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Uptake and efflux of FL118 and two FL118 derivatives in 3D cell model

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Abstract

Drug uptake and efflux are two of the critical factors required in order to be able to define drug efficacy. This study aims to investigate cytotoxicity and uptake mechanisms of two FL118 analogues (7-Q20 and val-FL118) in parallel with FL118 in three-dimensional multi-cellular spheroids model. The influence of compound concentration, time, temperature, cell lines, and the inhibitors of P-gp, BCRP and LAT1 on drug uptake and efflux were investigated. In vitro cytotoxicity studies revealed that FL118, 7-Q20 and val-FL118 exhibited sensitive cytotoxicity to the HCT-116 cell line and the water-soluble compound 7-Q20 showed the lowest IC50. Cellular uptake and efflux of FL118 was independent of efflux pump proteins. Uptake and efflux of 7-Q20 were affected by P-gp, which was one of reasons that caused a lower uptake at 37 °C than at 4 °C. The carrier protein LAT1 played a role in the cellular intakes of val-FL118. These findings provided basic information for FL118 and the two novel FL118 derivatives for further development.

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Abbreviations

FL118:

10,11-Methylenedioxy-20(S)-camptothecin

7-Q20:

7-p-Trifluoromethylphenyl-10,11-methylenedioxy-20(S)-camptothecin

val-FL118:

10,11-Methylenedioxy-20 (S)-valine-camptothecin

MTT:

3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide

3D:

Three dimension

P-gp:

P-glycoprotein

BCRP:

Breast cancer resistant protein

LAT1:

L-type amino acid transporter 1

References

  • Bonnier F et al (2015) Cell viability assessment using the Alamar blue assay: a comparison of 2D and 3D cell culture models. Toxicol In Vitro 29:124–131

    Article  CAS  PubMed  Google Scholar 

  • Casey A, Gargotti M, Bonnier F, Byrne HJ (2016) Chemotherapeutic efficiency of drugs in vitro: comparison of doxorubicin exposure in 3D and 2D culture matrices. Toxicol In Vitro 33:99–104

    Article  CAS  PubMed  Google Scholar 

  • Chitrangi S, Nair P, Khanna A (2017) 3D engineered In vitro hepatospheroids for studying drug toxicity and metabolism. Toxicol In Vitro 38:8–18

    Article  CAS  PubMed  Google Scholar 

  • Fan MZ, Adeola O, Mcburney MI, Cheeseman CI (1998) Kinetic analysis of l-glutamine transport into porcine jejunal enterocyte brush-border membrane vesicles. Comp Biochem Physiol Part A Mol Integr Physiol 121:411–422

    Article  CAS  Google Scholar 

  • Fang Y, Liang F, Liu K, Qaiser S, Pan S, Xu X (2018) Structure characteristics for intestinal uptake of flavonoids in Caco-2 cells. Food Res Int 105:353–360

    Article  CAS  PubMed  Google Scholar 

  • Howes AL, Richardson RD, Finlay D, Vuori K (2014) 3-Dimensional culture systems for anti-cancer compound profiling and high-throughput screening reveal increases in EGFR inhibitor-mediated cytotoxicity compared to monolayer culture systems. PLoS ONE 9:e108283

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kinne A, Schülein R, Krause G (2011) Primary and secondary thyroid hormone transporters. Thyroid Res 4:S7

    Article  PubMed  PubMed Central  Google Scholar 

  • Li F (2014) Anticancer drug FL118 is more than a survivin inhibitor: where is the Achilles’ heel of cancer? Am J Cancer Res 4:304–311

    PubMed  PubMed Central  Google Scholar 

  • Li F et al (2017) Topoisomerase I (Top1): a major target of FL118 for its antitumor efficacy or mainly involved in its side effects of hematopoietic toxicity? Am J Cancer Res 7:370–382

    CAS  PubMed  PubMed Central  Google Scholar 

  • Liang QQ, Li YS (2013) A rapid and accurate method for determining protein content in dairy products based on asynchronous-injection alternating merging zone flow-injection spectrophotometry. Food Chem 141:2479–2485

    Article  CAS  PubMed  Google Scholar 

  • Ling X, Cao S, Cheng Q, Keefe JT, Rustum YM, Li F (2012) A novel small molecule FL118 that selectively inhibits survivin, Mcl-1, XIAP and cIAP2 in a p53-independent manner, shows superior antitumor activity. PLoS ONE 7:e45571

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ling X, Liu X, Zhong K, Smith N, Prey J, Li F (2015) FL118, a novel camptothecin analogue, overcomes irinotecan and topotecan resistance in human tumor xenograft models. Am J Transl Res 7:1765–1781

