Design, Synthesis, Molecular Docking, Invitro Anticancer and Antibacterial Evaluation of Novel Pyrazole Linked with Quinazoline Scaffolds

Introduction

One of the most common aberrant cell growths in the human body is cancer. Death rates from cancer exist. If cancer is detected in its early stages, it can be cured; if it is detected in its later stages, there are few possibilities that it can be cured.¹

Heterocyclic chemistry becomes more challenging for medicinal chemists. Heterocyclic compounds and their derivatives show various pharmacological and clinically active. Heterocyclic molecules have diversified action in various pharmacological and therapeutic activities.² The organic molecules are distributed in nature and play an essential role in regulating biological processes. Fused heterocyclic moieties like pyrazole and quinazoline nucleus are significant in the drug discovery process The heterocyclic compounds like pyrazole,³  quinazoline⁴ nucleus contain prominent electron withdrawing groups like nitrogen and oxygen atom. They have the potency to bind the receptor and show an agonist effect to receptor and it exhibit molecular inhibitory binding with biological systems. Due to their privilege and efficacy towards pharmacological significance. It has wide-linked properties of Pyrazole and quinazoline nucleus has a key role of inhibiting certain receptors like aurora kinase inhibitors ⁵ cyclin-dependent kinases (CDK) inhibitors,⁵ reticular activating system-neuroendocrine tumor (Ras-Net),⁵  DNA binding agent,⁵  epithelial growth factor receptor (EPGR)⁶ inhibitor, cyclo-oxygenase(COX),⁷   lipoxygenase(LOX) ⁷ inhibitors and tubulin inhibition polymerization ⁸ further active inhibitory receptor is represented in Figure1.⁹  It attach to different hetero molecules scaffolds published in the literature data for the potency compounds has with a wide range of therapeutic properties. Pyrazole¹⁰ and quinazoline¹¹ core moiety exhibit diverse pharmacological and therapeutic properties such as anticancer,¹² anti-mycobacterial,¹³ antidiabetic,¹⁴ anti-inflammatory,¹⁵ antimicrobial,¹⁶ anti-leishmanial,¹⁷ anti-hypertensive agents α-blocking,¹⁸ Neuroprotective agents¹⁸ as approved by USFDA drugs are shown in Figure2

Figure 1: Importance of inhibitory activity of Pyrazole and Quinazoline nucleus. Click here to View Figure

Figure 2: USFDA-approved drugs Pyrazole and Quinazoline nucleus. Click here to View Figure

Extremely efforts to develop a synthesis of pyrazole linked with quinazoline and its scaffolds. In our research, grateful to these pyrazole and quinazoline hybrids in continuation of our work towards biologically active molecules.¹⁹ In this research our aim to synthesized newly novel pyrazole and quinazoline hybrids via Schiff mechanisms and mannish base mechanism and its scaffolds (3a-l). Synthesized scaffolds are promising against cancer cell lines  MCF-7, PC-3, and HT-29 by using MTT assay methods. Antibacterial activity is screened by using the agar-diffusion method and insilico molecular docking ²⁰ is performed.

Materials and Methods

All the chemicals were purchased from SD Fine and AURA Laboratories Analytical research grades. Then further characterization by different analytical techniques like silica gel (TLC) Thin layer chromatography and IR spectroscopy (BRUKER), ¹H-NMR spectroscopy (400 MHz AvanceCore), ¹³C spectroscopy (Avance 400 MHz), Mass spectrometry (Thermo fisher scientific Orbitrap Exploris 120) and Melting point (LB-MPS8). Anti-cancer activity were performed by using MTT assay and antibacterial activity were performed by using bacterial strain and in silicomolecular docking are predicted.

Synthesis of N-(2-methyl-4-oxoquinazolin-3(4H)-yl)-3-oxo-3-(3,5-diphenyl-2H-pyrazol-1(5 H)-yl) propanamide.

The procedure consist of three steps

Step I: Synthesis of Chalcones

Acetophenone and different substituted benzaldehydes an add ethanol Stirr at magnetic stirrer at room temperature1hr by using Claisen– Schmidt condensation mechanism and the reaction was observed in TLC. The reaction crude product was filter and recrystallized with ethanol and finally obtained a chalcone moiety.

