Cannabis Sativa: A Remedy for Convulsion by Inhibition of GABAA Receptor and Significantly Delayed Onset of Seizure Latency and Death - An Experimental Validation and Molecular Docking

Ben Enoluomen Ehigiator¹, SamuelKelechi Mobisson², Iheanyichukwu Wopara³*, Chinonye Cynthia Chibuife¹, Uwaezuoke Chukwuekele Awarajih³, Harrison Ogheneochuko Eruotor³

Abstract

This study aimed to investigate the anticonvulsant potentials of Cannabis sativa and its possible mechanism using the in vivo and in-silico models. A total of 20 male albino mice was used for this study and were divided randomly into five groups. Group 1 as negative control receiving 0.2ml 0f 10% ethanol only, and groups 2 and 3, received 200mg/kg/ body weight low dose of C. Sativa extract and 400mg/kg bw low dose of C. sativa extract, while groups 4 received 50 mg/kg body weight of phenobarbitone and group 5 received 50 mg/kg body weight of phenobarbitone+ 200 mg/kg body weight of the extract. Administration of C. Sativa extract was done via orogastric feeding; GABA-ergicseizure, was induced by administration of 80 mg/kg/bodyweight of Pentylenetetrazole (PTZ) via the intra-peritoneal route to all the groups. This was done about thirty minutes after the administration of treatment agents.  Each animal was observed individually for one hour to determine the onset of muscular jerks and tonic hind limb extensions and death.  Animal study results revealed that C. sativa may likely possess’ anticonvulsant potentials as It is significantly seizure latency and death in PTZ treated mice. For the in silico studies, about 49 compounds of C. sativa were obtained from the PubChem library and were docked on various targets site of proteins; Glutaminase, 4-Aminobtyrate Aminotransferase, Perioxisome proliferator Activated receptor (PPAR), Succinic semialdehyde dehydrogenase, Gama amino butyric acid –A receptor (GABA-A), Histone deacetylases (HDAC), Phosphoribosyl pyrophosphate amido transferase and human GAFT-1. Furthermore, the in-silico study revealed that compounds like Cannabisin A and D showed anticonvulsant potential activation of GABA.

Key Words: Convulsions, Anticonvulsant, Cannabis Sativa, in-silico studies, Glutaminase  

References

Anigilaje E, Anigilaje OO. (2012). Childhood convulsion: inquiry about the concerns and home management among mothers in tegbesun, a periurban community in Ilorin, Nigeria. Paediatrics (4):209609.

Berman HM, Westbrook, J, Feng Z, Gillialand G, Bhat TN, Weissig H, Shindyalov IN, Bourne PE. (2000). The Protein data bank.Nucleic Acids Research 28(1):235-242.

BrenneisenR. (2007). “Chemistry and analysis of phytocannabinoids and other cannabis constituents,” in Marijuana and the Cannabinoids Forensic Science and Medicine, ed. M. ElSohly (New York, NY: Humana Press), 17–49. doi: 10.1007/978-1-59259-947-9.

Bromfield  EB. (2006). Basic mechanisms underlying seizures and epilepsy -An introduction to epilepsy.American Epilepsy Society: B (slide 2)

Brooks, WH, Daniel KG, Sung SS, Guida, WC. (2008). Computational validation of the importance of absolute stereochemistry in virtual screening.Journal of Chemical Information and Modeling; 48: 639–45.

Chou KC. (2004). Structural bioinformatics and its impact to biomedical science.Current Medicinal Chemistry.11(16):2105–2134.

Edward HR. (2000). The ILAE/IBE/WHO global campaign against epilepsy: bringing epilepsy “out of the shadows”. Epilepsy & Behavior, 1(4), S3-S8.

Ekeh BC. (2015). The knowledge, attitude and perception towards Epilepsy amongst medical students in Uyo, southern Nigeria. Advances in Medicine :876135.

Elsohly M, El-Feraly MA. (2005). Chemical constituents of marijuana- the complex mixture of natural cannabinoids. The Haworth Press, Inc., Binghamton, New York, pages 27-36

Emilio P. (2017). Cannabinoids in the treatment of epilepsy: hard evidence at last. Journal of Epilepsy Research 7(2); 61-76.

EubanksLM, Rogers C J, Beuscher AE,  Koob GF, Olson AJ, Dickerson TJ, Janda  KD. (2006). "A Molecular Link Between the Active Component of Marijuana and Alzheimer's Disease Pathology". Molecular Pharmaceutics3 (6): 773–7.

