PMID- 35779861 OWN - NLM STAT- MEDLINE DCOM- 20220818 LR - 20240216 IS - 1521-0103 (Electronic) IS - 0022-3565 (Print) IS - 0022-3565 (Linking) VI - 382 IP - 3 DP - 2022 Sep TI - Physiological Characterization of the Transporter-Mediated Uptake of the Reversible Male Contraceptive H2-Gamendazole Across the Blood-Testis Barrier. PG - 299-312 LID - 10.1124/jpet.122.001195 [doi] AB - The blood-testis barrier (BTB) is formed by a tight network of Sertoli cells (SCs) to limit the movement of reproductive toxicants from the blood into the male genital tract. Transporters expressed at the basal membranes of SCs also influence the disposition of drugs across the BTB. The reversible, nonhormonal contraceptive, H2-gamendazole (H2-GMZ), is an indazole carboxylic acid analog that accumulates over 10 times more in the testes compared with other organs. However, the mechanism(s) by which H2-GMZ circumvents the BTB are unknown. This study describes the physiologic characteristics of the carrier-mediated process(es) that permit H2-GMZ and other analogs to penetrate SCs. Uptake studies were performed using an immortalized human SC line (hT-SerC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Uptake of H2-GMZ and four analogs followed Michaelis-Menten transport kinetics (one analog exhibited poor penetration). H2-GMZ uptake was strongly inhibited by indomethacin, diclofenac, MK-571, and several analogs. Moreover, H2-GMZ uptake was stimulated by an acidic extracellular pH, reduced at basic pHs, and independent of extracellular Na(+), K(+), or Cl(-) levels, which are intrinsic characteristics of OATP-mediated transport. Therefore, the characteristics of H2-GMZ transport suggest that one or more OATPs may be involved. However, endogenous transporter expression in wild-type Chinese hamster ovary (CHO), Madin-Darby canine kidney (MDCK), and human embryonic kidney-293 (HEK-293) cells limited the utility of heterologous transporter expression to identify a specific OATP transporter. Altogether, characterization of the transporters involved in the flux of H2-GMZ provides insight into the selectivity of drug disposition across the human BTB to understand and overcome the pharmacokinetic and pharmacodynamic difficulties presented by this barrier. SIGNIFICANCE STATEMENT: Despite major advancements in female contraceptives, male alternatives, including vasectomy, condom usage, and physical withdrawal, are antiquated and the widespread availability of nonhormonal, reversible chemical contraceptives is nonexistent. Indazole carboxylic acid analogs such as H2-GMZ are promising new reversible, antispermatogenic drugs that are highly effective in rodents. This study characterizes the carrier-mediated processes that permit H2-GMZ and other drugs to enter Sertoli cells and the observations made here will guide the development of drugs that effectively circumvent the BTB. CI - Copyright (c) 2022 by The American Society for Pharmacology and Experimental Therapeutics. FAU - Hau, Raymond K AU - Hau RK AD - Department of Pharmacology and Toxicology, College of Pharmacy (R.K.H., N.J.C.), and Department of Physiology, College of Medicine (S.H.W.), The University of Arizona, Tucson, Arizona; Department of Molecular and Integrative Physiology, KU School of Medicine, The University of Kansas Medical Center, Kansas City, Kansas (J.S.T.); Department of Medicinal Chemistry and Institute for Therapeutics Discovery and Development, College of Pharmacy, The University of Minnesota, Minneapolis, Minnesota (G.I.G.). FAU - Tash, Joseph S AU - Tash JS AD - Department of Pharmacology and Toxicology, College of Pharmacy (R.K.H., N.J.C.), and Department of Physiology, College of Medicine (S.H.W.), The University of Arizona, Tucson, Arizona; Department of Molecular and Integrative Physiology, KU School of Medicine, The University of Kansas Medical Center, Kansas City, Kansas (J.S.T.); Department of Medicinal Chemistry and Institute for Therapeutics Discovery and Development, College of Pharmacy, The University of Minnesota, Minneapolis, Minnesota (G.I.G.). FAU - Georg, Gunda I AU - Georg GI AD - Department of Pharmacology and Toxicology, College of Pharmacy (R.K.H., N.J.C.), and Department of Physiology, College of Medicine (S.H.W.), The University of Arizona, Tucson, Arizona; Department of Molecular and Integrative Physiology, KU School of Medicine, The University of Kansas Medical Center, Kansas City, Kansas (J.S.T.); Department of Medicinal Chemistry and Institute for Therapeutics Discovery and Development, College of Pharmacy, The University of Minnesota, Minneapolis, Minnesota (G.I.G.). FAU - Wright, Stephen H AU - Wright SH AD - Department of Pharmacology and Toxicology, College of Pharmacy (R.K.H., N.J.C.), and Department of Physiology, College of Medicine (S.H.W.), The University of Arizona, Tucson, Arizona; Department of Molecular and Integrative Physiology, KU School of Medicine, The University of Kansas Medical Center, Kansas City, Kansas (J.S.T.); Department of Medicinal Chemistry and Institute for Therapeutics Discovery and Development, College of Pharmacy, The University of Minnesota, Minneapolis, Minnesota (G.I.G.). FAU - Cherrington, Nathan J AU - Cherrington NJ AD - Department of Pharmacology and Toxicology, College of Pharmacy (R.K.H., N.J.C.), and Department of Physiology, College of Medicine (S.H.W.), The University of Arizona, Tucson, Arizona; Department of Molecular and Integrative Physiology, KU School of Medicine, The University of Kansas Medical Center, Kansas City, Kansas (J.S.T.); Department of Medicinal Chemistry and Institute for Therapeutics Discovery and Development, College of Pharmacy, The University of Minnesota, Minneapolis, Minnesota (G.I.G.) cherring@pharmacy.arizona.edu. LA - eng GR - R01 ES028668/ES/NIEHS NIH HHS/United States GR - P30 CA023074/CA/NCI NIH HHS/United States GR - HHSN275201300017C/HD/NICHD NIH HHS/United States GR - T32 ES007091/ES/NIEHS NIH HHS/United States GR - R01 GM123643/GM/NIGMS NIH HHS/United States GR - HHSN275201300017I/HD/NICHD NIH HHS/United States GR - R01 GM129777/GM/NIGMS NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural DEP - 20220702 PL - United States TA - J Pharmacol Exp Ther JT - The Journal of pharmacology and experimental therapeutics JID - 0376362 RN - 0 (Carboxylic Acids) RN - 0 (Contraceptive Agents, Male) RN - 0 (Indazoles) RN - 0 (Membrane Transport Proteins) RN - 0 (Organic Anion Transporters) RN - 0 (gamendazole) SB - IM MH - Animals MH - Blood-Testis Barrier MH - CHO Cells MH - Carboxylic Acids/metabolism/pharmacology MH - Chromatography, Liquid MH - *Contraceptive Agents, Male/metabolism/pharmacology MH - Cricetinae MH - Cricetulus MH - Dogs MH - Female MH - HEK293 Cells MH - Humans MH - Indazoles/pharmacology MH - Male MH - Membrane Transport Proteins/metabolism MH - *Organic Anion Transporters/metabolism MH - Tandem Mass Spectrometry PMC - PMC9426764 EDAT- 2022/07/03 06:00 MHDA- 2022/08/19 06:00 PMCR- 2023/09/01 CRDT- 2022/07/02 20:42 PHST- 2022/03/07 00:00 [received] PHST- 2022/06/14 00:00 [accepted] PHST- 2022/07/03 06:00 [pubmed] PHST- 2022/08/19 06:00 [medline] PHST- 2022/07/02 20:42 [entrez] PHST- 2023/09/01 00:00 [pmc-release] AID - jpet.122.001195 [pii] AID - JPET_AR2022001195 [pii] AID - 10.1124/jpet.122.001195 [doi] PST - ppublish SO - J Pharmacol Exp Ther. 2022 Sep;382(3):299-312. doi: 10.1124/jpet.122.001195. Epub 2022 Jul 2.