CN104069515B

CN104069515B - Paramagnetic metal complex modified by aspartic acid-leucine copolymer as well as preparation and application of paramagnetic metal complex

Info

Publication number: CN104069515B
Application number: CN201410300380.0A
Authority: CN
Inventors: 李晓晶; 肖研; 薛蓉; 湛游洋; 齐晨丽; 裴奉奎
Original assignee: Changchun Institute of Applied Chemistry of CAS
Current assignee: Changchun Institute of Applied Chemistry of CAS
Priority date: 2014-06-27
Filing date: 2014-06-27
Publication date: 2017-01-11
Anticipated expiration: 2034-06-27
Also published as: CN104069515A

Abstract

The invention relates to a paramagnetic metal complex modified by an aspartic acid-leucine copolymer as well as a preparation method and an application of the paramagnetic metal complex, and is used for solving the technical problems of low biodegradability, low liver targeting property, immunity defects and the like of the existing liver-type macromolecular contrast agent. The paramagnetic metal complex is formed by taking the aspartic acid-leucine copolymer as a carrier, wherein annular 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid ligands are connected on side chains of the carrier. A magnetic resonance imaging contrast agent prepared from the paramagnetic metal complex provided by the invention has the characteristics of a corresponding polyamine multi-carboxy complex, so that the magnetic resonance imaging contrast agent is good in stability, water solubility and relaxation rate, and simultaneously has the targeting property to the liver so as to realize the targeted imaging to improve the imaging contrast ratio and the imaging definition. Thus, the magnetic resonance imaging contrast agent has a good effect for improving the early-stage diagnosis level of diseases of the liver organs.

Description

The paramagnetic metal complex of aspartic acid-leucine copolymer modification and system thereof Preparation Method and application

Technical field

The present invention relates to a kind of magnetic resonance imaging contrast, be specifically related to a kind of aspartic acid-leucine copolymer and repair Paramagnetic metal complex of decorations and its preparation method and application.

Background technology

Compared with the little molecular contrast agents such as liver class macromolecular contrast agent and its spirits many, the former molecular dimension is relatively big, through hair Thin blood vessel speed is relatively slow, and the speed of rotation of molecule is relatively low, can improve relaxivity, reduces dosage, reduces toxicity.And by The equimolecular special biochemical property of polysaccharide, polypeptide, antibody, vitamin can make contrast agent be enriched in liver organization, thus realizes target To imaging, improve image contrast and definition.The macromolecular contrast agent of foreign study is by little molecular contrast agents Gd-mostly DOTA introduce high polymer main chain or covalent coupling to natural polymer, synthetic macromolecular carrier on formed, such as white egg -Gd-DOTA in vain, polyamide-Gd-DOTA etc., but, it is low that these liver class macromolecular contrast agents have biological degradability, liver Targeting is relatively low and the shortcoming such as immunity.

Summary of the invention

The invention solves the problems that in prior art that liver class macromolecular contrast agent has biological degradability low, liver targeting is relatively The technical problem of the shortcomings such as low and immunity, it is provided that the paramagnetic metal that a kind of aspartic acid-leucine copolymer is modified Coordination compound and its preparation method and application.

In order to solve above-mentioned technical problem, technical scheme is specific as follows:

The paramagnetic metal complex that a kind of aspartic acid-leucine copolymer is modified, the carrier of this coordination compound is sky Aspartic Acid-leucine copolymer, carrier side chain connects ring-type Isosorbide-5-Nitrae, 7,10-tetraazacyclododecanands-Isosorbide-5-Nitrae, and 7,10-tetra- The paramagnetic metal complex of acetic acid Hgand；Its structural formula is as follows:

Described aspartic acid-leucine copolymer is obtained by ASPARTIC ACID and L-Leu random copolymerization, Its molecular weight is 10000～400000；

The paramagnetic metal of described ring-type Cyclen-1,4,7,10-tetraacethyl part coordinates Thing is by 1:1 by Cyclen-1,4,7,10-tetraacethyl part (DOTA) and paramagnetic metal ion The paramagnetic metal complex that mol ratio coordination obtains；

A, b, c, d and n are natural number；

N^m+It is paramagnetic metal element+2 or+trivalent ion.

In technique scheme, described N^m+Be paramagnetic metal ion be Gd, Mn, Tb, Dy, Nd, Fe, Ti or Ru + 2 or+trivalent ion.

