CN112679478B - Near-infrared two-region fluorescent molecular probe of targeted tyrosine kinase and preparation method and application thereof - Google Patents
Near-infrared two-region fluorescent molecular probe of targeted tyrosine kinase and preparation method and application thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of biomedicine, and particularly provides a near-infrared two-region fluorescent molecular probe of targeted tyrosine kinase, a preparation method and application thereof, and the near-infrared two-region fluorescent molecular probe is applied to tumor targeted fluorescence imaging and is formed by connecting a tumor tyrosine kinase identification group (TKP), a near-infrared two-region fluorescent group (SQ1050) and a tumor targeted polypeptide group (PEP) through chemical bonds. The reagent is based on the high expression of tyrosine kinase of tumors, can be applied to tumors with high expression of tyrosine kinase, and can be applied to tumor targeted fluorescence imaging, dynamic tracing of tumor tyrosine kinase and the like by combining a near-infrared two-region fluorescence imager. The reagent realizes high sensitivity and high specificity in application, and provides an effective tool for tumor research in medicine and clinical monitoring and treatment of tumors, so the reagent has good application prospects in the aspects of tumor targeted fluorescence imaging, tumor tyrosine kinase dynamic tracing and the like.
Description
Technical Field
The invention belongs to the technical field of biomedicine, and relates to a detection reagent, a preparation method and application thereof, in particular to a near-infrared two-region fluorescent probe of targeted tyrosine kinase, a preparation method thereof and application for tumor targeted fluorescence imaging and monitoring of tumor tyrosine kinase content.
Background
Tyrosine kinases are high affinity cell surface receptors for many growth factors, cytokines and hormones. Of the 90 unique tyrosine kinase genes identified in the human genome, 58 encode receptor tyrosine kinase proteins. Tyrosine kinases have an extracellular receptor domain, and an extracellular signaling substance binds to the receptor moiety and activates its intracellular kinase activity domain, thereby phosphorylating tyrosine residues of the substrate. Receptor tyrosine kinases have not only been shown to be key regulators of normal cellular processes, but also play a key role in the development and progression of many types of cancer. Mutations in receptor tyrosine kinases activate a series of signaling cascades that have many effects on protein expression. After the ligand is combined, the tyrosine kinase can generate spontaneous polymerization, activate the kinase activity to self-phosphorylate and conduct to a plurality of signal paths in cells, such as MAPK, PI3K-Akt signal paths and the like. Since the discovery of tyrosine kinases, a large body of clinical data has demonstrated a significant impact on a variety of diseases, particularly cancer. Tyrosine kinases undergo extensive amplification and multiple site mutations in cancer tissues. In recent years, with the rapid development of precision medicine, molecular targeted drugs against tyrosine kinases have begun to enter the market. There is still a great need for further research in various aspects of tyrosine kinases.
Small molecule fluorescent probes are widely used in chemical analysis, biological analysis and real-time, non-invasive monitoring of biodynamic processes. Therefore, the small-molecule fluorescent probe is widely used for drug screening and treatment effect evaluation, and early monitoring of tumor treatment effect is realized. The newly developed near-infrared two-zone fluorescent molecular imaging technology breaks through the limitation of the penetration depth of optical tissues and can obtain a fluorescent image with higher optical resolution than the traditional optical resolution at the living body level.
Disclosure of Invention
Aiming at the problems, the invention aims to prepare a near-infrared two-region fluorescent molecular probe targeting tyrosine kinase, a preparation method and application thereof. After the molecular probe is combined with tyrosine kinase, the fluorescence intensity of the near-infrared two-region is obviously enhanced, the fluorescence can be detected by a near-infrared two-region fluorescence detector, and the content and the distribution of the tyrosine kinase in various biological systems can be evaluated by utilizing the reaction of the molecular probe.
The invention also provides a reagent for tumor targeted near-infrared two-zone fluorescence imaging and tumor metastasis monitoring, which can be used as a reagent for surgical navigation.
The third purpose of the invention is to provide the application of the near-infrared two-region fluorescent molecular probe targeting tyrosine kinase in preparing a reagent for tumor targeting near-infrared two-region fluorescence imaging and tumor metastasis monitoring.
The fourth purpose of the invention is to provide the application of the near-infrared two-region fluorescent molecular probe targeting the tyrosine kinase in preparing a reagent for monitoring the content and distribution of the tumor tyrosine kinase.
