PMID- 38245951 OWN - NLM STAT- MEDLINE DCOM- 20240219 LR - 20240219 IS - 1090-2120 (Electronic) IS - 0045-2068 (Linking) VI - 144 DP - 2024 Mar TI - In vitro safety signals for potential clinical development of the anti-inflammatory pregnane X receptor agonist FKK6. PG - 107137 LID - S0045-2068(24)00042-7 [pii] LID - 10.1016/j.bioorg.2024.107137 [doi] AB - Based on the mimicry of microbial metabolites, functionalized indoles were demonstrated as the ligands and agonists of the pregnane X receptor (PXR). The lead indole, FKK6, displayed PXR-dependent protective effects in DSS-induced colitis in mice and in vitro cytokine-treated intestinal organoid cultures. Here, we report on the initial in vitro pharmacological profiling of FKK6. FKK6-PXR interactions were characterized by hydrogen-deuterium exchange mass spectrometry. Screening FKK6 against potential cellular off-targets (G protein-coupled receptors, steroid and nuclear receptors, ion channels, and xenobiotic membrane transporters) revealed high PXR selectivity. FKK6 has poor aqueous solubility but was highly soluble in simulated gastric and intestinal fluids. A large fraction of FKK6 was bound to plasma proteins and chemically stable in plasma. The partition coefficient of FKK6 was 2.70, and FKK6 moderately partitioned into red blood cells. In Caco2 cells, FKK6 displayed high permeability (A-B: 22.8 x 10-6 cm.s(-1)) and no active efflux. These data are indicative of essentially complete in vivo absorption of FKK6. The data from human liver microsomes indicated that FKK6 is rapidly metabolized by cytochromes P450 (t(1/2) 5 min), notably by CYP3A4. Two oxidized FKK6 derivatives, including DC73 (N6-oxide) and DC97 (C19-phenol), were detected, and these metabolites had 5-7 x lower potency as PXR agonists than FKK6. This implies that despite high intestinal absorption, FKK6 is rapidly eliminated by the liver, and its PXR effects are predicted to be predominantly in the intestines. In conclusion, the PXR ligand and agonist FKK6 has a suitable pharmacological profile supporting its potential preclinical development. CI - Copyright (c) 2024 Elsevier Inc. All rights reserved. FAU - Dvorak, Zdenek AU - Dvorak Z AD - Department of Cell Biology and Genetics, Faculty of Science, Palacky University, Slechtitelu 27, 783 71 Olomouc, Czech Republic. Electronic address: zdenek.dvorak@upol.cz. FAU - Vyhlidalova, Barbora AU - Vyhlidalova B AD - Department of Cell Biology and Genetics, Faculty of Science, Palacky University, Slechtitelu 27, 783 71 Olomouc, Czech Republic. FAU - Pecinkova, Petra AU - Pecinkova P AD - Department of Cell Biology and Genetics, Faculty of Science, Palacky University, Slechtitelu 27, 783 71 Olomouc, Czech Republic. FAU - Li, Hao AU - Li H AD - Department of Medicine and Genetics, Albert Einstein College of Medicine, Bronx, NY 10461, USA. FAU - Anzenbacher, Pavel AU - Anzenbacher P AD - Department of Pharmacology, Faculty of Medicine and Dentistry, Palacky University, Hnevotinska 5, 779 00 Olomouc, Czech Republic. FAU - Spicakova, Alena AU - Spicakova A AD - Department of Pharmacology, Faculty of Medicine and Dentistry, Palacky University, Hnevotinska 5, 779 00 Olomouc, Czech Republic. FAU - Anzenbacherova, Eva AU - Anzenbacherova E AD - Department of Medical Chemistry and Biochemistry, Faculty of Medicine and Dentistry, Palacky University, Hnevotinska 5, 779 00 Olomouc, Czech Republic. FAU - Chow, Vimanda AU - Chow V AD - Department of Chemistry, York University, 6 Thompson Road, M3J 1L3, ON, Toronto, Canada. FAU - Liu, Jiabao AU - Liu J AD - Department of Molecular Genetics, Donnelly Centre for Cellular and Biomolecular Research, 160 College Street, M5S 3E1, ON, Toronto, Canada. FAU - Krause, Henry AU - Krause H AD - Department of Molecular Genetics, Donnelly Centre for Cellular and Biomolecular Research, 160 College Street, M5S 3E1, ON, Toronto, Canada. FAU - Wilson, Derek AU - Wilson D AD - Department of Chemistry, York University, 6 Thompson Road, M3J 1L3, ON, Toronto, Canada. FAU - Beres, Tibor AU - Beres T AD - Czech Advanced Technology and Research Institute, Palacky University, Slechtitelu 27, 783 71 Olomouc, Czech Republic. FAU - Tarkowski, Petr AU - Tarkowski P AD - Czech Advanced Technology and Research Institute, Palacky University, Slechtitelu 27, 783 71 Olomouc, Czech Republic; Department of Genetic Resources for Vegetables, Medicinal and Special Plants, Centre of the Region Hana for Biotechnological and Agricultural Research, Crop Research Institute, Slechtitelu 27, 783 71 Olomouc, Czech Republic. FAU - Chen, Dajun AU - Chen D AD - Department of Biochemistry, Albert Einstein College of Medicine, Bronx, NY 10461, USA. FAU - Mani, Sridhar AU - Mani S AD - Department of Medicine and Genetics, Albert Einstein College of Medicine, Bronx, NY 10461, USA. Electronic address: sridhar.mani@einsteinmed.edu. LA - eng PT - Journal Article DEP - 20240119 PL - United States TA - Bioorg Chem JT - Bioorganic chemistry JID - 1303703 RN - 0 (Pregnane X Receptor) RN - 0 (Receptors, Cytoplasmic and Nuclear) RN - 0 (Anti-Inflammatory Agents) SB - IM MH - Humans MH - Animals MH - Mice MH - Pregnane X Receptor/agonists MH - Caco-2 Cells MH - *Colitis/chemically induced MH - Receptors, Cytoplasmic and Nuclear MH - Anti-Inflammatory Agents/therapeutic use OTO - NOTNLM OT - Absorption OT - Distribution OT - Metabolism OT - Off targets OT - Pre-clinical evaluation OT - Pregnane X receptor OT - Toxicokinetics COIS- Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. EDAT- 2024/01/22 00:41 MHDA- 2024/02/19 06:43 CRDT- 2024/01/21 18:03 PHST- 2023/11/24 00:00 [received] PHST- 2023/12/25 00:00 [revised] PHST- 2024/01/14 00:00 [accepted] PHST- 2024/02/19 06:43 [medline] PHST- 2024/01/22 00:41 [pubmed] PHST- 2024/01/21 18:03 [entrez] AID - S0045-2068(24)00042-7 [pii] AID - 10.1016/j.bioorg.2024.107137 [doi] PST - ppublish SO - Bioorg Chem. 2024 Mar;144:107137. doi: 10.1016/j.bioorg.2024.107137. Epub 2024 Jan 19.