The preparation method of the paramagnetic metal complex that a kind of aspartic acid-leucine copolymer is modified, the method bag Include following steps:

(1) being mixed with L-Leu by ASPARTIC ACID, the mixture after mixing adds 8 grams～the mass fraction of 30 grams It is that the phosphoric acid solution of 85% stirs, reaction 1 hour～8 hours under conditions of pressure is 24mmHg～200mmHg, instead Answering temperature to control at 50 DEG C～220 DEG C, deionized water precipitates, and filters, washing, and drying under reduced pressure obtains aspartic acid-leucine Copolymer；Wherein mol ratio 10:1 of ASPARTIC ACID and L-Leu～1:9；Described aspartic acid-leucine copolymerization Thing structural formula is as follows:

Wherein, m and n is natural number；

(2) at room temperature aspartic acid-leucine copolymer synthesized in step (1) is dissolved in N, N-dimethyl methyl In amide solution, under stirring state, it is added dropwise over ethylenediamine, is stirred at room temperature 1 hour～5 hours, ether sedimentation, filter, dialysis, Lyophilizing, obtains aminated aspartic acid-leucine copolymer；Wherein ethylenediamine and aspartic acid-leucine copolymer Mass ratio be 2.8:1；Described aminated aspartic acid-leucine copolymer structure formula is as follows:

Wherein, a, b, c, d and n are natural number；

(3) at room temperature aminated aspartic acid-leucine copolymer synthesized in step (2) is dissolved in In ionized water, with potassium hydroxide solution regulation pH 7.0～9.0, the solution obtained is added drop-wise to DOTA mono-N-hydroxy succinum In acid imide active ester (Suflo-DOTA) solution, stirring 36 hours at room temperature, dialysis, concentrate, lyophilizing, obtain asparagine Cyclen-1,4,7,10-tetraacethyl the macromolecular ligand that acid-leucine copolymer is modified；Wherein amine The aspartic acid of base-leucine copolymer is 1:1 with the mass ratio of Suflo-DOTA；Described aspartic acid-leucine Cyclen-1,4,7,10-tetraacethyl macromolecular ligand the structural formula that copolymer is modified is as follows:

Wherein, a, b, c, d and n are natural number；

(4) the 1,4,7 at ambient temperature aspartic acid of synthesis-leucine copolymer in step (3) modified, 10-tetraazacyclododecanand-Isosorbide-5-Nitrae, 7,10-tetraacethyl macromolecular ligands and the reactant aqueous solution containing paramagnetic metal ion, PH value is adjusted to be 5.2 with potassium hydroxide solution, at room temperature stirring 1 day, dialysis, concentrate, lyophilizing, obtain aspartic acid-bright ammonia The paramagnetic metal complex that acid copolymer is modified；1,4,7,10-tetra-nitrogen that wherein aspartic acid-leucine copolymer is modified The material of the amount of the material of DOTA and paramagnetic metal ion in triazacyclododecane-1,4,7,10-tetraacethyl macromolecular ligand Amount ratio is 1:1.1；The structural formula of the paramagnetic metal complex that described aspartic acid-leucine copolymer is modified is as follows:

In technique scheme, the response time described in step (1) is 5h, and reaction temperature is 165 DEG C.

The paramagnetic metal complex modified with aspartic acid-leucine copolymer prepares magnetic resonance imaging contrast Method, the method comprises the following steps:

The paramagnetic metal complex that aspartic acid-leucine copolymer is modified is dissolved in sodium chloride injection, Portugal gathers In sugar injection, sodium chloride-dextran injection liquid or distilled water for injection, it is 6.5～8.0 with slow blood amine regulation pH, makes dense Degree is the magnetic resonance imaging contrast of 0.001～0.5 mol/L.

The paramagnetic metal complex that the aspartic acid of the present invention-leucine copolymer is modified can be by conventional method Make injection or injectable powder.Magnetic resonance imaging contrast prepared by the present invention can conventionally use.The magnetic of the present invention The dosage of resonance image-forming contrast agent can because of the kind of paramagnetic metal complex and as the diagnosis tissue of object or organ with And the difference of diagnostic device type and vary widely, typically, injection consumption is the diagnosis people of main body or other suckling is moved Every kg body weight 0.001 of object is to 5.0mmol, and preferably 0.05 arrives 0.5mmol.

