Cannabis - Erfaringer danner Israel - produktion, forskning. Lumír Ondřej Hanuš Lumir Lab, det Hebraiske Universitet i Jerusalem, Israel Pharma på Tværs 2018 København, Kongeriget Danmark November 3, 2018 Cannabis - experiences form Israel - production, research Lumír Ondřej Hanuš Lumir Lab, Hebrew University, Jerusalem, Israel Pharma på Tværs 2018 Copenhagen, Kingdom of Denmark November 3, 2018 CannabisCannabis sativasativa L.L. (( hemp)hemp) In China 6,000 years ago In China 4700 years ago during the reign of mythical emperor Shen- Nung the first evidence of the use of hemp (chu-ma, female hemp) as medicine to treat: - malaria - gout (arthritis urica) - beri-beri (a nutritional deficit in thiamine) - constipation He deemed chu-ma one of the Superior Elixirs - rheumatism of Immortality - absentmindedness - menstrual problems Scythians an ancient Iranic people of horse-riding nomadic pastoralists 5th century B.C. seeds of Scythian cannabis Pazyryk, Siberia Herodotus of Halicarnassus The History, Book 4 - Melpomene 4.74 Hemp grows in Scythia... 4.75. The Scythians then take the seed of this hemp and creep under the felt coverings, and then they throw the seed upon the stones which have been heated red-hot: and it burns like incense and produces a vapour so think that no vapour-bath in Hellas would surpass it: and the Scythians being delighted with the vapour-bath howl like wolves. John Russell Reynolds British neurologist and physician For the relief of certain kinds of pain, I believe, there is no more useful medicine than Cannabis within our reach. Archives of Medicine 2, 154-160 (1859) Indian hemp, when pure and administered carefully, is one of the most valuable medicines we possess. Lancet 135, 637-638 (1890) University of Illinois UNIVERZITA PALACKÉHO V OLOMOUCI Zdeněk Krejčí (March 25, 1923 – March 31, 1992 ) cannabidiolic acid UNIVERZITA PALACKÉHO V OLOMOUCI František Šantavý (April 23, 1915 – March 27, 1983) 1963; cannabidiol Mechoulam R., Shvo Y.: Tetrahedron 19, 2073–2078 (1963) Šantavý F.: Acta Univ. Palacki. Olomuc., Fac. Med. 35, 5-9 (1964) Yehiel Gaoni Raphael Mechoulam * September 1, 1928 * November 5, 1930 Endocannabinoid system The endocannabinoid system consists of: genes encoding two major cannabinoid receptors, CB1 and CB2, endocannabinoids, and the enzymes involved in the synthesis and degradation of endocannabinoids Cannabinoid receptors Cannabinoid receptors * Two types identified CB1, present in brain and central nervous system, 472 amino acids CB2, present in spleen and immune cells, 360 amino acids * Both types are 7-helix transmembrane spanning receptors. Three extra-cellular and three intra-cellular loops. A glycosylated extra-cellular N-terminal domain, and an intra-cellular C-terminal domain involved in the interaction with the G-protein. 2 1 CB1 CB2 360 Devane, 1988 Munro, 1993 472 Brain regions in which cannabinoid receptors are abundant. __________________________________________________________________ Brain Region Functions Associated with Region __________________________________________________________________ Basal ganglia Movement control Substantia nigra pars reticulata Entopedunculus nucleus Globus pallidus Putamen Cerebellum Body movement, coordination Hippocampus Learning and memory, stress Cerebral cortex, especially cingulate Higher cognitive function frontal, and parietal regions Nucleus accumbens Reward center __________________________________________________________________ Cannabinoid receptors distribution CB1 receptor CB2 receptor cerebral cortex lymphoid organs: hippocampus spleen basal ganglia thymus cerebellum tonsils bone marrow reproductive system leukocytes pancreas macrophages CB1 expression 3.0E+00 2.0E-01 /GAPDH 1 1.0E-01 CB 0.0E+00 cerebellum spleen liver islets of Langerhans GAPDH = glyceraldehyde 3-phosphate CB2 expression dehydrogenase 1.0E-03 1.8E-05 1.2E-05 /GAPDH 2 6.0E-06 CB 0.0E+00 cerebellum spleen liver islets of Langerhans GPR119 (335) GPR18 (331) CB1 (472) GPR55 (319) GPR3 (330) CB2 (360) GPR6 (362) GPR12 (334) PPARa (468) TRPV1 (819) PPARd (441) PPARg (505) Sequence similarities for putative cannabinoid and cannabinoid-like receptors in man. Cannabidiol, a novel inverse agonist for GPR3, GPR6, and GPR12 GPR3, GPR6, and GPR12 are constitutively active, Gs protein- coupled receptor that currently have no confirmed endogenous ligands. GPR3, GPR6, and GPR12 may be involved in physiological processes such as maintenance of oocyte meiotic arrest and brain development, as well as pathological conditions such as metastatic cancer. Cannabidiol (CBD), a major non-psychoactive phytocannabinoid, acted as an inverse agonist to inhibit cAMP accumulation stimulated by the constitutively active GPR3, GPR6, and GPR12. Thus, GPR3, GPR6, and GPR12 are novel