Investigation Of Plant Based Angiotensin

Converting Enzyme Inhibitor For Hypertension

University School of Biotechnology

Guru Gobind Singh Indraprastha University
Dwarka Sector – 16C, Delhi - 110078

Presented by – Guide
Simranjeet Singh Mann Dr. K. K. Aggarwal
03716001316 Professor
 Hypertension

 When the blood pressure increases and become more than 140/90
mmHg, then a person is said to have hypertension (Tabassum et al.,
2010)

Source: https://www.sprucemedicalgroup.com

Primary Hypertension Secondary Hypertension

 Cause:-
 potassium deficiency,
alcohol intake, aging
inherited genetic mutations,
increased amount of
angiotensin 2
 Cause :-
 compromised/imbalanced
pathophysiological mechanism
like hormone-regulating
endocrine system and heart
function
 Impact on society

 In 2000, almost 1 billion of the world adult population had HT and more than 20
million had heart failure (Whelton et al., 2005)
 Hypertension has become the main cause for premature mortality and
cardiovascular illness with more prevalence in LMICs than HIC (Mills et al.,
2020)

People affected with Hypertension in billion %death due to Hypertension

1.2
1.04
1
19 24
No. in Billions

0.8
0.6
0.4 0.35 57

0.2
0
HICs LMICs Coronary Heart Disease death Stroke Deaths Other

Data obtained from Mills et al., 2020 and Gupta et al., 2004
 Causes and number of worldwide death due to high blood pressure

6000
Attributed to SBP>110-115
mmHg
5000 4862
Attributed to SBP>140
mmHg
Deaths in thousands

4000
3573

3000

1953
2000
1489 1423 1552
1134 1151
1000 848
553

0
Ischaemic Isachaemic Haemorrhagic Other Chronic
Heart Disease Stroke Stroke cardiovascular Kidney disease
disease

Types of Diseases
Data obtained from Mills et al., 2020
 Mechanism of Hypertension
 RAAS (Renin-Angiotensin –
Aldosterone System)

 Elements are Renin, ACE,

Angiotensinogen,
Angiotensin1, Angiotensin 2,
Aldosterone, AT1 and AT2
Receptor

 Regulates body hemodynamic

equilibrium ,circulating volume
and electrolyte balance. (Lappin et
al., 2021)
 Angiotensin Converting Enzyme(ACE)
PDB ID:1O86
 ACE (EC3.4.15.1) is a two domain
dipeptidyl carboxy peptidase involved in
regulation of blood pressure.

 Active sites: Glu 348

 Expression is in endothelium and

epithelial cell surfaces

 Hydrolyzes angiotensin 1
angiotensin 2 (vasoconstrictor) by release
of the C-terminal His-Leu

Fig. Structure Of Human Somatic Angiotensin 1

Converting Enzyme (source: RCSB PDB)
Angiotensin 2

 Vasoconstrictor
 Stimulates adrenal gland
cortex
 Lipogenesis
 Increased Calcium influx

Source: RCSB PDB

 Current ways to fight Hypertension

• Increased potassium intake

Non • Reduced salt intake
Pharmacological • Controlled alcohol consumption
• Regular exercise
ways • Weight loss

• ACE inhibitors like enalapril, Ramipril , etc.

• Angiotensin 2 receptor blockers like
Pharmacological losartan,telmisartan, etc.
• Dihydropyridine calcium channel blockers
ways • Thiazide-type and thiazide-like diuretics

Source:
Source: (Aburto
(Aburto et
et al.,
al., 2013)(Appel
2013)(Appel L
L et
et al.,
al., 2009)(Garjon
2009)(Garjon et
et al.,
al., 2013)
2013)
Side effects of current medication

Cough

Elevated blood potassium levels

Low blood pressure,

dizziness
Headache

Drowsiness

Weakness

Abnormal taste (metallic or salty taste)

Rash

Chest pain

Increased uric acid levels

(Source: Oparil et al., 2018)

 Rationale

 Current treatments for hypertension have some adverse effects.

