“MOLECULAR DOCKING, SYNTHESIS AND CHARACTERIZATION OF

NOVEL N10-SUBSTITUTED ACRIDONE DERIVATIVES FOR THEIR IN-

VITRO CORRELATION OF LIPOPHILIC AND CYTOTOXIC ACTIVITY.”
SYNOPSIS FOR REGISTRATION OF M PHARM DISSERTATION
Submitted to
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

BENGALAURU, KARNATAKA.

By
SHIVARANJANI V S
M Pharm
Department of Pharmaceutical
Chemistry, Government College of
Pharmacy, Bengaluru, Karnataka –
560027.

Under the guidance

of Dr. SATHISH NK
Associate Professor,
Department of Pharmaceutical
Chemistry,
Government College of
Pharmacy, Bengaluru, Karnataka
- 560027.
 RAJIV GANDHI UNIVERSITY OF HEALTH
SCIENCES, BENGALURU, KARNATAKA
ANNEXURE-II
PROFORMA FOR REGISTRATION OF SUBJECT FOR
M PHARM DISSERTATION

1 Name of the Candidate Shivaranjani V S

D/o Siddalingaiah
Address Vadagere village,
Kannamangala
post,
doddabalapura
taluk, Bangalore
rural district-
561203

2 Name of the Institute Government College of Pharmacy,

#2, Kalinga Rao Road,
Subbaiah Circle,
Bengaluru - 560027

3 Course of study and subject Master of Pharmacy

(Pharmaceutical Chemistry)

4 Date of admission to course 03/02/2023

Title of the topic

5 “MOLECULAR DOCKING, SYNTHESIS AND
CHARACTERIZATION OF NOVEL N10-SUBSTITUTED
ACRIDONE DERIVATIVES FOR THEIR IN-VITRO
CORRELATION OF LIPOPHILIC AND CYTOTOXIC
ACTIVITY.”
6 Brief Resume of the Intended Work

6.1: Need for the Study

According to the World Health Organization, cancer is the second leading cause of death globally,
accounting for an estimated 9.6 million deaths, or one in six deaths, in 2018 (1).
Breast cancer is the second leading cause of cancer deaths among women.
The development of breast cancer is a multi-step process involving multiple cell types, and its prevention
remains challenging in the world. Early diagnosis of breast cancer is one of the best approaches to prevent
this disease.
It is a metastatic cancer and can commonly transfer to distant organs such as the bone, liver, lung and
brain, which mainly accounts for its incurability. Early diagnosis of the disease can lead to a good
prognosis and a high survival rate (2).
MCF-7 is a stable epithelioid cell line originally obtained from the pleural. effusion of a female patient
with metastatic breast cancer whose disease responded to hormone therapy. These cells have substantial
amount of estrogen receptor.
It is a human breast cancer cell line with estrogen, progesterone and glucocorticoid receptors. It is derived
from pleural effusion of a 69 -year-old Caucasian metastatic breast cancer (adenocarcinoma) in 1970.
MCF-7 cells are useful for in-vitro breast studies (3).

Lipophilicity is one of the most important parameters to be considered especially for anti-cancer drugs
as it plays a major role and responsible for permeation and accumulation of the drugs inside the cancer
cell. Increased Lipophilicity can be correlated with increased biological activity, poorer aqueous
solubility, more rapid metabolism and elimination, increased rate of skin penetration, increased
plasma protein binding (PPB), faster onset of action and occasionally shorter duration of action.
The main aim of the present study is to examine molecular docking, cytotoxicity and lipophilicity of a
series of novel N10-substituted Acridone derivatives determined experimentally by the UV spectroscopic
method and calculated using Software, as well as to find the correlation between Lipophilicity and
cytotoxicity (4).
6.2 REVIEW OF LITERATURE
An extensive literature survey reveals that acridone and its derivatives were reported to possess a
diverse biological activity.
The following documents were found on such studies:
 Mohammed Aarjane et. al., (2021) synthesized a series of novel acridone derivatives from
isoxazoline via 1,3-dipolar cycloaddition reaction between a variety of aryl nitrile oxides
and N- allyl acridones. The synthesized compounds were tested for their anti- bacterial
potency against four pathogenic bacteria, the two compounds 2a and 2k exhibited
antibacterial activity against E. coli and P. putida. The in silico molecular docking results
supported the antibacterial activity (5).