Paramagnetic metal complex that the aspartic acid that the present invention provides-leucine copolymer is modified and preparation method thereof Provide the benefit that with application:

Magnetic resonance prepared by the paramagnetic metal complex that the aspartic acid that the present invention provides-leucine copolymer is modified Image-forming contrast medium, maintains the feature of corresponding polyamines many carboxylics coordination compound, thus has good stability, water solublity and relaxation Rate, has targeting simultaneously, thus realizes targeted imaging liver, improves image contrast and definition.To improving liver device The early diagnosis level of the disease of official has good result.Compared with its spirits many of wide clinical application, this kind of contrast agent has Advantages below:

(1) relaxivity is high, compared with its spirits many that clinic commonly uses, is about 2 times of its spirits many.Embodiment 1 is made Standby aspartic acid-leucine copolymer is by ethylene diamine-modified Cyclen-1,4,7,10- The relaxivity of tetraacethyl Gd coordination compound is up to 15.6mmol^-1·L·s^-1。

(2) imaging effect is good, and susceptiveness is high, and imaging time is long.Compared with its spirits many, injection targeting nuclear magnetic resonance is made After shadow agent, the picture signal of gained, definition and contrast also significantly improve.Its spirits many most at most can only maintain 30 minutes, And targeted contrast agent can make contrast agent be enriched in tissue or organ, the targeting contrast imaging time the longest (seeing accompanying drawing 1).

(3) there is good water solublity, it is easy to be configured to desired concn solution intravenous injection.

(4) aqueous stability is good, is suitable for pressure sintering sterilization.

(5) liver is had preferable selectivity: after intravenous injection is slightly below this type of contrast agent of clinical dosage, can be substantially Improve liver region image contrast (Wistar rat imaging experiment confirms, sees accompanying drawing 2).

(6) paramagnetic metal complex of this aspartic acid-leucine copolymer has preferably accumulation at liver (Kunming rat kidney tissue confirms, sees accompanying drawing 3).

(7) there is the concentration that the holding of the most relatively long time is stable, make liver obtain imaging window (ginseng steady in a long-term See accompanying drawing 3).

(8) liver to people or other mammal has good selectivity.

Animal imaging experiment: use Brooker company magnetic resonance imager, 30cm coil, 4.7T magnetic field, use T₁Weighting Multi-slice multi-echo imaging mode, repetition time TR:300ms, echo time TE:13.6ms, scanning area: 5.5 × 5.5cm², sweep Retouch matrix: 128 × 128.Take the Wistar rat of male Body Weight 190-210g, press 1.0mL/100g body weight fiber crops with 10% urethane After liquor-saturated, test axle position, animal abdominal cavity T₁Weighting picture, is prepared by 0.094mmol/kg body weight dose intravenous injection coordination compound of the present invention Magnetic resonance imaging contrast solution after imaging, every 3min sampling observation once, more than Continuous Observation 90min.Imaging results Showing, magnetic prepared by the paramagnetic metal complex that slightly below aspartic acid of clinical dosage-leucine copolymer is modified is altogether The image-forming contrast medium that shakes increases effect to liver magnetic resonance signal generation and is substantially better than the Gd-DOTA of clinical dosage, and whole reality The stage of testing is always maintained at good reinforced effects (seeing accompanying drawing 2).The raising of this contrast, shows this type of contrast agent good Good liver selective.

Accompanying drawing explanation

Fig. 1 is the time dependent enhancing of rat liver signal after magnetic resonance imaging contrast prepared by injection embodiment 28 Design sketch.

Fig. 2 be injection embodiment 28 preparation magnetic resonance imaging contrast before and injection 30min after rat axle position liver T₁ Weighting is as figure.

Fig. 3 is rat kidney tissue figure after magnetic resonance imaging contrast prepared by injection embodiment 28.

Detailed description of the invention

The invention thought of the present invention is: the present invention has biological fall from solving liver class macromolecular contrast agent prior art Solution is low, and liver targeting is relatively low and the technical problem of the shortcoming such as immunity is set out, and being prepared for one with copolymerization aminoacid is Carrier, carrier side chain connects the paramagnetism of ring-type DOTA part Metal complex.Copolymerization aminoacid is the class of amino acid polymeric derivative prepared based on amino acid polymorphisms.Copolymerization amino Acid has good biocompatibility and biological degradability, and the group of main chain both sides can provide medicine cross-linking agent for regulating its property The binding site of the suspension group of energy, as pharmaceutical carrier.Copolymerization aminoacid is the hydrolysis progressive cracking of acceptable enzyme in human body, its Catabolite is to human non-toxic's property.Using copolymerization aminoacid as the carrier of little molecular MRI contrast agent, form macromole bio-compatible Property copolymerization aminoacid MRI contrast agent, can reach for liver organ targeted imaging improve image contrast.

