title: Pharmacokinetics of Rac inhibitor EHop-016 in mice by ultra-performance liquid chromatography tandem mass spectrometry.
abstract: The Rho GTPase Rac is an important regulator of cancer cell migration and invasion; processes required for metastatic progression. We previously characterized the small molecule EHop-016 as a novel Rac inhibitor in metastatic breast cancer cells and recently found that EHop-016 was effective at reducing tumor growth in nude mice at 25 mg/kg bodyweight (BW). The purpose of this study was to compare the pharmacokinetics and bioavailability of EHop-016 at different dosages in a single dose input scheme (10, 20 and 40 mg/kg BW) following intraperitoneal (IP) and oral gavage (PO) administration to nude mice. We developed and validated a rapid and sensitive method for the quantitation of EHop-016 in mouse plasma by ultra high performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry (UPLC/MS/MS). Separation was carried out on an Agilent Poroshell 120 EC-C18 column (3.0 mm × 50 mm) using organic and aqueous mobile phases. EHop-016 was identified from its accurate mass and retention time from the acquired full-scan chromatogram and quantified by its peak area. The validated method was linear (R(2)>0.995) over the range of 5-1000 ng/mL (1/x(2) weighting). Pharmacokinetic parameters were obtained by non-compartmental analysis using WinNonlin. The area under the curve (AUC₀-∞) ranged from 328 to 1869 ng h/mL and 133-487 ng h/mL for IP and PO dosing, respectively. The elimination half-life (t₁/₂) ranged from 3.8-5.7 h to 3.4-26.8 h for IP and PO dosing, respectively. For both IP and PO administration, the AUC₀-∞values were proportional to the tested doses demonstrating linear PK profiles. The relative bioavailability of EHop-016 after oral gavage administration ranged from 26% to 40%. These results support further preclinical evaluation of EHop-016 as a new anti-cancer therapy.
authors.affiliation: Department of Biochemistry, School of Medicine, University of Puerto Rico, San Juan, PR, United States.
authors.forename: Tessa
authors.initials: T
authors.lastname: Humphries-Bickley
authors.affiliation: Department of Biochemistry, School of Medicine, University of Puerto Rico, San Juan, PR, United States; Department of Pathology and Laboratory Medicine, Universidad Central del Caribe, Bayamon, PR, United States.
authors.forename: Linette
authors.initials: L
authors.lastname: Castillo-Pichardo
authors.affiliation: Fundacion de Investigacion, Rio Piedras, PR, United States.
authors.forename: Francheska
authors.initials: F
authors.lastname: Corujo-Carro
authors.affiliation: Department of Pharmaceutical Sciences, School of Pharmacy, University of Puerto Rico, San Juan, PR, United States.
authors.forename: Jorge
authors.initials: J
authors.lastname: Duconge
authors.affiliation: Department of Pharmaceutical Sciences, School of Pharmacy, University of Puerto Rico, San Juan, PR, United States.
authors.forename: Eliud
authors.initials: E
authors.lastname: Hernandez-O'Farrill
authors.affiliation: Department of Pharmaceutical Sciences, School of Pharmacy, University of Puerto Rico, San Juan, PR, United States.
authors.forename: Cornelis
authors.initials: C
authors.lastname: Vlaar
authors.affiliation: Department of Biochemistry, School of Medicine, University of Puerto Rico, San Juan, PR, United States; Fundacion de Investigacion, Rio Piedras, PR, United States.
authors.forename: Jose F
authors.initials: JF
authors.lastname: Rodriguez-Orengo
authors.affiliation: Department of Anatomy and Cell Biology, Universidad Central del Caribe, Bayamon, PR, United States.
authors.forename: Luis
authors.initials: L
authors.lastname: Cubano
authors.affiliation: Department of Biochemistry, School of Medicine, University of Puerto Rico, San Juan, PR, United States. Electronic address: su.d@upr.edu.
authors.forename: Suranganie
authors.initials: S
authors.lastname: Dharmawardhane
mesh_terms: D000818:Animals; D002227:Carbazoles / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005260:Female; D016014:Linear Models; D051379:Mice; D008819:Mice, Nude; D011743:Pyrimidines / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*; D020744:rac GTP-Binding Proteins / Q000037:antagonists & inhibitors*
chemical_list: D002227:Carbazoles; C572775:N4-(9-ethyl-9H-carbazol-3-yl)-N2-(3-morpholin-4-ylpropyl)-pyrimidine-2,4-diamine; D011743:Pyrimidines; D020744:rac GTP-Binding Proteins
keywords: Cancer; EHop-016; Pharmacokinetics; Rac; UPLC/MS/MS