CN112552507B - Trigger type self-degradation polymer-based near-infrared fluorescent probe, preparation method and application - Google Patents

Trigger type self-degradation polymer-based near-infrared fluorescent probe, preparation method and application Download PDF

Info

Publication number
CN112552507B
CN112552507B CN202011200558.6A CN202011200558A CN112552507B CN 112552507 B CN112552507 B CN 112552507B CN 202011200558 A CN202011200558 A CN 202011200558A CN 112552507 B CN112552507 B CN 112552507B
Authority
CN
China
Prior art keywords
polymer
self
degradation
infrared
fluorescent probe
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN202011200558.6A
Other languages
Chinese (zh)
Other versions
CN112552507A (en
Inventor
刘世勇
周鑫
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
University of Science and Technology of China USTC
Original Assignee
University of Science and Technology of China USTC
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by University of Science and Technology of China USTC filed Critical University of Science and Technology of China USTC
Priority to CN202011200558.6A priority Critical patent/CN112552507B/en
Publication of CN112552507A publication Critical patent/CN112552507A/en
Application granted granted Critical
Publication of CN112552507B publication Critical patent/CN112552507B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
    • C08G65/00—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule
    • C08G65/02—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule from cyclic ethers by opening of the heterocyclic ring
    • C08G65/32—Polymers modified by chemical after-treatment
    • C08G65/329—Polymers modified by chemical after-treatment with organic compounds
    • C08G65/333—Polymers modified by chemical after-treatment with organic compounds containing nitrogen
    • C08G65/33396—Polymers modified by chemical after-treatment with organic compounds containing nitrogen having oxygen in addition to nitrogen
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
    • C08G65/00—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule
    • C08G65/02—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule from cyclic ethers by opening of the heterocyclic ring
    • C08G65/32—Polymers modified by chemical after-treatment
    • C08G65/329—Polymers modified by chemical after-treatment with organic compounds
    • C08G65/334—Polymers modified by chemical after-treatment with organic compounds containing sulfur
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
    • C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
    • C09K2211/14—Macromolecular compounds
    • C09K2211/1408—Carbocyclic compounds
    • C09K2211/1425—Non-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
    • C09K2211/14—Macromolecular compounds
    • C09K2211/1408—Carbocyclic compounds
    • C09K2211/1433—Carbocyclic compounds bridged by heteroatoms, e.g. N, P, Si or B
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
    • C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
    • C09K2211/14—Macromolecular compounds
    • C09K2211/1441—Heterocyclic
    • C09K2211/1466—Heterocyclic containing nitrogen as the only heteroatom
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G01N2021/6432—Quenching

Abstract

A trigger type self-degradation polymer-based near-infrared fluorescent probe, a preparation method and an application are provided, wherein the structural formula of the trigger type self-degradation polymer-based near-infrared fluorescent probe is as follows:
Figure DDA0002752701620000011
wherein the content of the first and second substances,
Figure DDA0002752701620000012
represents a trigger primitive;

