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Doctor of Philosophy (Md) School of Pharmacy Development, safety and efficacy evaluation of actinic damage retarding nano-pharmaceutical treatments in oculocutaneous albinism. J. M Chifamba (R931614G) B. App Chem (Hons), M Phil (Upgraded to D.Phil.), Dip QA, Dip SPC, Dip Pkg Thesis submitted in fulfilment of the requirements for the degree of DOCTOR OF PHILOSOPHY (MD) Main Supervisor: Prof C. C Maponga 1 Associate Supervisor: Dr A Dube (Nano-technologist) 2 Associate Supervisor: Dr D. I Mutangadura (Specialist dermatologist) 3, 4 1School of Pharmacy, CHS, University of Zimbabwe, Zimbabwe 2School of Pharmacy, University of the Western Cape, South Africa 3Fellow of the American academy of dermatology 4Fellow of the International academy of dermatology SCHOOL OF PHARMACY COLLEGE OF HEALTH SCIENCES © Harare, September 2015 i This work is dedicated to the everlasting memory of my dearly departed father, his scholarship, mentorship and principles shall always be my beacon. Esau Jeniel Mapundu Chifamba (15/03/1934-08/06/2015) ii ACKNOWLEDGEMENTS “Art is I, science is we”, Claude Bernard 1813-1878 Any work of this size and scope inevitably draws on the expertise and direction from others. I therefore wish to proffer my utmost gratitude to the following individuals and institutions for their infallible inputs and support. My profound appreciation goes to Prof C C Maponga and Dr A. Dube for crafting, steering and nurturing my interests and research pursuits in nano-pharmaceuticals. Indeed, I have found this discipline to be most intellectually fulfilling. I wish to further acknowledge the mentorship, validation and insight into dermato-pharmacokinetics from the specialist dermatologist Dr D I Mutangadura. Many thanks go to Prof M Gundidza for the contacts, guidance and refereeing in research methodologies, scientific writing and analytical work. I wish to state my Indebtedness to Paidamoyo Kurauvone and the albino welfare organisations in Zimbabwe for embarking on this journey with me and giving me a personal insight into living positively with Albinism. I would also like to acknowledge the related work on skin permeation, spectrophotometry and dermato-pharmacokinetics tape stripping studies done by A O Gamer, Tokumura F et al, Karin Sperling-Vietmeier, and J Lademann et al, which served as efficacy assaying and study guides. I wish to appreciate the assistance and facility from the University of Zimbabwe, School of pharmacy and the HIT, Pharmaceutical technology department. I wish to acknowledge my indebtedness to the sponsorship and financial support by HIT, the University of Zimbabwe research grants and the ICF grant from the Government of Zimbabwe. For instrumental analysis and skin diffusion studies, many thanks go to the Unilever RIC, Croda International ltd Research Laboratories, Mining Research Institute, TRB and Silchem pharmaceuticals for equipment, materials and analytical techniques. Your contributions to this work can never be justly accentuated enough. J M Chifamba iii PUBLICATIONS AND PRESENTATIONS ON RESEARCH-WORK Selected publications from this work in refereed journals 1. Chifamba J, Dube A and Maponga CC: Ex-vivo penetration of nanometric ZnO and TiO2 across actinically damaged porcine skin: Development of an albinistic skin protection treatment. Int J Pharm Sci Res 2015; 6(6): 2339-48.doi: 10.13040/IJPSR.0975-8232.6 (6).2339-48. 2. Chifamba J, Dube A and Maponga CC: Investigation of In-vivo penetration and distribution of nanometric TiO2 in tropical albinistic skin by sequential adhesive tapes stripping. Int J Pharm Sci Res 2015; 6(10): 4181-89.doi: 10.13040/IJPSR.0975-8232.6(10).4181-89 Selected media coverage on this research-work 1. ZTV main news, feature on albinistic treatment development: 15 April 2014: 2. The Herald newspaper, feature article on albinistic treatments: August 2014: 3. Chronicle newspaper: feature article on albinistic treatments: 20 April 2014: 4. Africa on line news, feature article on albinistic treatments: April 2014 5. Indian online news, feature on albinistic treatments: April 2014 6. ZTV main news, feature on Nanomedicine: 07 August 2014: 7. Sunday Mail Newspaper, feature on herbal medicines: 14 September 2014. Selected conference and symposia presentations on research-work 1. Zimbabwe Pharmacists council conference, feature presentation titled: Nanotechnology and herbal based skin treatments: Victoria Falls, March 2015 2. NUST multidisciplinary research conference, paper presentation titled: Emerging technologies impact on dermatological treatments: Victoria Falls November 2014 3. IPSF, feature presentation titled: Emerging technologies in the pharmaceutical industry and novel product development: Victoria Falls June 2014 4. ZPSA annual conference, feature presentation titled: nanotechnology and pharmaceutical product