This document discusses potential herbal treatments for COVID-19 from Tulsi plant extracts. It summarizes the rationale, objectives, and introduction to SARS viruses. Molecular docking studies show good binding affinity of chicoric acid, rosmarinic acid, and caftaric acid from Tulsi with SARS-CoV-2 proteins. Formulations for herbal mouthwash, gargle, nasal spray, nasal gel and throat paint were developed using Tulsi and evaluated for stability and efficacy.

1. 1
ANTI-VIRAL HERBAL
PHYTOCONSTITUENTS
AND FORMULATION
OF TULSI (OCIMUM
SANCTUM) AGAINST
COVID-19.
Guided by,
Dr. Yamini D. Shah
Head of department of
Pharmaceutics
Submitted by : Kavish Shah
B. Pharm
LM College of pharmacy,
Ahmedabad.

2. Rationale
 At present there is no effective treatment that
could mitigate SARS-CoV-2. Available clinical
intervention for covid-19 is only limited to
support.
 Herbal medicine has a wide array chemical entity
with potential health benefits and cure including
antiviral activity which may be explored as
alternative treatment of COVID-19.
 Major content of medicinal plant are structural
analogue of surface protein of SARS-CoV-2
determined through molecular docking.
Objective
 To provide basic knowledge of the disease and risk associated with
patients.
 To promote the effective implementation of infection control measures in
order to development of herbal medicine using Tulsi plant.
 Safety measure of patients can be evaluated through parameter with
respect to herbal formulation.
2

3. Introduction to SARS viruses:
 SARS is known as severe acute respiratory
syndrome (SARS) fatal viral disease and it is
rapidly spreading to all over globe.
 According to the Centers for Disease Control and
Prevention (CDC), USA, SARS was recognized as a
global threat in March 2003.
 The disease was first seen in the city of Foshan in
Guangdong Province in China in 2002 and was
traced back to the ​Himalayan (masked) palm civet,
racoon dogs and the Chinese ferret-badger (1).
 All of these animals are sold in the live wild animal
markets in China. The civet cats were thought to be
the intermediaries who passed the SARS virus from
bats to humans.
Viruses
 A virus is defined as an obligate intracellular parasite.
They are only seen by electron microscope.
3

4. Classification scheme of H-CoV and other coronavirus
4

5. Newly emerging Human corona virus:
 Pathogenic human corona viruses emerged beginning with the discovery of SARS-CoV
in 2002.
 Presently there are now seven human coronaviruses (2).
1. HCoV-229E
2. HCoV-NL63
3. HCoV-OC43
4. HCoV-HKU1
5. SARS-CoV
6. MERS-CoV
7. SARS-CoV2
5

6. Clinical signs and symptoms
Patients with SARS show flu-like symptoms that will improve after the first week
(30% of patients) and 20-30% Patients will require special care. A history of SARS-
CoV-2 contact is often found in patients with immense infection. Initial symptoms
of SARS-CoV-2 patient include:
 fever
 rigors
 myalgia
 malaise
 cough
 pleuritic pain
 dyspnea
pneumonia-like symptoms Complications include, acute respiratory distress
syndrome (ARDS), acute cardiac injury, tachycardia, hypotension, acute renal
failure, encephalitis (3).
6

7. Origin of SARS-CoV-2
The International Committee on the Taxonomy of Viruses (ICTV) suggested that
SARS-CoV-2 belonging to the species of Severe acute respiratory syndrome
currently considers SARS-CoV-2 as sampled from a patient in Wuhan, China, on
26 December 2019 .
By January 2019, approximately eighteen countries were reported to be infected
with the virus emerged in China led to the declaration as “Public Health
Emergency of International Concern” by World Health Organization (WHO) on
Mar 11, 2020. As on July 21, 2021 there have been reported 19,06,71,330
confirmed cases with 40,98,758 fatality cases worldwide.
7

8. Structure of corona virus with spike
protein
 Corona viruses are single stranded positive RNA
viruses comprising of membrane enveloped
genome with neucleotides size range between 26-
32 kilobases. SARS-CoV-2 encode five structural
proteins
 Spike protein (S)
 Hemagglutinin esterase protein (HE)
 Membrane protein (M)
 Envelope protein (E)
 Neucleocapcid protein (N)
 S Protein, located outside of the virus is
glycoprotein.
 This glycoprotein gives typical shape to the virion.
Thus virion activates inside living body.
 These proteins act as cell surface receptor protein
for their penetration into the host cell and interact
with membrane protein.
8

