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Clomipramine in Dogs: Pharmacokinetics, Neurochemical CLOMIPRAMINE IN DOGS: PHARMACOKINETICS, NEUROCHEMICAL EFFECTS, AND EFFICACY IN COMPULSIVE DISORDER A Thesis Presented to The Faculty of Graduate Studies of The University of Guelph by CAROLINE J. HEWSON ln partial fulfillment of requirements for the degree of Doctor of Philosophy June, 1997 Q Caroline J. Hewson, 1997 Acquisitions and Acquisitions et Bibliographie SeMces services bibliographiques 395 Wellington Street 395, rue Wellington ûüawa ON K1A ON4 OttawaON K1AON4 Canada Canada The author has granted a non- L'auteur a accordé une licence non exclusive licence aiiowing the exclusive pe~mettantà la National Lîbrary of Canada to Bibliothèque nationale du Canada de reproduce, loan, distn'bute or sell reproduire, prêter, distri'buer ou copies of this thesis in microform, vendre des copies de cette thèse sous paper or electronic formats. la forme de microfichelfilm, de reproduction sur papier ou sur format électronique. The author retains ownership of the L'auteur conserve la propriété du copwght in this thesis. Neither the droit d'auteur qui protège cette thèse. thesis nor substantid extracts from it Ni la thèse ni des extraits substantiels may be printed or otherwise de ceilen ne doivent être imprimés reproduced without the author's ou autrement reproduits sans son permission. autorisation. ABSTRACT Clomipramine in Dogs: Pharmacokinebics, Neurochemical Effects, and EffÎcacy in Compulsive Disorder Caroline Joan Hewson Co-advisors: University of Guelph. 1997 RO Bal& UA Luescher Canine compulsive disorder is a syndrome of abnomal conflict behaviours, possibly associated with central neurochemical dysfunction. There is no proven treatment for the disorder, but the human anti-compulsive drug, clomipramine, has been reported to be effective. Three expenments investigated biological and behavioural effects of clomipramine in dogs. Experïrnent 1 was a pharmacokinetic study. Six dogs received one oral dose (3 mgkg) of clomipramine. After dosing, blood was taken at O, 15, 30,45min. 1, 2, 4. 8, 12h, and then 12-hourly up to 120h. The dogs then received 28 daily doses (3mglkg q 24 h); on d28, they were sampled as before. There was interdog variability in the elimination half-life of clomipramine (1-9h) and its metabolite, desmethylclomipramine (1-4h). Experiment 2 used six other, behaviourally normal dogs to assess the effect of clomipramine (3 mglkg q 24 h PO) on turnover of serotonin, dopamine and norepinephrine, as indicated by concentrations of the respective metabolites in cerebrospinal fiuid (CSF). The experiment had a randomised, placebo-controlled, crossover design. CSF was taken after 1,2,4 and 6 weeks on each treatment: there was a 2-week washout period between treatments. No effect on neurotransrnitter turnover was detected. Experiment 3 was a randomised, placebo-controlled, double-blind, crossover clinical trial of clomipramine (3 rng/kg q 12h PO). Cases of canine compulsive disorder were diagnosed by a clinical ethologist. following telep hone interviews with owners. An independent rater also interviewed owners, using formal criteria to diagnose compulsive disorder. Diagnostic agreement between the ethologist and the criteria was low (~=0.02;n=60). Fifty-one cases were enrolled in the clinical trial. They received each treatment for 4 weeks, with a 2- week washout perÏod in-between. At the end of each treatment, owners rated the severity of the compulsive behaviour using hnro scales of acceptable construct validity (p4.01). Both scales indicated a treatment effect (odds ratio=4; pe0.015, n=50). These experimentç contribute new information about the pharmacokinetics, neurochemical effect and clinical use of clomipramine in dogs. They describe the development of a valid measure of behavioural change, and demonstrate that clomipramine is a useful treatrnent for canine compulsive disorder. ACKNOWLEDGEMENTS My sincere thanks to all those who have made this research possible. In particular, I thank my advisory cornmittee (DE. Ron Ball, Andrew Luescher, Peter Conlon and Joane Parent) for their guidance and good humour. My thanks also ta the Departrnents of Population Medicine, and Animal and Poultry Science for their support. I am deeply grateful to the sponsors of my research. Persona1 funding was provided by the Canadian Commonwealth Scholarship Program and by scholanhips from the University of Guelph and the Ontario Veterinary College. The research funds were provided by Novartis Animal Health, Pet Trust and the Animal Health Trust of Canada. I acknowledge the contribution of al1 the participating dogs and their owners and handlers. Thanks also to the Central Animal Facility. and to Jodie Karrow, Heather Cole, Jacky Adarnek. Craig Moseley and Cindy Leonard, al1 of whom provided excellent