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. 2024 May 22;19(5):e0303186. doi: 10.1371/journal.pone.0303186

3,3’-((3,4,5-trifluoropHenyl)methylene)bis(4-hydroxy-2H-chromen-2-one) inhibit lung cancer cell proliferation and migration

Wenhui Luo 1,2, Guoxin Chang 3, Dingmei Lin 3, Hongyi Xie 3, Huilong Sun 1, Zhibin Li 1, Shirong Mo 1, Ruixue Wang 1,*, Yan Wang 3,*, Zhaoguang Zheng 1,*
Editor: Wagdy M Eldehna4
PMCID: PMC11111047  PMID: 38776295

Abstract

Lung cancer is a major public health challenge and, despite therapeutic improvements, is the first leading cause of cancer worldwide. The current cure rate from advanced cancer treatment is excessively low. Therefore, it is of great importance to identify novel, potent and less toxic anticancer agents for the treatment of lung cancer. The aim of our research is to synthesize a new biscoumarin 3,3’-((3,4,5-trifluorop -phenyl)methylene)bis(4-hydroxy-2H-chromen-2-one) (C35) as an anticancer agent. C35 was simply prepared by 4-hydroxycoumarin and 3,4,5-trifluorobenzaldehyde under ethanol and its structure was analyzed by spectroscopic analyses. The anti-proliferation effect of C35 was detected using CCK-8 assay. Migration abilities were measured by Transwell assay. The expression of correlated proteins was determined by Western blot. The results showed that C35 displayed strong cytostatic effects on lung cancer cell proliferation. In addition, C35 possessed a significant inhibition of migration by reducing the expression of matrix metalloproteinases-2 (MMP-2) and MMP-9 in lung cancer cells. Furthermore, C35 treatment suppressed the phosphorylation of p38 in lung cancer cells. Moreover, in vivo experiments were carried out, in which we treated Lewis tumor-bearing C57 mice via intraperitoneal injection of C35. Results showed that C35 inhibited tumor growth in vivo. In conclusion, our study demonstrated the anticancer activity of C35 via suppression of lung cancer cell proliferation and migration, which is possibly involved with the inhibition of the p38 pathway.

Introduction

Lung cancer is the most common cause of cancer death, accounting for 24% of cancer-related deaths, with a 5-year survival rate of only 15% after diagnosis. Treatment options for lung cancer include surgery, chemotherapy, radiotherapy, immunotherapy and targeted therapy. In recent years, significant progress has been made in immunotherapy and targeted therapy for lung cancer, but the 5-year survival rate of lung cancer patients is still low, and lung cancer is prone to recurrence and metastasis. Therefore, it is of great importance to identify novel, potent, and less toxic anticancer agents for lung cancer treatment.

The natural products are important sources of drugs with a significant proportion of current drugs being natural products or derived from natural products. Coumarin, also known as 1,2-benzopyranone, has a large conjugated system in which the benzene ring and the pyranone ring in its structure can form a large conjugated system, making coumarin highly modifiable and capable of introducing a variety of functional groups [1]. Coumarin-derived compounds obtained through total synthesis or structural modification can enhance their anti-tumor activity. Accumulating data have shown a number of synthesized coumarin-derived compounds for their potential anti-tumor activities [1]. Those coumarins-based anticancer agents have been identified for a variety of mechanisms of action, including alkylating agents, topoisomerase inhibitors, angiogenesis inhibitors, apoptosis inducers, human carbonic anhydrase inhibitors, telomerase inhibitors and miscellaneous agent [1].

Biscoumarin, a coumarin-derived compound, is mainly used as an anticoagulant for the prevention and treatment of thrombosis. Recently the anti-diabetic [2] and anti-tumor [3] effects of biscoumarin have also been reported. Methylenebis (4-hydroxy-2H-chromen-2-one) biscoumarins are easily and simply synthesized by 4-hydroxycoumarin and different substituent benzaldehydes. Different substituent benzaldehydes affect the efficiency of synthesis and the activity of the product. Therefore, the selection of benzaldehyde with different substituents is particularly important. Previously, we found that the synthesized biscoumarin 3,3’-((4-chlorophenyl)methylene)bis(4-hydroxy-2H-chromen-2-one) suppressed non-small cell lung cancer cell proliferation and induced cell apoptosis, possibly involving receptor interacting protein-1 [4]. Fluorine and chlorine are both halogen elements, and the rational design by the introduction of fluorine into a compound has achieved success in the development of organic anticancer drugs [5–7]. So here we synthesized another similar biscoumarin 3,3’-((3,4,5-trifluoropHenyl)methylene)bis(4-hydroxy-2H-chromen-2-one) (compound C35), with three fluorine substituents instead of one chlorine substituent on the benzaldehyde unit, in order to explore more potential candidates for lung cancer treatment.

Materials and methods

Chemicals and apparatus

The established H1299 and Lewis cell lines were obtained from The Chinese Center for Type Culture Collection. NMR data were collected on a Bruker AM-400 spectrometer in DMSO-d6 (Bruker, Fällanden, Switzerland). HR-ESI-MS were performed in MeOH on a thermofisher Q-Fleet spectrometer (Thermofisher Scientific, San Jose, CA, USA). The melting point was detected by RY-2 melting point meter (Tianjin analytical instrument factory, Tianjin, China).

