University of Wollongong Research Online University of Wollongong Thesis Collection University of Wollongong Thesis Collections 2012 Design, Synthesis, and Pharmacological Evaluation of Novel Antipsychotic Drugs Based on Olanzapine That Display Reduced Weight Gain and Metabolic Side Effects Somayeh Jafari University of Wollongong Recommended Citation Jafari, Somayeh, Design, Synthesis, and Pharmacological Evaluation of Novel Antipsychotic Drugs Based on Olanzapine That Display Reduced Weight Gain and Metabolic Side Effects, Doctor of Philosophy thesis, School of Health Sciences, University of Wollongong, 2012. http://ro.uow.edu.au/theses/3582 Research Online is the open access institutional repository for the University of Wollongong. For further information contact the UOW Library: [email protected] Design, Synthesis, and Pharmacological Evaluation of Novel Antipsychotic Drugs Based on Olanzapine That Display Reduced Weight Gain and Metabolic Side Effects A thesis submitted in fulfillment of the requirements for the award of the degree of Doctor of Philosophy From The University of Wollongong By Somayeh Jafari Masters of Science (MedChem) Supervisors: Prof. Xu-Feng Huang Dr Francesca Fernandez-Enright School of Health Sciences March, 2012 CERTIFICATION I, Somayeh Jafari, declare that this thesis, submitted in full fulfillment of the requirements for the award of Doctor of Philosophy, in the School of Health Sciences, Faculty of Health and Behavioral Sciences, University of Wollongong, is wholly my own work unless referenced or acknowledged. The document has not been submitted for qualifications at any other academic institution. Somayeh Jafari March, 2012 i TABLE OF CONTENTS CERTIFICATION i TABLE OF CONTENTS ii LIST OF FIGURES vi LIST OF SCHEMES vii LIST OF TABLES viii LIST OF ABBREVIATINS ix PUBLICATIONS xii ACKNOWLEDGEMENTS xiii ABSTRACT xv CHAPTER 1: INTRODUCTION 1 1.1 Contribution of Structure to Therapeutic Actions 2 1.1.1 Phenothiazines and Thioxanthenes 9 1.1.2 Butyrophenones, Diphenylbutylpiperidines and Benzamides 9 1.1.3 Benzisoxazolepiperidines and Indoles 10 1.1.4 Dibenzodiazepines, Thienobenzodiazepines and Dibenzothiazepines 11 1.1.5 Phenylpiperazines and Azapirones 12 1.1.6 Effectiveness of Antipsychotic Drugs in CATIE Study 13 1.2 The Metabolic Side Effects of Antipsychotics 14 1.3 Potential Mechanisms of Antipsychotic-Induced Weight Gain and 19 Discussion of SAR 1.4 Conclusions and Future Prospects 26 1.5 Aims of This Project 27 ii CHAPTER 2: SYNTHESIS AND SAR STUDY OF 29 NOVEL POTENTIAL ANTIPSYCHOTICS 2.1 Background 29 2.2 Synthesis of Novel Olz Derivatives 35 2.3 General Procedures 41 2.3.1 Experimental Procedures, Reagents, and Solvents 41 2.3.2 Microwave Reactions 41 2.3.3 Chromatography 41 2.4 Characterization and Instrumentation 42 2.4.1 1H- and 13C-Nuclear Magnetic Resonance (NMR) Spectroscopy 42 2.4.2 Infrared Spectrometry (IR) 43 2.4.3 Mass Spectrometry (MS) 43 2.4.4 Melting Points 43 2.5 Synthesis of OlzEt 44 2.6 Synthesis of OlzHomo 46 2.7 Synthesis of MeHomo 48 2.8 Characterization of Compound 3 50 2.9 In Vitro Receptor Binding Profiles and SAR Study of Potential 51 Antipsychotics 2.9.1 Method 51 2.9.2 Results 53 2.9.3 Discussion 54 2.9.4 Conclusion 56 iii CHAPTER 3: THE EFFECT OF OLZ,OLZET, 59 AND OlzHOMO ON WEIGHT GAIN AND METABOLIC SIDE EFFECTS 3.1 