MOLECULAR DOCKING AND VIRTUAL SCREENINF OF ANTI-DIABETIC DRUG

1. INTRODUCTION
1.1 Drug Discovery and Development: -

Drug discovery and development is a scientific process through which new medicines
are identified, evaluated, and brought to the market for therapeutic use. It is a highly
complex and multidisciplinary field involving chemistry, biology, pharmacology, and
medical sciences. The aim is to develop drugs that are safe, effective, and of high
quality for the treatment and prevention of diseases such as Cancer and infectious
disorders.

Drug discovery and development is a systematic and lengthy process used to

identify, test, and bring new medicines to market. It begins with the identification of a
biological target such as a protein, gene, or receptor that is associated with a disease.
Once a target is identified, scientists validate it to confirm that modifying it will have
a therapeutic effect. After validation, the process moves to the discovery phase where
thousands of chemical compounds are screened to find “hit” molecules that show
potential activity against the target.

These hit compounds are then optimized to improve their effectiveness, safety,
and drug-like properties, leading to the selection of a promising “lead” compound.
The next stage is preclinical testing, where the lead compound is tested in laboratory
experiments and animal models to evaluate its toxicity, pharmacokinetics, and overall
safety profile. If the results are satisfactory, the drug enters clinical trials, which are
conducted in humans.

Clinical trials are carried out in three main phases. Phase I trials involve a
small number of healthy volunteers to assess safety and dosage. Phase II trials are
conducted on a larger group of patients to evaluate the drug’s effectiveness and side
effects. Phase III trials include a much larger population to confirm effectiveness,
monitor adverse reactions, and compare the new drug with existing treatments. If the
clinical trials are successful, the data is submitted to regulatory authorities for
approval.

Regulatory agencies such as the FDA or CDSCO carefully review all

preclinical and clinical data to ensure the drug is safe and effective for public use.
Once approved, the drug is manufactured on a large scale and marketed for medical

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use. Even after approval, the drug continues to be monitored in the post-marketing or
Phase IV stage to detect any long-term or rare side effects in the general population.

1.2 Drug Development Stages: -

Fig no.1 Drug Development Stages

Drug development is a long and complex process used to discover, test, and bring a
new medicine to market. It ensures that drugs are safe, effective, and of good quality
before being used by patients.

1. Drug Discovery & Development

This is the initial stage where scientists identify a disease target (such as a protein or
gene) and search for potential drug molecules.

• Target identification (finding disease-related molecules)

• Target validation (confirming its role in disease)
• Screening of thousands of compounds
• Selection of promising “hit” compounds

2. Preclinical Research

Before testing in humans, the drug is tested in the laboratory and on animals.

• Laboratory (in vitro) testing

• Animal (in vivo) testing

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• Study of toxicity, safety, and dosage

• Evaluation of pharmacokinetics (absorption, distribution, metabolism,
excretion)

3. Clinical Trials (Human Testing)

This stage tests the drug in humans and is divided into 3 phases:

• Phase I
Small group (20–100 healthy volunteers)
Focus: Safety and dosage

• Phase II
Larger group (100–300 patients)
Focus: Effectiveness and side effects

• Phase III
Large population (1,000–3,000+ patients)

Focus: Confirmation of effectiveness, monitoring adverse reactions

4. Regulatory Approval

• Government agencies review all data to ensure safety and effectiveness.

• Agencies: FDA (USA), CDSCO (India), EMA (Europe)
• Review of clinical and preclinical data
• Approval is granted if standards are met

5. Manufacturing & Marketing

Once approved, the drug is produced on a large scale and made available to the
public.

• Large-scale production
• Quality control
• Distribution to pharmacies and hospitals
• Marketing and awareness

6. Post-Market Surveillance (Phase IV)

Even after approval, the drug is continuously monitored.

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• Detect rare or long-term side effects

• Monitor safety in large populations
• Improve usage guidelines if needed

1.3 CADD (COMPUTER AIDED DRUG DESING) :-

Computer-Aided Drug Design (CADD) is a computational approach used in modern

pharmaceutical research to discover, design, and optimize new drug molecules with
the help of computer software and simulation techniques. It plays an important role in
reducing the time, cost, and experimental effort required in traditional drug discovery
processes. CADD integrates knowledge from chemistry, biology, pharmacology, and
computer science to predict how small molecules interact with biological targets such
as proteins, enzymes, or receptors.

