Discovery of dual inhibitors of topoisomerase I and Cyclooxygenase-2 for colon cancer therapy

https://doi.org/10.1016/j.ejmech.2022.114560Get rights and content

Highlights

  • Tolfenamic acid derivatives as Topo I and COX-2 dual inhibitors were reported.

  • W10 exhibited improved water solubility than precursor.

  • W10 showed potential anti-colon cancer activity in vitro and vivo.

Abstract

Novel tolfenamic acid derivatives based on the structure of I-1 were designed and synthesized to improve its poor target inhibition and solubility. Among them, W10 was identified as a potent dual-target inhibitor of Topo I (IC50 = 0.90 ± 0.17 μM) and COX-2 (IC50 = 2.31 ± 0.07 μM) with improved water solubility (32.33 μg/mL). Moreover, W10 also exhibited fairly potent anti-proliferative and pro-apoptosis activity via the mitochondrial pathway, as well as suppressed aberrant NF-κB/IκB activation in colon cancer cells in vitro. Additionally, W10 possessed favorable pharmacokinetic properties and excellent antitumor effects in vivo. In general, our study has demonstrated the potency of a novel Topo I/COX-2 dual inhibitor, which can potentially be developed into a chemotherapeutic candidate for colon cancer.

Introduction

Colon cancer is one of the common malignancies with the third-highest morbidity and second-highest mortality worldwide in 2020 [1,2]. Chemotherapy plays an important role throughout the treatment of colon cancer especially for advanced colon cancer [3]. The clinical first-line chemotherapeutic drugs for advanced colon cancer are mainly 5-FU and oxaliplatin, both of which are limited by their non-selective cytotoxicity and serious side effects [4].

Given the progression characterized by alteration in multiple molecular pathways, developing novel chemotherapeutic drugs against colon cancer became challenging. Targeting multiple highly expressed proteins can provide ideas for the development of such drugs.

Topoisomerase I (Topo I) is overexpressed in around 50% of primary colon cancer compared with normal tissues, closely associated with the rapid proliferation of tumor cells [5,6]. Irinotecan is a Topo I inhibitor possessing greater water solubility than precursor camptothecin with superior anticancer efficiency and comparatively low toxicity in vivo [[7], [8], [9]]. Cyclooxygenase-2 (COX-2) is another highly expressed protein in colon cancer for there are about 40%–50% of colon adenomas and 80%–90% of colon cancer tissues exhibit overexpression of COX-2 [10,11]. Overexpressed COX-2 contributes to cancer progression by inhibiting apoptosis and promoting angiogenesis and invasion [[12], [13], [14], [15]]. It was verified that the chemotherapy regimen combining irinotecan and celecoxib could enhance anticancer activity and decrease toxicity, while this combined therapy involved drug-drug interaction which prevented their clinical application [[16], [17], [18], [19]]. Development of pharmaceutical molecules with dual inhibition of Topo I and COX-2 could not only circumvent the potential risks of combined administration but also provide an alternative for colon cancer therapy drug discovery.

In the current study, we focused on the development of pharmaceutical molecules with both Topo I and COX-2 inhibitory activities. A series of N-phenyl-2-(phenylamino)benzamide derivatives with potential dual inhibitory effects of Topo I and COX-2 based on pseudocycle strategy and combination principle were developed in the previous study [20]. But, all of them suffered from poor efficiency (IC50 > 30 μM for Topo I and COX-2) and undesirable water solubility. Encouraged by several successful works that improve target inhibition and water solubility by introducing polar groups such as sulfamide and ethylene glycol derivatives, we decided to apply this strategy to improve target inhibition and aqueous solubility of dual inhibitors [[21], [22], [23], [24], [25]]. We selected I-1 (IC50 = 33.64 ± 3.99 μM for Topo I, IC50 = 33.61 ± 1.15 μM for COX-2 and water solubility <1 μg/mL) with relatively high activity as the scaffold for modification. Since the 4-OMe of I-1 has not been explored adequately even though it had great potency for structure modification, we choose 4-OMe as the modified site. We designed and synthesized a series of novel tolfenamic acid derivatives modified with polar groups such as sulfamide and ethylene glycol derivatives to obtain highly potent Topo I and COX-2 dual inhibitors for colon cancer therapy.

