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Development of doxorubicin hydrochloride–loaded whey protein nanoparticles and its surface modification with N-acetyl cysteine for triple-negative breast cancer

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Abstract

Limited targeted therapies are available for triple-negative breast cancer (TNBC). Thus, the current research focused on developing a targeted protein nanoparticle for TNBC. First, the doxorubicin hydrochloride (Dox)–loaded genipin-crosslinked whey protein nanoparticles (WD) were prepared and optimised by the QbD method using BBD. The hydrodynamic diameter of WD was found to be 364.38 ± 49.23 nm, zeta potential −27.59 ± 1.038 mV, entrapment 63.03 ± 3.625% and Dox loading was found to be 1.419 ± 0.422%. The drug recovery after 18 months of storage was 69%. Then, it was incubated with NAC to obtain modified WD (CyWD). WD followed first-order release kinetics, whereas CyWD followed the Higuchi model. Hemagglutination and hemolysis were not found qualitatively in WD and CyWD. Upon injecting the nanoformulations to 4T1-induced mice, the highest efficacy was found to be in CyWD followed by WD and Dox injection. Upon histopathological observance, it was found that the CyWD group gave the most significant damage to the 4T1 tumour tissue. Thus, NAC-modified protein nanoparticles carrying chemotherapeutic agents can be an excellent targeted therapeutic system against TNBC.

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All data and resource materials are available with the correspondence and first author.

Abbreviations

ATCC:

American Type Culture Collection

BBD:

Box Behnken Design

CPCSEA:

Committee for the Purpose of Control and Supervision of Experiments on Animals

DMSO:

Dimethyl sulfoxide

Dox:

Doxorubicin hydrochloride

EDTA:

Ethylenediamine tetraacetic acid

FBS:

Fetal bovine serum

FITC:

Fluorescein isothiocyanate

MWCO:

Molecular weight cut-off

MTT:

3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide

NCCS:

National Centre for Cell Science

PBS:

Phosphate buffered saline

QbD:

Quality by Design

RBC:

Red blood cells

TNBC:

Triple-negative breast cancer

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Acknowledgements

Deshpande Labs, Bhopal, India for in-vivo studies (CPCSEA approved); University Sophisticated Instrumentation Centre-Babasaheb Bhimrao Ambedkar University: Confocal microscopy; Dr Dinesh Kumar and his scholars (Ritu Raj and Umesh Kumar), Center of Biomedical Research, Lucknow, India – centrifuge and lyophilisation.

Funding

Indian Council of Medical Research (No. 3/2/2/42/2020-NCD-III), dated 17.06.2021, awarded to Samipta Singh (SRF) and Prof. Shubhini A. Saraf (Principal Investigator). UGC-Non NET (2016–2020) and ICMR-SRF (2021 onwards) fellowship for financial support to the first author.

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Authors and Affiliations

  1. Department of Pharmaceutical Sciences, Babasaheb Bhimrao Ambedkar University, Vidya Vihar, Raebareily Road, Lucknow, India

    Samipta Singh, Priyanka Maurya, Soniya Rani, Nidhi Mishra, Raquibun Nisha, Priya Singh & Shubhini A. Saraf

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  1. Samipta Singh
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  7. Shubhini A. Saraf
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Contributions

Prof. Shubhini A. Saraf designed and supervised; Samipta Singh: wrote and performed experiments; Priyanka Maurya, Raquibun Nisha, Priya Singh, Soniya Rani and Nidhi Mishra: research co-ordination.

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Correspondence to Shubhini A. Saraf.

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Animals utilised in the study were approved by the Institutional Animal Ethics Committee (IAEC no. DL/SS/11/20/615).

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Singh, S., Maurya, P., Rani, S. et al. Development of doxorubicin hydrochloride–loaded whey protein nanoparticles and its surface modification with N-acetyl cysteine for triple-negative breast cancer. Drug Deliv. and Transl. Res. 12, 3047–3062 (2022). https://doi.org/10.1007/s13346-022-01169-8

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