Study On Preparation Technology Of Annonaceous Acetogenins Nanosuspension

Annonaceous acetogenins(ACGs)have been proven to exhibit significant antitumor activity.However,the poor solubility and strong toxicity limits their further application in clinic.Nanosuspensions(NSps)are very helpful to solve the poor solubility and difficulty in the in vivo drug delivery of this kind drug or drug candidates.As a result,ACGs were prepared into nanosuspensions to improve the solubility,stability and cut down side effect and the intravenous injection can increase the drug accumulation in tumor through EPR effect.In this thesis,ACGs were prepared into nanosuspensions by a micro-precipitation combined with ultrasonic approach,and the particle size of NSps were investigated by with a Nano-ZS Zetasizer.the morphology was measured by TEM(transmission electron microscopy).Meanwhile,the vitro properties of NSps slao were investigated by stable in various physiological media,optimization of lyoprotectants for NSps and so on.cytotoxicity assay was measured by MTT assay.The antitumor effect of NSps investigated by in vivo assay.Meanwhile,the acute toxicity study was performed to verified whether nanosuspension can reduce toxic and side effect of ACGs.Firstly,ACGs were prepared into nanosuspensions.And the resultant P188-stabilized ACGs-encapsulated nanoparticles(P188-ACGs-NSps)had a mean particle size of(115.9±2.08)nm and a polydispersity index(PDI)of(0.195±0.01).P188-ACGs-NSps were stable in various physiological media and the cumulative drug release reached 85%at 144 h.With 0.5%glucose as the best lyoprotectant.In comparison with ACGs solution,the NSps exhibited significantly ehnanced cytotoxicity against 4T1(IC50,1.426 ug/mL vs 3.106 ug/mL)and HeLa(IC50,0.718 ug/mL vs 1.521 ug/mL)cell lines.In vivo antitumor study in 4T1 tumor bearing mice results indicated that the intravenous administration of P188-ACGs-NSps(0.4 mg/kg)showed better antitumor efficacy than 8mg/kg of paclitaxel injections,demonstrating that ACGs is more potent antitumor agent than paclitaxel(63%vs 44%).And the acute toxicity study was performed on 1:5 of P188-ACGs-NSps.The result demonstrated that the LD50 values of the P188-ACGs-NSps were 36.86 mg/kg,which was about 30.7 times of oral dose in this thesis.Secondly,ACGs and salinomycin(SAL)were co-encapsulated into nanosuspensions(SAL-ACGs-NSps).And the resultant that SAL-ACGs-NSps had a mean particle size of(171 ±1.1)nm,were nearly spherical observed by TEM and quite stable in various physiological media during the storage at ambient temperature.MTT assay found that in comparison with ACGs-NSps,the suitable SAL-ACGs-NSps exhibited significantly ehnanced cytotoxicity against 4T1(IC50,0.858 ug/mL vs 0.9068 ug/mL)and HeLa(IC50,0.1978 ug/mL vs 0.7028 ug/mL)cell lines.The in vivo study on 4T1 tumor-bearing mice demonstrated that SAL-ACGs-NSps showed higher in vivo antitumor efficacy than paclitaxel(85%vs 52%).The results revealed that combination of ACGs with SAL could achieve the purpose of efficacy enhancement.In summary,the solubility and bioavailability was successfully improved though the preparation of NSps.Meanwhile,the in vivo toxic and side effect of ACGs was reduced to some extent though nanoencapsulation and combinational use of chemosensitizers significantly ehnanced antitumor efficacy.The research results of this thesis lay the foundation for the clinical application of and provide reference for the development of other potent and highly toxic antitumor agents.