2a R1, R2, R3 = H
2k R2 = N(CH3)2 R3 = H
 Mayur YC et. al., (2021) studied the drug synergy of novel drug combinations involving
TMZ and novel Acridone derivatives (AC2, AC7 and AC26) using in silico and in vitro
methods in the treatment of Drug Resistance GLIOMA. The drug synergy revealed that
the combination of TMZ and AC26 was found to show synergistic effect on the drug
resistant Glioma cell lines(T-98) as well as drug sensitive Glioma cell lines (U-87) (6).
 Jiayong Huang et. al., (2021) synthesized a new coumarin acridone Fluorescence Probe
S via Ullmann coupling and Friedel crafts reaction, probe S exhibited high selectivity and
sensitivity for Fe3+ and it can penetrate into living organs for in vivo imaging. The Fe3+
within living cells and organs (7).
 Renfei Liu et. al., (2020) synthesized acridone derivatives with electron rich triphenyl
amine functionalized at the amino position and the photo physical properties were
investigated. The crystal structural analysis reveals that the acridone moiety tends to form π
– π stacking with the assistant of the pending phenyl rings to form C-H- - - O hydrogen
bonds with the carbonyl of acridone, which employed a good strategy in the design of
acridone based crystalline materials (8).

 Sathish NK et. al., (2019) synthesized a series of novel N10 – substituted 2 – nitro
acridones, the compounds exhibited strong DNA binding ability through intercalation using
CT-DNA. The DNA binding properties of the compounds was studied by monitoring the
changes in the UV Visible absorption spectra of acridones upon the addition of CT-DNA
 and the lipophilicity was determined using the software ALOGPS. The relationship
between Lipophilicity and binding ability showed poor correlation (9).

 Bing Shu et. al., (2018) synthesized a new series of acridone derivatives which targeted
the c-myc promoter i-motif. one of the derivative B19 selectively bound to i-motif without
binding to the G-quadraplex and duplex DNA, and down regulated c-myc gene
transcription which resulted ultimately in cancer cell apoptosis (10).
 B19

 Murahari M et. al., (2017) designed and synthesized 2,4-dimethyl acridones and were
characterized, the compounds were screened for anti-cancer and MDR modulation
activity against breast cancer sensitive MCF7 and resistant MCF7/ADR cell lines. 12e
and 12f showed better cytotoxicity profiles and the compounds showed favorable
interactions with P-gp by docking studies (11).

 Amareswararao M et. al.,(2016) synthesized nitric oxide donating derivatives by

linking nitric oxide donating group to acridone moiety and were evaluated for their
cytotoxic
 potentials in comparison with reference drugs Doxorubicin against human colon cancer
cell lines by using Sulphorhodamine-B assay. Compound 15 exhibited good cytotoxic
activity and correlations were observed when cytotoxic data of these molecules was
compared with nitrite production (12).

Compound 15
 Maryam Mohammadi-Khanaposhtani et. al., (2015) synthesized a series of novel
acridone linked to 1,2,3-triazole and evaluated in vitro for their acetylcholinesterase and
butyrylcholinestrase inhibitory activities. Among the synthesized compounds, 10-((1-(4-
chlorobenzyl)-1H-1,2,3-triazol-4-yl) methyl)-2-methoxyacridin-9(10H)-one, 9g depicted
the most potent anti-AChE activity (IC50 = 7.31 μM). Also, docking study confirmed the
results obtained through in vitro experiments and predicted possible binding
conformation(13).

9g
 Maryam Mohammadi-Khanaposhtani et. al., (2015) synthesized novel 9(10H)-acridone-
1,2,3-triazole hybrids and evaluated for cytotoxic and apoptosis inducing agents. The
preliminary in vitro anti proliferative activity test against MCF-7, T-47D and MDA-MB-
231cells employing MTT assay was examined. The compound 8c with IC 50 value of
11±4.8 µMwas more potent than etoposide against MCF-7 cells (14).
 Compound 8c
 Mehdi a Beniddir et. al., (2014) Synthesized a new acridone alkaloids, Chlorospermines
were isolated from the stem bark of Glycosmis chlorosperma by means of bio guided
isolation using an invitro enzyme assay against DYRK1A and their selectivity profile was
evaluated against a panel of various kinases, and molecular docking calculations provided
structural details for the interaction between these compounds and DYRK1A (15).