Guyton AC, Hall JE. (2012). Textbook of medical physiology (10^thed.). Philadelphia: W. B. Saunders, 386 – 387.

Hendricks H, Malingre TM, Batterman S, Bos R. (1977). Alkanes of the essential oil of cannabis sativa. Available at https://www.semanticscholar.org/paper/alkanes-of-the-essential-oil-of-cannabis-sativa accessed [14^th November, 2020].

Hetenyi C, Vander-Spoel  D. (2002).” Efficient docking of peptides to proteins without prior knowledge of the binding site”.Protein Science.;11:1729-1737.

Huntsman R, Tang-wai R, ZentenoJ. (2019).Cannabis for pediatric and adult epilepsy.Available on https://www.intechopen.com doi:10.5772/intechopen.85719.

HuzarT. (2019). Everything you need to know about convulsions. Available at https://www.medicalnewstoday.com/articles/324330.

Katja EB. (2020). Cognitive and Psychological Side Effects of Antiepileptic Drugs-Update on classification, etiologies, instrumental diagnosis and treatment.Intechopen, doi:10.5772/intechopen.94308.

Kvamme E, Torgner I, Roberg B. (2001). Kinetics and localization of brain phosphate activated glutaminase. Journal of Neuroscience Research; 66:951-958.

Lipinski CA. (2001). Avoiding investment in doomed drugs.Current drug discovery, 1; 17-19.

Mediavilla V, Steinmann S. (1997). Essential oil of cannabis sativa L. strains.Journal international hemp association 4:80-82

Mobisson SK, Obembe AO, Ukoh IE,  Duru, GO. (2018). The Role of Hypothalamic Ptuitary Gonadal Axis in Aqueous Extract of Cannabis Sativa Induced Male Reproductive Dysfunction of Albino Wistar Rats. European Journal of Pharmaceutical and Medical Research,5(1), 71-78.

Morris GM, Huey R, Lindstrom W, Sanner MF, Belew RK,  Goodsell DS. (2009).  “AutoDock 4 and Auto Dock Tools 4: Automated docking with selective receptor flexibility”. Journal of Computational Chemistry.30:2785-2791.

Novak J, Zitter I-Eglseer K, Deens S.G, Franz C.M. (2001). Essential oils of different cultivars of cannabis sativa L. and other antimicrobial activity; flavour and fragrance journal 16; 259-262.

Perucca E. (2017). Cannabinoids in the Treatment of Epilepsy: Hard Evidence at Last?. Journal of epilepsy research, 7(2), 61–76.https://doi.org/10.14581/jer.17012.

Phillip L, Shukla, L, William, H, Jakobs C, Gibson K. (2011). Epilepsy in Succinic Semialdehyde Dehydrogenase Deficiency, a Disorder of GABA Metabolism.US National Library of Medicine 33(9); 796-805.

Puligheddu, M, Pillolla G, Melis M. (2013). PPAR-alpha agonists as novel antiepileptic drugs: Preclinical findings. PLoS ONE;8(5):64541.

Railton D. (2018). Marijuana: good or bad. Medical News Today.BMC Psychiatry, 11 (1), 25.Available at https://www.medicalnewstoday.com .

Russo EB. (2017). Cannabis and Epilepsy: An ancient treatment returns to the fore. Epilepsy behaviour, 70: 292-297.

Sastry GM, Adzhigrey M, Day T, Annabhimoju R, Sherman W. (2013). Protein and Ligand preparation: Parameters, protocols, and influence on virtual screening enrichments.Journal of Computer-Aided Molecular Design , 27(3),221-234.

Schauer G, King B, Bunnel R, Promoff G,  McAfee T. (2016). Toking, vaping, and eating for health or fun: marijuana use patterns in adults, U.S., American Journal of Preventive Medicine. 50(1):1-8.

Sidra Z, Deepark K, Muhammad TK, Pirthvi RG.  Fnu, K. (2018). Epilepsy and cannabis: A Literature Review. Neurology, 10(9): e3278.

Suraev AS, Todd L, Bowen MT, Alsop DJ, McGregor IS, Ireland C, Lintzeris N. (2017).  Australian nationwide survey on medicinal cannabis use for epilepsy: history of antiepileptic drug treatment predicts medicinal cannabis use. Epilepsy behaviour 70:334-334.