Embodiment 1

Aspartic acid-leucine copolymer passes through ethylene diamine-modified Cyclen-1,4,7, The preparation of 10-tetraacethyl Gd coordination compound

(1) by 20 grams of ASPARTIC ACIDs and 10 grams of L-Leu mixings, the mixture after mixing adds the quality of 10 grams Mark be 85% phosphoric acid solution in stir, decompression condition (24mmHg) under react 5 hours, reaction temperature controls 165 DEG C, deionized water precipitates, and filters, washing, and drying under reduced pressure obtains aspartic acid-leucine copolymer；

(2) at room temperature that aspartic acid-leucine copolymer is dissolved in N,N-dimethylformamide is molten by synthesized in (1) In liquid, being added dropwise over ethylenediamine under stirring state, ethylenediamine and aspartic acid-leucine copolymer quality are than for 2.8:1, room Temperature stirring 4 hours, ether sedimentation, filters, dialysis, lyophilizing, obtains aminated aspartic acid-leucine copolymer；

(3) at room temperature the aminated aspartic acid-leucine copolymer prepared in (2) is dissolved in deionized water In, it is 7.5 with potassium hydroxide solution regulation pH, the solution obtained is added drop-wise to DOTA mono-N-hydroxy thiosuccinimide active ester (Suflo-DOTA), in solution, aminated aspartic acid-leucine copolymer is 1:1 with the mass ratio of Suflo-DOTA, Stirring 36 hours at room temperature, dialysis, concentrate, lyophilizing, obtain the Isosorbide-5-Nitrae of aspartic acid-leucine copolymer, 7,10-tetra-nitrogen Triazacyclododecane-1,4,7,10-tetraacethyl macromolecular ligand；

(4) the 1,4,7,10-tetra-at ambient temperature aspartic acid-leucine copolymer obtained in (3) modified Azacyclo-dodecane-Isosorbide-5-Nitrae, 7,10-tetraacethyl macromolecular ligands and Gadolinium trichloride reactant aqueous solution, adjust pH value with potassium hydroxide solution It is 5.2, the thing of the amount of the material of DOTA and gadolinium ion in the DOTA macromolecular ligand that aspartic acid-leucine copolymer is modified The amount of matter ratio for 1:1.1, stirring 1 day at room temperature, dialysis, concentrate, lyophilizing, obtain aspartic acid-leucine copolymer and repair Cyclen-1,4,7,10-tetraacethyl the Gd coordination compound of decorations.

Embodiment 2

(1) by 2 grams of ASPARTIC ACIDs and 20 grams of L-Leu mixings, the mixture after mixing adds the quality of 8 grams and divides Number be 85% phosphoric acid solution in stir, decompression condition (30mmHg) under react 5 hours, reaction temperature controls 165 DEG C, deionized water precipitates, and filters, washing, and drying under reduced pressure obtains aspartic acid-leucine copolymer；

(3) at room temperature the aminated aspartic acid-leucine copolymer in (2) is dissolved in deionized water, Being 7.5 with potassium hydroxide solution regulation pH, the solution obtained is added drop-wise to DOTA mono-N-hydroxy thiosuccinimide active ester (Suflo-DOTA), in solution, aminated aspartic acid-leucine copolymer is 1:1 with the mass ratio of Suflo-DOTA, Stirring 36 hours at room temperature, dialysis, concentrate, lyophilizing, obtain the Isosorbide-5-Nitrae of aspartic acid-leucine copolymer, 7,10-tetra-nitrogen Triazacyclododecane-1,4,7,10-tetraacethyl macromolecular ligand；

Embodiment 3

(1) by 20 grams of ASPARTIC ACIDs and 20 grams of L-Leu mixings, the mixture after mixing adds the quality of 15 grams Mark be 85% phosphoric acid solution in stir, decompression condition (200mmHg) under react 5 hours, reaction temperature control At 100 DEG C, deionized water precipitates, and filters, washing, and drying under reduced pressure obtains aspartic acid-leucine copolymer；

Embodiment 4

(1) by 20 grams of ASPARTIC ACIDs and 20 grams of L-Leu mixings, the mixture after mixing adds the quality of 15 grams Mark be 85% phosphoric acid solution in stir, decompression condition (100mmHg) under react 5 hours, reaction temperature control At 50 DEG C, deionized water precipitates, and filters, washing, and drying under reduced pressure obtains aspartic acid-leucine copolymer；

Embodiment 5

(1) by 20 grams of ASPARTIC ACIDs and 40 grams of L-Leu mixings, the mixture after mixing adds the quality of 30 grams Mark be 85% phosphoric acid solution in stir, decompression condition (24mmHg) under react 5 hours, reaction temperature controls 165 DEG C, deionized water precipitates, and filters, washing, and drying under reduced pressure obtains aspartic acid-leucine copolymer；

Embodiment 6

(1) by 20 grams of ASPARTIC ACIDs and 80 grams of L-Leu mixings, the mixture after mixing adds the quality of 50 grams Mark be 85% phosphoric acid solution in stir, decompression condition (24mmHg) under react 4 hours, reaction temperature controls 200 DEG C, deionized water precipitates, and filters, washing, and drying under reduced pressure obtains aspartic acid-leucine copolymer；

