CN117164878A - Macromolecular contrast agent and preparation method and application thereof - Google Patents

Macromolecular contrast agent and preparation method and application thereof Download PDF

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CN117164878A
CN117164878A CN202311168436.7A CN202311168436A CN117164878A CN 117164878 A CN117164878 A CN 117164878A CN 202311168436 A CN202311168436 A CN 202311168436A CN 117164878 A CN117164878 A CN 117164878A
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peg
contrast agent
dtpa
pll
pei
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刘敏
李昕昕
刘广阔
刘小洁
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Jianghan University
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Jianghan University
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Abstract

The invention belongs to the field of materials, and relates to a macromolecular contrast agent, a preparation method and application thereof. The invention has the advantages that: (1) structural adjustability. The proportion of PEI to CD in the main structure PEI-CD can be regulated to regulate the content of CD, so that the number of sites capable of reacting with AD is regulated. The controllability of the ratio of the PEG to the PLL of the side chain structure adjusts the density of gadolinium and the size of the overall structure. (2) convenience of modification of targeting groups. The bio-orthogonal reaction is environment-friendly and has high reaction speed. The invention utilizes bio-orthogonal reaction to modify the targeting group on the polymer, and has simple reaction.

Description

Macromolecular contrast agent and preparation method and application thereof
Technical Field
The invention belongs to the field of materials, and relates to a macromolecular contrast agent, a preparation method and application thereof.
Background
In 2020, about 1930 ten thousand new cancer cases are worldwide, and 2840 ten thousand new cancer patients are expected to grow by 47% in 2040 as compared with 2020. Thus, cancer remains a major public health problem worldwide, severely threatening human health, and morbidity and mortality continue to rise. Early diagnosis and treatment would be an effective measure in reducing the incidence of cancer and mortality. MRI technology has become one of the most powerful detection means in clinical diagnosis at present due to the fact that the MRI technology has many outstanding advantages of no ionizing radiation, higher spatial resolution, higher contrast and the like. However, although the spatial resolution of MRI is high, the sensitivity of MRI imaging alone is not high. The overlapping of relaxation times at certain different tissues or tumor sites makes MRI challenging to discover early stage micro-tumors. There is thus a need to use contrast agents to assist imaging to enhance signal contrast and to increase resolution of tissue images. The contrast agent applied to clinic at present has the defects of low relaxation, short circulation time in blood and residence time in tissues, no targeting property and the like, and can not meet the requirement of early diagnosis of tumor.
Beta-cyclodextrin (beta-CD) is a cyclic sugar formed by connecting alpha-D type glucopyranose units end to end through alpha-1, 4 glycosidic bonds, and CD has a hydrophobic cavity and a hydrophilic surface, and can be used as a host molecule to be combined with inorganic, organic, biological and other guest molecules to form a saturated compound. Cyclodextrin has wide application in the fields of high molecular polymers, macromolecular self-assembly, biomedicine and the like. Macromolecular self-assembly refers to the process of spontaneously forming aggregates of a specific structure or morphology by the polymer under the pushing of non-covalent forces such as water transport, van der Waals forces, hydrogen bonds and the like. And cyclodextrin has low toxicity and good biocompatibility, so that the cyclodextrin is fully utilized in the aspect. Branched Polyethylenimine (PEI) is a dendrimer which has good water solubility, a narrow molecular weight distribution range, a single molecule, i.e. having nanometre dimensions, a multivalent effect, and easy modification of its physicochemical properties by modifying the surface groups. Can be applied to the preparation of macromolecular contrast agents. The macromolecular contrast agent not only can connect the targeting group and the micromolecular contrast agent at the same time and enhance the phagocytic capacity of specific cells so as to improve the specificity of the contrast, but also can connect various micromolecular contrast agents so as to perform various imaging and provide more abundant and accurate information for clinical diagnosis and treatment.