    CAS  PubMed  PubMed Central  Google Scholar 

  • Liu J-A, Guo X-P, Liang S, An F, Shen H-Y, Xu Y-J (2015) Regioselective synthesis of 5′-amino acid esters of some nucleosides via orthogonal protecting protocol. Tetrahedron 71:1409–1412

    Article  CAS  Google Scholar 

  • Luo FR, Paranjpe Guo A, Rubin E, Sinko P (2002) Intestianl transport of irinotecon in Caco-2 cells and MDCK11 cells overexpressing efflux transporters PGP, cMOAT, and MRP1. Drug Metab Distrib 30:763–770

    Article  CAS  Google Scholar 

  • Mehta G, Hsiao AY, Ingram M, Luker GD, Takayama S (2012) Opportunities and challenges for use of tumor spheroids as models to test drug delivery and efficacy. J Control Release 164:192–204

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nath S, Devi GR (2016) Three-dimensional culture systems in cancer research: focus on tumor spheroid model. Pharmacol Ther 163:94–108

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pullakhandam R, Nair MK, Kasula S, Kilari S, Thippande TG (2008) Ferric reductase activity of low molecular weight human milk fraction is associated with enhanced iron solubility and uptake in Caco-2 cells. Biochem Biophys Res Commun 374:369–372

    Article  CAS  PubMed  Google Scholar 

  • Wadkins RM et al (1999) Water soluble 20(S)-glycinate esters of 10,11-methylenedioxycamptothecins are highly active against human breast cancer xenografts. Cancer Res 59:3424–3428

    CAS  PubMed  Google Scholar 

  • Wang W, Clarkson TW, Ballatori N (2000) Gamma-glutamyl transpeptidase and l-cysteine regulate methylmercury uptake by HepG2 cells, a human hepatoma cell line. Toxicol Appl Pharmacol 168:72–78

    Article  CAS  PubMed  Google Scholar 

  • Westover D et al (2015) FL118, a novel camptothecin derivative, is insensitive to ABCG2 expression and shows improved efficacy in comparison with irinotecan in colon and lung cancer models with ABCG2-induced resistance. Mol Cancer 14:92–103

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yang Z et al (2018) FL118, a novel camptothecin analogue, suppressed migration and invasion of human breast cancer cells by inhibiting epithelial-mesenchymal transition via the Wnt/beta-catenin signaling pathway. Biosci Trends 12:40–46

    Article  CAS  PubMed  Google Scholar 

  • Zhao X et al (2014) Design and synthesis of new 7-(N-substituted-methyl)-camptothecin derivatives as potent cytotoxic agents. Bioorg Med Chem Lett 24:3850–3853

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhu G et al (2017) Design, synthesis and potent cytotoxic activity of novel 7-(N-[(substitutedsulfonyl)piperazinyl]-methyl)-camptothecin derivatives. Bioorg Med Chem Lett 27:1750–1753

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We thank Ms. Amanda Hess for editorial proofreading of the manuscript before submission and final publication.

Funding

This work was financially supported by Qianjiang Talents Project in Zhejiang Province.

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Authors and Affiliations

  1. Collaborative Innovation Center of Yangtze River Region Green Pharmaceuticals, Zhejiang University of Technology, No. 18 Chaowang Road, Hangzhou City, 310014, China

    Leilei Zhou & Qingyong Li

  2. College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou, China

    Leilei Zhou, Qi Weng, Yixin Zheng, Yuqin Zhou & Qingyong Li

  3. Department of Pharmacology and Therapeutics, Roswell Park Comprehensive Cancer Center, Buffalo, NY, USA

    Fengzhi Li

Authors
  1. Leilei Zhou
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  2. Qi Weng
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  3. Yixin Zheng
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  4. Yuqin Zhou
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  5. Qingyong Li
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  6. Fengzhi Li
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Contributions

All authors contributed to the study conception and design. Material preparation was performed by YZ and YZ, theoretical advice was provided by QL and FL, data collection and analysis were performed by LZ and QW. The first draft of the manuscript was written by LZ. All authors read and approved the final manuscript.

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Correspondence to Qingyong Li.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Zhou, L., Weng, Q., Zheng, Y. et al. Uptake and efflux of FL118 and two FL118 derivatives in 3D cell model. Cytotechnology 71, 785–795 (2019). https://doi.org/10.1007/s10616-019-00322-5

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  • DOI: https://doi.org/10.1007/s10616-019-00322-5

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