Article Figures & Data
Figures
- Fig. 1.
Structure of fentanyl (A) and F (B).
- Fig. 2.
Experiment 1. F-KLH reduces fentanyl-induced antinociceptive effects and increases serum fentanyl concentrations in mice. Vaccination with F-KLH significantly reduced fentanyl-induced hotplate antinociception by 60% (mean ± S.E.M.) (A) and increased serum fentanyl concentrations (mean ± S.D.) (B) compared with controls 30 minutes after a 0.05 mg/kg, s.c., dose of fentanyl. Numbers above bars represent the percent difference from controls. **P < 0.01; ***P < 0.001 compared with controls using unpaired t tests with Welsh’s correction. n = 6/group.
- Fig. 3.
Experiment 2. Selectivity and pharmacokinetic efficacy of F-KLH in rats. (A) Vaccination with F-KLH significantly reduced fentanyl-induced hotplate antinociception by 93% 30 minutes after a 0.035 mg/kg, s.c., dose of fentanyl. (B and C) F-KLH had no effect on heroin- or oxycodone-induced antinociception 30 minutes after a 1 or 2.25 mg/kg dose of heroin or oxycodone, respectively. Serum fentanyl concentrations were significantly increased (D) and brain fentanyl concentrations were significantly decreased (E) compared with controls 4 minutes after a 1-minute 0.05 mg/kg, i.v., infusion of fentanyl. Numbers above bars represent the percentage difference from controls. *P < 0.05; - *P < 0.01; ***P < 0.001 compared with controls. Mean ± S.D. (D and E), mean ± S.E.M. (A–C); n = 12/group (A, D, and E) and n = 6/group (B and C).
- Fig. 4.
Experiments 3 and 4. Fentanyl dose-response and F-sKLH effects on hotplate antinociception, respiratory depression, and bradycardia in rats. Fentanyl was administered subcutaneously every 15 minutes at increasing doses in nonimmunized rats, and the doses listed are the cumulative dose received. (A) Effect of fentanyl on hotplate antinociception. Latency to respond is capped at 60 seconds. Naloxone (0.1 mg/kg, s.c.) was administered 15 minutes after the final fentanyl dose. (B) Effect of fentanyl on respiratory depression measured as SaO2. (C) Effect of fentanyl on heart rate. **P < 0.01; ***P < 0.001 for the difference between values compared with baseline. (D) Vaccine effects on fentanyl-induced antinociception. (E) Vaccine effects on fentanyl-induced respiratory depression measured as SaO2. (F) Vaccine effects on fentanyl-induced decreases in heart rate. *P < 0.05; **P < 0.01; ***P < 0.001 for differences from baseline within groups. #P < 0.05; ##P < 0.01; ###P < 0.001 for the difference between groups at each dose. There were no differences between groups in latency to respond, SaO2, or heart rate after naloxone treatment. Mean ± S.D.; n = 8/group.
- Fig. 5.
Experiment 4; F-sKLH alters fentanyl distribution in serum and to the brain. F-sKLH vaccination increases serum (A) and decreases brain fentanyl (B) distribution by 73% 30 minutes after receiving a cumulative 0.1 mg/kg, s.c., fentanyl dose. Numbers above bars represent the percent difference from controls. Mean ± S.D., ***P < 0.001 compared with controls using unpaired t tests with Welch’s correction.
- Scheme 1.
Synthetic pathway to hapten [8]. (a–g) Reactants: 2-(Boc-amino)ethylbromide, K2CO3, acetonitrile, 80°C, 88% (a); aniline, AcOH, NaBH3CN, CH2Cl2, 80°C, 86% (b); propionyl chloride, DIPEA, CH2Cl2, rt, 95% (c); TFA/CH2Cl2 (2/8-v/v), rt, quantitative (d); glutaric anhydride, pyridine, CH2Cl2, rt, quantitative (e); (Gly)4-OtBu, HBTU, DIPEA, CH2Cl2, rt, 74%; TFA/CH2Cl2 (2/8-v/v), rt, 92% (f); TFA/CH2Cl2 (2/8-v/v), rt, 92% (g).