CADD is broadly classified into two main types: structure-based drug design
(SBDD) and ligand-based drug design (LBDD). Structure-based drug design is used
when the three-dimensional structure of the target protein is known. In this method,
researchers study the active site of the protein and design or screen molecules that can
fit into the binding pocket effectively. Techniques such as molecular docking and
molecular dynamics simulation are commonly used to analysed the interaction
between the drug and the target protein. Ligand-based drug design is used when the
structure of the target protein is not available but information about known active
compounds exists. In this approach, scientists analysed existing molecules that show
biological activity and identify common chemical features responsible for their
effectiveness. Methods such as pharmacophore modelling and quantitative structure–
activity relationship (QSAR) are used to predict the activity of new compounds based
on structural similarity.

The CADD process typically begins with target identification, where a

disease-related biological molecule is selected. This is followed by target structure
preparation, in which the three-dimensional structure of the protein is obtained from
experimental databases or computational modelling. After this, large chemical
libraries containing millions of compounds are prepared for virtual screening. Virtual
screening is a computational technique used to filter and identify the most promising
drug-like molecules from large databases.

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The selected compounds are then subjected to molecular docking studies to

predict their binding orientation and affinity with the target protein. The best-
performing compounds are further optimized to improve their potency, selectivity, and
safety. In addition, ADMET prediction is performed to evaluate absorption,
distribution, metabolism, excretion, and toxicity properties of the drug candidates
before experimental testing.

CADD is widely used in modern drug discovery for diseases such as cancer,
viral infections, bacterial infections, and neurological disorders. It has significantly
contributed to the development of antiviral drugs, including drugs used in HIV and
COVID-19 treatment research. Pharmaceutical industries extensively use CADD tools
such as AutoDock, Schrödinger Suite, MOE, and Discovery Studio for drug screening
and optimization. One of the major advantages of CADD is that it reduces the need
for extensive laboratory experiments by predicting the most promising compounds in
advance.

Fig no.2 Computer Aided Drug Design

1.4 Molecular Docking: -

Molecular docking is a computational technique used in Computer-Aided Drug

Design (CADD) to predict the preferred orientation of a small molecule (ligand) when
it binds to a target protein or receptor. It helps scientists understand how a drug
molecule fits into the active site of a protein and how strong the interaction is. The

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main goal of molecular docking is to estimate the binding affinity between the ligand
and the target, which indicates how effectively a drug can inhibit or activate a
biological function.

In molecular docking, both the ligand and the receptor are modelled in three-
dimensional form. The ligand is placed into the binding site of the receptor in
different orientations, and each position is evaluated using scoring functions. These
scoring functions predict the stability and strength of the interaction based on factors
such as hydrogen bonding, hydrophobic interactions, electrostatic forces, and van der
Waals forces.

Molecular docking is generally divided into two main steps: search algorithm
and scoring function. The search algorithm explores possible conformations and
orientations of the ligand within the binding site, while the scoring function ranks
these conformations based on predicted binding energy. The best docking pose is the
one with the lowest binding energy and strongest interaction with the target
[Link] technique is widely used in drug discovery to screen large libraries of
compounds and identify potential drug candidates quickly and efficiently. It reduces
the need for extensive laboratory experiments by narrowing down the most promising
molecules for further testing. Molecular docking is especially useful in the
development of drugs for cancer, infectious diseases, and neurological disorders.

• Types of Molecular Docking

Molecular docking is classified based on the flexibility of the ligand (drug molecule)
and the receptor (target protein). The main types are

1. Rigid Docking

2. Flexible Ligand Docking

3. Flexible Receptor Docking

4. Induced Fit Docking

5. Ensemble Docking

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1. Rigid Docking

In rigid docking, both the ligand and the receptor are treated as completely rigid
structures. No flexibility is allowed in either molecule during docking. The ligand is
simply fitted into the fixed active site of the receptor. This method is fast and
computationally simple but less accurate because it does not reflect real biological
conditions.

