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ANTIOXIDANT ACTIVITY OF ACTIVE COMPOUND FROM BANGLE RHIZOME (Zingiber purpureum) Ni Made P. Susanti1, I Putu D. N. Parahyangan1, Ida Ayu D.G. Swandari1 and Ni Putu S. Mahasuari1*

1Department of Pharmacy, Faculty of Mathematics and Natural Sciences, University of Udayana, Bukit Jimbaran, Badung, 80363, Indonesia E-mail : dekpitsusanti@unud.ac.id

Introduction Diabetes is a metabolic disorder disease caused by insufficient insulin produced by the pancreas or the body can not use insulin effectively so that blood glucose levels increase. Excess glucose is considered a major cause of tissue damage. Glucose can undergo auto oxidation that forms an active oxygen compound. The formation of such reactive oxygen compounds can improve the modification of lipids, DNA, and proteins in the tissues (Ueno et al., 2002). The molecular modification of this tissue produces an imbalance between protective antioxidants (antioxidant defenses) and increased free radical production. this is the beginning of oxidative damage known as oxidative stress (Nuttal et al., 1999). Oxidative stress due to free radicals has been shown to cause damage to cells that cause chronic diseases such as cancer, cardiovascular disease, inflammation and Alzheimer's disease (Halliwell and Gutteridge, 1989). Bangle Rhizome (Zingiber purpureum) has a terpenoid biomarker that is terpinen-4-ol which is known to have an antioxidant effect (Bhua-in and Paisooksantivatana, 2009; MOH, 2010). Antioxidant activity can be used to prevent oxidative stress in diabetic patients. Three other marker compounds in the bangle extract have been found in previous studies (Hartati et al., 2013). Based on this background, this research will be carried out molecular docking Zingiber purpureum marker with target protein SOD and GPX to prevent oxidative stress in diabetics through the mechanism as an antioxidant.

Abstract Tissue damage can occur in people with diabetes due to excess glucose that undergoes auto oxidation and forms active oxygen compounds. This causes an imbalance between protective antioxidants and increased free radical production known as oxidative stress. Bangle Rhizome (Zingiber purpureum) has terpenoid biomarker that is terpinen-4-ol which is known to have effect as antioxidant. Antioxidant activity can be used to prevent the occurrence of oxidative stress in patients with diabetes. Three other marker compounds in bangle extracts have been found in previous studies. This study aims to determine the activity of these compound to prevent oxidative stress in diabetics through the mechanism of antioxidant. Molecular docking performed includes the preparation of 3D structures of proteins using chimera 1.10.1 software, optimization of the 3D structure of test compounds using hyperchem 8 software (semi-empirical AM1), molecular docking method validation and molecular docking to the protein SOD (1MFM) and GPX (2F8A) using Autodock 4.2. software. The lower the binding energy, the stronger and more stable the bond between the protein and test compounds. The results showed that all test compounds are able to bind to proteins SOD and GPX with bond energy -4.83; -4.03; -4.68; -4.29; -4,39; -3.83; -3.57; -3.99 kcal/mol respectively. The interaction between the terpinen-4-ol and the three other compounds with the SOD and GPX proteins shows that these compounds have the potential to induce SOD and GPX proteins in capturing free radicals to prevent oxidative stress in patients with diabetes.

Keywords : Bangle Rhizome, Oxidative Stress, Antioxidant, In Silico

Methods Native ligand of the protein SOD (1MFM) and GPX (2F8A) obtained from http://www.rcsb.org/pdb/home/home.do. removed by Chimera 1.10.1 software to provide space (pocket/cavity), to determine the shape of the pocket, coordinate pocket, binding site center and radius cavity. Marker compounds of Zingiber purpureum structure are optimized by using Hyperchem 8 with semi-empirical AM1 method. Validation of molecular docking method carried out by redocking native ligands to proteins using Autodock 4.2 software. Docking method is valid if the value of RMSD (Root Mean Square Distances) ≤ 3 Å (Jain and Nicholls, 2008). The result of docking the compounds to SOD and GPX using Autodock 4.2. will show the conformation of compounds with the lowest binding energy to the target protein.