 Plant based remedy can be effective option for the treatment of hypertension.

 Natural medicines from plants have better acceptance to human body with less
side effects and are also reported to have antihypertensive effects

 Around 80% of the world population use natural medicines as they have better
acceptance by the human body with less side effects. (Oparil et al., 2018)

 ACE is one of the most important enzyme of RAAS, thus, by inhibiting its
action, hypertension can be controlled.
 Objectives

1. Search for Angiotensin Converting Enzyme inhibitors

from plants.

2. To assess the potential of screened inhibitor as drug

therapeutic by in silico-studies.
 Methodology
Screened for various metalloprotease inhibitors of ACE from existing literature
1 reviews.

2 Performed protein blast using pBLAST to find the inhibitors in plants but no
similar proteins were found

Screened for various plants and its chemical compounds with

3 antihypertensive nature from existing literature.

Retrieved the “.sdf" files of the particular inhibitors that are found in
4 plants from PubChem.

5 Used OpenBabel software to convert the “.sdf” file to “.pdb” files

Using the AutoDock 4.2 performed docking of the inhibitors with ACE and
6 existing inhibitors as a control.

7 Used the PLIP server to analyze the interactions between inhibitor and ACE.
Result
 Proteinaceous inhibitors against ACE

S.NO. Name Source PDB ID

1 TIMP1 Homo sapiens 1D2B

Homo sapiens
2 TIMP2 1BR9

Homo sapiens
3 TIMP3 3CKI

Homo sapiens
4 TIMP4 -
pBlast for proteinaceous inhibitors
Potential chemical inhibitor against ACE

S.No. Name Plant Source PubChem ID

Clerodendrum
1 Verbascoside 354009
bungei

Clerodendrum
2 Isoacetoside 6476333
bungei

Luteolin
3 Apium graveolens 5280445
 Apium
graveolens

Clerodendrum
bungei

Source: www.google.com
Interaction between ACE and chemical
compounds
ACE residues interaction with Isoacetoside
ACE residues interaction with Luteolin
ACE residues interaction with Verbascoside
ACE residues interaction with Enalapril(control)
 Docking results
Inhibition
Binding Energy
Potential inhibitors constant Amino Acid involved in interaction
(Kcal/Mol)
(Ki)

Enalapril HIS383(A), HIS387(A), PHE457(A),

-8.14 1.08uM
(control) TYR523(A), PHE527(A)

ALA149(A), GLN160(A), LEU161(A),

Luteolin -10.51 19.89 nM TRP185(A), LYS343(A), GLU349(A),
ARG348(A), VAL350(A)

SER284(A), TYR287(A), ASP288(A),

Isoacetoside -11.18 6.35 nM VAL291(A), SER298(A), LEU375(A),
GLU376(A), LYS449(A)

LYS118(A), SER219(A), SER222(A),

MET223(A), GLU403(A), GLY404(A),
Verbascoside -16.76 519.68 fM
ALA405(A), PRO407(A), HIS410(A),
GLU411(A), ARG522(A)
 Conclusion
 The compound docked with ACE showed binding and are able to inhibit
the activity of ACE and can be used to control hypertension
 The binding energy of control and screened inhibitors were compared.
The selected screened inhibitors have shown good result with respect to
control.
 The order of binding energies(Kcal/mol) of the inhibitors with ACE is as
follows:-
-Vebascoside(-16.76)> Isoacetoside(-11.18)>Luteolin(-10.51)>
Enalapril(control)(-8.14)
 Therefore these compounds can be taken into consideration for further
experiments as drug to tackle hypertension
 Future Aspects

 Further experiments of these compound are need to be

performed in the laboratory to test their correct efficiency
as an inhibitor against ACE.

 Further validation of docked complexes may be done by

performing MD simulation studies.
Acknowledgement