Background 59 3.2 Materials and Methods 61 3.2.1 Animals and drug treatment regime 3.2.2 Hormonal and post mortem measurement 62 3.2.3 Radioligand binding assay 64 3.2.4 Data analysis 65 3.3 Results 65 3.3.1 Body weight gain, food and water intake 65 3.3.2 Fat deposition 67 3.3.3 Plasma hormonal level 67 3.3.4 Alterations in H1, D2, and 5HT2A receptor densities 68 3.4 Discussion 76 3.4.1 Prevention of weight gain and adiposity following OlzEt and 76 OlzHomo regimes 3.4.2 A study on neurochemical alterations following the treatments 80 with Olz, OlzEt, and OlzHomo 3.5 Conclusion 86 CHAPTER 4: BEHAVIOURAL STUDY: 87 ACOMPARISION BETWEEN OLZ, OLZET, AND OLZHOMO 4.1 Introduction 87 4.2 Materials and Methods 89 4.2.1 Animals and drug treatment regimes 89 4.2.2 Behavioural analysis and post mortem measurement 90 4.2.3 Data analysis 92 4.2.3 Radioligand binding assay 92 4.3 Results 93 4.3.1 Behavioural testing 93 iv 4.3.2 Alteration in D2 and 5HT2A receptor densities 93 4.4 Discussion 97 4.5 Conclusion 100 CHAPTER 5: GENERAL CONCLUSIONS AND FUTURE PROSPECTS 5.1 Conclusion 102 5.2 Future Directions 105 REFERENCES 108 v LIST OF FIGURES Figure 2.1: a) Olz mapped into the proposed H1 agonist 30 pharmacophore model; b) Structure of Olz showing atom numbering Figure 2.2: Chemical structures of compounds 1 (OlzEt), 2, and 3 30 Figure 2.3: Chemical structure of Olz analogues modified at position 34 C-4 Figure 2.4: Chemical structure of dimer 16 36 Figure 2.5: Structural parameters for bioactive conformations of 58 tetracyclic thienobenzodiazepines Figure 3.1: Oral administration of Olz to a female Sprague Dawley 63 rat using the sweet cookie-dough pellet method Figure 3.2: Total body weight gain (g), Total food intake (g), and 70 Energy efficiency in female Sprague Dawley rat treated with Olz, OlzEt, OlzHomo (3 mg/kg or 6 mg/kg), or vehicle (Control) Figure 3.3: Cumulative body weight gain (g), cumulative food 71 intake (g), and cumulative water intake (ml) in female Sprague Dawley rat treated with Olz, OlzEt, OlzHomo, or vehicle (Control) Figure 3.4: Plasma insulin (PM), Leptin (PM), Adiponectin (pg/ml) 72 in female Sprague Dawley rat treated with Olz, OlzEt, OlzHomo, or vehicle (Control) Figure 3.5 (a): Systematic overview of the potential neuropathology 84 underlying increased food intake and obesity pertinent to schizophrenia Figure 3.5 (b): Systematic overview of the potential neuropathology 85 underlying increased food intake and obesity following treatment with Olz Figure 4.1: Rat performing in the open field test 91 vi LIST OF SCHEME Scheme 2.1: Vilsmeier-Haack reaction of Olz 32 Scheme 2.2: Synthesis of compound 2 32 Scheme 2.3: Synthesis of compounds OlzEt 1, OlzHomo 8b, and MeHomo 8a 35 Scheme 2.4: Synthesis of compound 9b 40 vii LIST OF TABLES Table 1.1: Chemical structures and receptor binding affinity profiles of 5 antipsychotics Table 1.2: Preclinical studies on potential future antipsychotics 24 Table 2.1: Various attempted conditions for N-10 protection of Olz 33 Table 2.2: Binding affinities of Olz, compounds 3, OlzEt, OlzHomo, and 54 MeHomo for 5HT2A, D2, and H1 receptors in Sprague Dawley rats Table 3.1: Mean body weight and fat pad mass in female Sprague Dawley 73 rats Table 3.2: H1, D2, and 5HT2A receptor specific densities in the hypothalamus, 74 striatum, and prefrontal cortex