Step II: Synthesis of Quinazoline malonic dihydrazide

 Take Malonic di-hydrazide and add 2-methyl benzoxazine-4-one and add pyridine and refluxed for 3hr. In reaction mixture add crush ice or cold water neutralised with dil HCl. Filter the crude product and Precipitate were collected, dry the obtained product and it was recrystallized with methanol or ethanol.

Step III: Synthesis of Pyrazole Scaffolds (3a-3l)

Chalcone of step1 and malonic-dihydrozido quinazoline step2 was taken in RBF and add glacial acetic acid and add NaOH (40% 10 ml)  then reflux for  5hr. The reaction undergo cyclization schiff and mannish base mechanism formation of pyrazole and its scaffold (3a-l). The progress of the reaction mixture was monitored in TLC cooled with rinse water and add crushed ice. Filter the reaction and obtain solid residue, dry the residue, and recrystallized with ethanol.

Biological activity evaluation

In-vitro Anti-cancer activity

Anti-cancer activity were evaluated (3a-l) against three cancer cell line breast cancer (MCF7) and prostate cancer (PC-3) and human colon cancer (HT-29) by using MTT assay method ²¹, Trypsinization of the cells and trypan blue assay were performed to determine the viability of the cells in suspension. Trypsinization of the cells and trypan blue assay were performed to determine the viability of the cells in suspension. The cells were then counted using a hemocytometer (IMPROVED B.S.748 NEUBAUER by Rohem india, and a density of 5.0 X 10³ cells per well were seeded 100µL in culture medium in 96 well plates, and incubated Manufacturer Thermo scientific Model Co2 incubator model 370 manual for 12hrs at 37⁰C. Removed the existing medium, then filled each well with new media. Following that, plates were incubated for three hours at 37⁰C. Precipitations were produced at the end of the incubation time as a result of the MTT salt being reduced to chromophore-Formosan crystals by cells with metabolically active mitochondria. The optical density of crystals that had been dissolved in DMSO was calculated at 570 nm on a microplate reader using a FlexStation3 plate reader. The standard formula for calculating cell toxicity is (average OD*100/positive control)-100, and figures for the concentration of the test drug needed to hinder cell development by 50% (IC50) were utilized.

In-vitro Antibacterial activity

   Antibacterial activity was evaluated (3a-l) against gram-positive and gram-negative bacterial strains by using agar plate diffusion method.²² It is examined that the view a dose-dependent inhibition of bacterial growth.  

Insilico Molecular docking studies

The insilico molecular docking studies were predicted in Autodock vina PyRx 0.8.²³ This tool was analyze and predict the ligand and completely bound to the receptor. Viewed in 3D structure. The protein was downloaded from WWW.RCSBPDB.COM ⁽²³⁾ PDB: 5C5S (Human-myosin9b Rhogap), 6XXN (sting CTD complex), 3K46 (E.Coli beta glucurinadase) and finally docking visualization analysis was predicted in Discovery Studio and Molegro Molecular Viewer.

Spectral data of newly synthesized compound

3a: N-(2-methyl-4-oxoquinazolin-3(4H)-yl)-3-oxo-3-(3,5-diphenyl-2H-pyrazol-1(5 H)-yl) propanamide.

IR: 3413(-NH), 3377(-NH), 3134(-CH), 2989(-CH₃),2892(-CH),1725 (C=O), 1700 (C=O), 1555(C=N), 1295(C-N). ¹H NMR: 12.1 (1H, -NH), 11.33(1H, -NH), 8.4- 6.9(13H Ar), 6.35 (1H, -CH),  3.2 (1H, -CH₂), 2.42(3H, -CH₃). ¹³C NMR 127.4, 133.5, 12.4, 147.1, 120.9, 128.8, 164, 161, 19.9, 170.3, 44.5, 95.3, 61, 134, 128.7, 128.0, 128.7, 126.4 MASS (ESI) m/z: 466 [M+H]⁺

3b: N-(2-methyl-4-oxoquinazolin-3(4H)-yl)-3-oxo-3-(5-phenyl -3-p-tolyl-2H-pyrazol-1(5 H)-yl) propanamide.

