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A fluorescent probe for tumor-targeted diagnosis and treatment

A tumor targeting and probe technology, applied in the field of tumor targeted diagnosis and treatment fluorescent probes, can solve problems such as inability to feedback and monitor, and achieve the effects of improving solubility, simple purification process and simple preparation method.

Active Publication Date: 2018-07-27
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, individual differences and the complexity of the physiological environment may also cause false positive signals
Therefore, relying on a single change in fluorescence intensity cannot provide accurate feedback and monitoring of dynamic processes in living organisms

Method used

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  • A fluorescent probe for tumor-targeted diagnosis and treatment
  • A fluorescent probe for tumor-targeted diagnosis and treatment
  • A fluorescent probe for tumor-targeted diagnosis and treatment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] [Example 1] Synthesis of fluorescent probes for tumor-targeted diagnosis and treatment

[0049] Protoporphyrin-lysine (fluorescein)-serine-aspartic acid-glutamic acid-valine-aspartic acid-serine-lysine (dimethylaminoazobenzene)-arginine- Glycine-aspartic acid, PpIX-K(FAM)SDEVDSK(Dabcyl)RGD) synthesis at room temperature:

[0050] (1) Add 0.5 g of ammonia resin (0.525 mmol / g) to a reactor containing 10 mL of double-distilled N,N-dimethylformamide, and wait until the ammonia resin is dissolved in N,N-dimethylformamide N,N-dimethylformamide was extracted after swelling at room temperature for 2h.

[0051] (2) Add 20% (V / V) piperidine / N,N-dimethylformamide (i.e. the volume ratio of piperidine to N,N-dimethylformamide is 2:8) solution 10mL into the reactor After reacting at room temperature for 15 minutes, the solvent was removed; piperidine / N,N-dimethylformamide solution was repeatedly added for reaction to cut off the FMOC protecting group. After the reaction was complet...

Embodiment 2

[0063] [Example 2] Response detection of tumor-targeted diagnosis and treatment fluorescent probes to caspase-3

[0064] Dissolve the probe in HEPES buffer solution to make a working solution of 1 μmol / L. Apoptase-3 (1 U) was added to the probe-containing buffer solution, and the working solution was diluted with the buffer solution to a final concentration of 0.5 μmol / L. The fluorescence intensity of the luciferin in the solution was detected by a fluorescence spectrometer (LS55 fluorescence spectrophotometer, Perkin-Elmer) when the caspase-3 was just added and 11 hours after the caspase-3 was added. The excitation wavelength of fluorescein is 465 nm.

[0065] The result is as figure 2 As shown, the luciferin fluorescence intensity of the probe in the solution was weak when caspase-3 was just added, and after 11 hours of action with caspase-3, the luciferin intensity of the probe was about 11 times higher at 520 nm. enhanced. Therefore, it is proved that the fluorescence...

Embodiment 3

[0066] [Example 3] Specific detection of tumor-targeted diagnosis and treatment fluorescent probes responding to caspase-3

[0067] Apoptase-3 (1U) was incubated with a commercialized caspase-3 specific inhibitor (Ac-DEVD-CHO, 50 μmol / L) at 37°C for 2 hours. The probe solution was prepared as a 1 μmol / L working solution in HEPES buffer solution. Add apoptase-3 (1U) to the buffer solution containing the probe, apoptase-3 (1U) incubated with apoptase inhibitor, and dilute the probe concentration to a final concentration of 0.5 μmol / Lift. Use a fluorescence spectrometer to record how the fluorescence of the probe solution without adding captase-3, adding caspase-3, and adding caspase-3 and inhibitor changes with time. Excitation wavelength of fluorescein: 465 nm; emission wavelength of collected fluorescein: 520 nm.

[0068] The result is as image 3 As shown, without the action of apoptase-3, the fluorescein fluorescence intensity of the probe hardly recovers, while under t...

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Abstract

The invention discloses a tumor-targeted diagnosis and treatment integrated fluorescent probe which takes a tumor-targeted polypeptide sequence and an apoptosis enzyme specific recognition polypeptide sequence as a framework and comprises a fluorescence quenching molecular fluorescence pair and a photodynamic treatment photosensitizer. The diagnosis and treatment integrated fluorescent probe can achieve targeted treatment of tumors, and reduce toxic and side effects of the photodynamic treatment photosensitizer; a treatment result can be assessed in situ accurately in real time while the tumors are subjected to photodynamic treatment. The tumor-targeted diagnosis and treatment integrated fluorescent probe can also be used as a common fluorescent probe and used for screening of tumor treatment medicines and cell apoptosis imaging in a proportional fluorescence imaging manner. The tumor-targeted diagnosis and treatment integrated fluorescent probe can realize early detection of tumor treatment feedbacks and has great significance in promotion of accurate treatment of tumors and personal treatment.

Description

technical field [0001] The invention relates to a fluorescent probe for detecting tumor markers combined with photodynamic therapy to realize targeted therapy, a preparation method and application thereof. Background technique [0002] Cancer seriously threatens human health. Chemotherapy, radiotherapy and gene therapy have been widely used in cancer treatment research, but they are faced with the problem of how to further improve the effect of cancer treatment and reduce the side effects of drugs. The realization of targeted drug delivery to tumor sites has become an important issue in today's clinical medicine and biomedicine. One of the research hotspots in the field; the generation of multidrug resistance, changes in the tumor microenvironment, and individual differences lead to the failure of tumor treatment, realize personalized treatment methods for tumor treatment, timely feedback on tumor treatment effects, and quickly Optimizing the treatment plan will more effect...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C09K11/06C07K7/06A61K41/00G01N21/64
CPCA61K41/0061A61K41/0071C07K7/06C09K11/06C09K2211/1074C09K2211/1088G01N21/6486G01N2021/6417
Inventor 张先正李仕颖成红曾旋冯俊
Owner WUHAN UNIV
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