Synthesis and biological evaluation of novel (E)-N'-benzylidene hydrazides as novel c-Met inhibitors through fragment based virtual screening.
ABSTRACT: C-Met plays a crucial role in the development and progression of neoplastic disease. Type II c-Met inhibitors recognise the inactive DFG-out conformation of the kinase, result in better anti-tumour effects due to synergistic effect against the other kinases. According to our previous works, an (E)-N'-benzylidene group was selected as the initial fragment. Two series of (E)-N'-benzylidene hydrazides were designed by fragment growth method. The inhibitory activities were in vitro investigated against c-Met and VEGFR-2. Compound 10b exhibited the most potent inhibitory activity against the c-Met inhibitor (IC50 = 0.37 nM). Compound 11b exhibited multi-target c-Met kinase inhibitory activity as a potential type II c-Met inhibitor (IC50 = 3.41 nM against c-Met; 25.34 nM against VEGFR-2). The two compounds also demonstrate the feasibility of fragment-based virtual screening method for drug discovery.
Project description:Vascular endothelial growth factor receptor-2 (VEGFR-2) plays a critical role in cancer angiogenesis. Inhibition of VEGFR-2 activity proved effective suppression of tumour propagation. Accordingly, two series of new 3-methylquinoxaline derivatives have been designed and synthesised as VEGFR-2 inhibitors. The synthesised derivatives were evaluated <i>in vitro</i> for their cytotoxic activities against MCF-7and HepG2 cell lines. In addition, the VEGFR-2 inhibitory activities of the target compounds were estimated to indicate the potential mechanism of their cytotoxicity. To a great extent, the results of VEGFR-2 inhibition were highly correlated with that of cytotoxicity. Compound <b>27a</b> was the most potent VEGFR-2 inhibitor with IC<sub>50</sub> of 3.2 nM very close to positive control sorafenib (IC<sub>50</sub> = 3.12 nM). Such compound exhibited a strong cytotoxic effect against MCF-7 and HepG2, respectively with IC<sub>50</sub> of 7.7 and 4.5 µM in comparison to sorafenib (IC<sub>50</sub> = 3.51 and 2.17 µM). In addition, compounds <b>28</b>, <b>30f</b>, <b>30i</b>, and <b>31b</b> exhibited excellent VEGFR-2 inhibition activities (IC<sub>50</sub> range from 4.2 to 6.1 nM) with promising cytotoxic activity. Cell cycle progression and apoptosis induction were investigated for the most active member <b>27a</b>. Also, the effect of <b>27a</b> on the level of caspase-3, caspase-9, and BAX/Bcl-2 ratio was determined. Molecular docking studies were implemented to interpret the binding mode of the target compounds with the VEGFR-2 pocket. Furthermore, toxicity and ADMET calculations were performed for the synthesised compounds to study their pharmacokinetic profiles.
Project description:In this study, a set of novel benzoxazole derivatives were designed, synthesised, and biologically evaluated as potential VEGFR-2 inhibitors. Five compounds (<b>12d</b>, <b>12f</b>, <b>12i</b>, <b>12l</b>, and <b>13a</b>) displayed high growth inhibitory activities against HepG2 and MCF-7 cell lines and were further investigated for their VEGFR-2 inhibitory activities. The most potent anti-proliferative member <b>12 l (</b>IC<sub>50</sub> = 10.50 μM and 15.21 μM against HepG2 and MCF-7, respectively<b>)</b> had the most promising VEGFR-2 inhibitory activity (IC<sub>50</sub> = 97.38 nM). A further biological evaluation revealed that compound <b>12l</b> could arrest the HepG2 cell growth mainly at the Pre-G1 and G1 phases. Furthermore, compound <b>12l</b> could induce apoptosis in HepG2 cells by 35.13%. likely, compound <b>12l</b> exhibited a significant elevation in caspase-3 level (2.98-fold) and BAX (3.40-fold), and a significant reduction in Bcl-2 level (2.12-fold). Finally, docking studies indicated that <b>12l</b> exhibited interactions with the key amino acids in a similar way to sorafenib.
Project description:Based on quinazoline, quinoxaline, and nitrobenzene scaffolds and on pharmacophoric features of VEGFR-2 inhibitors, 17 novel compounds were designed and synthesised. VEGFR-2 IC<sub>50</sub> values ranged from 60.00 to 123.85 nM for the new derivatives compared to 54.00 nM for sorafenib. Compounds <b>15<sub>a</sub></b>, <b>15<sub>b</sub></b>, and <b>15<sub>d</sub></b> showed IC<sub>50</sub> from 17.39 to 47.10 µM against human cancer cell lines; hepatocellular carcinoma (HepG2), prostate cancer (PC3), and breast cancer (MCF-7). Meanwhile, the first in terms of VEGFR-2 inhibition was compound <b>15<sub>d</sub></b> which came second with regard to antitumor assay with IC<sub>50</sub> = 24.10, 40.90, and 33.40 µM against aforementioned cell lines, respectively. Furthermore, Compound <b>15<sub>d</sub></b> increased apoptosis rate of HepG2 from 1.20 to 12.46% as it significantly increased levels of Caspase-3, BAX, and P53 from 49.6274, 40.62, and 42.84 to 561.427, 395.04, and 415.027 pg/mL, respectively. Moreover, <b>15<sub>d</sub></b> showed IC<sub>50</sub> of 253 and 381 nM against HER2 and FGFR, respectively.
Project description:Herein, a new wave of bis([1, 2, 4]triazolo)[4,3-<i>a</i>:3',4'-<i>c</i>]quinoxaline derivatives have been successfully designed and synthesised. The synthesised derivatives were biologically investigated for their cytotoxic activities against HepG2 and MCF-7. Also, the tested compounds were further examined <i>in vitro</i> for their VEGFR-2 inhibitory activity. The most promising derivative <b>23j</b> was further investigated for its apoptotic behaviour in HepG2 cell lines using flow cytometric and western-plot analyses. Additional <i>in-silico</i> studies were performed to predict how the synthesised compounds can bind to VEGFR-2 and to determine the drug-likeness profiling of these derivatives. The results revealed that compounds <b>23a</b>, <b>23i</b>, <b>23j</b>, <b>23l</b>, and <b>23n</b> displayed the highest antiproliferative activities against the two cell lines with IC<sub>50</sub> values ranging from 6.4 to 19.4 µM. Furthermore, compounds <b>23a</b>, <b>23d</b>, <b>23h</b>, <b>23i</b>, <b>23j</b>, <b>23l</b>, <b>23 m</b>, and <b>23n</b> showed the highest VEGFR-2 inhibitory activities with IC<sub>50</sub> values ranging from 3.7 to 11.8 nM, comparing to sorafenib (IC<sub>50</sub> = 3.12 nM). Moreover, compound <b>23j</b> arrested the HepG2 cell growth at the G2/M phase and induced apoptosis by 40.12% compared to the control cells (7.07%). As well, such compound showed a significant increase in the level of caspase-3 (1.36-fold), caspase-9 (2.80-fold), and BAX (1.65-fold), and exhibited a significant decrease in Bcl-2 level (2.63-fold).
Project description:Fibroblast growth-factor receptor (FGFR) is a potential target for cancer therapy. We designed three novel series of FGFR1 inhibitors bearing indazole, benzothiazole, and 1<i>H</i>-1,2,4-triazole scaffold <i>via</i> fragment-based virtual screening. All the newly synthesised compounds were evaluated <i>in vitro</i> for their inhibitory activities against FGFR1. Compound <b>9d</b> bearing an indazole scaffold was first identified as a hit compound, with excellent kinase inhibitory activity (IC<sub>50</sub> = 15.0 nM) and modest anti-proliferative activity (IC<sub>50</sub> = 785.8 nM). Through two rounds of optimisation, the indazole derivative <b>9 u</b> stood out as the most potent FGFR1 inhibitors with the best enzyme inhibitory activity (IC<sub>50</sub> = 3.3 nM) and cellular activity (IC<sub>50</sub> = 468.2 nM). Moreover, <b>9 u</b> also exhibited good kinase selectivity. In addition, molecular docking study was performed to investigate the binding mode between target compounds and FGFR1.
Project description:VEGFR-2, the subtype receptor tyrosine kinase (RTK) responsible for angiogenesis, is expressed in various cancer cells. Thus, VEGFER-2 inhibition is an efficient approach for the discovery of new anticancer agents. Accordingly, a new set of nicotinamide derivatives were designed and synthesized to be VEGFR-2 inhibitors. The chemical structures were confirmed using IR, <sup>1</sup>H-NMR, and <sup>13</sup>C-NMR spectroscopy. The obtained compounds were examined for their anti-proliferative activities against the human cancer cell lines (HCT-116 and HepG2). VEGFR-2 inhibitory activities were determined for the titled compounds. Compound <b>8</b> exhibited the strongest anti-proliferative activities with IC<sub>50</sub> values of 5.4 and 7.1 µM against HCT-116 and HepG2, respectively. Interestingly, compound <b>8</b> was the most potent VEGFR-2 inhibitor with an IC<sub>50</sub> value of 77.02 nM (compare to sorafenib: IC<sub>50</sub> = 53.65 nM). Treatment of HCT-116 cells with compound <b>8</b> produced arrest of the cell cycle at the G0-G1 phase and a total apoptosis increase from 3.05 to 19.82%-6.5-fold in comparison to the negative control. In addition, compound <b>8</b> caused significant increases in the expression levels of caspase-8 (9.4-fold) and Bax (9.2-fold), and a significant decrease in the Bcl-2 expression level (3-fold). The effects of compound <b>8</b> on the levels of the immunomodulatory proteins (TNF-α and IL-6) were examined. There was a marked decrease in the level of TNF-α (92.37%) compared to the control (82.47%) and a non-significant reduction in the level of IL-6. In silico docking, molecular dynamics simulations, and MM-PBSA studies revealed the high affinity, the correct binding, and the optimum dynamics of compound 8 inside the active site of VEGFR-2. Finally, in silico ADMET and toxicity studies indicated acceptable values of drug-likeness. In conclusion, compound <b>8</b> has emerged as a promising anti-proliferative agent targeting VEGFR-2 with significant apoptotic and immunomodulatory effects.
Project description:The role of metalloenzymes in tumor progression had broadened their application in cancer therapy. Of these, MMPs and CAs are validated druggable targets that share some pivotal signaling pathways. The majority of MMPs or CAs inhibitors are designed as single-target agents. Despite their transient efficacy, these agents are often susceptible to resistance. This set the stage to introduce dual inhibitors of correlated MMPs and CAs. The next step is expected to target the common vital signaling nodes as well. In this regard, VEGFR-2 is central to various tumorigenesis events involving both families, especially MMP-2 and CA II. Herein, we report simultaneous inhibition of MMP-2, CA II, and VEGFR-2 via rationally designed hybrid 1,2,4-triazolo[4,3-<i>a</i>]pyrimidinone acyclo C-nucleosides. The promising derivatives were nanomolar inhibitors of VEGFR-2 (<b>8</b>; IC<sub>50</sub> = 5.89 nM, <b>9</b>; IC<sub>50</sub> = 10.52 nM) and MMP-2 (<b>8</b>; IC<sub>50</sub> = 17.44 nM, <b>9</b>; IC<sub>50</sub> = 30.93 nM) and submicromolar inhibitors of CA II (<b>8</b>; IC<sub>50</sub> = 0.21 µM, <b>9</b>; IC<sub>50</sub> = 0.36 µM). Docking studies predicted their binding modes into the enzyme active sites and the structural determinants of activity regarding substitution and regioselectivity. MTT assay demonstrated that both compounds were 12 folds safer than doxorubicin with superior anticancer activities against three human cancers recording single-digit nanomolar IC<sub>50</sub>, thus echoing their enzymatic activities. Up to our knowledge, this study introduces the first in class triazolopyrimidinone acyclo C-nucleosides VEGFR-2/MMP-2/CA II inhibitors that deserve further investigation.
Project description:There is an urgent need to design new anticancer agents that can prevent cancer cell proliferation even with minimal side effects. Accordingly, two new series of 3-methylquinoxalin-2(1<i>H</i>)-one and 3-methylquinoxaline-2-thiol derivatives were designed to act as VEGFR-2 inhibitors. The designed derivatives were synthesised and evaluated <i>in vitro</i> as cytotoxic agents against two human cancer cell lines namely, HepG-2 and MCF-7. Also, the synthesised derivatives were assessed for their VEGFR-2inhibitory effect. The most promising member <b>11e</b> were further investigated to reach a valuable insight about its apoptotic effect through cell cycle and apoptosis analyses. Moreover, deep investigations were carried out for compound <b>11e</b> using western-plot analyses to detect its effect against some apoptotic and apoptotic parameters including caspase-9, caspase-3, BAX, and Bcl-2. Many <i>in silico</i> investigations including docking, ADMET, toxicity studies were performed to predict binding affinity, pharmacokinetic, drug likeness, and toxicity of the synthesised compounds. The results revealed that compounds <b>11e, 11g, 12e, 12g,</b> and <b>12k</b> exhibited promising cytotoxic activities (IC<sub>50</sub> range is 2.1 - 9.8 µM), comparing to sorafenib (IC<sub>50</sub> = 3.4 and 2.2 µM against MCF-7 and HepG2, respectively). Moreover, <b>11b, 11f, 11g, 12e, 12f, 12g,</b> and <b>12k</b> showed the highest VEGFR-2 inhibitory activities (IC<sub>50</sub> range is 2.9 - 5.4 µM), comparing to sorafenib (IC<sub>50</sub> = 3.07 nM). Additionally, compound <b>11e</b> had good potential to arrest the HepG2 cell growth at G2/M phase and to induce apoptosis by 49.14% compared to the control cells (9.71%). As well, such compound showed a significant increase in the level of caspase-3 (2.34-fold), caspase-9 (2.34-fold), and BAX (3.14-fold), and a significant decrease in Bcl-2 level (3.13-fold). For <i>in silico</i> studies, the synthesised compounds showed binding mode similar to that of the reference compound (sorafenib).
Project description:A series of benzylidene analogs of oleanolic acid <b>4a∼4s</b> were synthesized and assessed for their <i>α</i>-glucosidase and <i>α</i>-amylase inhibitory activities. The results presented that all synthesized analogs exhibited excellent-to-moderate inhibitory effects on <i>α</i>-glucosidase and <i>α</i>-amylase. Analog <b>4i</b> showed the highest <i>α</i>-glucosidase inhibition (IC<sub>50</sub>: 0.40 μM), and analog <b>4o</b> presented the strongest <i>α</i>-amylase inhibition (IC<sub>50</sub>: 9.59 μM). Inhibition kinetics results showed that analogs <b>4i</b> and <b>4o</b> were reversible and mixed-type inhibitors against <i>α</i>-glucosidase and <i>α</i>-amylase, respectively. Simulation docking results demonstrated the interaction between analogs and two enzymes. Moreover, analogs <b>4i</b> and <b>4o</b> showed a high level of safety against 3T3-L1 and HepG2 cells.
Project description:Thirteen (<i>Z</i>)-2-(substituted benzylidene)benzimidazothiazolone analogs were synthesized and evaluated for their inhibitory activity against mushroom tyrosinase. Among the compounds synthesized, compounds <b>1</b>-<b>3</b> showed greater inhibitory activity than kojic acid (IC<sub>50</sub> = 18.27 ± 0.89 μM); IC<sub>50</sub> = 3.70 ± 0.51 μM for <b>1</b>; IC<sub>50</sub> = 3.05 ± 0.95 μM for <b>2</b>; and IC<sub>50</sub> = 5.00 ± 0.38 μM for <b>3</b>, and found to be competitive tyrosinase inhibitors. In silico molecular docking simulations demonstrated that compounds <b>1</b>-<b>3</b> could bind to the catalytic sites of tyrosinase. Compounds <b>1</b>-<b>3</b> inhibited melanin production and cellular tyrosinase activity in a concentration-dependent manner. Notably, compound <b>2</b> dose-dependently scavenged ROS in B16F10 cells. Furthermore, compound <b>2</b> downregulated the protein kinase A (PKA)/cAMP response element-binding protein (CREB) and mitogen-activated protein kinase (MAPK) signaling pathways, which led to a reduction in microphthalmia-associated transcription factor (MITF) expression, and decreased tyrosinase, tyrosinase related protein 1 (TRP1), and TRP2 expression, resulting in anti-melanogenesis activity. Hence, compound <b>2</b> may serve as an anti-melanogenic agent against hyperpigmentation diseases.