Antitumor Study Of Enzyme Loaded Calcium Phosphate Catalysis With Elemene Extracted From Traditional Chinese Medicine

With the development of enzyme and drug-carrying material,nanodrugs,and nano-catalysis have gradually emerged as nano-catalysis drug therapy.Various diseases such as cancer,cardiovascular disease,and diabetes have been found to be related to the generation of free radicals by nano-catalysis.Many Traditional Chinese Medicines can improve the deficiency of healthy,body imbalances,and blood stasis,dampness coagulation and toxin accumulation by regulating the redox stress state of cells,thus achieving anti-tumor,anti-oxidant,and anti-inflammatory effects.Nano-catalysis drugs have outstanding advantages such as low toxicity,long circulation,and good biocompatibility,but their low biodegradation,low enzyme loading capacity and exogenous problems still need to be addressed.Nanosizing technology of loaded materials can improve the water solubility and bioavailability of the active ingredients of drugs.Therefore,the following research work was carried out in the thesis for the nanofabrication of catalytic drug and their combination with traditional Chinese medicine elemene.1.We investigated the fabrication process and properties of magnesium carbonate,calcium carbonate,and amorphous calcium phosphate as basic nanocarrier materials.Enzyme loaded particle of magnesium carbonate,calcium carbonate,and calcium phosphate were prepared by co-precipitation method.Through the comparison of the morphology and catalytic effect,it was found that amorphous calcium phosphate and glucose oxidase could form homogeneous nanospheres while maintaining high catalytic activity through its unique hydration structure.It showed better biological application prospect than magnesium carbonate and calcium carbonate.2.We explored the load capacity of amorphous calcium phosphate on different drugs and proteins.Among them,the loading of glucose oxidase reached 97.3%,and that of bovine serum protein and fermented cordyceps polysaccharide reached 55.2%and 56.3%,respectively.It is indicated that some proteins and natural drugs can be loaded rapidly by calcium phosphate co-precipitation method,which provides an effective route to enhance the loading capacity of proteins and drugs.3.The particle size of the glucose oxidase nanoparticles was rapidly modulated.Nanoparticles with controllable particle size and high stability can be obtained by nanochannels extrusion method.Particles with precise particle size control capability can be suitable for intracellular application.It has been used as an intracellular glucose detector with high biosecurity without destroying cell structure.It can be used in combination with fluorescent probes not only to distinguish tumor cells with different rates of glucose metabolism,but also to analyze and screen the glucose metabolism status in different cells,providing basic metabolic data for cancer diagnosis and treatment.4.Elemene was combined with cascade catalyzed particles under the theory of“molecular compatibility”to achieve the antitumor purpose in a safer and more effective way.The apoptosis experiment showed that compared with the control group,the combination of the two drugs could increase the early and late apoptosis by 15.57%and 14.08%,exceeding the effects of elemene or particles alone on early and late apoptosis of cells.The membrane was coated on the surface of cascade catalyzed particles using the principle of biomimetic,to realize the highly biocompatible anti-tumor application.The IC₅₀ was 7.83μg/m L by cytotoxicity assay.The results showed that the combination of nanoscale catalytic drug technology and effective components of traditional Chinese medicine can synergically and efficiently exert the anti-tumor effect,and improve the in-vivo delivery efficiency of nanoparticles through surface modification.