Anti-HIV, Antitubercular and Antibacterial Activities of Novel 3-(Substituted Quinazolinylamino)-2-phenyl quinazolin-4(3H)ones

Article abstract of DOI:10.14233/ajchem.2020.22280

In the present study, we have synthesized a series of novel 2-phenyl-3-(substituted quinazolinylamino)quinazolin-4(3H)-ones by the reaction of 3-(substituted)-2-hydrazinoquinazoline-4(3H)-ones with 2-phenyl-3,1-benzoxazin-4-one. The starting material 3-(substituted)-2-hydrazino-quinazolin-4(3H)-ones were synthesized from various primary amines. All the synthesized compounds were screened for their antitubercular, anti-HIV and antibacterial activity against different Gram-positive and Gram-negative strains by agar dilution method. Among the test compounds, 3-(4-nitrophenyl)-2-(4-oxo-2-phenylquinazolin-3(4H)-ylamino)quinazolin-4(3H)-one (BQZ6) and 3-(4-chlorophenyl)-2-(4-oxo-2-phenylquinazolin-3(4H)-ylamino)quinazolin-4(3H)-one (BQZ7) shown most potent antibacterial activity against E. coli, P. aeruginosa and S. aureus with the MIC of 3 μg/mL. The compound BQZ7 exhibited the antitubercular activity with the MIC of 25 μg/mL and anti-HIV activity with the MIC of 35.4 μg/mL against HIV1 and HIV2 and offers potential lead for further optimization and development to new antitubercular and anti-HIV agents. The results from this study confirm that the synthesized and biologically evaluated quinazolines showed promising antimicrobial, antitubercular and anti-HIV activities and are new scaffolds for antimicrobial activity.

Full text of DOI:10.14233/ajchem.2020.22280

Asian Journal of Chemistry; Vol. 32, No. 2 (2020), 281-286
A
SIAN
J
OURNAL OF HEMISTRY
C
https://doi.org/10.14233/ajchem.2020.22280
Anti-HIV, Antitubercular and Antibacterial Activities of Novel
3-(Substituted Quinazolinylamino)-2-phenyl quinazolin-4(3H)ones
1
2
1,*
M.T. SULTHANA , K. CHITRA and V. ALAGARSAMY
1Medicinal Chemistry Research Laboratory, MNR College of Pharmacy, Sangareddy-502294, India
2Faculty of Pharmacy, Sri Ramachandra Institute of Higher Education and Research (Deemed to be University), Porur, Chennai-600116, India
*Corresponding author: Fax: +91 8455 230555; Tel: +91 8455 230690; E-mail: drvalagarsamy@gmail.com
Received: 19 July 2019;
Accepted: 2 September 2019;
Published online: 30 December 2019;
AJC-19713
In the present study, we have synthesized a series of novel 2-phenyl-3-(substituted quinazolinylamino)quinazolin-4(3H)-ones by the
reaction of 3-(substituted)-2-hydrazinoquinazoline-4(3H)-ones with 2-phenyl-3,1-benzoxazin-4-one. The starting material 3-(substituted)-
2-hydrazino-quinazolin-4(3H)-ones were synthesized from various primary amines. All the synthesized compounds were screened for
their antitubercular, anti-HIV and antibacterial activity against different Gram-positive and Gram-negative strains by agar dilution method.
Among the test compounds, 3-(4-nitrophenyl)-2-(4-oxo-2-phenylquinazolin-3(4H)-ylamino)quinazolin-4(3H)-one (BQZ6) and 3-(4-
chlorophenyl)-2-(4-oxo-2-phenylquinazolin-3(4H)-ylamino)quinazolin-4(3H)-one (BQZ7) shown most potent antibacterial activity against
E. coli, P. aeruginosa and S. aureus with the MIC of 3 µg/mL. The compound BQZ7 exhibited the antitubercular activity with the MIC
of 25 µg/mL and anti-HIV activity with the MIC of 35.4 µg/mL against HIV1 and HIV2 and offers potential lead for further optimization
and development to new antitubercular and anti-HIV agents. The results from this study confirm that the synthesized and biologically
evaluated quinazolines showed promising antimicrobial, antitubercular and anti-HIV activities and are new scaffolds for antimicrobial
activity.
Keywords: Quinazolines, Antibacterial activity, Antitubercular activity, Anti-HIV activity.
resistant tuberculosis (XDR-TB) strains. It clearly highlights
the urgent need to develop novel “druggable” molecules for
the co-infection treatment and strains of MDR-TB and XDR-
TB.
Recent year’s quinazolines and condensed quinazolines
gained much attention to medicinal chemists and pharmacologist
due to their potential druggable behaviour [5]. Among that
antimicrobial activity of 2,3-disubstituted quinazolines are
encouraging for further development. From recent literature
it is revealed that the 2,3-disubstituted quinazoline nucleus
showed significant antimicrobial activity [5,6].
Quinazolines and bisquinazolines have been explored for
various medicinal chemistry properties due to their widespread
pharmacological activities including antineoplastic, antimyco-
bacterial, antibacterial, antifungal, antiviral and antimalarial.