2. Flexible Ligand Docking

In flexible docking, the ligand is allowed to rotate and change its shape while binding
to the receptor. The receptor remains rigid in most cases. This method provides more
realistic results compared to rigid docking because it considers different
conformations of the ligand.

3. Flexible Receptor Docking

In this method, the receptor protein is also allowed some flexibility during docking.
This is important because proteins can change shape when a ligand binds to them.
However, this method is more complex and requires higher computational power.

4. Induced Fit Docking

Induced fit docking allows flexibility in both the ligand and the receptor. It considers
that the protein binding site may adjust its shape to fit the ligand. This method is more
accurate and closely represents real biological interactions but is computationally
expensive.

5. Ensemble Docking

In ensemble docking, multiple different conformations of the receptor are used for
docking. This helps in considering protein flexibility without changing its structure
during simulation. It improves prediction accuracy and is widely used in modern drug
discovery.

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Fig no.3 Molecular Docking

1.5 Diabetes:

Diabetes mellitus is a chronic metabolic disorder characterized by elevated blood

glucose (hyperglycemia) due to defects in insulin secretion, insulin action, or both.
This imbalance leads to impaired metabolism of carbohydrates, fats, and proteins,
resulting in long-term damage and dysfunction of various organ especially eyes,
kidneys, nerves, heart, and blood vessels. Glucose is your body’s main energy source,
but it needs a hormone called Insulin to enter cells. In diabetes, either the body
doesn’t make enough insulin or can’t use it effectively. There are mainly two types of
diabetes: Type 1 diabetes, caused by autoimmune destruction of pancreatic β-cells
leading to absolute insulin deficiency, and Type 2 diabetes, which involves insulin
resistance and relative insulin deficiency. Type 2 diabetes is more common and is
often linked to obesity, sedentary lifestyle, and genetic predisposition. If not properly
managed diabetes can lead to severe complications such as neuropathy, nephropathy,
retinopathy, cardiovascular diseases, and delayed wound healing.

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Fig no.4 Image of types of diabetes

Types of Diabetes
1. Type 1 Diabetes (Insulin-Dependent Diabetes Mellitus) Caused by autoimmune
destruction of pancreatic β-cells, leading to absolute insulin deficiency. Usually
occurs in childhood or adolescence. Requires lifelong insulin therapy
2. Type 2 Diabetes (Non–Insulin-Dependent Diabetes Mellitus) Caused by insulin
resistance and a relative lack of insulin secretion. Common in adults, often linked
with obesity, sedentary lifestyle, and poor diet. Managed by diet control, exercise, oral
antidiabetic drugs, and sometimes insulin.

Signs and Symptoms

▪ Polyuria – frequent urination

▪ Polydipsia – excessive thirst

▪ Polyphagia – excessive hunger

▪ Unexplained weight loss (common in Type 1)

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▪ Fatigue and weakness

▪ Blurred vision

▪ Slow wound healing

▪ Numbness or tingling in hands and feet (neuropathy)

Fig no.5 Diabetes Symptoms, Risk Factor, Complication and Prevention

Antidiabetic Defenses:

1. Hormonal Defenses:
▪ Insulin: Secreted by pancreatic β-cells;promotes glucose uptake by
muscles and adipose tissue and inhibits hepatic glucose output.
▪ Glucagon: Secreted by α-cells; balances insulin by increasing blood
glucose during fasting.
▪ Incretins (GLP-1, GIP): Enhance insulin secretion after meals and
slow gastric emptying.
▪ Amylin: Co-secreted with insulin; helps regulate blood glucose spikes
after meals.

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1. Enzymatic / Metabolic Defenses:

▪ Glucokinase: Facilitates phosphorylation of glucose, aiding its utilization in

cells.
▪ Glycogen synthase: Converts glucose into glycogen for storage in liver and
muscle.
▪ AMP activated protein kinase (AMPK): Improves
insulin sensitivity and reduces hepatic glucose production
(metformin acts via this pathway).
▪ Glucose-6-phosphatase inhibition: Helps limit excess glucose release from
the liver.

