Research Journal of Pharmacy and Technology

Open Access Open Access  Restricted Access Subscription Access
Open Access Open Access Open Access  Restricted Access Restricted Access Subscription Access

Non-targeted Screening with Lc-hrms and In-silico Study on Diabetic Activity of Ethyl Acetate Extract of Sanrego (Lunasia Amara Blanco)


Affiliations
1 Doctoral Program of Medical Science, Faculty of Medicine, Universitas Brawijaya, Malang,, Indonesia
2 Department of Microbiology, Faculty of Medicine, Universitas Brawijaya, Malang,, Indonesia
3 Departmen of Biology, Faculty of Mathematics and Science Universitas Brawijaya, Malang,, Indonesia
4 Department of Pharmacy, Faculty of Medicine, Universitas Brawijaya, Malang,, Indonesia
     

   Subscribe/Renew Journal


Indonesian have long empirical use of the Sanrego plant (Lunasia amara Blanco) as antidiabetic, but the active compounds of Sanrego that acts as antidiabetic is not yet known. This study aimed to know the active compound from the ethyl acetate extract (EEA) of Sanrego stems and leaves and predict its ability as an anti- diabetic by in-silico. The dried leaves and stems of Sanrego were grounded into powder and extracted using ethyl acetate. The active compounds were detected using thin-layer chromatography (TLC) and Liquid chromatography high-resolution mass spectrometry (LC-HRMS). Anti-diabetic activity was predicted by molecular docking approach compared to acarbose and vildagliptin. The TLC results showed that Sanrego EEA contained alkaloid and flavonoid compounds include scopoletin. The LC-HRMS results showed 11 active compounds in EEA and all of them had anti-diabetic activity. The detected main compounds were hesperidin, scopoletin, tangeritin, and trigonelline. Based on the results of molecular docking, the four compounds showed anti-diabetic activity through α-glucosidase inhibition and dipeptidyl peptides- 4 (DPP-4) inhibition. Hesperidin has the highest energy affinity as an α-glucosidase inhibitor (-7.4) and DPP4 inhibitor (-9.8), followed by tangeritin, scopoletin, and trigonelline. This study concluded that the EEA of Sanrego contains hesperidin, tangeritin, scopoletin, and trigonelline which has anti-diabetic activity through α-glucosidase inhibition and DPP4 inhibition.

Keywords

Sanrego, α-Glucosidase, LC-HRMS, Hesperidin, Scopoletin.
Subscription Login to verify subscription
User
Notifications
Font Size


  • Adil M. P Ghosh. SK Venkata. Raygude K. Gaba D. Kandhare AD et al. Effect of Anti-Diabetic Drugs On Risk of Fracture In Type 2 Diabetes Mellitus Patients: A Network Meta-Analytic Synthesis of Randomized Controlled Trials of Thiazolidinediones. in ISPOR 20th Annual European Congress 2017. Glasgow, Scotland: Elsevier Inc. doi: https://doi.org/10.1016/j.jval.2017.08.724.

  • Khan MY. Poonam G, Bipin B, Vikas KV. A Review on Diabetes and Its Management. Asian J. Pharm. Res. 2013; 3(1):27-32. Available on: https://asianjpr.com/AbstractView.aspx?PID=2013-3-1-6

  • Yin Z. Zhang W. Feng F. Zhang Y. Kang W. α-Glucosidase Inhibitors Isolated From Medicinal Plants. Food Science and Human Wellness. 2014; 3:137-174. doi.org/10.1016/j.fshw.2014.11.003.

  • Mohapatra S. Prasad A. Haque F. Ray S. De B. Ray SS. In Silico Investigation of Black Tea Components on α -Amylase, α -Glucosidase and Lipase. J Appl Pharm Sci. 2015; 5(12):042-046. doi.org/10.7324/JAPS.2015.501207.

  • Purnomo Y. Soeatmaji DW. Sumitro SB. Widodo MA. Anti-diabetic Potential of Urena lobata Leaf Extract Through Inhibition of Dipeptidyl Peptidase IV Activity. Asian Pac J Trop Biomed. 2015; 5(8):645-649. doi.org/10.1016/j.apjtb.2015.05.014.

