Article Figures & Data
Figures
- Fig. 1.
Structure of PF-5190457 and PF-6870961. PF-5190457 is converted to PF-6870961 through the addition of a hydroxy group to the piperidine ring. This reaction occurs via cytosolic enzymes, namely xanthine and aldehyde oxidases.
- Fig. 2.
Inhibition of hGOAT by GSHR1a inverse agonist PF-5190457, metabolite PF-6870961, and hGOAT inhibitor NSM-48. (A) Effect of PF-5190457 at 10–195 μM and PF-6870961 at 10–1000 μM in comparison with known GOAT inhibitor NSM-48 at 10–1000 μM. Percent hGOAT activity = octanoylation of ghrelin mimetic GSSFLCNH2 peptide in presence of inhibitor expressed as a percentage of that in the absence of inhibitor and is normalized to vehicle condition. (B) Structure of NSM-48 provided for reference. Data represent mean and individual values from independent experiments. n = 2–4.
- Fig. 3.
Inhibition of GHSR1a-induced IP mobilization and β-arrestin mobilization by PF-5190457, PF-6870961, and LEAP-2. Concentration response assay to assess inhibitory potency of compounds at GHSR1a expressed on HEK293 cells (A) Inhibition of constitutive GHSR1a inositol phosphate (IP) mobilization in the presence of increasing concentrations of PF-5190457, PF-6870961, or LEAP-2. (B) Inhibition of GHSR1a IP mobilization as a percentage of the maximal ghrelin-induced response in the presence of increasing concentrations of PF-5190457, PF-6870961, or LEAP-2. (C) Inhibition of constitutive GHSR1a β-arrestin recruitment in the presence of increasing concentrations of PF-5190457, PF-6870961, or LEAP-2. (D) Inhibition of GHSR1a β-arrestin recruitment as a percentage of the maximal ghrelin-induced response in the presence of increasing concentrations of PF-5190457, PF-6870961, or LEAP-2. Data represent mean ± S.E.M. n = 3. β-arrestin = β-arrestin 1 and β-arrestin 2.
- Fig. 4.
Effect of PF-5190457, PF-6870961, and JMV2959 on food intake in satiated and fasted male and female rats. PF-5190457 effect on food intake in satiated (A) and fasted (B) male and female rats 1 hour post-injection relative to baseline. PF-6870961 reduced food intake in satiated (C) and fasted (D) male and female rats 1 hour post-injection. JMV2959 reduced food intake in satiated (E) but not fasted (F) male and female rats 1 hour post-treatment. (G) Structure of JMV2959, PF-5190457, and PF-6870961 included for reference. Data represent mean ± S.E.M. *P < 0.05, **P < 0.01, difference from vehicle. n = 10, ○ = male rats, ▲= female rats.
- Fig. 5.
Lack of effect of PF-6870961 on food intake in satiated and fasted male and female GHSR KO rats. PF-6870961 (40 mg/kg) vs. vehicle effect on food intake in satiated (A) and fasted (B) male and female rats 1 hour post-injection. Data represent mean ± S.E.M. p = ns, difference from vehicle. n = 10, ○ = male rats, ▲= female rats.
- Fig. 6.
Time concentration profile of plasma PF-5190457 and PF-6870961 levels in humans after repeated administrations of PF-5190457. Geometric mean plasma concentration-time pharmacokinetic profile of (A) the parent drug PF-5190457 and (B) the metabolite PF-6870961 after repeated dosing of PF-5190457 (0 mg, 50 mg, and 100 mg twice daily) in our placebo-controlled phase 1b human laboratory study in male and female individuals with heavy alcohol drinking. Dosing phase: −3000 to 0 minutes; alcohol session phase: 0 to 1440 minutes. n = 14. Note: The Y-axis of (B) is expanded to show more detail.
- Fig. 7.
Scatterplots of self-reported food craving vs. PF-5190457 or PF-6870961 plasma concentration in humans. Scatterplots depicting PF-5190457 (ng/ml) (top, green) and PF-6870961 (ng/ml) (bottom, purple) vs. GFCQ-S food craving measures during visit 3 (where subjects received 100 mg twice daily PF-5190457). n = 12. CR, cue reactivity.
