# US 2020/0399534 A1 · Photo-Activatable Aggregation-Induced Emission Probe with In-Situ Generation Capability, Preparation Method Therefor and Application Thereof > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2020/0399534 A1 - Document ID: US20200399534A1 - Status: Published application - Kind: A1 - Application number: 16/968,426 - Filed: 2018-10-18 - Published: 2020-12-24 - Earliest claimed: 2018-02-09 - Assignees: South China University of Technology - Applicants: South China University of Technology - Inventors: Ben Zhong TANG; Meng GAO; Shiwu LI - Web page: https://patentgrep.com/patent/US20200399534A1 ## CPC - C07D 277/66: with aromatic rings or ring systems directly attached in position 2 - C09K 11/06: containing organic luminescent materials - G01N 21/6486: {Measuring fluorescence of biological material, e.g. DNA, RNA, cells (G01N21/6428 takes precedence)} - G01N 33/5005: {involving human or animal cells (immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025)} - C07D: HETEROCYCLIC COMPOUNDS (macromolecular compounds C08) - C09K: MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) ## Abstract The present invention, belonging to the technical field of biological imaging, discloses a photo-activatable aggregation-induced emission probe with in-situ generation capability, a preparation method therefor and an application thereof. The preparation method is as follows: A bis(2-(2-hydroxybenzylidene)amino)aryl disulfide compound is reacted with thiol to obtain a photo-activatable aggregation-induced emission probe, which has a structure as shown in formula III. In the present invention, the photo-activatable aggregation-induced emission probe is aggregated into a specific organelle, and a 2-(2-hydroxyphenyl)benzothiazole compound having the aggregation-induced emission property is generated through a photo-oxidation reaction. The photo-activatable aggregation-induced emission probe generated in situ of the present invention can effectively overcome the defects of aggregation-induced quenching of traditional fluorescent dyes, and realize organelle-targeted specific photo-activatable fluorescence imaging in living cells, having such advantages as easy preparation, long-term storage, high photo-activation efficiency, large Stokes shift, and strong ability to enter cells. ## Claims ### Claim 1 A photo-activatable aggregation-induced emission probe with in-situ generation capability, wherein the emission probe has a structure as shown in formula III: wherein in formula III, R and R1 are independently hydrogen, halogen, ester group, cyano group, nitro group, substituted or unsubstituted alkyl, alkoxy, alkylamino group, alkylthio group, aryl, heteroaryl, aryloxy, arylamino group, arylthio group, heteroaryloxy, heteroarylamino group, and heteroarylthio group. ### Claim 2 (depends on 1) The photo-activatable aggregation-induced emission probe with in-situ generation capability according to claim 1, wherein the substituted alkyl means that the hydrogen in the alkyl is replaced by a group formed by a cyclic compound, wherein the cyclic compound is a cyclic compound formed by carbon and hydrogen and one or more of heteroatoms N, S and O. ### Claim 3 A method of preparing a photo-activatable aggregation-induced emission probe with in-situ generation capability, wherein the method comprises obtaining the photo-activatable aggregation-induced emission probe by reacting a compound of formula I with a compound of formula II; wherein: the structure of the photo-activatable aggregation-induced emission probe is as shown in formula III: the compound of formula I is a bis(2-(2-hydroxybenzylidene)amino) aryl disulfide compound, having a structure as shown in the following formula I: the compound of formula II is thiol, having a structure as shown in formula II: HS—R2(I); in formulas I and III, R and R1 are independently hydrogen, halogen, ester group, cyano group, nitro group, substituted or unsubstituted alkyl, alkoxy, alkylamino group, alkylthio group, aryl, heteroaryl, aryloxy, arylamino group, arylthio group, heteroaryloxy, heteroarylamino group, and heteroarylthio group; and R2 is a substituted or unsubstituted alkyl and aryl; the substituted alkyl is hydroxyl substituted, ester-group substituted or carboxyl substituted alkyl; the hydroxyl substituted alkyl is an alkyl on which hydrogen is replaced by a hydroxyl; the ester-group substituted alkyl is an alkyl on which