Semisynthetic bile acid FXR and TGR5 agonists: physicochemical properties, pharmacokinetics, and metabolism in the rat.
We report on the relationship between the structure-pharmacokinetics, metabolism, and therapeutic activity of semisynthetic bile acid analogs, including 6α-ethyl-3α,7α-dihydroxy-5β-cholan-24-oic acid (a selective farnesoid X receptor [FXR] receptor agonist), 6α-ethyl-23(S)-methyl-3α,7α,12α-trihydroxy-5β-cholan-24-oic acid (a specific Takeda G protein-coupled receptor 5 [TGR5] receptor agonist), and 6α-ethyl-3α,7α-dihydroxy-24-nor-5β-cholan-23-sulfate (a dual FXR/TGR5 agonist). We measured the main physicochemical properties of these molecules, including ionization constants, water solubility, lipophilicity, detergency, and protein binding. Biliary secretion and metabolism and plasma and hepatic concentrations were evaluated by high-pressure liquid chromatography-electrospray-mass spectrometry/mass spectrometry in bile fistula rat and compared with natural analogs chenodeoxycholic, cholic acid, and taurochenodexycholic acid and intestinal bacteria metabolism was evaluated in terms of 7α-dehydroxylase substrate-specificity in anaerobic human stool culture. The semisynthetic derivatives detergency, measured in terms of their critical micellar concentration, was quite similar to the natural analogs. They were slightly more lipophilic than the corresponding natural analogs, evaluated by their 1-octanol water partition coefficient (log P), because of the ethyl group in 6 position, which makes these molecules very stable toward bacterial 7-dehydroxylation. The hepatic metabolism and biliary secretion were different: 6α-ethyl-3α,7α-dihydroxy-5β-cholan-24-oic acid, as chenodeoxycholic acid, was efficiently conjugated with taurine in the liver and, only in this form, promptly and efficiently secreted in bile. 6α-Ethyl-23(S)-methyl-3α,7α,12α-trihydroxy-5β-cholan-24-oic acid was poorly conjugated with taurine because of the steric hindrance of the methyl at C23(S) position metabolized to the C23(R) isomer and partly conjugated with taurine. Conversely, 6α-ethyl-3α,7α-dihydroxy-24-nor-5β-cholan-23-sulfate was secreted in bile unmodified and as 3-glucuronide. Therefore, minor structural modifications profoundly influence the metabolism and biodistribution in the target organs where these analogs exert therapeutic effects by interacting with FXR and/or TGR5 receptors.
Dipartimento di Chimica "G. Ciamician" Alma Mater Studiorum-University of Bologna, Italy (A.R., C:C:, S.S.); Dipartimento di Chimica e Tecnologia del Farmaco, Università degli Studi di Perugia, Italy (R.P., A.G.); Dipartimento di Scienze Mediche e Chirurgiche, Alma Mater Studiorum-University of Bologna, Italy (F.N., M.M.); TES Pharma S.r.l., Corciano, Italy (R.P., D.P., F.D.F., C.C.); Intercept Pharmaceuticals, Inc., New York, New York; and Dipartimento di Farmacia e Biotecnologie, Alma Mater Studiorum-University of Bologna, Bologna, Italy (R.A.) aldo.roda@unibo.it.
Aldo
A
Roda
Dipartimento di Chimica "G. Ciamician" Alma Mater Studiorum-University of Bologna, Italy (A.R., C:C:, S.S.); Dipartimento di Chimica e Tecnologia del Farmaco, Università degli Studi di Perugia, Italy (R.P., A.G.); Dipartimento di Scienze Mediche e Chirurgiche, Alma Mater Studiorum-University of Bologna, Italy (F.N., M.M.); TES Pharma S.r.l., Corciano, Italy (R.P., D.P., F.D.F., C.C.); Intercept Pharmaceuticals, Inc., New York, New York; and Dipartimento di Farmacia e Biotecnologie, Alma Mater Studiorum-University of Bologna, Bologna, Italy (R.A.).