9.  Hemagglutinin esterase (HE) proteins are surface proteins of SARS-CoV-2
which can be found in some beta- coronaviruses in order to strengthen
invading mechanism of that particular virus.
 Membrane proteins are glycosylated glycoproteins that are essential for
regeneration of virus cell.
 They are also necessary to fuse into host cell .
 E (Envelope proteins) are hydrophobic small proteins, covering whole
membrane. They allow attachment to the membrane of viruses and play a
key role in the combining of viral particles in the host cell.
 N (Neucleocapcid proteins) are also called as phosphoproteins that are able
to bind with viral genomic RNA. These proteins are important for structure,
replication and transcription mechanism of coronaviruses through
interactions with the viral RNA genome (4).
9

10. Classification Human CoVs Symptoms Receptor protein
α-coronavirus HCoV-229E Mild respiratory tract
infections
Human aminopeptidase N (CD13)
β-coronavirus HCoV-OC43 Mild respiratory tract
infections
9-O-acetylated sialic acid
β-coronavirus SARS-CoV Severe acute respiratory
infections, 10% mortality
rate
Angiotensin-converting enzyme 2
(ACE2)
α-coronavirus HCoV-NL63 Mild respiratory tract
infections
Angiotensin-converting enzyme 2
(ACE2)
β-coronavirus HCoV-HKU1 Pneumonia, lung
inflammation
9-O-acetylated sialic acid
β-coronavirus MERS-CoV Severe acute respiratory
infections, 37% mortality
rate
Dipeptidyl peptidase 4 (DPP4)
β-coronavirus SARS-CoV-2 Fever, cough, muscle pain,
weakness, respiratory
symptoms, shortness of
breath and death.
Angiotensin-converting enzyme 2
(ACE2) and transmembrane
protease serine 2 (TMPRSS2)
Human coronavirus and other coronavirus receptor binding site
10

11. SARS-CoV-2 virus entry in cell via ACE2 receptor and TMPRSS2
11

12. Anti-viral drugs against SARS-CoV-2
12
6

13.  Fusion inhibitor
1. Baricitinib
2.Umifenovir
3.Camostat mesylate
 Protease inhibitor
1.Lopinavir
2.Darunavir
3.Atazanavir
 Reverse transcription inhibitors
1.Remdesivir
2.Favipiravir (Avigan)
3.Ribavirin
 Neuraminidase inhibitors
1.oseltamivir
2. zanamivir
3.peramivir
 Chloroquine
 Hydroxychoroquine
Anti viral drugs against covid-19
Chloroquine tablets
Hydroxychloroquine tablets
13

14. Indian herbs against covid-19
 The traditional system of herbal medicine focused on cure of the disease and
promptly implementing infection control.
 Various potential Indian herbs
Liquorice Tulsi Turmeric
Giloy cinnamon Ashwagandha
14

15. Kalmegh Wheatgrass Neem
 Potential selected Indian herbs such as Ocimum sanctum (Tulsi), Glycyrrhiza
glabra (Liquorice), Curcuma domestica Vahl (Turmeric), Tinospora cordifolia
(Giloy), Withania somnifera (Ashwagandha), Cinnamon (Dalchini), Shoot of
Triticumaestivum Linn. (Wheatgrass), and Andrographis paniculata
(Kalmegh) with their antiviral properties to prevent infection.
 Potential of herbal plants for their biological effects as growth promotion,
immunostimulation, antistress, antibacterial, antifungal, antivirals, appetite
stimulators, and aphrodisiac .
15

16. Tulsi
 A recent study in India reported that Ocimum Sanctum (Tulsi) effective in the
prevention and management of COVID19.
 It is found in a tropical climate like Asia, Africa, Central, and South America.
 Basil leaves of ocimum sanctum contains Caffeic acid derivative
 chicoric acid (chicoric acid; also known as dicaffeoyltartaric acid, which is a
caffeic acid derivatized with tartaric acid)
 Rosmarinic acid
 Caftaric acid (in the order of most abundant to least; all derivatives of caffeic
acid)
 Rosmarinic acid was the main phenolic compound found in both leaves and
stems.
 Chicoric acid was not detected in sweet basil stems, although a small amount
was present in Thai basil stems (11).
16