The technical scheme of the invention is as follows:
the near-infrared two-region fluorescent molecular probe of the targeted tyrosine kinase is formed by connecting a tyrosine kinase identification group (TKP) and a near-infrared two-region fluorescent group (SQ1050) through chemical bonds; the structure is as follows: a structure I type TKP-SQ1050 or a structure II type TKP-SQ 1050-PEP.
The tyrosine kinase recognition group (TKP) has the structure:
the structure of the near-infrared two-region fluorescent group (SQ1050) is as follows:
preferably, the structure of the near-infrared two-region fluorescent probe targeting the tyrosine kinase is shown as the following formula TFPSQ-1:
the near-infrared two-region fluorescent probe targeting tyrosine kinase preferably has the structure shown in the following formula TFPSQ-2, wherein the structure of the TKP-SQ1050-PEP with the structure II is shown in the specification:
the near-infrared two-region fluorescent probe of the targeted tyrosine kinase has low fluorescence quantum yield, has high fluorescence quantum yield after the molecular probe is combined with the tyrosine kinase, and can realize the rapid and sensitive detection of the content and the distribution of the tyrosine kinase by adopting a fluorescence detector.
The probe does not have near-infrared two-region fluorescence, the probe formed by combining a tyrosine kinase recognition group in the probe and tyrosine kinase has strong near-infrared two-region fluorescence, and a fluorescence detector can be used for realizing rapid and sensitive detection of substrate fluorescence change.
The probe is used as a ligand of the tyrosine kinase, and the content of the tyrosine kinase in different biological systems is quantitatively determined by quantitatively detecting the fluorescence intensity.
The invention also provides a preparation method of the near-infrared two-region fluorescent probe targeting the tyrosine kinase, which comprises the following steps of a) preparing the near-infrared two-region fluorescent probe targeting the tyrosine kinase by coupling a tyrosine kinase identification group TKP and a near-infrared two-region fluorescent group SQ1050 in a ratio of 1:1 through chemical bonds under an anhydrous condition; b) the near-infrared two-region fluorescent probe TKPSQ-2 targeting the tyrosine kinase is prepared by coupling the probe TKPSQ-1 and a tumor targeting polypeptide group PEP in a ratio of 1:1 through chemical bonds under anhydrous conditions.
The invention also provides a reagent for targeted tumor fluorescence imaging and dynamic tracking of tyrosine kinase, which contains the near-infrared two-region fluorescent probe of the targeted tyrosine kinase.
The invention also provides a reagent for tumor targeted near-infrared two-zone fluorescence imaging and tumor metastasis monitoring, which can be used as a reagent for surgical navigation.
The invention also provides a reagent for dynamically tracing the tumor tyrosine kinase, and the reagent contains the near-infrared two-region fluorescent probe of the targeted tyrosine kinase.
The invention also provides application of the near-infrared two-region fluorescent molecular probe of the targeted tyrosine kinase in preparing a reagent for tumor targeted near-infrared two-region fluorescent imaging and tumor metastasis monitoring.
The invention also provides application of the near-infrared two-region fluorescent molecular probe targeting the tyrosine kinase in preparing a reagent for monitoring the content and distribution of the tumor tyrosine kinase.
Detailed Description
The probe is formed by connecting a detection group and a fluorescent group through a chemical bond, is summarized as a structure I type, and can be represented as TKP-SQ 1050.
Specific examples of the near-infrared two-region fluorescent molecular probe targeting tyrosine kinase are as follows:
structure I type: TKP-SQ1050
The I-type structure is used as a near-infrared two-region fluorescent molecular probe of a targeted tyrosine kinase, molecular fluorescence is quenched due to aggregation-induced fluorescence quenching effect, and when the I-type structure is combined with the tyrosine kinase, the fluorescence intensity is greatly enhanced. The content of the tyrosine kinase in the tyrosine kinase solution and the cells is determined by quantitatively detecting the fluorescence intensity.
The technical scheme of the reagent serving as a near-infrared two-region fluorescent molecular probe of the targeted tyrosine kinase in tyrosine kinase solution and cells is as follows:
1) in common buffer solution such as PBS or Tris-HCl, the reaction temperature is between 20 ℃ and 45 ℃, and the optimal reaction time is preferably 37 ℃; the pH value of the incubation system is between 5.0 and 8.5, and the optimal reaction pH value is preferably pH 7.4; the reaction time is 5 to 30 minutes, preferably 20 minutes.
