Fc-FF-RGD composite as well as preparation method and application thereof
A complex and reaction solution technology, applied in the field of Fc-FF-RGD complex and its preparation, to achieve the effect of promoting cell adhesion and proliferation, good application potential, and enhancing anti-tumor effect
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Embodiment 1
[0044] Example 1 Preparation of Fc-FF-RGD Complex
[0045] Include the following steps:
[0046] (1) 1g of chlorotrityl resin, with a particle size of 100-200 mesh and a concentration of 0.3-0.8mmol / g, is added to a solid-phase reactor, 30mL of anhydrous dichloromethane is added, and nitrogen gas is introduced to swell for 30 minutes. , remove dichloromethane, and wash the resin three times with anhydrous DMF30-40mL;
[0047] (2) Fmoc-Asp(OtBu)-OH and DIEA were dissolved in 30-40mL of anhydrous DMF, and the solution was added to a solid-phase reactor, and reacted with nitrogen blowing for 1 hour, and then the reaction reagent was removed, And wash the resin three times with 30-40mL anhydrous DMF;
[0048] (3) Add 20 mL of quenching solvent, which is mixed by volume ratio of DCM, MeOH and DIEA, DCM:MeOH:DIEA=80:15:5, react for 10 minutes under nitrogen blowing and shaking, and remove the reaction liquid , then add 20mL of quenching solvent, react under nitrogen blowing and s...
Embodiment 2
[0066] Example 2 Fc-FF-RGD complex biocompatibility detection
[0067] The biocompatibility of the Fc-FF-RGD complex prepared in Example 1 was detected, and the specific steps were as follows: soak the Fc-FF-RGD self-assembled hydrogel in DMEM medium for 24 hours in a 96-well plate, and then Inoculate 4*10 respectively 4 / mL mouse fibroblasts and human umbilical vein endothelial cells were cultured in a cell culture incubator. Cells cultured for 1, 3, and 5 days were collected for cell viability staining, observed under an inverted fluorescent microscope, and cells were counted at the same time. And the microfilaments and nuclei of the cells on the first day were stained to observe the changes of their morphology. From figure 2 It can be seen that with the increase of culture time, mouse fibroblasts and human umbilical vein endothelial cells can adhere to the Fc-FF-RGD self-assembled hydrogel, and can proliferate, and the morphology of the cells is good. This shows that t...
Embodiment 3
[0068] Example 3 Application of Fc-FF-RGD complex as drug carrier
[0069] When the concentration is 1wt% or greater, Fc-FF-RGD can self-assemble into a hydrogel at pH 7.4, and its microstructure is nanofibers. To prepare nanospheres, the concentration must be reduced. In this example, Fc-FF-RGD with a concentration of 0.1 wt% was used to prepare nanospheres with an average diameter of 40 nm at pH=7.4, as shown in image 3 Shown in A. Adding doxorubicin to it, the nanospheres can wrap the doxorubicin inside during the self-assembly process, and use a 1000Da dialysis bag to remove the unwrapped doxorubicin, found under TEM, Fc-FF-RGD Self-assembled to form nanospheres with larger diameters, such as image 3 As shown in B, the average diameter is about 50nm. In order to ensure that the nanospheres can disintegrate inside the cells to release doxorubicin, the pH value was adjusted to 6 in this embodiment, and no nanospheres were observed under the electron microscope, such as ...
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