Synthesis, characterization, and mechanistic studies of a gold nanoparticle–amphotericin B covalent conjugate with enhanced antileishmanial efficacy and reduced cytotoxicity
Prakash Kumar,1 Pushkar Shivam,*,2 Saptarshi Mandal,*,3 Pragya Prasanna,1 Saurabh Kumar,1 Surendra Rajit Prasad,1 Ashish Kumar,4 Prolay Das,3 Vahab Ali,5 Shubhankar Kumar Singh,2 Debabrata Mandal11Department of Biotechnology, National Institute of Pharmaceutical Education and Research (NIPER) &n...
Guardado en:
| Autores principales: | , , , , , , , , , , |
|---|---|
| Formato: | article |
| Lenguaje: | EN |
| Publicado: |
Dove Medical Press
2019
|
| Materias: | |
| Acceso en línea: | https://doaj.org/article/303109f71b1a4ab28a430db566d3a540 |
| Etiquetas: |
Agregar Etiqueta
Sin Etiquetas, Sea el primero en etiquetar este registro!
|
| id |
oai:doaj.org-article:303109f71b1a4ab28a430db566d3a540 |
|---|---|
| record_format |
dspace |
| spelling |
oai:doaj.org-article:303109f71b1a4ab28a430db566d3a5402021-12-02T11:54:04ZSynthesis, characterization, and mechanistic studies of a gold nanoparticle–amphotericin B covalent conjugate with enhanced antileishmanial efficacy and reduced cytotoxicity1178-2013https://doaj.org/article/303109f71b1a4ab28a430db566d3a5402019-08-01T00:00:00Zhttps://www.dovepress.com/synthesis-characterization-and-mechanistic-studies-of-a-gold-nanoparti-peer-reviewed-article-IJNhttps://doaj.org/toc/1178-2013Prakash Kumar,1 Pushkar Shivam,*,2 Saptarshi Mandal,*,3 Pragya Prasanna,1 Saurabh Kumar,1 Surendra Rajit Prasad,1 Ashish Kumar,4 Prolay Das,3 Vahab Ali,5 Shubhankar Kumar Singh,2 Debabrata Mandal11Department of Biotechnology, National Institute of Pharmaceutical Education and Research (NIPER) – Hajipur, Vaishali, India; 2Department of Microbiology, Rajendra Memorial Research Institute of Medical Sciences, Patna, India; 3Department of Chemistry, Indian Institute of Technology Patna, Patna, India; 4Department of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences, Patna, India; 5Department of Biochemistry, Rajendra Memorial Research Institute of Medical Sciences, Patna, India*These authors contributed equally to this workBackground: Amphotericin B (AmB) as a liposomal formulation of AmBisome is the first line of treatment for the disease, visceral leishmaniasis, caused by the parasite Leishmania donovani. However, nephrotoxicity is very common due to poor water solubility and aggregation of AmB. This study aimed to develop a water-soluble covalent conjugate of gold nanoparticle (GNP) with AmB for improved antileishmanial efficacy and reduced cytotoxicity.Methods: Citrate-reduced GNPs (∼39 nm) were functionalized with lipoic acid (LA), and the product GNP-LA (GL ∼46 nm) was covalently conjugated with AmB using carboxyl-to-amine coupling chemistry to produce GNP-LA-AmB (GL-AmB ∼48 nm). The nanoparticles were characterized by dynamic light scattering, transmission electron microscopy (TEM), and spectroscopic (ultraviolet–visible and infrared) methods. Experiments on AmB uptake of macrophages, ergosterol depletion of drug-treated parasites, cytokine ELISA, fluorescence anisotropy, flow cytometry, and gene expression studies established efficacy of GL-AmB over standard AmB.Results: Infrared spectroscopy confirmed the presence of a covalent amide bond in the conjugate. TEM images showed uniform size with smooth surfaces of GL-AmB nanoparticles. Efficiency of AmB conjugation was ∼78%. Incubation in serum for 72 h showed <7% AmB release, indicating high stability of conjugate GL-AmB. GL-AmB with AmB equivalents showed ∼5-fold enhanced antileishmanial activity compared with AmB against parasite-infected macrophages ex vivo. Macrophages treated with GL-AmB showed increased immunostimulatory Th1 (IL-12 and interferon-γ) response compared with standard AmB. In parallel, AmB uptake was ∼5.5 and ∼3.7-fold higher for GL-AmB-treated (P<0.001) macrophages