Stopping cocaine before it gets to the brain: new frontiers in treatment CELAC Symposium “Progress and Challenges in Scientific Research on Treatments, Pharmacological Strategies and Vaccines against Drug Addiction” 1 Marilyn E. Carroll 2 2 3 Collaborators: Stephen Brimijoin , Yang Gao , Robin Parks , 1 1 Natalie E. Zlebnik , Justin J. Anker 1Department of Psychiatry, University of Minnesota, Minneapolis, MN 2Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic, Rochester, MN 3Regenerative Medicine Program, Ottawa Health Research Institute, Ottawa, Ontario, Canada NIDA Tested Medications for Treatment of Cocaine Dependence Amantadine Disulfiram Lofexidine Propanolol Lys-dex Divalproex Lobeline Reboxetine amphetamine Dronabinol LY544344 Risperidone Aripriprazole* Fluoxetine* Mecamylamine RTI compounds Atomoxetine Gabapentin Memantine Selegiline Baclofen GBR12909 Methamphetamine Sertraline Buprenorphine GCP44352 Methylphenidate Tiagabine Bup/naloxone Gammavinyl GABA Methadone Topiramate Bupropion Hydromorphone Modafinil Vigagatrin Buspirone LAAM N-acetyl-aspartate Venlafaxine Clonidine L-Dopa/Carbidopa Naltrexone x 2 Yohimbine DHEA L-tryptophan Olanzapine … and More Desipramine Pergolide d-Amphetamine Progesterone Dextrometorphan 3 studies of cocaine hydrolase (CocH) treatment to acutely block: 1. Cocaine self-administration (progressive ratio) 2. Cocaine escalation 3. Cocaine reinstatement 3 studies of CocH viral vector (VEC) treatment to chronically block: 1. Cocaine reinstatement 2. Cocaine-induced locomotor sensitization with cocaine vaccine (VAC) 3. Cocaine self-administration (VEC + VAC) 3 studies of cocaine hydrolase (CocH) treatment to acutely block: 1. Cocaine self-administration (progressive ratio) 2. Cocaine escalation 3. Cocaine reinstatement 3 studies of CocH viral vector (VEC) treatment to chronically block: 1. Cocaine reinstatement 2. Cocaine-induced locomotor sensitization with cocaine vaccine (VAC) 3. Cocaine self-administration (VEC + VAC) BChE - cocaine hydrolase (CocH) - genetically engineered from human BChE A199S/F227A/S287G/A328W/Y332G 10000 F227AS/S287G/A328W/Y332A CocH 1000 cat A199S/S287G/A328W/Y332G 100 cocaine k cocaine A328W/Y332A 10 wild type BChE 1 1 2 3 4 5 (2002) (2003) (2004) (2008) Successive Mutation Generations Molecular interception by enzyme (“Pac-man model”) CocH drastically accelerates cocaine metabolism in rats unprotected subject brain 2500 blood 2000 control rats cocaine 1500 enzyme- 1000 enzyme-treated subject pretreated 500 plasma cocaine brain 0 0 30 60 90 120 blood cocaine Gao et al, Chemico-Biol. Interact. 2010 Brimijoin et al. Neuropsychopharmacol 33: 2715-2725, 2008 CocH rescues rats from cocaine-induced seizures Brimijoin et al. Neuropsychopharmacol 33: 2715-2725, 2008 Locomotor activity apparatus CocH prevents cocaine-induced locomotor activity in mice 70 60 50 40 No BChE Wildtype 30 CocH No Cocaine 20 Beam Crossings Beam 10 0 0 10 20 30 40 50 60 70 Time (min) Brimijoin et al. Neuropsychopharmacol 33: 2715-2725, 2008 Cocaine hydrolase treatment requires enzyme activity to suppress cocaine-stimulation in mice Cholinesterase inhibitor, iso-OMPA, blocked enzyme activity inhibitor restores stimulation inhibitor alone - no effect in presence of enzyme ** ** 500 ** 400 ** ** ** Distance 300 traveled 200 distance distance traveled 100 0 saline CocH CocH + control iso-OMPA iso-OMPA cocaine Carroll et al., 2012 IV drug self-administration apparatus Phases of Drug Abuse Process = 3 studies Acquisition Short vs. Long Access Extinction Reinstatement Drugs Responding Sal Operant Responding Operant Short Access Time (~ 2 - 3 months) Modified from de Vries (1998) and Ahmed and Koob (1998) Cocaine self-administration during short access (2 h) under a progressive-ratio (PR) schedule to assess motivation ** 15 b ** ** CocH enzyme (vs saline) ++ reduced PR performance ++ 12 ++ for 1 session 9 6 3 Mean Mean Infusions (±SEM) 0 0 (Sal) 2 1 4 Albu-CocH Dose (mg/kg) B D A B D A B D A Before, During, After CocH Carroll et al. Psychopharmacology, 2011 Escalation of cocaine self-administration during long access (6 h) d Saline or enzyme treatment 700 For first 7 days 2 mg/kg CocH 600 Saline # 500 ** # ** CocH enzyme (vs saline) increased 400 cocaine infusions for first 4 (of 7) days 300 # # 200 Mean Mean (±SEM) Infusions 100 0 0 3 6 9 12 15 18 21 Days Carroll et al. Psychopharmacology, 2011 Reinstatement (relapse) responses with CocH enzyme no effect on reinst. 