of female Sprague Dawley rats Table 3.3: Results of correlation and regression analysis 75 Table 4.1: Open field testing in male Sprague Dawley rats following 2 weeks 95 treatment with Olz, OlzEt , OlzHomo, or vehicle (Control) Table 4.2: D2 and 5HT2A receptor specific densities in the striatum and 96 prefrontal cortex of male Sprague Dawley rats viii LIST OF ABBREVIATIONS 5-HT2 5-hydroxytryptamine A Atypical A Adrenergic receptor AMPK Adenosine monophosphate-activated protein kinase ANOVA Analysis of variance BAT Brown adipose tissue BBB Blood-brain barrier Boc tert-Butoxycarbonyl Calcd Calculated CATIE Clinical antipsychotic trial on intervention effectiveness Cbz Benzyloxycarbonyl CI Chemical ionization Ci Curie CPM Count per minute d Doublet dd Double of doublets D Dopamine receptor DCM Dichloromethane DEPT Distortionless enhancement by polarisation transfer DMF N,N-dimethylformamide DMPK Drug Metabolism/Pharmacokinetics DMSO Dimethylsulfoxide DOPAC Dihyroxyphenylacetic acid dt Double of triplet EDTA Ethyl(carboxymethyl)amino]acetic acid ED50 Median effective dose EI Electron impact EPS Extrapyramidal symptoms equiv Equivalents ESI Electrospray ionization ix GCOSY Gradient correlation spectroscopy GHMBC Gradient heteronuclear multiple bond correlation GHSQC Gradient heteronuclear single quantum correlation Glu Glutamate h Hours H Histamine receptor H Hydrogen/proton HPLC High performance liquid chromatography HRMS High resolution mass spectrometry Hz Hertz IC50 Inhibitory concentration (half maximal) J Coupling constant Kg Kilogram Ki Inhibition constant Kcal Kilo calories LRMS Low resolution mass spectrometry m Multiplet M Molar M Muscarinic receptor MHz Mega hertz min Minutes mL Milliliters mmol Millimole m.p. Melting point MS Mass spectrometry MW Molecular weight m/z Mass to charge ratio nM Nanomolar NMDA N-methyl-D-aspartate acid NMR Nuclear magnetic resonance Olz Olanzapine PCP Phencyclidine x PET Positron-emission tomography pH Potential of hydrogen PPM Part per million PTLC Preparative thin layer chromatogeraphy q Quartet quin Quintet rt Room temperature Ref Reference Rf Retention factor s Singlet SAR Structure activity relationship SDA Serotonin-dopamine antagonist SES 2-(trimethylsilyl) ethyl sulfonyl SREBP-1 Sterol regulatory element binding protein-1 t Triplet T Typical TD Tardive dyskinesia THF Tetrahydofuran TLC Thin layer chromatogeraphy TMS Tetramethylsilane TRIS Tris(hydroxymethyl)aminomethane UCP Uncoupling protein UV Ultraviolet µL Microliter δ Chemical shift (in parts per million, downfield from TMS) xi PUBLICATIONS The following publications and presentations have arisen directly from a work conducted for this thesis. Publication in Referred Journals Jafari S, Fernandez-Enright F, and Huang X-F. Structural contribution of antipsychotic drugs to their therapeutic profiles and methabolic side effects. Journal of Neurochemistry120: 371–384, 2012. (Chapter 1) Jafari S, Bouillon ME, Huang X-F, Pyne SG, and Fernandez-Enright F. Novel olanzapine analogues presenting a reduced H1 receptor affinity and retained 5HT2A/D2 binding affinity ratio. BMC Pharmacology 12: 8, 2012 (Chapter 2) Jafari S, Huang X-F, and Fernandez-Enright F. In vivo pharmacological evaluations of novel olanzapine analogues in rats: potential novel antipsychotic drugs with lower metabolic side effect.