carbon is replaced by an ester group; the carboxyl substituted alkyl is an alkyl on which carbon is replaced by a carboxyl; or the thiol is mercapto-containing amino acid or mercapto-containing polypeptide. ### Claim 4 (depends on 3) The method of preparing the photo-activatable aggregation-induced emission probe with in-situ generation capability according to claim 3, wherein: in formulas I and III, the substituted alkyl means that the hydrogen in the alkyl is replaced by a group formed by a cyclic compound, wherein the cyclic compound is a cyclic compound formed by carbon and hydrogen and one or more of heteroatoms N, S and O; in formulas I and III, the alkyl is a C1-30 alkyl, the alkoxy is a C1-30 alkoxy, the alkylamino group is a C1-30 alkylamino group, and the alkylthio group is a C1-30 alkylthio group; in formulas I and III, the heteroaryl refers to a monocyclic or polycyclic aromatic group with 6-20 carbon atoms; the aryls in the aryloxy, arylamino group and arylthio group are each independently a monocyclic or polycyclic aromatic group with 6-20 carbon atoms; the heteroaryl refers to a monocyclic or polycyclic heteroaromatic group with 1-20 carbon atoms and 1-4 heteroatoms selected from N, S and O; the heteroaryls in the heteroaryloxy, heteroarylamino group, and heteroarylthio group are each independently a monocyclic or polycyclic heteroaromatic group with 1-20 carbon atoms and 1-4 heteroatoms selected from N, S and O, and carbon and heteroatoms can form cyclic groups; and in formula II, R2 is a substituted or unsubstituted C1-30 alkyl. ### Claim 5 (depends on 3) The method of preparing the photo-activatable aggregation-induced emission probe with in-situ generation capability according to claim 3, wherein the method comprises carrying out the reaction in an organic solvent; wherein: the organic solvent is more than one of dimethyl sulfoxide, N,N-dimethylformamide, acetone, acetonitrile, dichloromethane, trichloromethane or tetrahydrofuran; the reaction temperature is 15° C. to 40° C., and the reaction time is 1-30 min; and the molar ratio of the compound of formula I to the compound of formula II is from 1:2 to 1:10. ### Claim 6 (depends on 3) The method of preparing the photo-activatable aggregation-induced emission probe with in-situ generation capability according to claim 3, wherein the compound of formula I is a bis(2-(2-hydroxybenzylidene)amino)aryl disulfide compound, the disulfide compound prepared through: dissolving a compound of formula V in an organic solvent, and adding hydrogen peroxide at 20° C. to 60° C. to generate a compound of formula VI; and reacting the compound of formula VI with a compound of formula VII under reflux in an organic solvent to obtain the compound of formula I; wherein in the compound of formula V the compound of formula VI and the compound of formula VII the definitions of the substituents R and R1 are the same as in the compound of formula I. ### Claim 7 (depends on 1) The method of preparing the photo-activatable aggregation-induced emission probe with in-situ generation capability according to claim 1, wherein the method comprises: reacting the compound of formula V with the compound of formula VII in the dark in an organic solvent and a protective gas to obtain the photo-activatable aggregation-induced emission probe; wherein in the compound of formula V and the compound of formula VII R and R1 are independently hydrogen, halogen, ester group, cyano group, nitro group, substituted or unsubstituted alkyl, alkoxy, alkylamino group, alkylthio group, aryl, heteroaryl, aryloxy, arylamino group, arylthio group, heteroaryloxy, heteroarylamino group, and heteroarylthio group. ### Claim 8 (depends on 1) A method of preparing an aggregation-induced emission compound, wherein the method comprises subjecting the photo-activatable aggregation-induced emission probe with in-situ generation capability, as defined in claim 1, to photo-oxidation to obtain the aggregation-induced emission compound. ### Claim 9 (depends on 1) An application of the photo-activatable aggregation-induced emission probe with in-situ generation capability according to claim 1 in biological imaging. ### Claim 10 (depends on 1) An application of the photo-activatable aggregation-induced emission probe with in-situ generation capability according to claim 1 in organelle specific fluorescence imaging, wherein fluorescence imaging is achieved through photo-activation.