Description

Trigger type self-degradation polymer-based near-infrared fluorescent probe, preparation method and application
Technical Field
The invention relates to the field of biomedical high polymer materials, in particular to a trigger type self-degradation polymer-based near-infrared fluorescent probe, a preparation method and application.
Background
Optical imaging has the advantages of being non-invasive and visualized and is therefore considered to be one of the most effective methods for studying the function and structure of cells or tissues. In recent years, near-infrared window imaging technology has been attracting much attention. Compared with the traditional visible light window imaging technology, the near infrared window imaging has the advantages of deep tissue penetration depth, good biocompatibility, tissue autofluorescence, tissue absorption, reduced tissue light scattering and the like, so the near infrared window imaging has higher space-time resolution and signal-to-background ratio (SBR). And because the near-infrared fluorophore can absorb near-infrared light, a photothermal effect can be generated for photothermal therapy (PTT); in addition, part of near-infrared fluorophores can transfer energy to surrounding oxygen under the excitation of light with specific wavelength to generate singlet oxygen with strong activity, the singlet oxygen can generate oxidation reaction with surrounding biomolecules, Glutathione (GSH) in cells is consumed to improve the intracellular oxidation pressure and accelerate apoptosis, and therefore photodynamic therapy (PDT) can be carried out, and the fluorophores can also be used for imaging-photothermal-photodynamic therapy synergistic action.
Accurate imaging of the tumor region is of great significance to diagnosis, monitoring and surgical navigation of the disease condition. However, most of the near-infrared window imaging reagents reported at present have non-specificity, that is, after intravenous injection into a body, the near-infrared window imaging reagents can not only be gathered in tumor tissues, but also be taken by other organs, so that strong interference signals exist, and accurate imaging of tumor regions is difficult to realize; most of the currently reported imaging agents are easily metabolized in vivo through the kidney or liver, and the long-term tracing effect is difficult to achieve, which is extremely disadvantageous to clinical diagnosis and surgical navigation. Therefore, the development of a near-infrared window imaging reagent which has high specificity, can trace for a long time and can realize synergistic treatment is of great significance.
Disclosure of Invention
In view of the above, the main objective of the present invention is to provide a triggered self-degradable polymer-based near-infrared fluorescent probe, a preparation method and applications thereof, so as to at least partially solve at least one of the above-mentioned technical problems.
In order to achieve the purpose, the technical scheme of the invention comprises the following steps:
as one aspect of the invention, a triggered self-degradation polymer-based near-infrared fluorescent probe is provided, and the structural formula is as follows:
Figure BDA0002752701600000021
wherein the content of the first and second substances,
Figure BDA0002752701600000022
represents a trigger primitive;
Figure BDA0002752701600000023
represents a near infrared fluorophore; the polymerization degree n is 5 to 10.
As another aspect of the present invention, there is also provided a method for preparing the triggered self-degradation polymer-based near-infrared fluorescent probe, comprising the following steps:
step 1: polymerizing a monomer containing phenoxycarbonyl and hydroxyl to form a polymer with chain ends comprising isocyanate groups and hydroxyl;
step 2: reacting a trigger unit with a terminal hydroxyl group with the polymer to generate a triggered self-degradable polymer;
and step 3: coupling the trigger type self-degradable polymer with polyethylene glycol acyl azide to obtain an amphiphilic block self-degradable polymer;
and 4, step 4: and reacting the amphiphilic block self-degradation polymer with a near-infrared fluorophore to obtain the triggered self-degradation polymer-based near-infrared fluorescent probe.
As a further aspect of the invention, the invention also provides an application of the triggered self-degradation polymer-based near-infrared fluorescent probe in the preparation of a tumor cell tracer.
Based on the technical scheme, compared with the prior art, the invention at least has the following beneficial effects:
the acid response triggered self-degradation polymer-based near-infrared probe provided by the invention has high-specificity imaging performance, and the near-infrared fluorophore with quinone methylene generated by degradation can be efficiently coupled with nucleophilic groups on the surface of proteins in tumor tissues, so that the problem that most of existing small molecular probes are rapidly metabolized in vivo is solved, and the long-term tracing effect is realized;
in addition, the photothermal and photodynamic properties of the near infrared fluorophore itself provide the possibility for the later-stage tumor synergistic treatment;
in conclusion, the acid response triggered self-degradation polymer-based near-infrared fluorescent probe provided by the invention can be used for long-term tracing and can be used for synergistic treatment.
Drawings
FIG. 1 is a nuclear magnetic hydrogen spectrum of a monomer containing a phenoxycarbonyl group and a hydroxyl group;
FIG. 2 is nuclear magnetic hydrogen spectra of amphiphilic block self-degradable polymer P2 and amphiphilic block self-degradable polymer P3 with side group post-modified near-infrared fluorophore in example 1 of the present invention;
FIG. 3 is the ultraviolet absorption spectrum of the amphiphilic block self-degradable polymer P3 of the near-infrared fluorescent small molecule IR820 and the side group post-modified near-infrared fluorophore in example 1 of the invention;
FIG. 4 is a transmission electron microscopy characterization of the assemblies obtained in example 1 of the present invention;
FIG. 5 shows the results of a light scattering particle size test of the assembly obtained in example 1 of the present invention;
FIG. 6 is a transmission electron microscopy characterization of the assembly obtained in comparative example 1;
FIG. 7 is a gel permeation chromatogram triggered by acidic conditions of the assembly in application example 1 of the present invention;
FIG. 8 is a graph showing an ultraviolet absorption spectrum triggered under an acidic condition of an assembly in practical example 1 of the present invention;
FIG. 9 shows the results of the scattering intensity triggered by acidic conditions of the assembly in example 1 of the present invention;
FIG. 10 is the result of the number of solution particles triggered by acidic conditions for the assembly of application example 1 of the present invention;
FIGS. 11a and 11b are the results of transmission electron microscope characterization of the assembly triggered by acidic conditions in application example 1 of the present invention, respectively, wherein FIG. 11a is the initial state; FIG. 11b is a state after a lapse of time;
FIG. 12 shows the results of fluorescence emission intensity changes triggered by acidic conditions for the assemblies of application example 1 of the present invention;
FIG. 13 shows the results of photothermal tests of the assembly of application example 2 of the present invention at different laser irradiation powers for the same polymer concentration;
FIG. 14 shows the results of photothermal tests of the assembly of application example 2 of the present invention at the same laser irradiation power and at different polymer concentrations;
FIG. 15 shows the results of the photodynamic performance test of the nanoparticles of application example 3 of the present invention in degraded and undegraded systems;
FIGS. 16a and 16b are statistics of cell viability after incubation of the assembly in application example 4 of the present invention with different types of normal cells, respectively, wherein FIG. 16a does not use 808nm laser irradiation; FIG. 16b uses 808nm laser irradiation;
