Expression of Phytocannabinoids in Yeast: a High Yield Platform for Low Abundance Natural Products (Phase II)

AbstractCannabinoids are a class of natural products shown to treat a wide range of diseasesincluding epilepsyone of the most common neurological disordersEpilepsy affects people of all ageswith overmillion epileptics worldwide and overnew cases of epilepsy each year in the United States aloneEven with numerous anti epileptic drugsAEDavailableof patients have uncontrollable seizures that do not respond to treatmentWithout proper seizure control the effects of epilepsy are more pronouncedand in children lack of seizure control not only interrupts early stage brain development but can also lead to social isolationRecentlythe endocannabinoid systemECShas been shown to play a powerful role in a number of neurological diseasesincluding epilepsycreating a new therapeutic approach to control seizures where other AEDs have failedOne of the most effective and promising ECS targeting therapeutics for seizure control is cannabidiolCBDa nonpsychotropic naturally occurring cannabinoidCBD is available over the counter in the United States and pharmaceuticals containing CBD are approved for use in many countriescurrently in Phase III clinical trials in the United StatesCBD is especially attractive since it has been shown to control seizures in patients with pharmaco resistant seizuresespecially childrenwith no significant side effectsAlthough CBD is effective with few side effectspharmaceutical CBD has limited availability and high costprimarily as a result of production shortcomingsThe only source of pharmaceutical or consumer CBD is through agricultural productionpurified from the cannabis or hemp plantFurtherpharmaceutical CBD is slated to cost $$day once approved in the United StatesThese high costs create a treatment gap between poor and wealthy patients and preclude access for the overwhelming majority of epilepticsTo address this issueLibrede has developed a scalable yeast based cannabinoid production technology capable of producing CBD in bulk for andltof the cost of current production methodsallowing a broader population access to life changing medicineIn Phase I workwe successfully engineered yeast to produce CBD using a low cost glucose feedstockIn the Phase II work proposed herewe will optimize CBD production through geneticmetabolicand fermentation engineering to yields andgtmg per liter of cultureLibrede will combine targeted genetic optimization with a screened library approach to efficiently select yeast strains having improved CBD yieldLibrede will also optimize growth and extraction conditions using commercial bioreactorsup toLto improve yield and develop methods compatible with high volume commercial productionAt the conclusion of this work Librede will have a fully developed process for producing and purifying CBD at large scale and low costWidely available low cost CBD has the potential to make a significant difference in the lives of millions of epileptic sufferers worldwide Narrative CannabidiolCBDis a naturally occurring cannabinoid shown to be highly effective in treating pharmacoresistant epilepsy with no side effectsparticularly significant for childrenCurrent plant based production of CBD leads to high therapeutic costs and low availabilityprecluding access to treatment by the majority of the populationTo address this problemLibrede has developed a yeast based biosynthetic CBD production platform capable of large scalelow cost productionIn this work we will optimize and scale CBD production yields