Beta-casein and its complexes with chitosan as nanovehicles for delivery of a platinum

anticancer drug Syed Zaigham Abbas Kazmi

Ph.D Biochemistry08-arid-1112

Nanotechnology

Design, Synthesis and application of the materials and devices whose size and shape have been engineered at the nanoscale

NanoparticleParticles atleast one dimension smaller than one micron and potentially as small as atomic and molecular length scales

They may be• Amorphous

or• Crystalline

Act as carrier for liquid droplet or gases and can include metal atoms and large molecules such as proteins

Nanoparticles and tumor targetingNanomedicines have enormous potential to improve the precision of cancer

therapy

They have the ability to encapsulate the therapeutic drug and target them to

the tumor cells.

Platinum based anticancer drugsThese are the co-ordinate complexes of platinum

Examples include Cis platin Carboplatin Triplatin

Platinum complexes cause cross linking of the DNA which inhibits the DNA repair and synthesis

Beta Casein and Chitosan as nanovehicle for the drug delivery

A new drug delivery system comprising a novel platinum complex (bipyridine morpho-line dithiocarbamate Pt(II) nitrate) within nanoparticles composed of Beta-casein and chitosan.

The influence of pH on the formation of a colloidally-stable nanocarrier system composed of Pt complex-loaded Beta-CN and chitosan nanoparticles was investigated using

UV–vis spectrometry

Dynamic light scattering (DLS) scanning electron microscopy (SEM)

There is considerable interest in the use of natural biopolymers, such as proteins and polysaccharides, to build particular delivery systems

Nanoparticle formation of proteins and carbohydrates based on the principle of attractive and repulsive forces

Beta-Casein• one of the most abundant proteins in bovine milk and having the

properties of

Self organization Micelles formation

• Beta-CN micelles may be efficient as drug vehicles targeting neo-plastic cells.

• Recent reports suggest that Beta-CN nanoparticles can be used for the entrapment and oral delivery of anti-neoplastic agents

one of the problems of using Beta-CN nanoparticles as potential nanocarriers is that drug molecules may interact with their surfaces

So, keeping in view this problem, Chitosan has been used as a secondary coating for the controlled and targeted release of the drug

ChitosanA cationic polysacchride with increasing number of pharmaceutical and

biomedical applications

Beneficial biological properties include Biocompatibility Biodegradibility Non toxicity Low immunogenecity

• chitosan can interact with proteins especially with whole casein forming either soluble or insoluble complexes depending on the pH

Prepration of platinum complex and its 2 mM stock soln in

distilled water

Prepration of 0.5mg/ml stock soln of Beta casein in 0.1 M

sodium phosphate buffer

Prepration of 0.05% w/v in 0.01% acetate buffer

Pt complex and Beta casein solution in 3:1

Addition of Chitosan stock soln to drug-Beta casein soln

Titartion to achieve the desired pH

DLS and Zeta potential studyThe average size and size distribution of the drug-loaded NPs were

determined using DLS/zeta potential analysis

Particle size distribution was studied by DLS at 25◦C using 0.89 cp for the viscosity of the medium, a fixed angle 90◦for the avalanche photo diode (APD) detector and the wavelength of 678 nm for the 90 mW laser

The zeta potential of nanoparticles was measured using a zeta potential analyzer at 25◦C

The samples were diluted with distilled water and placed in the electrophoretic cell for zeta potential measurements

Scanning electron microscopy (SEM)

pH Mean diameter (Polydispersity) Zeta potential (mV)

3 – Drug aggregation –

5.7 250 ± 5 0.11 ± 0.01 +30 ± 0.6

6.2 270 ± 4 0.17 ± 0.01 +29 ± 1

7 – Precipitation –

CytotoxicityThe cytotoxicity of the drug was studied by using MTT assay

The human colon tumor cell line (HCT-116) was selected to assay the in vitro cytotoxicity of the new synthesized Pt(II) complex, either free or encapsulated in CS– Beta-CN micelles

Optical density of both treated and control cells were taken and the cell viability was calculated by the formula

Cell viability (%) =Atreated/Acontrol × 100

ConclusionNanoparticles consist of CS-Beta-casein loaded with bypyridine

morpholine dithiocarbomate are potentially useful as a vehicle for cancer therapy

Nanocomplex were stable and soluble at pH 5.7 and 6.2 according to SEM, DLS and Zeta potential analysis

Drug efficacy of the Platinum complex was improved by encapsulating in NPs

This newly designed nanocarrier could be a promising candidate for clinical trials for the treatment of different carcinomas

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