FIGS. 17a, 17b and 17c are results of a photodynamic performance test of the assembly in application example 4 of the present invention at a cellular level, respectively; wherein, fig. 17a is a confocal image taken without the assembly but using 808nm laser irradiation, fig. 17b is a confocal image taken with the assembly but without using 808nm laser irradiation, and fig. 17c is a confocal image taken with the assembly and using 808nm laser irradiation;
FIG. 18 is a biological imaging result of the assembly in application example 5 of the present invention in a tumor-bearing mouse;
FIG. 19 is the result of bioimaging in tumor-bearing mice using the small molecule fluorophore IR820 in comparative example 1.
Detailed Description
The invention relates to a novel polymer-based near-infrared fluorescent probe capable of being traced for a long time, which can respond to a tumor part in a slightly acidic environment so as to realize specific fluorescence activation and long-term fluorescent tracing. The self-degradable polymers (SIPs) based on phenoxy carbonyl and hydroxyl connecting elements are designed and synthesized, and the terminal hydroxyl of the polymers and polyethylene glycol acyl azide are efficiently coupled to obtain the amphiphilic block copolymer. Near-infrared fluorophores are modified on the side groups thereof through efficient 'mercapto-maleimide' Michael addition reaction. After the amphiphilic block self-degradation polymer is chemically bonded with a near-infrared fluorophore, the amphiphilic block self-degradation polymer can self-assemble in an aqueous solution to form a nano particle, and the near-infrared fluorophore is gathered in a hydrophobic inner core to cause fluorescence quenching. Under physiological stimuli (e.g., pH less than 7), the polymer spontaneously depolymerizes in tandem and releases fluorescent molecules, thereby producing a fluorescent emission; the near-infrared fluorophore with quinone methylene produced by degradation can be efficiently coupled with nucleophilic groups on the surface of surrounding protein, thereby realizing long-term fluorescent tracing. In addition, the photothermal-photodynamic properties of the near-infrared fluorophore also have important application value for the synergistic treatment of tumors.
In order that the objects, technical solutions and advantages of the present invention will become more apparent, the present invention will be further described in detail with reference to the accompanying drawings in conjunction with the following specific embodiments.
As one aspect of the invention, a triggered self-degradation polymer-based near-infrared fluorescent probe is provided, and the structural formula is as follows:
Figure BDA0002752701600000051
wherein the content of the first and second substances,
Figure BDA0002752701600000052
represents a trigger primitive;
Figure BDA0002752701600000053
represents a near infrared fluorophore; the polymerization degree n is 5 to 10.
In the embodiment of the invention, the triggered self-degradation polymer-based near-infrared fluorescent probe is an amphiphilic block self-degradation polymer of a chemically bonded near-infrared fluorophore, which is composed of a hydrophilic polyethylene glycol chain segment and a hydrophobic acid-responsive self-degradation polymer chain segment with a pendant near-infrared fluorophore, and a Schiff base (T) is usednbs) As the end trigger primitive. Can realize fluorescence activation to the specific response of tumor site micro-acid environment, and the near-infrared fluorophore with quinone methylene group generated in the degradation process of the probe can efficiently capture the nucleophilic group on the surface of protein, thereby realizing long-term tracing effect. In addition, the photothermal-photodynamic properties of the near-infrared fluorophore also have important application value for the synergistic treatment of tumors.
In the embodiment of the invention, in the triggered self-degradation polymer-based near-infrared fluorescent probe, the polymerization degree n is too small, the number of modified fluorophores is small, and the high enough fluorescence intensity cannot be achieved; the polymerization degree n is too large, so that the nanoparticles assembled in the aqueous solution are too large and are not easy to enter tumor tissues and be taken up by tumor cells, and therefore, the polymerization degree n is suitable to be 5-10. More specifically, in embodiments of the present invention, the degree of polymerization, n, may be 5, 6, 7, 8, 9, or 10.
In an embodiment of the invention, the trigger primitive comprises an acid response trigger primitive;
in an embodiment of the present invention, the acidic response trigger primitive comprises:
Figure BDA0002752701600000061
in embodiments of the invention, the near-infrared fluorophore comprises indocyanine green or neoindocyanine green (IR 820).
In an embodiment of the invention, the triggered self-degradation polymer-based near-infrared fluorescent probe comprises a polymer obtained by polymerizing monomers containing phenoxycarbonyl and hydroxyl; the structural formula of the monomer containing the phenoxycarbonyl group and the hydroxyl group is as follows:
Figure BDA0002752701600000062
in the examples of the present invention, the monomer containing a phenoxycarbonyl group and a hydroxyl group, in carrying out polymerization reaction, forms a polymer whose chain end includes an isocyanate group and a hydroxyl group; the trigger unit with terminal hydroxyl is bonded with the isocyanate group of the polymer to generate the trigger type self-degradation polymer. The triggered self-degradable polymer can spontaneously carry out series depolymerization similar to domino after triggering and dissociating the protection element of a specific site, and simultaneously release a small molecule construction element. The trigger type self-degradation polymer-based near-infrared fluorescent probe containing the trigger type self-degradation polymer is degraded to generate a near-infrared fluorophore with quinone methylene, and can be efficiently coupled with nucleophilic groups on the surface of surrounding protein, so that long-term fluorescent tracing is realized.
In the examples of the present invention, the degradation process of the triggered self-degradable polymer-based near-infrared fluorescent probe exemplified by the near-infrared fluorophore IR820 and the specific structure of the near-infrared fluorophore with quinone methylene group are illustrated in the following reaction formula:
Figure BDA0002752701600000071
as another aspect of the present invention, there is also provided a method for preparing the above triggered self-degradable polymer-based near-infrared fluorescent probe, comprising the steps of:
step 1: polymerizing a monomer containing phenoxycarbonyl and hydroxyl to form a polymer with chain ends comprising isocyanate groups and hydroxyl;
step 2: reacting a trigger element with a terminal hydroxyl group with a polymer to generate a triggered self-degradable polymer;
and step 3: coupling the trigger type self-degradable polymer with polyethylene glycol acyl azide to obtain an amphiphilic block self-degradable polymer;
and 4, step 4: and reacting the amphiphilic block self-degradation polymer with a near-infrared fluorophore to obtain the triggered self-degradation polymer-based near-infrared fluorescent probe.
In an embodiment of the present invention, in step 1, the polymerization reaction comprises:
dissolving a monomer containing phenoxycarbonyl and hydroxyl in an organic solvent, adding a catalyst for catalysis in an oxygen-free atmosphere, and reacting for 2 to 5 hours at a temperature of between 70 and 90 ℃.
In the embodiment of the present invention, the oxygen-free atmosphere in step 1 may be, but is not limited to, a nitrogen atmosphere or an inert atmosphere.
In the embodiment of the present invention, dibutyltin dilaurate may be used as the catalyst in step 1.
In the embodiment of the present invention, the organic solvent in step 1 may be, but is not limited to, anhydrous dimethylsulfoxide.
In the embodiment of the present invention, the reaction temperature in step 1 may be, but not limited to, 70 ℃, 75 ℃, 80 ℃, 85 ℃, 90 ℃; the reaction time in step 1 may be, but is not limited to, 2 hours, 2.5 hours, 3 hours, 3.5 hours, 4 hours, 4.5 hours, 5 hours.