 Prasad VVSR et. al., (2011) synthesized 2,4-dichloro N10 substituted acridones as cytotoxic
agents, they were evaluated for their in vitro cytotoxic effects against drug sensitive and
resistant cancer cells. The acridones reacted with duplex DNA by intercalation and
possessed higher affinity to GC than AT base pairs of DNA. Compounds 14, 15 and 16
possessed higher cytotoxicity among the derivatives (16).
 Sathish NK et. al., (2010) synthesized 1,3-diacetoxy acridones and screened them for
theiranti-cancer activity against Enelich Ascies Carcinoma (EAC) using in vitro and
in vivo models. The compound 11 showed significantly high anti-cancer activity
against EAC in comparison with Vincristine as standard (17).

Compound 11
 Sathish NK et. al., (2010) synthesized 1,3-dimethyl acridone derivatives with different
alkyl side chain substituted at N10 position with highly basic amino groups at the
terminal end of the alkyl chain. All the molecules were screened for cytotoxicity
against Human Breast Adeno Carcinoma and Human Proteolytic Leukemia cell lines by
DNA binding studies. The compound 14 having Piperazine butyl side chain exhibited
potent cytotoxic activity against MCF-7 cell lines and DNA intercalating properties (18).

Compound 14

 Mayur YC et. al., (2009) synthesized a series of 2-fluoro N10 substituted acridone
derivatives with varying alkyl side chain (propyl/butyl) and a tertiary amino group at
the alkyl terminal and were screened against cancer cell lines. The compound 11
bearing tricyclic ring with a butyl methyl piperazine sidechain possessed the highest
binding affinity(19).
 Compound 11
 Sathish NK et. al., (2008) synthesized and investigated in vitro cytotoxicity against human
breast adenocarcinoma (MCF-7) and human promyelocytic leukemia (HL – 60) cell lines
fora series of novel substituted acridones. The compound 12 bearing diacetoxy acridone
with tertiary amines group at the terminal end of the alkyl side chains had strong inhibiting
activity against MCF-7 and HL-60 cell lines, which may be associated with their DNA
binding capacity. The examination of the relationship between Lipophilicity and
cytotoxicity properties showed a poor correlation (20).

Compound 12

 Mayur YC et. al., (2008) synthesized chloro acridone analogs as cytotoxic agents, in
vitro cytotoxic effects have been examined against Human Promyelocyclic leukemia
sensitive cell lines (HL-60), including multidrug cross resistance of two main (P-gp and
MPR)
 phenotype sublines vincristine resistant (HT/60/VINC) and doxorubicin resistant
(HL-60/DX). compound 4 showed very good activity against sensitive and
resistant call lines (21).

Compound 4

6.3 Objective of the study

 Molecular docking studies using software AutoDock, Discoverystudii.

 Synthesis of some novel N10-substituted acridone derivatives.

 Characterization of synthesized derivatives using physical data and analytical methods.

 Calculation of Lipophilicity using UV Spectrometer or Chromatography Method and software

ALOGP

 Study of cytotoxic effect.

 Correlation of Lipophilicity and cytotoxicity.

 6.4 SCHEME

X=Electron withdrawing group

R1= R2 = R3 =R4= H / F /electron withdrawing group.

Y = different substituted secondary amine.

 6.5 Synthesis of Acridone Derivatives
PROCEDURE
 Condensation of o-Chloro benzoic acid with substituted aniline in the presence of potassium
carbonate, isoamyl alcohol and copper powder for 6 to 8 hours yields substituted diphenyl
amine- 2-caboxylic acid.
 This upon refluxing with polyphosphoric acid at 1000C for about 3 hours results in cyclization
to give substituted Acridone.
 Later, N-alkylation will be carried out by stirring the acridone analogs with alkyl chain followed
by nucleophilic substitution at N10 position using different amines to give various N10-substituted
acridone derivatives.
 The synthesized derivatives will be identified and characterized by TLC, melting point, IR,
NMR and MS.
 6.6 BIOLOGICAL

