A DISSERTATION ON

Hypocholesterolemic Potential of S-(2-carboxypropyl)-glutathione and S-allyl-

marcapto glutathione from Allium sativum (Linn.): An in-silico study

Presented by Supervisor
INAMUL HASAN Dr. M. SALMAN KHAN
B. Sc. Life Sciences, 6 th Sem. Associate Professor
Department of Biosciences Department of Biosciences
Integral University, Lucknow. Integral University, Lucknow
 INTRODUCTION

 Cardiovascular disease (CVD) is the major cause of death in

developed and developing countries.

 As estimated by the World Health Organization (WHO) in 2016,

17.9 million people died from CVDs in 2012, representing 31%
of all global deaths.

 CVD include many diseases related to heart and blood vessels,

e.g., coronary heart disease, atherosclerosis and hypertension.

 Hyperlipidemia is a condition of excess fatty substances called

lipids, largely cholesterol (TC) and triglycerides (TG), in the
blood.
(A) Ultra structure of arteries (B) Cartoon representing how cholesterol
accumulation interrupts the normal blood
flow.

Figure 1
 There are number of risk factors which induce the
atherosclerosis
Such as
 Hypercholesterolemia,
 Cholesterol induced oxidative stress,
 LDL-cholesterol,
 Infection and inflammation,
 Hypertension,
 Tobacco use,
 Cigarette smoking and diabetes.

Several studies have identified LDL as independent risk factors that modulate
CVD risks.
High levels of plasma LDL-C are directly related to the development of CVD,
whereas, an inverse association between the incidence of CVD and high
concentrations of HDL-C has been observed.
 HMG-R
• HMG-R is the rate limiting enzyme of the cholesterol biosynthetic pathway.

• Human HMG-R consists of a single polypeptide chain of 888 amino acids. The N-
terminal (339 residues) is membrane bound and resides in the endoplasmic reticulum
membrane.

• While the catalytic activity of the protein resides in its cytoplasmic soluble C-terminal
portion (residues 460-888).

Figure 2 : HMG-R contains four identical chains. Chain Figure 3 : Representation of Chain A of
A –green; chain B – purple, chain C - gray, chain D- HMG-R.
Light Blue.
 Therapeutic approaches against atherosclerosis

 Dietary changes
 Weight loss
 Regular exercise
 Quitting smoking
 Medications such as statins and
 Periodic lipid screenings

Statins (or HMG-CoA reductase inhibitors) are a class of drugs used to decrease
cholesterol levels by inhibiting the activity of HMG-CoA reductase, which plays a
central role in the production of cholesterol.

Increased cholesterol levels have been associated with cardiovascular diseases

(CVD), and statins are therefore used in the prevention of these diseases.

Limitations of statins

Statins are synthetic drugs, long term use of statins have host of side effects and
incurve problems- such as
• Muscle damage Statin intolerance
• Toxicity
• Cost
GARLIC AND OSCs
 Garlic (Allium sativum Linn), a member of Allium genus.

 It has numerous beneficial biological activities i.e. lipid-lowering, anti-

inflammatory, antidiabetic, anticancer and anti-atherosclerotic properties.

 These properties are attributed to its rich content of different volatile organosulfur
compound (OSCs) and other phytochemicals vis. phenolic acids, flavonoids,
thiosulfonates etc.

 Among all the dietary phytochemicals from A. sativum Linn, OSCs have been
found to show greater effects against various metabolic disorders.

 Major OSCs from garlic with high therapeutic and pharmacological properties are:

 Allicin,
 DAD,
 DAT,
 Allyl methyl trisulfide,
 Ajoene and many others.
 OBJECTIVES

I. To perform in-silico molecular interaction study of S-(2-carboxypropyl)-

glutathione against active pocket of β-hydroxyl- β-methyl glutaryl CoA
reductase.

II. To perform in-silico molecular interaction study of S-allyl-marcapto

glutathione against active pocket of β-hydroxyl- β-methyl glutaryl CoA
reductase.

III. To perform in-silico molecular interaction study of S-Propylcysteine against

active pocket of β-hydroxyl- β-methyl glutaryl CoA reductase.

IV. To perform in-silico molecular interaction study of N-Acetyl-S-allylcysteine

against active pocket of β-hydroxyl- β-methyl glutaryl CoA reductase.