2) The content of the tyrosine kinase is evaluated by measuring the fold increase of the fluorescence intensity, and a fluorescence detector can be adopted to realize the rapid and sensitive detection of a product and a substrate; the fluorescence detection conditions were: the excitation wavelength is 950nm, and the fluorescence emission spectrum is detected at 1000-1100 nm.
Still another object of the present invention is to provide a reagent and a method for surgical navigation. The reagent can be prepared by connecting a type I reagent with a tumor targeting polypeptide group (PEP), can be used for real-time targeting fluorescence imaging of tumor cells or tissues, and can be summarized as a structure type II and can be expressed as TKP-SQ 1050-PEP.
Specific examples of agents that target fluorescence imaging tumor cells or tissues in real time are:
structure type II: TKP-SQ1050-PEP
The preparation method of the fluorescent probe comprises the following steps:
the preparation method of TKPSQ-1 comprises the following steps:
the preparation method of TKPSQ-2 comprises the following steps:
in organisms, the II type reagent is a target recognition group of tyrosine kinase and can be specifically targeted and gathered to tumor tissues. Therefore, the type II reagent can be applied to tumor boundary identification, residual tumor tissue identification and tumor metastasis lymph node tracking in the surgical tumor resection operation process, and a novel fluorescence navigation reagent is provided for a fluorescence navigation system of tumor surgery. After the reagent is injected into a living body with tumor diseases, tumor cells or tissues can emit fluorescence with a certain wavelength under the irradiation of light waves with specific wavelengths in the operation visual field of an organism, and doctors are guided to carry out surgical operations under an imaging system, so that the operation treatment effect is effectively improved, and the death rate of tumor patients is reduced.
The technical scheme of the application of the reagent serving as the surgical fluorescent navigation reagent in the tumor-bearing mouse is as follows:
1) the II type reagent can be dissolved in PBS solution or normal saline solution containing DMS 00% -5%, prepared into working solution, and injected into the body of a mouse by tail vein injection or local injection;
2) the type II reagent can be specifically gathered and targeted to a tumor part through blood circulation;
3) the type II reagent has no fluorescence signal or weak fluorescence signal intensity in a non-tumor tissue area, and has strong fluorescence signal intensity in a tumor tissue area.
4) The living body near-infrared two-zone fluorescence imaging technology is utilized, the tumor position is accurately imaged according to fluorescence signals, and navigation is provided for surgical operation.
The innovation of the invention is that 1, the specific combination of the tyrosine kinase recognition group and the tyrosine kinase is utilized to ensure that the probe has high specificity; 2. the probe has a reporter group SQ1050 which is an autonomous design and synthesis and is original. In addition, the near-infrared two-zone fluorescence imaging technology is a novel imaging technology which is emerging in recent years, and is far superior to the traditional visible light and near-infrared one-zone fluorescence imaging in the aspects of imaging sensitivity and resolution; 3. the tumor targeting group used by the II type reagent is a cyclic polypeptide which is designed and synthesized automatically, is designed and synthesized for the first time, and has targeting property on fibronectin on the surface of tumor cells, particularly breast cancer cells.
The invention has the beneficial technical effects that:
the invention provides a near-infrared two-region fluorescent probe targeting tyrosine kinase, which can be combined with tyrosine kinase in a biological system to enhance fluorescence, so that the probe can be used for detecting tyrosine kinase in a biological body; the probe can be specifically targeted to tumor tissues, and the fluorescence is enhanced after the probe is combined with tyrosine kinase, so that the probe can be used for tumor targeted fluorescence imaging and can also be used for dynamic tracing of the expression content and distribution of tyrosine kinase. Experiments show that the fluorescence intensity of the probe is weak, and the fluorescence is obviously enhanced after the probe is combined with tyrosine kinase.
In the invention, the change of the fluorescence signal after the tumor targeting agent is detected to identify the tumor is simple to operate and harmless to the body; the reagent realizes high sensitivity and high specificity in application, and provides an effective tool for tumor research in medicine, clinical monitoring and tumor metastasis treatment, so that the reagent has good application prospects in aspects of tumor targeted fluorescence imaging, dynamic tracing of tyrosine kinase expression content and distribution thereof and the like.
Drawings
FIG. 1 is a High Performance Liquid Chromatography (HPLC) and time-of-flight mass spectrometry analysis chart of the probe TKPSQ-1;
FIG. 2 is a High Performance Liquid Chromatography (HPLC) and time-of-flight mass spectrometry analysis chart of the probe TKPSQ-2;
FIG. 3 is a fluorescence spectrum before and after the reaction of the probe TKPSQ-1 and tyrosine kinase;
FIG. 4 is a kinetic experiment of the probe TKPSQ-1 with tyrosine kinase.
