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IITZ-01, a novel potent lysosomotropic autophagy inhibitor, has single-agent antitumor efficacy in triple-negative breast cancer in vitro and in vivo

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Abstract

Autophagy is a homeostatic process that recycles damaged organelles and long-lived proteins by delivering them in double-membrane vesicles to lysosomes for degradation. Autophagy has a prominent role in survival, proliferation, and resistance of tumors in metabolic and chemotherapeutic stress conditions. Clinical trials with chloroquine—a known autophagy inhibitor—were unable to achieve complete autophagy inhibition in vivo, warranting the search for more potent autophagy inhibitors. In a process of exploring the mechanism of action of previously identified cytotoxic s-triazine analogs, we discovered that both IITZ-01 and IITZ-02 act as potent autophagy inhibitors. Treatment with these compounds resulted in the vacuolated appearance of cells due to their specific accumulation in lysosomes. In addition, these basic compounds also deacidify lysosomes as evidenced by the decrease in lysotracker red staining and inhibit maturation of lysosomal enzymes leading to lysosomal dysfunction. IITZ-01 and IITZ-02 enhance autophagosome accumulation but inhibit autophagosomal degradation by impairing lysosomal function, finally resulting in the inhibition of autophagy. Interestingly, compound IITZ-01 exhibited more than 10-fold potent autophagy inhibition along with 12- to 20-fold better cytotoxic action than CQ. IITZ-01 and IITZ-02 also abolished mitochondrial membrane potential and triggered apoptosis through the mitochondria-mediated pathway. Furthermore, IITZ-01 and IITZ-02 displayed potent antitumor action in vivo through autophagy inhibition and apoptosis induction in MDA-MB-231 breast cancer xenograft model with IITZ-01 exhibiting superior anticancer efficacy. Overall, these data demonstrate that IITZ-01 is potent autophagy inhibitor with single-agent anticancer activity and awaits further preclinical development as potential anticancer therapeutic.

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Acknowledgements

GL and DT are thankful to Department of Pharmaceuticals, Ministry of Chemicals and Fertilizers, Government of India, and Director of NIPER-Hyderabad and NIPER-Guwahati, for providing fellowship and research activity to the authors. We also acknowledge Dr M Lakshman, Sri Venkateshwara University College of Veterinary Sciences, for his help in processing and acquisition of transmission electron microscopy images. We thank Jaya Lakshmi, Veera Bhadra Swami, and Vasu Penugurti for their technical guidance and support.

Compliance with ethical standards

All the animal experimentation were carried out in compliance with guidelines of IAEC, NIPER-Hyderabad, India.

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  1. Naidu G. M. Vegi

    Present address: Department of Pharmacology and Toxicology, National Institute of Pharmaceutical Education and Research, Guwahati, Assam, India

Authors and Affiliations

  1. Department of Pharmacology and Toxicology, National Institute of Pharmaceutical Education and Research, Hyderabad, TS, India

    Lalita Guntuku, Dinesh Thummuri & Naidu G. M. Vegi

  2. Crop Protection Chemicals Division, CSIR-Indian Institute of Chemical Technology, Hyderabad, TS, India

    Jagadeesh Kumar Gangasani & Jayathirtha Rao Vaidya

  3. Academy of Scientific and Innovative Research (AcSIR), Training and Development Complex, CSIR Campus, CSIR Road, Chennai, TN, India

    Jagadeesh Kumar Gangasani & Jayathirtha Rao Vaidya

  4. National Centre for Mass Spectrometry, CSIR-Indian Institute of Chemical Technology, Hyderabad, TS, India

    Roshan M. Borkar & Srinivas Ragampeta

  5. Department of Biochemistry, School of Life Sciences, University of Hyderabad, Hyderabad, TS, India

    Bramanandam Manavathi

  6. Medicinal Chemistry and Pharmacology Division, CSIR-Indian Institute of Chemical Technology (IICT), Hyderabad, TS, India

    Ramakrishna Sistla

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  1. Lalita Guntuku
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Guntuku, L., Gangasani, J.K., Thummuri, D. et al. IITZ-01, a novel potent lysosomotropic autophagy inhibitor, has single-agent antitumor efficacy in triple-negative breast cancer in vitro and in vivo. Oncogene 38, 581–595 (2019). https://doi.org/10.1038/s41388-018-0446-2

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