Embodiment 7

(1) by 20 grams of ASPARTIC ACIDs and 40 grams of L-Leu mixings, the mixture after mixing adds the quality of 30 grams Mark be 85% phosphoric acid solution in stir, decompression condition (100mmHg) under react 1 hour, reaction temperature control At 220 DEG C, deionized water precipitates, and filters, washing, and drying under reduced pressure obtains aspartic acid-leucine copolymer；

Embodiment 8

(1) by 20 grams of ASPARTIC ACIDs and 5 grams of L-Leu mixings, the mixture after mixing adds the quality of 10 grams and divides Number be 85% phosphoric acid solution in stir, decompression condition (30mmHg) under react 5 hours, reaction temperature controls 200 DEG C, deionized water precipitates, and filters, washing, and drying under reduced pressure obtains aspartic acid-leucine copolymer；

Embodiment 9

Embodiment 10

(1) by 20 grams of ASPARTIC ACIDs and 2 grams of L-Leu mixings, the mixture after mixing adds the quality of 10 grams and divides Number be 85% phosphoric acid solution in stir, decompression condition (24mmHg) under react 8 hours, reaction temperature controls 200 DEG C, deionized water precipitates, and filters, washing, and drying under reduced pressure obtains aspartic acid-leucine copolymer；

Embodiment 11

(2) at room temperature that aspartic acid-leucine copolymer is dissolved in N,N-dimethylformamide is molten by synthesized in (1) In liquid, being added dropwise over ethylenediamine under stirring state, ethylenediamine and aspartic acid-leucine copolymer quality are than for 2.8:1, room Temperature stirring 1 hour, ether sedimentation, filters, dialysis, lyophilizing, obtains aminated aspartic acid-leucine copolymer；

(3) at room temperature the aminated aspartic acid-leucine copolymer in (2) is dissolved in deionized water, Being 7.0 with potassium hydroxide solution regulation pH, the solution obtained is added drop-wise to DOTA mono-N-hydroxy thiosuccinimide active ester (Suflo-DOTA), in solution, aminated aspartic acid-leucine copolymer is 1:1 with the mass ratio of Suflo-DOTA, Stirring 36 hours at room temperature, dialysis, concentrate, lyophilizing, obtain the Isosorbide-5-Nitrae of aspartic acid-leucine copolymer, 7,10-tetra-nitrogen Triazacyclododecane-1,4,7,10-tetraacethyl macromolecular ligand；

Embodiment 12

(2) at room temperature that aspartic acid-leucine copolymer is dissolved in N,N-dimethylformamide is molten by synthesized in (1) In liquid, being added dropwise over ethylenediamine under stirring state, ethylenediamine and aspartic acid-leucine copolymer quality are than for 2.8:1, room Temperature stirring 5 hours, ether sedimentation, filters, dialysis, lyophilizing, obtains aminated aspartic acid-leucine copolymer；

(3) at room temperature the aminated aspartic acid-leucine copolymer in (2) is dissolved in deionized water, Being 8.0 with potassium hydroxide solution regulation pH, the solution obtained is added drop-wise to DOTA mono-N-hydroxy thiosuccinimide active ester (Suflo-DOTA), in solution, aminated aspartic acid-leucine copolymer is 1:1 with the mass ratio of Suflo-DOTA, Stirring 36 hours at room temperature, dialysis, concentrate, lyophilizing, obtain the Isosorbide-5-Nitrae of aspartic acid-leucine copolymer, 7,10-tetra-nitrogen Triazacyclododecane-1,4,7,10-tetraacethyl macromolecular ligand；

Embodiment 13

(1) by 2 grams of ASPARTIC ACIDs and 20 grams of L-Leu mixings, the mixture after mixing adds the quality of 10 grams and divides Number be 85% phosphoric acid solution in stir, decompression condition (30mmHg) under react 5 hours, reaction temperature controls 165 DEG C, deionized water precipitates, and filters, washing, and drying under reduced pressure obtains aspartic acid-leucine copolymer；

Embodiment 14

(3) at room temperature the aminated aspartic acid-leucine copolymer in (2) is dissolved in deionized water, Being 9.0 with potassium hydroxide solution regulation pH, the solution obtained is added drop-wise to DOTA mono-N-hydroxy thiosuccinimide active ester (Suflo-DOTA), in solution, aminated aspartic acid-leucine copolymer is 1:1 with the mass ratio of Suflo-DOTA, Stirring 36 hours at room temperature, dialysis, concentrate, lyophilizing, obtain the Isosorbide-5-Nitrae of aspartic acid-leucine copolymer, 7,10-tetra-nitrogen Triazacyclododecane-1,4,7,10-tetraacethyl macromolecular ligand；

