Anticancer Effects Of Tumor-blood Vessel-targeting Long-circulating Microemulsions Loaded With Oil Of Jobstears Seed

Objective:One of the significant progresses in the research of tumor is establishing the important position of the tumor angiogenesis in tumor development and the inhibition of tumor angiogenesis may be the best means of cancer treatment. At present, the main tumor angiogenesis inhibitors are: chemotherapy drugs, biological products, traditional Chinese medicine and tumor targeting drug delivery systems.In the research field of inhibition of tumor angiogenesis chemotherapy drug, Metronomic Chemotherapy is a more successful approach, and its effectiveness has been confirmed by many domestic and foreign hospitals. The essence of Metronomic Chemotherapy is the use of Metronomic Chemotherapy drugs, frequent delivery, thereby inhibiting tumor angiogenesis newborn. Cut off nutritional source of tumor cells, so as to achieve the purpose of inhibiting tumor growth. Biological treatment of tumors uses biological products mainly, among which VEGF and VEGFR-2 receptor antibodies are most important. VEGF is one of the most important factors for promoting angiogenesis. There are many VEGF receptors and KDR plays an important role in the process of tumor angiogenesis newborn. Thus VEGF antibody and KDR antibody can selectively inhibit tumor angiogenesis newborn. They both have good prospects. VEGF is secreted by the tumor cells and spreads to the tumor vascular endothelial cells by body fluid to excite KDR on the surface of vascular endothelial cell. Promote tumor angiogenesis newborn. Different from VEGF antibody, KDR is expressed by tumor vascular endothelial cells and as it's on the surface of tumor vascular endothelial cell, its antibody not only can effectively inhibit tumor angiogenesis newborn, but also has higher tumor-blood vessel- targeting.Chinese traditional medicine has been playing an important role in the field of tumor therapy. It has been confirmed that many kinds of Chinese medicines or their active ingredients are effective in inhibiting the formation of tumor angiogenesis .In recent years, people have found that the anti-tumor drug—oil of jobstears seeds which has entered clinical application has a strong role in Inhibiting angiogenesis newborn.Tumor targeting drug delivery system is mainly used for the selective delivery of anticancer drugs to tumor cells or tumor angiogenesis. The Long-circulating technology is relatively successful. In recent years, we have developed a long-circulating microemulsion. Polyethylene glycol chain is inlaid on the surface of long-circulating microemulsion in order to prevent microemulsion from being phagocytizing and destroying by reticuloendothelial system. So Microemulsion can long-term circulate in the blood system. Its main advantage is to reduce drug to the normal tissues (such as the heart) distribution and promote drug to certain vascular permeability relatively high lesions (such as tumors) distribution.The main content of this study is to connect KDR antibody to the surface of Microemulsions loaded with oil of jobstears seed, so as to achieve tumor-blood vessel-targeting. Animal experiments show that tumor-blood vessel-targeting long-circulating microemulsion has significantly improved the anti-tumor effect of Oil of Jobstears Seeds. In this experiment tumor-blood vessel-targeting long-circulating microemulsion can't lead to animals'death under the largest dose, but can reduce the animals'food intake rate and body weight. After stopping administrating, food intake rate can be back to normal and body weight can also gradually resume. Analgesic experiments show that tumor-blood vessel-targeting long-circulating microemulsion doesn't lower the analgesic effect of Oil of Jobstears Seeds.Methods:1.After preparing Tumor-blood vessel-targeting Long-circulating Microemulsions Loaded with Oil of Jobstears Seed (KDR-LM), use scanning electron microscopy to observe KDR-LM's shape and diameter through frozen etching technology in order to complete KDR-LM's quality evaluation.2. Through an Acute Toxicity Test to evaluate KDR-LM's acute toxicity test, understanding the animal toxicity and its severity after single drug administration, or in a short period of time several times drug administration, to provide certain information for clinical medication security and monitoring.3. Acute Toxicity Test3.1 Explore dose range: based on experience or documentation to identify animals 100% and 0% of the dose of death, that is, lower-dose and dose limit (Dm,Dn). To non-toxic references drugs generally prepare the largest concentrations of pharmaceutical, and then according to geometric relationships (such