Article Figures & Data
Figures
- Fig. 1.
Chemical structures of dotinurad and commercially available uricosuric agents. Dotinurad (A), benzbromarone (B), lesinurad (C), and probenecid (D).
- Fig. 2.
Concentration-response curve of dotinurad and commercially available uricosuric agents on urate transport by URAT1-overexpressing MDCKII cells. Each value is the mean ± S.D. of urate transport relative to the control (n = 3) and described by GraphPad Prism 7.03 software (GraphPad Software).
- Fig. 3.
Effects of dotinurad and benzbromarone on plasma urate level, FEUA, and urinary urate excretion in Cebus monkeys. Data are represented as change in plasma urate level (A); FEUA at 0–4, 4–8, and 8–24 hours after dosing (B); and the amount of urinary urate excretion for 8 hours after dosing (C). Each value is the mean ± S.D. of four to five animals. *P < 0.05; **P < 0.01 for significant differences from the control at each time point using the Dunnett’s multiple comparison test.
- Fig. 4.
Effect of dotinurad and other hypouricemic agents on the plasma concentration of sulfasalazine in Sprague-Dawley rats. Hypouricemic agents or their vehicle as control (0.5% MC) was orally administered to Sprague-Dawley rats fasted for 18 hours. After 30 minutes, sulfasalazine was orally administered. Blood samples were collected at 0.25, 0.5, 1, 2, 4, 8, and 12 hours after sulfasalazine administration. Plasma samples were deproteinated, and sulfasalazine concentrations were measured using liquid chromatography–tandem mass spectrometry (LC-MS/MS). Each value is the mean ± S.D. of three to four animals. **P < 0.01, significant differences from the 0.5% MC control group after Dunnett’s multiple comparison test.
- Fig. 5.
Effect of dotinurad and other uricosuric agents on the plasma concentration of adefovir in Sprague-Dawley rats. Hypouricemic agents or their vehicle as control (0.5% MC) was orally administered to Sprague-Dawley rats fasted for 18 hours. After 30 minutes, adefovir was intravenously administered. Blood samples were collected at 0.083, 0.25, 0.5, 1, 2, and 4 hours after adefovir administration. Plasma samples were derivatized, and adefovir concentrations were measured using HPLC. Each value is the mean ± S.D. of six animals. *P < 0.05; **P < 0.01, significant differences from the 0.5% MC control group after Dunnett’s multiple comparison test.
- Fig. 6.
Comparison of the potency of selective and nonselective urate reabsorption inhibitors in decreasing urate pool and the concomitant effects of nonselective inhibitors. (Upper) The usual state of urate handling showing urate excretion from the renal tubules and the small intestine. (A) SURI increases urate excretion from the renal tubules due to URAT1 inhibition, leading to a potent reduction of urate pool. (B) A nonselective inhibitor decreases urate excretion from the renal tubules and/or the small intestine through inhibition of ABCG2 and/or OAT1/3. For a nonselective inhibitor to obtain the same effect at a level similar to SURI, more potent inhibition of URAT1 and additional renal excretion are required. Arrow width represents the quantity of urate handling, which is affected by inhibition of URAT1, ABCG2, and OAT1/3.
Tables
- TABLE 1
IC50 values of dotinurad and commercially available uricosuric agents against urate transport by URAT1 and urate secretion transporters
For measurement of transport activities, MDCKII-URAT1 (passage numbers 3–11), HEK293-OAT1 (passage number 8), HEK293-OAT3 (passage number 5), and HEK293-ABCG2 membrane vesicles (SOLVO Biotechnology) were used. IC50 was calculated using a four-parameter logistic equation. Each value is the mean + S.E.M. (n = 2 to 3).