Embodiment 15

(2) at room temperature that aspartic acid-leucine copolymer is dissolved in N,N-dimethylformamide is molten by synthesized in (1) In liquid, being added dropwise over ethylenediamine under stirring state, ethylenediamine and aspartic acid-leucine copolymer quality are than for 2.8:1, room Temperature stirring 3 hours, ether sedimentation, filters, dialysis, lyophilizing, obtains aminated aspartic acid-leucine copolymer；

(3) at room temperature the aminated aspartic acid-leucine copolymer in (2) is dissolved in deionized water, Being 8.5 with potassium hydroxide solution regulation pH, the solution obtained is added drop-wise to DOTA mono-N-hydroxy thiosuccinimide active ester (Suflo-DOTA), in solution, aminated aspartic acid-leucine copolymer is 1:1 with the mass ratio of Suflo-DOTA, Stirring 36 hours at room temperature, dialysis, concentrate, lyophilizing, obtain the Isosorbide-5-Nitrae of aspartic acid-leucine copolymer, 7,10-tetra-nitrogen Triazacyclododecane-1,4,7,10-tetraacethyl macromolecular ligand；

Embodiment 16

Embodiment 17

Embodiment 18

Embodiment 19

Aspartic acid-leucine copolymer passes through ethylene diamine-modified Cyclen-1,4,7, The preparation of 10-tetraacethyl manganese complex

(4) the 1,4,7,10-tetra-at ambient temperature aspartic acid-leucine copolymer obtained in (3) modified Azacyclo-dodecane-Isosorbide-5-Nitrae, 7,10-tetraacethyl macromolecular ligands and manganese chloride reactant aqueous solution, adjust pH value with potassium hydroxide solution It is 5.2, the thing of the amount of the material of DOTA and manganese ion in the DOTA macromolecular ligand that aspartic acid-leucine copolymer is modified The amount of matter ratio for 1:1.1, stirring 1 day at room temperature, dialysis, concentrate, lyophilizing, obtain aspartic acid-leucine copolymer and repair Cyclen-1,4,7,10-tetraacethyl the manganese complex of decorations.

Embodiment 20

Embodiment 21

Embodiment 22

Aspartic acid-leucine copolymer passes through ethylene diamine-modified Cyclen-1,4,7, The preparation of 10-tetraacethyl terbium coordination compound

(4) the 1,4,7,10-tetra-at ambient temperature aspartic acid-leucine copolymer obtained in (3) modified Azacyclo-dodecane-Isosorbide-5-Nitrae, 7,10-tetraacethyl macromolecular ligands and terbium chloride reactant aqueous solution, adjust pH value with potassium hydroxide solution It is 5.2, the thing of the amount of the material of DOTA and terbium ion in the DOTA macromolecular ligand that aspartic acid-leucine copolymer is modified The amount of matter ratio for 1:1.1, stirring 1 day at room temperature, dialysis, concentrate, lyophilizing, obtain aspartic acid-leucine copolymer and repair Cyclen-1,4,7,10-tetraacethyl the terbium coordination compound of decorations.

Embodiment 23

Aspartic acid-leucine copolymer passes through ethylene diamine-modified Cyclen-1,4,7, The preparation of 10-tetraacethyl dysprosium coordination compound

(4) the 1,4,7,10-tetra-at ambient temperature aspartic acid-leucine copolymer obtained in (3) modified Azacyclo-dodecane-Isosorbide-5-Nitrae, 7,10-tetraacethyl macromolecular ligands and Dysprosium trichloride reactant aqueous solution, adjust pH value with potassium hydroxide solution It is 5.2, the thing of the amount of the material of DOTA and dysprosium ion in the DOTA macromolecular ligand that aspartic acid-leucine copolymer is modified The amount of matter ratio for 1:1.1, stirring 1 day at room temperature, dialysis, concentrate, lyophilizing, obtain aspartic acid-leucine copolymer and repair Cyclen-1,4,7,10-tetraacethyl dysprosium the coordination compound of decorations.

