Nano-Co-Delivery of Berberine and Anticancer Drug Using PLGA Nanoparticles: Exploration of Better Anticancer Activity and In Vivo Kinetics
Combinatorial approach can be beneficial for cancer treatment with better patient recovery. Co-delivery of natural and synthetic anticancer drug not only valuable to achieve better anticancer effectivity but also to ascertain toxicity. This study was aimed to co-deliver berberine (natural origin) and doxorubicin (synthetic origin) utilizing conjugation/encapsulation strategy through poly (lactic-co-glycolic acid) (PLGA) nanoparticles.
Doxorubicin was efficiently conjugated to PLGA via carbodiimide chemistry and the PLGA-doxorubicin conjugate (PDC) was used for encapsulation of berberine (PDBNP).
Significant anti-proliferative against MDA-MB-231 and T47D breast cancer cell lines were observed with IC50 of 1.94 ± 0.22 and 1.02 ± 0.36 μM, which was significantly better than both the bio-actives (p < 0.05). The ROS study revealed that the PDBNP portrayed the slight increase in the reactive oxygen species (ROS) pattern in MDA-MB-231 cell line in a dose-dependent manner, while in T47D cells, no significant change in ROS was seen. PDBNP exhibits significant alteration (depolarization) in mitochondrial membrane permeability and arrest of cell cycle progression at sub G1 phase while the Annexin V/PI assay followed by confocal microscopy resulted into cell death mode to be because of necrosis against MDA-MB-231 cells. In vivo studies in Sprague Dawley rats revealed almost 14-fold increase in half life and a significant increase in plasma drug concentration.
The overall approach of PLGA based co-delivery of doxorubicin and berberine witnessed synergetic effect and reduced toxicity as evidenced by preliminary toxicity studies.
Key Wordsberberine encapsulation in vivo pharmacokinetics mitochondrial pathway necrosis PLGA-doxorubicin conjugate
Atomic force microscopy
Human Peripheral Blood Mononuclear Cells
Berberine loaded PLGA-doxorubicin nanoparticles
Red blood cells
Reactive oxygen species
Scanning electron microscopy
ACKNOWLEDGMENTS AND DISCLOSURES
The authors would like to acknowledge the financial support received from Department of Science and Technology and University Grants Commission, New Delhi, India to Dr. Umesh Gupta in the form of DST Start up Research Grant (for Young Scientists). The first author (IK) also would like to acknowledge Indian Council of Medical Research (ICMR), New Delhi (Award letter no. 45/12/2018-Nan/BMS) for providing Senior Research Fellowship (SRF). GJ thanks CSIR, New Delhi (Grant no. 05/1051(0011)/2018-EMR-I) for providing SRF. The authors declare no competing financial interest. The authors would also like to acknowledge the Central Instrumentation Laboraory Central University of Punjab for extending facilities to carry out microscopy.
- 9.Nabholtz JM, Falkson C, Campos D, Szanto J, Martin M, Chan S, et al. Docetaxel and doxorubicin compared with doxorubicin and cyclophosphamide as first-line chemotherapy for metastatic breast cancer: results of a randomized, multicenter, phase III trial. J Clin Oncol. 2003;21:968–75.CrossRefGoogle Scholar
- 10.Henderson IC, Berry DA, Demetri GD, Goldstein LJ, Martino S, Ingle JN, et al. Improved outcomes from adding sequential paclitaxel but not from escalating doxorubicin dose in an adjuvant chemotherapy regimen for patients with node-positive primary breast cancer. J Clin Oncol. 2003;21:976–83.CrossRefGoogle Scholar
- 11.Burger H, Foekens JA, Look MP, Meijer-van Gelder ME, Klijn JG, Wiemer EA, et al. RNA expression of breast cancer resistance protein, lung resistance related protein, multi-drug resistance gene-1 in breast cancer: correlation with chemotherapeutic response. Clin Cancer Res. 2003;9:827–36.PubMedGoogle Scholar
- 15.Ebrahimian M, Taghavi S, Ghoreishi M, Sedghi S, Farzad SA, Ramezani M, et al. Evaluation of efficiency of modified polypropylenimine (PPI) with alkyl chains as non-viral vectors used in co-delivery of doxorubicin and TRAILplasmid. AAPS Pharm Sci Tech. 2017;19:31029–36.Google Scholar
- 20.Malvern. ISO13320 2009. Particle Size Analysis - Laser Diffraction Methods, Part 1: General Principles.Google Scholar
- 23.Kumar H, Gothwal A, Khan I, Nakhate KT, Alexander A. Ajazuddin, et al. galactose anchored gelatin nanoparticles for primaquine delivery and improved pharmacokinetics: a biodegradable and safe approach for effective anti-plasmodial activity against P. falciparum 3D7 and in vivo hepatocytes targeting. Mol Pharm. 2017;14:3356–69.CrossRefGoogle Scholar
- 32.Langdon SP. Cancer cell culture, methods and protocols, methods in molecular medicine, Humana Press, 2004.Google Scholar
- 33.Meerloo JV, Kaspers GL, Cloos J. Cell sensitivity assays: the MTT assay. Cancer Cell Culture. 2011:237–45.Google Scholar