17. Computer Assisted Drug Design
 The conventional approach to the drug design and development may take
plenty of resources such as cost, time and manpower with no guarantee to
afford the desired candidates with efficiency and potency.
 Therefore, the computer assisted drug design (CADD) have been preferred
over several years above the traditional approaches reducing the cost-burden
and improved results through biological science.
 CADD provide significant interactions of identified hits against their
biological targets to understand their mode of action.
 We have performed the virtual screening of 3 ligands Chicoric acid,
Rosmarinic acid and Caftaric acid from pubchem using AutoDock Vina
against RdRp(RNA Dependent RNA polymerase) and Membrane
protein using molecular docking docking in search of effective SARS CoV-2
inhibitors motivated by the potential of computational chemistry.
Experimental Section
17

18. Computational studies for analysis of membrane protein and RNA
dependent RNA polymerase
 The molecular docking has been performed against selected protein (PDB
ID: 7BV2 (RNA Dependent RNA polymerase and 6M0J (Membrane
protein) using AutoDock Vina to evaluate binding affinity of ligand and
interactions in the active site.
 This proteins and ligands in PDBQT format have been used for docking
process.
Caftaric acid Rosmarinic acid Chicoric acid
18

19. Compound no. Dataset Name Binding Energy
(Kcal/mol)
1. Chicoric acid(A) -8.6
2. Caftaric acid(B) -7.1
3. Rosmarinic acid(C) -8.2
Binding Energy of ligands with 7BV2 (RNA Dependent RNA polymerase
protein)
Compound no. Dataset Name Binding Energy
(Kcal/mol)
1. Chicoric acid(A) -7.6
2. Caftaric acid(B) -5.8
3. Rosmarinic acid(C) -6.0
Binding Energy of ligand with 6M0J (Membrane protein)
19

20. Compound
No.
Compound Name Binding Energy
(Kcal/mol)
1. Favipiravir -6.7
2. Penciclovir -7.2
3. Molnupiravir -10
4. Sofosbuvir -8.7
5. Remdesivir -9.2
20

21. 3D interaction of ligands A, B, C with 7BV2 (RdRp).
Chicoric acid Cafaric
acid
Rosmarinic acid
21

22. Chicoric acid Caftaric acid
Rosmarinic acid
3-D interaction of ligands A, B, C with membrane protein (6M0J).
22

23. Molecular Docking
The Pubchem website contains proteins and ligands. From pubchem we
downloaded 3 ligands namely chicoric acid (A), caftaric acid (B) and rosmarinic
acid (C). All Ligands have been optimized and converted into PDBQT format
using OpenBabel and further used for molecular dockings.
Molecular docking with three ligands was performed using the prepared protein
using Autodock vina to reveal the binding affinity and interactions of the
lingands with RdRp protein (PDB ID: 7BV2) and membrane protein (6M0J).
All the ligands were docked on the site using optimized grid box and ten poses
per ligand were generated. The docking interaction of hits have been visualized
and analyzed using Biovia Discovery studio 2020 (12).
23

24. Formulation Development
We have prepared herbal mouthwash, gargle, nasal spray, nasal gel and
throat paint using tulsi and further evaluates parameter for determine
stability and efficacy.
Gargle Mouthwash Throat paint
Evaluation parameters
 Colour and odour
 pH
 Stability studies
24

25. Nasal spray Nasal gel
Evaluation parameters
 Determination of pH
 Viscosity of solution
 Drug content uniformity
 Sterility
 Stability
 Drug content assay
 Spreadability
25

26. References
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Nature. 2020 Mar 12;579(7798):265–9.
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https://www.who.int/publications/i/item/9789240018440
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pathogenesis of SARS-CoV-2. BMJ. 2020;371:1–6.

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sanctum) and neem (Azadirachta indica) against SARS-CoV-2 Protein Targets.
Biol Eng Med Sci Reports. 2020;6(1):11–3.
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