IR: 3455 (-NH), 3302(-NH),  3102(-CH), 2957(-CH),1700 (C=O), 1670 (C=O), 1585(C=N), 1252(C-N).¹H NMR: 12.5 (1H, -NH,), 11.3(1H, -NH),  8.0- 6.9(13H Ar), 6.83 (1H, -CH),  3.42 (1H, -CH₂), 2.34 (3H, -CH₃).  1.90 (3H, -CH₃).  ¹³C NMR 127.4, 133.5, 12.4, 147.1, 120.9, 128.8, 164, 161, 19.9, 170.3, 44.5, 95.3, 61, 134, 128.7, 128.0, 128.7, 126.4, 19.9, 24.3 MASS (ESI) m/z: 478 [M+H]⁺

3c: 3-(3-(4-methoxyphenyl)-5-phenyl-2H –pyrazol-1(5 H)-yl-N-(2-methyl-4-oxoquinazolin-3(4H)-yl)-3-oxopropanamide.

IR: 3395 (-NH), 3355(-NH),  3144(-CH), 2980(-CH₃),2895(-CH),1715 (C=O), 1710 (C=O), 1545(C=N), 1285(C-N), 1245(C-O-C-OCH₃).¹H NMR: 11.9 (1H, -NH), 10.3 (1H, -NH),  8.0- 6.9 (13H Ar), 6.55 (1H, -CH),  3.4 (1H, -CH₂ ), 2.4 (3H, -CH₃).  2.33 (3H, -OCH₃). ¹³C NMR 127.4, 133.5, 12.4, 147.1, 120.9, 128.8, 164, 161, 19.9, 170.3, 44.5, 95.3, 61, 134, 128.7, 128.0, 128.7, 126.4, 159.9, 55.9 MASS (ESI) m/z: 496 [M+H]⁺.

3d: N-(2-methyl-4-oxoquinazolin-3(4H)-yl)-3-(3-(4-nitrophenyl)-5-phenyl-2 H-pyrazol-1(5 H)-yl)-3-oxopropanamide.

IR: 3395 (-NH), 3365 (-NH),  3133(-CH), 2945(-CH₃), 2900 (-CH),1710 (C=O), 1715 (C=O), 1590 (-NO₂), 1550 (C=N), 1290 (C-N), ¹H NMR: 12.0 (1H, -NH), 11.3 (1H, -NH),  8.0- 6.9 (13H Ar), 6.7 (1H, -CH),  3.0 (1H, -CH₂), 2.10 (3H, -CH₃) ¹³C NMR  127.4, 133.5, 12.4, 147.1, 120.9, 128.8, 164, 161, 19.9, 170.3, 44.5, 95.3, 61, 134, 128.7, 128.0, 128.7, 126.4, 127.3, 121.0 MASS (ESI) m/z: 511 [M+H]⁺.

3e: 3-(3-(4-chlorophenyl)-5-phenyl-2H –pyrazol-1(5 H)-yl-N-(2-methyl-4-oxoquinazolin-3(4H)-yl)-3-oxopropanamide

IR: 3415 (-NH), 3385 (-NH),  3145 (-CH), 2960(-CH₃),2890 (-CH),1710 (C=O), 1695 (C=O), 1570 (C=N), 1265 (C-N), 890 (C-Cl).¹H NMR: 12.0 (1H, -NH), 11.2 (1H, -NH),  8.0- 6.8(13H), 6.25(1H, -CH),  3.2 (1H, -CH₂), 2.2 (3H, -CH₃) ¹³C NMR 127.4, 133.5, 12.4, 147.1, 120.9, 128.8, 164, 161, 19.9, 170.3, 44.5, 95.3, 61, 134, 128.7, 128.0, 128.7, 126.4, 133.5, 128.8, 127.8 MASS (ESI) m/z: 500 [M+H]⁺.