Section snippets

Chemistry

In order to improve the pharmacological activity, we proceeded a systematic modification to I-1. The synthetic protocols were established in Scheme 1, Scheme 2, Scheme 3, Scheme 4. Generally, amines 4, 7, 11 and 13 were prepared through nucleophilic substitution or condensation reaction and catalytic hydrogenation. Sulfanilamide and 4-aminobenzamide were commercially purchased. Tolfenamic acid reacted in a condensation reaction with amines to obtain W1 – W6, W9, W11 – W19, intermediates I1 and

Conclusion

Topo I and COX-2 are highly expressed in various types of colon cancer and are closely related to tumor progression. Development of drugs targeting both Topo I and COX-2 would hold great promise in colon cancer treatment. In our report, a collection of novel tolfenamic acid derivatives were designed and synthesized to improve the efficiency of dual-targeted inhibition as well as aqueous solubility. The results of target inhibition assay showed that most compounds had better activity compared to

Chemistry

General information. All reagents are analytical grade or chemical grade. Bruker spectrometer (Bruker Company, Germany) was used to record nuclear magnetic resonance (proton NMR 400 MHz and carbon NMR 101 MHz). Using DMSO‑d6, CDCl3 or Methanol-d4 as deuterated solvents and with tetramethylsilane (TMS) as the internal reference. Coupling constants Hz and distance unit ppm are selected as parameters to represent the values of the chemical shifts. Mass spectrometry and high-resolution mass

Author contributions

§Hu XL and Li JF contributed equally to this work. The manuscript was written through contributions of all authors. All authors have given approval to the final version of the manuscript.

Notes

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Declaration of competing interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgments

Thanks to the instrument platform of the School of Chemistry and Chemical Engineering, School of Pharmacy and School of Basic Medical Sciences of Lanzhou University. This research was supported by the Recruitment Program of Global Experts (1000 Talents Program); Gansu Province Science Foundation for Distinguished Young Scholars (20JR5RA304).

References (54)

  • H.Y. Li et al.

    The 5-substituted piperazine as a novel secondary pharmacophore greatly improving the physical properties of urea-based inhibitors of soluble epoxide hydrolase

    Bioorg. Med. Chem.

    (2006)
  • L.I. Pilkington et al.

    Development, synthesis and biological investigation of a novel class of potent PC-PLC inhibitors

    Eur. J. Med. Chem.

    (2020)
  • F. Pin et al.

    Intramolecular N-aza-amidoalkylation in association with Witkop–Winterfeldt oxidation as the key step to synthesize Luotonin-A analogues

    Tetrahedron

    (2011)
  • R. Ulus et al.

    Synthesis of novel acridine and bis acridine sulfonamides with effective inhibitory activity against the cytosolic carbonic anhydrase isoforms II and VII

    Bioorg. Med. Chem.

    (2013)
  • K.R.A. Abdellatif et al.

    Thiohydantoin derivatives incorporating a pyrazole core: design, synthesis and biological evaluation as dual inhibitors of topoisomerase-I and cycloxygenase-2 with anti-cancer and anti-inflammatory activities

    Bioorg. Chem.

    (2019)
  • A. Soleimani et al.

    Role of the NF-kappaB signaling pathway in the pathogenesis of colorectal cancer

    Gene

    (2020)
  • T. Miao et al.

    Induction, fixation and recovery of self-organized helical superstructures in achiral liquid crystalline polymer

    Polym. Chem.

    (2021)
  • R. Ding et al.

    Eeyarestatin I derivatives with improved aqueous solubility

    Bioorg. Med. Chem. Lett.

    (2016)
  • L.I. Pilkington et al.

    Development, synthesis and biological investigation of a novel class of potent PC-PLC inhibitors

    Eur. J. Med. Chem.

    (2020)
  • H. Yang et al.

    Optimization of WZ4003 as NUAK inhibitors against human colorectal cancer

    Eur. J. Med. Chem.

    (2021)
  • A. Yamani et al.

    Discovery and optimization of novel pyrazole-benzimidazole CPL304110, as a potent and selective inhibitor of fibroblast growth factor receptors FGFR (1-3)

    Eur. J. Med. Chem.

    (2021)
  • H. Nakano et al.

    Development of a potent and selective FLT3 kinase inhibitor by systematic expansion of a non-selective fragment-screening hit

    Bioorg. Med. Chem. Lett.

    (2016)
  • Y. Long et al.

    Discovery of novel 4-azaaryl-N-phenylpyrimidin-2-amine derivatives as potent and selective FLT3 inhibitors for acute myeloid leukaemia with FLT3 mutations

    Eur. J. Med. Chem.

    (2021)
  • H. Sung et al.

    Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 Countries

    Ca - Cancer J. Clin.

    (2021)
  • J. Ferlay et al.

    GLOBOCAN 2020 annexes

  • M. Ikeguchi et al.

    Topoisomerase I expression in tumors as a biological marker for CPT-11 chemosensitivity in patients with colorectal cancer

    Surg. Today

    (2011)
  • D. Vallbohmer et al.

    Molecular determinants of irinotecan efficacy

    Int. J. Cancer

    (2006)
  • Cited by (2)

    • Small-molecule drugs of colorectal cancer: Current status and future directions

      2024, Biochimica et Biophysica Acta - Molecular Basis of Disease
    1

    Hu XL and Li JF contributed equally to this work.

    View full text