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Pheroid® Technology As a Tool to Change the Administration Route of Selected Pharmaceuticals from Intravenous to Oral

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Pheroid® Technology As a Tool to Change the Administration Route of Selected Pharmaceuticals from Intravenous to Oral Pheroid® technology as a tool to change the administration route of selected pharmaceuticals from intravenous to oral J Kleynhans orcid.org/0000-0001-9393-8855 B.Pharm, M.Sc (Pharmaceutical Chemistry) Thesis submitted for the degree Doctor of philosophy in Pharmaceutics at the North-West University Promoter: Prof AF Grobler Co-promoter: Prof JR Zeevaart Co-promoter: Prof MM Sathekge Graduation: October 2018 Student number: 21090955 DEDICATION Dedicated to my dad, Johannes Petrus Kleynhans (1946/12/22 – 2017/07/30), my biggest supporter who was denied by others to witness this final triumph of our journey just before we crossed the finish line. i INSPIRATION Want U is my lamp, o Here! En die Here laat my duisternis opklaar. Want met U loop ek ʼn bende storm, met my God spring ek oor ʼn muur. Psalm 18:29-30 ….it used to be so simple, once upon a time. Because the universe was full of ignorance all around and the scientist panned through it like a prospector crouched over a mountain stream, looking for the gold of knowledge among the gravel of unreason, the sand of uncertainty and the little whiskery eight-legged swimming things of superstition. Occasionally he would straighten up and say things like “hurrah, I’ve discovered Boyle’s Third Law.” And everyone knew where they stood. But the trouble was that ignorance become more interesting, especially big fascinating ignorance about huge and important things like matter and creation, and people stopped patiently building their little houses of rational sticks in the chaos of the universe and started getting interested in the chaos itself – partly because it was a lot more easier to be an expert on chaos, but mostly because it made really good patterns you could put on a t-shirt. Terry Prachett, Witches Abroad ii PERSONAL ACKNOWLEDGEMENTS • First and foremost, I have to thank my Father in heaven for this opportunity and the courage to pursue it till the end. • My mother and sister, words will never describe what you mean to me. • I would like to thank my supervisor Prof. Anne Grobler and my co-supervisors Prof. Jan Rijn Zeevaart and Prof. Mike Sathekge for the opportunity to be part of such an amazing team. Thank you all for understanding the struggle and motivating me at the end when I wanted to give up on studies and life. • Prof. Thomas Ebenhan for assisting me with gaining an understanding of the field of Nuclear Medicine by always being available for answering questions. • Internetainers Rhett & Link for their Youtube channel Good Mythical Morning- thank you for bringing fun into my life during the though stages. • Linkin Park, Three Days Grace, One Republic, Imagine Dragons, Twenty One Pilots, Sia and Audioslave for providing anthems to inspire me during the writing process. • CX fitness and Virgin Active for providing me with the facilities to unload my stress. • All my office friends, in no particular order. Andri, Henri and Ruan (office of 2016). Drieke, Stephanie, Linné, Bianca, Jennifer, Riaan, Palesa and Lerato (office of 2017). Janie, Vusi, Amanda, Johncy and Cathryn (students of Necsa and SBAH). Kobus, Jean and Henrico (Clicks pharmacy). • My other friends for listening to my complaining: Sanet, Nkuli, Madelein and Pieter. • The staff of LCL-Cynologics, thank you for your amazing assistance and kindness, the time spend with you in the beautiful Mauritius, I will treasure always. • Anisa Wadee for providing me with the opportunity to locum part-time to keep the pot cooking. All the rest of the students and staff at Clicks, you have made it a pleasure to work during weekends. iii ACKNOWLEDGEMENTS AND DECLARATION I Janke Kleynhans, hereby declare that this thesis is a record of my own work (except where citations or acknowledgments indicate otherwise) and that the study in part or as a whole has not been submitted to any other university. I would like to acknowledge the following individuals or organizations for their contributions to my study: • Financial assistance was received from the National Research Foundation (NRF) of South Africa as well as the Nuclear Technologies in Medicine and Biosciences Initiative (NTemBI). Opinions expressed and conclusions arrived at, are those of the authors and are not necessarily to be attributed to the NRF and NTeMBI. Financial aid was also provided by the North-West University and the DST/NWU PCDDP. • The North-West University for the facilities provided during all 9 years of studying at this institution. • Dr. Matthew Glyn for providing assistance with all the Confocal Laser Scanning Microscopy analysis and Mr. Lesley Masetle for providing assistance with all the Malvern Mastersizing