In an embodiment of the present invention, step 2 specifically includes:
adding a trigger element with a terminal hydroxyl group into the reaction liquid obtained in the step (1);
adding the trigger element and the monomer containing the phenoxycarbonyl group and the hydroxyl group according to an equimolar ratio;
reacting for 1-2 hours at 70-90 ℃;
after the reaction is finished, methanol is settled, N, N-dimethylformamide is dissolved, and repeated settlement and dissolution are carried out for multiple times; drying the obtained solid to obtain the solid trigger type self-degradation polymer.
In the embodiment of the present invention, the reaction temperature in step 2 may be the same as that in step 1; the reaction time in step 2 may be, but is not limited to, 1 hour, 1.5 hours, 2 hours.
In the present example, step 3, the reaction conditions were:
adding polyethylene glycol acyl azide and a triggered self-degradable polymer in a molar ratio of 2: 1-5: 1;
adding a catalyst for catalysis in an organic solvent under an oxygen-free atmosphere, and reacting at 70-90 ℃ for 10-15 hours;
after the reaction is finished, methanol is settled, N, N-dimethylformamide is dissolved, and repeated settlement and dissolution are carried out for multiple times; and drying the obtained solid to obtain the solid amphiphilic block self-degradation polymer.
In embodiments of the present invention, the molar ratio of the polyethylene glycol acyl azide to the triggered self-degrading polymer in step 3 may be, but is not limited to, 2: 1, 3: 1, 4: 1, 5: 1.
In the embodiment of the present invention, the organic solvent in step 3 may be, but is not limited to, dimethyl sulfoxide.
In the embodiment of the present invention, the oxygen-free atmosphere in step 3 may be a nitrogen atmosphere or an inert atmosphere.
In the embodiment of the present invention, the catalyst in the step 3 may be, but is not limited to, dibutyltin dilaurate.
In the embodiment of the present invention, the reaction temperature in step 3 may be, but not limited to, 70 ℃, 75 ℃, 80 ℃, 85 ℃, 90 ℃; the reaction time may be, but is not limited to, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours.
In an embodiment of the present invention, step 4 specifically includes:
carrying out Michael addition reaction on the amphiphilic block self-degradation polymer and a near-infrared fluorophore;
carrying out self-assembly reaction to obtain a triggered self-degradation polymer-based near-infrared fluorescent probe of the nano particles;
wherein the Michael addition reaction comprises:
adding a near-infrared fluorophore and an amphiphilic block self-degradation polymer in a proportion of 1: 6-1: 10;
reacting in an organic solvent in a nitrogen atmosphere at 25-35 ℃ for 12-24 hours;
after the reaction is finished, methanol is settled, N, N-dimethylformamide is dissolved, and repeated settlement and dissolution are carried out for multiple times; drying the obtained solid to obtain a solid trigger type self-degradation polymer-based near-infrared fluorescent polymer;
in the embodiment of the present invention, the organic solvent in the step 4 may be, but not limited to, N-dimethylformamide.
In the embodiment of the present invention, the reaction temperature in step 4 may be, but is not limited to, 25 ℃, 30 ℃, 35 ℃; the reaction time may be, but is not limited to, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours.
In the embodiment of the present invention, the molar ratio of the near-infrared fluorophore to the amphiphilic block self-degradable polymer in the step 4 can be, but is not limited to, 1: 6, 1: 7, 1: 8, 1: 9, and 1: 10.
Wherein the self-assembly reaction comprises:
dissolving a solid trigger type self-degradation polymer-based near-infrared fluorescent polymer in an organic solvent, and adding water under the stirring condition;
and (4) dialyzing to remove the organic solvent to obtain the nano particle triggered self-degradation polymer-based near-infrared fluorescent probe.
In the embodiment of the present invention, the organic solvent for the self-assembly reaction may be, but is not limited to, dimethyl sulfoxide.
In the embodiment of the invention, the concentration of the triggered self-degradation polymer-based near-infrared fluorescent probe of the nano particles is 0.1-1 mg/mL.
More specifically, the concentration of the triggered self-degradation polymer-based near-infrared fluorescent probe of the nanoparticles is too high, and the particle size distribution of the obtained nanoparticles is not uniform; the concentration is too low, and the application effect is not obvious. Therefore, it is preferably 0.1 to 1 mg/mL; preferably, the concentration is 0.2 mg/mL.
As a further aspect of the invention, the invention also provides an application of the triggered self-degradation polymer-based near-infrared fluorescent probe in the preparation of a tumor cell tracer.
The technical solution of the present invention is further described below with reference to specific examples, but it should be noted that the following examples are only for illustrating the technical solution of the present invention, but the present invention is not limited thereto.
Example 1
In the first step, the introduction of the trigger can be accomplished by adding an alcoholic hydroxyl group-containing trigger during the polymerization. To assist understanding more clearly, the following example is given for the protection of the triggered self-degradable polymer (P1) with 4-nitrobenzylideneaminobenzol. The reaction formula is shown as follows:
Figure BDA0002752701600000101
the polymerization degree n of the obtained polymer is between 5 and 10.
The preparation method comprises the following steps: a monomer containing a phenoxycarbonyl group and a hydroxyl group (0.5g, 0.94mmol), dibutyltin Dilaurate (DBTL) (5.9mg, 0.0094mmol) and anhydrous dimethyl sulfoxide (DMSO) (1mL) were placed in a 10mL reaction flask, evacuated and charged with nitrogen gas to place the system in a nitrogen atmosphere, and then reacted at 85 ℃ for 3 hours. 4-Nitrobenzylideneaminobenzyl alcohol (0.24g, 0.94mmol) was added and the reaction was continued at 85 ℃ for 1 h. After the reaction was completed, the reaction mixture was precipitated in methanol, dissolved in N, N-dimethylformamide, reprecipitated, redissolved, and repeated 3 times, and the resulting solid was dried in a vacuum oven overnight to finally obtain a product having a polymerization degree N of 6 (0.38g, yield 84%) as a pale yellow solid.
Secondly, synthesizing an amphiphilic block self-degradation polymer, wherein the reaction general formula is shown as the following formula:
Figure BDA0002752701600000102
the preparation method comprises the following steps: mixing PEG-CON3(0.35g, 0.16mmol), P1(0.2g, 0.04mmol), DBTL (0.5mg, 0.0004mmol) and 1mL of anhydrous DMSO were placed in a 10mL reaction flask, evacuated and purged with nitrogen to place the system under a nitrogen atmosphere, and then reacted at 85 ℃ for 15 h. After the reaction was completed, the reaction mixture was settled in methanol, dissolved in N, N-dimethylformamide, re-settled, re-dissolved, and repeated 3 times, and the obtained solid was dried overnight in a vacuum oven to finally obtain a product (0.25g, yield 82%) as a pale yellow solid.
The third step: modifying the near-infrared fluorescent molecule through a mercapto-double bond Michael addition reaction, wherein the general reaction formula is shown as the following formula:
Figure BDA0002752701600000111
the preparation method comprises the following steps: polymer P2(30mg, 1eq), IR820-SH (20mg, 20eq) and N, N-Dimethylformamide (DMF) were placed in a 10mL reaction flask, evacuated and charged with nitrogen gas to place the reaction system under a nitrogen atmosphere, and then reacted at room temperature for 12 hours. After the reaction is finished, settling the reaction mixture in methanol, dissolving the N, N-dimethylformamide, and repeatedly settling and dissolving for 3 times; the resulting solid was dried in a vacuum oven overnight to give the product (32mg, 89% yield) as a dark green solid.
The fourth step: preparation of nanoparticles SIP-NPs