as 2:1) dilute into multiple doses. Each group has 4 - 10 mice. Give medicine according to the estimated dose. If all die, lower dose. If all live, increase dose. Thereby define Dm, Dn.3.2 Identify several groups and calculate the dose of each group: Group (G):5-8 Groups. Each group dose: the doses are in accordance with the geometric progression. According to r = (G-1) Dm / Dn, calculate the dose of each group: D1, D2, D3, D4, D5,... Dm. D1=the smallest dose, D2=D1*r, D3=D2*r,... Dm=Dn-1*r. Prepare geometric dilution solution: Mice administration requires the same volume, according to the formula (Cm = Dm / isovolumic injection volume) calculate the concentration3.3 Experiment: Packet and edit numbers: Mice are randomly divided into 4 groups, each with 10, half male and half female. Give drugs: Animals of control group are given intravenous injection slowly of saline according to 1 ml/20g, the other three groups are injected from tail vein for 5 days according to a dose of 1 ml/20g. Mice each day are ensured sufficient feed(500g) and water, The mice food intake rate is calculated according to the following formula : Food intake rate = Daily feed intake of each group animal / (10) [g / (number of animals, days)], at the same time have a measure of the average weight of each group animals. Observe and record mice average body weight and food intake rate, until the10th day after injection.4. Anti-tumor effects experiment4.1 Through the right axillary forelimb vaccinate S-180 entity type sarcoma, and establish a mouse model of sarcoma. The day after vaccinating, divide them into 8 groups according to the principle of minimum weight difference and an equal number of male and female animals. Dosage is high and low.4.2 Evaluate by two programs of administrating Tumor-blood vessel-targeting Long-circulating Microemulsions Loaded with Oil of Jobstears Seed's anticancer effect.Traditional treating method, that is the large and single dose treating method (2 mg/20g animal, 4 mg/20g animal) and small dose chemotherapy (0.25 mg/20g animal 24 hour intervals, the administration 8 times; 0.5 mg/20g animal 48 hours intervals, administration 8 times). At the same time record the changes in tumor volume. Kill these animals after stopping giving medicine 24 hours, weighing weight, anatomizing and dissecting the block, weighing its weight, calculating the rate of inhibition of tumor growth.5. Experiment of AnalgesiaUse hot plate method to measure the mice's pain threshold. According to the pain threshold of each group mice, evaluate analgesic effects of various Oil of Jobstears Seeds. Results:1.Use scanning electron microscopy to observe KDR-LM's shape and internal structure. According to the photos, the shape of KDR-LM is round and its diameter is less than 50 nanometers2.In this experiment the animals which are given the largest dose of Microemulsions Loaded with Oil of Jobstears Seed(M), Long-circulating Microemulsions Loaded with Oil of Jobstears Seed(L-M), and KDR-LM by Intravenous didn't die after administration within 10 days, but compared with the control group , animal body weight decreased significantly and food intake rate was also significantly decreased. However, after stopping administration animal body weight began to be picked up and food intake rate also resumed.3. Through single large dose intravenous administration, compared with M and L-M , KDR- LM anti-tumor effects are significantly improved(P<0.01). Give L-M and KDR-LM and we can have significant anti-tumor effect, but animal body weight doesn't grow or grows slowly.4. Using multiple low-dose regimen, that is, small dose chemotherapy, L-M or KDR-LM have significantly improved the anti-tumor effects of Oil of Jobstears Seed(OJS) (P<0.01). However, the animal weight grows slowly. In addition, the anti-tumor effect of KDR-LM is significantly higher than carried by the L-M (P<0.05).5. Moreover, the use of low-dose regimen, the animals of KDR-LM and L-M treating groups obviously Increase body weight, in close to the control group .Suggest that the use of low-dose treatment, not only ensures the anti-tumor effect, but the quality of life of animals may also not be affected.6. KDR-LM retains the analgesic effect of OJS, at the same time also allows the analgesic effect of OJS to extend.Conclusion:1. LM has increased the anti-tumor effect of OJS.2. KDR-LM further enhances the anti-tumor effect of OJS.3. KDR-LM is more suitable for the small tumor dose treatment. Because it can not only improve the anti-tumor effect of the OJS, but also ensure the quality of life of animals not to be seriously affected.4. KDR-LM can reserve analgesic effect of OJS, at the same time can also extend its analgesia time.