and zeta-potential measurement. The patient scanning for the clinical trial was performed by Ms. Dalene van Wyk at Steve Biko Academic Hospital. • All the statistical analysis was performed by Prof. Faans Steyn from the Statistical Consultation Services of the North-West University. • Part of the data (oral acute and subchronic evaluation and AMES test) for the article on the toxicity of the Pheroid® delivery system was provided by Dr. Dale Elgar. • The handling of animals: Mr. Cor Bester, Mr. Kobus Venter, Mr. Hylton Buntting, Ms. Antionette Fick, Mr. Henri Dunn, Dr. Jenna Moonsamy, Mr. Deenoo Sathyam, Dr. Alexander Espitalier Noel, Mr. Neville Fitzroy and Dr. Stallone Tendai Terrera. • Ms. Liezl Marie Scholtz for agreeing to be the study monitor for the clinical trials in humans. • The International Atomic Energy Agency (IAEA) for provision of a grant to participate in the International Conference on Integrated Medical Imaging in Cardiovascular Diseases (IMIC 2016) and the for the honour of receiving the European Association of Nuclear Medicine (EANM) award for presenting distinguished work. This has been an opportunity of a lifetime. https://conferences.iaea.org/indico/event/100/contribution/103. iv ABSTRACT Key terms: Pheroid®, radiotracers, insulin, preclinical evaluation, clinical trial, toxicity testing The Pheroid® drug delivery system can change the route of administration from the parenteral route to the oral route. This system is therefore investigated as a safe alternative formulation that can also contribute to better patient compliance. Pheroid® technology is currently on the verge of being applied in the clinical environment and an in-depth evaluation of this system’s toxicity is provided as an original research article (submitted to Toxicology Reports). This is a prerequisite for any registration as a new pharmaceutical entity. The oral formulation of 99mTechnetium methyl diphosphonate demonstrated potential in previous evaluations in Sprague Dawley rats as an alternative to intravenous injections of this radiopharmaceutical. This radiopharmaceutical was selected for evaluation based on clinical need (providing bone scans to patients contra-indicated for injections) as well as the high availability of 99mTc as radiotracer. A clinical trial was designed as a hybrid Phase I/II clinical trial with 16 volunteers. The trial was performed according to Good Clinical Practise regulations, and three patients were enrolled before the study was terminated due to lack of efficacy. Valuable information regarding the Pheroid® delivery system was gained and this application of Pheroid® will be refined and pursued further in the future. A review article is provided (submitted to the Journal of Controlled Release) regarding the application of drug delivery systems in nuclear medicine. This article provides insight into the shortcomings of nuclear medicine that can be addressed by the utilization of drug carrier systems such as Pheroid®. A preclinical evaluation of the pharmacokinetics (for a 5 hour period) as well as a basic toxicology analysis, of oral insulin (entrapped in Pheroid®) was performed on Cynomolgus monkeys. Insulin was formulated in pro-Pheroid® capsules as well as a Pheroid® emulsion and administered through the oral route. The efficacy was evaluated based on a drop in blood glucose levels. A blood clinical biochemistry analysis was also performed to gain information regarding the physiological impact of this system (with the entrapped insulin). The pro-Pheroid® formulations lacked efficacy, but the Pheroid® emulsion was effective and demonstrated a longer-acting effect when compared to the short acting subcutaneous control insulin. The positive results gained in this study indicate that further investigations should be launched. This thesis successfully demonstrated the safety of the Pheroid® delivery system for future applications, provided a summary regarding the nuclear application thereof and showed potential in second non-rodent model for the insulin Pheroid® formulation. v OPSOMMING Sleutelterme: Pheroid®, radioaktiewe merkers, insulien, toksisiteits bepaling, prekliniese evaluering, kliniese proef, Die Pheroid® geneesmiddelafleweringssisteem kan die roete van toediening van die parenterale roete tot orale roete verander. Hierdie sisteem word ondersoek as ʼn veilige alternatiewe formulering wat ook kan bydrae tot beter pasiënte aanvaarding van terapie. Pheroid® tegnologie is op die punt van implementering in die kliniese omgewing en ʼn in diepte evaluering van die sisteem se toksisiteit word verskaf as oorspronklike navorsingings artikel (ingestuur vir publikasie aan Toxicology Reports). Die toets van toksisiteit is ʼn voorvereiste vir enige registrasie as ʼn nuwe farmaseutiese entiteit. Die orale formulering van 99mTechnetium metieldifosfonaat in Pheroid® het voorheen in pre- kliniese toetse in Sprague Dawley rotte om ʼn effektiewe
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