Disclosure of Invention
According to the defects of the prior art, the invention provides a macromolecular contrast agent, a preparation method and application thereof, cyclodextrin is built in a macromolecular host, a functional group is built on a side chain, and the contrast agent can be simply prepared through host-guest self-assembly, so that the macromolecular contrast agent is applied to tumor imaging technology.
The preparation method of the macromolecular contrast agent comprises the following steps:
(1) The beta-cyclodextrin reacts with branched PEI after being activated by CDI to obtain PEI-CD polymer;
(2) By N 3 -PEGn-NH 2 The terminal amine group initiates ring-opening polymerization of lysine NCA and is blocked by adamantyl chloride;
(3) Removing benzyl protection, and then modifying DTPA-Gd to polylysine to prepare a side chain N with MRI imaging capability 3 -PEGn-PLL(DTPA-Gd)-AD；
(4) PEI-CD is reacted with N by host guest interaction between cyclodextrin and adamantane 3 -PEG n -PLL (DTPA-Gd) -AD for host-guest interaction assembly to build macromolecular contrast agent;
(5) Passing the targeting group through DBCO and N 3 Is modified onto the synthesized polymer.
Wherein, as a preferable scheme, the mol ratio of the beta-cyclodextrin to the PEI in the step (1) is 1:1 to 6.
Preferably, N in step (2) 3 -PEG n -NH 2 The molecular weight of the PEG is in the range of n=400-20000.
Preferably, N in step (2) 3 -PEG n -NH 2 The molar ratio of terminal amine groups to lysine NCA is 1:20 to 70.
As a preferred embodiment, PEI-CD and N in step (4) 3 -PEG n -PLL (DTPA-Gd) -AD with a mass ratio of 1:10 to 20.
Preferably, the targeting group in step (5) is folic acid.
The invention also provides a macromolecular contrast agent which is prepared according to the method.
The invention also provides application of the macromolecular contrast agent in tumor imaging.
The invention has the advantages that:
(1) Adjustability of the structure. The proportion of PEI to CD in the main structure PEI-CD can be regulated to regulate the content of CD, so that the number of sites capable of reacting with AD is regulated. The controllability of the ratio of the PEG to the PLL of the side chain structure adjusts the density of gadolinium and the size of the overall structure.
(2) Convenience of targeting group modification. The bio-orthogonal reaction is environment-friendly and has high reaction speed. The invention utilizes bio-orthogonal reaction to modify the targeting group on the polymer, and has simple reaction.
Drawings
FIG. 1 is a reaction scheme of the present invention.
FIG. 2 is a PEI tested in example 6 800 -nuclear magnetic hydrogen profile of CyD.
FIG. 3 is a graph of N detected in example 6 3 -PEG 2000 -nuclear magnetic hydrogen profile of Lys (Z) -AD.
FIG. 4 is a graph of N detected in example 6 3 -PEG 2000 -PLL-AD and N 3 -PEG 2000 -nuclear magnetic hydrogen spectrogram of PLL (DTPA) -AD; wherein A is N 3 -PEG 2000 -PLL-AD, B is N 3 -PEG 2000 -PLL(DTPA)-AD。
FIG. 5 is a sample of PC/AD-PEG detected in example 7 2000 -PLL(DTPA-Gd)-N 3 Relaxation rate.
FIG. 6 is a sample of PC/AD-PEG detected in example 7 2000 -PLL(DTPA-Gd)-N 3 Solution imaging at different gadolinium concentrations.
FIG. 7 is a graph showing the analysis of cytotoxicity results of the macromolecular contrast agent tested in example 8.
FIG. 8 is a chart showing analysis of the results of the toxicity of the macromolecular contrast agent examined in example 9.
FIG. 9 is an in vivo Magnetic Resonance Imaging (MRI) diagram of the macromolecular contrast agent detected in example 10 in nude mice
FIG. 10 is a chart showing the MRI quantitative analysis of the macromolecular contrast agent detected in example 10 in nude mice.