Polymer P3(2mg) was dissolved in 1mL of DMSO, and then 8mL of water was added slowly under magnetic stirring (500rpm) at a rate of 1 mL/h. After the water is added, a dialysis bag with the cut-off molecular weight of 1 and 4000Da is used for dialysis in deionized water to remove the organic solvent in the system, the water is changed once every 6 hours for 3 times in total, after the dialysis is finished, the system solution (water) is supplemented to 10mL, and the concentration of the finally obtained assembly is 0.2 mg/mL.
The triggered self-degradation polymer-based near-infrared fluorescent probe obtained in this example 1 and the raw materials or intermediates in the respective steps were characterized, and the following results were obtained.
(1) FIG. 1 shows nuclear magnetic hydrogen spectra of the starting monomers participating in the reaction in the first step. As shown in fig. 1, the phenoxy carbonyl group, the hydroxyl group, and the hydrogen corresponding to the functional group of the carbon-carbon double bond to be subjected to the michael addition reaction contained in the monomer all have one-to-one correspondence in the nuclear magnetic hydrogen spectrum, so that the correctness of the monomer structure can be proved.
(2) FIG. 2 is nuclear magnetic hydrogen spectra of amphiphilic block self-degradable polymer P2 and amphiphilic block self-degradable polymer P3 with side group post-modified near infrared fluorophore. As shown in FIG. 2, through comparison of nuclear magnetic hydrogen spectra of P2 and P3, the newly added hydrogen of the amphiphilic block self-degradation polymer P3 with the side group post-modified is contributed by IR820 near-infrared fluorescent molecules. Thus, the amphiphilic block self-degradation polymer P2 is successfully modified to be provided with an IR820 near infrared fluorescent molecule.
(3) FIG. 3 is an ultraviolet absorption spectrum of nanoparticles assembled by near-infrared fluorescent micromolecules IR820 and amphiphilic block self-degradation polymers P3 with side groups modified later dissolved in DMSO solvent and P3 in aqueous solution. As shown in fig. 3, the absorption peak after modification of IR820 onto the polymer substantially coincided with the small molecule IR820, thereby demonstrating the structural integrity of IR 820. In addition, the absorption spectrum of the nanoparticle shows a new absorption peak, which indicates that in the nanoparticle state, the IR820 near infrared fluorophore is wrapped in the hydrophobic core and aggregation occurs.
(4) Fig. 4 and 5 are a Transmission Electron Microscope (TEM) characterization image and a light scattering (ALV) characterization image, respectively, of the assembly prepared in example 1.
FIG. 4 is the TEM test result of a sample having a polymerization degree of 6 in example 1, and it can be seen that the obtained nanoparticles have a very uniform spherical structure with a diameter of about 70 nm; FIG. 5 shows the ALV test results of the sample with polymerization degree of 6 in example 1, and the measured particle size of the nanoparticles is 78nm (since ALV is a solution system, and TEM is dry, TEM usually has smaller particle size than ALV), which is consistent with TEM results. Thus, the sample having a polymerization degree of 6 in example 1 yielded uniform spherical nanoparticles.
Example 2
Example 2 the same preparation as in example 1 was used except that:
the reaction conditions of the first step include: dissolving a monomer containing phenoxycarbonyl and hydroxyl in an organic solvent, adding dibutyltin dilaurate for catalysis in an inert gas atmosphere, and reacting for 2.5 hours at 70 ℃. The trigger element is added according to the molar ratio of the trigger element to the monomers of the phenoxycarbonyl group and the hydroxyl group; the reaction was carried out at 90 ℃ for 1 hour.
The polymerization degree of the finally obtained trigger type self-degradation polymer-based near-infrared fluorescent probe is 5.
The triggered self-degradation polymer-based near-infrared fluorescent probe finally obtained in the example 2 is spherical nano-particles with uniform particle size.
Example 3
Example 3 the same preparation method as example 1 was used, except that:
the reaction conditions of the first step include: dissolving a monomer containing phenoxycarbonyl in an organic solvent, adding dibutyltin dilaurate for catalysis in an inert gas atmosphere, and reacting for 4 hours at 90 ℃. The trigger element is added according to the molar ratio of the trigger element to the monomers of the phenoxycarbonyl group and the hydroxyl group; the reaction was carried out at 90 ℃ for 2 hours.
The polymerization degree of the finally obtained trigger type self-degradation polymer-based near-infrared fluorescent probe is 10.
The triggered self-degradation polymer-based near-infrared fluorescent probe finally obtained in the example 3 is spherical nano-particles with uniform particle size.
Comparative example 1
This comparative example 1 was prepared as in example 1, except that:
the reaction conditions of the first step include: dissolving a monomer containing phenoxycarbonyl in an organic solvent, adding dibutyltin dilaurate for catalysis in an inert gas atmosphere, and reacting for 4 hours at 100 ℃. The trigger element is added according to the molar ratio of the trigger element to the monomers of the phenoxycarbonyl group and the hydroxyl group; the reaction was carried out at 100 ℃ for 2 hours.
The polymerization degree of the finally obtained trigger type self-degradation polymer-based near-infrared fluorescent probe is 13.
Fig. 6 is TEM test results of the sample having the polymerization degree of 13 in comparative example 1, and as shown in fig. 6, it can be seen that the nanoparticles obtained in comparative example 1 have no stable morphology and very non-uniform particle size.
The following application examples 1 to 5 are all described by taking the triggered self-degradable polymer-based near-infrared probe nanoparticles obtained in example 1 as an example.
Application example 1 acidic pH triggers nanoparticle degradation and enables fluorescence activation
The tumor tissue is in a slightly acidic environment (pH is 6.0-6.5), and the pH inside lysosomes of tumor cells can reach 5.0. Based on this, 1mL of the assembly of example 1 with a concentration of 0.2mg/mL was taken into a 10mL sample bottle, 1mL of a phosphate buffer solution (pH 6.0, 200mM) was added, 0.2mL of an aqueous solution of Glutathione (GSH) with a concentration of 100mM was added, the mixed system was incubated at 37 ℃, samples were taken at fixed time points and monitored, depolymerization of the molecular chain level after triggering was measured by ultraviolet absorption spectroscopy and gel permeation chromatography, changes in the physical properties of the polymer nanoparticles after triggering depolymerization were observed by static light scattering (ALV), nanoparticle tracking analyzer (Nanosight), transmission electron microscope, and the degradation of the polymer and the fluorescence emission were tracked by fluorescence spectroscopy.
As shown in fig. 7 to fig. 12, the experimental results show that the triggered self-degradable polymer-based near-infrared fluorescent probe can be triggered and serially depolymerized under an acidic pH condition, and the molecular weight of the polymer is obviously gradually decreased according to gel permeation chromatography (fig. 7); the uv absorption spectrum also seen a gradual red shift in the absorption maxima of the polymer with increasing degradation time (fig. 8), thereby confirming that depolymerization of the polymer occurred at the molecular chain level. As the polymer is degraded into small molecular fragments, the Bovine Serum Albumin (BSA) in the system captures quinone methylene generated by degradation to form water-soluble small molecular fragments, so that the nanoparticles gradually collapse and decompose, the ALV tracking finds that the scattering light intensity of the nanoparticle solution is gradually reduced, and the nano sight test result further proves the depolymerization of the nanoparticles (FIG. 9 and FIG. 10). The number of nanoparticles in the degradation system was gradually reduced by transmission electron microscopy until finally almost no large aggregates could be seen (fig. 11a and 11 b). Fluorescence spectroscopy tests found that the nanoparticle state had no fluorescence emission, while the fluorescence emission intensity gradually increased during the slightly acidic environment triggering polymer degradation (fig. 12).
