(A) DB00742 (B) DB03955
(C) DB04733
(D) DB03955
Fig 1: A,B,C,D shows ligand 2D structures
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In silico Interaction Analysis of Intracranial Pressure Reducing Agent Mannitol and its Derivatives with Human Serum Albumin
Usman Sayeed, Qazi Mohd. Sajid Jamal*, Mohd. Haris Siddiqui, Mohd. Salman Khan and Gulshan Wadhwa
Abstract— There is no specific treatment for Japanese encephalitis and treatment is supportive; with assistance given for feeding, breathing or seizure control as required. The recent data reported that mannitol (MNT) diluted into the human serum reduce the intracranial pressure, which may be effec- tive in intracranial pressure management. Therefore, the present study was designed to find out the molecular interaction of mannitol with human se- rum albumin (HSA). Docking results showed that mannitol was efficiently bounded with HSA. HSA Pdb Id: 1E7H docked with Mannitol like Com- pounds DB00742, DB03955, DB04733, and DB03206 was -6.65 Kcal/Mol. -5.46 Kcal/Mol.and -4.54 Kcal/Mol and -6.02 Kcal/Mol, respectively. Study re- veals that in silico approach can easily explore the molecular interaction of mannitol with HSA and it will lead to understand the binding pattern of mannitol with HSA.
Index Terms— Mannitol (MNT), human serum albumin (HSA), Japanese encephalitis, molecular interaction.
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annitol is used clinically in osmotherapy to condense strongly raised intracranial pressure until more defini-
tive treatment can be applied, e.g., after head trauma. It is also
used to treat patients witholiguric renal failure. It is adminis- tered intravenously, and is filtered by the glomeruli of the kidney, but is incapable of being resorted from the renal tubule, resulting in decreased water and Na+reabsorption via its osmotic effect. Consequently, mannitol increases water and Na+ excretion, thereby decreasing extracellular fluid volume [1].
Mannitol can also be used as a facilitating agent for the trans- portation of pharmaceuticals directly into the brain. The arter- ies of the blood–brain barrier are much more selective than normal arteries. Normally, molecules can diffuse into the tis- sues through gaps between the endothelial cells of the blood vessels. However, what enters the brain must be much more rigorously controlled. The endothelial cells of the blood–brain barrier are connected by tight junctions, and simple diffusion through them is impossible. Rather, active transport is neces- sary, requiring energy, and only transporting molecules that the arterial endothelial cells have receptor signals for. Manni- tol is capable of opening this barrier by temporarily shrinking the endothelial cells, simultaneously stretching the tight junc- tions between them [2]. An intracarotid injection of high mo- larity mannitol (1.4-1.6M), causes the contents of the artery to
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1Department of Biosciences and Bioengineering, Integral University, IIT Dasauli, Kursi
Road, Lucknow-226026, Uttar Pradesh, India. E-mail: usman.sayeed@gmail.com
3Department of Biotechnology, Ministry of Science and Technology, Government of India, CGO Complex, Lodhi Road, New Delhi- 110003,India.
be hyperosmotic to the cell.
Water leaves the cell and enters the artery in order to recreate an osmotic equilibrium. This loss of water causes the cells to shrivel and shrink, stretching the tight junctions between the cells [3]. The newly formed gap reaches its peak width five minutes after mannitol injection, and stays widely open for thirty minutes. During this time span, drugs injected into the artery can easily diffuse through the gaps between cells direct- ly into the brain [4]. This makes mannitol indispensable for delivering various drugs directly to the brain (e.g., in the treatment of Alzheimer's disease, or in chemotherapy for brain tumors [3].
Current study explores the binding site of mannitol and its derivative on human serum albumin through molecular dock- ing studies.
Ligand files of Mannitol (DB00742), 1,5-Dideoxy-1,5-Imino- D-Mannitol (DB03955), 1,6-Di-O-Phosphono-D-Mannitol (DB04733), 1-Deoxynojirimycin (DB03206) were downloaded in .mol format (Fig:1) from DrugBak Database (http://www.drugbank.ca) [5]. These files could not directly use by Autodock 4.2 tools [6] thus; we have to convert it into
.pdb files and also further the ligands were submitted for
CHARMm [7] energy minimization protocol in Discovery
Studio 4.1.