FIG. 5 shows fluorescence spectra of the probe TKPSQ-1 reacting with various interfering species and tyrosine kinase;
FIG. 6 shows fluorescence spectra of probe TKPSQ-2 reacted with various interfering species and tyrosine kinase;
FIG. 7 is fluorescent imaging of the probe TKPSQ-2 on a mouse model of breast cancer tumor bearing.
Detailed Description
For a better understanding of the present invention, the contents of the present invention will be further explained below with reference to the drawings and examples, but the contents of the present invention are not limited to the following examples.
The reagents used in the invention:
the instrument used in the invention:
| instrument for measuring the position of a moving object | Manufacturer of the product |
| High performance liquid chromatograph | GE,AKTA |
| Matrix-assisted laser desorption ionization time-of-flight mass spectrometer | ABSCIE,4800Plus |
| Fluorescence spectrometer | VarianCaryEclipse |
| Laser confocal microscope | Nikon |
| Small animal living body imaging system | PerkinElmer |
Example 1.
The synthesis process of the TKPSQ-1 probe comprises the following steps:
firstly, the synthetic steps of the self-designed and synthesized near-infrared two-region fluorescent group SQ1050 are shown as the following formula: weighing raw materials of 7.0mmol of compound 1 and 3.5mmol of compound 2, placing the raw materials into a solanaceous bottle, adding 50mL of n-butanol and toluene respectively, heating and refluxing for 8 hours, and rotatably evaporating the solvent to obtain an intermediate compound 3. Then, 3.5mmol of ninhydrin and 50mL of absolute ethanol are added into the intermediate compound 3, the mixture is heated and refluxed for 1 hour, the solvent is removed by rotary evaporation, and the mixture is recrystallized by methanol to obtain a near-infrared two-zone fluorescent group SQ1050, wherein the yield is 47.9%.
Finally, the synthesis procedure of TKPSQ-1 is shown as follows:
0.50mmol of the compound TKP, 0.50mmol of the compound SQ1050, 0.6mmol of DIEA, and 0.6mmol of EDC.HC1 were weighed into a 50mL three-necked flask, dissolved in 25mL of DMF under protection of argon, and stirred at room temperature for 10 hours. Vacuum distilling, removing solvent, and separating and purifying the crude product by high performance liquid chromatography to obtain TKPSQ-1 with yield of 92.5%, HPLC purity of 99%, MALDI-TOF (m/z): theoretical value 999.05, Experimental value [ M + H]+1000.04。
Example 2.
The synthesis process of the TKPSQ-2 probe comprises the following steps:
firstly, the synthetic steps of the autonomously designed cyclic polypeptide PEP are shown as the following formula:
(1) placing 2-chlorotrityl chloride resin (309mg,0.810mmol/g,0.25mmol,1equiv.) in a reaction flask, adding DCM for swelling, adding Fmoc-D-Glu (OBn) -OH (0.75mmol,3.0equiv) and DIEA (2.5mmol, 10equiv) for reacting for 3 hours to obtain resin coupled with D-Glu (OBn) -OH;
(2) adding organic base hexahydropiperidine, reacting in a DMF solution at room temperature for 3 hours, and removing the Fmoc protective group; Fmoc-Arg (Pbf) -OH (0.75mmol,3.0equiv), condensing agent DIC (1.5mmol, 6.0equiv) were added and Fmoc-Arg (Pbf) -OH was coupled to the resin;
(3) and (3) sequentially repeating the step (2), wherein the amino acids are sequentially replaced by Fmoc-Glu (OtBu) -OH, Fmoc-Lys (Boc) -OH and Fmoc-Ala-OH, and finally obtaining the resin A sequentially coupled with the amino acids.
(4) Adding organic base hexahydropiperidine, reacting in a DMF solution at room temperature for 3 hours, and removing the Fmoc protective group; resin A was cleaved with DCM containing 2.5% TFA and filtered with suction to give linear polypeptide B.
(5) And (3) placing the linear polypeptide B, 0.5mmol NHS and 0.5mmol EDC.HC1 into a 500mL three-necked flask, adding 250mL DMF for dissolving, stirring for 10 hours at room temperature under the protection of argon, coupling the first amino acid of the linear polypeptide B, and cyclizing to form the cyclic polypeptide C.