Roberto
R
Pellicciari
Dipartimento di Chimica "G. Ciamician" Alma Mater Studiorum-University of Bologna, Italy (A.R., C:C:, S.S.); Dipartimento di Chimica e Tecnologia del Farmaco, Università degli Studi di Perugia, Italy (R.P., A.G.); Dipartimento di Scienze Mediche e Chirurgiche, Alma Mater Studiorum-University of Bologna, Italy (F.N., M.M.); TES Pharma S.r.l., Corciano, Italy (R.P., D.P., F.D.F., C.C.); Intercept Pharmaceuticals, Inc., New York, New York; and Dipartimento di Farmacia e Biotecnologie, Alma Mater Studiorum-University of Bologna, Bologna, Italy (R.A.).
Antimo
A
Gioiello
Dipartimento di Chimica "G. Ciamician" Alma Mater Studiorum-University of Bologna, Italy (A.R., C:C:, S.S.); Dipartimento di Chimica e Tecnologia del Farmaco, Università degli Studi di Perugia, Italy (R.P., A.G.); Dipartimento di Scienze Mediche e Chirurgiche, Alma Mater Studiorum-University of Bologna, Italy (F.N., M.M.); TES Pharma S.r.l., Corciano, Italy (R.P., D.P., F.D.F., C.C.); Intercept Pharmaceuticals, Inc., New York, New York; and Dipartimento di Farmacia e Biotecnologie, Alma Mater Studiorum-University of Bologna, Bologna, Italy (R.A.).
Flavia
F
Neri
Dipartimento di Chimica "G. Ciamician" Alma Mater Studiorum-University of Bologna, Italy (A.R., C:C:, S.S.); Dipartimento di Chimica e Tecnologia del Farmaco, Università degli Studi di Perugia, Italy (R.P., A.G.); Dipartimento di Scienze Mediche e Chirurgiche, Alma Mater Studiorum-University of Bologna, Italy (F.N., M.M.); TES Pharma S.r.l., Corciano, Italy (R.P., D.P., F.D.F., C.C.); Intercept Pharmaceuticals, Inc., New York, New York; and Dipartimento di Farmacia e Biotecnologie, Alma Mater Studiorum-University of Bologna, Bologna, Italy (R.A.).
Cecilia
C
Camborata
Dipartimento di Chimica "G. Ciamician" Alma Mater Studiorum-University of Bologna, Italy (A.R., C:C:, S.S.); Dipartimento di Chimica e Tecnologia del Farmaco, Università degli Studi di Perugia, Italy (R.P., A.G.); Dipartimento di Scienze Mediche e Chirurgiche, Alma Mater Studiorum-University of Bologna, Italy (F.N., M.M.); TES Pharma S.r.l., Corciano, Italy (R.P., D.P., F.D.F., C.C.); Intercept Pharmaceuticals, Inc., New York, New York; and Dipartimento di Farmacia e Biotecnologie, Alma Mater Studiorum-University of Bologna, Bologna, Italy (R.A.).
Daniela
D
Passeri
Dipartimento di Chimica "G. Ciamician" Alma Mater Studiorum-University of Bologna, Italy (A.R., C:C:, S.S.); Dipartimento di Chimica e Tecnologia del Farmaco, Università degli Studi di Perugia, Italy (R.P., A.G.); Dipartimento di Scienze Mediche e Chirurgiche, Alma Mater Studiorum-University of Bologna, Italy (F.N., M.M.); TES Pharma S.r.l., Corciano, Italy (R.P., D.P., F.D.F., C.C.); Intercept Pharmaceuticals, Inc., New York, New York; and Dipartimento di Farmacia e Biotecnologie, Alma Mater Studiorum-University of Bologna, Bologna, Italy (R.A.).
Francesca
F
De Franco
Dipartimento di Chimica "G. Ciamician" Alma Mater Studiorum-University of Bologna, Italy (A.R., C:C:, S.S.); Dipartimento di Chimica e Tecnologia del Farmaco, Università degli Studi di Perugia, Italy (R.P., A.G.); Dipartimento di Scienze Mediche e Chirurgiche, Alma Mater Studiorum-University of Bologna, Italy (F.N., M.M.); TES Pharma S.r.l., Corciano, Italy (R.P., D.P., F.D.F., C.C.); Intercept Pharmaceuticals, Inc., New York, New York; and Dipartimento di Farmacia e Biotecnologie, Alma Mater Studiorum-University of Bologna, Bologna, Italy (R.A.).
