Integration of in silico, in vitro and ex vivo pharmacology to decode the anti-diabetic action of Ficus benghalensis L. bark



Traditionally, Ficus benghalensis L. is used to treat metabolic disorders and is also recorded in the Ayurvedic pharmacopeia of India. The present study aimed to evaluate the anti-diabetic property of hydroalcoholic extract/fraction(s) of F. benghalensis L. bark via in silico, in vitro, and ex vivo approach.


Enzyme inhibitory activity, glucose uptake in rat hemidiaphragm, and glucose permeability, and adsorption assays were performed using in vitro and ex vivo methods as applicable. Further, the PASS was used to identify the probable lead enzyme inhibitors. The presence of predicted enzyme inhibitors was confirmed via the LC-MS. Similarly, the docking of ligands with respective targets was performed using autodock4.0.


Flavonoids rich fraction possessed the highest α-amylase, and α-glucosidase inhibitory activity followed by maximum efficacy for glucose uptake in rat hemidiaphragm. Similarly, the hydroalcoholic extract showed the highest efficacy to inhibit glucose diffusion. Likewise, 3,4-dihydroxybenzoic acid was predicted for the highest pharmacological activity for α-amylase, ursolic acid for PTP1B, and apigenin for α-glucosidase inhibition respectively. The LC-MS analysis also identified the presence of the above hit molecules in the hydroalcoholic extract.


The analogs of 3,4-dihydroxybenzoic acid, apigenin, and ursolic acid could be the choice of lead hits as the α-amylase, α-glucosidase, and PTP1B inhibitors respectively. Additionally, the majority of secondary metabolites from the hydroalcoholic extract of F. benghalensis may be involved in enhancing the glucose uptake to support the process of glycogenesis.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Data availability

Data will be provided in case of a request.



area under the curves


Dinitrosalicylic acid


Effective concentration 50


Glucose transporter


Institutional Animal Ethics committee


Indian Council of Medical Research - National Institute of Traditional Medicine


Liquid chromatography-mass spectrometry


Molecular formula


Molecular weight


Pharmacological activity


Prediction of Activity Spectra for Substances


Pharmacological inactivity


4-Nitrophenyl-β-D- glucopyranoside


Protein Tyrosine Phosphatase 1B


Research Collaboratory for Structural Bioinformatics


Simplified molecular-input line-entry system


World Health Organization


  1. 1.

    Cerf ME. Beta cell dysfunction and insulin resistance. Front Endocrinol (Lausanne). 2013;4:37.

    Article  Google Scholar 

  2. 2.

    Lorenzati B, Zucco C, Miglietta S, Lamberti F, Bruno G. Oral hypoglycemic drugs: pathophysiological basis of their mechanism of action. Pharmaceuticals (Basel). 2010;3(9):3005–20.

    Article  CAS  Google Scholar 

  3. 3.

    Pan SY, Zhou SF, Gao SH, Yu ZL, Zhang SF, Tang MK, et al. New perspectives on how to discover drugs from herbal medicines: CAM’s outstanding contribution to modern therapeutics. Evid Based Complement Alternat Med. 2013;2013:627375.

    PubMed  PubMed Central  Google Scholar 

  4. 4.

    Khanal P, Patil BM. α-Glucosidase inhibitors from Duranta repens modulate p53 signaling pathway in diabetes mellitus. Adv Tradit Med (ADTM) 2020. Available at:

  5. 5.

    WHO. Diabetes mellitus. Report of a WHO expert committee. 1965. p. 1–44. Available from:;jsessionid=2C4D0847578CDB3E30D98B6B54F12241?sequence=1.

  6. 6.

    Government of India, Ministry of Health and Family Welfare, Department of AYUSH. The Ayurvedic Pharmacopoeia of India. Part- 1, Vol. 1, Page No. 119.

  7. 7.

    Rajdev K, Jain S, Mahendra CH, Bhattacharaya SK. Antinociceptive effect of Ficus bengalensis bark extract in experimental models of pain. Cureus. 2018;10(3):e2259.

    PubMed  PubMed Central  Google Scholar 

  8. 8.