(6) And (2) placing the cyclic polypeptide C and 0.1mmol Pd/C catalyst in a 50mL three-necked flask, adding 25mL DMF for dissolving, introducing hydrogen, stirring at room temperature for 2 hours, removing a protecting group benzyl Bn, and separating by using a C18 preparation column to obtain the cyclic polypeptide PEP.
Finally, the synthesis procedure of TKPSQ-2 probe is shown as follows:
0.20mmol PEP polypeptide, 0.20mmol TKPSQ-1, 0.6mmol DIEA, 0.6mmol EDC.HC1 are weighed into a 50mL three-necked flask, dissolved in 25mL DMF under the protection of argon, and stirred at room temperature for 10 hours. Distilling under reduced pressure, removing solvent, adding 95% trifluoroacetic acid into the crude product, stirring at room temperature for 3 hours, distilling under reduced pressure, removing solvent to obtain black viscous crude product, and separating and purifying by high performance liquid chromatography to obtain the product TKPSQ-2 with yield of 84.2%. As shown in FIG. 2, HPLC purity was 99%, MALDI-TOF (m/z): theoretical value 1580.67, Experimental value [ M + H]+1581.65。
Example 3.
The change of the absorption spectrum and the fluorescence spectrum before and after the reaction of the probe TKPSQ-1 and the tyrosine kinase:
1mg of the probe TKPSQ-1 was weighed out and prepared into 1mL of 1mM dimethyl sulfoxide mother liquor. 25. mu.L of the above mother liquor was dropped into 0.5mL of phosphate buffer solution, 60. mu.L of 100. mu.g/mL tyrosine kinase solution was added, the volume was made to 1mL with phosphate buffer solution, and the reaction was carried out at 37 ℃ for 30min while measuring the fluorescence spectrum of the above solution every 5 min.
The results in FIG. 3 show that the fluorescence spectrum of the probe TKPSQ-1 in the invention is gradually enhanced at 1050nm with the addition of tyrosine kinase, and the fluorescence intensity is enhanced by about 10 times.
Example 4.
Fluorescence intensity curves of the probe TKPSQ-1 after reaction with tyrosine kinase at different concentrations:
25. mu.L of the above mother solution was dropped into 0.5mL of phosphate buffer solution, 0 to 100. mu.L of 100. mu.g/mL tyrosine kinase solution was added, the volume was then adjusted to 1mL using phosphate buffer solution, and after sufficient reaction at 37 ℃ for 30min, the fluorescence spectrum of the above solution was measured.
The results in FIG. 4 show that the fluorescence spectrum of the probe TKPSQ-1 in the invention is gradually enhanced at 1050nm with the addition of tyrosine kinase with different concentrations, and the fluorescence enhancement and the concentration of the tyrosine kinase are in a linear relationship in the range of 0-10 mug/mL, which indicates that the probe has high sensitivity to the tyrosine kinase.
Example 5.
Probe TKPSQ-1 reacts with other species in the organism (Selectivity study)
Adding 25 μ L of the above mother solution into 0.5mL of phosphate buffer solution, adding 10 μ g/mL tyrosine kinase or other species in organism, adding phosphate buffer solution to constant volume of 1mL, reacting at 37 deg.C for 30min, and measuring fluorescence spectrum of the above solution.
The results in FIG. 5 show that only tyrosine kinase can cause the probe TKPSQ-1 to generate obvious fluorescence enhancement, and the probe is proved to have high specificity to tyrosine kinase, while the existence of other species does not interfere with the determination of tyrosine kinase.
Example 6.
Probe TKPSQ-2 reacts with other species in the organism (Selectivity study)
1.6mg of the probe TKPSQ-2 was weighed out and prepared into 1mL of 1mM dimethyl sulfoxide mother liquor. Adding 25 μ L of the mother solution into 0.5mL of phosphate buffer solution, adding 10 μ g/mL of tyrosine kinase or other biomolecules, diluting to 1mL with phosphate buffer solution, reacting at 37 deg.C for 30min, and measuring fluorescence spectrum of the solution.
The results in FIG. 6 show that tyrosine kinase can cause the probe TKPSQ-2 to generate obvious fluorescence enhancement, and the addition of other biomolecules does not cause the probe TKPSQ-2 to generate fluorescence enhancement basically, which indicates that the probe has high specificity to tyrosine kinase.
Example 7.