2. Non-Enzymatic / Cellular Defenses:

▪ Insulin receptors and GLUT-4 transporters: Facilitate glucose entry

into muscle and fat cells.
▪ Adiponectin and leptin: Hormones that regulate glucose and lipid
metabolism.
▪ β-cell antioxidant system: Protects pancreatic cells from oxidative
damage caused by chronic high glucose levels.

➢ Prevention

• Maintain healthy body weight

• Balanced diet and regular exercise
• Avoid tobacco and excessive alcohol
Early detection through regular screening for high-risk individuals (family
history, obesity)

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1.6 Leucas Aspera : -

➢ Biological Source

Leucas aspera (Willd.) (Family: Lamiaceae) is a small, erect, annual herb known as
"Thumbai,"

➢ Geographical Source

Common weed found throughout South and Southeast Asia, with a primary
distribution across India, Bangladesh, Sri Lanka, the Philippines, Java, and Mauritius. It
thrives in tropical and subtropical regions, often found in cultivated fields, roadways, and
sunny areas.

➢ Macroscopic Characters
• Leaves: Opposite, simple, hairy, irregularly toothed margins
• Stem: Acutely quadrangular (square-shaped in cross-section)
• Flowers: Small, white, sessile, arrranges in dense terminal
• Bracts: 6 mm long, linear, acute, bristle-tipped, ciliate with long slender hairs.
• Odor: Aromatic
• Taste: Slightly bitter

➢ Microscopic Characters
• Stem Structure: Square-shaped in cross-section with four distinct ridges, containing a
single-layered epidermis
• Trichomes: Abundant non-glandular, multicellular (3-4 celled), uniseriate,
lignified hairs are present, alongside sessile glandular trichomes with multicellular heads.
• Cortex: Narrow parenchyma, with collenchymatous cells (3-8 rows) specifically
located under the ridges.
• Vascular Bundle: A ring of bicollateral vascular bundles connected by
interfascicular sclerenchyma
• Leaf Mesophyll: Displays distinct palisade and spongy parenchyma cells.

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➢ Chemical Constituents
• Flavonoids and Phenolic Compounds
• Triterpenoids and Triterpenoid Acids
• Flower Components:
• Alkaloids
• Essential oils
• Diterpenes and Glycosides
• Steroids (β-sitosterol)

➢ Pharmacological Actions
• Anti-Diabetic
• Anti-inflammatory
• Antimicrobial
• Antioxidant
• Hepatoprotective
• Anti-Cancer
• Used for skin problems, gout, rheumatism

➢ Mechanism of Action (Anti-Diabetic)

• Stimulating insulin secretion,
• Increasing insulin sensitivity
• Inhibiting glucose production or absorption
• Enhancing glucose excretion

➢ Uses
• Significant Blood Glucose Reduction
• Improved Glucose Tolerance
• Antioxidant Support
• Lipid Level Management:

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Figure no.6 Leucas Aspera Plant

2. AIM AND OBJECTIVE: -

AIM: -

To perform in-silico studies of chemical constituents of Leucas Aspera for antidiabetic

activity and comparison with metformin.

OBJECTIVE: -

The objective of the present study is to perform molecular docking of Metformin and of
chemical constituents of Leucas Aspera with the 5Y20 receptor to evaluate and compare their
binding affinity, molecular interactions, and potential inhibitory effects responsible for
antidiabetic activity. This computational approach will help in understanding the binding
mode, key amino acid residues involved in interaction, and structural requirements for
enhanced biological activity. The study of physicochemical properties, ADME and toxicity of
selected chemical constituents of Leucas Aspera by using computational software. The
outcome of the study will aid in identifying more potent lead molecule that may serve as
promising leads for further drug development against diabetes mellitus.

3. EXPERIMENT WORK: -

3.1 Downloading Software Program: -

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PYRX: - PyRx is an open-source virtual screening software used in Computer-Aided Drug

Design to perform molecular docking and analysed ligand–protein interactions. It provides a
user-friendly graphical interface that integrates tools such as AutoDock and AutoDock Vina
for docking simulations. PyRx allows researchers to import protein and ligand structures,
prepare molecules, and perform energy minimization before docking.