Synthesis of compound biscoumarin (C35)

The synthesis of C35 is according to our previous paper [4]. In detail, 1.62 g (10 mmol) 4-hydroxycoumarin (1) and 5 mmol 3,4,5-trifluorobenzaldehyde (2) were dissolved in 20 mL ethanol, placed in a 50 mL round-bottom flask, stirred and heated at reflux for 4 h, solid precipitation could be seen in the reaction, monitored by thin layer chromatography (TLC) until the end of the reaction, cooled and filtered, the precipitate was recrystallized with ethanol to obtain 3 (C35, Fig 1).

Fig 1. Synthesis of C35.

Fig 1

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Cell culture

H1299, Lewis, H9C2 and Beas-2B cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM; Gibco, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Gibco, Waltham, MA, USA). All cells were cultured at 37°C in a humidified atmosphere containing 5% CO2. The cell authentication was performed via STR profiling and species authentication. All the cells were passaged fewer than 20 times.

Cell proliferation assay

The H1299, Lewis, H2C9 and Beas-2B cells were plated (2 × 103/well) in 96-well plates and incubated with C35 (0, 10, 20 μM) for 72 h, respectively. CCK-8 solutions (Dojindo, Japan) were then added and maintained at 37°C for 1 h. Finally, the absorbance was measured at 450 nm.

Western blot

Western blot was performed according to our previous paper [4]. In brief, whole cell extracts were prepared by lysing the cells in lysis buffer (KeyGEN biotech, Nanjing, China). Then the equal amounts of total proteins were resolved by SDS-PAGE and the proteins of interest were probed by Western blot. Subsequently the Western blot results were visualized by enhanced chemiluminescence according to manufacturer’s instructions (Millipore, Billerica, MA, USA). The expression of protein was quantified by Image J software.

Transwell assay

A Transwell chamber (Corning, MA, United States) was used to analyze cell migration capacity. Briefly, the lung cancer cells in serum-free medium were loaded into the upper chamber and the lower chamber was loaded with medium containing 5% FBS. Then the cells were treated with C35 (0, 10, 20 μM). After 24 h of incubation, non-migration cells were removed and migration cells were stained with crystal violet. Finally, the number of migration cells was counted by microscopy.

ELISA

The amount of MMP-2 and MMP-9 in cell culture supernatants were determined by ELISA according to the manufacturer’s instructions (Zikerbio, Shenzen, China). Briefly, cells were plated on 6-well plates at 70–80% confluence. After overnight culture, cells were treated with C35 (0, 10, 20 μM) for 72 h, then cell culture supernatants were collected and incubated with HRP-labelled detection antibody in 96-well plates. After washing 5 times with washing buffer, 50 μL of substrate A and B were added to each well, followed by 50 μL of stopping solution. OD values were measured at 450 nm.

In vivo study

Six-week-old male mice were purchased from Animal Center (Guangzhou, China) and maintained under pathogen-free conditions. All procedures involving animals and their care were conducted in in accordance with the guidelines of the Institutional Animal Care and Use Committee of Foshan University. A total of 1× 106 cells in 100uL Phosphate Buffered Saline (PBS) was subcutaneously injected in the right flank of the mice. After palpable tumors had developed, mice were randomly divided into three groups and received four intraperitoneal injections of following agents: (a) PBS control; (b) 50 mg/Kg C35; (c) 75 mg/Kg C35. There are 5 mice in each group. The tumor volumes were measured with a caliper and calculated as the following formula: V = 0.5 × length × width2, the length was the long axis of the tumor, and the width was the short axis. At the end of the experiments, mice were euthanized and the excised tumors were collected and weighed.

Statistical analysis

Data are presented as mean ± SD. Statistical analyses were performed with GraphPad PRISM 6.0 software. Significant differences between two groups were compared using a Student’s t-test (two-tailed). A one-way ANOVA was used in multiple comparisons.

Results

Identification of C35

3,3’-((3,4,5-trifluoropHenyl)methylene)bis(4-hydroxy-2H-chromen-2-one) (C-35), 1.89g with the yield of 81%, white power, formula: C25H13F3O6, m.p. 250–252°C; ESI-MS, m/z: 467 [M+H]+; IR (KBr): 3070, 2725,2585, 1671 (C = O), 1604, 1528, 1434, 1349, 1101 cm-1; 1H-NMR (DMSO-d6, 400MHz): d 6.275 (s, 1H, H-11), 7.063 (dd, 2H, J = 6.4, 10.4Hz, H-2′′,6′′), 7.289 (td, 2H, J = 1.2, 8.0 Hz, H-6,6′), 7.338 (dd, 2H, H-8,8′), 7.585 (td, 2H, J = 1.6, 7.6 Hz, H-7,7′), 7.894 (dd, 2H, J = 1.6, 8.0Hz, H-5,5′), 10.685 (brs, 2H, 2OH). Compared to the literature [8], the compound is identified as 3,3’-((3,4,5-trifluoropHenyl)methylene)bis(4-hydroxy-2H-chromen-2-one) (Fig 1, S1-S3 Figs in S1 File).