In recent literature, many quinazoline derived compounds were
prepared and screened for its antimicrobial activity. The current
use of these agents in bacterial and viral infections have led to
INTRODUCTION
Tuberculosis (TB) is an opportunistic infection that occurs
more often or more severe in people with weakened immune
systems than in people with healthy immune systems. Human
immunodeficiency virus (HIV) weakens the immune system,
increasing the risk of tuberculosis in people with HIV. Co-infection
with tuberculosis and HIV poses a tremendous challenge to
tuberculosis control, particularly in resource-limited treatment
option. In 2015, it was estimated 10.4 million cases of tuberculosis
disease, among that 1.2 million (11 %) people living with HIV,
there were an estimated 456,000 deaths from HIV-associated
tuberculosis [1-4]. Although exhaustive efforts to prevent and
treat tuberculosis is taken the problem still continues due to
multi-drug-resistant (MDR-TB) to isoniazid, rifampicin,
quinolones and aminoglycosides. Recently tuberculosis threat
has an additional challenge with the emergence of both multi-
drug-resistant tuberculosis (MDR-TB) and extensively drug-
This is an open access journal, and articles are distributed under the terms of the Attribution 4.0 International (CC BY 4.0) License. This
license lets others distribute, remix, tweak, and build upon your work, even commercially, as long as they credit the author for the original
creation. You must give appropriate credit, provide a link to the license, and indicate if changes were made.

282 Sulthana et al.
Asian J. Chem.
the development of novel anti-HIV, antibacterial and antituber-
cular agents [7-15]. In present study, using the quinazoline
scaffold we have developed 3-(substituted)-2-(4-oxo-2-phenyl-
quinazolin-3(4H)-ylamino)quinazolin-4(3H)-ones by incorpo-
rating the phenyl group at the C-2 position and quinazoline
moiety with desired substituents at N-3 position of quinazoline
as potent anti-HIV, antitubercular and antibacterial agents based
on pharmacophore approach. In this approach a molecule was
created by merging two quinazolines called as bisquinazolines.
Active site of targets may be addressed by this pharmacophore,
which increase the opportunity for selectivity. In addition it
also expected to reduce the undesirable side effects. The title
compounds were synthesized by a facile synthetic methods
with the good yields and studied for their anti-HIV activity in
MT-4 cells against replication of HIV-1 (III B) and HIV-2 (ROD);
antitubercular activity against M. tuberculosis H37RV by 10 fold
serial dilutions of each test compound/drug and the antibacterial
activity against selected Gram-positive and negative bacteria
by Agar dilution method.
hydrochloric acid. The solid obtained was filtered, washed with
water, dried and recrystallized from ethanol.Yield: 86 %; m.p.:
281-282 °C; IR (KBr, νmax, cm–1): 3355 (NH), 3062 (Ar-CH),
1729 (C=O), 1607 (C=C), 1520 & 1346 (NO2), 1251 (C=S);
1H NMR (CDCl3) δ: 6.73-6.75 (m, 2H, Ar-H), 7.26 (d, J = 7.5
Hz, 1H, Ar-H), 7.52-7.54 (m, 2H, Ar-H), 7.71 (d, J = 2.0 Hz,
1H, Ar-H),7.95 (d, J = 7.5 Hz, 1H, Ar-H), 8.01 (d, J = 2.0 Hz,
1H, Ar-H), 8.09 (m, 1H, Ar-H),9.16 (brs, 1H, CSNH); MS
(m/z): 299 [M+]; Anal. calcd. (%) for C14H9N3O3S: C, 56.18;
H, 3.03; N, 14.04. Found (%): C, 56.31; H, 3.02; N, 14.00.
(Adopting this procedure other 3-substituted-2-thioxo-2,3-
dihydro-quinazolin-4-ones are prepared).
Synthesis of 2-methylthio-3-(4-nitrophenyl)quinazolin-
4-one (4): The solution of compound 3 (0.01 mol) in alcoholic
sodium hydroxide solution (25 mL) transferred in to conical
flask. To this mixture dimethyl sulphate (0.01 mol) was added
drop wise with stirring and stirring was continued for 1 h after
complete addition of dimethyl sulphate. The reaction mixture
was then poured into ice cold water and mixed well. The resul-
tant solid obtained was filtered, washed with water and dried.
The crude solid obtained was recrystallized from ethanol.Yield:
91 %; m.p.: 181-182 °C; IR (KBr, νmax, cm–1): 3059 (Ar-CH),
2957 (CH3-CH), 1715 (C=O), 1668 (C=N), 1612 (C=C), 1534
& 1326 (NO2), 672 (C-S-C); 1H NMR (CDCl3) δ: 2.20 (s, 3H,
SCH3), 7.08-7.10 (m, 2H, Ar-H), 7.26 (d, J = 7.5 Hz, 1H, Ar-
H), 7.52-7.54 (m, 2H, Ar-H), 7.71 (d, J = 2.0 Hz, 1H, Ar-H),
7.90 (d, J = 7.5 Hz, 1H, Ar-H), 7.92 (d, J = 2.0 Hz, 1H, Ar-H);
MS (m/z): 313 [M+]; Anal. calcd. (%) for C15H11N3O3S: C,
57.50; H, 3.54; N, 13.41. Found (%): C, 57.67; H, 3.55; N,
13.38. (Adopting this procedure other 3-substituted-2-methyl-
sulfanyl-quinazolin-4-ones are prepared).