Silvia
S
Spinozzi
Dipartimento di Chimica "G. Ciamician" Alma Mater Studiorum-University of Bologna, Italy (A.R., C:C:, S.S.); Dipartimento di Chimica e Tecnologia del Farmaco, Università degli Studi di Perugia, Italy (R.P., A.G.); Dipartimento di Scienze Mediche e Chirurgiche, Alma Mater Studiorum-University of Bologna, Italy (F.N., M.M.); TES Pharma S.r.l., Corciano, Italy (R.P., D.P., F.D.F., C.C.); Intercept Pharmaceuticals, Inc., New York, New York; and Dipartimento di Farmacia e Biotecnologie, Alma Mater Studiorum-University of Bologna, Bologna, Italy (R.A.).
Carolina
C
Colliva
Dipartimento di Chimica "G. Ciamician" Alma Mater Studiorum-University of Bologna, Italy (A.R., C:C:, S.S.); Dipartimento di Chimica e Tecnologia del Farmaco, Università degli Studi di Perugia, Italy (R.P., A.G.); Dipartimento di Scienze Mediche e Chirurgiche, Alma Mater Studiorum-University of Bologna, Italy (F.N., M.M.); TES Pharma S.r.l., Corciano, Italy (R.P., D.P., F.D.F., C.C.); Intercept Pharmaceuticals, Inc., New York, New York; and Dipartimento di Farmacia e Biotecnologie, Alma Mater Studiorum-University of Bologna, Bologna, Italy (R.A.).
Luciano
L
Adorini
Dipartimento di Chimica "G. Ciamician" Alma Mater Studiorum-University of Bologna, Italy (A.R., C:C:, S.S.); Dipartimento di Chimica e Tecnologia del Farmaco, Università degli Studi di Perugia, Italy (R.P., A.G.); Dipartimento di Scienze Mediche e Chirurgiche, Alma Mater Studiorum-University of Bologna, Italy (F.N., M.M.); TES Pharma S.r.l., Corciano, Italy (R.P., D.P., F.D.F., C.C.); Intercept Pharmaceuticals, Inc., New York, New York; and Dipartimento di Farmacia e Biotecnologie, Alma Mater Studiorum-University of Bologna, Bologna, Italy (R.A.).
Marco
M
Montagnani
Dipartimento di Chimica "G. Ciamician" Alma Mater Studiorum-University of Bologna, Italy (A.R., C:C:, S.S.); Dipartimento di Chimica e Tecnologia del Farmaco, Università degli Studi di Perugia, Italy (R.P., A.G.); Dipartimento di Scienze Mediche e Chirurgiche, Alma Mater Studiorum-University of Bologna, Italy (F.N., M.M.); TES Pharma S.r.l., Corciano, Italy (R.P., D.P., F.D.F., C.C.); Intercept Pharmaceuticals, Inc., New York, New York; and Dipartimento di Farmacia e Biotecnologie, Alma Mater Studiorum-University of Bologna, Bologna, Italy (R.A.).
Rita
R
Aldini
D000818:Animals; D001419:Bacteria / Q000378:metabolism; D001646:Bile / Q000737:chemistry; D001647:Bile Acids and Salts / Q000032:analysis / Q000097:blood / Q000493:pharmacokinetics*; D055598:Chemical Phenomena; D006801:Humans; D008099:Liver / Q000378:metabolism; D051381:Rats; D018160:Receptors, Cytoplasmic and Nuclear / Q000819:agonists*; D043562:Receptors, G-Protein-Coupled / Q000819:agonists*; D000099150:Receptor, Farnesoid X-Activated
D001647:Bile Acids and Salts; D018160:Receptors, Cytoplasmic and Nuclear; D043562:Receptors, G-Protein-Coupled; D000099150:Receptor, Farnesoid X-Activated; C473772:GPBAR1 protein, human