V. To perform in-silico molecular interaction study of standard drug

Atorvastatin against active pocket of β-hydroxyl- β-methyl glutaryl CoA
reductase.
Results
Figure 3: Molecular interaction of S-(2-
carboxypropyl)-glutathione (CID: 22837928)
with the active pocket of β-hydroxyl- β-
methyl glutaryl CoA reductase (1DQ9).

Figure 4: Panel (a)- Formation of hydrogen

bonds to stabilize the S-(2-carboxypropyl)-
glutathione-HMG-R complex; Panel (b)-
Hydrophobic surface around S-(2-
carboxypropyl)-glutathione-HMG-R
complex.
Figure 5: Molecular interaction of
S-allyl-marcapto glutathione
(CID: 9907834) with the active
pocket of β-hydroxyl- β-methyl
glutaryl CoA reductase (1DQ9).

Figure 6: Panel (a)- Formation of

hydrogen bonds to stabilize the S-
allyl-marcapto glutathione-HMG-
R complex; Panel (b)-
Hydrophobic surface around S-
allyl-marcapto glutathione-HMG-
R complex.
Figure 7: Molecular interaction
of S-Propylcysteine (CID:
125198) with the active pocket
of β-hydroxyl- β-methyl
glutaryl CoA reductase (1DQ9).

Figure 8: Panel (a)- Formation

of hydrogen bonds to stabilize
the S-Propylcysteine-HMG-R
complex; Panel (b)-
Hydrophobic surface around
S-Propylcysteine-HMG-R
complex.
Figure 9: Molecular interaction of
N-Acetyl-S-allylcysteine (CID:
152467) with the active pocket of β-
hydroxyl- β-methyl glutaryl CoA
reductase (1DQ9).

Figure 10: Panel (a)- Formation of

hydrogen bonds to stabilize the N-
Acetyl-S-allylcysteine-HMG-R
complex; Panel (b)- Hydrophobic
surface around N-Acetyl-S-
allylcysteine-HMG-R complex.
Figure 11: Molecular
interaction of standard drug
Atorvastatin (CID: 60823) with
the active pocket of β-hydroxyl-
β-methyl glutaryl CoA
reductase (1DQ9).

Figure 12: Panel (a)- Formation

of hydrogen bonds to stabilize
the Atorvastatin-HMG-R
complex; Panel (b)-
Hydrophobic surface around
Atorvastatin-HMG-R complex.
Table 1: Molecular interaction studies of various organosulfur compounds
against the active pocket of HMG-R.

Compound Name Compound Binding energy Residues involved

ID (ΔG)
(Kcal/mol)
S-(2-carboxypropyl)- 22837928 -5.20 Arg590, Ser684, Cys688, Asn686,
glutathione Asp690, Lys691, Lys692, Ala769,
Pro693, Val772
S-allyl-marcapto 9907834 -4.74 Lys662, Ala525, Ala658, Met659,
glutathione Met657, Gly656, Ala654, Met655,
Gly803, Gly765, Gln766, Val805,
Gly807, Gly806, Gly808, Thr809,
Asn810
S-Propylcysteine 125198 -4.41 Ala654, Met655, Gly656, Met657,
Asn658, Met659, Gly765, Gln766,
Asp767, Gly803, Val805, Gly806,
Gly807, Thr809
N-Acetyl-S- 152467 -3.79 Met655, Gly656, Met657, Asn658,
allylcysteine Gln766, Asp767, Ile802, Gly803,
Gly806, Gly807
Atorvastatin 60823 -4.86 Ile536, Met534, Thr758, Val772,
Asn771, Ala768, Asp767, Gly765,
Gln814, Gly808, Leu811, Ile762
 CONCLUSION

 Our results from the present study demonstrated that among all OSCs used in this
study, S-(2-carboxypropyl)-glutathione and S-allyl-marcapto glutathione) showed the
best binding energy while occupying the active pocket of HMG-R (-5.20 Kcal/mol
and – 4.74 Kcal/mol, respectively), whereas, rest of the OSCs also showed their
interaction with the similar residues of active pocket of HMG-R.

 Based on above findings, we concluded that garlic based diet may be promoted to
fight with cholesterol induced oxidative stress and hypercholesterolemia in the
patients suffering with inadequate lipid lowering with statins.

 These OSCs may also be beneficial to the patients suffering with long term statin
therapy and subsequent adverse effects.