The software is widely used for virtual screening of large compound libraries to identify
potential drug candidates. It helps in predicting binding affinity and selecting the best ligand
based on docking scores. PyRx also supports visualization of molecular interactions, which
aids in understanding how a drug binds to its target receptor.

BIOVIA-DISCOVERY STUDIO: - BIOVIA Discovery Studio is widely used in Computer-

Aided Drug Design (CADD) to study interactions between ligands (drug molecules) and
biological targets such as proteins and enzymes. It allows researchers to visualize molecular
structures in three dimensions and analysed binding interactions at the atomic level.

The software includes various modules for molecular docking, pharmacophore modelling,
quantitative structure–activity relationship (QSAR) studies, and virtual screening.

AVAGADRO: - is an open-source molecular modelling and visualization tool used in

chemistry, pharmacology, and Computer-Aided Drug Design (CADD). It is designed to build,
edit, and visualize molecular structures in three-dimensional (3D) form.

CHEMSKETCH: - ACD/ChemSketch is a chemical drawing and molecular modeling

software developed by Advanced Chemistry Development (ACD/Labs). It is widely used in
chemistry, pharmacology, and Computer-Aided Drug Design (CADD) for drawing chemical
structures and predicting basic molecular properties.

3.2 Preparation of Ligand: -

Structure was drawn using chemsketch software and then the structure was cleaned by using
the clean structure tool and then structure was saved in the working folder as mol file. This
mole file was the accessed in Avogadro Software tool in which that the mol file is converted
to pdb format and then the structure was optimized by using the optimization tool and then
saved the optimized structure in the working directory as pdb file.

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Fig No.7- Structure of Ligand (Metformin)

Table No.1. Phytoconstituent of Leucas Aspera

Sr. Ligands IUPAC Name 2D Structure

No
1 Alpha 1- (3S,4S,5S,9R,10S,13R,14R,17R)-
Sitosterol 4,10,13-trimethyl-17-[(Z,2R)-5-
propan-2-ylhept-5-en-2-yl]-
2,3,4,5,6,9,11,12,14,15,16,17-
dodecahydro-1H-
cyclopenta[a]phenanthren-3-ol

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2 Baicalein 5,6,7-trihydroxy-2-phenylchromen-4-
one

3 Beta- (3S,8S,9S,10R,13R,14S,17R)-17-
sitosterol [(2R,5R)-5-ethyl-6-methylheptan-2-
yl]-10,13-dimethyl-
2,3,4,7,8,9,11,12,14,15,16,
17-dodecahydro-1H-cyclopenta[a]
phenanthren-3-ol

4 Campester : (3S,8S,9S,10R,13R,14S,17R)-17-
ol [(2R,5R)-5,6-dimethylheptan-2-yl]-
10,13-dimethyl-
2,3,4,7,8,9,11,12,14,15,16,
17-dodecahydro-1H-cyclopenta[a]
phenanthren-3-ol

5 Hexadecan hexadecane
e

6 Humulene (1E,4E)-2,6,6,9-
tetramethylcycloundeca-1,4,8-triene

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7 Leucaspero [(1S,5S,6S,8R)-8-acetyloxy-5-(1-
ne A acetyloxy-3-hydroxy-3-methylpent-4-
enyl)-1,5,6-trimethyl-2-oxo-4,6,7,8-
tetrahydro-3H-naphthalen-1-
yl]methyl acetate

8 Oleanolic (4aS,6aR,6aS,6bR,8aR,10S,12aR,14b
Acid S)-10-hydroxy-2,2,6a,6b,9,9,12a-
heptamethyl-
1,3,4,5,6,6a,7,8,8a,10,11,12,13,14b-
tetradecahydropicene-4a-carboxylic
acid

9 Oleic Acid octadec-9-enoic acid

10 Pentadecan pentadecane
e

11 Stigmaster (3S,8S,9S,10R,13R,14S,17R)-17-
ol [(E,2R,5S)-5-ethyl-6-methylhept
-3-en-2-yl]-10,13-dimethyl-
2,3,4,7,8,9,11,12,14,15,16,17
-dodecahydro-1H-cyclopenta
[a] phenanthren-3-ol

