Polypeptide molecule image probe targeting mesenchymal stem cell, preparation method of polypeptide molecule image probe, and mesenchymal stem cell marked by polypeptide molecule image probe

A kind of stem cell and molecular imaging technology, applied in the field of medical imaging, can solve the problems of low relaxation rate and large dose, achieve high affinity and target specificity, reduce safety problems, and enhance the effect of magnetic resonance weighted imaging contrast

Inactive Publication Date: 2016-07-06
SUZHOU INST OF NANO TECH & NANO BIONICS CHINESE ACEDEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, a key problem faced by Gd complex contrast agents is that their relaxation rates are much lower than T 2 The relaxation rate of superparamagnetic iron oxide nanoparticles, such as the longitudinal relaxation rate r of commercial small molecule contrast agents Gd-DTPA and Gd-DOTA 1 Usually 3-5mM -1 the s -1 , while the

Method used

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  • Polypeptide molecule image probe targeting mesenchymal stem cell, preparation method of polypeptide molecule image probe, and mesenchymal stem cell marked by polypeptide molecule image probe
  • Polypeptide molecule image probe targeting mesenchymal stem cell, preparation method of polypeptide molecule image probe, and mesenchymal stem cell marked by polypeptide molecule image probe
  • Polypeptide molecule image probe targeting mesenchymal stem cell, preparation method of polypeptide molecule image probe, and mesenchymal stem cell marked by polypeptide molecule image probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: (Gd-DOTA) 2-Synthesis of EM7 molecular imaging probe

[0042] 1. Bu t 3 Synthesis of DOTA (1,4,7-tris(tert-butoxycarbonylmethyl)-10-(acetic acid)-1,4,7,10-tetraazacyclododecane). The synthesis proceeds from 1,4,7,10-tetraazacyclododecane, a macrocyclic polyamine, as follows:

[0043] a) Weigh out 10.0g 1,4,7,10-tetraazacyclododecane and 29.3g NaHCO 3 In a one-liter three-necked flask, add 50 mL of acetonitrile. Weigh 37.4 g of tert-butyl bromoacetate in a fume hood, add 20 mL of acetonitrile, mix well, and put it into a dropping funnel. A solution of t-butyl bromoacetate in acetonitrile was slowly added dropwise to the reaction mixture under an ice bath and nitrogen protection. After the dropwise addition was completed, stirring was continued at room temperature for 30 hours. The solid was filtered off, the acetonitrile was removed by rotary evaporation, and recrystallized twice with toluene to obtain a white solid Bu t 3 DO3A (1,4,7-tris(tert-butoxy...

Embodiment 2

[0050] Example 2: (Gd-DOTA) 2 - Synthesis of CC9 Molecular Imaging Probe

[0051] 1. Synthetic Bu t 3 DOTA: 1,4,7-tris(tert-butoxycarbonylmethyl)-10-(acetic acid)-1,4,7,10-tetraazacyclododecane. The synthesis proceeds from 1,4,7,10-tetraazacyclododecane as follows:

[0052] a) Synthesize Bu according to Example 1 t 3 DO3A.

[0053] b) Weigh 1.38gK 2 CO 3 and 2.57gBu t 3 DO3A in a 250mL three-necked flask, add 50mL acetonitrile, N 2 Protected and stirred at room temperature, 1.37 g of benzyl bromoacetate in 5 mL of acetonitrile was added dropwise, and the reaction was carried out at 70 °C for 12-24 hours. Cool to room temperature, remove the solids by filtration, and remove the solvent by rotary evaporation. with CH 2 Cl 2 / MeOH (20:1) was used as the developing solvent for column chromatography separation, and the product 1,4,7-tris(tert-butoxycarbonylmethyl)-10-(benzyloxycarbonylmethyl) was obtained as pale yellow viscous foamy product )1,4,7,10-tetraazacyclodo...

Embodiment 3

[0059] Example 3: Preparation of Mesenchymal Stem Cells Labeled with Polypeptide Molecular Imaging Probes

[0060] 1. The polypeptide molecular imaging probe prepared in Example 1 or 2 was dissolved in the culture medium to form a concentration series of 0.05, 0.1, 0.2, 0.5, 1.0, 2.0 mM, and the mesenchymal stem cells were cultured in a D=90 mm dish 24 hours.

[0061] 2. Cell counting and cell dense packing: remove the medium, wash with PBS (2 mL×3) at pH 7.4, and remove unbound molecular imaging probes. Add 1.0 mL of trypsin for digestion, add 3 mL of PBS, and blow off the cells. Take 20 μL on a cell counting plate and count the number of cells under a microscope. All the remaining cells were transferred to a 10 mL centrifuge tube, and the culture dish was washed with 3 mL of PBS, and also transferred to the centrifuge tube. PBS was removed by centrifugation at 1200 rpm for 5 minutes. The cells were transferred to a capillary with an inner diameter of 1.5 mm and a capilla...

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Abstract

The invention discloses a polypeptide molecular imaging probe targeting mesenchymal stem cells, a preparation method thereof and mesenchymal stem cells marked by the probe. The molecular probe includes: a polypeptide for identifying mesenchymal stem cells and Contrast unit metal Gd complexes for enhanced contrast in magnetic resonance imaging. A main feature of this molecular probe is that a polypeptide probe binds 1 to 4 metal Gd complex molecules through dendrimers, and spacers and linkers are used to improve the complexation of polypeptides with dendrimers and dendrimers with Gd space between objects. The polypeptide molecular imaging probe can significantly enhance the contrast of magnetic resonance weighted imaging.

Description

technical field [0001] The invention relates to the field of medical imaging, in particular to a polypeptide molecular imaging probe targeting mesenchymal stem cells, which can enhance the contrast of magnetic resonance weighted imaging. Background technique [0002] Stem cells are a kind of cell population with self-renewal ability and multi-directional differentiation potential. They can maintain the number of their own cell groups through symmetrical division, and can also further differentiate into different tissue cells, thus forming various complex tissues and organs of the body. . Self-renewal and differentiation are the two main characteristics of stem cells. The basic idea of ​​stem cell regenerative medicine is to achieve regeneration and repair of damaged tissues and organs by inducing the directional differentiation of stem cells in the transplanted body. In stem cell research, real-time tracking of stem cell survival, migration, homing, directional differentia...

Claims

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Application Information

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IPC IPC(8): A61K49/14A61K49/08
Inventor 邓宗武谭波李彬彬张宏岩张海禄
Owner SUZHOU INST OF NANO TECH & NANO BIONICS CHINESE ACEDEMY OF SCI
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