Development Process for Production of Natural THC and CBD Abstract Δ9-Tetrahydrocannabinol (Δ9-THC) and cannabidiol (CBD) are two major constituents of the cannabis plant that have pharmacological properties with potential therapeutic value in several disease conditions. While Δ9- THC has already been in use clinically for two indications (nausea and vomiting in cancer patients receiving chemotherapy and appetite stimulation for AIDS patients) in an oral soft gelatin capsule formulation, CBD is awaiting to be developed. There is a great need for supply of these two cannabinoids for product development and there is a market for GMP quality material. This proposal is directed toward the development of a process for large scale isolation of these two cannabinoids from crude cannabis extracts that is both economic and capable of producing GMP quality products. This will be accomplished through our discovery that, unlike the free cannabinoids, the protected amino acid esters of the cannabinoids are much easier to purify (separate) on normal phase silica column chromatography and the derivatives are stable enough to be stored until needed. Mild base hydrolysis of the esters generates the free cannabinoids in pure form when needed. The specific aims of this proposal are: 1. Procure cannabis extracts with high THC content (  50% THC) for THC production and high CBD (  50% CBD) for CBD production and to optimize the process of vacuum distillation of the cannabis extracts(s) to enhance the cannabinoid concentration in the distillate. 2. Prepare different t-boc protected amino acid esters of the distillate for chromatographic separation, evaluate the separation of the different cannabinoid derivatives from the different esters using TLC chromatography (normal phase) and select the best system and the best derivative for further evaluation on column chromatography. 3. Evaluate the efficiency of separation of the different components of the derivatized extract (distillate) on small scale (less than gram) quantities and evaluate the recovery of THC from the purified ester through base hydrolysis and analysis of the recovered THC. 4. Optimize the process (which derivative and which chromatographic system) for each target cannabinoid, e.g., THC, CBD. During this phase I the emphasis will be on THC but data will be collected for CBD for further studies during phase II. 5. Scale up the process for isolation of gram quantities of the target cannabinoids. During Phase I application, emphasis will be on the separation of THC and CBD. Other cannabinoids, e.g., THCV and CBG, will be targeted in Phase II. It is anticipated that, at the conclusion of this phase I effort, a process will be in place ready to be validated in phase II to produce GMP quality Δ9-THC and CBD.

Project Narrative This phase I application is designed to develop an efficient and scalable process for the isolation of Δ9-tetrahydrocannabinol (Δ9-THC or THC) and cannabidiol (CBD) from cannabis extract(s). During this effort the process will be brought to the point where GMP validation and production of GMP materials could be accomplished during phase II, making GMP supply of THC and CBD available from ElSohly Laboratories. The ultimate goal is to make GMP supply of THC and CBD available for product development and commercialization.