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7 protocols using bupropion hydrochloride

1

Synthesis and Characterization of Psychoactive Compounds

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Bupropion hydrochloride and 6-hydroxybupropion were obtained from Toronto Research Chemicals (Toronto, Canada), ibogaine from Ibogaworld, cocaine from Medisca (Montreal, Canada), β-PEA, pinoline, indole, tryptamine, ephedrine, frovatriptan, alprenolol, isoproterenol, evodiamine, and tyramine from Sigma (Oakville, Canada), fluoxetine and amphetamine were from Tocris Bioscience (Bristol, United Kingdom). PAL analogs, RTI analogs, and bicifadine were synthesized at RTI in the Blough laboratory as described previously (Carroll et al., 2009 (link), 2011 (link); Blough et al., 2011 , 2014 (link)). Noribogaine (Batch No. 606950002) was from Deborah Mash, Ph.D. (Obach et al., 1998 ). Ibogamine was obtained from Specs (Zoetermeer, The Netherlands). Sodium phenylbutyrate was purchased from Enzo Life Sciences Inc. (Farmingdale, NY). Ibogaminalog, ibogainealog, noribogainalog, fluorogainalog, and tabernanthalog were synthesized in the lab of David E. Olson (Cameron et al., 2020 (link)).
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2

Methamphetamine, Bupropion, and NAC Interactions

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D-methamphetamine hydrochloride and NAC were purchased from Sigma Chemical (St. Louis, MO) and were dissolved in sterile 0.9% physiological saline. NAC was pH adjusted with NaOH to 7.2 ± 0.2. Bupropion hydrochloride was purchased from Toronto Research Chemicals (Toronto, Canada) and dissolved in sterile water. Methamphetamine was administered intravenously (IV) at a volume of 35.74 μl per infusion. NAC and bupropion were administered intraperitoneally (IP; 1ml/kg injection volume). Methamphetamine, bupropion, and NAC doses were adapted from previous studies (Reichel et al., 2008 , 2011 (link), 2009 (link)).
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3

Nicotine and Bupropion Behavioral Assessment

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Bupropion hydrochloride (Toronto Research Chemicals; Toronto, ON, Canada) and (–)-nicotine hydrogen tartrate (Sigma; St. Louis, MO, USA) were dissolved in 0.9% saline. The pH of nicotine was adjusted to 7.0 ± 0.2 with a dilute NaOH solution. All drugs were administered at 1 ml/kg. Bupropion was administered intraperitoneal (IP); nicotine was injected subcutaneous (SC). Injection routes, drug doses, and time between injection and onset of behavioral assessment were based on previous reports from our laboratory (Besheer et al., 1999 (link); Bevins et al., 2001 (link); Murray and Bevins, 2007 (link); Wilkinson et al., 2010 (link)). Nicotine dose is reported as base, whereas bupropion doses are reported as salt.
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4

Radioligand Binding Assay Protocol

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[3H]-Granisetron (BRL-43694); 85.3 Ci/mmol; 1μCi/μL) was obtained from PerkinElmer Life Sciences, Inc. (Boston, MA). Acetylcholine (ACh), glycine (Gly) and γ–aminobutyric acid (GABA) were purchased from Sigma-Aldrich (St. Louis, MO). Bupropion hydrochloride and hydroxybupropion were purchased from Toronto Research Chemicals, Inc. (North York, Canada); serotonin (5-HT; serotonin creatinine sulfate monohydrate) from Acros Organics (New Jersey, NJ), MDL-7222 from Sigma-Aldrich (St. Louis, MO); protease inhibitor cocktail set III from EMD (Calbiochem; Darmstadt, Germany) and trypsin (TPCK-treated) from Worthington (Lakewood, NJ). Dulbecco’s modified Eagle’s medium/Ham’s F-12 50/50 mix (DMEM/Ham’s F-12) was obtained from Mediatech, Inc. (Herndon, VA) and Geneticin (G-418.SULFATE) was purchased from A. G. Scientific, Inc. (San Diego, CA).
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5

Nicotine, Bupropion, and Varenicline Pharmacology

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Nicotine bitartrate, varenicline tartrate, and Bupropion hydrochloride were dissolved in 0.9% sterile saline. Intravenous nicotine dose (0.03 mg/kg/inf; MP Biomedicals; Solon, OH, USA) and nicotine subcutaneous injection dose (0.4 mg/kg) were chosen based on previous research (Charntikov et al., 2014 (link); Liu et al., 2008 (link)). Bupropion hydrochloride and varenicline tartrate were obtained from Toronto Research Chemicals (Toronto, ON, Canada). Doses and administration protocols were adopted from previous research (Bruijnzeel and Markou, 2003 (link); George et al., 2011 (link); Liu et al., 2008 (link); Shoaib et al., 2003 (link); Wouda et al., 2011 (link)). Nicotine doses are reported as base, whereas bupropion and varenicline doses are reported as salt.
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6

Nicotinic Receptor Ligands: Characterization

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(−)-Nicotine hydrogen tartrate and (−)-cytisine [CYT] were purchased from Sigma (St. Louis, MO, USA). Bupropion hydrochloride [BUP] was purchased from Toronto Research Chemicals (Toronto, ON, Canada). S-(−)-nornicotine fumarate [NOR] was provided by Girindus America, Inc. (Cincinnati, OH, USA). PHA-543613 [PHA], sazetidine-A [SAZ-A], and PNU-120596 [PNU] were gifts from the National Institute on Drug Abuse (Research Triangle Institute, Research Triangle Park, NC, USA). Nicotine doses are reported as free base equivalent, whereas all other compounds are reported in salt form. The vehicle, route of administration, injection volume, and injection-to-placement interval (IPI) for each compound are detailed in Table 1.
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7

Pharmacological Manipulation of Nicotine Responses

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Nicotine bitartrate (MP Biomedicals, OH, USA), varenicline tartrate (Sigma-Aldrich, MO, USA), bupropion hydrochloride (Toronto Research Chemicals, ON, Canada), and yohimbine hydrochloride (Sigma-Aldrich, MO, USA) were dissolved in 0.9 % sterile saline. Intravenous nicotine dose (0.03 mg/kg/inf;) and nicotine subcutaneous injection dose (0.4 mg/kg) were chosen based on previous research [14 (link),17 (link),23 (link)]. All doses and administration protocols were adopted from previous research [11 (link),14 (link),20 (link),33 (link),34 (link)]. Nicotine doses are reported as base, whereas bupropion, varenicline, and yohimbine doses are reported as salt.
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