technical assistance. To VictorÏa Edge, Mohamed Shoukri and Andrew Milan for statistical advice. To Muriel Burke and al1 in the Clinical Research Building for their support. To al1 who have supported and encouraged me, including fellow students, family and fnends. Special thanks to Christine Power, Tarcia Gennring, Ute Straube, Laura Taylor, Wiil Wistowsky, Janet Schell, John Craven, Dee Britney, Lawrence Cahill. Nicola Pritchard, Alison McDermott, Adrian Glasser, Patncia Harton-McCord, David and Caroline Jordan, Anne Valliant, Don McKeown and Jack Halip. CHAPTER 3 : THE PHARMACOKINETICS OFCLOMIPRAMINEAND DESMFTHYLCLOMIPRAMINE 1N DOGS: PARAMETER ESTIMATES FOLLOWING A SINGLE ORAL DOSE AND 28 CONSECUTIVE DAILY ORAL DOSES OF CLOMIPRAMINE 3.1 lntroduction ...................................................................................................32 3.2 Materials and Methods ................................................................................. 34 3.2.1 Single-dose study .................................................................................. 35 3.2.2 Multipledose study ................................................................................ 35 3.2.3 Handling of samples .............................................................................. 36 3.2.4 Detenination of clomipramine and de~rnethylclorniprarnine~~~~.~-~~~~~~--.36 3 .2.5 Pharmacakinetic analysis ........................ ....**.............*..........*.....*~.... 38 3-3 Results .......................................................................................................... 39 3.4 Discussion .................................................................................................... 52 CHAPTER 4: THE EFFECT OF CLOMIPRAMINE ON THE TURNOVER OF CENTRAL MONOAMINE METABOLITES IN BEHAVlOURALLY NORMAL DOGS Introduction.................................................................................................. -59 4.2 Materials and Methods ................................................................................. 61 4.2.1 Sampling technique .................... .. .................................................... 62 4.2.2 Sample handling .................................................................................. ..63 4.2.2.1 CSF ......... .. ................................................................................... 63 4.2.2.2 Blood ............................................................................................... 64 4.2.3 Chrornatographic Analysis - monoamine metabolites ......................... 65 4.2.3.2 5-HIAA and HVA ............................................................................-66 4.2.3.2 MHPG.............................................................................................. 66 4.2.4 Determination of clomipramine and desmethylclomipramine ............... -67 4-25 Statistical Analysis ................................................................................ -67 4.3 Results .......................................................................................................... 69 4.4 Discussion ......................... .. ...........6 CHAPTER 5: DIAGNOSIS OF CANINE COMPULSIVE DISORDER: EVALUATION OF THE DIAGNOSTIC ACCURACY OF FORMAL CRITERIA Introduction ................................................................................................. ..83 5.2 Materials and Methods ................................................................................. 84 5.2.1 Recruitment of Cases ........................................................................... -84 iii 5.2.2 Diagnostic methods ........................................... ................................. 84 5.2.2.1 Interview #1 .................................................................... -85 5.2.2.2 Interview #2 .................................................................................. 86 5 2.3 Measurement of diagnostic accuracy .................................................... 86 5.3 Results ......................................................................................................... 89 5.3.1 Description of the sample ...................................................................... 89 5.3.1 -1 Sarnple size ..................................................................................... 89 5 .3.1.2 Gender ............................................................................................ 89 5.3.1.3 Age .................................................................................................. 89 5 -3-1 -4 Breed ............................................................................................... 89 5.3.1 -5 Behaviour ..,. ................................................................................... -90 5.3.2
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