In conclusion, the results fully illustrate the feasibility of the triggered self-degradation polymer-based near-infrared fluorescent probe, which can specifically respond to a slightly acidic environment and realize near-infrared fluorescence emission activation.
Application example 2 photothermal Properties of Polymer fluorescent Probe nanoparticles under 808nm laser irradiation
Preparing a nanoparticle aqueous solution of the assembly obtained in example 1 with an IR820 dye concentration of 25 μ M, irradiating a sample for 5min under different powers by using a 808nm laser, and monitoring and recording the temperature of the system in real time by using a digital temperature monitor. The experimental result shows that the laser power is 3W/cm2Although the temperature of the system can be raised to the maximum, the power is too strong, and the bleaching of the dye is serious. 1W/cm2The irradiation power of the system can not only increase the temperature of the system to be more than 60 ℃ but also can not cause the bleaching of the dye, so that 1W/cm is selected subsequently2The experiment was carried out (fig. 13). Determining the irradiation power to be 1W/cm2Thereafter, the nanoparticle solutions of the assemblies obtained in example 1, in which the IR820 dye concentrations were 5. mu.M, 10. mu.M, and 25. mu.M, respectively, were tested for photothermal effect, and it was found that the photothermal effect gradually increased with the increase in the concentration (FIG. 14). Since the temperature is above 60 ℃, the tumor cells can be killed well, so that the concentration of IR820 is 25 μ M, which is suitable for killing tumor cells.
Application example 3 photodynamic Properties of Polymer fluorescent Probe nanoparticles before and after degradation
The photodynamic performance of the dye molecules in a dispersed state is better than that in an aggregation state, so the difference of the photodynamic performance before and after degradation can reflect the degradation degree of the system. Based on this, Dichlorodihydrofluorescein (DCFH) was used as the active oxygen detection reagent, and the undegraded system was maintained at pH 7.4, IR820 concentration 15. mu.M, and DCFH concentration 10. mu.M. The pH of the degradation system is adjusted to 6.0, and then30 μ M BSA was used to capture the quinone methylene, IR820 concentration 15 μ M DCFH concentration 10 μ M, the system was incubated at 25 ℃ for 24h and then tested, using 808nm laser irradiation at 1W/cm irradiation power2. The test results found that the fluorescence intensity of DCFH in the degraded system was stronger than that in the undegraded system (FIG. 15). Meanwhile, the excellent photodynamic performance of the probe is verified to be applicable to elimination of cancer cells, and the probe is very expected to be combined with photothermal effect for treatment of tumors.
Application example 4 cytotoxicity and photodynamic Properties at cellular level of polymeric fluorescent probes
The tumor cells have slightly acidic environment, so the probe obtained in the example 1 can be triggered to depolymerize after being co-incubated with the tumor cells, and the effect of killing the cancer cells can be well realized by adding singlet oxygen generated by laser irradiation at 808nm and photothermal effect. Based on this, 4T1, 3T3, LO2, HepG2 and HeLa cells were seeded in 96-well plates at a density of 5000 cells per well, 100mL of complete cell culture medium (DMEM) was added, the culture medium was incubated at 37 ℃ for 24h, replaced with fresh DMEM medium, different concentrations of the assembly of example 1 were added (40. mu.M, 20. mu.M, 10. mu.M, 5. mu.M, 2.5. mu.M, 1.25. mu.M, 0.625. mu.M, 0.3. mu.M, respectively, calculated as the concentration of IR820), incubation was continued for 24h at 37 ℃ followed by addition of thiazolyl blue tetrazolium bromide (MTT) reagent (20. mu.L, 5mg/mL of Phosphate Buffered Saline (PBS) solution) per well for 4h, the medium was removed, 100. mu.L of DMSO was added, shaking was continued for 15min, absorbance at 490nm was recorded using a microplate reader, the results are shown in FIG. 16a and FIG. 16b, where the concentration of the five different cell assemblies were shown in FIG. 16a, cell viability results without 808nm laser irradiation; FIG. 16b is the cell viability results of five different cells incubated with different concentrations of assemblies and irradiated with 808nm laser. Experimental results show that the probe has almost no toxicity to normal cells before and after 808nm laser irradiation, but has outstanding toxicity to cancer cells, and the cytotoxicity is greatly enhanced after illumination.
To further verify the contribution of photodynamic properties in cytotoxicity, HeLa cells were selected as a model, per cellHole 2X105The seeds were incubated at 37 ℃ for 24h in glass four-well plates, the medium was removed, 200. mu.L of the medium containing the assemblies of example 1 (IR820 concentration 20. mu.M) was added to two wells, 200. mu.L of complete DMEM medium was added to the remaining two wells, incubation was performed at 37 ℃ for 4h, the medium containing the assemblies was removed, the cells were washed 3 times with PBS, 200. mu.L of the medium containing 2 ', 7' -dichlorofluorescein diacetate (DCFH-DA) (5. mu.M) was added, and incubation was continued at 37 ℃ for 30 min. The medium was removed, washed 2 times with PBS, 100. mu.L of PBS was added to each well, and irradiated with 808nm laser for 5min (1W/cm)2) Confocal imaging was immediately performed. Fig. 17a is a confocal image taken with no assembly but with 808nm laser irradiation, fig. 17b is a confocal image taken with assembly but without 808nm laser irradiation, and fig. 17c is a confocal image taken with assembly and with 808nm laser irradiation. The results show that the cells (SIP-NPs + light) in the light group show strong DCFH-DA fluorescence (FIG. 17c), thereby proving that the probe also has excellent photodynamic performance at the cellular level, and the visible photodynamic effect plays an important role in the process of killing tumor cells.
Application example 5 tumor site-specific imaging and Long-term tracking of tumor-bearing mice
In vivo experiments are the decisive factor for verifying the material performance, and based on the fact, 200 mu L of the assembly of example 1 with the concentration of 0.5mg/mL is taken, and Balb/c (6-7 weeks) nude mice with tumors are injected into tail veins, and the fluorescence distribution of the whole bodies of the mice is shot at fixed time points by using a living animal imaging system. As shown in fig. 18, the results show that after 48h of tail vein injection, fluorescence can be seen almost only at the tumor site, and the tumor boundary can be clearly seen, so that the method is very promising for accurate surgical navigation tumor resection. Furthermore, most notably, the fluorescence intensity at the tumor site remained at 40% by day 8. This result confirms that the quinone methylene group generated by the degradation of the polymer probe can effectively capture the nucleophilic group on the surface of the surrounding protein in the tumor tissue.
Application comparative example 1
A small molecular fluorophore IR 820200 mu L with the concentration of 0.5mg/ml is taken, a Balb/c (6-7 weeks) nude mouse with a tumor is injected into a tail vein, and the fluorescence distribution of the whole body of the mouse is shot at fixed time points by using a living body imaging system of the mouse. As shown in fig. 19, the results showed that the small molecule fluorophore IR820 was completely metabolized well 4h after tail vein injection and little fluorescence emission was seen at the tumor site.
The above-mentioned embodiments are intended to illustrate the objects, technical solutions and advantages of the present invention in further detail, and it should be understood that the above-mentioned embodiments are only exemplary embodiments of the present invention and are not intended to limit the present invention, and any modifications, equivalents, improvements and the like made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (9)