EXPERIMENTAL
In open capillaries melting points were measured on a
Thomas Hoover melting point apparatus (Thomas Hoover,
USA) and are uncorrected. Using potassium bromide disks the
IR spectra (ν, cm–1) were recorded on Bruker FT-IR spectro-
meter (Bruker, USA). Using Bruker FT NMR spectrometer
(Bruker, USA) the 1H NMR spectra were recorded in CDCl3 at
300 MHz. The chemical shifts are reported (δ, ppm) using
tetramethylsilane (TMS) as an internal standard. Using fast
atom bombardment (FAB positive) mass spectra were obtained
on a JEOL-SX-102 instrument (JEOL, Japan). PerkinElmer
2400 CHN analyzer (Perkin Elmer, USA) was used to perform
elemental analysis and values were within the acceptable limits
of the calculated values ( 0.4 %). Readymade silica gel plates
(Merck, Norway) are used to monitor the progress of the reaction.
All chemicals and reagents used in the synthesis were obtained
from Merck, SD fine chemicals, Aldrich (USA), Lancaster
(USA), or Spectrochem (India) and were used without further
purification.
Synthesis of 3-(4-nitrophenyl)-2-thioxo-2,3-dihydro
quinazolin-4-one (3): A solution of 4-nitro aniline (0.02 mol)
in dimethyl sulfoxide (10 mL) was stirred vigorously. To this
carbon disulphide (1.6 mL) and aqueous 20 M sodium hydro-
xide (1.2 mL) was added drop wise and stirring was continued
for 0.5 h. Dimethyl sulphate (0.02 mol) was added to the
reaction mixture and the reaction condition was kept in freezing
mixture for 2 h. The resultant reaction mixture was transferred
into ice cold water and stirred well. The product obtained was
filtered, washed with water, dried and recrystallized from
ethanol.
Synthesis of 2-hydrazino-3-(4-nitrophenyl)quinazolin-
4-one (5): The compound 4 (0.01 mol) was dissolved in ethanol
(25 mL). To this mixture 99 % hydrazine hydrate (0.1 mol)
and anhydrous potassium carbonate (100 mg) was added and
refluxed for 36 h. The reaction mixture was cooled and poured
into ice cold water. The solid so obtained was filtered, washed
with water, dried and recrystallized using ethanol to afford
the compound 5. Yield: 83 %; m.p.: 200-201 °C; IR (KBr,
ν
max, cm–1): 3371 & 3286 (NH), 3035 (Ar-CH), 1748 (C=O),
1
1650 (C=N), 1621 (C=C), 1519 & 1353 (NO2); H NMR
(CDCl3) δ: 5.72 (brs, 1H, Ar-NH), 6.17 (brs, 2H, NH2), 6.73-
6.75 (m, 2H, Ar-H), 6.94 (d, J = 7.5 Hz, 1H, Ar-H), 7.42-7.44
(m, 2H, Ar-H), 7.61 (d, J = 2.0 Hz, 1H, Ar-H), 7.95 (d, J = 7.5
Hz, 1H, Ar-H), 8.18 (d, J = 2.0 Hz, 1H, Ar-H); MS (m/z): 297
[M+]; Anal. calcd. (%) for C14H11N5O3: C, 56.56; H, 3.73; N,
23.56. Found (%): C, 56.45; H, 3.75; N, 23.64. (Adopting this
procedure other 2-hydrazino-3-substituted-quinazolin-4-ones
are prepared).
Synthesis of 2-phenyl-1, 3-benzoxazin-4-one: 14.05 g
of benzoyl chloride (0.2 mol) was added to a solution of 13.7
g of anthranilic acid (0.1 mol) dissolved in 60 mL pyridine.
For a period of 30 min the reaction mixture was stirred and
treated with 15 mL of NaHCO3 (5 %). The products separated
were crystallized from alcohol.Yield: 81 %; m.p.: 120-121 °C;
IR (KBr, νmax, cm–1): 3025 (Ar-CH), 1685 (C=O), 1620 (C=N),
1627 (C=C), 1029 (Cyclic C-O-C); 1H NMR (CDCl3) δ: 6.81-
6.84 (m, 3H, Ar-H), 6.95 (d, J = 7.5 Hz, 2H), 7.02-7.04 (m,
To the above prepared N-(4-nitrophenyl)-methyl dithiocar-
bamic acid (0.02 mol), methyl anthranilate (0.02 mol) solution
in ethanol (20 mL) was added. To the above reaction mixture
anhydrous potassium carbonate (100 mg) was added and refluxed
for 23 h. The reaction mixture was further cooled by using
ice. The resulting solid separated by filtration. The product 3
obtained was purified by dissolving in 10 % alcoholic sodium
hydroxide solution and re-precipitated by treating with dilute

Vol. 32, No. 2 (2020)
Biological Studies of Novel 3-(Substituted Quinazolinylamino)-2-phenyl quinazolin-4(3H)ones 283
3H, Ar-H), 7.51 (d, J = 2.0 Hz, 1H); MS (m/z): 223 [M+];Anal.
calcd. (%) for C14H9NO2: C, 75.33; H, 4.06; N, 6.27. Found
(%): C, 75.11; H, 4.07; N, 6.29.
