INTRODUCTION:

Diabetes mellitus is among the most common and chronic lifestyle diseases in the world and the number of diabetic patients is on the rise^1,2. It is a metabolic disorder in which the body does not produce or properly use insulin^3,4. The World Health Organization (WHO) reports that about 200 million people worldwide are living with diabetes, which is expected to double by 2030. WHO says that 80 per cent of people die every year due to diabetes in middle-income countries⁵. The newly published Indian Council for Medical Research-India Diabetes (ICMR-INDIAB) national study estimated that there have been 62.4 million people with type 2 diabetes (T2DM) and 77 million people with pre-diabetes in India⁶. This would be increased to 100 million by 2030⁷. T2DM primarily affects older people in developed countries, while in developing countries such as India it affects younger people at the height of their working lives and thus presents even larger risks to health of these individuals⁸.

Several Indian traditional medicines have been investigated for their beneficial use in diabetes and studies on the merits of their use are published in various scientific journals ⁹.

Andrographis paniculata (Acanthaceae) is one of the most valuable medicinal plants and bio-factories of diterpronidlactons of tremendous value, such as immune-stimulating, anti-inflammatory, anti-fertility, liver protection, anti-HIV and bile-stimulating agents^10,11. A number of active components are identified in this plant, which mainly includes diterpronidlactons, flavonoids and polyphenols^12,13.

Glucokinase (GK) is a cytoplasmic monomeric enzyme present in the liver and pancreas¹⁴. Its main function is to regulate the amount of glucose in these organs¹⁵. In hepatocytes, glucose phosphorylation by glucokinase promotes glucose absorption and metabolism by retaining a gradient for glucose transport to these cells, thereby regulating the elimination of hepatic glucose¹⁶. GK is thought to be part of the glucose-sensing system in beta cells and to be involved in the regulation of insulin release. During diabetes, complete or partial insulin deficiency causes carbohydrate metabolism disorders that reduce the activity of several main enzymes such as glucokinase, resulting in impaired glucose utilisation and increased hepatic glucose output¹⁷. Hence the enzyme GK could be the drug target for type 2 diabetes (T2D) and its activation could result in lower blood glucose levels regardless of the cause of hyperglycemia¹⁸. The goal of this study was therefore to identify the effective compounds that activates Glucokinase from Andrographis paniculata compounds through a molecular docking analysis.

MATERIALS AND METHODS:

Protein Preparation:

The structures of the target receptor of human glucokinase (PDB: 1V4S), was downloaded from protein databank. Before using this structure for docking studies, the heteratoms, water molecules and co-crystallized ligands were removed and then it was saved as pdbqt format.

Ligand Preparation:

12 compounds from Andrographis paniculata were retrieved from PubChem compound database. These molecules were downloaded in Structure Date File (SDF) format and converted to Protein Data Bank (PDB) coordinates by using online smiles translator.

Molecular Docking:

PyRx Virtual Screening Technology was used in the virtual screening of ligands like AutoDock ¹⁹ and AutoDock Vina²⁰ with the Lamarckian Genetic Algorithm (LGA) as a score function. Active site dimensions have been configured as a grid for the dock of ligands at which a maximum of 10 has been determined for each ligand. Prior to the start of the docking process, charges were assigned to the protein and ligand structures of AutoDock Vina. The resulting ligands have such a significant potential to be used as drug candidates.

RESULTS AND DISCUSSION:

Many herbs and plant products have been shown to have antihyperglycemic action ^21,22. The valuable traditional medicinal plant Andrographis paniculata (Nees) has numerous significant bioactive components. In humans, it treats and prevents a variety of illnesses ^23,24. Hence the present study was designed to study the molecular interactions of selected compounds with diabetes mellitus (DM) target protein Glucokinase. Docking simulations at 1V4S active sites have been performed by the Auto Dock-PyRx software, that have been shown to successfully replicate experimentally measured binding modes in terms of the lowest docking energy. The target protein structure of 1V4S was docked with Andrographis paniculata compounds, which produced good results as shown by the lowest binding energy values. The highest suitable binding modes of all ligands at the target protein active sites have been shown using PYMOL. Hydrogen-binding interaction of selected compounds to target protein and their corresponding energy values are shown in Table 2.

Table 1: List of selected compounds from Andrographis paniculata

S. No	Compound Name
1	3,14,19-triacetylandrographolide2D_CID_25121277
2	14-acetylandrographolide2D_CID_71589914
3	14-deoxy-11,12-didehydroandrographolide_CID_5708351
4	14-deoxy-14,15-didehydroandrographolide_CID_6473762
5	14-deoxyandrographolide_CID_11624161
6	Andrograpanin_CID_11666871
7	Andrographolide_CID_5318517
8	Isoandrographolide_CID_49841562
9	Neoandrographolide_CID_9848024
10	phytol_CID_5280435

Figure 1a demonstrates the outcome of human glucokinase (1V4S) docking analysis with 3, 14,19-triacetylandrographolide. Glucokinase amino acid residues such as THR-82, ARG-85, LYS-169 THR-228 and SER-445 were formed by H-bond with 3,14,19-triacetylandrographolide. It also demonstrates strong binding energy—6.8kcal / mol. Figure 1b showed that Andrographolide exhibited significant interactions with glucokinase protein. It was formed by four H-bonds, i.e. THR-82, SER-161, ASP-305 and SER-411. Andrographolide also exhibited a strong bond with glucokinase in terms of binding energy. It Showed a binding energy of-7.2 kcl / mol.

Figure 1c shows that 14-deoxyandrographolide also exhibited heavy interaction with 1V4S through the formation of H-bonds with ASP-78, ARG-85, LYS-169, SER-1517THR-228 residues. The binding energy of the complex was-7.3 kcal / mol. Figure 1d shows that Andrograpanine formed H-bonds with THR-228, GLU-443, GLY-444 and SER-445. It indicates that the binding energy was-6.9 kal / mol. Figure 1E depicted that Neoandrographolide exhibited very strong interactions with glucokinase through H-bond interactions with LYS-169, THR-228, GLY-229, ASP-409, LYS-414 and GLU-443. It formed six hydrogen bond interactions and also showed good binding energy i.e -7.2.

Figure 1: Molecualr interaction of Glucokinase with a) 3,14,19-triacetylandrographolide b) Andrographolide c) 14-deoxyandrographolide d) Andrograpanin e) Neoandrographolide

Molecular docking studies are widely accepted for drug screening ^25-27 Here, the compounds identified from the Andrographis paniculata in the current study showed the strong activity against glucokinase protein in terms of binding energy and hydrogen bond interaction. Hence, it is evident from the analysis that the compounds from Andrographis paniculata can also be used as an anti- diabetic agent after experimental validation.

CONCLUSION:

Docking studies of the ligand Andrographis paniculata with glucokinase target protein showed that compounds such as 3,14,19-triacetylandrographolide, Andrographolide, 14-deoxyandrographolide, Andrograpanin and Neoandrographolide dock well with target related to diabetes mellitus. Hence, Andrographis paniculata may play an important role in blood glucose regulation via activation of GK.

CONFLICT OF INTEREST:

The authors have no conflicts of interest.

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Received on 21.03.2024 Revised on 19.06.2024

Accepted on 02.08.2024 Published on 24.12.2024

Available online from December 27, 2024

Research J. Pharmacy and Technology. 2024;17(12):5981-5983.

DOI: 10.52711/0974-360X.2024.00907