development: ZIPAM march 2014 5. RIE SET, paper presentation and research exhibition titled: Breakthrough in Albinistic treatments: Harare September 2014 6. RIE-SET, paper presentation and research work exhibition titled: Nanotechnology and Indigenous Knowledge systems in skin care week: Harare September 2013 7. Parliamentary portfolio committee on Health presentation titled: Breakthrough in Albinistic treatments: Harare April 2014 iv 8. Parliamentary portfolio committee on Higher and tertiary education, presentation titled: Nanotechnology implementation for socio-economic transformation in Zimbabwe on research-work: Harare August 2014 v ABSTRACT Introduction: There are at least 17 000 Persons living with albinism (PLWA) in Zimbabwe. Oculocutaneous albinism (OCA) is a congenital amelanistic pigmentation disorder that affects all known vertebrates and has no known cure. Melanogenesis is the body’s primary protection from actinic damage, which summarizes all the acute and chronic solar induced adverse dermatological conditions. This impairment therefore makes PLWA highly susceptible to all forms of this damage. Problem statement: Commercial products for actinic damage in PLWA are not readily available. Chemical sunscreens used by PLWA are ineffective and do not treat symptoms of actinic damage. The possible use of promising broad spectrum physical sunscreens in albinistic treatments is hindered by their opaque and un-aesthetic nature. Research hypothesis: A treatment based on nanometric TiO2 and ZnO incorporating the active extracts of A. excelsa, T. emetica and M. flabellifolia will be aesthetic, efficacious and safe in retarding and alleviating all forms of actinic damage in PLWA. Research aims: To develop albinistic actinic damage treatments, using nano TiO2 and ZnO as sun-blocks and incorporating selected herbs. The dermato-pharmacokinetics, stability, efficacy, toxicity and aesthetics of the resultant formulation on albinistic skin types were also investigated in this study. Materials and methods: Emulsion formulation was done according to FDA-CFSAN, COLIPA, and OECD mandated technical guidelines and testing methods. Formulation skin sensitivity were evaluated through Draize ocular and skin sensitivity tests as well as in-vivo patch tests guided by OECD 428/404 technical guidelines and opinion SCCNFP 0750/03. Percutaneous absorption and albinistic skin dermato-pharmacokinetics were evaluated ex-vivo using Franz diffusion tests and sequential adhesive tape stripping respectively according to OECD guidelines 428 and SCCNFP opinions as well as related work done by A O Gamer and Diembeck et al as guides. Analysis for Ti and Zn were done by ICP-AES and Flame AAS respectively. Efficacy and SPF testing was done as per FDA–CFSAN, Colipa and OECD M389/EN mandated test methods. Principal Results: SPF 16, aesthetic and stable emulsions were formulated. Negligible irritation indices for the treatment were recorded for Draize and human patch testing. No percutaneous absorption was observed for ex-vivo diffusion tests and sequential tape stripping tests. Different skin reservoir properties were observed at different skin sites Conclusions: The studies demonstrate, direct evidence that neither Zn nor Ti can penetrate actinic damaged skin regardless of anatomical site and that albinistic dermato-pharmacokinetics are depended on anatomical region and extent of UVR exposure. The high extraction yields and the phyto-constituents of the selected herbs show a correlation with the traditional uses of the plants in traditional medicine. All sensitivity tests showed negligible irritation potential. Based on the foregoing, it is concluded that, incorporation of nanometric TiO2, ZnO and herbs in treatments to retard actinic damage in PLWA is feasible, aesthetic, efficacious, and commercializable and does not pose any health risk. vi CONTENTS INTRODUCTION ............................................................................................................................................. 1 1.1 HUMAN ALBINISM ........................................................................................................................ 2 1.1.1 UVR and sunscreens .............................................................................................................. 4 1.1.2 Natural protection against UVR ............................................................................................ 7 1.1.3 Sunscreens and Sun filters for UVR ....................................................................................... 8 1.1.4 Physical sunscreens ............................................................................................................... 9 1.1.5 Geographical UVR variations impact on cosmeceuticals ...................................................
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