  • Subramanian R. Asmawi MZ. Sadikun A. In Vitro Alpha-Glucosidase and Alpha-Amylase Enzyme Inhibitory Effects of Andrographis paniculata Extract and Andrographolide. Acta Biochim Pol. 2008; 55(2):391-398.doi.org/10.18388/abp.2008_3087.

  • Zhang AJ. Rimando AM. Mizuno CS. Mathews ST. α-Glucosidase Inhibitory Effect of Resveratrol and Piceatannol. J Nutr Biochem. 2017; 47:86-93. doi.org/10.1016/j.jnutbio.2017.05.008

  • Wellington SJ., Amélio FG., Luiz GK. Dipeptidyl Peptidase 4: A New Link between Diabetes Mellitus and Atherosclerosis?. BioMed Research International. 2015; vol. 2015: 1-10.https://doi.org/10.1155/2015/816164

  • Tabopda TKT. Ngoupayo J. Awoussong PK. Mitaine-offer AC. Ali MS. Ngadjui BT. Triprenylated Flavonoids From Dorstenia psilurus and Their Alpha-Glucosidase Inhibition Properties. J Nat Pro. 2008; 71(12):2068-2072.doi.org/10.1021/np800509u.

  • Zafar M. Khan H. Rauf A. Khan A. Lodhi MA. In Silico Study of Alkaloids as α-Glucosidase Inhibitors: Hope For The Discovery of Efffective Lead Compounds. Front Endocrinol (Lausanne). 2016; 7:1-17. doi.org/10.3389/fendo.2016.00153.

  • Zhenzhen X. Wang G. Wang J. Chen M. Peng Y. Li Y et al. Synthesis, Biological Evaluation, and Molecular Docking Studies of Novel Isatin-thiazole Derivatives as α-Glucosidase Inhibitors. Molecules. 2017; 22(4):1-11. doi.org/10.3390/molecules22040659.

  • Gong Z. Peng Y. Qiu J. Cao A. Wang G. Peng Z. Synthesis, In Vitro α-Glucosidase Inhibitory Activity and Molecular Docking Studies of Novel Denzothiazole-triazole Derivatives. Molecules. 2017; 22(9):1-10.doi.org/10.3390/molecules22091555.

  • Spengler M, Schmitz H, Landen H. Evaluation of the efficacy and tolerability of acarbose in patients with diabetes mellitus. Clin Drug Invest. 2005;25:651–659. doi: 10.2165/00044011-200525100-00004.

  • Preethi J. Herbal Medicine for Diabetes Mellitus: A Review. Asian J. Pharm. Res. 2013; 3(2): 57-70. Available on: https://asianjpr.com/AbstractView.aspx?PID=2013-3-2-2

  • Macharla SP, , Venkateshwarlu G, Ravinder NA. Antidiabetic Activity of Bambusa arundinaceae Root Extracts on Alloxan Induced Diabetic Rats. Asian J. Res. Pharm. Sci. 2012; 2(2):73-75. Available on: https://ajpsonline.com/AbstractView.aspx?PID=2012-2-2-9

  • Saha D. Design and Potential of Mass Spectrometry. Asian J. Pharm. Ana. 2011; 1(1): 01-02. Available on: https://ajpaonline.com/AbstractView.aspx?PID=2011-1-1-1

  • Babu GR, J. Rao JS, Kumar KS, P. Reddy PJ. Stability Indicating Liquid Chromatographic Method for Aripiprazole. Asian J. Pharm. Ana. 2011;1(1):03-07. Available on: https://ajpaonline.com/AbstractView.aspx?PID=2011-1-1-2

  • Gupta A, Yadav JS, Rawat S , Mayuri G. Method Development and Hydrolytic Degradation Study of Doxofylline by RP-HPLC and LC-MS/MS. Asian J. Pharm. Ana. 2011; 1(1): 14-18. Available on: https://ajpaonline.com/AbstractView.aspx?PID=2011-1-1-5