3f: 3-(3-(3,4-dimethoxyphenyl)-5-phenyl-2H –pyrazol-1(5 H)-yl-N-(2-methyl-4-oxoquinazolin-3(4H)-yl)-3-oxopropanamide

IR: 3403 (-NH), 3333(-NH),  3160 (-CH), 2955 (-CH₃), 2890 (-CH),1710 (C=O), 1705 (C=O), 1535 (C=N), 1295 (C-N), 1255 (C-O-C-OCH₃).¹H NMR: 12.2 (1H, -NH), 11.4 (1H, -NH),  8.00- 6.8 H), 6.2 (1H, -CH),  3.05 (1H, -CH₂), 2.0(3H, -CH₃).  2.0 (6H, -OCH₃) ¹³C NMR 127.4, 133.5, 12.4, 147.1, 120.9, 128.8, 164, 161, 19.9, 170.3, 44.5, 95.3, 61, 134, 128.7, 128.0, 128.7, 126.4, 111.6, 128.8, 127.8 MASS (ESI) m/z: 526 [M+H]⁺.

3g: N-(2-methyl-4-oxoquinazolin-3(4H)-yl)-3-oxo-3-(3-phenyl -5-p-tolyl-2H-pyrazol-1(5 H)-yl) propanamide.

IR: 3380 (-NH), 3345 (-NH),  3125 (-CH), 2975(-CH₃), 2855 (-CH),1690 (C=O), 1700 (C=O), 1535 (C=N), 1240 (C-N).¹H NMR: 11.2 (1H, -NH), 10.3 (1H, -NH),  8.0- 6.8 (13H Ar), 6.5 (1H, -CH),  3.6 (1H, -CH₂), 2.45 (3H, -CH₃),  1.7 (3H, -CH₃) ¹³C NMR 127.4, 133.5, 12.4, 147.1, 120.9, 128.8, 164, 161, 19.9, 170.3, 44.5, 95.3, 61, 134, 128.7, 128.0, 128.7, 126.4, 24.3, 126.4, 128.7 MASS (ESI) m/z: 478 [M+H]⁺.

3h: N-(2-methyl-4-oxoquinazolin-3(4H)-yl)-3-oxo-3-(3,5-di p-tolyl-2H-pyrazol-1(5 H)-yl) propanamide.

IR: 3385 (-NH), 3340 (-NH),  3105 (-CH), 2955 (-CH₃), 2875 (-CH),1720 (C=O), 1715 (C=O), 1565 (C=N), 1235 (C-N).¹H NMR: 12.3 (1H, -NH), 11.4 (1H, -NH),  8.0- 6.8 (13H, Ar), 6.3 (1H, -CH),  3.0 (1H, -CH₂), 2.05(3H, -CH₃), 1.9 (6H, -CH₃) ¹³C NMR 127.4, 133.5, 12.4, 147.1, 120.9, 128.8, 164, 161, 19.9, 170.3, 44.5, 95.3, 61, 134, 128.7, 128.0, 128.7, 126.4, 126.3, 24.3, 24.3 Mass (ESI) m/z: 494 [M+H]⁺.

3i: 3-(3-(3,4-methoxyphenyl)-5-p-tolyl-2H –pyrazol-1(5 H)-yl-N-(2-methyl-4-oxoquinazolin-3(4H)-yl)-3-oxopropanamide

IR: 3425 (-NH), 3390 (-NH),  3145 (-CH), 2930 (-CH₃), 2880 (-CH),1720 (C=O), 1700 (C=O), 1570 (C=N), 1295 (C-N), 1235 (C-O-C-OCH₃).¹H NMR: 12.0 (1H, -NH), 11.1 (1H, -NH),  8.0- 6.7 (13H, Ar), 6.0 (1H, -CH),  3.0 (1H, -CH₂), 2.40 (3H, -CH₃), 2.0 (3H, -OCH₃), 1.85  (3H, -CH₃) ¹³C NMR 127.4, 133.5, 12.4, 147.1, 120.9, 128.8, 164, 161, 19.9, 170.3, 44.5, 95.3, 61, 134, 128.7, 128.0, 128.7, 126.4, 127.4, 24.3, 55.9 Mass (ESI) m/z: 510 [M+H]⁺.