1. A triggered self-degradation polymer-based near-infrared fluorescent probe is characterized in that the structural formula is as follows:
Figure FDA0003176605370000011
wherein the content of the first and second substances,
Figure FDA0003176605370000012
represents a trigger primitive; wherein the trigger primitive comprises an acid response trigger primitive; wherein the acid response trigger primitive comprises:
Figure FDA0003176605370000013
wherein the content of the first and second substances,
Figure FDA0003176605370000014
represents a near infrared fluorophore; wherein the near-infrared fluorophore comprises indocyanine green or neoindocyanine green; the polymerization degree n is 5 to 10.
2. The triggered self-degrading polymer-based near-infrared fluorescent probe according to claim 1, wherein the triggered self-degrading polymer-based near-infrared fluorescent probe comprises a polymer obtained by polymerizing a monomer containing a phenoxycarbonyl group and a hydroxyl group; the structural formula of the monomer containing the phenoxycarbonyl and the hydroxyl is as follows:
Figure FDA0003176605370000015
3. a method for preparing the triggered self-degradable polymer-based near-infrared fluorescent probe according to any one of claims 1 to 2, comprising the following steps:
step 1: polymerizing a monomer containing phenoxycarbonyl and hydroxyl to form a polymer with chain ends comprising isocyanate groups and hydroxyl;
step 2: reacting a trigger unit with a terminal hydroxyl group with the polymer to generate a triggered self-degradable polymer;
and step 3: coupling the trigger type self-degradable polymer with polyethylene glycol acyl azide to obtain an amphiphilic block self-degradable polymer;
and 4, step 4: and reacting the amphiphilic block self-degradation polymer with a near-infrared fluorophore to obtain the triggered self-degradation polymer-based near-infrared fluorescent probe.
4. The method of claim 3, wherein in step 1, the polymerization reaction comprises:
dissolving a monomer containing phenoxycarbonyl and hydroxyl in an organic solvent, adding a catalyst for catalysis in an oxygen-free atmosphere, and reacting for 2 to 5 hours at a temperature of between 70 and 90 ℃.
5. The method according to claim 4, wherein the step 2 comprises:
adding a trigger element with a terminal hydroxyl group into the reaction liquid obtained in the step 1;
the trigger element and the monomer containing the phenoxyl carbonyl group and the hydroxyl group are added according to an equal molar ratio;
reacting for 1-2 hours at 70-90 ℃;
after the reaction is finished, methanol is settled, N, N-dimethylformamide is dissolved, and repeated settlement and dissolution are carried out for multiple times; drying the obtained solid to obtain the solid trigger type self-degradation polymer.
6. The method according to claim 3, wherein in the step 3, the reaction conditions are as follows:
polyethylene glycol acyl azide and a triggered self-degrading polymer are mixed in a molar ratio of 2: 1-5: 1, adding a proportional relation;
adding a catalyst for catalysis in an organic solvent under an oxygen-free atmosphere, and reacting at 70-90 ℃ for 10-15 hours;
after the reaction is finished, methanol is settled, N, N-dimethylformamide is dissolved, and repeated settlement and dissolution are carried out for multiple times; and drying the obtained solid to obtain the solid amphiphilic block self-degradation polymer.
7. The method according to claim 3, wherein the step 4 comprises:
carrying out Michael addition reaction on the amphiphilic block self-degradation polymer and a near-infrared fluorophore;
carrying out self-assembly reaction to obtain a triggered self-degradation polymer-based near-infrared fluorescent probe of the nano particles;
wherein the Michael addition reaction comprises:
mixing a near-infrared fluorophore and an amphiphilic block self-degradation polymer in a molar ratio of 1: 6-1: 10, adding a proportional relation;
reacting in an organic solvent in a nitrogen atmosphere at 25-35 ℃ for 12-24 hours;
after the reaction is finished, methanol is settled, N, N-dimethylformamide is dissolved, and repeated settlement and dissolution are carried out for multiple times; drying the obtained solid to obtain a solid trigger type self-degradation polymer-based near-infrared fluorescent polymer;
wherein the self-assembly reaction comprises:
dissolving the solid trigger type self-degradation polymer-based near-infrared fluorescent polymer in an organic solvent, and adding water under the stirring condition;
and (4) dialyzing to remove the organic solvent to obtain the nano particle triggered self-degradation polymer-based near-infrared fluorescent probe.
8. The preparation method of claim 7, wherein the concentration of the triggered self-degradation polymer-based near-infrared fluorescent probe of the nanoparticle is 0.1-1 mg/mL.
9. Use of the triggered self-degrading polymer-based near-infrared fluorescent probe according to any one of claims 1 to 2 in the preparation of a tumor cell tracer.
CN202011200558.6A 2020-10-30 2020-10-30 Trigger type self-degradation polymer-based near-infrared fluorescent probe, preparation method and application Active CN112552507B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202011200558.6A CN112552507B (en) 2020-10-30 2020-10-30 Trigger type self-degradation polymer-based near-infrared fluorescent probe, preparation method and application