    Patel DK, Prasad SK, Kumar R, Hemalatha S. An overview on antidiabetic medicinal plants having insulin mimetic property. Asian Pac J Trop Biomed. 2012;2(4):320–30.

    Article  CAS  Google Scholar 

  9. 9.

    de Souza PM, de Oliveira Magalhães P. Application ofmicrobial α-amylase in industry - a review. Braz JMicrobiol. 2010;41(4):850–61.

    Article  Google Scholar 

  10. 10.

    Telagari M, Hullatti KK. In-vitro α-amylase and α-glucosidase inhibitory activity of Adiantum caudatum Linn. And Celosia argentea Linn. Extracts and fractions. Indian J Pharmacol. 2015;47(4):425–9.

    Article  CAS  Google Scholar 

  11. 11.

    Wang C, Wang L, Yang Z. Role of protein tyrosine phosphatase 1B in the type 2 diabetes and obesity. Yi Chuan. 2004 Nov;26(6):941–6. Chinese.

  12. 12.

    Fernandez-Ruiz R, Vieira E, Garcia-Roves PM, Gomis R. Protein tyrosine phosphatase-1B modulates pancreatic β-cell mass. PLoS One. 2014 Feb 28;9(2):e90344. doi: PMID: 24587334; PMCID: PMC3938680.

  13. 13.

    Khanal P, Patil BM. Gene set enrichment analysis of alpha-glucosidase inhibitors from Ficus benghalensis. Asian Pac J Trop Biomed. 2019;9(6):263–70.

    Article  CAS  Google Scholar 

  14. 14.

    Khanal P, Patil BM. Gene ontology enrichment analysis of α-amylase inhibitors from Duranta repens in diabetes mellitus. J Diabetes Metab Disord. 2020.

  15. 15.

    Chattopadhyay RR, Sarkar SK, Ganguly S, Banerjee RN, Basu TK. Effect of leaves of Vinca rosea Linn, on glucose utilization and glycogen deposition by isolated rat hemidiaphragm. Indian J Physiol Pharmacol. 1992;36:137–8.

    PubMed  CAS  Google Scholar 

  16. 16.

    Ou S, Kwok K, Li Y, Fu L. In vitro study of possible role of dietary fiber in lowering postprandial serum glucose. J Agric Food Chem. 2001;49(2):1026–9.

    Article  CAS  Google Scholar 

  17. 17.

    Gallagher AM, Flatt PR, Duffy G, Abdel-Wahab YHA. The effects of traditional antidiabetic plants on in vitro glucose diffusion. Nutr Res. 2003;23(3):413–24.

    Article  CAS  Google Scholar 

  18. 18.

    Dixit P, Jain DK, Dumbwani J. Standardization of an ex vivo method for determination of intestinal permeability of drugs using everted rat intestine apparatus. J Pharmacol Toxicol Methods. 2012;65(1):13–7.

    Article  PubMed  CAS  Google Scholar 

  19. 19.

    Filimonov DA, Lagunin AA, Gloriozova TA, Rudik AV, Druzhilovskii DS, Pogodin PV, et al. Prediction of the biological activity spectra of organic compounds using the PASS online web resource. Chem Heterocycl Comp. 2014;50(3):444–57.

    Article  CAS  Google Scholar 

  20. 20.

    Halgren TA. Merck molecular force field. I. Basis, form, scope, parameterization, and performance of MMFF94. J Comput Chem. 1996;17(5–6):490–519.

    Article  CAS  Google Scholar 

  21. 21.

    Schwede T, Kopp J, Guex N, Peitsch MC. SWISS-MODEL: an automated protein homology-modeling server. Nucleic Acids Res. 2003;31(13):3381–5.

    Article  CAS  Google Scholar 

  22. 22.

    Morris GM, Huey R, Lindstrom W, Sanner MF, Belew RK, Goodsell DS, et al. AutoDock4 and AutoDockTools4: automated docking with selective receptor flexibility. J Comput Chem. 2009;30:2785–91.

    Article  CAS  Google Scholar 

  23. 23.

    Bakke J, Haj FG. Protein-tyrosine phosphatase 1B substrates and metabolic regulation. Semin Cell Dev Biol. 2015;37:58–65.

    Article  CAS  Google Scholar 

  24. 24.

