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Chlorogenic acid and gallic acid synergistically reduce hyperglycemia and hyperlipidemia in diabetic BALB/c mice
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  • Published: 17 March 2026

Chlorogenic acid and gallic acid synergistically reduce hyperglycemia and hyperlipidemia in diabetic BALB/c mice

  • Mudassir Hassan1,2,
  • Muhammad Ali3,
  • Javaria Altaf4 &
  • …
  • Faisal Saeed Awan  ORCID: orcid.org/0000-0003-1525-83731 

Scientific Reports , Article number:  (2026) Cite this article

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We are providing an unedited version of this manuscript to give early access to its findings. Before final publication, the manuscript will undergo further editing. Please note there may be errors present which affect the content, and all legal disclaimers apply.

Subjects

  • Diseases
  • Drug development
  • Drug discovery
  • Endocrine system and metabolic diseases
  • Endocrinology
  • Metabolic disorders
  • Translational research

Abstract

Given the multifactorial nature of diabetes and limitations of conventional monotherapies, combination therapies involving natural bioactive compounds offer a complementary strategy for more effective management. This study investigated the antidiabetic potential of chlorogenic acid (CGA) and gallic acid (GA), individually and in combination, using in silico, in vitro and in vivo approaches. Pharmacokinetic and molecular docking analyses were conducted to assess drug-likeness and target interactions. Antioxidant activity and α-amylase inhibition were evaluated in vitro, while therapeutic efficacy was assessed in streptozotocin-nicotinamide-induced (STZ-NA) diabetic mice. In silico analysis revealed favorable ADMET profiles and strong binding affinities of CGA and GA to key diabetic targets, including α-amylase, DPP4, PPARγ, TNF-α and IL-6, surpassing metformin. The combination showed enhanced antioxidant and α-amylase enzyme inhibitory activity compared to individual compounds. In vivo findings using an STZ-NA-induced type II diabetic mouse model demonstrated a significant reduction in blood glucose (p < 0.0001) and improvement in lipid profile upon oral supplementation. Both compounds showed a decline in gene expression of TNF-α, TGF-β and IL-6 while enhancing the level of PPARγ. Additionally, hepatic markers (ALT, AST and ALP) were significantly reduced (p < 0.05), suggesting hepatoprotective effects. Histological analysis revealed restoration of pancreatic and liver tissues. Notably, the combination of both compounds exhibited a greater antidiabetic effect than the individual compounds and the standard antidiabetic drug (metformin). Given their favorable pharmacokinetic profile, affinity for multiple antidiabetic targets and synergistic antidiabetic activities both in vitro and in vivo, CGA-GA combination may offer a safer alternative to current therapies. These findings warrant further investigation of this combination strategy for more effective diabetes management.

Data availability

All data supporting the findings of this study are included in this manuscript. Additional data are available from the corresponding author upon reasonable request at faisal.saeed@uaf.edu.pk.

Abbreviations

ADMET:

Absorption, distribution, metabolism, excretion and toxicity

ALP:

Alkaline phosphatase

ALT:

Alanine aminotransferase

AST:

Aspartate aminotransferase

DPP4:

Dipeptidyl peptidase-4

IL-6:

Interleukin-6

PPARγ:

Peroxisome proliferator-activated receptor gamma

TGF-β:

Transforming growth factor-beta

TNF-α:

Tumor necrosis factor-alpha

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Acknowledgements

We sincerely thank Ms. Shagufta Malik, Department of Biochemistry, University of Agriculture, Faisalabad, for her valuable inputs and generous help in proofreading the manuscript.

Funding

This research received no external funding.

Author information

Authors and Affiliations

  1. Centre of Agricultural Biochemistry and Biotechnology, University of Agriculture Faisalabad (UAF), Faisalabad, 38040, Pakistan

    Mudassir Hassan & Faisal Saeed Awan

  2. Graduate Institute of Biomedical Sciences, China Medical University, Taichung, Taiwan

    Mudassir Hassan

  3. Department of Biochemistry, Faculty of Sciences, University of Agriculture Faisalabad (UAF), Faisalabad, 38040, Pakistan

    Muhammad Ali

  4. Department of Zoology, Government College University, Faisalabad, Pakistan

    Javaria Altaf

Authors
  1. Mudassir Hassan
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  2. Muhammad Ali
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  3. Javaria Altaf
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  4. Faisal Saeed Awan
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Contributions

M.H.: Conceptualization, Methodology, Software, Validation, Investigation, Data curation, Writing – original draft, Writing – review & editing. F.S.A.: Conceptualization, Methodology, Validation, Formal analysis, Resources, Writing – review & editing, Visualization, Supervision. M.A.: Conceptualization, Methodology, Validation, Resources, Supervision. J.A.: Resources, Writing – review & editing.

Corresponding author

Correspondence to Faisal Saeed Awan.

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Competing interests

The authors declare no competing interests.

Institutional review board statement

All animal procedures were approved by the Institutional Biosafety/Bioethics Committee (IBC) at the University of Agriculture Faisalabad (Approval No. D.No 1957/ORIC, dated 06-05-2024) and conducted following the National Biosafety Guidelines 2005 (Amended 2024), the Punjab Animal Health Act, 2019 and Institutional Bioethics Protocols. The study complies with the ARRIVE guidelines.

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Hassan, M., Ali, M., Altaf, J. et al. Chlorogenic acid and gallic acid synergistically reduce hyperglycemia and hyperlipidemia in diabetic BALB/c mice. Sci Rep (2026). https://doi.org/10.1038/s41598-026-37188-5

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  • Received: 30 May 2025

  • Accepted: 20 January 2026

  • Published: 17 March 2026

  • DOI: https://doi.org/10.1038/s41598-026-37188-5

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Keywords

  • Combination therapy
  • Antioxidant activity
  • ADMET analysis
  • Molecular docking
  • Chlorogenic acid
  • Gallic acid
  • In vivo antidiabetic activity
  • Expression analysis
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