  • Saha D, Tamrakar A. Protein Mass Spectrometry: Novel Approaches in Pharmaceutical Biotechnology. Asian J. Pharm. Ana. 2011; 1(2):25-26. Available on: https://ajpaonline.com/AbstractView.aspx?PID=2011-1-2-1

  • Pattabiraman K, Muthukumaran P. Antidiabetic and Antioxidant Activity of Morinda tinctoria roxb Fruits Extract in Streptozotocin-Induced Diabetic Rats. Asian J. Pharm. Tech. 2011; 1(2): 34-39 Available on: https://ajptonline.com/AbstractView.aspx?PID=2011-1-2-3

  • Chigozie IJ, Ikewuchi CC. Hypoglycemic, hypocholesterolemic, anti-anemic and ocular-protective effects of an aqueous extract of the rhizomes of Sansevieria liberica Gérôme and Labroy (Agavaceae) on alloxan induced diabetic Wistar rats. Asian J. Pharm. Tech. 2011; 1(4):137-148. Available on: https://ajptonline.com/AbstractView.aspx?PID=2011-1-4-9

  • Zubair MS. Anam S. Lallo S. Cytotoxic Activity and Phytochemical Standardization of Lunasia amara Blanco Wood Extract. Asian Pac J Trop Biomed. 2016; 6(11):962-966. doi.org/10.1016/j.apjtb.2016.04.014.

  • Luthfi MJ. Kamalrudin A. Noor MM. Effects of Lunasia amara Blanco (Sanrego) on Male Fertility: A Preliminary Study on Sperm Proteomic Analysis. J Appl Pharm Scie. 2017; 7(08):085-090. doi.org/10.7324/JAPS.2017.7081.

  • Hasnaeni. Sudarsono. Nurrochmat A. Widyarini S. Identification of Active Anti-inflammatory Principles of Beta-Beta Wood (Lunasia amara Blanco) From Siawung Barru-South Sulawesi, Indonesia. Trop J Pharm Res. 2017; 16:161-164. doi.org/10.4314/tjpr.v16i1.21.

  • Takahashi N. Subehan. Kadota S. Tezuka Y. Mechanism-based CYP2D6 Inactivation by Acridone Alkaloids of Indonesian Medicinal Plant

  • Lunasia amara. Fitoterapia. 2012; 83(4): 774-779. doi.org/10.1016/j.fitote.2012.03.011.

  • Ratnadewi AAI. Wahyudi LD. Rochman J. Susilawati. Ari S. Tri A. Revealing Anti-Diabetic Potency of Medicinal Plants of Meru Betiri National Park, Jember – Indonesia. Arab J Chem. 2018; 13(1):1831-1836.doi.org/10.1016/j.arabjc.2018.01.017.

  • Zhang Y. Wang N. Wang W. Wang J. Zhu Z. Li X. Molecular Mechanisms of Novel Peptides From Silkworm Pupae That Inhibit α- Glucosidase. Peptides. 2016;76:45-50. doi: 10.1016/j.peptides.2015.12.004.

  • Karumanchi SK. Atmakuri LR. Mandava VB. Rajala S. Synthesis and Hypoglycemic and Anti-Inflammatory Activity Screening of Novel Substituted 5-[morpholino(phenyl)methyl]-thiazolidine-2,4-diones and Their Molecular Docking Studies. Turk J Pharm Sci. 2019; 16(4):380- 390. doi.org/10.4274/tjps.galenos.2018.82612.

  • Hari S. In Silico Molecular Docking and ADME/T Analysis of Plant Compounds Against IL17A and IL18 Targets in Gouty Arthritis. J Appl Pharm Sci. 2019; 9(07):018-026. doi.org/10.7324/JAPS.2019.90703.

  • Peasari JR. Motamarry SS. Varma KS. Anita P. Potti RB. Chromatographic Analysis of Phytochemicals in Costus igneus and Computational Studies of Flavonoids. Informatics in Medicine Unlocked. 2018; 13:34-39.doi.org/10.1016/j.imu.2018.10.004.

  • Vo THN. Tran N. Nguyen D. Le L. An In Silico Study on Antidiabetic Activity of Bioactive Compounds in Euphorbia thymifolia Linn. Springerplus. 2016; 5(1):1-13. doi.org/10.1186/s40064-016-2631-5.