Synthesis of 3-(substituted quinazolinyl amino)-2-
phenyl quinazolin-4(3H)ones (BQZ1-BQZ10): A mixture
of 2-hydrazino-3-substituted-3H-quinazolin-4-one (5) (0.01
mol) and 2-phenyl-3,1-benzoxazin-4-one (0.01 mol) were
fused at 150 °C and the solid obtained and recrystallized to
afford the title compounds (BQZ1-BQZ10).
3-Ethyl-2-(4-oxo-2-phenylquinazolin-3(4H)-ylamino)-
quinazolin-4(3H)-one (BQZ1): Yield: 86 %; m.p.: 228-229
°C; IR (KBr, νmax, cm–1): 3372 (NH), 3057 (Ar-CH), 2989
(CH3-CH), 1703 (C=O), 1655 (C=N), 1610 (C=C); 1H NMR
(CDCl3) δ: 1.52-1.83 (t, 3H, CH3), 2.46-2.69 (q, 2H, CH2),
5.64 (s, 1H, Ar-NH), 7.10-7.95 (m, 13H, Ar-H); MS (m/z):
409 [M+]; Anal. calcd. For C24H19N5O2: C, 70.40; H, 4.68; N,
17.1043. Found (%): C, 70.19; H, 4.70; N, 17.16.
3-Allyl-2-(4-oxo-2-phenylquinazolin-3(4H)-ylamino)-
quinazolin-4(3H)-one (BQZ2): Yield: 80 %; m.p.: 225-226
°C; IR (KBr, νmax, cm–1): 3304 (NH), 3028 (Ar-CH), 2963
(CH2-CH), 1702 (C=O), 1649 (C=N), 1607 (C=C); 1H NMR
(CDCl3) δ: 1.64-1.88 (d, 2H, CH2CH=CH2), 4.72-4.95 (d, 2H,
CH2CH=CH2), 5.37-5.51 (m, 1H, CH2CH=CH2), 5.62 (s, 1H,
Ar-NH), 7.04-8.16 (m, 13H,Ar-H); MS (m/z): 421 [M+];Anal.
calcd. (%) for C25H19N5O2: C, 71.25; H, 4.54; N, 16.62. Found
(%): C, 71.50; H, 4.52; N, 16.57.
2-(4-Oxo-2-phenylquinazolin-3(4H)-ylamino)-3-
phenylquinazolin-4(3H)-one (BQZ3): Yield: 81 %; m.p.:
240-241 °C; IR (KBr, νmax, cm–1): 3258 (NH), 3045 (Ar-CH),
1727 (C=O), 1662 (C=N), 1609 (C=C); 1H NMR (CDCl3) δ:
5.53 (s, 1H, Ar-NH), 7.27-8.10 (m, 18H, Ar-H); MS (m/z):
457 [M+]; Anal. calcd. (%) for C28H19N5O2: C, 73.51; H, 4.19;
N, 15.31. Found (%): C, 73.74; H, 4.17; N, 15.26.
2-(4-Oxo-2-phenylquinazolin-3(4H)-ylamino)-3-
(pyridin-4-yl)quinazolin-4(3H)-one (BQZ4): Yield: 80 %;
m.p.: 251-252 °C; IR (KBr, νmax, cm–1): 3349 (NH), 3034 (Ar-
CH), 1730 (C=O), 1653 (C=N), 1616 (C=C); 1H NMR (CDCl3)
δ: 5.60 (s, 1H, Ar-NH), 6.91-8.24 (m, 17H, Ar-H); MS (m/z):
458 [M+]; Anal. calcd. (%) for C27H18N6O2: C, 70.73; H, 3.96;
N, 18.33. Found (%): C, 70.45; H, 3.97; N, 18.39.
2-(4-Oxo-2-phenylquinazolin-3(4H)-ylamino)-3-p-
tolylquinazolin-4(3H)-one (BQZ5):Yield: 83 %; m.p.: 232-
233 °C; IR (KBr, νmax, cm–1): 3394 (NH), 3051 (Ar-CH), 2952
(CH3-CH), 1735 (C=O), 1650 (C=N), 1623 (C=C); 1H NMR
(CDCl3) δ: 2.98 (s, 3H, CH3), 5.46 (s, 1H, Ar-NH), 7.13-8.25
(m, 17H, Ar-H); MS (m/z): 471 [M+]; Anal. calcd. (%) for
C29H21N5O2: C, 73.87; H, 4.49; N, 14.85. Found (%): C, 73.60;
H, 4.48; N, 14.91.
m.p.: 230-231 °C; IR (KBr, νmax, cm–1): 3343 (NH), 3039 (Ar-
CH), 1704 (C=O), 1646 (C=N), 1611 (C=C), 786 (C-Cl); 1H
NMR (CDCl3) δ: 5.61 (s, 1H,Ar-NH), 7.26-8.14 (m, 17H, Ar-
H); MS (m/z): 493 [M+2], 491 [M+]; Anal. calcd. (%) for
C28H18ClN5O2: C, 68.36; H, 3.69; N, 14.24. Found (%): C,
68.13; H, 3.70; N, 14.29.
3-(2-Chlorophenyl)-2-(4-oxo-2-phenylquinazolin-
3(4H)-ylamino)quinazolin-4(3H)-one (BQZ8): Yield: 78 %;
m.p.: 238-239 °C; IR (KBr, νmax, cm–1): 3286 (NH), 3053 (Ar-
CH), 1731 (C=O), 1668 (C=N), 1605 (C=C), 764 (C-Cl); 1H
NMR (CDCl3) δ: 5.42 (s, 1H,Ar-NH), 7.34-8.01 (m, 17H, Ar-
H); MS (m/z): 493 [M+2], 491 [M+]; Anal. calcd. For
C28H18ClN5O2: C, 68.36; H, 3.69; N, 14.24. Found (%): C,
68.19; H, 3.68; N, 14.28.
3-(2-Methoxyphenyl)-2-(4-oxo-2-phenylquinazolin-
3(4H)ylamino)quinazolin-4(3H)-one (BQZ9): Yield: 78 %;
m.p.: 243-244 °C; IR (KBr, νmax, cm–1): 3360 (NH), 3042 (Ar-
CH), 2943 (CH3-CH), 1719 (C=O), 1654 (C=N), 1628 (C=C),
1061 (C-O-C); 1H NMR (CDCl3) δ: 3.05 (s, 3H, OCH3), 5.58
(s, 1H,Ar-NH), 7.02-8.37 (m, 17H,Ar-H); MS (m/z): 487 [M+];
Anal. calcd. (%) for C29H21N5O3: C, 71.45; H, 4.34; N, 14.37.
Found (%): C, 71.19; H, 4.36; N, 14.42.
3-(4-Methoxyphenyl)-2-(4-oxo-2-phenylquinazolin-
3(4H)-ylamino)quinazolin-4(3H)-one (BQZ10): Yield: 81 %;
m.p.: 215-216 °C; IR (KBr, νmax, cm–1): 3337 (NH), 3020 (Ar-
CH), 2979 (CH3-CH), 1706 (C=O), 1651 (C=N), 1613 (C=C),
1051 (C-O-C); 1H NMR (CDCl3) δ: 3.40 (s, 3H, OCH3), 5.57
(s, 1H,Ar-NH), 7.29-8.13 (m, 17H,Ar-H); MS (m/z): 487 [M+];
Anal. calcd. (%) for C29H21N5O3: C, 71.45; H, 4.34; N, 14.37.
Found (%): C, 71.70; H, 4.33; N, 14.32.
Antitubercular activity: Into Middle brook 7H11 agar
slants 10 fold serial dilutions of each test compound/drug were
incorporated with OADC growth supplement. Fresh Middle
brook 7H11 agar slants with OADC growth supplement was
used to prepare inoculums of M. tuberculosis H37RV and adjus-
ted to 1 mg/mL in Tween 80 (0.05 % W/V) saline diluted to
10-2 to give a 107 cfu/mL concentrate approximately. Into 7H11
agar tubes containing 10 fold serial dilutions of drug per mL a
5 µL amount of bacterial suspension was spotted.At 37 °C the
tubes were incubated and after 28 days the final readings were
recorded. Control tubes with medium alone were incubated
with H37RV were used to compare tubes having the compounds.
Active concentration of test compound was taken as the concen-
tration at which complete inhibition of colonies occurred. The
minimum concentration of compound required to give complete
inhibition of bacterial growth was taken as MIC [16-18].Against
reference drug isoniazid, rifampicin and ethambutol the MIC
of the test compounds was compared.
3-(4-Nitrophenyl)-2-(4-oxo-2-phenylquinazolin-3(4H)-
ylamino)quinazolin-4(3H)-one (BQZ6): Yield: 80 %; m.p.:
241-242 °C; IR (KBr, νmax, cm–1): 3271 (NH), 3046 (Ar-CH),
1728 (C=O), 1664 (C=N), 1602 (C=C), 1540 & 1317 (NO2);
1H NMR (CDCl3) δ: 5.52 (s, 1H, Ar-NH), 7.15-8.08 (m, 17H,
Ar-H); MS (m/z): 502 [M+]; Anal. calcd. (%) for C28H18N6O4:
C, 66.93; H, 3.61; N, 16.73. Found (%): C, 67.12; H, 3.60; N,
16.75.
Anti-HIV activity: In MT-4 cells anti-HIV activity of the
compounds (BQZ1-BQZ10) were tested against replication
of HIV-1 (III B) and HIV-2 (ROD) [19]. The MT-4 cells were
grown in RPMI-1640 DM (Dutch modification) medium (Flow
laboratories, Irvine, Scotland), supplemented with 10 % (v/v)
heat inactivated fetal calf serum and 20 µg/mL gentamicin (E.
