Stem cell tumor targeting system with internal nano-prodrug and preparation method thereof
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A technology of tumor targeting and stem cells, which is applied in the field of biomedicine and nanomedicine, can solve the problems that chemotherapy drugs are difficult to guarantee the time requirement, reduce the effect of targeting and anti-tumor, and lose the specific targeting effect, so as to prolong the survival period of animals , enhance the anti-tumor effect, the method is simple and easy to operate
Inactive Publication Date: 2016-04-06
FUDAN UNIV
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However, it is difficult for stem cells to load small-molecule chemotherapeutic drugs to achieve acid-sensitive release of drugs, and the released small-molecule chemotherapeutic drugs still have the problem of being pumped out of the tumor parenchyma, losing their specific targeting effect; premature excretion of chemotherapeutic drugs is also difficult. Guarantee the time required for stem cells to migrate to the tumor tissue within 2 to 4 days, thus affecting the effective accumulation of drugs in the tumor and reducing the targeting and anti-tumor effects
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Embodiment 1
[0072] Example 1: Toxicity investigation of the original drug and nano-prodrug on MSCs:
[0073] Take the MSCs cells in the logarithmic growth phase, and take 10 4 Each well was inoculated in a 96-well plate, 100 μL per well, and after culturing at 37°C for 24 hours, different concentrations of DOX, PPCD and RGD-PPCD (DOXequiv.) were added, and the culture was continued for 24 hours, and then 0.2 mL containing 0.5 mg / For the serum-free culture solution of mLMTT, continue to incubate for 2 hours, then absorb the culture solution, add 0.1mL DMSO, measure the absorbance value (λ=570nm) with a microplate reader after the dissolution is uniform, and calculate the inhibition of cell viability.
Embodiment 2
[0074] Embodiment 2: the investigation of the time frame of MSCs toxicity by former drug and nano-prodrug:
[0075] With the test method of Example 1, the drug concentrations of DOX and PPCD / RGD-PPCD were fixed, and the incubation time was changed to 36h, 48h, and 60h to obtain the inhibition of MSCs cell viability by drugs at different incubation times.
Embodiment 3
[0076] Embodiment 3: MSCs—(RGD-PPCD) n Investigation of the best incubation time:
[0077] MSCs cells in the logarithmic growth phase were taken and co-incubated with DOX, PPCD and RGD-PPCD in culture medium, the drug concentration was 50 μg / mL (DOXequiv.), and incubated in a 37°C incubator for 0h, 1h, 2h, 4h , 8h, 24h, 48h, washed and digested with PBS (pH7.4) at 4°C, and measured the average fluorescence intensity with a flow cytometer (Ex488nm; Em550nm).
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Abstract
The present invention belongs to the technical fields of biomedical medicine and nano-medicine, and relates to a stem cell tumor targeting system with internal nano prodrug and a preparation method thereof. The method uses stem cells as cell carriers to prepare an anti-tumor targeting system with general formula of Stem cells-(RGD-PPCD)n through endocytosis and inner load of nano prodrug. The nano prodrug has the characteristics of targeting, sustained release and acid sensitivity release; and the stem cell carrier can remotely target primary lesion and metastasis of tumor. In vitro experimental results show that the stem cells after drug load can retain the proliferation capacity and tumor migration characteristics; and the constructed system is stable to ensure that the drug stays in the form of nano-prodrug for several days in the cells, and can slowly release the drug. In vivo experimental results show that the constructed stem cell targeting system can significantly prolong the survival time of tumor-bearing animals and can maintain normal neurobehavioral characteristics of tumor-bearing animals in comparison with the original drug and nano prodrugs, so as to play the efficient, secure and long-lasting anti-tumor effect.
Description
technical field [0001] The invention belongs to the technical field of biomedicine and nanomedicine, and relates to a stem cell tumor targeting system loaded with nanometer prodrug and a preparation method thereof. Background technique [0002] According to statistics, at present, malignant tumor is still a serious disease that threatens human health, and its mortality rate (37.0%) is second only to heart disease, ranking second in the cause of death. The main reason is that the tumor infiltrates and grows into the surrounding normal tissues in the early stage of the disease to form satellite lesions. After surgical resection, it is easy to relapse and the degree of malignancy increases, and it is resistant to radiotherapy and chemotherapy. Serious injury, etc., so the survival period of patients with malignant tumors has not been effectively extended. The emergence of nanotechnology provides an opportunity for the effective delivery of drugs to tumors, but there are still ...
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