Sains Malaysiana 52(6)(2023): 1635-1648

http://doi.org/10.17576/jsm-2023-5206-03

 

In Vitro and In Silico Study on the Interaction between Apigenin, Kaempferol and 4-Hydroxybenzoic Acid in Xanthine Oxidase Inhibition

(Kajian Secara In Vitro dan In Silico pada Interaksi antara Apigenin, Kaempferol dan Asid 4-Hidroksibenzoik dalam Perencatan Xantina Oksidase)

 

CHIN YONG SIN1, LOH KHYE ER1,*, WEE SZE PING1 & ONG GHIM HOCK2

 

1Department of Bioscience, Faculty of Applied Sciences, Tunku Abdul Rahman University of Management and Technology, Jalan Genting Kelang, Setapak. 53300 Kuala Lumpur, Malaysia

2Faculty of Health and Life Sciences, INTI International University, Persiaran Perdana BBN, Putra Nilai, 71800 Nilai, Negeri Sembilan, Malaysia

 

Received: 17 August 2022/Accepted: 23 May 2023

 

Abstract

Xanthine oxidase (XO) is a biological enzyme that takes part in purine catabolism. It catalyses the conversion of hypoxanthine to xanthine and eventually xanthine to uric acid. The catabolism reaction increases the level of uric acid and subsequently leads to hyperuricemia. Allopurinol is a XO inhibitor that is used clinically to prevent purine catabolism. Although it is an effective XO inhibitor, it causes some side effects. Therefore, a more effective inhibitor with fewer side effects is in an urgent need. Phenolic compounds have been identified as effective XO inhibitors in many studies. In vitro and in silico study were conducted to investigate the interaction between apigenin, kaempferol and 4-hydroxybenzoic acid in XO inhibition. Apigenin was found to be the most effective XO inhibitor among the compounds tested with the best docking score of -8.2 kcal/mol as demonstrated in the molecular docking simulation which indicated its favourable interaction with XO enzyme. Additive interactions between compounds namely apigenin-kaempferol, apigenin-4-hydroxybenzoic acid and 4-hydroxybenzoic acid-kaempferol were demonstrated in both in vitro and in silico studies. The results showed that 4-hydroxybenzoic acid- apigenin (-7.4 kcal/mol) was the most stable ligands combination docked to XO. The multiple ligands docking simulation showed independent ligands bound to the XO active site at non-interfering regional location. In conclusion, the combination of these three compounds can be explored further for their additive interaction in XO inhibition, which could be beneficial in terms of the enhanced effectiveness and lower side effects when each is used at lower dose to give the same effect.

 

Keywords: Additive interaction; molecular docking; multiple ligands; phenolic compounds; xanthine oxidase inhibitor

 

Abstrak

Xantina oksidase (XO) ialah sejenis enzim biologi yang terlibat dalam metabolisme purin. Ia memangkinkan penukaran hipozantin kepada xantina dan akhirnya daripada xantina kepada asid urik. Tindak balas katabolisme meningkatkan tahap asid urik dan seterusnya membawa kepada hiperurisemia. Allopurinol adalah sejenis perencat XO yang digunakan secara klinikal untuk mencegah katabolisme purin. Walaupun ia adalah sejenis perencat XO yang berkesan, ia menyebabkan kesan sampingan. Oleh itu, perencat yang lebih berkesan serta kurang kesan sampingan adalah amat diperlukan. Sebatian fenolik telah dikenal pasti sebagai perencat XO yang berkesan dalam banyak kajian. Kajian in vitro dan in siliko telah dijalankan untuk mengkaji interaksi antara apigenin, kaempferol dan asid 4-hidrosibenzoik semasa perencatan XO. Apigenin didapati merupakan perencat XO yang paling berkesan dalam kalangan sebatian yang dikaji dengan skor dok yang terbaik sebanyak -8.2 kcal/mol sebagaimana yang ditunjukkan oleh simulasi dok molekul yang menunjukkan interaksi yang menggalakkan dengan enzim XO. Interaksi secara tambahan antara sebatian iaitu apigenin-kaempferol, apigenin-asid 4-hidroksibenzoik dan asid 4-hidroksibenzoik-kaempferol telah ditunjukkan dalam kajian in vitro dan in siliko. Hasil kajian menunjukkan asid 4-hidroksibenzoik-apigenin (-7.4 kcal/mol) adalah gabungan ligan yang paling stabil semasa didokkan pada XO. Simulasi dok berbilang ligan menunjukkan ligan bebas terikat pada tapak aktif XO di lokasi yang tidak mengganggu antara satu sama lain. Secara kesimpulannya, gabungan ketiga-tiga sebatian ini boleh diterokai dengan lebih lanjut dari segi interaksi tambahan mereka dalam perencatan XO, yang boleh dimanfaatkan dari segi peningkatan keberkesanan dan pengurangan kesan sampingannya dapat dipertingkatkan apabila setiap satu digunakan pada dos yang lebih rendah untuk memberikan kesan yang sama. 

 

Kata kunci: Dok molekul; interaksi tambahan; pelbagai ligan; perencat xantina oksidase; sebatian fenolik

 

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*Corresponding author; email: lohke@tarc.edu.my

 

 

 

 

 
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