  • Ernawati T. Mun'Im A. Hanafi M. Yanuar A. In Silico Evaluation of Molecular Interactions Between Known α-Glucosidase Inhibitors and Homologous α-Glucosidase Enzymes From Saccharomyces cerevisiae, Rattus norvegicus, and GANC-human. Thai J Pharm Sci. 2018; 42(1):14-19. http://www.tjps.pharm.chula.ac.th/ojs/index.php/tjps/article/view/425

  • Adriani. Noorhamdani. Winarsih S. Ardyati T. Molecular Docking Study From Lunacridine, Scopoletin and Skimmianine as Antidiabetes Through α-Glucosidase Inhibitor, in The 1st International Seminar on Smart Molecule of Natural Resources (ISSMART). 2019, IOP Publishing Ltd: Swiss Bell Hotel Malang, Indonesia.1-7. doi.org/10.1088/1742-6596/1374/1/012026.

  • Macabeo APG. Alicia MA. Plant Review Chemical and Phytomedicinal Investigations in Lunasia amara. Phcog Rev. 2008; 2(4):317-325. https://www.researchgate.net/publication/236597615_Chemical_and_phytomedicinal_investigations_in_Lunasia_amara

  • Safitri A. Fatchiyah F. Sari DR. Roosdiana A. Phytochemical Screening, In Vitro Anti-oxidant Activity, and In silico Anti-diabetic Activity of Aqueous Extracts of Ruellia tuberosa L. J Appl Pharm Sci. 2020; 10(03):101-108. doi.org/10.7324/JAPS.2020.103013.

  • Deepak B, Jagtap RS, Kanase KG, Sonawame SA, Undale VR, Bhosale AV. Chemo informatics: Newer Approach for Drug Development. Asian J. Research Chem. 2009; 2(1): 01-07. Available on: https://ajrconline.org/AbstractView.aspx?PID=2009-2-1-1

  • Lin X. Li X. and Lin X. A Review on Applications of Computational Methods in Drug Screening and Design. Molecules. 2020; 25(3):1-17. doi.org/10.3390/molecules25061375.

  • Srinivasan S. Sadasivan SK. Exploring Docking and Aerobic-Microaerophilic Biodegradation of Textile Azo Dye by Bacterial Systems. J Water Process Eng. 2018; 22:180-191. doi.org/10.1016/j.jwpe.2018.02.004.

  • Arthur DE. Molecular Docking Studies of Some Topoisomerase II Inhibitors: Implications in Designing of Novel Anticancer Drugs. Radiol Infec Dis. 2019; 6(2):180-191. doi.org/10.1016/j.jrid.2019.06.003.

  • Jung UJ. Lee M. Jeong K. Choi M. The Hypoglycemic Effects of Hesperidin and Naringin are Partly Mediated by Hepatic Glucose- regulating Enzymes in C57BL/KsJ-db/db Mice. J Nutr. 2004; 134(10):2499-2503. doi.org/10.1093/jn/134.10.2499.

  • Akiyama S. Katsumata S. Suzuki K. Ishimi Y. Wu J. Uehara M. Dietary Hesperidin Exerts Hypoglycemic and Hypolipidemic Effects in Streptozotocin-induced Marginal Type 1 Diabetic Rats. J Clin Biochem Nutr. 2010; 46(1):87-92.doi.org/10.3164/jcbn.09-82.

  • Agrawal YO. Sharma PK. Shrivastapa B. Ojha S. Upadhya SM. Arya DS et al. Hesperidin Produces Cardioprotective Activity Via PPAR-γ Pathway in Ischemic Heart Disease Model in Diabetic Rats. PLoS One. 2014; 9(11):1-13. doi.org/10.1371/journal.pone.0111212.

  • Elshazly SM, El Motteleb DMA, and Ibrahim IAAE-H. Hesperidin Protects Against Stress Induced Gastric Ulcer Through Regulation of Peroxisome Proliferator Activator Receptor Gamma in Diabetic Rats. Chem Biol Interac. 2018; 291:153-161. doi.org/10.1016/j.cbi.2018.06.027.