ACTIVITY

LIPOPHILICITY

Lipophilicity is a fundamental molecular property used in quantitative structure activity relationship

analysis and plays a crucial role in the drug design, in relation to their behavior in aqueous and
organic phase in natural, technical and pharmacological processes.
This parameter has been used in connection with Blood Brain Barrier distribution or intestinal drug
absorption and is modeled by partition of solute between n-octanol and water (log p).
It has also been shown for anticancer compounds that Lipophilicity can be regarded as one of the
main properties correlating with cytotoxicity.
For this reason, a quantitative assessment of Lipophilicity is an important tool in QSAR studies.
The log p determination will be done by UV spectroscopic method/chromatographic method. The log p
values of the synthesized compounds will be calculated based on the areas of absorbance determined in
the aqueous and n-octanol phases that are directly proportional to the concentration according to the
Beers-lamberts law.

CYTOTOXICITY
Cytotoxicity is defined as the toxicity caused due to the action of chemotherapeutic agents on living
cells. The anti-cancer activity of the samples will be determined by the MTT assay which is used to
measure cellular metabolic activity as an indicator of cell viability, proliferation and cytotoxicity. This
colorimetric assay is based on the reduction of a yellow tetrazolium salt(3-(4,5-dimethylthiazol-2-yl)-
2,5-diphenyltetrazolium bromide or MTT) to purple formazan crystals by metabolically active
cells(22).
7 MATERIAL AND METHODS

7.1 Sources of Data:

1. The preliminary data that was required for the experimental study will be obtained from the
Library, Government college of Pharmacy, Bengaluru, Karnataka.
2. Library and information center, Rajiv Gandhi University of Health Sciences,
Bengaluru, Karnataka.
3. https://www.rguhs.ac.in/digitallibrary/Helinet%20about%20us.html#
4. https://www.sciencedirect.com
5. https://pubmed.ncbi.nlm.nih.gov/
6. https://scholar.google.com/
7. https://www.cancer.net/
8. https://www.springer.com/journal/11094

7.2 Method of collection of data

The experimental data will be obtained by experimenting in the laboratories of Department of
Pharmaceutical Chemistry, Government College of Pharmacy, Bengaluru.

7.3 Does the study require any investigation to be conducted on animals? If so, please describe
in brief.
-NO-

7.4 Has ethical clearance been obtained from your institute in case of 7.3?
Not Applicable
8 REFERENCES

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17;7:18-20p.
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and preventions of breast cancer. International journal of biological sciences. 2017;13(11):1387.
3. Horwitz KB, Costlow ME, McGuire WL. MCF-7: a human breast cancer cell line with estrogen,
androgen, progesterone, and glucocorticoid receptors. Steroids. 1975 Dec 1;26(6):785-95.