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(A) DB00742 (B) DB03955
(C) DB04733
(D) DB03955
Fig 1: A,B,C,D shows ligand 2D structures
Human serum albumin (HSA) PDB ID: 1E7H [8] was obtained from Protein Data Bank (Fig: 2). Published structures were edited to remove HETATM ,further the protein structure was submitted for CHARMm [7] energy minimization protocol using Discovery Studio 4.1.
Fig 2: Crystal 3D structure of Human Serum Albumin (PDB ID: 1E7H)
Windows 7 professional Version 2002, Service pack 3 operat- ing System on Intel (R) Core (i3) , CPU T6500 @ 2.10 GHz, 1.19
GHz, and 4.00 GB of RAM of Acer Machine. Molecular dock- ing methods followed by searching the best conformation of HSA and Mannitol like Compounds DB00742, DB03206, DB03955 and DB04733 on the basis of binding energy. Water molecules were removed from the protein structures before docking and hydrogen atoms were added to all target pro- teins. Kollman united charges and salvation parameters were added to the proteins. Gasteiger charge was added to the lig- ands. Grid box was set to cover the maximum part of proteins
and ligand. The values were set to 60×60×60 Å in X, Y and Z
axis of a grid point. The default grid points, spacing was 0.375
Å. Lamarckian Genetic Algorithm (LGA) [10] was used for
proteins ligands flexible docking calculations. The LGA pa- rameters like population size (ga_pop_size) , energy evalua- tions (ga_num_generation) , mutation rate , crossover rate and step size were set to 150, 2500000 , 27000 , 0.02 , 0.8 and 0.2 Å
,respectively. The LGA runs were set at 40 runs. All obtained
40 conformations of proteins and ligand complex were ana- lysed the interactions and binding energy of the docked struc- ture using Discovery Studio Visualizer (version 4.1).
We have taken Human serum albumin (HSA) Pdb Id: IE7H as a receptor (Fig: 2) for the docking analysis. Furthermore, we have observed that the binding energy between HSA and Mannitol like Compounds DB00742, DB03955, DB04733, and DB03206 was -6.65 Kcal/Mol. -5.46 Kcal/Mol.and -4.54
Kcal/Mol and -6.02 Kcal/Mol, respectively (Table :1).
The observed inhibition constant (Ki) for DB00742, DB03955, DB04733, and DB03206 was 13.28 uM, 100.14 uM, 471.56 uM and 13.28 uM, respectively (Table : 1).
We have also found that the 10 hydrogen bonds between and
DB00742 HSA (Fig :3) , 5 hydrogen bonds between DB03955 and HSA (Fig: 4), 7 hydrogen bonds between DB04733 and HSA (Fig: 5) and 10 hydrogen bonds between DB03206 and HSA (Fig: 6).
Docking studies were performed by Autodock (Version 4.2)
suite [6],[9] and Cygwin interface was used in the Microsoft
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Fig. 3 : shows Amino acid residues involved in hydrophobic in- teraction in DB00742 and HSA interaction.
Fig. 6 : shows Amino acid residues involved in hydrophobic in- teraction in DB03206 and HSA interaction
Fig. 4 : shows Amino acid residues involved in hydrophobic in- teraction in DB03955 and HSA interaction
Fig. 5 : shows Amino acid residues involved in hydrophobic in- teraction in DB04733 and HSA interaction
TABLE 1
OBSERVED DOCKING ANALYSIS RESULTS
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As we know that every drug development study de- pend on the Clinical Trial Designs of Therapeutic Inter- ventions. All techniques are time taking and need a lot of financial support. With the help of in silico approach we can reduce the time and cost of the clinical trials and make our study specific and concise on specific targets. Struc- ture based techniques like docking analysis play im- portant role to understand molecular dynamic properties of the ligands and receptor interactions and we can easily analyze the properties of binding pockets.
Furthermore, our study suggests that mannitol and its derivatives could be used as a primary treatment in the case of Japanese encephalitis like diseases related to blood brain barriers. In vivo and In vitro validation is needed to authenticate in silico results reported in the current study.
The authors are thankful to the Prof. S.W. Akhtar , Hon’ble Vice-Chancellor,Integral University for providing necessary infrastructure facility to carry out the study.
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