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Neurochemical and Neuropharmacological Studies on a Range Of Neurochemical and Neuropharmacological Studies on a Range of Novel Psychoactive Substances by Barbara Loi Dissertation Submitted to the University of Hertfordshire in partial fulfilment of the requirements of the degree of Doctor of Philosophy Psychopharmacology, Drug Misuse and Novel Psychoactive Substances Research Unit Hertfordshire, UK School of Life and Medical Sciences University of Hertfordshire Date: November 2017 1 Table of contents Table of contents ..................................................................................................................................... 2 List of figures .......................................................................................................................................... 6 List of tables .......................................................................................................................................... 10 Aknowledgements ................................................................................................................................. 11 List of abbreviations ............................................................................................................................. 12 Abstract ................................................................................................................................................. 14 Aims, objectives and hypotheses of the overall PhD project ................................................................ 17 Chapter 1: Novel Psychoactive Substances (NPS) ............................................................................... 20 1.1) NPS - general overview ................................................................................................................. 20 1.2) Epidemiology of NPS drugs .......................................................................................................... 21 1.3) NPS classification .......................................................................................................................... 24 Chapter 2: Index compounds in the present PhD project ...................................................................... 30 2.1) Synthetic cannabinoids .................................................................................................................. 30 2.1.1) Classification of synthetic cannabinoids............................................................................. 30 2.1.2) Third generation synthetic cannabinoids ............................................................................ 31 2.1.3) ‘Spice’ drugs ....................................................................................................................... 35 2.1.4) Epidemiology of synthetic cannabinoids ............................................................................ 36 2.1.5) Social issues represented by synthetic cannabinoids and their effects ............................... 37 2.1.6) Pharmacology of synthetic cannabinoids, focus on their effects on the reward circuit ...... 38 2.1.7) Synthetic cannabinoids and pharmacodynamics ................................................................ 43 2.2 Pipradrol-derivatives ....................................................................................................................... 44 2.2.1) Pharmacological studies on 2-DPMP and D2PM ............................................................... 45 2.2.2) Social issues represented by 2-DPMP and D2PM and their effects ................................... 46 2.3) 5-(2-Aminopropyl)indole (5-IT) .................................................................................................... 47 2.3.1) Social issues represented by 5-IT and its effects ................................................................ 47 2.3.2) Pharmacological studies on 5-IT ........................................................................................ 48 2.4) Aminorex derivatives: focus on 4,4’-DMAR ................................................................................ 49 2.4.1) Pharmacological studies on 4,4’-DMAR ............................................................................ 50 2.4.2) Social issues represented by 4,4’-DMAR and its effects .................................................... 51 2.5) Dieting aid compound 2,4-DNP .................................................................................................... 51 2.5.1) Social issues represented by 2,4-DNP and its effects ......................................................... 53 2.5.2) Pharmacological studies on 2,4-DNP: focus on preclinical studies ................................... 54 Chapter 3: In vitro quantitative autoradiography studies ...................................................................... 54 3.1) In vitro quantitative autoradiography overview............................................................................. 54 2 3.2) Competition binding assay ............................................................................................................ 55 3.3) Autoradiography procedure ........................................................................................................... 56 3.4) Synthetic cannabinoids and autoradiography studies .................................................................... 58 3.4.1) SCs and CB1 receptor autoradiography studies: materials and methods ............................ 60 3.4.2) SCs and CB1 receptor autoradiography studies: results ...................................................... 62 3.4.3) SCs and NMDA receptor autoradiography studies: materials and methods ....................... 69 3.4.4) SCs and NMDAR autoradiography studies: results ........................................................... 70 3.4.5) CB1R and NMDAR autoradiography studies with SCs: discussion ................................... 75 3.5 Synthetic stimulants and DAT autoradiography studies ................................................................. 77 3.5.1) Synthetic stimulants and DAT autoradiography studies: materials and methods ............... 78 3.5.2) Synthetic stimulants and DAT autoradiography studies: results ........................................ 80 3.5.3) Synthetic stimulants and DAT autoradiography studies: discussion .................................. 84 Chapter 4: In vitro fast scan cyclic voltammetry studies ...................................................................... 85 4.1) In vitro fast scan cyclic voltammetry overview ............................................................................. 85 4.2) Fast scan cyclic voltammetry principles ........................................................................................ 87 4.3) BB-22 and FSCV studies: aims ..................................................................................................... 89 4.4) BB-22 FSCV studies: materials and method ................................................................................. 90 4.5) BB-22 FSCV studies: results ......................................................................................................... 93 4.6) BB-22 and FSCV studies: discussion ............................................................................................ 94 Chapter 5: Microdialysis studies ........................................................................................................... 96 5.1) Microdialysis overview ................................................................................................................. 96 5.2) Synthetic cannabinoids and microdialysis studies ....................................................................... 101 5.2.1) Synthetic cannabinoid microdialysis studies: materials and methods .............................. 103 5.2.2) Results for experiment 1: Effect of BB-22 administration on DA transmission in the NAc shell, core and in the mPFCx ...................................................................................................... 106 5.2.3) Results for experiment 2: Role mediated by CB1 receptors on the increase of DA release in the NAc shell DA induced by BB-22 ..................................................................................... 109 5.2.4) Results for experiment 3: Effect of 5F-PB-22, 5F-AKB-48, and STS-135 administration on DA transmission in the NAc shell .......................................................................................... 109 5.2.5) Synthetic cannabinoids and microdialysis studies: discussion ......................................... 112 5.3) 2,4-Dinitrophenol (2,4-DNP) and microdialysis studies ............................................................. 113 5.3.1) 2,4-DNP microdialysis studies: materials and methods ................................................... 114 5.3.2) 2,4-DNP microdialysis studies: results ............................................................................. 116 5.3.3) 2,4-DNP microdialysis studies: discussion ....................................................................... 118 5.4) 2-DPMP and D2PM microdialysis studies .................................................................................. 119 5.4.1) 2-DPMP and D2PM microdialysis studies: materials and methods ................................. 120 5.4.2) 2-DPMP microdialysis experiment: results ...................................................................... 122 3 5.4.3) D2PM microdialysis experiment: results .........................................................................
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