Delhi, 110067, India., Mishra A; Pharmacokinetics and Metabolism Division, CSIR-Central Drug Research Institute, Lucknow, 226031, India; Academy of Scientific and Innovative Research, New Delhi, 110001, India., Pradhan R; Pharmacokinetics and Metabolism Division, CSIR-Central Drug Research Institute, Lucknow, 226031, India; National Institute of Pharmaceutical Education and Research, Raebareli, 229010, India., Verma NK; Molecular and Structural Biology Division, CSIR-Central Drug Research Institute, Lucknow, 226031, India; Jawaharlal Nehru University, New Delhi, 110067, India., Ghosh JK; Academy of Scientific and Innovative Research, New Delhi, 110001, India; Molecular and Structural Biology Division, CSIR-Central Drug Research Institute, Lucknow, 226031, India. Electronic address: jk_ghosh@cdri.res.in., Bhatta RS; Pharmacokinetics and Metabolism Division, CSIR-Central Drug Research Institute, Lucknow, 226031, India; Academy of Scientific and Innovative Research, New Delhi, 110001, India. Electronic address: rabi_bhatta@cdri.res.in.
Jazyk: angličtina
Zdroj: Journal of pharmaceutical and biomedical analysis [J Pharm Biomed Anal] 2019 May 30; Vol. 169, pp. 116-126. Date of Electronic Publication: 2019 Feb 19.
DOI: 10.1016/j.jpba.2019.01.046
Abstrakt: S016-1271 (LR8P) is a broad spectrum novel cationic antimicrobial peptide. The objective of the present study was to develop a selective liquid chromatography-tandem mass spectrometry (LC-MS/MS) based bioanalytical method of S016-1271 peptide in mice and human plasma in order to uncover its pharmacokinetic aspects. The chromatographic separation of S016-1271 (FR8P as internal standard) was achieved on a Waters™ X select CSH-C18 column (75 × 3.0 mm, 2.5 μ) using mixture of acetonitrile and triple distilled water (TDW) both containing 0.05% formic acid as mobile phase. A seven minute linear gradient method was designed to separate analytes from ion suppression at a flow rate of 0.3 mL/min. The extraction of analytes from mice and human plasma was performed through solid phase extraction technique using mixed mode weak cation exchange cartridge (Thermo SOLA WCX 10 mg 1CC) with an extraction recovery of analytes about 75%. Mass spectrometric detection of S016-1271 and FR8P was performed with optimized multiple reaction monitoring (MRM) transitions (Q1/Q3) at 658.8 [M+3H] 3+ /653.2 [M+3H-NH 3 ] 3+ and 443.4 [M+5H] 5+ /434.7 [y 12 -NH 3 ] 4+ ,respectively in positive electrospray ionization (ESI) mode. The linearity in mice and human plasma was established over a concentration range of 7.81-250 ng/mL with regression coefficient (r 2  > 0.99). The currently developed method was validated as per US-FDA guidelines and found to be within the acceptable limits. The method was successfully applied to intravenous (IV) pharmacokinetic study in mice wherein the levels were detected upto 24 h. The peptide demonstrated poor distribution characteristics which were demonstrated through volume of distribution at steady state (202.71 ± 47.02 mL/kg less than total body water of mice; 580 mL/kg). The clearance of the peptide predominantly occurred through central compartment (central clearance is 25 fold greater than peripheral clearance). Also, the in vitro pharmacokinetic studies demonstrated the stability of S016-1271 in plasma and high plasma protein binding in mice and humans.
(Copyright © 2019. Published by Elsevier B.V.)
Databáze: MEDLINE
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