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12 Triterpenoi (4aS,6aR,6aS,6bR,8aR,9R,10S,12aR,
d 14bS)-10-hydroxy-2,2,6a,6b,9,12a-
hexamethyl-9-(sulfooxymethyl)-
1,3,4,5,6,6a,7,8,8a,10,11,12,13,14b-
tetradecahydropicene-4a-carboxylic
acid
13 Ursolic (1S,2R,4aS,6aR,6aS,6bR,8aR,10S,12
Acid aR,14bS)-10-hydroxy-
1,2,6a,6b,9,9,12a-heptamethyl-
2,3,4,5,6,6a,7,8,8a,10,11,12,13,14b-
tetradecahydro-1H-picene-4a-
carboxylic acid

14 Metformin 1-[(diaminomethylidene)amino]-N,N-
(Standard) dimethylmethanimidamide
hydrochloride

3.3 Preparation of Receptor: -

• The download PDB structure (5Y20) was opened in Discovery Studio Visualizer.
• All water molecule, heteroatoms, and co-crystallized ligand were removed to clean
the receptor.
• Polar hydrogen atom were added to stabilized the protein structure.
• The active site residues were defined based on the known Diabetes binding pocket.
• The final receptor file was saved in PDBQT format using AutoDock tools for
compatibility with docking software.

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Fig no.8 3ED Structure of receptor 5Y20

3.4 Physicochemical Properties: -

Lipinski rule was used to assess the physiochemical properties of all the selected ligands and
to predict their drug like properties, and the Swiss ADME as used to computer SMILE
structure of each compound.

3.5 ADME Studies: -ADME (Absorption, Distribution, Metabolism and Excretion) studies
are indeed crucial in drug development to assess how a drug behaves the body. Swiss ADME
software was used to determine these properties of each ligand.

3.6 Druglikeness :-

SwissADME evaluates drug-likeness by applying rules such as Lipinski, Ghose, Veber, Egan,
and Muegge, along with physicochemical properties like molecular weight, lipophilicity, and
polarity, to predict whether a compound is suitable for oral drug development.

3.7 Toxicity Study: -

It allows users to input a compound (via name, SMILES, or structure) and predicts over 60
toxicity endpoints, including acute toxicity (LD₅₀), organ toxicity (like hepatotoxicity and
neurotoxicity), carcinogenicity, mutagenicity, immunotoxicity, and clinical toxicity.

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The platform uses multiple predictive models (about 61) based on approaches such as
fragment analysis, pharmacophore modelling, and machine learning algorithms (e.g.,
Random Forest and neural networks) to generate results with confidence scores.

3.8 Molecular Docking: -

To perform molecular docking using PyRx:-

• Load protein and ligand files

• Perform ligand energy minimization
• Convert protein to macromolecule (PDBQT)
• Convert ligand to ligand format (PDBQT)
• Open Vina Wizard
• Select macromolecule and ligand
• Set grid box parameters
• Run docking using AutoDock Vina
• Analysed binding energy and poses
• Save/export docking results

4. RESULTS AND DISCUSSION: -

1. Physicochemical properties: -

The physicochemical properties of the compound were studied to predict the

pharmacokinetics of the drugs, using Lipinski’s Rule of Five is a set of guidelines used to
evaluate the drug-likeness of a compound, particularly its potential for good oral
bioavailability. It states that a molecule is more likely to be orally active if it has a molecular
weight of 500 Daltons or less, a lipophilicity value (Log P) not greater than 5, no more than 5
hydrogen bond donors, and no more than 10 hydrogen bond acceptors.