within 1 and 2 h of treatment, respectively. The ergosterol content in GL-AmB-treated parasites was ∼2-fold reduced compared with AmB-treated parasites. Moreover, GL-AmB was significantly less cytotoxic and hemolytic than AmB (P<0.01).Conclusion: GNP-based delivery of AmB can be a better, cheaper, and safer alternative than available AmB formulations.Keywords: gold nanoparticle, amphotericin B, antileishmanial, macrophage uptake, ergosterol, immunostimulatorKumar PShivam PMandal SPrasanna PKumar SPrasad SRKumar ADas PAli VSingh SKMandal DDove Medical Pressarticlegold nanoparticleAmphotericin Bantileishmanialmacrophage uptakeergosterolimmunostimulatorMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol Volume 14, Pp 6073-6101 (2019) |
| institution |
DOAJ |
| collection |
DOAJ |
| language |
EN |
| topic |
gold nanoparticle Amphotericin B antileishmanial macrophage uptake ergosterol immunostimulator Medicine (General) R5-920 |
| spellingShingle |
gold nanoparticle Amphotericin B antileishmanial macrophage uptake ergosterol immunostimulator Medicine (General) R5-920 Kumar P Shivam P Mandal S Prasanna P Kumar S Prasad SR Kumar A Das P Ali V Singh SK Mandal D Synthesis, characterization, and mechanistic studies of a gold nanoparticle–amphotericin B covalent conjugate with enhanced antileishmanial efficacy and reduced cytotoxicity |
| description |
Prakash Kumar,1 Pushkar Shivam,*,2 Saptarshi Mandal,*,3 Pragya Prasanna,1 Saurabh Kumar,1 Surendra Rajit Prasad,1 Ashish Kumar,4 Prolay Das,3 Vahab Ali,5 Shubhankar Kumar Singh,2 Debabrata Mandal11Department of Biotechnology, National Institute of Pharmaceutical Education and Research (NIPER) – Hajipur, Vaishali, India; 2Department of Microbiology, Rajendra Memorial Research Institute of Medical Sciences, Patna, India; 3Department of Chemistry, Indian Institute of Technology Patna, Patna, India; 4Department of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences, Patna, India; 5Department of Biochemistry, Rajendra Memorial Research Institute of Medical Sciences, Patna, India*These authors contributed equally to this workBackground: Amphotericin B (AmB) as a liposomal formulation of AmBisome is the first line of treatment for the disease, visceral leishmaniasis, caused by the parasite Leishmania donovani. However, nephrotoxicity is very common due to poor water solubility and aggregation of AmB. This study aimed to develop a water-soluble covalent conjugate of gold nanoparticle (GNP) with AmB for improved antileishmanial efficacy and reduced cytotoxicity.Methods: Citrate-reduced GNPs (∼39 nm) were functionalized with lipoic acid (LA), and the product GNP-LA (GL ∼46 nm) was covalently conjugated with AmB using carboxyl-to-amine coupling chemistry to produce GNP-LA-AmB (GL-AmB ∼48 nm). The nanoparticles were characterized by dynamic light scattering, transmission electron microscopy (TEM), and spectroscopic (ultraviolet–visible and infrared) methods. Experiments on AmB uptake of macrophages, ergosterol depletion of drug-treated parasites, cytokine ELISA, fluorescence anisotropy, flow cytometry, and gene expression studies established efficacy of GL-AmB over standard AmB.Results: Infrared spectroscopy confirmed the presence of a covalent amide bond in the conjugate. TEM images showed uniform size with smooth surfaces of GL-AmB nanoparticles. Efficiency of AmB conjugation was ∼78%. Incubation in serum for 72 h showed <7% AmB release, indicating high stability of conjugate GL-AmB. GL-AmB with AmB equivalents showed ∼5-fold enhanced antileishmanial activity compared with AmB against parasite-infected macrophages ex vivo. Macrophages treated with GL-AmB showed increased immunostimulatory Th1 (IL-12 and interferon-γ) response compared with standard AmB. In parallel, AmB uptake was ∼5.5 and ∼3.7-fold higher for GL-AmB-treated (P<0.001) macrophages within 1 and 2 h of treatment, respectively. The ergosterol content in GL-AmB-treated parasites was ∼2-fold reduced compared with AmB-treated parasites. Moreover, GL-AmB was significantly less cytotoxic and hemolytic than AmB (P<0.01).Conclusion: GNP-based delivery of AmB can be a better, cheaper, and safer alternative than available AmB formulations.Keywords: gold nanoparticle, amphotericin B, antileishmanial, macrophage uptake, ergosterol, immunostimulator |