  • Fitriawan L. Ariastuti R. Tjandrawinata RR. Nugroho AE. Pramono S. Antidiabetic Effect of Combination of Fractionated-Extracts of Andrographis paniculata and Centella asiatica: In vitro Study. Asian Pac J Trop Biomed. 2018; 8(11):527-532. doi.org/10.4103/2221- 1691.245957.

  • Dokumacioglu E. Iskender H. Musmul A. Effect of Hesperidin Treatment on α-Klotho/FGF-23 Pathway in Rats With Experimentally- Induced Diabetes. Biomed Pharmacother. 2018; 109:1206-1210. doi.org/10.1016/j.biopha.2018.10.192.

  • Zheng M, Shengmin L., Jianrong X., Enhanced Antioxidant, Anti-Inflammatory And α-Glucosidase Inhibitory Activities Of Citrus Hesperidin By Acid-Catalyzed Hydrolysis. Food Chemistry. 2021; Vol. 336: 1-9. doi. https://doi.org/10.1016/j.foodchem.2020.127539

  • Sahnoun M., Sahar, T., Samir B., Citrus Flavonoids Collectively Dominate The α-amylase and α-glucosidase Inhibitions. Biologia, 2017: 72(7): 764-773. DOI: 10.1515/biolog-2017-009

  • Ashrafizadeh M. Zahra A. Reza M. Elham GA. Tangeretin: A Mechanistic Review of Its Pharmacological and Therapeutic Effects. J Basic Clin Physiol Pharmacol. 2020; 31(4):1-7. doi.org/10.1515/jbcpp-2019-0191.

  • Kenji O. Horike N. Suzuki T. Hirano T. Polymethoxyflavonoids Tangeretin and Nobiletin Increase Glucose Uptake in Murine Adipocytes. Phytother Res. 2013; 27(2):312-316. doi.org/10.1002/ptr.4730.

  • Sundaram R. Shanthi P. and Sachdanandam P. Tangeretin, A Polymethoxylated Flavone, Modulates Lipid Homeostasis and Decreases Oxidative Stress by Inhibiting NF-κB Activation and Proinflammatory Cytokines in Cardiac Tissue of Streptozotocin-induced Diabetic Rats. J Func Foods. 2015; 16:315-333. doi.org/10.1016/j.jff.2015.03.024.

  • Cho J. Lee K. Park S. Jeong W. Isolation and Identification of α-Glucosidase Inhibitors from the Stem Bark of the Nutgall T ree ( Rhus javanica Linné). J Korean Soc Appl Biol Chem. 2013; 56:547-552. doi.org/10.1007/s13765-013-3140-7.

  • Jang JH. Park JEP. and Han JS. Scopoletin Inhibits α-Glucosidase In Vitro and Alleviates Postprandial Hyperglycemia In Mice With Diabetes. Eur J Pharmacol. 2018; 834:152-157. doi.org/10.1016/j.ejphar.2018.07.032.

  • Chang CC. Ho SL. and Lee SS. Acylated Glucosylflavones as α-Glucosidase Inhibitors From Tinospora crispa Leaf. Bioorg Med Chem. 2015; 23(13):3388-3396. doi.org/10.1016/j.bmc.2015.04.053.

  • Masitah K. Fazilah T. Nor SR, Saravanan D. Khamsah SM. Sasidharan S et al. In Vitro α-Amylase and α-Glucosidase Inhibition and Increased Glucose Uptake of Morinda citrifolia fruit and scopoletin. Research J. Pharm. and Tech. 2015; 8(2):189-193. https://rjptonline.org/AbstractView.aspx?PID=2015-8-2-6

  • Kalpana K. Priyadarshini E. Sreeja S. Jagan K. Anuradha CV. Scopoletin Intervention in Pancreatic Endoplasmic Reticulum Stress Induced by Lipotoxicity. Cell Stress Chaperones. 2018; 23(5):857-869.doi.org/10.1007/s12192-018-0893-2.