3j: N-(2-methyl-4-oxoquinazolin-3(4H)-yl)-3-(3-(4-nitrophenyl)-5- p-tolyl-2H-pyrazol-1(5 H)-yl) -3-oxopropanamide

IR: 3370 (-NH), 3337(-NH),  3140 (-CH), 2920 (-CH₃), 2885 (-CH),1710 (C=O Str), 1700 (C=O), 1580(-NO₂), 1555 (C=N), 1280 (C-N).¹H NMR: 12.0 (1H, -NH), 11.2 (1H, -NH),  8.1- 6.8(13H, Ar), 6.5 (1H, -CH),  3.5 (1H, -CH₂), 2.5 (3H, -CH₃), 1.5 (3H, -CH₃) ¹³C NMR 127.4, 133.5, 12.4, 147.1, 120.9, 128.8, 164, 161, 19.9, 170.3, 44.5, 95.3, 61, 134, 128.7, 128.0, 128.7, 126.4, 121, 127.3, 24.3 MASS (ESI) m/z: 530 [M+H]⁺.

3k: : 3-(3-(4-chlorophenyl)-5-p-tolyl-2H –pyrazol-1(5 H)-yl-N-(2-methyl-4-oxoquinazolin-3(4H)-yl)-3-oxopropanamide

IR: 3375 (-NH), 3330 (-NH),  3110 (-CH), 2935 (-CH₃), 2836 (-CH), 1700  (C=O), 1690 (C=O), 1555 (C=N), 1290 (C-N), 878 (C-Cl).¹H NMR 6.3 (1H, -CH),  3.1 (1H, -CH₂), 2.5 (3H, -CH₃), 1.9 (3H, -CH₃) ¹³C NMR 127.4, 133.5, 12.4, 147.1, 120.9, 128.8, 164, 161, 19.9, 170.3, 44.5, 95.3, 61, 134, 128.7, 128.0, 128.7, 126.4, 128.8, 127.8, 24.3 MASS (ESI) m/z: 514 [M+H]⁺.

3l: 3-(3-(3,4-dimethoxyphenyl)-5-p-tolyl-2H –pyrazol-1(5 H)-yl-N-(2-methyl-4-oxoquinazolin-3(4H)-yl)-3-oxopropanamide

IR: 3385(NH) 3360 (NH) 3115 (CH) 2925 (CH₃) 2889 (CH) 1700 (C=O), 1690 (C=O), 1555 (C=N), 1290 (C-N), 1225 (OCH₃).¹H-NMR: 12.2 (1H, -NH), 11.3 (1H, -NH),  8.2- 6.8 9(13H, Ar), 6.15(1H, -CH) 3.3 (1H, -CH₂),2.5 (6H, -OCH₃), 2.7 (3H, -CH₃), 1.8 (3H, -CH₃) ¹³C NMR 127.4, 133.5, 12.4, 147.1, 120.9, 128.8, 164, 161, 19.9, 170.3, 44.5, 95.3, 61, 134, 128.7, 128.0, 128.7, 126.4, 24.3, 56.2, 56.2 MASS (ESI) m/z: 540 [M+H]⁺.

Results and Discussion

Chemistry

The research is focused on the precise synthesis of novel N-(2-methyl-4-oxoquinazolin-3(4H)-yl)-3-oxo-3-(3,5-diphenyl-2H-pyrazol-1(5 H)-yl) propanamide (3a-l). The step1 is synthesized chalcones by using Claisen–Schmidt condensation and step2 are synthesized Quinazoline malonic dihydrazide step3 undergoes cyclization Schiff and Mannish base mechanism formation of pyrazole and its scaffold (3a-l) and are confirmed by using different spectral analysis tools ¹HNMR, ¹³CNMR, IR and Mass. The completed scheme is represented in Figure 3 and the Physical properties are shown in Table 1

Figure 3: Scheme I of novel pyrazole linked with quinazoline scaffolds (3a-3l). Click here to View Figure

Table 1: Physical properties of novel pyrazole linked with quinazoline scaffolds (3a-3l).