Result Preparation of protein SOD (1MFM) and GPX (2F8A) , downloaded from http://www.rcsb.org/pdb/home/home.do, aims to separate the native ligand and protein to provide space (pocket/cavity) during the docking process. The preparation is done using a chimera software 1.10.1. in order to obtain protein without native ligand (Figure 1). Optimization of the compound was carried out using a semiempirical AM1 in the Hyperchem 8 program and the total energy obtained by each compound are -2779.47 kcal / mol; -3565.32 kcal / mol; -3026,02 kcal / mol; and -4473.34 kcal / mol respectively (the structure can be seen in figure 2). This energy shows the amount of energy required to break one mole of bonding species in a gaseous state. The lower the total energy of the compounds, the more stable the three-dimensional structure of the compounds (Azam, 2012).

(A) (B) Figure 1. Protein structure (A) and the native ligand (B)

Terpinen-4-ol

Compound 1

Compound 2

Compound 3

(A)

(B)

Figure 3. Interaction between the compounds with protein SOD (A) and GPX (B)

Post docking result shows that the test compounds have antioxidant activity because they are able to bind to SOD and GPX with bond energy respectively -4,83 and -4,03 kcal/mol for terpinen-4-ol; -4,68 and -4,29 kcal/mol for Compound 1, -4,39 and -3,83 kcal/mol for Compound 2, and -3,57 and -3,99 kcal/mol for the Compound 3. The interaction between the terpinen-4-ol and the three other compounds with the SOD and GPX proteins shows that these compounds have the potential to induce SOD and GPX proteins in capturing free radicals to prevent oxidative stress in patients with diabetes.

Conclusion Marker compoundds from Zingiber purpureum have activity as antioxidant with inducing SOD and GPX proteins in capturing free radicals to prevent oxidative stress in patients with diabetes.

Acknowledgements This research was supported by Ministry of Research, Technology and Higher Education of the Republic of Indonesia. Poster Number : PP-NP-03

References Azam, F., A.M. Madi, and H.I. Ali. 2012. Journal of Young Pharmacists. Vol. 4 (3). Bhua-in, S. and Y. Paisooksantivatana. 2009. Kasetsart J. (Nat. Sci.). Vol. 43:467-475. Depkes RI. 2010 .Farmakope Herbal Indonesia. Jakarta : Departemen Kesehatan Republik

Indonesia. Halliwell, B., and J.M.C. Gutteridge. 1989. Free Radicals in Biology and Medicine, 2nd Etition.

Oxford : Oxford University Press. Hartati, S., Megawati, N. Artanti, L. Meilyawati, and M. Hanafi. 2013. Jurnal Ilmu Kefarmasiaan

Indonesia. Vol. 11(2):197-201. Jain, A.N. and A. Nicholls. 2008. J. Comput. Aided Mol. Des. Vol. 22:133-139. Nuttal, S.L., F. Dunne, M.J. Kendal, and U. Martin. 1999. Q J Med. Vol. 92:33-8. Ueno, Y., M. Kizaki, R. Nakagiri, T. Kamiya, H. Sumi, and T. Osawa. 2002. J Nutr. Vol.132:897-

900.

Validation aims to determine the validity of docking methods used RMSD (Root Mean Square Deviation) as parameter. RMSD values ≤ 3 Å stated that the experimental crystal structures resembling the native ligand crystallography in the PDB (Protein Data Bank) so that the molecular docking method is valid and accurate.. Docking the compounds on the protein SOD and GPX using Autodock 4.2 software, and coordinates was set according to the coordinates of the native ligand. The results of andrografolid docking with protein SOD and GPX shown in Figure 3.

Terpinen-4-ol Compound 1 Compound 2 Compound 3

(A)

(B)

Figure 2. 3-dimensional structure of the compounds before (A) and after (B) optimized

ANTIOXIDANT ACTIVITY OFACTIVE COMPOUND FROMBANGLE RHIZOME (Zingiber

purpureum)by Ni Made Pitri Susanti

Submission date: 16-Jul-2018 06:18PM (UTC+0700)Submission ID: 982872204File name: Poster-FIX.pptx (4.44M)Word count: 1473Character count: 6636

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