Merck, Darmstadt, Germany). From the culture supernatant
of HIV-1 infected MT-4 cell lines HIV-1 (III B) and HIV-2
(ROD) were obtained and the virus stocks were stored at -70 °C
until used. Microtiter plates was used to perform anti-HIV
3-(4-Chlorophenyl)-2-(4-oxo-2-phenylquinazolin-
3(4H)-ylamino)quinazolin-4(3H)-one (BQZ7):Yield: 81 %;

284 Sulthana et al.
Asian J. Chem.
assay by filled with 100 µL of medium and 25 µL volumes of
compounds in triplicate so as to allow simultaneous evaluation
of their effects on HIV and mock infected cells. 50 µL of HIV
at 100 CCID50 medium was added to either infected or mock
infected part of microtiter tray. At 37°C the cell cultures were
incubated in a humidified atmosphere of 5 % CO2 in air. By the
MTT method after 5 days of infection spectrophotometrically
examined the viability of mock and HIV infected cells. The
effective dose of compound achieving 50 % protection of MT-
4 cells against the cytopathic effect (Virus cause cell degeneration
or cell death, which can be seen by microscopical examination
of cultures. Cell degeneration is manifested by certain patho-
logical changes) of HIV (EC50) and the cytotoxic dose of comp-
ound, required to reduce the viability of normal uninfected
MT-4 cells by 50 % (CC50) were calculated.
Antibacterial activity: Agar dilution method was used
to evaluate antibacterial activity of compounds [20,21]. From
theAmerican type culture collection (ATCC), Rockville, USA
the standard strains were procured and the pathological strains
were procured from the department of microbiology, MNR
medical college, Sangareddy, India. The antibacterial activity
of the test analogs was screened against the following bacterial
strains: E. coli ATCC25922, P. vulgaris ATCC9484, S. typhimurium
ATCC 33068, K. pneumoniae ATCC 13883, P. aeruginosa ATCC
2853, B. subtilis ATCC 6051, S. aureus ATCC 25923, M. luteus
ATCC 10240, S. epidermidis ATCC 35984, S. albus ATCC 17900.
Muller-HintonAgar (Hi-media) plates (37 °C, 24 h) were used
for bacterial growth and the minimum inhibitory concentration
(MIC) was considered to be the lowest concentration that
completely inhibited the growth on agar plates, disregarding
a single colony or faint haze caused by the inoculums. Cipro-
floxacin was used as reference drug for comparison of MIC
of the test compounds.
mate, which was methylated with dimethyl sulfate to afford
the dithiocarbamic acid methyl ester, which on reflux with methyl
anthranilate (2) in ethanol yielded the desired 3-substituted-
2-thioxo-2,3-dihydro-1H-quinazolin-4-one (3)in a good yield.
The 3-substituted-2-methylsulfanyl-3H-quinazolin-4-one (4)
was obtained by dissolving compound (3) in 2 % alcoholic
sodium hydroxide solution and methylating with dimethyl
sulphate by stirring at room temperature. With hydrazine
hydrate nucleophilic displacement of methylthio group of (4)
was carried out in ethanol to afford 2-hydrazino-3-substituted-
3H-quinazolin-4-one (5). The title compounds 3-(substituted
quinazolinylamino)-2-phenyl quinazolin-4(3H)ones (BQZ1-
BQZ10) were obtained by the condensation of amino group
of 2-hydrazino-3-phenyl-3H-quinazolin-4-one (6) with 2-phenyl-
1,3-benzoxazin-4-one. The formation of title product is indicated
by the disappearance of peak due to NH2 of the starting material
1
in the IR and H NMR spectrum of the compounds BQZ1-
BQZ10. In the IR and 1H NMR spectrum of these peaks due
to thiosemicarbazides, carbonyl (C=O), NH and aryl groups
presence were confirmed. The mass spectra of test compounds
molecular ion peaks corresponding to their molecular formulae
were observed. Elemental (C, H, N) analysis satisfactorily
confirmed elemental composition and purity of the synthesized
compounds.
Antitubercular activity: The synthesized compounds
were screened for their in vitro antimycobacterial activity
against M. tuberculosis strain H37RV. The results are expressed
in terms of Minimum Inhibitory Concentration (MIC). The results
of antimycobacterial activity is depicted in Table-1, indicates
that the test compounds inhibited the growth of Mycobacterium
at the minimum microgram of 25 µg/mL concentration.
Anti-HIV activity:The results of anti-HIV activity (Table-1)
indicate that all compounds exhibited mild to moderate anti-
HIV activity; whereas compounds BQZ7 containing aryl ring
with electron withdrawing group exhibited anti-HIV activity
at 35.4 µg/mL concentration against HIV1 and HIV2. While
the test compounds with other substituent’s showed mild to
moderate anti-HIV activity against HIV1 and HIV2.
