Identification of Glycochenodeoxycholate 3‐O‐Glucuronide and Glycodeoxycholate 3‐O‐Glucuronide as Highly Sensitive and Specific OATP1B1 Biomarkers

Abstract

The aim of this study was to investigate the sensitivity and specificity of endogenous glycochenodeoxycholate and glycodeoxycholate 3-O-glucuronides (GCDCA-3G and GDCA-3G) as substrates for organic anion transporting polypeptide 1B1 (OATP1B1) in humans. We measured fasting levels of plasma GCDCA-3G and GDCA-3G using liquid chromatography-tandem mass spectrometry in 356 healthy volunteers. The mean plasma levels of both compounds were ~ 50% lower in women than in men (P = 2.25 × 10 −18 and P = 4.73 × 10 −9). In a microarray-based genome-wide association study, the SLCO1B1 rs4149056 (c.521T>C, p.Val174Ala) variation showed the strongest association with the plasma GCDCA-3G (P = 3.09 × 10 −30) and GDCA-3G (P = 1.60 × 10 −17) concentrations. The mean plasma concentration of GCDCA-3G was 9.2-fold (P = 8.77 × 10 −31) and that of GDCA-3G was 6.4fold (P = 2.45x10 −13) higher in individuals with the SLCO1B1 c.521C/C genotype than in those with the c.521T/T genotype. No other variants showed independent genome-wide significant associations with GCDCA-3G or GDCA-3G. GCDCA-3G was highly efficacious in detecting the SLCO1B1 c.521C/C genotype with an area under the receiver operating characteristic curve of 0.996 (P < 0.0001). The sensitivity (98-99%) and specificity (100%) peaked at a cutoff value of 180 ng/mL for men and 90 ng/mL for women. In a haplotype-based analysis, SLCO1B1*5 and *15 were associated with reduced, and SLCO1B1*1B, *14, and *35 with increased OATP1B1 function. In vitro, both GCDCA-3G and GDCA-3G showed at least 6 times higher uptake by OATP1B1 than OATP1B3 or OATP2B1. These data indicate that the hepatic uptake of GCDCA-3G and GDCA-3G is predominantly mediated by OATP1B1. GCDCA-3G, in particular, is a highly sensitive and specific OATP1B1 biomarker in humans.

The Role of Uptake and Efflux Transporters in the Disposition of Glucuronide and Sulfate Conjugates

“…Organic cation uptake transporters, OCTs and OCTNs, do not appear to interact with conjugate metabolites. For example, neither glucuronides of estradiol, glycochenodeoxycholate (GCDCA) or morinidazole, nor sulfates of estrone, ethinyl-estradiol or morinidazole are substrates for OCTs ( Han et al, 2010a , 2010b ; Zhong et al, 2014 ; Bi et al, 2019 ; Neuvonen et al, 2021 ). Furthermore, several glucuronides, such fevipiprant-G, baicalein-7-G and epacadostat-G, were reported not to inhibit OCTs and MATEs, although uremic toxin indoxyl-G inhibits OCT2 ( Xu et al, 2013 ; Cheung et al, 2017 ; Zhang et al, 2017 ; Poller et al, 2019 ).…”

“…Most importantly, bilirubin glucuronides are high affinity substrates for both OATP1B1 and OATB1B3 and thus, these transporters are partly responsible for controlling plasma levels of conjugated bilirubin ( König et al, 2000 ; Cui et al, 2001 ; van de Steeg et al, 2010 , 2012 ). Other endogenous conjugate substrates for these transporters include bile acid and steroid conjugates, such as ursodeoxycholate-AG, GCDCA-G and GCDCA-S, glycodeoxycholate-G (GDCA-G) and estradiol-17-G ( Takehara et al, 2017 ; Bi et al, 2019 ; Zhou et al, 2019 ; Neuvonen et al, 2021 ). Glucuronides of several drugs, such as ezetimibe, gemfibrozil and sorafenib, are transported by OATP1B1 and OATP1B3, which may contribute to the enterohepatic recycling of these drugs by directing the excretion of metabolites to the bile and feces, instead of excretion into the urine ( Oswald et al, 2008 ; Hirouchi et al, 2009 ; Zimmerman et al, 2013 ; Kimoto et al, 2015 ; Bins et al, 2017 ) .…”

“…In contrast, estrone-S was identified as a rather good substrate for OAT2 ( Kobayashi et al, 2005 ; Xu et al, 2013 ; Mathialagan et al, 2018 ; Bi et al, 2019 ). Several glucuronides and sulfates of natural and endogenous compounds, such as quercetin-S and resveratrol-G, estradiol-17-G and glucuronides of GCDCA and GDCA are not transported by OAT2 ( Wong et al, 2011a , 2012 ; Xu et al, 2013 ; Bi et al, 2019 ; Neuvonen et al, 2021 ).…”

Environmental Chemical Contribution to the Modulation of Bile Acid Homeostasis and Farnesoid X Receptor Signaling

“…BAs and some of their conjugates glycochendodeoxycholate-3-sulfate (GCDCA-S) and chenodeoxycholate-24-glucoronide (CDCA-24G) have been proposed as endogenous biomarkers of hepatic transporter Solute carrier organic anion transporter family member 1B1 (OATP1B1), Solute carrier organic anion transporter family member 1B3 (OATP1B3), BSEP and NTCP in drug development (Watanabe et al, 2015;Takehara et al, 2017;Chu et al, 2018;Neuvonen et al, 2021). The use of conjugated BAs as endogenous biomarkers to inform potential drug-drug interactions (DDI) is a huge stride towards better informing risk assessment in drug development and could also potentially inform DDI management strategies.…”

“…Even this effect may be beneficial in treating neonates with hyperbilirubinemia, the non-therapeutic effects of FXR activation in other patient population may need to be evaluated. Furthermore, BAs and their conjugates have also been explored for their role as endogenous biomarkers as substrates for transporter DDI (Watanabe et al, 2015;Takehara et al, 2017;Chu et al, 2018;Neuvonen et al, 2021).…”