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Physico-chemistry and Cytotoxicity of Tenofovir-Loaded Acid Phosphatase-Responsive Chitosan Nanoparticles

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Abstract

This study assesses the in vitro release of tenofovir (TFV)-loaded triphosphate (TPP) cross-linked chitosan nanoparticles (NPs) catalyzed by human prostatic acid phosphatase (hPAP) for 24 h. The physico-chemical characterization of the NPs included particle mean diameter (PMD), zeta potential (ζ), percent drug encapsulation efficiency (% EE), Fourier transform infra-red (FTIR) spectroscopy, powder X-ray diffractometry analysis (PXRD), and drug release kinetics. The first-order rate constant (k) and activation energy (Ea) of the cross-link (TPP) are determined by the integrated rate law and Arrhenius’s equations. The hPAP Michaelis–Menten constant (Km) is determined by the Lineweaver–Burk’s equation. The NP’s safety profile is evaluated on vaginal epithelial cells (VK2/E6E7). The lyophilized drug-loaded NPs’ PMD, ζ, and PDI are 149.97 nm, 4.4 mV, and 0.3, respectively. The % EE after lyophilization is 93.7 ± 4.4%. These NPs released drug at faster rate (63% of TFV within 6 h) under the enzyme’s influence. The similarity and difference factors of drug release profiles (absence vs presence of hPAP) are 56.5 and 40.3, respectively. The hPAP’s Km value of 0.019 mM suggests it has a good affinity for TPP at physiological pH ~ 7.4. The enhanced hydrolysis of TPP or degradation of chitosan NPs is fundamentally due to a decrease of TPP’s activation energy by hPAP. In fact, the Ea value is 22.50 ± 3.06 kJ/mol or 16.33 ± 0.62 kJ/mol in the absence or presence of hPAP, respectively. The NPs are non-cytotoxic to the treated vaginal cell line. These hPAP-responsive NPs are promising topical nanomicrobicides for HIV/AIDS prevention.

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Data Availability

Raw data would be available upon request.

Abbreviations

AM:

Ammonium molybdate tetrahydrate

AT:

Antimonyl tartrate trihydrate

AsA:

Ascorbic acid

Ea :

Activation energy

% EE:

Percent encapsulation efficiency

FTIR:

Fourier transformed infrared

hPAP:

Human prostatic acid phosphatase

Km :

Michaelis constant

MTS:

[3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt

NPs:

Nanoparticles

PMD:

Particle mean diameter

PMS:

Phenazine methosulfate

PXRD:

Powder X-ray diffractometry

SA:

Sodium acetate

TFV:

Tenofovir

TPP:

Triphosphate

Vmax :

Maximal velocity, asymptotic reaction velocity at infinite substrate concentration

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Funding

This work is supported by the award number R01 AI087304, from the National Institute of Allergic and Infectious Diseases (Bethesda, MD, USA).

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Authors and Affiliations

  1. Laboratory of Future Nanomedicines and Theoretical Chronopharmaceutics, Division of Pharmacology and Pharmaceutical Sciences, University of Missouri-Kansas City, Kansas City, Missouri, 64108, USA

    Albert Nguessan Ngo & Bi-Botti Celestin Youan

  2. School of Pharmacy, American University of Health Sciences, Signal Hill, California, 90755, USA

    Albert Nguessan Ngo

  3. College of Medicine, Charles R. Drew University of Medicine and Science, Los Angeles, California, 90059, USA

    Albert Nguessan Ngo

  4. Department of Geosciences, University of Missouri - Kansas City, Kansas City, Missouri, 64110, USA

    James Murowchick

  5. Department of Chemistry, University of Missouri - Kansas City, Kansas City, Missouri, 64110, USA

    Andrea Drew Gounev & Todor K. Gounev

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Contributions

Albert N. Ngo and Bi-Botti Youan (Principal Investigator), were involved in the conception and write-up of the manuscript. The article lists several additional co-authors. Dr. James Murowchick provided XRD analysis of the formulation’s samples at the request of Dr. Ngo and Prof. Youan. Dr Andrea Drew Gounev and Todor K. Gounev provided FTIR analysis at the request of Dr. Ngo and Dr. Youan. We hereby declare that all statements made herein of our own knowledge are true. All authors have read and agreed to the published version of the manuscript.

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Correspondence to Bi-Botti Celestin Youan.

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Ngo, A.N., Murowchick, J., Gounev, A.D. et al. Physico-chemistry and Cytotoxicity of Tenofovir-Loaded Acid Phosphatase-Responsive Chitosan Nanoparticles. AAPS PharmSciTech 24, 143 (2023). https://doi.org/10.1208/s12249-023-02580-1

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