Phenolic phytochemistry, in vitro, in silico, in vivo, and mechanistic anti-inflammatory and antioxidant evaluations of Habenaria digitata

Hassan Hussain Almasoudi; Muhammad Saeed Jan; Mohammed H Nahari; Abdulfattah Yahya M Alhazmi; Abdulkarim S Binshaya; Osama Abdulaziz; Mater H Mahnashi; Muhammad Ibrar; Rehman Zafar; Abdul Sadiq

doi:10.3389/fphar.2024.1346526

Phenolic phytochemistry, in vitro, in silico, in vivo, and mechanistic anti-inflammatory and antioxidant evaluations of Habenaria digitata

Front Pharmacol. 2024 Feb 29:15:1346526. doi: 10.3389/fphar.2024.1346526. eCollection 2024.

Affiliations

¹ Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, Najran University, Najran, Saudi Arabia.
² Department of Pharmacy, Bacha Khan University, Charsadda, Pakistan.
³ Pharmaceutical Practices Department, College of Pharmacy, Umm Al-Qura University, Makkah, Saudi Arabia.
⁴ Department of Medical Laboratory Sciecnes, College of Applied Medical sciences, Prince Sattam bin Abdulaziz University, Alkharj, Saudi Arabia.
⁵ Clinical Laboratory Sciences Department, College of Applied Medical Sciences, Taif University, Taif, Saudi Arabia.
⁶ Department of Pharmaceutical Chemistry, Pharmacy School, Najran University, Najran, Saudi Arabia.
⁷ Department of Pharmaceutical Chemistry, Faculty of Pharmaceutical Sciences, Riphah International University, Islamabad, Pakistan.
⁸ Department of Pharmacy, Faculty of Biological Sciences, University of Malakand, Chakdara, Khyber Pakhtunkhwa, Pakistan.

Abstract

Excessive and imbalance of free radicals within the body lead to inflammation. The objective of the current research work was to explore the anti-inflammatory and antioxidant potential of the isolated compounds from Habenaria digitata. In this study, the isolated phenolic compounds were investigated for in vitro and in vivo anti-inflammatory potential along with the antioxidant enzyme. The anti-inflammatory and antioxidant potential of the phenolic compounds was assayed via various enzymes like COX-1/2, 5-LOX and ABTS, DPPH, and H₂O₂ free radical enzyme inhibitory assay. These compounds were also explored for their in vivo antioxidant activity like examining SOD, CAT, GSH-Px, and MDA levels in the brain, heart, and liver. The anti-inflammatory potential was evaluated using the carrageenan-induced pleurisy model in mice. On the basis of initial screening of isolated compounds, the most potent compound was further evaluated for the anti-inflammatory mechanism. Furthermore, the molecular docking study was also performed for the potent compound. The phenolic compounds were isolated and identified by GC-MS/NMR analysis by comparing its spectra to the library spectra. The isolated phenolic compounds from H. digitata were 5-methylpyrimidine-24,4-diol (1), 3,5-dihydroxy-6-methyl-2,3-dihydropyran-4-one (2), 2-isopropyl-5-methylphenol (3), 3-methoxy-4-vinylphenol (4), and 2,6-dimethoxy-4-vinylphenol (5). In in vitro antioxidant assay, the most potent compound was compound 1 having IC₅₀ values of 0.98, 0.90, and 5 μg/mL against ABTS, DPPH, and H₂O₂, respectively. Similarly, against COX1/2 and 5-LOX ,compound 1 was again the potent compound with IC₅₀ values of 42.76, 10.70, and 7.40 μg/mL. Based on the in vitro results, compound 1 was further evaluated for in vivo antioxidant and anti-inflammatory potential. Findings of the study suggest that H. digitata contains active compounds with potential anti-inflammatory and antioxidant effects. These compounds could be screened as drug candidates for pharmaceutical research, targeting conditions associated with oxidative stress and inflammatory conditions in medicinal chemistry and support their ethnomedicinal use for inflammation and oxidative stress.

Keywords: Habenaria digitata; anti-inflammatory; antioxidant; mechanism; molecular docking; phenolic.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. Authors would like to acknowledge the support of the Deputy for Research and Innovation Ministry of Education, Kingdom of Saudi Arabia, for this research through a grant (NU/IFC/02//MRC/-004) under the Institutional Funding Committee at Najran University, Kingdom of Saudi Arabia.