Tables
Hill Slope Ki (nM) 95% CI Lower Limit (nM) 95% CI Upper Limit (nM) PF-5190457 Human GHSR 1.061 2.96 2.64 3.33 Rat GHSR 0.947 3.94 2.88 5.39 Dog GHSR 1.251 4.88 4.23 5.64 PF-6870961 Human GHSR 0.789 73.6 52.4 103 Rat GHSR 0.811 239 218 262 Dog GHSR 0.929 217 186 253 - TABLE 2
Inhibition of GHSR1a-induced IP and β-arrestin mobilization by LEAP-2, PF-5190457, and PF-6870961
Normalized to Constitutive GHSR1a Activity Normalized to 100% Ghrelin-Induced Activity LEAP-2 PF-5190457 PF-6870961 Ghrelin LEAP-2 PF-5190457 PF-6870961 IP Bottom 7.738 −0.93520 −7.797 53.95 5.869 −0.6585 −4.356 Top 98.81 98.61 100.1 97.09 54.17 61.78 59.26 Hill Slope −1.966 −1.073 −0.7264 1.192 −1.795 −1.039 −0.7219 EC50 (M) — — — 6.58 × 10−10 — — — IC50 (M) 4.77 × 10−9 6.76 × 10−9 3.01 × 10−7 — 5.14 × 10−9 6.87 × 10−9 2.752 × 10−7 Span 91.07 99.54 107.9 43.14 48.30 62.44 63.62 β-Arrestin Bottom −2.582 0.000 0.000 16.12 17.97 17.91 26.20 Top 99.65 100.0 100.0 102.5 −0.5703 1.769 3.128 Hill Slope −1.154 −1.000 −1.000 1.00 −1.00 −1.00 −1.00 EC50 (M) — — — 1.56 × 10−8 — — — IC50 (M) 2.05 × 10−8 3.39 × 10−9 1.10 × 10−9 — 1.98 × 10−8 7.23 × 10−9 1.26 × 10−9 Span 102.2 100.0 100.0 86.40 −18.54 −16.14 −23.07 —, not applicable.
- TABLE 3
Baseline characteristics of participants in analysis of plasma concentrations of PF-5190457 and PF-6870961
Sample Size N = 12 Sex, N (%)
Male
Female
11 (91.7)
1 (8.3)Race, N (%)
African American
White
11 (91.7)
1 (8.3)Smoking Status, N (%)
Smoking
Nonsmoking
9 (75.0)
3 (25.0)BMI in kg/m2 (Mean ± S.E.M.) 26.9 ± 1.3 Education in Years (Mean ± S.E.M.) 12.1 ± 1.0 Age in Years (Mean ± S.E.M.) 40 ± 3.8 90-Day DDDa (Mean ± S.E.M.) 11.3 ± 3.8 BMI, body mass index; DDD, drinks per drinking day.
aFrom 90-day Timeline Followback (TLFB).
- TABLE 4
Correlations between self-reported food craving and PF-5190456 and PF-6870961 plasma concentration
Compound Blood Draw Timepoint GFCQ-S Timepoint Pearson Correlationa P Value Sample Size (N) PF-5190457 Day 1: T2 − T1 vs. Day 1: T2 − T1 −0.116 0.719 12 Day 2: T2 − T1 vs. Day 2: T2 − T1 0.153 0.635 12 Day 2 T2 vs. Day 2: CR2 − CR1 0.431 0.162 12 Day 2 T2 vs. Day 2: CR3 − CR1 0.022 0.945 12 PF-6870961 Day 1: T2 − T1 vs. Day 1: T2 − T1 0.112 0.728 12 Day 2: T2 − T1 vs. Day 2: T2 − T1 −0.095 0.768 12 Day 2 T2 vs. Day 2: CR2 − CR1 −0.015 0.963 12 Day 2 T2 vs. Day 2: CR3 − CR1 0.000 1.000 12 CR, cue reactivity.
aPearson correlation values calculated between individual metabolite concentrations and GFCQ-S scores at different time points during visit 3 (subjects received 100 mg twice daily PF-5190457).
Data Supplement
- Supplemental Data -
Supplementary Appendix S1. Inclusion and Exclusion Criteria
Supplementary Table 1. % Inhibition of Off-Target Binding and Enzymatic Activity of PF-6870961
Supplementary Table 2. %Inhibition of Dopamine, Serotonin, and Opioid Receptors and Related Transporters
Supplementary Table 3. Mean Plasma Concentration of PF-5190457 and PF-6870961
Supplementary Figure S1. PF-51904575 (30 mg/kg) had no effect on spontaneous locomotion in rats as measured by distance traveled (m) (left) and average speed (m/s) (right) in an open field test.
- Supplemental Data -