    Panzhinskiy E, Ren J, Nair S. Protein tyrosine phosphatase 1B and insulin resistance: role of endoplasmic reticulum stress/reactive oxygen species/nuclear factor kappa B axis. PLoS One. 2013;8(10):e77228.

    Article  CAS  Google Scholar 

  25. 25.

    Liu T, Lin Y, Wen X, Jorissen RN, Gilson MK. BindingDB: a web-accessible database of experimentally determined protein-ligand binding affinities. Nucleic Acids Res. 2007;35:D198–201.

    Article  CAS  Google Scholar 

  26. 26.

    Saeidnia S, Ara L, Hajimehdipoor H, Read RW, Arshadi S, Nikan M. Chemical constituents of Swertia longifolia Boiss. With α-amylase inhibitory activity. Res Pharm Sci. 2016;11(1):23–32.

    PubMed  PubMed Central  Google Scholar 

  27. 27.

    Dehghan H, Salehi P, Amiri MS. Bioassay-guided purification of α-amylase, α-glucosidase inhibitors and DPPH radical scavengers from roots of Rheum turkestanicum. Ind Crop Prod. 2018;117(2018):303–9.

    Article  CAS  Google Scholar 

  28. 28.

    Tadera K, Minami Y, Takamatsu K, Matsuoka T. Inhibition of alpha-glucosidase and alpha-amylase by flavonoids. J Nutr Sci Vitaminol (Tokyo). 2006 Apr;52(2):149–53.

    Article  CAS  Google Scholar 

  29. 29.

    Proença C, Freitas M, Ribeiro D, Sousa JLC, Carvalho F, Silva AMS, et al. Inhibition of protein tyrosine phosphatase 1B by flavonoids: a structure - activity relationship study. Food Chem Toxicol. 2018 Jan;111:474–81.

    Article  PubMed  CAS  Google Scholar 

  30. 30.

    Zhang BW, Xing Y, Wen C, Yu XX, Sun WL, Xiu ZL, et al. Pentacyclic triterpenes as α-glucosidase and α-amylase inhibitors: structure-activity relationships and the synergism with acarbose. Bioorg Med Chem Lett. 2017;27(22):5065–70.

    Article  PubMed  CAS  Google Scholar 

  31. 31.

    Zhao BT, Nguyen DH, Le DD, Choi JS, Min BS, Woo MH. Protein tyrosine phosphatase 1B inhibitors from natural sources. Arch Pharm Res. 2018;41(2):130–61.

    Article  PubMed  CAS  Google Scholar 

Download references


The authors are thankful to Principal KLE College of Pharmacy Belagavi for providing necessary facilities to complete the work. Pukar Khanal is thankful to Ms. Taaza Duyu (Department of Pharmacology and Toxicology, KLE College of Pharmacy Belagavi) for her assistance during the enzyme inhibitory activity, Rohini S. Kavalapure (Department of Pharmaceutical Chemistry, KLE College of Pharmacy Belagavi) for interpreting LC-MS data, Dr. Manish Wanjari (Regional Ayurveda Research Institute for Drug Development Gwalior-474009, Madhya Pradesh, India) for his suggestion for glucose permeability assay and Dr. Yadu Nandan Dey for his suggestion for drafting this manuscript.


This work has not received any funds from national and international agencies.

Author information



Corresponding authors

Correspondence to Pukar Khanal or B. M. Patil.

Ethics declarations

Conflict of interest

There are no conflicts of interest to declare.

Ethical statement

Glucose uptake and permeability assayes were performed after receiving ethical clearance from Institutional animal ethical clearance (IAEC) at KLE College of Pharmacy, Belagavi (resolution no. KLECOP/CPCSEA-Reg, No.221/Po/Re/S/2000/CPCSEA, Res.28–12/10/2019).

Consent for publication

Not Applicable

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material


(DOCX 7.74 MB)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Khanal, P., Patil, B.M. Integration of in silico, in vitro and ex vivo pharmacology to decode the anti-diabetic action of Ficus benghalensis L. bark. J Diabetes Metab Disord (2020).

Download citation


  • Apigenin
  • Diabetes mellitus
  • Ficus benghalensis
  • Postprandial hyperglycemia