Study On Biological Characterization, Pharmacokinetics And Efficacy Of Anticancer Drug As Liver-target Nanoparticles

Nano and microparticle carriers have important potential applications for convey a sufficient dose of drugs to the diseased lesions. Polymeric nanoparticles offer a suitable means for delivering various therapeutic agents by either localized or targeted delivery to the tissue of interest. The research in this area is being carried out all over the world at a great pace. The purpose of this paper is to prepare liver-targeting MMC nanoparticles.1. The MMC-PBCA-NP were prepared by the emulsion polymerization method. A rotatable central composite design for three independent variables at three values each was employed. The property of the MMC-PBCA-NP was Dn= (104.1 ± 10.14) nm, EE= (86.56 + 4.20) %, DL= (7.4 + 0.32) %, Q = 0.17 ±0.024.2. A HPLC method was developed to determine mitomycin content in MMC-PBCA-NP. The analytical column was C₁8 (4.6mm × 150mm, 5μm) ;The mobile phase consisted of mixed phosphate buffer-acetonitrile (85:15) and flow rate of 1ml . min^-1. The detection was carried out with UV detector and the detection wavelength of 365nm was specified. Under this condition ,there was agood linear relationship in the range of 5 - 250[ig-mL¹for mitomycin, the correlation coefficient is 0.9996 .The average recovery(n=6) was 98.68%. RSD was 1.52%.3. The biocompatibility by implanting PBCA-NP into muscle and skin of rabbits and in vitro cultured with the L-02 human liver cells. We found the material had no toxicity, no hemolytic activities, and no irritation to muscle or skin. All results indicated that the PBCA-NP had good biocompatibility.4. A comparative study was carried out aimed at evaluating the acute toxicity of MMC and MMC-PBCA-NP by ip. mice. The LD50 of the MMC and MMC-PBCA-NP of mounse was 13.6, 54.3mg ? kg"1 respectively.5. KM mice were injected intravenously via tail veins with MMC and MMC-PBCA-NP in 0.9% of NaCl, pH7.4-7.5(saline) at dose levels of lmg/kg(expresses in MMC content in free and complexed MMC per kg body weight of the mice). At 5,15,30,40,60,180,360,720,1440min after the injections, the mice were sacrificed by cervical dislocating. The liver, kidney, spleen, lung, heart in the mice were immediately removed and washed by lOmM Na2HPC>4 buffer and were then homogenized. For the determination of MMC in different tissue, 3.0ml of ethyl acetate was added as an extracting solvent was added to 1.0ml of tissue homogenates, followed by shaking on a vortex mixer for lmin and centrifugating at 1200xg for 15min. After centrfugation, MMC in the lower layer was determined by HPLC. The concentration-time curves of MMC and MMC-PBCA-NP were fitted to a no-compartment model. The Tl/2 of MMC-PBCA-NP was 2.93-fold higher than MMC. The AUC of serum of MMC-PBCA-NP was 3.29-fold higher than MMC. The tissue concentration of MMC-PBCA-NP in heart, kidney and lung was lower than those of MMC. MMC-PBCA-NP significantly alters its pharmacokinetics and biodistribution inserum and tissues compared to MMC .6. Using rabbits bearing VX2 cells, experiments were performed to investigated the antitumor activity of MMC-PBCA-NP and its adberse effects on the kidney , heart and spleen of the host comparing with those of MMC. Suspension of 4xlO6 of VX2 cells were injected Rabbits with a VX2 tumour implanted into the hindlimb were obtained from Sun Yat-sen university and successive transplantation into the liver of rabbits. Viable tumour tissue taken from the VX2 tumour was minced into 1 * 1 x lmm fragments. Fragments were promptly implanted into the subcapsular parenchyma of the left mediallobe of the liver. When the hepatic mass of the VX2 tumour was clearly established, usually 7-9 days after implantation, the animal were subjected to experimental treatments. The major and minor axes(mm) of the tumour mass were measured by CT . Following this, MMC, MMC-PBCA-NP or saline was injected by the ear vein. The dose levels used were:0.29 mg ? kg"1 for MMC, 0.09,0.29, 0.92 (MMC content) mg kg"1 for MMC-PBCA-NP. The results showed that the antiproliferating effect of the same dose of MMC-PBCA-NP as free MMC were more activity by comparing the sizes and weights of this VX2 grown in the liver of rabbits. Another group of rabbits bearing VX2 cells were sacrificed on the day after the last injection of the drugs. Hearts and kidneys were removed immediately and sections were prepared and stained by standard histological method. The histological sections of the hearts and kidney were observed under light microscope by different combinations of magnification values of the eye pieces and objective lens of the microscope. Micrographs of the heart and kidney sections under microscope were taken for record and comparison. The results in present study indicate that polybutylcyanoacrylate nanoparticles can be used as a targeted carrier for MMC because MMC-PBCA-NP can exhibit similar anti-proliferative effect as MMC on tumor but reduce the MMC inducedthe toxic side effects in the host.