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202011200558.6A CN112552507B (en) 2020-10-30 2020-10-30 Trigger type self-degradation polymer-based near-infrared fluorescent probe, preparation method and application

Publications (2)

Publication Number Publication Date
CN112552507A CN112552507A (en) 2021-03-26
CN112552507B true CN112552507B (en) 2021-10-01

Family

ID=75042525

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202011200558.6A Active CN112552507B (en) 2020-10-30 2020-10-30 Trigger type self-degradation polymer-based near-infrared fluorescent probe, preparation method and application

Country Status (1)

Country Link
CN (1) CN112552507B (en)

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ES2402614B1 (en) * 2011-10-24 2014-03-18 Centro De Investigación Principe Felipe CONTROLLED SYNTHESIS OF POLYGLUTAMATES WITH LOW POLIDISPERSITY AND VERSATILE ARCHITECTURES
CN103588940B (en) * 2013-11-07 2016-01-20 中国科学技术大学 Amphipathilic block polymer, polymer vesicle and its preparation method and application

Also Published As

Publication number Publication date
CN112552507A (en) 2021-03-26

Similar Documents

Publication Publication Date Title
CN108864106B (en) The preparation and application of two area's small organic molecule fluorescence probe of near-infrared
CN113943305B (en) Phthalocyanine fluorescent molecular compound, preparation method and phthalocyanine fluorescent nano material
CN109529034B (en) Near-infrared two-region conjugated nano particle and preparation method and application thereof
CN110041475B (en) Amphiphilic block copolymer, shell-crosslinked micelle thereof, preparation method and application
CN110997012A (en) Polymer nanoparticles for afterglow molecular imaging
CN113773667B (en) Organic small molecule near infrared two-region fluorescent dye and preparation method and application thereof
CN113087877B (en) Near-infrared two-region fluorescence emission water-soluble conjugated polymer nano phototherapy reagent and preparation method and application thereof
CN107469079B (en) Preparation method of photodynamic therapeutic agent under guidance of T1-MRI imaging
CN110302177B (en) Mitochondrion targeting photo-diagnosis nano-particle and application thereof
WO2019136866A1 (en) Preparation of novel photosensitizer composite nano multifunctional material and use thereof
CN109453394B (en) Probe based on mesoporous carbon nanosphere doped gold nanoparticle material and preparation method thereof
CN112494663B (en) Multi-modal nano diagnosis and treatment reagent and preparation method and application thereof
KR101183732B1 (en) Conjugate of acetylated polysccharide and photosensitizer for photodynamic diagnosis or therapy and process of preparation thereof
Liu et al. Thiophene donor for nir-ii fluorescence imaging-guided photothermal/photodynamic/chemo combination therapy
Zhang et al. Hierarchical nano-to-molecular disassembly of boron dipyrromethene nanoparticles for enhanced tumor penetration and activatable photodynamic therapy
CN113827722B (en) Controllable preparation of ONOO - Nanoparticle of (a) and preparation method and application thereof
Zhang et al. A single-wavelength NIR-triggered polymer for in situ generation of peroxynitrite (ONOO−) to enhance phototherapeutic efficacy
CN109320636B (en) Triple stimulus-responsive core-crosslinked polymer micelle and preparation method and application thereof
CN113082206B (en) Macromolecule nitric oxide donor modified up-conversion nanoparticle, preparation method and application
Huang et al. A hierarchical supramolecular nanozyme platform for programming tumor-specific PDT and catalytic therapy
CN112552507B (en) Trigger type self-degradation polymer-based near-infrared fluorescent probe, preparation method and application
CN111333819B (en) Compound and application thereof
Yan et al. Single-laser excitation synergistic photo-and chemodynamic therapy system based on persistent luminescence nanoparticles
CN113773488A (en) Preparation method of water-soluble polyporphyrin carrier-free nano-drug
CN114848852B (en) Self-assembled graphene quantum dot nanoparticle, and preparation method and application thereof

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant