Creation of new varieties and extracts of hemp for medicinal purposes.

Subsidised by the Innovation and Development Agency of Andalusia (IDEA) through its Incentive Programme for the Innovation and Development of Business.

Code
360135
Started
October 11, 2012
Ended
December 30, 2014

Summary

The general objective of this project was to produce knowledge and to innovate methods to hybridise and cultivate Hemp as a medicinal plant and methods to extract its active substances, procuring the most suitable intellectual protection for all the pursued objectives.

The project was divided into four primary milestones, which in turn were divided into several steps:

Milestone 1: Adaptation of the industrial facility for plant cultivation and commissioning of hybridisation and cultivation laboratories and extraction laboratories. Creation of a method to analyse the active compounds in plant varieties.

Milestone 2: Creation of new dioecious “monocannabinoid” varieties of hemp for medicinal purposes and their registration with the Community Plant Variety Office (CPVO) and/or patenting.

Milestone 3: Creation and patenting of methods of analysis and extraction to produce extracts of medicinal interest.

Milestone 4: Creation and patenting of extracts and their compositions, obtained from the new varieties. Chemical evaluation of the vegetable raw material and extracts obtained from the hybrids proposed for registration, and evaluation of the biological activity of the extracts proposed for registration.

Results

Modalities to cultivate CBD (cannabidiol) and CBG (cannbigerol) chemotypes in high tunnels were studied and found to be a good approach to produce vegetable raw material of varieties with CBD and CBG chemotypes at a commercial level, as well as all the chemotypes characterised by a THC content of <0.2%.

Chromatogram of CBG chemotype obtained by GC-FID:

creation-of-new-varieties-1.jpg#asset:41

The most significant result obtained by this project was the registration with the CPVO of the SARA hemp variety, which has a CBD (cannabidiol) chemotype, under entry number 2015/0098.

Extraction specialisation, such as: classic extraction with organic solvents using maceration with stirring, extraction with organic solvents assisted by auxiliary energies, extraction with supercritical CO2 (in collaboration with the NOVALINDUS platform) and cold extraction with liquefied gas using butane.

Development of an analytical method to determine active substances of the most interest, such as cannabinoids, flavonoids and terpenes, by means of HPLC-APCI.

Figure showing determination of cannabinoids, flavonoids and terpenes by means of HPLC-ACPI.

creation-of-new-varieties-2.jpg#asset:42

1. Linalool oxide (I), 2. Linalool oxide (II), 3. Canniprene, 4. Citral, 5. Pulegone, 6. Dembinobin, 7. Borneol + Eucaliptol, 8. Terpineol, 9. Linalool, 10. Fenchol, 11. CBDV, 12. CB, 13. Caryophyllene , 14. Myrcene, 15. Camphene, 16. Ocimene, 17. Nerolidol, 18. CBD, 19. RHCV, 20. Farnesol, 21. Limonene + carene, 22. Guaiol, 23. CBG, 24. Pinene, 25. CBDA, 26. CBCV, 27. CBN, 28. CBGA, 29. A9THC, 30. CA, 31. A8THC, 32. CBC, 33. THCA.

Study of the biological activity of extracts produced by the varieties to be registered with the CPVO, by means of an in vitro screening test platform; this study covered cytotoxic, neuroprotective, anti-inflammatory, antioxidant, metabolic and angiogenic activity, in addition to ligand binding platforms that interact with CB1 and CB2 cannabinoid receptors and TPVR-1 vanilloid receptors.

Collaborations

- Irrigation Natural Resource Area Research Group at the Las Torres-Tomejil Centre of the Agrarian and Fishing Technologies Research and Education Institute of Andalusia (IFAPA). Collaboration to analyse the productive response of different hemp varieties cultivated with medicinal purposes under different irrigation doses, sowing densities and planting dates in Western Andalusia.

- Research Group BIO304 “Immunopharmacology and Molecular Virology” at the University of Cordoba. Collaboration to conduct in vitro biological activity of plant extracts produced in the project.

- Research group FQM-227 “Analytical platform for metabolomics” at the University of Cordoba. Collaboration to fine-tune methods to extract plant active compounds for their subsequent identification and quantification.