Table No.2. Physicochemical Properties of Ligands

Sr. Ligands No. of No. of No. of Molar Molecular TPSA

rotatable H- H-
No refractivity Weight
bonds bond bond
(g/mol)
accept
donors

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1 Alpha1- 2 2 3 36.93 126.16 91.49

Sitosterol

2 Beta Sitosterol 6 1 1 133.23 414.71 20.23

3 Baicalein 1 5 3 73.99 270.24 90.90

4 Campesterol 5 1 1 128.42 400.68 20.23

5 Hexadecane 5 1 1 137.56 426.72 137.56

6 Humulene 0 0 0 70.42 204.35 70.42

7 Leucasperone A 5 1 1 137.56 426.72 137.56

8 Oleanolic Acid 1 3 2 136.65 456.70 57.53

9 Oleic Acid 15 2 1 89.94 282.46 37.30

10 Pentadacane 12 0 0 74.22 212.41 0.00

11 Stigmasterol 5 1 1 132.75 412.69 20.23

12 Triterpenoid 4 3 7 148.27 552.76 129.51

13 Ursolic Acid 1 3 2 136.91 456.70 57.53

14 Metrformin 2 2 3 36.93 129.16 91.49

2. ADME Properties: -

ADME data predict using the Swiss ADME online web server database of phytocompounds.

Table No.3. Pharmacokinetics Properties of Ligands

Sr. Ligands GI BBB P-gp CYP1A2 CYP2C19 Log Kp

absorption
No permeant substrate inhibitor inhibitor (cm/s)

1 Alpha1- HIGH NO NO NO NO -7.99

Sitosterol

2 Beta Sitosterol LOW NO NO NO NO -2.20

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3 Baicalein HIGH NO NO NO NO -5.70

4 Campesterol LOW NO NO NO NO -2.50

5 Hexadecane LOW NO NO YES NO -1.80

6 Humulene LOW NO NO NO NO -4.32

7 Leucasperone LOW NO NO NO NO -2.49

A

8 Oleanolic Acid LOW NO NO NO NO -3.77

9 Oleic Acid HIGH NO NO YES NO -2.60

10 Pentadecane LOW NO NO YES NO -2.10

11 Stigmasterol LOW NO NO NO NO -2.74

12 Triterpenoid LOW NO YES NO NO -5.50

13 Ursolic Acid LOW NO NO YES NO -3.87

14 Metformin HIGH NO NO NO NO -7.99

3. Druglikeness :-

The drug-likeness capability of phytoconstituents can be predict using Lipinski, Ghose,

Veber, Egan, and Muegge rules which are based on certain physicochemical parameters.

Table No.4. Drug likeness Properties of Ligands

Sr. Ligands Lipinski Ghose Veber Egan Muegge Bioavailability

No score

1 Alpha1- YES NO YES YES NO 0.55

Sitosterol

2 Beta-Sitosterol YES NO YES NO NO 0.55

3 Baicalein YES YES YES YES YES 0.56

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4 Hexadecane YES NO NO NO NO 0.55

5 Humulene YES YES YES YES NO 0.55

6 Leucasperone YES NO YES NO NO 0.55

7 Oleanolic Acid YES NO YES NO NO 0.85

8 Oleic Acid YES NO NO NO NO 0.85

9 Pentadecane YES NO NO NO NO 0.55

10 Stigmasterol YES NO YES NO NO 0.55

11 Triterpenoid NO NO YES NO NO 0.56

12 Ursolic Acid YES NO YES NO NO 0.85

13 Campesterol YES NO YES NO NO 0.55

14 Metformin YES NO YES YES NO 0.55

4. Toxicity Study :-

Toxicity prediction is the important step in drug development and design. High demand for
computational predictive model to evaluate the potent toxic effects of drugs. In silico study
toxicity of drug is evaluating by Protox 3.0 online database.

Table No.5. Toxicity Study of Ligand

Sr. Ligands Predicted Predicted Carcinog Hepato Immuno Nephro

No Toxicity LD50 enicity toxicity toxicity toxicity

class (mg/kg)

1 Alpha1- 4 2000 INACTIVE INACTIVE ACTIVE INACTIVE

Sitosterol

2 Beta-Sitosterol 4 890 INACTIVE INACTIVE ACTIVE INACTIVE

3 Baicalein 5 3919 ACTIVE INACTIVE INACTIVE ACTIVE

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4 Campesterol 4 890 INACTIVE INACTIVE ACTIVE INACTIVE