Embodiment 24

Aspartic acid-leucine copolymer passes through ethylene diamine-modified Cyclen-1,4,7, The preparation of 10-tetraacethyl Nd complex

(4) the 1,4,7,10-tetra-at ambient temperature aspartic acid-leucine copolymer obtained in (3) modified Azacyclo-dodecane-Isosorbide-5-Nitrae, 7,10-tetraacethyl macromolecular ligands and Neodymium chloride reactant aqueous solution, adjust pH value with potassium hydroxide solution It is 5.2, the thing of the amount of the material of DOTA and neodymium ion in the DOTA macromolecular ligand that aspartic acid-leucine copolymer is modified The amount of matter ratio for 1:1.1, stirring 1 day at room temperature, dialysis, concentrate, lyophilizing, obtain aspartic acid-leucine copolymer and repair Cyclen-1,4,7,10-tetraacethyl the Nd complex of decorations.

Embodiment 25

Aspartic acid-leucine copolymer passes through ethylene diamine-modified Cyclen-1,4,7, The preparation of 10-tetraacethyl iron complex

(4) the 1,4,7,10-tetra-at ambient temperature aspartic acid-leucine copolymer obtained in (3) modified Azacyclo-dodecane-Isosorbide-5-Nitrae, 7,10-tetraacethyl macromolecular ligands react with ferric chloride in aqueous solution, adjust pH value with potassium hydroxide solution It is 5.2, the thing of the amount of the material of DOTA and iron ion in the DOTA macromolecular ligand that aspartic acid-leucine copolymer is modified The amount of matter ratio for 1:1.1, stirring 1 day at room temperature, dialysis, concentrate, lyophilizing, obtain aspartic acid-leucine copolymer and repair Cyclen-1,4,7,10-tetraacethyl the iron complex of decorations.

Embodiment 26

Aspartic acid-leucine copolymer passes through ethylene diamine-modified Cyclen-1,4,7, The preparation of 10-tetraacethyl titanium complex

(4) the 1,4,7,10-tetra-at ambient temperature aspartic acid-leucine copolymer obtained in (3) modified Azacyclo-dodecane-Isosorbide-5-Nitrae, 7,10-tetraacethyl macromolecular ligands react with titanium chloride aqueous solution, adjust pH value with potassium hydroxide solution It is 5.2, the thing of the amount of the material of DOTA and titanium ion in the DOTA macromolecular ligand that aspartic acid-leucine copolymer is modified The amount of matter ratio for 1:1.1, stirring 1 day at room temperature, dialysis, concentrate, lyophilizing, obtain aspartic acid-leucine copolymer and repair Cyclen-1,4,7,10-tetraacethyl the titanium complex of decorations.

Embodiment 27

Aspartic acid-leucine copolymer passes through ethylene diamine-modified Cyclen-1,4,7, The preparation of 10-tetraacethyl ruthenium complex

(4) the 1,4,7,10-tetra-at ambient temperature aspartic acid-leucine copolymer obtained in (3) modified Azacyclo-dodecane-Isosorbide-5-Nitrae, 7,10-tetraacethyl macromolecular ligands react with ruthenium chloride aqueous solution, adjust pH value with potassium hydroxide solution It is 5.2, the thing of the amount of the material of DOTA and ruthenium ion in the DOTA macromolecular ligand that aspartic acid-leucine copolymer is modified The amount of matter ratio for 1:1.1, stirring 1 day at room temperature, dialysis, concentrate, lyophilizing, obtain aspartic acid-leucine copolymer and repair Cyclen-1,4,7,10-tetraacethyl the ruthenium complex of decorations.

Cyclen-1,4,7,10-the tetrem modified with aspartic acid-leucine copolymer It is as follows that acid paramagnetic metal complex prepares magnetic resonance imaging contrast method:

Embodiment 28

Take the 1,4,7,10-tetraazacyclododecane 12 that the aspartic acid-leucine copolymer of the embodiment 1 of 0.1 gram is modified Alkane-Isosorbide-5-Nitrae, 7,10-tetraacethyl Gd coordination compounds, it is dissolved in the sodium chloride injection of routine, is 6.5 with slow blood amine regulation pH value, system Become 0.001 mol/L magnetic resonance imaging contrast.

Accompanying drawing 1 is the time dependent increasing of rat liver signal after magnetic resonance imaging contrast prepared by injection the present embodiment Strong design sketch.This figure illustrates that magnetic resonance imaging contrast imaging effect prepared by coordination compound of the present invention is good, and susceptiveness is high, during imaging Between long.Compared with its spirits many, the picture signal of gained after injection targeting MRI contrast agents, definition and contrast are also Significantly improve.Its spirits many most at most can only maintain 30 minutes, and targeted contrast agent can make contrast agent be enriched in tissue or device In official, the targeting contrast imaging time is the longest.

Accompanying drawing 3 is rat kidney tissue figure after magnetic resonance imaging contrast prepared by injection the present embodiment.The explanation of this figure is originally Magnetic resonance imaging contrast prepared by invention coordination compound has preferably accumulation at liver, and has the most relatively long time Keep stable concentration, make liver obtain imaging window steady in a long-term.