molecular targets for CBD. Gs, but not Gi is involved in the inverse agonism of CBD on GPR3, GPR6, and GPR12 . Laun et al., 2018 Tetrahydrocannabinolic acid is a potent peroxisome proliferator-activated receptor gamma (PPARg) agonist with neuroprotective activity Cannabinoid acids are more potent PPARγ agonists and transcriptional activators than their decarboxylated analogues. Crystal Structure of Human PAPRα complex with agonist D9-THCA shows potent neuroprotective activity, which is worth considering for the treatment of Huntington’s disease and possibly other neurodegenerative and neuroinflammatory diseases. These data would strongly suggest Δ9-THCA as a lead structure for the development of novel drugs. Nadal et al., 2017 Endocannabinoids Anandamide William A. Devane,* Lumir Hanuš, Aviva Breuer, Roger G. Pertwee, Lesley A. Stevenson, Graeme Griffin, Dan Gibson, Asher Mandelbaum, Alexander Etinger, and Raphael Mechoulam Isolation and structure of a brain constituent that binds to the cannabinoid receptor. Devane W. A., Hanuš L., Breuer A., Pertwee R. G., Stevenson L. A., Griffin G., Gibson D., Mandelbaum A., Etinger A., Mechoulam R. Science 258, 1946-1949 (1992) 1 2 3 6 7 8 9 10 1. Inhibition of the specific binding of a radiolabelled cannabinoid probe to the synaptosomal membrain and work on isolation of anandamide (William A. Devane) 2. First isolation of anandamide from porcine brain (Lumír O. Hanuš, Tuesday, March 24, 1992) 3. Inhibition of the electrically evoked vas deferens twitch response (Roger Pertwee) 4. Inhibition of the electrically evoked vas deferens twitch response (Lesley A. Stevenson ) 5. Inhibition of the electrically evoked vas deferens twitch response (Graeme Griffin) 6. NMR spektrum of anandamide (Dan Gibson) 7. First high resolution mass spectrum (Asher Mandelbaum) 8. First high resolution mass spectrum (Alex Etinger) 9. Final structure of anandamide (Raphael Mechoulam, July 13, 1992) 10. First synthesis of anandamide (Aviva Breuer, August 20, 1992) Asher Gopher Israel Ringel First mass spectrum of anandamide (May 13, 1992) First NMR spectrum of anandamide and TMS-anandamide (May 14, 1992) האוניברסיטה העברית בירושלים HEBREW UNIVERSITY OF JERUSALEM Ester Fride The first in vivo examination of anandamide. After i.p. administration in mice caused lowering of activity in an immobility and in an open field test, and produced hypothermia and analgesia (these effects parallel those caused by psychotropic cannabinoids). E. Fride, R. Mechoulam: Pharmacological activity of the cannabinoid receptor agonist, anandamide, a brain constituent. Eur. J. Pharmacol. 231, 313-314(1993) O C OH N OH H O C5H11 D9-tetrahydrocannabinol Anandamide CB1 Lumir Hanuš,*, Asher Gopher,‡ Shlomo Almog‡, and Raphael Mechoulam*, RAPHAEL MECHOULAM, * SHIMON BEN-SHABAT,* LUMIR HANUŠ,* MOSHE LIGUMSKY,* NORBERT E. KAMINSKI, ‡ ANTHONY R. SCHATZ,§ ASHER GOPHER,॥ SHLOMO ALMOG,॥ BILLY R. MARTIN,§ DAVID R. COMPTON,§ ROGER G. PERTWEE,¶ GRAEME GRIFFIN,¶ MICHAEL BAYEWITCH,** JACOB BARG** and ZVI VOGEL** 2-arachidonoyl glycerol (2-AG) Presence of 2-AG in mouse bone May 2004 5 samples 70.4 mg ____________ June 2004 30 samples (femora and tibias) 0.6225 g 2-AG in bone Bone Remodeling Osteoclasts have the function A proper balance between to disrupt the old bone tissue osteoclasts and osteoblasts is the basis for a normal bone density OSTEOCLAST OSTEOBLAST A hyperactivity of osteoclasts is a one Osteoblasts have the function of the causes of the to produce the new bone tissue onset osteoporosis Medical illustration of the function of osteoblasts and osteoclasts in the regeneration of bone mass Anandamide reversed the inhibitory effect of AM251 on osteoclast formation 300 250 200 # 150 100 Osteoclasts per well per Osteoclasts ** 50 0 Vehicle AM251 AM251 + anandamide Nat. Med. 11, 774 (2005) Government statistics show that in Greece hip fractures are significantly lower than in North America and Northern Europe. Trichopoulou et al., have provided evidence that "The positive association between bone mineral density (BMD) and monounsaturated fat, which in the Greek diet is derived mostly from olive oil, is fairly strong, statistically significant" (Preventive Medicine 26, 395-400, 1997) Levels of bioactive lipids in cooking oils: olive oil is the richest source of oleoyl serine J Basic Clin Physiol Pharmacol. 27(3): 247–252 (2016) O O COOH H H C OH C OH N N arachidonoyl ethanolamide (anandamide) arachidonoyl serine COOH O H O H C OH C OH N N oleoyl ethanolamide oleoyl serine (HU-639) N-Oleoyl-L-serine is an endogenous lipid that has been reported to stimulate bone formation and to inhibit bone resorption. Oleoyl Serine Rescues Ovariectomy-induced Bone Loss 5 p= 0.065 (%) 4 p= 0.017 3 volume density volume - 2 1 trabecular bone trabecular