Antibacterial activity:Among the different substituent at
N-3 position of the quinazoline, aryl and heteroaryl substituents
RESULTS AND DISCUSSION
Synthetic route shown in Scheme-I outline the chemistry
part of the present work. The key intermediate 3-substituted-
2-thioxo-2,3-dihydro-1H-quinazolin-4-one (3) was obtained
by reacting primary amine (1) with carbon disulphide and sodium
hydroxide in dimethyl sulphoxide to give sodium dithiocarba-
TABLE-1
ANTITUBERCULAR, ANTI-HIV AND ANTIBACTERIAL ACTIVITY OF TITLE COMPOUNDS (BQZ1- BQZ10)
Test compounds and standard
Microorganism
BQZ1 BQZ2 BQZ3 BQZ4 BQZ5 BQZ6 BQZ7 BQZ8 BQZ9 BQZ10
STD1
STD2 STD3
M. tuberculosis
HIV-1
HIV-2
S. typhi
E. coli
B. subtilis
K. pneumonia
P. vulgaris
P. aeruginosa
S. aureus
M. luteus
S. epidermidis
S. albus
25
100
100
100
50
100
50
100
100
100
25
25
39.6
39.6
25
13
25
13
13
6
25
49.5
49.5
25
47.6
47.6
25
100
100
100
50
100
50
100
50
50
25
50
100
25
35.9
35.9
25
3
25
13
13
3
25
35.4
35.4
25
3
25
13
13
3
3
6
6
13
25
39.1
39.1
25
6
25
25
50
6
13
25
13
25
25
54.9
54.9
50
50
25
25
25
13
13
25
91.4
91.4
50
100
50
50
50
25
25
0.05
0.0012
0.000642
0.1
–
–
–
–
–
–
–
–
–
–
–
–
1.56
–
–
–
–
–
–
–
–
–
–
–
–
25
25
25
13
25
25
25
13
25
25
25
25
25
25
13
13
6
25
25
25
4
1
1
1
1
1
1
1
1
1
6
3
13
13
25
13
13
25
25
50
50
13
100
50
50
100
Antitubercular standard: STD1 - Isoniazid, STD2 - Rifampicin, STD3 - Ethambutol; Anti-HIV standard: STD1-AZT; Antibacterial standard:
STD1 - Ciprofloxacin.

Vol. 32, No. 2 (2020)
Biological Studies of Novel 3-(Substituted Quinazolinylamino)-2-phenyl quinazolin-4(3H)ones 285
S
R
NH C
DMSO
R
S-Na+
NH2 + CS2 + NaOH
(1)
(CH3)2SO4
O
O
OCH3
EtOH
OCH3
S
R
C
NH
+
S-Na+
NH2
NH R
C
S
NH
(2)
O
N
O
R
N
R
NaOH
N
SCH3
DMS/EtOH
S
N
(4)
H
EtOH
NH2NH2.H2O
(3)
O
N
O
O
R
N
NHNH2
Fused at 150 oC
N
(5)
O
N
BQZ-6 R =
CH2CH CH2 BQZ-7 R =
NO2
Cl
BQZ-1 R =
BQZ-2 R =
C2H5
R
N
O
NH
Cl
N
BQZ-3 R = C6H5
BQZ-4 R =
BQZ-8 R =
BQZ-9 R =
H3CO
N
N
(6)
BQZ-5 R =
CH3
BQZ-10 R =
OCH3
Scheme-I: Synthesis of quinazolinyl quinazolines (BQZ1-BQZ10)
exhibited better activity over the aliphatic and cyclic substi-
tuent. Compounds with electron withdrawing substituents like
chloro and nitro showed better activity over the unsubstituted
and electron donating substituents.Among the test compounds
3-(4-nitrophenyl)-2-(4-oxo-2-phenylquinazolin-3(4H)-yl-amino)-
quinazolin-4(3H)-one (BQZ6) and 3-(4-chlorophenyl)-2-(4-oxo-
2-phenylquinazolin-3(4H)-ylamino)quinazolin-4(3H)-one
(BQZ7) shown most potent activity againstE. coli, P. aeruginosa
and S. aureus with the MIC of 3 µg/mL. Compound BQZ7 and
BQZ6 emerged as the most active compounds of the series.
Conclusion
Synthesis of new series of 3-(substituted)-2-(4-oxo-2-phenyl-
quinazolin-3(4H)-ylamino)quinazolin-4(3H)-ones have been

286 Sulthana et al.
Asian J. Chem.
7. F.R. Pavan, P.I.S. Maia, S.R.A. Leite, V.M. Deflon, A.A. Batista, D.N.
Sato, S.G. Franzblau and C.Q.F. Leite, Eur. J. Med. Chem., 45, 1898
(2010);
https://doi.org/10.1016/j.ejmech.2010.01.028.
8. O. Güzel, N. Karali andA. Salman, Bioorg. Med. Chem., 16, 8976 (2008);
https://doi.org/10.1016/j.bmc.2008.08.050.
9. N. Karali, A. Gürsoy, F. Kandemirli, N. Shvets, F.B. Kaynak, S. Ozbey,
V. Kovalishyn and A. Dimoglo, Bioorg. Med. Chem., 15, 5888 (2007);
https://doi.org/10.1016/j.bmc.2007.05.063.
10. D. Sriram, P. Yogeeswari, R. Thirumurugan and R.K. Pavana, J. Med.
Chem., 49, 3448 (2006);
described. The title compounds exhibited significant antibac-
terial activity against the different Gram-positive and Gram-
negative bacteria including M. tuberculosis; and they also
exhibited significant activity against HIV1 and HIV2 strains.
The substituents at 3-aminoquinazoline shown varied anti-
microbial activity, aryl substituents with electron withdrawing
group showed most potent and the aryl/allyl substituents
showed moderate activity and the alkyl, aryl substituents with
electron donating groups showed the least activity. Among
the test compounds, compounds 3-(4-nitrophenyl)-2-(4-oxo-
2-phenylquinazolin-3(4H)-ylamino)quinazolin-4(3H)-one
(BQZ6) and 3-(4-chlorophenyl)-2-(4-oxo-2-phenylquinazolin-
3(4H)-ylamino)quinazolin-4(3H)-one (BQZ7) shown most
potent activity against E. coli, P. aeruginosa and S. aureus with
the MIC of 3 µg/mL. The compound BQZ7 exhibited the anti-
tubercular activity at the minimum microgram of 25 µg/mL
and anti-HIV activity at 35.4 µg/mL against HIV1 and HIV2
and offers potential lead for further optimization and develop-
ment to new antitubercular and anti-HIV agents.
https://doi.org/10.1021/jm060339h.
11. D. Sriram, P. Yogeeswari, P. Dhakla, P. Senthilkumar, D. Banerjee and
T.H. Manjashetty, Bioorg. Med. Chem. Lett., 19, 1152 (2009);
https://doi.org/10.1016/j.bmcl.2008.12.088.
12. E. Saripinar, Y. Güzel, S. Patat, I. Yildirim, Y. Akçamur and A.S.
Dimoglo, Arzneimittelforschung, 46, 824 (1996).
13. B. Milczarska, H. Foks, J. Sokolowska, M. Janowiec, Z. Zwolska and
Z. Andrzejczyk, Acta Pol. Pharm., 56, 121 (1999).
14. G. Turan-Zitouni, A. Özdemir, Z. Asim Kaplancikli, K. Benkli, P.
Chevallet and G. Akalin, Eur. J. Med. Chem., 43, 981 (2008);
https://doi.org/10.1016/j.ejmech.2007.07.001.
15. S.A. Shiba, A.A. El-Khamry, M.E. Shaban and K.S. Atia, Pharmazie,
52, 189 (1997).
16. D. Sriram, P. Yogeeswari, J.S. Basha, D.R. Radha and V. Nagaraja,
Bioorg. Med. Chem., 13, 5774 (2005);
CONFLICT OF INTEREST
https://doi.org/10.1016/j.bmc.2005.05.063.
The authors declare that there is no conflict of interests
regarding the publication of this article.
17. P. Shanmugavelan, S. Nagarajan, M. Sathishkumar, A. Ponnuswamy,
P.Yogeeswari and D. Sriram, Bioorg. Med. Chem. Lett., 21, 7273 (2011);
https://doi.org/10.1016/j.bmcl.2011.10.048.
18. J. Kunes, J. Bazant, M. Pour, K. Waisser, M. Slosárek and J. Janota, Il
Farmaco, 55, 725 (2000);
https://doi.org/10.1016/S0014-827X(00)00100-2.
19. R. Pauwels, E. De Clercq, J. Desmyter, J. Balzarini, P. Goubau, P.
Herdewijn, H. Vanderhaeghe and M. Vandeputte, J. Virol. Methods,
16, 171 (1987);
REFERENCES
1. World Health Organization, Global Tuberculosis Report 2018, World
Health Organization: Geneva, Switzerland (2018).
2. T.T. Balcha, S. Skogmar, E. Sturegard, P. Björkman and N. Winqvist,
Glob. Health Action, 8, 27048 (2015);
https://doi.org/10.3402/gha.v8.27048.
https://doi.org/10.1016/0166-0934(87)90002-4.
3. M.J. Reid and N.S. Shah, Lancet Infect. Dis., 9, 173 (2009);
https://doi.org/10.1016/S1473-3099(09)70043-X.
20. A. Barry, Antibiotics in Laboratory Medicine, William and Wilkins:
Baltimore, MD, edn 5, vol. 1 (1991).
4. World Health Organization Tuberculosis, Available online: https://
www.who.int/tb/en/ (accessed on 17 March 2019).
21. S.N. Pandeya, D. Sriram, G. Nath and E. De Clercq, Il Farmaco, 54, 624
(1999);
5. V. Alagarsamy, K. Chitra, G. Saravanan, V.R. Solomon, M.T. Sulthana
and B. Narendhar, Eur. J. Med. Chem., 151, 628 (2018);
https://doi.org/10.1016/j.ejmech.2018.03.076.
https://doi.org/10.1016/S0014-827X(99)00075-0.
6. A. Hameed, M. Al-Rashida, M. Uroos, S.A. Ali, Arshia, M. Ishtiaq
and K.M. Khan, Expert Opin. Ther. Pat., 28, 281 (2018);
https://doi.org/10.1080/13543776.2018.1432596.