Development and evaluation of various pharmacological activities of sustained release formulation of phytochemical PITC-2, isolated from the tissue cultured medicinal plant Pluchea indica (L.)Less.

Abstract

The major treatment for cancer patients is chemotherapies which have various toxic side effects. To overcome these problems various traditional medicines are used. PITC-2 was isolated from the methanolic root extract of tissue cultured medicinal plant Pluchea indica(L.) Less. PITC-2 is a Thiophen derivative which is [2-(Prop-1-ynyl)-5(5,6-dihydroxyhexa-1,3-diynyl)- thiophene]. First objective of the study is to evaluate the in-vivo antitumor activity of PITC 2 against sarcoma- 180 cancer cell and EAC cell line in Swiss albino mice. The antitumor activity was evaluated by treatment with PITC-2 at different dose for 14 days and 21 days on EAC and Sarcoma-180 mice model. The histopathological and immunohistopathological examination indicates that PITC-2 induces apoptosis and suppresses tumor cell proliferation along with G1 cell cycle arrest through the down–regulation of the intratumoral expression of Bcl-2, cyclic D1 and Ki-67 and thus highlighting anti-proliferative and apoptotic properties against sarcoma-180 solid tumor model. Furthermore, treatment with PITC-2 decreases body weight, tumor weight, tumor volume and increases lifespan and survival time of tumor bearing mice. Another major objective of the study is to formulate PITC-2 in a sustained release formulation and evaluation of the prepared formulation. So here we have performed designing, preparation and characterization of PITC-2 loaded Solid Lipid Nanoparticle for i.v. administration. Solid Lipid Nanoparticles were prepared by emulsion evaporation technique. PITC-2 SLNs formed are smooth spherical particles observed in cryo-FESEM with less than 200nm in size. 52% encapsulation efficacy is found with a stable zeta potential of -35nV. DSC and PXRD studies indicate complete encapsulation of drug within the nanoparticle matrix in amorphous form. The drug release study demonstrated a sustained and prolonged drug release from the SLNs. A comparison on therapeutic effectiveness is presented between PITC-2 SLNs and free phytochemical PITC-2 on EAC cell in Swiss albino mice. Treatment with PITC-2 SLNs decreases tumor volume and increases lifespan of cancer bearing mice in comparison to phytochemical PITC-2. The histopathological examination also indicates that PITC-2 SLNs have promising apoptotic activity on tumor cells. Along with this SLNs show no significant manifestation of toxic symptoms on liver and kidney of mice. Hence, the developed PITC-2 loaded SLNs can be used as drug carrier for sustained and prolonged drug release with improve therapeutic activity and bioavailability.