| format |
article |
| author |
Kumar P Shivam P Mandal S Prasanna P Kumar S Prasad SR Kumar A Das P Ali V Singh SK Mandal D |
| author_facet |
Kumar P Shivam P Mandal S Prasanna P Kumar S Prasad SR Kumar A Das P Ali V Singh SK Mandal D |
| author_sort |
Kumar P |
| title |
Synthesis, characterization, and mechanistic studies of a gold nanoparticle–amphotericin B covalent conjugate with enhanced antileishmanial efficacy and reduced cytotoxicity |
| title_short |
Synthesis, characterization, and mechanistic studies of a gold nanoparticle–amphotericin B covalent conjugate with enhanced antileishmanial efficacy and reduced cytotoxicity |
| title_full |
Synthesis, characterization, and mechanistic studies of a gold nanoparticle–amphotericin B covalent conjugate with enhanced antileishmanial efficacy and reduced cytotoxicity |
| title_fullStr |
Synthesis, characterization, and mechanistic studies of a gold nanoparticle–amphotericin B covalent conjugate with enhanced antileishmanial efficacy and reduced cytotoxicity |
| title_full_unstemmed |
Synthesis, characterization, and mechanistic studies of a gold nanoparticle–amphotericin B covalent conjugate with enhanced antileishmanial efficacy and reduced cytotoxicity |
| title_sort |
synthesis, characterization, and mechanistic studies of a gold nanoparticle–amphotericin b covalent conjugate with enhanced antileishmanial efficacy and reduced cytotoxicity |
| publisher |
Dove Medical Press |
| publishDate |
2019 |
| url |
https://doaj.org/article/303109f71b1a4ab28a430db566d3a540 |
| work_keys_str_mv |
AT kumarp synthesischaracterizationandmechanisticstudiesofagoldnanoparticlendashamphotericinbcovalentconjugatewithenhancedantileishmanialefficacyandreducedcytotoxicity AT shivamp synthesischaracterizationandmechanisticstudiesofagoldnanoparticlendashamphotericinbcovalentconjugatewithenhancedantileishmanialefficacyandreducedcytotoxicity AT mandals synthesischaracterizationandmechanisticstudiesofagoldnanoparticlendashamphotericinbcovalentconjugatewithenhancedantileishmanialefficacyandreducedcytotoxicity AT prasannap synthesischaracterizationandmechanisticstudiesofagoldnanoparticlendashamphotericinbcovalentconjugatewithenhancedantileishmanialefficacyandreducedcytotoxicity AT kumars synthesischaracterizationandmechanisticstudiesofagoldnanoparticlendashamphotericinbcovalentconjugatewithenhancedantileishmanialefficacyandreducedcytotoxicity AT prasadsr synthesischaracterizationandmechanisticstudiesofagoldnanoparticlendashamphotericinbcovalentconjugatewithenhancedantileishmanialefficacyandreducedcytotoxicity AT kumara synthesischaracterizationandmechanisticstudiesofagoldnanoparticlendashamphotericinbcovalentconjugatewithenhancedantileishmanialefficacyandreducedcytotoxicity AT dasp synthesischaracterizationandmechanisticstudiesofagoldnanoparticlendashamphotericinbcovalentconjugatewithenhancedantileishmanialefficacyandreducedcytotoxicity AT aliv synthesischaracterizationandmechanisticstudiesofagoldnanoparticlendashamphotericinbcovalentconjugatewithenhancedantileishmanialefficacyandreducedcytotoxicity AT singhsk synthesischaracterizationandmechanisticstudiesofagoldnanoparticlendashamphotericinbcovalentconjugatewithenhancedantileishmanialefficacyandreducedcytotoxicity AT mandald synthesischaracterizationandmechanisticstudiesofagoldnanoparticlendashamphotericinbcovalentconjugatewithenhancedantileishmanialefficacyandreducedcytotoxicity |
| _version_ |
1718394798355775488 |