Test Article IC50 URAT1 ABCG2 OAT1 OAT3 μM μM μM μM Dotinurad 0.0372 + 0.0065 4.16 + 2.49 4.08 + 1.02 1.32 + 0.07 Benzbromarone 0.190 + 0.029 0.289 + 0.074 3.14 + 1.59 0.967 + 0.071 Lesinurad 30.0 + 3.9 26.4 + 4.7 6.99 + 1.94 1.07 + 0.27 Probenecid 165 + 14 433 + 218 10.9 + 0.5 2.37 + 0.94 IC50, half maximal inhibitory concentration.
- TABLE 2
IC50 ratios for ABCG2 and OAT1/3 to URAT1 of dotinurad and commercially available uricosuric agents
Each value was calculated as the IC50 of each compound for ABCG2, OAT1, and OAT3/IC50 for URAT1.
Test Article IC50 Ratio to URAT1 URAT1 ABCG2 OAT1 OAT3 Dotinurad 1 112 110 35.5 Benzbromarone 1 1.52 16.5 5.09 Lesinurad 1 0.880 0.233 0.0357 Probenecid 1 2.62 0.0661 0.0144 - TABLE 3
Pharmacokinetic parameters of dotinurad and benzbromarone in Cebus monkeys
Plasma samples were deproteinated and drug concentrations were measured using HPLC. AUC0–24h was calculated using the trapezoidal rule. Each value is the mean ± S.D. of five animals.
Administered Article Dose Measured Article Pharmacokinetic Parameters Cmax Fu × Cmax Tmax AUC0–24h mg/kg μg/ml μg/ml hours μg⋅hours/ml Dotinurad 1 Dotinurad 2.09 ± 0.78 0.0144 3.2 ± 1.1 20.3 ± 8.6 5 11.4 ± 4.8 0.0800 2.8 ± 1.1 109 ± 53 30 107 ± 22 0.750 2.0 ± 0.0 780 ± 265 Benzbromarone 30 Benzbromarone 20.9 ± 9.0 0.564 2.0 ± 0.0 95.2 ± 28.1 6-Hydroxybenzbromarone 12.7 ± 4.4 — 2.0 ± 0.0 80.9 ± 33.9 —, not calculated; AUC0–24h, area under the plasma concentration-time curve at 0–24 hours; Cmax, maximum plasma concentration; Fu, unbound fraction in plasma; Fu × Cmax, unbound maximum plasma concentration; Tmax, time to reach maximum plasma concentration.
- TABLE 4
Pharmacokinetic parameters of sulfasalazine and effects of concomitant treatment with hypouricemic agents in Sprague-Dawley rats
Pharmacokinetic parameters were calculated using Phoenix WinNonlin. F values were calculated using the following equation: F = (AUC0–inf value of oral administration/20)/(AUC0–inf value of intravenous administration/5) × 100. Values are reported as the mean ± S.D. of three to four animals.
Test Article Dose Cmax T1/2 AUCo–t AUCinf CLT F mg/kg ng/ml hours ng⋅h/ml ng⋅hours/ml ml/hours/kg % 0.5% MC — 422 ± 182 1.49 ± 0.28 665 ± 94 713 ± 90 28,395 ± 3800 2.0 Febuxostat 20 3138** ± 272 1.30 ± 0.13 5004** ± 851 5012** ± 848 4067** ± 607 14.1 Benzbromarone 50 1520** ± 202 1.24 ± 0.46 2858** ± 750 2888** ± 729 7294** ± 1989 8.1 Dotinurad 1.3 415 ± 68 1.88 ± 1.56 708 ± 166 885 ± 264 24,126 ± 7777 2.5 AUCinf, area under the plasma concentration-time curve from zero to infinity; AUC0–t, AUC at 0–12 hours; CLT, total clearance; F, bioavailability; MC, methylcellulose; T1/2, elimination half-life.
↵** P < 0.01, significant differences from the 0.5% MC control group, as analyzed by Dunnett’s multiple comparison test.
Data Supplement
- Supplemental Tables -
Tables S1-S3.
- Supplemental Tables -