5 Hexadecane 3 750 INACTIVE INACTIVE INACTIVE INACTIVE

6 Humulene 5 3650 INACTIVE INACTIVE INACTIVE ACTIVE

7 Leucasperone A 4 452 INACTIVE INACTIVE ACTIVE ACTIVE

8 Oleanolic Acid 4 2000 ACTIVE ACTIVE ACTIVE INACTIVE

9 Oleic Acid 2 48 INACTIVE INACTIVE ACTIVE INACTIVE

10 Pentadecane 3 750 ACTIVE INACTIVE INACTIVE INACTIVE

11 Stigmasterol 4 890 INACTIVE INACTIVE ACTIVE INACTIVE

12 Triterpenoid 6 6000 INACTIVE INACTIVE INACTIVE ACTIVE

13 Ursolic Acid 4 20000 ACTIVE ACTIVE ACTIVE INACTIVE

14 Metformin 4 680 INACTIVE INACTIVE INACTIVE INACTIVE

5. Binding affinity of Ligands :-

Table No.6. Binding affinity of phytoconstituents of Leucas Aspera with 5Y20 receptor

Sr. Ligands Binding affinity

No
1 Alpha1- Sitosterol -7.9
2 Beta-sitosterol -5.7
3 Baicalein -8.7
4 Campesterol -7.3
5 Hexadecane -4.1
6 Humulene -5.8
7 Leucasperone -5.7
8 Oleanolic Acid -9.0
9 Oleic Acid -6.0
10 Pentadecane -5.8

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11 Stigmasterol -7.8
12 Triterpenoid -7.4
13 Ursolic Acid -7.3
14 Metformin -5.0

6. 2D Structure of Phytoconstituents of Leucas Aspera: -

Table no.7. 2D Structures of Phytoconstituents of Leucas Aspera

Sr. LIGAND 2D STRUCTURE

No

1. ALPHA 1-
STIGMASTEROL

2. BETA SITOSTEROL

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3. BAICALEIN

4. CAMPESTEROL

5. HEXADECANE

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6. HUMULENE

7. LEUCASPERONE

8. OLEANOLIC ACID

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9. OLEIC ACID

10. PENTADECANE

11. STIGMASTEROL

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12. TRITERPENOID

13. URSOLIC ACID

14. METFORMIN

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5. CONCLUSION: -

The chemical constituent Alpha1-Sitosteroll have more binding affinity with receptor 5Y20
for antidiabetic activity. The ADMET studies indicates it was safer than metformin and
easily absorbed from GIT and cannot crossed BBB. Therefore, this molecule explores for
further development of new, potent antidiabetic agent with minimum side effects.

6. REFERENCES: -

1. Singh, H. et al. (2018). Drug Discovery and Development: An Overview. In:

Pharmaceutical Medicine and Translational Clinical Research. Elsevier.

2. Rang, H.P. & Hill, R.G. (2013). Drug Development. In: Drug Discovery and
Development (Second Edition). Elsevier. Available at:

3. Drug Discovery and Development: An Overview (ScienceDirect)

[Link]
4. Ece, A. (2023). Computer-Aided Drug Design. BMC Chemistry (ScienceDirect Topics).

[Link]
science/computer-aided-drug-design

5. Computational Biology and Chemistry (2026). Structure-based drug design; molecular

docking and QSAR.

[Link]

6. European Journal of Pharmaceutical Sciences (2023). CADD, AI and ML in drug

discovery: A comprehensive review.

[Link]

7. Kitchen, D. B., Decornez, H., Furr, J. R., & Bajorath, J. (2004). Docking and scoring in
virtual screening for drug discovery: methods and applications.

[Link]

8. Nabuurs, S. B., Wagener, M., & de Vlieg, J. (2007). A flexible approach to induced.

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[Link]

9. Chandrasekar, S. B., Bhanumathy, M., Pawar, A. T., & Somasundaram, T. (2010).

Phytopharmacology of Ficus religiosa. Pharmacognosy Reviews, 4(8), 195-199.
[Link]
10. Devanesan, E., Anand, V., Kumar, P. S., Vinayagamoorthy, P., & Basavaraju, P.
(2018). Phytochemistry and Pharmacology of Ficus religiosa. Systematic
ReviewinPharmacy,9(1),45-48. [Link]
and [Link]

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