FIG. 11 is a graph showing the residual tissue of the macromolecular contrast agent detected in example 11 in nude mice.
Detailed Description
The technical scheme of the invention will be further described in detail below with reference to specific embodiments. It is to be understood that the following examples are illustrative only and are not to be construed as limiting the scope of the invention. All techniques implemented based on the above description of the invention are intended to be included within the scope of the invention.
Unless otherwise indicated, the starting materials and reagents used in the following examples were either commercially available or may be prepared by known methods.
Example 1:
PEI 800 -preparation of CyD:
(1) Recrystallisation of beta-CD: the beta-CD was placed in a eggplant-shaped bottle, ultrapure water was added, and the oil bath was heated to 110℃until the beta-CD was completely dissolved. And naturally cooling to completely separate out the beta-CD, filtering to obtain a sample, repeating the operation for three times to obtain a recrystallized beta-CD crude product, and drying in vacuum at 60 ℃ to obtain the product.
(2) Activation of beta-CD: recrystallized beta-CD (2.1 g,1.85 mmol) and CDI (2.4 g,14.8 mmol) were dissolved in 10mL of super-dry DMF and N was introduced 2 And (5) protecting. After 1h of reaction at room temperature, the solid after precipitation was precipitated with cold diethyl ether, washed 3 times with diethyl ether and filtered and the precipitate was dissolved in ultra-dry DMSO.
(3)PEI 800 -synthesis of CyD: taking PEI 800 (2.9 g,3.63 mmol) is dissolved in ultra-dry DMSO, triethylamine (4.14 mL,29.6 mmol) is added, and the activated cyclodextrin DMSO solution in (2) is placed in an equilibration funnel and slowly added dropwise to the solution for at least 1.5h. After 12h of reaction, the product was stored at-20 ℃ after 3 days of filtration, lyophilization, dialysis against a dialysis bag with mw=14000.
Example 2:
synthesis of lysine NCA:
in a 250mL three-necked flask, H-Lys (Z) -OH (10 g,1.36 mol) was dispersed in 80mL dry THF and heated to 50deg.C. Triphosgene (4.8 g,16.17 mmol) was dissolved in 20mL of dry THF and placed in a constant pressure funnel to drop the solution. After a period of reaction, the solution gradually solidifies and the magnetons cannot be agitated. With the continued addition of triphosgene, the solid was gradually dissolved until a clear solution was formed. Then, N is introduced into 2 Half an hour and tail gas treatment with NaOH solution. Finally, the solution was concentrated and precipitated with petroleum ether, the precipitate was collected, dissolved with THF, reprecipitated, the operation was repeated 3 times, and filtered to obtain a white solid powder.
Example 3:
N 3 -PEG 2000 -PLL-AD synthesis:
(1) Synthesis of N 3 -PEG 2000 -Lys-PLL(Z)：N 3 -PEG 2000 -NH 2 (500 mg,0.25 mmol) was pumped into an oil pump and vacuum was applied to remove water from the feed by azeotropic distillation of toluene and water. Subsequently 10mL of ultra-dry DMF was added to complete dissolution, the prepared lysine NCA (3.03 g,9.8 mmol) was added to the solution, and the solution was heated in an oil bath at 40℃and N 2 The reaction was carried out for 48 hours under protection.
(2) Synthesis of N 3 -PEG 2000 -Lys (Z) -AD: adamantanoyl chloride (0.5 g,2.5 mmol) was added to the reaction and continued at N 2 The reaction is carried out for 24 hours under the heating of 40 ℃. After the reaction was completed, the reaction solution was precipitated with diethyl ether, and the precipitate obtained by centrifugation was washed with diethyl ether 3 times. The pale yellow product was obtained by vacuum drying.
(3) Deprotection synthesis of N 3 -PEG 2000 -PLL-AD：N 3 -PEG 2000 Lys (Z) -AD was dissolved in 20mL TFA, 5mL of 33wt.% hydrogen bromide was added. N-filled 2 And (3) protecting by a balloon, and reacting for 6 hours at room temperature to completely remove the protecting group from the product. The reaction solution was precipitated with diethyl ether and washed 3 times, dissolved with water, dialyzed (mw=3500) for 3 days, filtered and lyophilized to give a pale yellow product.
Example 4:
N 3 -PEG 2000 -synthesis of PLL (DTPA-Gd) -AD:
(1) Synthesis of DTPA-NHS (activationDTPA): diethylenetriamine pentaacetic acid (12 g,30.5 mmol) was dissolved in 220mL acetonitrile, 17mL triethylamine was added, and the mixture was taken up in N 2 Under protection, the oil bath is heated to 50 ℃ and reacts for 30min. After the whole reaction system was ice-bathed for 10min, N-hydroxysuccinimide (2.108 g,18.3 mmol) was dissolved in 50mL of acetonitrile, N, N-dicyclohexylcarbodiimide (3.781 g,18.3 mmol) was dissolved in 80mL of acetonitrile, and the mixture was placed in a constant pressure funnel, and the mixture was added dropwise to the reaction system, followed by reaction at room temperature for 8 hours after completion of the dropwise addition. After the reaction was completed, DCU was removed from the solution to give a pale yellow viscous solid, which was dissolved in 20mL of dry DMSO and stored at-20 ℃.
(2) Synthesis of N 3 -PEG 2000 -PLL (DTPA) -AD: will N 3 -PEG 2000 PLL-AD (0.3 g,0.045 mmol) was dissolved in 20mL dry DMSO, 0.68mL triethylamine was added, 5.35mL DTPA-NHS solution in DMSO was added, and after reaction for 12h. Then, 0.68mL of triethylamine and 5.35mL of DTPA-NHS were added thereto, and after the reaction for 12 hours, 0.68mL of triethylamine and 5.35mL of DTPA-NHS were added thereto again, and the reaction was continued for 12 hours. After the reaction was completed, the solution was dialyzed against a dialysis bag having mw=3500 for three days, and filtered and lyophilized to obtain a pale yellow solid.
(3) Synthesis of N 3 -PEG 2000 -PLL (DTPA-Gd) -AD: will N 3 -PEG 2000 -PLL (DTPA) -AD (1.4 g,0.15 mmol) dissolved in 20mL water, gdCl 3 ·6H 2 O was dissolved in 10mL of water and added to the above solution, and after adjusting the pH of the solution to 4-5 with 1M NaOH, the oil bath was heated to 40℃and reacted overnight. After the reaction was completed, the solution was dialyzed in an aqueous solution containing EDTA using a dialysis bag with mw=3500 for three days to remove unreacted Gd 3+ After further three days of dialysis in aqueous solution, the pale yellow product was obtained by lyophilization.
Example 5:
PC/AD-PEG 2000 -PLL(DTPA-Gd)-N 3 is synthesized by the following steps:
(1) Synthesis of DBCO-FA: folic acid (5.6 mg,0.0127 mmol) was dissolved in 5mL dry DMF, 0.007mL DIPEA and PyBOP (10.29 mg,0.019 mmol) were added, DBCO-PEG 2000 Amine (25.4 mg,0.0127 mmol) was dissolved in 2ml of LDMF and added to the above solution and reacted overnight. After the reaction was completed, the mixture was precipitated 3 times with diethyl etherAfter complete removal of the ether, it was dissolved in 100. Mu.L of water.
(2)PC/AD-PEG 2000- PLL(DTPA-Gd)-N 3 : PC (5 mg) was dissolved in 10mL of water, N 3 -PEG 2000 PLL (DTPA-Gd) -AD (60 mg) was dissolved in 10mL of water and added dropwise to the above solution, and after reacting for 12 hours, DBCO-FA (1 mg) was added and stirring was continued overnight. Finally, the solution was washed by centrifugation with an ultrafiltration centrifuge tube with mw=50000, unreacted starting material was removed, and concentrated to a yellowish solution.
Example 6:
nuclear magnetic resonance spectroscopy detection:
PEI testing Using Nuclear magnetic resonance spectrometer (400M) 800 -CyD、N 3 -PEG 2000 -Lys(Z)-AD、N 3 -PEG 2000 PLL-AD and N 3 -PEG 2000 -hydrogen profile of PLL (DTPA) -AD with the appropriate deuterated reagent as solvent and TMS as internal standard.
The nuclear magnetic results of PEI800-CyD are shown in FIG. 2. In the figure, δ4.92 (d, 1H) is shown, 4.03 to 2.84 are characteristic peaks of CyD, and δ2.14 to 2.81 are characteristic peaks of PEI. By comparing the peak area ratio of CyD to PEI, the ratio of beta-CD to PEI was calculated to be 1:2.3.
N 3 -PEG 2000 The nuclear magnetic results of-Lys (Z) -AD are shown in FIG. 3 by comparison with Arch 2 CH in O 2 (delta 4.96) and OCH in PEG 2 CH 2 The integrated area ratio of (δ3.5) gave an average degree of polymerization of 29.
N 3- PEG 2000 The nuclear magnetic results of PLL-AD are shown in FIG. 4, and after deprotection, the characteristic peaks of delta 7.43-6.89 benzene rings disappear, indicating complete removal of benzyl groups. N (N) 3 -PEG 2000 The nuclear magnetic results of the-PLL (DTPA) -AD are shown in fig. 3. The appearance of new peaks at delta 3.86ppm and delta 3.58-3.00 ppm, which are hydrogen on the methylene of DTPA, indicate that the DTPA has been successfully attached and the grafting ratio is calculated to be 96%.
Example 7:
relaxation rate measurement and T 1 Weighted solution imaging:
PC/AD-PEG 2000 -PLL(DTPA-Gd)-N 3 Solutions are configured to different gadolinium ion concentrationsGradient (0.25, 0.5,1,1.5,2 mM), 100. Mu.L each was measured for relaxation rates of different gadolinium ion concentrations using a pulsed nuclear magnetic resonance imager (0.5T). Relaxation rate r of sample 1 By 1/T 1 And (3) taking the gadolinium ion concentration as an abscissa and carrying out linear fitting to obtain the gadolinium ion concentration. In addition, sample solutions with different concentrations are prepared for T 1 Weighted imaging, T 1 The weighted imaging is to measure the transverse relaxation time of the material by using a spin echo sequence (CPMG) to obtain the recovery time (t) of the original data and the corresponding amplitude value M (t) . T1 weighted imaging uses spin echo sequences (te=8.6 ms, tr=100 ms) and the test temperature is kept at 35 ℃.
Measurement of PC/AD-PEG by reverse echo method using 0.5T pulse MRI 2000 -PLL(DTPA-Gd)-N 3 A slope several-bit relaxation rate r1 obtained by linear fitting with 1/T1 as ordinate and gadolinium ion concentration as abscissa. Relaxation rate of Gd-DTPA for clinical use is 4.25mM -1 ·S -1 The relaxation rate of the prepared contrast agent is 10.71mM as shown in FIG. 5 -1 ·S -1 Compared with the contrast agent which is superior to clinical application, the contrast agent is 2.58 times of that of the contrast agent. T1 weighted solution imaging is more capable of intuitively displaying the imaging effect of the prepared macromolecular contrast agent. As shown in fig. 6, the image brightness of the prepared contrast agent increases with an increase in gadolinium ion concentration.
Example 8:
cytotoxicity investigation:
the toxicity of the prepared contrast agent on 4T1 cells was determined by WST method. First, 4T1 cells were seeded into 96-well plates at 4000 cells per well, 100 μl of fresh DMEM medium was added to each well plate, and incubated in an incubator for 24 hours. PC/AD-PEG as contrast agent 2000 -PLL(DTPA-Gd)-N 3 The aqueous solution was diluted to different gadolinium ion concentrations (6, 3, 1.5, 0.75 mM). Materials of different Gd ion concentrations were added to the corresponding wells, 100 μl of each well, 100 μl of medium was added to the control group, and incubation was continued for 24h. Finally, the whole culture medium is sucked out by a pipette, 100 mu L of fresh culture medium and 10 mu L of CCK8 are added into each hole, and the mixture is placed in an incubator for cultivationCulturing for 2h, and measuring the absorbance at 450nm by using an enzyme-labeled instrument. 4 replicates were made for each gadolinium concentration and control group.
The relative viability of cells calculated using the formula is plotted on the ordinate and the gadolinium ion concentration on the abscissa. The results are shown in FIG. 7, PC/AD-PEG 2000 -PLL(DTPA-Gd)-N 3 Almost no toxicity to 4T1 cells, and when the gadolinium ion concentration reaches 6mM, the survival rate of U87 cells is kept above 85%, which indicates that the macromolecular gadolinium-based contrast agent has good biocompatibility.
Example 9:
tissue toxicity investigation:
by hematoxylin-eosin staining (H&E) To evaluate the safety of the prepared material and to examine the contrast agent PC/AD-PEG 2000 -PLL(DTPA-Gd)-N 3 Is a tissue toxicity of (a) a (b). 9 nude mice were taken, and were subjected to adaptive feeding with standard pellet diet and purified water in a constant temperature and constant humidity indoor environment for 1 week, periodically replaced with feed and sterilized. Nude mice were divided into three groups (n=3), two of which were intraperitoneally injected with 100 μl of PC/AD-PEG-containing mice, respectively 2000 -PLL(DTPA-Gd)-N 3 The amount of gadolinium ions was 0.1mmol/kg and 0.3mmol/kg, respectively. Another group of nude mice was used as a control group to which 100 μl of physiological saline was intraperitoneally injected. After 2 days of further feeding, nude mice were sacrificed by cervical dislocation, organs including heart, liver, spleen, lung and kidney were dissected and collected, immersed in formalin, stained sections were prepared, and placed under a microscope for histological analysis and photographed. Tissue H&The micrograph after E staining is shown in FIG. 8. Experimental results show that PC/AD-PEG injection was performed at 0.1mM/kg and 0.3mM/kg 2000 -PLL(DTPA-Gd)-N 3 No obvious damage to the tissue and organ of the mice, no necrosis in cardiac cells, no inflammatory reaction in liver cells, no sign of fibrosis in lung cells, and no morphological change in kidney cells. The organs of heart, liver, spleen, lung and kidney have almost no histological changes, and maintain the normal histological morphology. Therefore, PC/AD-PEG can be judged 2000 -PLL(DTPA-Gd)-N 3 Has good biocompatibility.
Example 10:
in vivo magnetic resonance imaging study:
the 9 tumor-bearing nude mice were divided into three groups, each of which was anesthetized by injecting 135 μl of a 25% uratam solution into the abdominal cavity, and the nude mice were fixed on a holder and subjected to blank scanning by a magnetic resonance imager of 4.7T, and the obtained pictures were used as blank references. The temperature was maintained at 35 ℃. Then, PC/AD-PEG was injected via tail vein, respectively 2000 -PLL(DTPA-Gd)-RGD，PC/AD-PEG 2000 -PLL(DTPA-Gd)-N 3 The concentration of gadolinium ions in the physiological saline solution of Gd-DTPA is 0.1mmol/kg for each group of samples. Following injection, nude mice were transferred to a 4.7T magnetic resonance imager for MRI imaging at the tumor site. The relative brightness of each group at each same time point was compared with the blank brightness of each group as 1.
As shown in fig. 9, it can be seen that the brightness of the tumor site in the targeted group was significantly higher than that in the non-targeted group with the increase of time, and in order to more intuitively compare the signal intensity enhancement effect of the tumor site, the Image J was processed to convert the brightness value of the tumor site before administration to 1 and the brightness of the tumor site after 1 hour and 2 hours to specific values, and the signal intensity increase factor was calculated, and the result is shown in fig. 10.
Example 11:
tissue residue:
taking 3 male nude mice (5 weeks old, about 20 g), and tail intravenous injection of 100 μL of prepared contrast agent PC/AD-PEG with gadolinium ion concentration of 0.1mmol/kg 2000 -PLL(DTPA-Gd)-N 3 Is a physiological saline solution of (a). After further feeding for 10 days in the SPF environment, nude mice were sacrificed by cervical dislocation, dissected and collected including heart, lung, liver, spleen, kidney, weighed and recorded sequentially. Then, about 100mg of each organ and tumor was placed in a 4mL centrifuge tube, 1mL nitric acid solution was added, and the mixture was heated to 60℃in a water bath for 1 hour. The solution was transferred to a 15mL centrifuge tube, and 10mL hydrogen peroxide was added and heated in a water bath at 90℃for 1 hour. Acid is removed after complete digestion by heating, and finally water is added for volume fixation to 60mL. 1mL of each sample was taken and the gadolinium content of the solution was determined by ICP-MS. Finally, the average gadolinium content in each organ is calculated and transformedReplaced by the percentage of Injected Dose (ID) per organ, tissue or tumor. The results are shown in FIG. 11. The residual amounts of gadolinium in heart, liver, spleen, lung and kidney were 0.01%, 0.28%, 0.03%, 0.02% and 0.02% per tissue or organ, respectively. These data demonstrate that the prepared macromolecular contrast agent has little long-term residue in major organs and tissues, and can initially meet the in-vivo long-term safety requirements of the contrast agent.

Claims (8)

1. The preparation method of the macromolecular contrast agent is characterized by comprising the following steps of:
(1) The beta-cyclodextrin reacts with branched PEI after being activated by CDI to obtain PEI-CD polymer;
(2) By N 3 -PEGn-NH 2 The terminal amine group initiates ring-opening polymerization of lysine NCA and is blocked by adamantyl chloride;
(3) Removing benzyl protection, and then modifying DTPA-Gd to polylysine to prepare a side chain N with MRI imaging capability 3 -PEGn-PLL(DTPA-Gd)-AD；
(4) PEI-CD is reacted with N by host guest interaction between cyclodextrin and adamantane 3 -PEG n -PLL (DTPA-Gd) -AD for host-guest interaction assembly to build macromolecular contrast agent;
(5) Passing the targeting group through DBCO and N 3 Is modified onto the synthesized polymer.
2. The method of claim 1, wherein the molar ratio of β -cyclodextrin to PEI in step (1) is 1:1 to 6.
3. The method of claim 1, wherein N in step (2) 3 -PEG n -NH 2 The molecular weight of the PEG is in the range of n=400-20000.
4. The method of claim 1, wherein N in step (2) 3 -PEG n -NH 2 The molar ratio of terminal amine groups to lysine NCA is 1:20 to 70.
5. The method of claim 1, wherein in step (4), PEI-CD and N are used as the main components 3 -PEG n -PLL (DTPA-Gd) -AD with a mass ratio of 1:10 to 20.
6. The method of claim 1, wherein the targeting group in step (5) is folic acid.
7. A macromolecular contrast agent prepared according to the method of claim 1.
8. Use of the macromolecular contrast agent of claim 7 in tumor imaging.
CN202311168436.7A 2023-09-12 2023-09-12 Macromolecular contrast agent and preparation method and application thereof Pending CN117164878A (en)

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