Anticancer Activity of Cyclophosphamide Nanoparticles against Ehrlich Ascites Carcinoma Cells Bearing Swiss Albino Mice

Drug delivery systems such as nanoparticles are used to improve the pharmacological and therapeutic properties of anticancer drugs. In this study, a biodegradable polymer chitosan (CS) was used for delivery of cyclophosphamide (CP). Cyclophosphamide loaded chitosan nanoparticles (CP-CS-NPs) were prepared by ion cross-linking to form a uniform particle size of 186.53 ±1.865 nm with 78±2.67 % entrapment efficiency. The release study data at pH 7.4 showed biphasic release where the first rapid phase of release lasted for 6 hr, followed by a slow diffusion order rate of release for the next 24 hr. Anticancer activity of these nanoparticles was investigated in-vivo in Ehrlich ascites carcinoma (EAC) tumour. 2 ×10 6 EAC cells were implanted intraperitoneally (i.p., ascitic tumour) in swiss albino mice. This was assigned as day ‘0’. After 4 days of incubation, mice with developed ascites tumour were administered orally with nanoparticles at two doses i.e.7.5 mg/kg/d and 10 mg/kg/d for 14 days. On 18 th day, animals were sacrificed and blood was collected for estimation of haematological parameters. Ascites fluid was collected for the estimation of tumour volume, cell viability, angiogenic, metastatic and antioxidant parameters. CP-CS-NPs significantly increased total RBCs at a higher dose of 10 mg/kg/d whereas Hb % was increased significantly at both the doses. WBCs were decreased significantly when compared to EAC control group. Furthermore, CP-CS-NPs significantly decreased the tumour volume and cell viability indicating cytotoxic property. Decreased VEGF level and TNF-αand increased IL-12 expression indicated anti-angiogenic potential of CPnanoparticles. These were also able to increase the antioxidant concentration significantly. All results were dose-dependent when compared to EAC control group. The result suggests that CP-CS-NPs elicit anti-carcinogenic activity and might be a potential candidate for developing multifunctional anti-cancer agent. S.S.Agrawal* and Pallavi Sharma 1 DPSR University, New Delhi, India. Submission: 7June 2017 Accepted: 12 June 2017 Published: 25 June 2017 S.S.Agrawal* and Pallavi Sharma 1 DPSR University, New Delhi, India. Submission: 2June 2017 Accepted: 10 June 2017 Published: 25 June 2017 www.ijppr.humanjournals.com Citation: S.S.Agrawal et al. Ijppr.Human, 2017; Vol. 9 (3): 244-265. 245 INTRODUCTION Cancer is one of the leading causes of death around the globe. Around 70 percent of the world’s death occurs in Asia, Africa and Central and South America due to cancer. 1 22 million new cancer cases will emerge within the next two decades. 2 Various approaches have been developed in order to curb the havoc created by cancer worldwide. Current cancer treatment regime includes combination of surgery, radiotherapy and chemotherapy. However, treatment poses many limitations of its own. Surgery leads to removal of sizeable tumour or the entire organ. Radiotherapy disrupts the genetic material of the cells surrounding the treatment area, making impossible for healthy cells to grow. Chemotherapeutics killsor slows the growth of healthy cells.Also, occurrence of multidrug resistance (MDR) due to pglycoprotein action is one of the major threat to treatment plan. 3 Thus novel drug delivery systems (NDDS) such as nanoparticles, liposomes, microspheres etc. have been formulated to circumvent such threats by evading or neutralising p-glycoprotein mediated efflux mechanism. 4 NDDS offers a sustained drug release, thereby reducing frequency of dosing and side effects. 5 Cyclophosphamide is a cytotoxic agent often used to treatretinoblastoma, rhabdomyosarcoma, lymphomas, leukemias, multiple myeloma, neuroblastoma, breast, testicular, endometrial, ovarian, and lung cancers. Traditional dosage forms are encountered with problems like high toxicity and drug leakage before reaching tumours which can be curbed by targeted drug delivery system. 6 However, in this study, we focused on the therapeutic potential of cyclophosphamide when given as chitosan-nanoparticulate dosage form. Nanoparticles are made from different biodegradable and non-biodegradable materials, with dimensions generally less than 500 nm. Nanocarriers used for treatment of a disease have to be biocompatible and nontoxic. 7 Recently, chitosan based nanoparticleshave played a promising role in delivery of cancer chemotherapeutics since chitosan has favourable properties such as mucoadhesivity, biodegradability, low toxicity and notable cell membrane permeability. 8 Commercially, chitosan is formed by deacetylation of chitin. Chitin is a naturally occurring polysaccharide found in exoskeleton of crustaceans such as crabs. 9 Chitosan promotes cross-linkage with various cross-linking agents, such as glutaraldehyde, sodium tripolyphosphate (TPP), genipin etc.Several techniques such as microemulsion,solventevaporation, ionic gelation,etc.have been developed to prepare chitosan nanoparticles.However, for pharmaceutical applications, physical cross-linking utilised in ionic gelation process is more desirable since the cross-linking is reversible and therefore avoid potential toxicity of the reagents. 10 Experimental tumours are extensively www.ijppr.humanjournals.com Citation: S.S.Agrawal et al. Ijppr.Human, 2017; Vol. 9 (3): 244-265. 246 used as models to study anticancer activity. In the current study, Ehrlich ascites carcinoma mouse model has been selected due to resemblance of EAC tumours with human tumours. These undifferentiated tumours have a rapid growth rate and are most sensitive to chemotherapy. MATERIALS AND METHODS