Compounds	Molecular Formula	Molecular Weight	R	R1	R2	Melting Point	Yield %
3a	C27H23N5O3	465.15	H	H	H	2460C	78%
3b	C28H25N5O3	479.12	CH3	H	H	2870C	88%
3c	C28H25N5O4	495.08	OCH3	H	H	3240C	86%
3d	C27H22N6O5	510.07	NO2	H	H	2760C	78%
3e	C27H22N6O3Cl	499.9	Cl	H	H	2450C	84%
3f	C29H27N5O5	525.16	OCH3	OCH3	H	3450C	72%
3g	C28H25N5O3	479.11	H	H	CH3	2260C	85%
3h	C29H27N5O3	493.02	CH3	H	CH3	2450C	78%
3i	C29H27N5O4	509.02	OCH3	H	CH3	2800C	90%
3j	C28H24N6O5	529.01	NO2	H	CH3	2520C	89%
3k	C28H24N5O3Cl	513.18	Cl	H	CH3	2450C	78%
3l	C30H29N5O5	539.13	OCH3	OCH3	CH3	3150C	82%

 

Biological Activity

In-vitro Anticancer Activity

In our research. A Novel series of compounds are synthesized and evaluated as in-vitro anticancer activity (3a-l) were assessed against three cell lines (MCF7) and (PC-3) and (HT-29) by using MTT assay method 3d and 3e were found potent against all the three cell lines  IC₅₀(µM) 3d (16.52,13.24.10.15 μg/ml) 3e (17.28, 15.26, 12.33 μg/ml) compared with standard drug Doxorubicin (15.29, 12.26, 9.06 μg/ml) and 5-Fluorouracil (16.15, 13.73, 10.25 μg/ml). The IC₅₀ values are represented in Table 2 and the standard graph is represented in Figure 4

Table 2: Anticancer activity of novel pyrazole linked with quinazoline scaffolds IC₅₀ values (3a-3l).

COMPOUNDS	IC50 (µM)
	MCF-7	PC-3	HT-29
3a	39.26 ± 0.32	34.32 ± 0.41	31.26 ± 0.49
3b	45.59 ± 0.51	62.18 ± 0.25	58.15 ± 0.16
3c	95.23 ± 0.86	94.16 ± 0.48	92.23 ± 0.27
3d	16.52 ± 0.052	13.24 ± 0.044	10.15 ± 0.032
3e	17.28 ± 0.065	15.26 ± 0.072	12.33 ± 0.068
3f	85.19 ± 0.16	90.69 ± 0.58	82.22 ± 0.32
3g	46.02 ± 0.27	45.34 ± 0.32	43.26 ± 0.62
3h	88.21 ± 0.48	85.26 ± 0.51	83.45 ± 0.38
3i	35.12 ± 0.21	36.44 ± 0.64	33.19 ± 0.55
3j	48.14 ± 0.71	42.14 ± 0.16	44.28 ± 0.67
3k	47.25 ± 0.25	42.19 ± 0.56	39.15 ± 0.42
3l	92.45 ± 0.68	87.21 ± 0.69	83.24 ± 0.22
DOXORUBICIN	15.29 ± 0.032	12.26 ± 0.041	9.06 ± 0.028
5-FLUROURACIL	16.15 ± 0.016	13.73 ± 0.025	10.25 ± 0.018

 

Figure 4: Graphical representation of Anticancer activity of novel pyrazole linked with quinazoline scaffolds Click here to View Figure

In-vitro Antibacterial activity

   In our research in vitro Antibacterial activity was screened. The novel series of compounds are synthesized (3a-l) was examined against two gram-positive and two gram-negative bacteria strains by using the agar plate method. Interestingly, (3d, 3e,3j,3k) were potent and active against both the bacterial strains. The percentage zone of inhibition values are represented in Table 3 The results show that the substituted of 3d Nitrogen, 3e Chlorine, 3j Methyl and Nitrogen, 3k Methyl and Chlorine substituted compound shows a promising activity comparison with ciprofloxacin.  

Table 3: Antibacterial activty of novel pyrazole linked with quinazoline scaffolds (3a-3l).

	Gram-positive bacteria				Gram-negative bacteria
	Staphylococcus Aureus		Bacillus Subtilis		Escherichia Coli		Pseudomonas Aeruginosa
Compounds	50µg/ml	100µg/ml	50µg/ml	100µg/ml	50g/ml	100µg/ml	50µg/ml	100µg/ml
3a	20	23	8	19	18	21	20	14
3b	14	17	11	13	10	12	12	16
3c	11	17	9	12	13	14	15	18
3d	25	30	24	27	24	28	27	30
3e	25	31	23	27	25	29	27	31
3f	7	10	10	13	15	18	17	20
3g	12	15	15	17	10	12	14	19
3h	11	14	10	12	15	11	8	18
3i	9	10	12	8	13	8	12	10
3j	22	27	18	20	21	22	19	25
3k	23	29	19	21	23	26	22	27
3l	14	15	10	13	9	12	11	14
Ciprofloxacin	28	33	25	29	27	32	30	34