  • Jang JH. Park JE. Han JS. Scopoletin Increases Glucose Uptake Through Activation of PI3K and AMPK Signaling Pathway and Improves Insulin Sensitivity in 3T3-L1 Cells. Nutrition Research. 2020; 74:52-61.doi.org/10.1016/j.nutres.2019.12.003.

  • Subramanian SP. Prasath GS. Antidiabetic and Antidyslipidemic Nature of Trigonelline, A Major Alkaloid of Fenugreek Seeds Studied in High-fat-fed and Low-dose Streptozotocin-induced Experimental Diabetic Rats. Biomed Prev Nutr. 2014; 4(4):475-480. doi.org/10.1016/j.bionut.2014.07.001.

  • Li Y. Li Q. Wang C. Lou Z. Li Q. Trigonelline Reduced Diabetic Nephropathy and Insulin Resistance in Type 2 Diabetic Rats Through Peroxisome Proliferator‑Activated Receptor‑γ. Experimental and Therapeutic Medicine. 2019; 18(2):1331-1337. doi.org/10.3892/etm.2019.76

  • Gupta A., Glenn AJ., Jogn Rb., Herbert FJ., David SN., Christian KN., Hayder AA., Citrus Bioflavonoid Dipeptidyl Peptidase-4 Inhibition Compared With Gliptin Antidiabetic Medications. Biochemical and Biophysical Research Communications. 2018: 1-5. Doi. 10.1016/j.bbrc.2018.04.156

  • Hamden K., Amel B., Zahra A., Abdelfattah E., Experimental Diabetes Treated With Trigonelline: Effect on Key Enzymes Related to Diabetes and Hypertension, β-Cell and Liver Function. Mol Cell Biochem. 2013; 381(1-2): 85-94. doi: 10.1007/s11010-013-1690-y


Abstract Views: 107

PDF Views: 0




  • Non-targeted Screening with Lc-hrms and In-silico Study on Diabetic Activity of Ethyl Acetate Extract of Sanrego (Lunasia Amara Blanco)

Abstract Views: 107  |  PDF Views: 0

Authors

Adriani Adriani
Doctoral Program of Medical Science, Faculty of Medicine, Universitas Brawijaya, Malang,, Indonesia
Noorhamdani Noorhamdani
Department of Microbiology, Faculty of Medicine, Universitas Brawijaya, Malang,, Indonesia
Tri Ardyati
Departmen of Biology, Faculty of Mathematics and Science Universitas Brawijaya, Malang,, Indonesia
Sri Winarsih
Department of Pharmacy, Faculty of Medicine, Universitas Brawijaya, Malang,, Indonesia

Abstract


Indonesian have long empirical use of the Sanrego plant (Lunasia amara Blanco) as antidiabetic, but the active compounds of Sanrego that acts as antidiabetic is not yet known. This study aimed to know the active compound from the ethyl acetate extract (EEA) of Sanrego stems and leaves and predict its ability as an anti- diabetic by in-silico. The dried leaves and stems of Sanrego were grounded into powder and extracted using ethyl acetate. The active compounds were detected using thin-layer chromatography (TLC) and Liquid chromatography high-resolution mass spectrometry (LC-HRMS). Anti-diabetic activity was predicted by molecular docking approach compared to acarbose and vildagliptin. The TLC results showed that Sanrego EEA contained alkaloid and flavonoid compounds include scopoletin. The LC-HRMS results showed 11 active compounds in EEA and all of them had anti-diabetic activity. The detected main compounds were hesperidin, scopoletin, tangeritin, and trigonelline. Based on the results of molecular docking, the four compounds showed anti-diabetic activity through α-glucosidase inhibition and dipeptidyl peptides- 4 (DPP-4) inhibition. Hesperidin has the highest energy affinity as an α-glucosidase inhibitor (-7.4) and DPP4 inhibitor (-9.8), followed by tangeritin, scopoletin, and trigonelline. This study concluded that the EEA of Sanrego contains hesperidin, tangeritin, scopoletin, and trigonelline which has anti-diabetic activity through α-glucosidase inhibition and DPP4 inhibition.

Keywords


Sanrego, α-Glucosidase, LC-HRMS, Hesperidin, Scopoletin.

References