4. https://emerypharma.com/blog/drug-lipophilicity-and-absorption-a-continuous-
challenge- toward-the-goal-of-drug-discovery/
5. Aarjane M, Slassi S, Ghaleb A, Tazi B, Amine A. Synthesis, biological evaluation, molecular
docking and in silico ADMET screening studies of novel isoxazoline derivatives from acridone.
Arabian Journal of Chemistry. 2021 Apr 1;14(4):103057.
6. Chakravarty M, Ganguli P, Murahari M, Sarkar RR, Peters GJ, Mayur YC. Study of
Combinatorial Drug Synergy of Novel Acridone Derivatives With Temozolomide Using in-
silico and in-vitro Methods in the Treatment of Drug-Resistant Glioma. Frontiers in
oncology. 2021 Mar 15;11:640.
7. Huang J, Yan Z, Qiu P, Mo Y, Cao Q, Li Q, Huo L, Zhao L. A New Coumarin-Acridone
Compound as a Fluorescence Probe for Fe3+ and Its Application in Living Cells and
Zebrafish. Molecules. 2021 Jan;26(8):2115.
8. Liu R, Zhu G, Zhang G. N-Substitution of acridone with electron-donating groups: crystal
packing, intramolecular charge transfer and tuneable aggregation induced emission. RSC
Advances. 2020;10(12):7092-8.
9. Chaithra KR, Sathish NK, Channabasavaraj KP. Synthesis and DNA-binding studies of novel
2- Nitro acridones as anti-cancer agents. International Journal of Research i
10. Shu B, Cao J, Kuang G, Qiu J, Zhang M, Zhang Y, Wang M, Li X, Kang S, Ou TM, Tan JH.
Syntheses and evaluation of new acridone derivatives for selective binding of oncogene c-
myc promoter i-motifs in gene transcriptional regulation. Chemical Communications.
2018;54(16):2036-9.
11. Murahari M, Kharkar PS, Lonikar N, Mayur YC. Design, synthesis, biological evaluation,
molecular docking and QSAR studies of 2, 4-dimethylacridones as anticancer agents.
European journal of medicinal chemistry. 2017 Apr 21;130:154-70.
12. Prasad VR, Reddy GD, Kathmann I, Amareswararao M, Peters GJ. Nitric oxide releasing
acridone carboxamide derivatives as reverters of doxorubicin resistance in MCF7/Dx
cancer cells. Bioorganic chemistry. 2016 Feb 1;64:51-8.
13. Mohammadi-Khanaposhtani M, Saeedi M, Zafarghandi NS, Mahdavi M, Sabourian R,
Razkenari EK, Alinezhad H, Khanavi M, Foroumadi A, Shafiee A, Akbarzadeh T. Potent
acetylcholinesterase inhibitors: Design, synthesis, biological evaluation, and docking study
of acridone linked to 1, 2, 3-triazole derivatives. European journal of medicinal chemistry.
2015 Mar 6;92:799-806.
14. Mohammadi-Khanaposhtani M, Safavi M, Sabourian R, Mahdavi M, Pordeli M, Saeedi M,
Ardestani SK, Foroumadi A, Shafiee A, Akbarzadeh T. Design, synthesis, in vitro cytotoxic
activity evaluation, and apoptosis-induction study of new 9 (10 H)-acridinone-1, 2, 3-
triazoles. Molecular diversity. 2015 Nov;19(4):787-95.
15. Beniddir MA, Le Borgne E, Iorga BI, Loaë c N, Lozach O, Meijer L, Awang K, Litaudon
M. Acridone alkaloids from Glycosmis chlorosperma as DYRK1A inhibitors. Journal of
natural products. 2014 May 23;77(5):1117-22.
16. Babu YR, Bhagavanraju M, Reddy GD, Peters GJ, Prasad VV. Design and synthesis of
quinazolinone tagged acridones as cytotoxic agents and their effects on EGFR tyrosine
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17. Sathish NK, Raghavendra NM. In-vitro and in-vivo antitumor activity of 1, 3-diacetoxy
acridones against Ehrlich ascites Carcinoma. Asian Journal of Pharmaceutical and
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18. Sathish NK, GopKumar P, Prasad VR, Kumar SS, Mayur YC. Synthesis, chemical
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binding studies. Medicinal chemistry research. 2010 Sep;19(7):674-89.
19. Mayur YC, Peters GJ, Lemos C, Kathmann I, Prasad VV. Synthesis of 2‐Fluoro N10‐
Substituted Acridones and Their Cytotoxicity Studies in Sensitive and Resistant Cancer Cell
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Pharmaceutical and Medicinal Chemistry. 2009 Nov;342(11):640-50.
20. Sathish NK, Rajendra Prasad VV, Raghavendra NM, Shanta Kumar SM, Mayur YC.
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21. Prasad VR, Rao JV, Giri RS, Sathish NK, Kumar SS, Mayur YC. Chloro acridone derivatives
as cytotoxic agents active on multidrug-resistant cell lines and their duplex DNA complex
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9 Signature of the candidate

10 Remarks of the guide

Name and designation of;
11 Dr. Sathish NK
11.1 GUIDE Associate professor
Dept. of Pharmaceutical
Chemistry Govt. College of
Pharmacy Bengaluru-560027

11.2 SIGNATURE

11.3 HEAD OF THE DEPARTMENT Dr. Chandrasekhar Javali

Professor and Head
Dept. of Pharmaceutical
Chemistry Govt. College of
Pharmacy Bengaluru-560027

11.4 SIGNATURE

12 12.1 Remarks of the chairman and

the Principal

12.2 PRINCIPAL Dr. Kalaskar gurunath

Principal,
Govt. college of Pharmacy,
Bengaluru, Karnataka –
560027

SIGNATURE