 

Insilico Molecular docking

The insilico molecular docking was analyzed by using Autodock vina PyRx (0.8).The Autodock vina tool shows the accurate result, whenever software is run for trail the docking and binding affinity values are standard and accurate. The protein was downloaded from WWW.RCSBPDB.COM The protein and ligands are uploaded in PyRx tool and adjust coordinates of the x and y-axis. Then, This tool detects interaction with different amino acids and binding affinity values have been predicted in Excel file. Finally obtained binding affinity values and 2D and 3D images. It can be viewed by using tools like Discovery Studio and Molegro Molecular Viewer. The binding affinity values are represented in Table 4 and interaction with different amino acids is represented in figure 5 and 6

Table 4: Insilco molecular docking binding affinity values of novel pyrazole linked with quinazoline scaffolds (3a-3l).

BINDING -AFFINITY Kcal/mol
COMPOUNDS	5C5S	6XXN	3K46
3a	-10.3	-10.3	-11.2
3b	-10.9	-10.7	-10.2
3c	-10.5	-10.2	-10.5
3d	-13.5	-12.8	-13.3
3e	-12.8	-11.9	-12.6
3f	-9.5	-10.8	-10.6
3g	-10.9	-10.6	-11.4
3h	-11.3	-10.8	-11.7
3i	-10.9	-10.4	-10.7
3j	-11.2	-10.8	-12.2
3k	-11.1	-11.0	-11.8
3l	-11.0	-10.1	-10

 

In our research. Insilico Molecular docking was performed and obtained good binding affinity values against PDB ID 5C5S (human myosin 9b RhoGAP), 6XXN (sting CTD complex), 3K46 (E.Coli beta glucurinadase). The binding affinity of the ligand 3d and 3e against proteins is around -12 kcal/mol. The ligand coupled with various amino acids Thr 112(A), Gln 108(A), Asn 139(A), Tyr 111(A), Gly 110(A), Ala 391(A) and Ile 143(A). The molecules binds to protein through its 2D and 3D complex. The protein and ligand bind and coupled with different amino acid interactions are completely moulded each other. In figure the ligand and protein mould through its 3D complex.

Figure 5: Interaction image of 3d with human myosin 9b RhoGAP (Pdb id – 5C5S) Click here to View Figure

Figure 6: Interaction image of 3e with E.Coli beta glucurinadase (Pdb id – 3k46). Click here to View Figure

Conclusion

In this research, The Novel series of compounds are synthesized N-(2-methyl-4-oxoquinazolin-3(4H)-yl)-3-oxo-3-(3,5-diphenyl-2H-pyrazol-1(5H)-yl) propanamide (3a-l). The synthesized derivatives are characterized by ¹HNMR, ¹³CNMR, IR, and MASS. It was evaluated as invitro anticancer and antibacterial activity. Among the series of compounds 3d and 3e exhibit potent against three cancer cell lines MCF7, PC-3, HT-29. IC₅₀(µM) 3d (16.52, 13.24, 10.15 μg/ml) 3e (17.28, 15.26, 12.33 μg/ml) with standard drugs doxorubicin (15.29, 12.26, 9.06 μg/ml) and 5-fluorouracil (16.15, 13.73, 10.25 μg/ml). And also screened antibacterial activity by using agar plate diffusion methods 3d, 3e, 3j, 3k scaffolds exhibit a promising activity compared with the standard drug ciprofloxacin. In our research interestingly withdrawing group attached with scaffolds it shows a more active anti-cancer activity as well as antibacterial activity and also shows based on withdrawing capability. Insilico Molecular Docking performed and it is predicted a good binding affinity against 5C5S(Human-myosin9b-Rhogap), 6XXN (sting CTD complex), 3K46 (E.Coli beta glucurinadase) proteins.

Acknowledgement

The authors are grateful to the director of GITAM for Providing a Lab facility and IICT for providing spectral analysis.

Conflicts of Interest

There are no conflicts of interest declared by the authors.

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