Microtubule-Targeting NAP Peptide-Ru(II)-polypyridyl Conjugate As a Bimodal Therapeutic Agent for Triple Negative Breast Carcinoma
Triple-negative breast cancer (TNBC) poses significant treatment challenges due to its high metastasis, heterogeneity, and poor biomarker expression. The N-terminus of an octapeptide NAPVSIPQ (<b>NAP</b>) was covalently coupled to a carboxylic acid derivative of Ru(2,2'-bipy)<sub>3</sub><sup>2+</sup> (<b>Rubpy</b>) to synthesize an N-stapled short peptide-Rubpy conjugate (<b>Ru-NAP</b>). This photosensitizer (PS) was utilized to treat TNBC through microtubule (MT) targeted chemotherapy and photodynamic therapy (PDT). <b>Ru-NAP</b> formed more elaborate molecular aggregates with fibrillar morphology as compared to <b>NAP</b>. A much higher binding affinity of <b>Ru-NAP</b> over <b>NAP</b> toward β-tubulin (<i>K</i><sub>Ru-NAP</sub>: (6.8 ± 0.55) × 10<sup>6</sup> M<sup>-1</sup>; <i>K</i><sub>NAP</sub>: (8.2 ± 1.1) × 10<sup>4</sup> M<sup>-1</sup>) was observed due to stronger electrostatic interactions between the MT with an average linear charge density of ∼85 e/nm and the cationic <b>Rubpy</b> part of <b>Ru-NAP</b>. This was also supported by docking, simulation, and appropriate imaging studies. <b>Ru-NAP</b> promoted serum stability, specific binding of <b>NAP</b> to the E-site of the β<sup>III</sup>-tubulin followed by the disruption of the MT network, and effective singlet oxygen generation in TNBC cells (MDA-MB-231), causing cell cycle arrest in the G2/M phase and triggering apoptosis. Remarkably, MDA-MB-231 cells were more sensitive to <b>Ru-NAP</b> compared to noncancerous human embryonic kidney (HEK293 cells) when exposed to light (<sup>Light</sup>IC<sub>50</sub><sup>Ru-NAP</sup>[HEK293]: 17.2 ± 2.5 μM, compared to <sup>Light</sup>IC<sub>50</sub><sup>Ru-NAP</sup>[MDA-MB-231]: 32.5 ± 7.8 nM, <sup>Dark</sup>IC<sub>50</sub><sup>Ru-NAP</sup>[HEK293]: > 80 μM, compared to <sup>Dark</sup>IC<sub>50</sub><sup>Ru-NAP</sup>[MDA-MB-231]: 2.9 ± 0.5 μM). <b>Ru-NAP</b> also effectively inhibited tumor growth in MDA-MB-231 xenograft models in nude mice. Our findings provide strong evidence that <b>Ru-NAP</b> has a potential therapeutic role in TNBC treatment.
Authors
- Anitha Ethirajan (ORCID: https://orcid.org/0000-0002-2264-2536)
- Uttam Pal (ORCID: https://orcid.org/0000-0003-2110-4610)
- Arpan Bhattacharyya (ORCID: https://orcid.org/0000-0002-7933-6441)
- Batakrishna Jana (ORCID: https://orcid.org/0000-0003-2944-0109)
- Amitava Das (ORCID: https://orcid.org/0000-0003-3666-1743)
- Surajit Barman (ORCID: https://orcid.org/0000-0003-3584-4716)
- Benu Brata Das (ORCID: https://orcid.org/0000-0003-2519-7105)
- Sangheeta Bhattacharjee (ORCID: https://orcid.org/0000-0001-9490-1376)
- Atin Chatterjee
- Sandip Sarkar (ORCID: https://orcid.org/0000-0002-0813-5773)
- Raviranjan Pandey (ORCID: https://orcid.org/0009-0002-2709-8856)
Institutions
- Indian Institute of Science Education and Research Kolkata (IN)
- Indian Association for the Cultivation of Science (IN)
- MRC Laboratory of Molecular Biology (GB)
- Institute of Molecular Functional Materials (CN)
- S.N. Bose National Centre for Basic Sciences (IN)
- Hasselt University (BE)
Publication Details
- Journal
- Journal of the American Chemical Society
- Published
- 2024-12-26
- DOI
- https://doi.org/10.1021/jacs.4c11820
- Citations
- 12
- Primary Topic
- Click Chemistry and Applications
- Type
- article
- Field-Weighted Citation Impact
- 3.00
Funders
- Indian Council of Medical Research
- Council of Scientific and Industrial Research, India
- Ministry of Education, India
- Universiteit Hasselt
- Indian Institute of Science Education and Research Mohali
- Indian Institute of Science Education and Research Kolkata
- Science and Engineering Research Board
- Board of Research in Nuclear Sciences