Study On Insulin-loaded Sanguis Draxonis Nanoparticles For Oral Administration

Diabetes is one of the major diseases that threat to human health,and the most effective drug to treat Typeâ… diabetes is insulin(INS).The clinical use of INS injection and administration for all life bring great suffering and inconvenience for patients.Therefore,we have developed the oral insulin nano-preparations——Insulin-loaded Sanguis Draxonis nanoparticles(ISDN).As one of traditional Chinese medicine,Sanguis Draxonis(SD)can remove blood stasis,swell pain and bleed blood;it can also be used to treat diabetic foot. Another study shows that SD has certain hypolipidemic and hypoglycemic effects,so it can be the adjuvant treatment of diabetes.In this paper,we chose SD as wall material,using precipitation to parcel INS,and made them into ISDN.We then made ISDN into oral powder by addingβ-cyclodextrin(β-CD)in the solution. Preparation and its parameters were studied and the optimal preparation process was determined.RP-HPLC method was developed for the determination of insulin encapsulated in ISDN.Oral hypoglycemic effect of ISDN was also studied.To determine the drug content in nanoparticles,a RP-HPLC method was developed.The ISDN separation was performed on a Phenomenex C₁₈(4.6mm×250 mm,5μm)using a mobile phase consisting of a solution of 0.1 mol·L^-1trifuroacetic acid(pH 3.0)and acetonitrile at 30℃,with the flow rate at 1.0 mL·min^-1and the detection wavelength at 210 nm.The results showed that mass concentration(X)and peak area(Y)had linearity from 5μg·mL^-1to 200μg·mL^-1and the linear equation was Y=1.730×10⁴X-6.002×10⁴ with correlation coefficient r=0.9999.The average recovery was 99.2%,and the average intra-day and inter-day precision were 0.4 and 0.7.Transmission electron microscope(TEM)and scanning electron microscopy (SEM)were utilized to observe the surface characteristics,particle size analyzer was employed to determine the average diameter and its distribution width,and HPLC was developed to determine INS content.The results showed that the ISDN were sphere-like in shape,with the average size was 114±13nm.The average encapsulation rate of the ISDN was 63.6±5.4%,and the average loading capacity was 9.3±0.2 IU·mg^-1.In vitro release study shows that both ISDN and ISDN-β-CD had slow-release effect.After various doses of ISDN-β-CD were intragastrically given to diabetic mice, significant decrease of glucose level was achieved in the 50,70 and 90IU·kg^-1groups. Glucose level was not modified significantly in 30 IU·kg^-1groups.Compared with subcutaneous insulin,oral ISDN-β-CD had long-term hypoglycemic effect.