Application Of Fluorescent Sensor In The Reserach Of Anti-leukemia Drugs

Leukemia is a malignant disease caused by abnormal proliferation and differentiation of hematopoietic stem cells.At present,the commonly used treatment methods are chemotherapy,immunotherapy and radiotherapy,but their prognosis is poor.Targeted drug delivery can induce drugs to enter the target cells or tumor microenvironment,thus alleviating systemic adverse reactions.However,some anticancer drugs have poor solubility and strong toxicity,which is one of the reasons leading to short prognosis and disease recurrence.Hypermolecular macrocyclic compounds(crown ether,cyclodextrin,calix[n]arene,cucurbit[n]urils)can encapsulate drugs into their own cavities to form molecular capsules,which can improve the solubility of drugs,reduce toxic and side effects and administration time.In recent years,supramolecular fluorescence sensors based on macrocyclic compounds have been widely used in the detection of ions,neutral molecules and biomacromolecules,and have made progress in the field of drug delivery.Cucurbit ring can encapsulate drug guest molecules in hydrophobic cavities through hydrogen bonds,dipoles and other noncovalent actions to form stable cucurbit ring@drug complexes,and release drug molecules in response to changes in competitive guest molecules or microenvironment.However,cucurbit ring itself has no fluorescence characteristics,so it is necessary to introduce fluorescent materials to construct supramolecular fluorescence sensors based on cucurbit ring.Tetrastyrene is a compound with propeller molecular configuration and aggregation induced luminescence.When the compounds aggregate,the intermolecular distance decreases,which leads to the inability of C-C on stilbene to rotate freely,and the intramolecular movement is restricted,thus enhancing the fluorescence.Tetrastyrene and its derivatives have good photostability and no aggregation fluorescence quenching effect of other fluorescent materials,so they have great application prospects in the field of cell imaging and drug delivery.Chapter 1: Firstly,the concept and common mechanism of fluorescence sensor are summarized,and then the application of supramolecular fluorescence sensor based on macrocyclic compounds in ion and molecule detection is introduced.Application progress of cucurbit ring in drug delivery and release and research on fluorescent modification of cucurbit ring.The research and application progress of aggregationinduced luminescence effect and tetrastyrene derivatives are reviewed comprehensively.Finally,the importance of eukaryotic elongation factor 2 kinase in tumor cells is introduced.On this basis,we use the newly synthesized supramolecular fluorescence sensor TPE-2Q[7] of our research group to construct a visual targeted drug delivery system of e EF2 K selective inhibitor NH-125,study the interaction between TPE-2Q[7] and NH-125,evaluate its effect on the anti-leukemia activity of NH-125,and provide a theoretical basis for the possibility of e EF2 K as a target for the treatment of leukemia.The main contents of this paper include the following aspects:Chapter 2: The active drug NH-125 was screened by MTT colorimetric method,and the host-guest interaction between TPE-2Q[7] and NH-125 was studied by ultraviolet absorption spectroscopy and fluorescence spectroscopy.The inclusion ratio of TPE-2Q[7] and NH-125 was 1:2,and the binding constant Ka was 1.38 × 106L?mol-1.Chapter 3: The anti-leukemia activity of different concentrations of NH-125,TPE-2Q[7] and TPE-2Q[7]@NH-125 was detected by MTT colorimetric method and flow cytometry.It was found that(1)The anti-leukemia activity of NH-125 is timeand concentration-dependent,which can significantly induce the late apoptosis of K562 cells(P < 0.05),arrest G2/M phase and significantly reduce mitochondrial membrane potential.(2)TPE-2Q[7] has no significant effect on cell growth,apoptosis,cell cycle and mitochondrial damage.(3)Compared with NH-125 alone,TPE-2Q[7]@NH-125 has no significant change in cell growth inhibition activity,but it can significantly increase the percentage of apoptosis induced by NH-125(P < 0.05),significantly increase the number of cells in G2/M phase(P < 0.01)and significantly reduce mitochondrial membrane potential(P < 0.01)Chapter 4: The effectiveness of TPE-Q[7] carrying NH-125 into K562 fine cells was investigated by Western blot.The results showed that(1)TPE-2Q[7]@NH-125 mainly played a role in cytoplasm.(2)TPE-2Q[7] did not affect the inhibitory effect of NH-125 on e EF2 K.(3)TPE-2Q[7]@NH-125 and NH-125 have the same effect,that is,the fluorescent sensor carrying NH-125 can effectively inhibit e EF2 K in cells,induce phosphorylation of e EF2,and ensure the effectiveness of NH-125 entering leukemia cells.To sum up,TPE2Q[7]@NH-125,a visual targeted drug delivery system based on supramolecular fluorescence sensor,can realize the real-time monitoring of drugs and carry two times of drugs into cytoplasm,increase the percentage of apoptosis induced by drugs,increase the number of cells in G2/M phase of drug block and aggravate the degree of mitochondrial damage.TPE-2Q[7]@NH-125 can enhance the anti-leukemia activity of drugs to some extent.