The probe TKPSQ-2 is used for targeted imaging of tumors in a breast cancer tumor-bearing mouse model.
Firstly establishing a human breast cancer mouse model, culturing a large amount of human breast cancer MDA-MB-231 cells, collecting cells in logarithmic growth phase, removing culture solution, washing twice by PBS (phosphate buffer solution), and adjusting the cell density to lx 107In the case of the nude mice, 100. mu.L of cells were injected into the right lower limb, and then the nude mice were kept, and the growth of the nude mice was observed every day, and whether or not solid tumors had developed. After confirming that the mice are loaded with tumors, 100. mu.L of the probe TKPSQ-2 with a concentration of 1mM is injected into the tail vein after the tumor diameters reach 8-10mM, respectively. Fluorescence imaging was performed on mice at 0h, 0.5h, 1h, 2h, 4 h, and 6h after probe injection, and the fluorescence intensity of the tumors of the mice was observed and compared, and the results are shown in fig. 7. As can be seen from the fluorescence image data, the tumor tissue fluorescence intensity gradually increased. The result shows that the probe TKPSQ-2 can trace tyrosine kinase of tumor tissues on a living organism layer and can be used as a tumor surgery navigation reagent.
The technical scheme of the application of the probe as a surgical fluorescent navigation reagent in a tumor-bearing mouse is as follows:
1) the probe TKPSQ-2 can be dissolved in a PBS solution or a physiological saline solution to be prepared into a working solution, and is injected into a mouse body through tail vein injection or local injection;
2) the probe TKPSQ-2 can specifically aggregate and target to a tumor part through blood circulation;
3) the probe TKPSQ-2 has no fluorescent signal or weak fluorescent signal intensity in a non-tumor tissue area, and has strong fluorescent signal intensity in a tumor tissue area.
4) The living body near-infrared two-zone fluorescence imaging technology is utilized, the tumor position is accurately imaged according to fluorescence signals, and navigation is provided for surgical operation.
Claims (8)
1. The near-infrared two-region fluorescent probe of the targeted tyrosine kinase is characterized by having a structure of TKP-SQ1050 with a structure I or TKP-SQ1050-PEP with a structure II;
the structure of the TKP-SQ1050 with the structure I is shown as the following formula TKPSQ-1:
the structure of the TKP-SQ1050-PEP with the structure II is shown as the following formula TKPSQ-2:
2. the tyrosine kinase-targeted near-infrared two-region fluorescent probe of claim 1, which is characterized in that the probe does not have near-infrared two-region fluorescence, the probe formed by combining a tyrosine kinase recognition group in the probe and tyrosine kinase has strong near-infrared two-region fluorescence, and a fluorescence detector can be used for realizing rapid and sensitive detection of substrate fluorescence change.
3. The tyrosine kinase-targeted near-infrared two-region fluorescent probe according to claim 1, which is used as a ligand of tyrosine kinase to quantitatively determine the content of tyrosine kinase in different biological systems by quantitatively detecting the fluorescence intensity.
4. The method for preparing the tyrosine kinase-targeted near-infrared two-region fluorescent probe of claim 1, which is characterized in that: the TK PSQ-1 is prepared by coupling a single tyrosine kinase recognition group TKP on a near-infrared second-region fluorescent group SQ 1050; TK PSQ-2 is prepared by respectively coupling a tyrosine kinase recognition group TKP and a tumor targeting polypeptide group PEP on a near-infrared two-region fluorescent group SQ 1050:
the tyrosine kinase recognition group TKP has the structure that:
the structure of the near-infrared second-region fluorescent group SQ1050 is as follows:
the tumor targeting polypeptide group PEP has the structure as follows:
5. a reagent for targeted tumor fluorescence imaging and dynamic tracking of tyrosine kinase, which is characterized in that the reagent contains the tyrosine kinase-targeted near-infrared two-region fluorescent probe of claim 1.
6. Use of the tyrosine kinase-targeted near-infrared two-region fluorescent probe of claim 1 in the preparation of a surgical navigation reagent for tumor-targeted fluorescence imaging and dynamic tracking of tumor-targeted tyrosine kinases.
7. A reagent for dynamically tracking tumor tyrosine kinase, which is characterized in that the reagent contains the near-infrared two-region fluorescent probe targeting tyrosine kinase as claimed in claim 1.
8. Use of the tyrosine kinase-targeted near-infrared two-region fluorescent probe of claim 1 for preparing a reagent for dynamic tracking of tumor tyrosine kinases.
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