C35 inhibited the proliferation of lung cancer cells

To evaluate the anti-proliferative effect of C35, human lung cancer cell H1299 and mouse lung cancer cell Lewis cells were treated with various concentrations of C35 for 72 h, respectively, and cell proliferation was measured by CCK-8 assay. We found that C35 inhibited the proliferation of H1299 and Lewis cells in a manner with dose-dependent (Fig 2A and 2B). In H1299 and Lewis, C35 had IC50 values of 20.77μM and 20.87μM, respectively. In addition, the human pulmonary epithelial cell line Beas-2B and normal rat cardiomyocytes cell line H9C2 were used to assess the cytotoxic effect of C35. As shown in Fig 2C and 2D, C35 is less toxic to H9C2 and Beas-2B cells than to H1299 and Lewis cells, which suggests that C35 inhibits lung cancer growth in a specific manner.

Fig 2. The proliferation of lung cancer cells was inhibited by C35.

Fig 2

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The H1299 (A), Lewis (B), H9C2 (C) and Beas-2B (D) cells were treated with C35 at the indicated concentrations (0, 10 and 20 μM) for 72 h. And then the cell viability was measured by CCK-8 assay. Data are presented as mean±SD of three independent experiments. **p<0.01.

C35 reduced the migration of lung cancer cells

Cell migration is a key step in the cancer progression. To further investigate the role of C35 on cell metastasis, the cells were subjected to an Transwell migration assay. As shown in Fig 3, there is a clear trend towards a dose-dependent decrease in migration cells after C35 treatment in both H1299 and Lewis cells.

Fig 3. The migration of lung cancer cells was reduced by C35.

Fig 3

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(A) H1299 and Lewis cells were treated with different concentrations of C35 (0, 10, and 20 μM) for 24 h. (B and C) The migration cells were photographed and quantified. Data are presented as mean±SD of three independent experiments. **p<0.01.

As tumor cells migrate, MMPs degrade the basement membrane [9]. Therefore, the protein expression of MMP-2 and MMP-9 in lung cancer cells in response to C35 treatment were evaluated through Western blot and ELISA. The Western blot analysis showed that the high concentration of C35 (20 μM) stimulation decreased the expression of MMP-9 in H1299 cells (Fig 4A). In addition, C35 stimulation decreased the expression of MMP-9 and MMP-2 in Lewis cells (Fig 4B). Moreover, our data showed that C35 treatment significantly inhibited the amount of MMP-2 and MMP-9 in extracellular fractions in both H1299 and Lewis cells (Fig 4C and 4D). Taken together, those results suggest that C35 reduces lung cancer cell migration.

Fig 4. C35 reduced the expression of MMP-2 and MMP-9.

Fig 4

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(A and B) Western blot analyses of the relative expression of MMP-2 and MMP-9 in H1299 and Lewis cells treated with or without C35 as indicated concentrations. β-actin served as loading control. 1, 2 and 3 represent three of the sample in the indicated group. (C and D) The amount of MMP-2 and MMP-9 in H1299 and Lewis cell culture medias were determined by ELISA. Data are presented as mean±SD of three independent experiments. *p<0.05.**p<0.01.

C35 suppressed the phosphorylation of p38 in lung cancer cells

AKT and p38 pathways has been reported to regulate cell proliferation, migration, and invasion in cancer [10, 11]. Consequently, we investigated whether C35 has an impact on these signaling pathways, and found that it decreased p38 phosphorylation (Fig 5), but not AKT phosphorylation (S4 Fig in S1 File).

Fig 5.

Fig 5

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Western blot analyses of the phosphorylation of p38 in H1299 (A) and Lewis (B) cells treated with or without C35 as the indicated concentrations. β-actin served as loading control. The 1, 2 and 3 represent three of the samples in the indicated group. Data are presented as mean±SD of three independent experiments. *p<0.05.**p<0.01.

C35 inhibited the tumor growth in vivo

As shown in Fig 6A–6C, despite the same number of Lewis cells injected, tumor growth in vivo was significantly reduced in either 50 mg/Kg C35 or 75 mg/Kg C35 treatment groups compared to PBS group, as evidenced by a decrease in tumor growth rate and weight of the excised tumor in C35 treated group compared to those in PBS group. Moreover, C35 did not cause a significant impairment in the bodyweight (Fig 6D) or tissue morphology of mice (S5 Fig in S1 File). All mice survived after treatment with C35 or PBS. In conclusion, consistent with the in vitro results, C35 had the antitumor activity in vivo.

Fig 6. C35 inhibited tumor growth on Lewis allograft tumor in C57 mice.

Fig 6

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(A) Images of Lewis tumor of C57 mice treated with PBS or C35 (n = 5 in each group). (B) The tumor volume was measured every 2 days, and the tumor growth curve was drawn. (C) Tumor weights of the three groups were measured. Each dot represents one sample. (D) Comparison of body weight of C57 mice before and after C35 or PBS treatment. *p<0.05. **p<0.01.

Discussion

Many natural and synthetic coumarin-like compounds have been extensively studied by many researchers for their anticancer activity due to their structural, non-toxic and biological properties [1]. These coumarin-based anticancer drugs have been identified through diverse mechanisms of action, such as alkylating agents [12, 13], topoisomerase inhibitors [14, 15], hormone antagonists [16–19], angiogenesis inhibitors [20–22], antimitotic agents [23–26], apoptosis inducers [27–31], human carbonic anhydrase inhibitors [32–35], telomerase inhibitors [36, 37]. Biscoumarin, a coumarin-derived compound, has been reported as a potent and efficient enzyme inhibitor as α-glucosidase inhibitor, α-amylase inhibitor, urease inhibitor, aromatase inhibitor [38]. Some studies have reported the development and biosynthesis of coumarin derivatives, and showed their anti-proliferative effects on tumor cells [3, 39–43], but the mechanism remains largely unknown.

Here, we synthesized biscoumarin C35 and found that C35 exhibited significant cytotoxicity against the lung cancer cells in a concentration dependent manner, but had little effect on normal cells. Moreover, C35 treatment in vivo did not cause parenchymal organ damage in brains, hearts, lungs, livers and kidneys. In addition, migration ability is related to the metastatic potential of cancer cells, which contributes to cancer progression and poor patient outcomes [44, 45]. Extracellular matrix and basement membrane are the main barriers for tumor metastasis [46]. Degradation of stromal collagen by MMP-2 and MMP-9 is the biochemical basis for tumor cell migration and invasion into surrounding tissues [47]. Therefore, the role of MMP-2 and MMP-9 is considered to be one of the key steps in tumor metastasis. Consistently, our results indicated that C35 dose-dependently inhibited the migration of H1299 and Lewis cells, which concomitant with the reduced expression of MMP-2 and MMP-9.

The MAPK signaling pathway has been reported to play an important role in cell proliferation, apoptosis and differentiation [48]. The p38 signaling pathway, one of the MAPK signaling pathways, plays a central role in regulating the expression and activity of MMPs [49, 50], and is integral in carcinogenesis and cancer maintenance [51, 52]. Activation of the p38 signaling pathway has been showed increases the expression of MMP-2 and MMP-9 [53]. In this study, we observed that C35 significantly repressed the phosphorylation of p38, indicating that C35 may exert its cytotoxic effect by inactivating p38 signaling pathway.

Furthermore, in our previous study [4] the synthetic biscoumarin 3,3’-((4-chlorophenyl)methylene)bis(4-hydroxy-2H-chromen-2-one) suppressed non-small cell lung cancer cell proliferation and induced cell apoptosis. Structure-activity relationship (SAR) research revealed that electron-withdrawing groups such as Cl and NO2 on benzaldehyde showed the most profound anti-cancer activity [54]. Mayank and colleagues [8] have developed furtherly that chloro- substituent at 2 or 6, or 4 position on benzaldehyde provided good anticancer activity. The rational design by the introduction of fluorine into a compound has achieved success in the development of organic anticancer drugs [5–7]. In our present study, although the similar biscoumarin C35 with 3,4,5-trifluoro substituents instead of 4-chloro on the benzaldehyde ring have higher IC50 value in Lewis lung cancer cells, biscoumarin C35 inhibited the migration of lung cancer cells. Most importantly, C35 showed anti-cancer capacity in vivo. It is worth for further study.

Conclusions

In conclusion, our study showed that the biscoumarin C35 with 3,4,5-trifluoro substituents instead of 4-chloro on the benzaldehyde ring displayed strong cytostatic effects on lung cell proliferation, and also possessed a significant inhibition of migration by reducing the expression of MMP-2 and MMP-9 in lung cancer cells. Moreover, C35 treatment suppressed the phosphorylation of p38 in lung cancer cells, which may contribute to the anti-cancer activity of C35. Therefore, C35 may be a novel and effective approach for the treatment of lung cancer.

Supporting information

S1 File

(DOCX)

pone.0303186.s001.docx (1.5MB, docx)
S2 File

(XLSX)

pone.0303186.s002.xlsx (18.7KB, xlsx)
S1 Raw images

(PDF)

pone.0303186.s003.pdf (7.1MB, pdf)

Data Availability

All relevant data are within the manuscript and its Supporting Information files.

Funding Statement

This study was supported by grants from the Science and Technology Key Project of COVID‑19 in Foshan city (No. 2020001000206, ZG Zheng, URL: http://fskjj.foshan.gov.cn/), Regional Joint Fund‑Key Project of Guangdong Basic and Applied Basic Research fund (2020B1515120033, ZG Zheng, URL: http://gdstc.gd.gov.cn/), the Provincial Enterprise Joint Fund General Project of Guangdong Basic and Applied Basic Research fund (2022A1515220132, ZG Zheng, URL: http://gdstc.gd.gov.cn/), the Special Fund for Key Fields of Higher Education of Guangdong Province (2023ZDZX2058, ZG Zheng, URL: https://edu.gd.gov.cn/), and the project of the Scientific and Technological Office of Foshan (2320001007430, RX Wang, URL: http://fskjj.foshan.gov.cn/). The funding sources had no role in the study design, data collection, data analysis, interpretation or writing of the report. There was no additional external funding received for this study.

References

  • 1.Al-Warhi T, Sabt A, Elkaeed EB, Eldehna WM. Recent advancements of coumarin-based anticancer agents: An up-to-date review. Bioorg Chem. 2020;103:104163. Epub 2020/09/06. doi: 10.1016/j.bioorg.2020.104163 . [DOI] [PubMed] [Google Scholar]
  • 2.Asgari MS, Mohammadi-Khanaposhtani M, Kiani M, Ranjbar PR, Zabihi E, Pourbagher R, et al. Biscoumarin-1,2,3-triazole hybrids as novel anti-diabetic agents: Design, synthesis, in vitro alpha-glucosidase inhibition, kinetic, and docking studies. Bioorg Chem. 2019;92:103206. Epub 2019/08/25. doi: 10.1016/j.bioorg.2019.103206 . [DOI] [PubMed] [Google Scholar]
  • 3.Zhou HY, Dong FQ, Du XL, Zhou ZK, Huo HR, Wang WH, et al. Antitumor activities of biscoumarin and dihydropyran derivatives. Bioorg Med Chem Lett. 2016;26(16):3876–80. Epub 2016/07/20. doi: 10.1016/j.bmcl.2016.07.023 . [DOI] [PubMed] [Google Scholar]
  • 4.Wang R, Xie H, Wang X, Liu Y, Su Z, Zheng Z. A synthetic biscoumarin suppresses lung cancer cell proliferation and induces cell apoptosis by increasing expression of RIP1. Chin J Physiol. 2022;65(3):136–42. Epub 2022/07/02. doi: 10.4103/cjp.cjp_107_21 . [DOI] [PubMed] [Google Scholar]
  • 5.Yang Y, Guo L, Ge X, Zhu T, Chen W, Zhou H, et al. The Fluorine Effect in Zwitterionic Half-Sandwich Iridium(III) Anticancer Complexes. Inorganic Chemistry. 2019;59. doi: 10.1021/acs.inorgchem.9b03006 . [DOI] [PubMed] [Google Scholar]
  • 6.Lim YH, Oo CW, Koh RY, Voon GL, Loh YCJDDR. Synthesis, characterization, and anti-cancer activity of new chalcone derivatives containing naphthalene and fluorine moieties. 2020;(15). doi: 10.1002/ddr.21715 . [DOI] [PubMed] [Google Scholar]
  • 7.Li JJ, Tian Z, Ge X, Xu Z, Feng Y, Liu ZJEJoMC. Design, synthesis, and evaluation of fluorine and Naphthyridine–Based half-sandwich organoiridium/ruthenium complexes with bioimaging and anticancer activity. 2019;163:830–9. doi: 10.1016/j.ejmech.2018.12.021 . [DOI] [PubMed] [Google Scholar]
  • 8.Sharma M, Singh A, Garg N, Kaur N, Singh N. Anticancer SAR Establishment and Novel Accruing Signal Transduction Model of Drug Action Using Biscoumarin Scaffold. Computational Biology and Chemistry. 2019. doi: 10.1016/j.compbiolchem.2019.107104 . [DOI] [PubMed] [Google Scholar]
  • 9.Kessenbrock K, Plaks V, Werb Z. Matrix metalloproteinases: regulators of the tumor microenvironment. Cell. 2010;141(1):52–67. Epub 2010/04/08. doi: 10.1016/j.cell.2010.03.015 ; PubMed Central PMCID: PMC2862057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.Liao H, Zhang L, Lu S, Li W, Dong W. KIFC3 Promotes Proliferation, Migration, and Invasion in Colorectal Cancer via PI3K/AKT/mTOR Signaling Pathway. Front Genet. 2022;13:848926. Epub 2022/07/12. doi: 10.3389/fgene.2022.848926 ; PubMed Central PMCID: PMC9257096. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11.Wagner EF, Nebreda AR. Signal integration by JNK and p38 MAPK pathways in cancer development. Nat Rev Cancer. 2009;9(8):537–49. Epub 2009/07/25. doi: 10.1038/nrc2694 . [DOI] [PubMed] [Google Scholar]
  • 12.Cao Y, Pan R, Xuan W, Wei Y, Liu K, Zhou J, et al. Photo-triggered fluorescent theranostic prodrugs as DNA alkylating agents for mechlorethamine release and spatiotemporal monitoring. Org Biomol Chem. 2015;13(24):6742–8. Epub 2015/05/23. doi: 10.1039/c5ob00500k . [DOI] [PubMed] [Google Scholar]
  • 13.Tuo W, Bouquet J, Taran F, Le Gall T. A FRET probe for the detection of alkylating agents. Chem Commun (Camb). 2019;55(59):8655–8. Epub 2019/07/10. doi: 10.1039/c9cc04391h . [DOI] [PubMed] [Google Scholar]
  • 14.Hueso-Falcon I, Amesty A, Anaissi-Afonso L, Lorenzo-Castrillejo I, Machin F, Estevez-Braun A. Synthesis and biological evaluation of naphthoquinone-coumarin conjugates as topoisomerase II inhibitors. Bioorg Med Chem Lett. 2017;27(3):484–9. Epub 2017/01/04. doi: 10.1016/j.bmcl.2016.12.040 . [DOI] [PubMed] [Google Scholar]
  • 15.Hao SY, Feng SL, Wang XR, Wang Z, Chen SW, Hui L. Novel conjugates of podophyllotoxin and coumarin: Synthesis, cytotoxicities, cell cycle arrest, binding CT DNA and inhibition of Topo IIbeta. Bioorg Med Chem Lett. 2019;29(16):2129–35. Epub 2019/07/07. doi: 10.1016/j.bmcl.2019.06.063 . [DOI] [PubMed] [Google Scholar]
  • 16.Luo G, Chen M, Lyu W, Zhao R, Xu Q, You Q, et al. Design, synthesis, biological evaluation and molecular docking studies of novel 3-aryl-4-anilino-2H-chromen-2-one derivatives targeting ERalpha as anti-breast cancer agents. Bioorg Med Chem Lett. 2017;27(12):2668–73. Epub 2017/05/04. doi: 10.1016/j.bmcl.2017.04.029 . [DOI] [PubMed] [Google Scholar]
  • 17.Kumar A, Sunita P, Jha S, Pattanayak SP. 7,8-Dihydroxycoumarin exerts antitumor potential on DMBA-induced mammary carcinogenesis by inhibiting ERalpha, PR, EGFR, and IGF1R: involvement of MAPK1/2-JNK1/2-Akt pathway. J Physiol Biochem. 2018;74(2):223–34. Epub 2018/02/13. doi: 10.1007/s13105-018-0608-2 . [DOI] [PubMed] [Google Scholar]
  • 18.Yang L, Hu Z, Luo J, Tang C, Zhang S, Ning W, et al. Dual functional small molecule fluorescent probes for image-guided estrogen receptor-specific targeting coupled potent antiproliferative potency for breast cancer therapy. Bioorg Med Chem. 2017;25(13):3531–9. Epub 2017/05/17. doi: 10.1016/j.bmc.2017.05.002 . [DOI] [PubMed] [Google Scholar]
  • 19.Mokale SN, Begum A, Sakle NS, Shelke VR, Bhavale SA. Design, synthesis and anticancer screening of 3-(3-(substituted phenyl) acryloyl)-2H-chromen-2ones as selective anti-breast cancer agent. Biomed Pharmacother. 2017;89:966–72. Epub 2017/03/16. doi: 10.1016/j.biopha.2017.02.089 . [DOI] [PubMed] [Google Scholar]
  • 20.Cui N, Lin DD, Shen Y, Shi JG, Wang B, Zhao MZ, et al. Triphenylethylene-Coumarin Hybrid TCH-5c Suppresses Tumorigenic Progression in Breast Cancer Mainly Through the Inhibition of Angiogenesis. Anticancer Agents Med Chem. 2019;19(10):1253–61. Epub 2019/04/06. doi: 10.2174/1871520619666190404155230 . [DOI] [PubMed] [Google Scholar]
  • 21.Lingaraju GS, Balaji KS, Jayarama S, Anil SM, Kiran KR, Sadashiva MP. Synthesis of new coumarin tethered isoxazolines as potential anticancer agents. Bioorg Med Chem Lett. 2018;28(23–24):3606–12. Epub 2018/11/07. doi: 10.1016/j.bmcl.2018.10.046 . [DOI] [PubMed] [Google Scholar]
  • 22.Singh H, Kumar M, Nepali K, Gupta MK, Saxena AK, Sharma S, et al. Triazole tethered C5-curcuminoid-coumarin based molecular hybrids as novel antitubulin agents: Design, synthesis, biological investigation and docking studies. Eur J Med Chem. 2016;116:102–15. Epub 2016/04/10. doi: 10.1016/j.ejmech.2016.03.050 . [DOI] [PubMed] [Google Scholar]
  • 23.Mokdad-Bzeouich I, Kovacic H, Ghedira K, Chebil L, Ghoul M, Chekir-Ghedira L, et al. Esculin and its oligomer fractions inhibit adhesion and migration of U87 glioblastoma cells and in vitro angiogenesis. Tumour Biol. 2016;37(3):3657–64. Epub 2015/10/16. doi: 10.1007/s13277-015-4209-1 . [DOI] [PubMed] [Google Scholar]
  • 24.Cao D, Liu Y, Yan W, Wang C, Bai P, Wang T, et al. Design, Synthesis, and Evaluation of in Vitro and in Vivo Anticancer Activity of 4-Substituted Coumarins: A Novel Class of Potent Tubulin Polymerization Inhibitors. J Med Chem. 2016;59(12):5721–39. Epub 2016/05/24. doi: 10.1021/acs.jmedchem.6b00158 . [DOI] [PubMed] [Google Scholar]
  • 25.Garazd Y, Garazd M, Lesyk R. Synthesis and evaluation of anticancer activity of 6-pyrazolinylcoumarin derivatives. Saudi Pharm J. 2017;25(2):214–23. Epub 2017/03/28. doi: 10.1016/j.jsps.2016.05.005 ; PubMed Central PMCID: PMC5355548. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 26.Govindaiah P, Dumala N, Grover P, Jaya Prakash M. Synthesis and biological evaluation of novel 4,7-dihydroxycoumarin derivatives as anticancer agents. Bioorg Med Chem Lett. 2019;29(14):1819–24. Epub 2019/05/21. doi: 10.1016/j.bmcl.2019.05.008 . [DOI] [PubMed] [Google Scholar]
  • 27.Luo G, Li X, Zhang G, Wu C, Tang Z, Liu L, et al. Novel SERMs based on 3-aryl-4-aryloxy-2H-chromen-2-one skeleton—A possible way to dual ERalpha/VEGFR-2 ligands for treatment of breast cancer. Eur J Med Chem. 2017;140:252–73. Epub 2017/09/25. doi: 10.1016/j.ejmech.2017.09.015 . [DOI] [PubMed] [Google Scholar]
  • 28.Mohamed TK, Batran RZ, Elseginy SA, Ali MM, Mahmoud AE. Synthesis, anticancer effect and molecular modeling of new thiazolylpyrazolyl coumarin derivatives targeting VEGFR-2 kinase and inducing cell cycle arrest and apoptosis. Bioorg Chem. 2019;85:253–73. Epub 2019/01/15. doi: 10.1016/j.bioorg.2018.12.040 . [DOI] [PubMed] [Google Scholar]
  • 29.Goud NS, Pooladanda V, Mahammad GS, Jakkula P, Gatreddi S, Qureshi IA, et al. Synthesis and biological evaluation of morpholines linked coumarin-triazole hybrids as anticancer agents. Chem Biol Drug Des. 2019;94(5):1919–29. Epub 2019/06/07. doi: 10.1111/cbdd.13578 . [DOI] [PubMed] [Google Scholar]
  • 30.Wang G, Lu M, Yao Y, Wang J, Li J. Esculetin exerts antitumor effect on human gastric cancer cells through IGF-1/PI3K/Akt signaling pathway. Eur J Pharmacol. 2017;814:207–15. Epub 2017/08/30. doi: 10.1016/j.ejphar.2017.08.025 . [DOI] [PubMed] [Google Scholar]
  • 31.Sabt A, Abdelhafez OM, El-Haggar RS, Madkour HMF, Eldehna WM, El-Khrisy E, et al. Novel coumarin-6-sulfonamides as apoptotic anti-proliferative agents: synthesis, in vitro biological evaluation, and QSAR studies. J Enzyme Inhib Med Chem. 2018;33(1):1095–107. Epub 2018/06/27. doi: 10.1080/14756366.2018.1477137 ; PubMed Central PMCID: PMC6022226. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 32.Kurt BZ, Dag A, Dogan B, Durdagi S, Angeli A, Nocentini A, et al. Synthesis, biological activity and multiscale molecular modeling studies of bis-coumarins as selective carbonic anhydrase IX and XII inhibitors with effective cytotoxicity against hepatocellular carcinoma. Bioorg Chem. 2019;87:838–50. Epub 2019/04/20. doi: 10.1016/j.bioorg.2019.03.003 . [DOI] [PubMed] [Google Scholar]
  • 33.Thacker PS, Alvala M, Arifuddin M, Angeli A, Supuran CT. Design, synthesis and biological evaluation of coumarin-3-carboxamides as selective carbonic anhydrase IX and XII inhibitors. Bioorg Chem. 2019;86:386–92. Epub 2019/02/15. doi: 10.1016/j.bioorg.2019.02.004 . [DOI] [PubMed] [Google Scholar]
  • 34.Bonardi A, Falsini M, Catarzi D, Varano F, Di Cesare Mannelli L, Tenci B, et al. Structural investigations on coumarins leading to chromeno[4,3-c]pyrazol-4-ones and pyrano[4,3-c]pyrazol-4-ones: New scaffolds for the design of the tumor-associated carbonic anhydrase isoforms IX and XII. Eur J Med Chem. 2018;146:47–59. Epub 2018/02/07. doi: 10.1016/j.ejmech.2018.01.033 . [DOI] [PubMed] [Google Scholar]
  • 35.Chandak N, Ceruso M, Supuran CT, Sharma PK. Novel sulfonamide bearing coumarin scaffolds as selective inhibitors of tumor associated carbonic anhydrase isoforms IX and XII. Bioorg Med Chem. 2016;24(13):2882–6. Epub 2016/05/04. doi: 10.1016/j.bmc.2016.04.052 . [DOI] [PubMed] [Google Scholar]
  • 36.Wang Y, Cheng FX, Yuan XL, Tang WJ, Shi JB, Liao CZ, et al. Dihydropyrazole derivatives as telomerase inhibitors: Structure-based design, synthesis, SAR and anticancer evaluation in vitro and in vivo. Eur J Med Chem. 2016;112:231–51. Epub 2016/02/24. doi: 10.1016/j.ejmech.2016.02.009 . [DOI] [PubMed] [Google Scholar]
  • 37.Lv N, Sun M, Liu C, Li J. Design and synthesis of 2-phenylpyrimidine coumarin derivatives as anticancer agents. Bioorg Med Chem Lett. 2017;27(19):4578–81. Epub 2017/09/11. doi: 10.1016/j.bmcl.2017.08.044 . [DOI] [PubMed] [Google Scholar]
  • 38.Faisal M, Saeed A, Shahzad D, Fattah TA, Lal B, Channar PA, et al. Enzyme inhibitory activities an insight into the structure-Activity relationship of biscoumarin derivatives. Eur J Med Chem. 2017;141:386–403. Epub 2017/10/17. doi: 10.1016/j.ejmech.2017.10.009 . [DOI] [PubMed] [Google Scholar]
  • 39.Hudacova M, Hamulakova S, Konkolova E, Jendzelovsky R, Vargova J, Sevc J, et al. Synthesis of New Biscoumarin Derivatives, In Vitro Cholinesterase Inhibition, Molecular Modelling and Antiproliferative Effect in A549 Human Lung Carcinoma Cells. Int J Mol Sci. 2021;22(8). Epub 2021/05/01. doi: 10.3390/ijms22083830 ; PubMed Central PMCID: PMC8068036. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 40.Reddy DS, Kongot M, Singh V, Siddiquee MA, Patel R, Singhal NK, et al. Biscoumarin-pyrimidine conjugates as potent anticancer agents and binding mechanism of hit candidate with human serum albumin. Arch Pharm (Weinheim). 2021;354(1):e2000181. Epub 2020/09/19. doi: 10.1002/ardp.202000181 . [DOI] [PubMed] [Google Scholar]
  • 41.Li J, Sui YP, Xin JJ, Du XL, Li JT, Huo HR, et al. Synthesis of biscoumarin and dihydropyran derivatives with promising antitumor and antibacterial activities. Bioorg Med Chem Lett. 2015;25(23):5520–3. Epub 2015/11/03. doi: 10.1016/j.bmcl.2015.10.063 . [DOI] [PubMed] [Google Scholar]
  • 42.Sui YP, Huo HR, Xin JJ, Li J, Li XJ, Du XL, et al. Antibacterial and Antitumor Activities of Biscoumarin and Dihydropyran Derivatives. Molecules. 2015;20(9):17614–26. Epub 2015/09/26. doi: 10.3390/molecules200917614 ; PubMed Central PMCID: PMC6332513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 43.Perumalsamy H, Sankarapandian K, Kandaswamy N, Balusamy SR, Periyathambi D, Raveendiran N. Cellular effect of styrene substituted biscoumarin caused cellular apoptosis and cell cycle arrest in human breast cancer cells. Int J Biochem Cell Biol. 2017;92:104–14. Epub 2017/09/30. doi: 10.1016/j.biocel.2017.09.019 . [DOI] [PubMed] [Google Scholar]
  • 44.Stegh AH. Toward personalized cancer nanomedicine—past, present, and future. Integr Biol (Camb). 2013;5(1):48–65. Epub 2012/08/04. doi: 10.1039/c2ib20104f ; PubMed Central PMCID: PMC3524384. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 45.Malfettone A, Soukupova J, Bertran E, Crosas-Molist E, Lastra R, Fernando J, et al. Transforming growth factor-beta-induced plasticity causes a migratory stemness phenotype in hepatocellular carcinoma. Cancer Lett. 2017;392:39–50. Epub 2017/02/06. doi: 10.1016/j.canlet.2017.01.037 . [DOI] [PubMed] [Google Scholar]
  • 46.Liu H, Liao W, Fan L, Zheng Z, Liu D, Zhang Q-W, et al. Ethanol extract of Ophiorrhiza pumila suppresses liver cancer cell proliferation and migration. Chinese Medicine. 2020;15(1). doi: 10.1186/s13020-020-0291-4 . PubMed Central PMCID: PMC6995237 [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 47.Reymond N, d’Agua BB, Ridley AJ. Crossing the endothelial barrier during metastasis. Nat Rev Cancer. 2013;13(12):858–70. Epub 2013/11/23. doi: 10.1038/nrc3628 . [DOI] [PubMed] [Google Scholar]
  • 48.Sun Y, Liu WZ, Liu T, Feng X, Yang N, Zhou HF. Signaling pathway of MAPK/ERK in cell proliferation, differentiation, migration, senescence and apoptosis. J Recept Signal Transduct Res. 2015;35(6):600–4. Epub 2015/06/23. doi: 10.3109/10799893.2015.1030412 . [DOI] [PubMed] [Google Scholar]
  • 49.Kim ES, Kim MS, Moon A. TGF-beta-induced upregulation of MMP-2 and MMP-9 depends on p38 MAPK, but not ERK signaling in MCF10A human breast epithelial cells. Int J Oncol. 2004;25(5):1375–82. Epub 2004/10/20. . [PubMed] [Google Scholar]
  • 50.Zhang Y, Liu J, Kou J, Yu J, Yu B. DT-13 suppresses MDA-MB-435 cell adhesion and invasion by inhibiting MMP-2/9 via the p38 MAPK pathway. Mol Med Rep. 2012;6(5):1121–5. Epub 2012/08/28. doi: 10.3892/mmr.2012.1047 . [DOI] [PubMed] [Google Scholar]
  • 51.Maeda S, Omata M. Inflammation and cancer: role of nuclear factor-kappaB activation. Cancer Sci. 2008;99(5):836–42. Epub 2008/02/26. doi: 10.1111/j.1349-7006.2008.00763.x . [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 52.Kwon HC, Kim SH, Oh SY, Lee S, Lee JH, Jang JS, et al. Clinicopathologic significance of expression of nuclear factor-kappaB RelA and its target gene products in gastric cancer patients. World J Gastroenterol. 2012;18(34):4744–50. Epub 2012/09/25. doi: 10.3748/wjg.v18.i34.4744 ; PubMed Central PMCID: PMC3442213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 53.Wu Z, He D, Zhao S, Wang H. IL-17A/IL-17RA promotes invasion and activates MMP-2 and MMP-9 expression via p38 MAPK signaling pathway in non-small cell lung cancer. Mol Cell Biochem. 2019;455(1–2):195–206. Epub 2018/12/20. doi: 10.1007/s11010-018-3483-9 . [DOI] [PubMed] [Google Scholar]
  • 54.Teli P, Sethiya A, Gupta S. An Insight View on Synthetic Protocol, Mechanistic and Biological Aspects of Biscoumarin Derivatives. ChemistrySelect. 2019;4:13772–87. Available from: 10.1002/slct.201903632. [DOI] [Google Scholar]

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