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SN-38, the active metabolite of irinotecan, inhibits the acute inflammatory response by targeting toll-like receptor 4

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Cancer Chemotherapy and Pharmacology Aims and scope Submit manuscript

Abstract

Purpose

Anticancer-drug efficacy seems to involve the direct interaction with host immune cells. Although topoisomerase I (Top I) inhibitors have been suggested to block LPS-evoked inflammation, the interaction between these drugs and toll-like receptor 4 (TLR4) is unaddressed.

Methods

SN-38, the active metabolite of the Top I inhibitor irinotecan, and TLR4 interaction was assessed using the in vitro luciferase nuclear factor-κB reporter assay, neutrophil migration to murine air-pouch, in silico simulation, and the thermal shift assay (TSA). Topotecan was used as a positive anti-inflammatory control.

Results

Non-cytotoxic concentrations of SN-38 attenuated LPS (a TLR4 agonist)-driven cell activation without affecting peptidoglycan (a TLR2 agonist)-activating response. Similarly, topotecan also prevented LPS-induced inflammation. Conversely, increasing concentrations of LPS reversed the SN-38 inhibitory effect. In addition, SN-38 abrogated LPS-dependent neutrophil migration and reduced TNF-α, IL-6, and keratinocyte chemoattractant levels in the air-pouch model, but failed to inhibit zymosan (a TLR2 agonist)-induced cell migration. A two-step molecular docking analysis indicated two potential binding sites for the SN-38 in the MD-2/TLR4 complex, the hydrophobic MD-2 pocket (binding energy of − 8.1 kcal/mol) and the rim of the same molecule (− 6.9 kcal/mol). The topotecan also bound to the MD-2 pocket. In addition, not only the lactone forms, but also the carboxylate conformations of both Top I inhibitors interacted with the MD-2 molecule. Furthermore, the TSA suggested the interaction of SN-38 with MD-2.

Conclusions

Therefore, SN-38 inhibits acute inflammation by blocking LPS-driven TLR4 signaling. This mechanism seems to be shared by other Top I inhibitors.

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Abbreviations

AP-1:

Activator protein 1

COX-2:

Cyclooxygenase-2

DAMP:

Danger-associated molecular patterns

DMSO:

Dimethyl sulfoxide

HTAB:

Hexadecyltrimethyl-ammonium bromide

IL-1β:

Interleukin-1 beta

IL-6:

Interleukin-6

IL-8:

Interleukin-8

iNOS:

Inducible nitric oxide synthase

KC:

Keratinocyte chemoattractant

LPS:

Lipopolysaccharide

MD-2:

Myeloid differentiation-2

MPO:

Myeloperoxidase

MyD88:

Myeloid differentiation primary response 88

NF-κB:

Nuclear factor-κappa B

PAMP:

Pathogen-associated molecular patterns

PBS:

Phosphate buffered saline

PGN:

Peptidoglycan

PMA:

Phorbol 12-myristate 13-acetate

SN-38:

7-Ethyl-10-hydroxycamptothecin

TLR2:

Toll-like receptor 4

TLR4:

Toll-like receptor 2

TLRs:

Toll-like receptors

TNF-α:

Tumor necrosis factor-alpha

Top I:

Topoisomerase I

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Acknowledgements

We are grateful to Giuliana Bertozi, Diva Amabile, Ana Katia dos Santos, Sergio Roberto Rosa, Ieda Regina dos Santos, Maria Silvandira Freire Socorro França, and José Olavo Morais for technical assistance.

Funding

This study was supported by CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico, Grant numbers: 307143/2014–7 and 428354/2016–5), FUNCAP (Fundação Cearense de Apoio ao Desenvolvimento Científico, Grant number: PR2-0101-00054.01.00/15) and CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior). This manuscript is dedicated to the loving memory of Prof. Dr. Ronaldo Albuquerque Ribeiro (in memoriam).

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Authors and Affiliations

  1. Department of Pathology and Forensic Medicine, Faculty of Medicine, Federal University of Ceará, Fortaleza, Brazil

    Deysi Viviana Tenazoa Wong

  2. Department of Pathology, Molecular Biology Laboratory, Cancer Institute of Ceará, Fortaleza, Brazil

    Deysi Viviana Tenazoa Wong, Karla Oliveira Silva & Luis Philipi Carvalho Borges

  3. Department of Physiology and Pharmacology, Drug Research and Development Center, Faculty of Medicine, Federal University of Ceará, Rua Cel Nunes de Melo, 1127, Rodolfo Teófilo, Fortaleza, Ceará, 60430-270, Brazil

    Carlos Wagner Souza Wanderley, Caio Abner Vitorino Gonçalves Leite, Alexia Nathália Brígido Assef, Aurilene Gomes Cajado, Gabriela Loiola Ponte Batista, Rafael Holanda González, Nylane Maria Nunes Alencar, Diego Veras Wilke & Roberto César Pereira Lima-Júnior

  4. Brazilian Biosciences National Laboratory, LNBio, Brazilian Center for Research in Energy and Materials (CNPEM), Campinas, SP, Brazil

    Helder Veras Ribeiro-Filho & Ana Carolina Migliorini Figueira

  5. Center of Health and Biological Sciences, Federal University of Oeste da Bahia, Barreiras, Brazil

    Jonilson Berlink Lima

  6. Department of Pharmacology, School of Medicine of Ribeirão Preto, University of São Paulo, São Paulo, Brazil

    Jonilson Berlink Lima, Thiago Mattar Cunha & Fernando Queiroz Cunha

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  1. Deysi Viviana Tenazoa Wong
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  2. Helder Veras Ribeiro-Filho
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  3. Carlos Wagner Souza Wanderley
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Contributions

Study design: DVTW, CWSW, HVRF, NMNA, FQC, and RCPLJ. Performed the experiments: DVTW, CWSW, HVRF, CAVGL, JBL, ANBA, AGC, GLPB, RHG, KOS, and LPCB. Data analysis: DVTW, CWSW, HVEF, DVW, and TMC. Interpretation of the results: DVTW, NMNA, DVW, TMC, FQC, and RCPLJ. Wrote the paper: DVTW, HVRF, and RCPLJ. All the authors revised and approved the paper.

Corresponding authors

Correspondence to Deysi Viviana Tenazoa Wong or Roberto César Pereira Lima-Júnior.

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The authors declare they have no financial, personal, or professional interests to disclose.

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Wong, D.V.T., Ribeiro-Filho, H.V., Wanderley, C.W.S. et al. SN-38, the active metabolite of irinotecan, inhibits the acute inflammatory response by targeting toll-like receptor 4. Cancer Chemother Pharmacol 84, 287–298 (2019). https://doi.org/10.1007/s00280-019-03844-z

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