4 days after CocH SEM) Responses SEM) CocH acutely ± blocked reinst. Mean ( (E) Enzyme pretreatment (C) cocaine priming injection (S) Saline pretreatment (A) amphetamine priming injection Brimijoin et al. Neuropsychopharmacology 2008 3 studies of cocaine hydrolase (CocH) treatment to acutely block: 1. Cocaine self-administration (progressive ratio) 2. Cocaine escalation 3. Cocaine reinstatement 3 studies of CocH viral vector (VEC) treatment to chronically block: 1. Cocaine reinstatement 2. Cocaine-induced locomotor sensitization with cocaine vaccine (VAC) 3. Cocaine self-administration (VEC + VAC) Adenoviral gene delivery for long-term actions Hydrolase transduction raises cocaine-hydrolase activity in plasma up to 1,000,000-fold 10000 4500 units 1000 100 10 1 .1 CocE activityCocE (mU/ml) .01 0.0045 units .001 1x 3x 5x 10x 5x Control CocE AME359 Year-long generation of CocH with helper-dependent adenoviral vector (hdAD) by gene transfer 10 0 0 hdAD 10 0 10 (mU/ml) 1 first-generation vector 0. 1 injected enzyme cocaine hydrolase activity hydrolase cocaine 0. 0 1 0 10 0 20 0 30 0 40 0 days after injection Ottawa, Mayo Gao and Brimijoin J Pharmacol Exp Ther 330:449-457, 2009 Gene transfer of hydrolase blocks fosB induction in neostriatum in rats given repeated cocaine treatment MW 134 FosB 82 41 32 AME + C S + S E + C S + C CocH 4 3 2 1 0 saline onlysaline AME empty cocaine induction Gao and Brimijoin J Pharmacol Exp Ther 330:449-457, 2009 Cocaine clearance in blood plasma in rats treated 2 weeks earlier with CocH vector or saline controls Plasma cocaine minimal in CocH vector-treated group Plasma cocaine (uM) cocaine Plasma Anker et al., Biol. Psychiatry, 2011 CocH vector reinstatement procedure 1 injection of CocH vector or saline 1 day before extinction P hase ACQ MAINT EXT Reinstatement (8) Protracted Reinstatement (days) (~10) (~10) (14) S C S C S C S A S C S C S C S C S C S C S C S C S A Dose C 0.4 mg/kg, i.v. C (5, 10, 15 ); A (2) C (10) C (10); A (2) (mg/kg, i.p.) Weeks N/A N/A 3 5 6 7 8 12 16 20 24 post vector Blood draws for CocH levels weekly then monthly Anker et al., Biol. Psychiatry, 2011 Maintenance Extinction Males Females Anker et al. Biol. Psychiatry, 2011 Reinstatement after CocH vector (week 3) by cocaine priming dose 15 16 17 18 19 20 21 22 23 24 Days post CocH vector injection Anker et al., Biol. Psychiatry, 2011 Reinstatement responding after CocH vector (weeks 5 - 8) Control > CocH vector Minimal responding on days of saline (S) or no priming injections (1-5) Anker et al., Biol. Psychiatry, 2011 Active lever reinstatement responding # * group differences (0.01, 0.05) Anker et al., Biol. Psychiatry, 2011 Active lever reinstatement responding # * group differences (0.01, 0.05) CocH blood levels (no differences across weeks) SEM) ± Mean ( Mean CocH Vmax (mU/ml)CocH Anker et al., Biol. Psychiatry, 2011 Active lever reinstatement responding Inactive lever responding - control for general # * group differences behavioral suppression no group differences Anker et al., Biol. Psychiatry, 2011 Active lever responding first 3 days after return to operant chamber for monthly reinstatement testing * Day 1 > Days 2 and 3 no group differences Anker et al., Biol. Psychiatry, 2011 Spontaneous locomotor behavior during 3 45 min sessions - - CocH vector vs control (no cocaine administered) no group differences CocH vector did not suppress locomotor activity. Anker et al., Biol. Psychiatry, 2011 3 studies of cocaine hydrolase (CocH) treatment to acutely block: 1. cocaine self-administration (progressive ratio) 2. cocaine escalation 3. cocaine reinstatement 3 studies of CocH viral vector (VEC) treatment to chronically block: 1. cocaine reinstatement 2. cocaine-induced locomotor sensitization with cocaine vaccine (VAC) 3. Cocaine self-administration (VEC + VAC) Enhanced interception with Enzyme & Vaccine brain unprotected subject blood cocaine brain enzyme-treated blood cocaine brain antibody-treated blood cocaine brain enzyme plus antibody blood cocaine Locomotor activity apparatus Each session = Saline - locomotor chamber (45 min) then cocaine (10 mg/kg) - locomotor chamber (45 min) Day 8 Locomotor activity Sensitization Coc > Sal > V+V Day 1 vs 8 Control Yes Yes Yes VAC Yes Yes blocked VEC Yes Yes Yes V + V blocked blocked Carroll et al., 2012 Mean beam breaks over 45 min following cocaine injections across the 8 days of cocaine treatment V + V > VAC, VEC, and Control in reducing locomotor behavior Carroll et al., 2012 Challenge = saline or cocaine injected 15 days after the 8 treatment days Day 24 Locomotor activity Sensitization Coc > Sal < Control Day 1 vs 23 Control Yes Yes VAC Yes Yes blocked VEC Yes Yes Yes V + V Yes blocked Yes Carroll et al., 2012 High dose cocaine in mice and VEC + VAC combinations Effect on locomotor behavior 100 mg/kg cocaine 120 mg/kg, divided (60) 10 min apart CocH + VAC (AB) > VAC, CocH, SAL and Control in reducing locomotor behavior Carroll et al., 2012 VEC + VAC effects on cocaine (0.4 mg/kg) self-administration during 2-hr sessions