| Brucea javanica oil (BJO) is extracted from the nucleoli of Brucea javanica (L) Merr (Simaroubaceae) which is widely distributed from Southeast Asia to northern Australia Brucea javanica is characterized as an antipyretic and detoxifying plant, and widely used in the treatment of lung, prostate, and gastrointestinal cancer, and has potent antimalarial, anti-inflammatory, and antiviral effects, with low toxicity. But application of BJO is restricted by its poor solubility, and BJO is administered as an emulsion (BJOE) for intravenous use. However, due to the thermodynamic instability of emulsions, BJOE easily delaminates, flocculates, and rancidifies during the storage. Additionally, because BJO is a traditional Chinese medicine of bitter and cold, it is bad for the stomach and intestines. To alleviate its negative effects on the digestive tract and improve its curative effects, it is essential to develop new formulation.CDP was obtained by reaction of β-CD with epichlorohydrin (EPI). And a new water-soluble inclusion complex of Brucea javanica oil (BJO) with β-cyclodextrin polymer (CDP) was prepared by saturated aqueous method.1 Preparation of BJO-CDP① CDP was obtained by reaction of β-CD with epichlorohydrin (EPI) and characterized by SEM,1H NMR, FT-IR and the change of melting range.② BJO-CDP was prepared by saturated aqueous method and characterized by SEM,1H NMR, FT-IR and the change of melting range.③ A HPLC method was established by using Brusatol as the index components and methodological investigation was carried out to evaluate the method. The results of HPLC showed that the peak area of Brusatol had a good linear relationship. The regression equation of Brusatol was Y=1 ×107X+987534, R2=0.999. The specificity, precision and stability all meet the requirement which showed that the HPLC method can be used to determine the content of the inclusion complexes.④Use single factor experiment to filter apposite mass ratio of BJO and CDP, temperature, time and concentration of solvent for orthogonal test.⑤The multiple factors including mass ratio of BJO and CDP, time, temperature and concentration of solvent were investigated in the inclusion process to optimize the preparation conditions of BJO-CDP. The result showed that the optimum preparation conditions for inclusion were established as follows:the mass ratio of BJO and CDP was 1:10, time was 6 min, temperature was 20 ℃ and the amount of ether was 0.5g.2 Quality Inspection①Research the solubility according to the legend items of the solubility determination in China Pharmacopoeia (version 2015) in different media of the lyophilized powder was determined. The BJO-CDP has good solubility in distilled water, artificial gastric juice and artificial intestinal fluid but almost insoluble in methanol, ethanol.②Melting range was also detected. The melting range of -CD was 298～305 ℃, CDP was 271～278℃, BJO-CDP was 267～274℃.③The stability of the BJO-CDP inclusion complexes was detected by putting them under the condition of illumination, high temperature and high humidity. The results showed that BJO-CDP has good stability under illumination and high temperature. The inclusion complexes becomes toughness and agglomeration at high humidity condition which indicated that we should try to keep dry at the procedure of preparation、storage and transport of BJO-CDP.3 Pharmacodynamics of BJO-CDP①The pharmacodynamics of BJO-CDP inclusion complexes was investigated using MMC-7721 cell lines. The commercial emulsion of BJO (BJOE) was used as a reference. The IC50 values obtained in BJO-CDP group and emulsions group were 0.9518 and 1.219mg/ml respectively. Flow cytometry revealed that medium and high dosage of drugs had significant role in inducing apoptosis of 7721 cells②In the experiment that the inhibition effects on mice colon cancer, significant inhibition effects on colon cancer was observed in BJO-CDP groups with middle dose (p<0.05) and high dose (p<0.05), whereas in BJOE groups, significant effect was only observed with high dose (p<0.05). Moreover, the BJO-CDP groups had less weight loss than BJOE groups. These results indicates that BJO-CDP had stronger antitumor activity with a lower toxitity than BJOE.③In the acute toxicity test, the median lethal dose (LD50) of BJOE was 9.78 g/kg. In contrast, all mice treated with inclusion complexes survived even at the highest dosage (15.36 g/kg).In conclusion, the present study suggests that BJO-CDP polymer significantly decreased toxicity of BJO and enhance the antitumor activity. Therefore, β-cyclodextrin plomer (CDP) may be a potential effective delivery vehicle for this poorly soluble antitumor drug. |
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