Hydroxypropyl β cyclodextrin hpcd

LCX001 was provided by the Laboratory of Computer-Aided Drug Design & Discovery, Beijing Institute of Pharmacology and Toxicology (Xiao et al., 2016 (link)). Fentanyl (Sigma, Beijing, China) and TH-030418 were administered to generate opioid-induced respiratory depression models. TH-030418 (N-methyl-7a-[(R)-1-hydroxy-1-methyl-3-(thien-3-yl)-propyl]-6,14-endo-ethanotetrahydronororipavine) was a thienorphine derivative, synthesized by our institute. Compared with morphine or dihydroetorphine, TH-030418 displayed non-selective and high binding affinity to all subtypes of opioid receptors (Wen et al., 2011 (link)). TH-030418 also exerted a strong and long-acting analgesic effect. Ampakines, LCX001 and CX614 (Tocris, Bristol, United Kingdom), were dissolved in a 20% hydroxypropyl-β-cyclodextrin (HPCD; Sigma-Aldrich, Beijing, China) 0.45% saline solution. Propofol was purchased from AstraZeneca (Viale Dell’industria, Italy). Pentobarbital (Nembutal; Merck, Beijing, China), Fentanyl, TH-030418, and morphine hydrochloride (Qinghai Pharmaceutical Factory, Xining, China) were dissolved in 0.45% saline solution at room temperature and used at different doses. The volume of each dose was 1 ml/kg for SD rats and 10 ml/kg for KM mice.

Hydrogenated soybean phosphatidylcholine (HSPC), 1, 2-distearoyl-sn-glycero -3-phosphoethanolamine-N-[methoxy(polyethyleneglycol)-2000] (DSPE-PEG2000) were purchased from Lipoid (Germany). Cholesterol and fluorescein labeled PEG-DSPE were from Avanti Polar Lipids (AL, USA). All-trans retinoic acid (ATRA) was obtained from Shandong Liangfu Pharmaceutical Co., Ltd. Other reagents including ethanol, calcium acetate, sodium chloride, potassium chloride and sucrose were of analytical reagent grades and purchased from Sinopharm (Shanghai, China). Fetal bovine serum and hydroxypropyl-β-cyclodextrin (HPCD) were from Sigma-Aldrich (Steinheim, Germany). Acetonitrile and methanol (HPLC grade) were from Thermo Fisher Scientific (Waltham, USA). Therapeutic mouse anti-PD-1 were purchased from BIO X CELL™(USA), InVivoMAb Antibodies,anti-mouse PD-1(CD279,Clone: 29F.1A12).