Bioenzymatic synthesis of THC-v, CBV and CBN and their use as therapeutic agents
The present invention provides methods for producing cannabinoids. More specifically, the invention is directed to the bio-enzymatic synthesis of THC-v, CBV and CBN by contacting a compound according to Formula I with a cannabinoid synthase enzyme. Also described is a system for producing these pharmaceutically important cannabinoids and the use of such cannabinoids as therapeutic agents.
1. A method for producing a compound of Formula III:
wherein R 1 is n-propyl or n-pentyl,
comprising the steps of contacting a compound of Formula I
in a mixture comprising an aqueous buffer and at least one organic solvent selected from the group consisting of methanol, ethanol, propanol, isopropanol, acetone, dimethylsulfoxide, hexane, and ethylacetate
with a cannabinoid synthase to produce a compound of Formula II:
and either
(A1) oxidizing the compound of Formula II to produce a compound of Formula A:
and
(A2) de-carboxylating the compound and of Formula A to produce the compound of Formula III;
OR
(B1) de-carboxylating the compound of Formula II to produce a compound of Formula B:
and
(B2) oxidizing the compound of Formula B to produce the compound of Formula III.
2. The method of claim 1 , wherein the cannabinoid synthase is a tetrahydrocannabinolic acid (THCA) synthase.
3. The method of claim 2 , wherein R 1 is n-pentyl.
4. The method of claim 2 , wherein R 1 is n-propyl.
5. The method of 1 , wherein the compound of Formula II is subjected to steps (A1) and (A2).
6. The method of claim 1 , wherein the compound of Formula II is subjected to steps (B1) and (B2).
7. The method of claim 1 , wherein the de-carboxylating step comprises exposing the compound of Formula A or Formula II to heat, UV light, or a weak base.
8. The method of claim 1 , wherein the oxidizing step comprises exposing the compound of Formula II or Formula B to air, oxygen, gas, an oxidizing agent, an oxidation catalyst, UV-light, or a combination thereof.
9. The method of claim 1 , wherein the organic solvent is dimethylsulfoxide.
10. The method of claim 1 , wherein the aqueous buffer is selected from the group consisting of citrate buffer, phosphate buffer, HEPES buffer, Tris buffer, MOPS buffer, or glycine buffer.
11. The method of claim 1 , wherein R 1 is n-pentyl, the cannabinoid synthase is a tetrahydrocannabinolic acid (THCA) synthase; and
the compound of Formula II is subjected to steps (B1) and (B2).
12. The method of claim 1 , wherein R 1 is n-pentyl, the cannabinoid synthase is a tetrahydrocannabinolic acid (THCA) synthase; and
the compound of Formula II is subjected to steps (A1) and (A2).
13. The method of claim 1 , wherein R 1 is n-propyl, the cannabinoid synthase is a tetrahydrocannabinolic acid (THCA) synthase; and
the compound of Formula II is subjected to steps (B1) and (B2).
14. The method of claim 1 , wherein R 1 is n-propyl, the cannabinoid synthase is a tetrahydrocannabinolic acid (THCA) synthase; and
the compound of Formula II is subjected to steps (A1) and (A2).
15. The method of claim 1 , wherein the concentration of the organic solvent is between 10% and 90% (v/v).
16. The method of claim 1 , wherein the concentration of the organic solvent is between 10% and 30% (v/v).