Construction And Bioanalytical Application Of Smart Responsive Optical Probes

Among the analysis techniques,optical analysis is widely used in biochemical detection,disease diagnosis,and drug screening due to its high sensitivity and non-invasiveness.However,with the rapid development of bioscience,traditional optical probes cannot satisfy the requirement of modern biological analysis in sensitivity,selectivity,time-resolved imaging,and functional detection.Owing to the development of chemical biology and nanomaterials science,more and more composite nanomaterials are designed for optical inspection.Among them,the functional modification of the optical nanoprobe can respond to the detection environment characteristics(such as heat,p H,redox,and enzyme)or external regulation(such as light,magnetism,and ultrasound),and achieving flexibility,selection,and intelligent biochemical analysis.In this paper,the luminescence form and wavelength of the probe are optimized for improving the sensitivity.For achieving the selective detection,a series of optical probes based on enzyme-specific catalysis are designed.In order to achieve time-resolved detection of specific events,nanoprobes that can flexibly respond to near-infrared light are prepared.In order to realize the diagnosis and treatment of diseases at the same time,an integrated diagnosis and treatment nanosystem is constructed.The details as follows:(1)For improving the selectivity and sensitivity of optical probes,we designed a novel chemiluminescence probe for biomarker recognition by simulating the two-step metabolic process of H₂O₂ in the peroxisome.The selective detection of analytes was achieved since the specific catalysis of oxidase.Owing to the chemiluminescence form,the probe can sensitively detect the H₂O₂,glucose,lactic acid,uric acid,ethanol,and other biomarkers without background interference.Taking glucose detection as an example,we achieved the chemiluminescence detection of intracellular glucose for the first time.Besides,we successfully established a new method for cell screening of insulin sensitizers.Thus,this peroxisome-inspired nanoprobe holds great promise in the general diagnosis of metabolic diseases and drug discovery.(2)In order to endow small molecule fluorescent probes with universal time-resolved detection capabilities,we prepared two nanoprobes that intelligently respond to near-infrared light for achieving time-resolved monitoring of complex cellular processes and alkaline phosphatase activity in living animals.Firstly,we synthesized an azobenzene derivatives doped lipid membrane upconversion nanoprobe.Through adjusting the power density of the near-infrared light,the upconversion nanocore can achieve two different types of light emissions,and indirectly control the conformational changes of the azobenzene molecules for achieving single-wavelength near-infrared light regulated activation-inactivation of nanoprobes.A reduced nicotinamide adenine dinucleotide nanoprobe was constructed as the model to demonstrate precise and time-resolved monitoring of intracellular processes including cancerous glycolysis and ligand induced enzymatic processes.Secondly,in order to monitor the activity of alkaline phosphatase in different tissues and biological processes with time resolution.We synthesized a novel fluorescent probe based on dichloro-substituted dicyanomethylene-4H-chromene derivatives.The fluorescent probe has a good fluorescence efficiency and stability in the biological p H range,and it was successfully used for the detection of alkaline phosphatase activity in an acidic tumor environment.By further loading the the small molecule fluorescent probe into a near-infrared light-responsive nanocontainer,the time-resolved and long-term imaging of alkaline phosphatase activity were facilely achieved.In addition,we successfully used the nanoprobe to monitor the variation of alkaline phosphatase activity in the mouse model of drug-induced acute liver injury.(3)Based on the successful study of the selectivity,sensitivity,and time-resolved imaging of optical probes,we further developed a dual-near-infrared light orthogonally responded nano-platform for?-glucuronidase activity directed time-resolved nanotheranostic of cancer.To in vivo detect the?-glucuronidase activity,a red-NIR fluorescence probe with deep tissue penetrability was synthesized for the first time.Subsequently,a dual NIR light orthogonally responsive nanotheranostic platform was achieved by using the molecular probe and photodynamic therapy as diagnostic and therapeutic agents,respectively.After independently verification of 980 nm laser remote-controlled?-glucuronidase detection and 808 nm laser regulated photodynamic therapy in vitro.The?-glucuronidase level in various cancer cell lines xenograft tumor models was successfully investigated.The further application of nanotheranostic platform in?-glucuronidase activity-directed specifically labeling and personalized photodynamic therapy of cancer was achieved with orthogonal dual NIR light control.