Embodiment 29

Take the 1,4,7,10-tetraazacyclododecane 12 that the aspartic acid-leucine copolymer of the embodiment 5 of 0.1 gram is modified Alkane-Isosorbide-5-Nitrae, 7,10-tetraacethyl Gd coordination compounds are dissolved in the dextran injection liquid of routine, are 8.0 with slow blood amine regulation pH value, make 0.01 mol/L magnetic resonance imaging contrast.

Embodiment 30

Take the 1,4,7,10-tetraazacyclododecane 12 that the aspartic acid-leucine copolymer of the embodiment 10 of 0.1 gram is modified Alkane-Isosorbide-5-Nitrae, 7,10-tetraacethyl Gd coordination compounds are dissolved in the sodium chloride-dextran injection liquid of routine, with slow blood amine regulation pH value are 6.5, make 0.1 mol/L magnetic resonance imaging contrast.

Embodiment 31

Take the 1,4,7,10-tetraazacyclododecane 12 that the aspartic acid-leucine copolymer of the embodiment 15 of 0.1 gram is modified Alkane-Isosorbide-5-Nitrae, 7,10-tetraacethyl Gd coordination compounds are dissolved in distilled water for injection, are 8.0 with slow blood amine regulation pH value, make 0.5 and rub That/liter magnetic resonance imaging contrast.

Embodiment 32

Take the 1,4,7,10-tetraazacyclododecane 12 that the aspartic acid-leucine copolymer of the embodiment 19 of 0.1 gram is modified Alkane-Isosorbide-5-Nitrae, 7,10-tetraacethyl manganese complexes are dissolved in the dextran injection liquid of routine, are 8.0 with slow blood amine regulation pH value, make 0.01 mol/L magnetic resonance imaging contrast.

Embodiment 33

Take the 1,4,7,10-tetraazacyclododecane 12 that the aspartic acid-leucine copolymer of the embodiment 20 of 0.1 gram is modified Alkane-Isosorbide-5-Nitrae, 7,10-tetraacethyl manganese complexes are dissolved in the sodium chloride-dextran injection liquid of routine, with slow blood amine regulation pH value are 6.5, make 0.1 mol/L magnetic resonance imaging contrast.

Embodiment 34

Take the 1,4,7,10-tetraazacyclododecane 12 that the aspartic acid-leucine copolymer of the embodiment 8 of 0.1 gram is modified Alkane-Isosorbide-5-Nitrae, 7,10-tetraacethyl Gd coordination compounds are dissolved in the sodium chloride-dextran injection liquid of routine, with slow blood amine regulation pH value are 6.5, make 0.1 mol/L magnetic resonance imaging contrast.

Embodiment 35

Take the Wistar rat of male 100-120g body weight, after pressing the anesthesia of 1.0mL/100g body weight with 10% urethane, press After the magnetic resonance imaging contrast of 0.1mmol/kg body weight dose intravenous injection embodiment 28 preparation, test axle position, animal abdominal cavity T₁ Weighting picture, every 10min sampling observation once, more than Continuous Observation 120min.Obtain the liver of this magnetic resonance imaging contrast Axle position T₁Weighting picture, such as accompanying drawing 2.

Accompanying drawing 2 imaging results shows, the paramagnetism that slightly below aspartic acid of clinical dosage-leucine copolymer is modified Magnetic resonance imaging contrast prepared by metal complex is substantially better than clinical agent to the increase effect that liver magnetic resonance signal produces Amount Gd-DOTA, and the whole experimental stage be always maintained at good reinforced effects.The raising of this contrast, shows this The liver selective that class contrast agent is good.

Obviously, above-described embodiment is only for clearly demonstrating example, and not restriction to embodiment.Right For those of ordinary skill in the field, can also make on the basis of the above description other multi-form change or Variation.Here without also cannot all of embodiment be given exhaustive.And the obvious change thus extended out or Change among still in the protection domain of the invention.

Claims

1. the paramagnetic metal complex that aspartic acid-leucine copolymer is modified, it is characterised in that this coordination compound Carrier is aspartic acid-leucine copolymer, and carrier side chain connects ring-type Isosorbide-5-Nitrae, 7,10-tetraazacyclododecanand-1, The paramagnetic metal complex of 4,7,10-tetraacethyl part；Its structural formula is as follows:

Described aspartic acid-leucine copolymer is obtained by ASPARTIC ACID and L-Leu random copolymerization, its point Son amount is 10000～400000；

The paramagnetic metal complex of described ring-type Cyclen-1,4,7,10-tetraacethyl part is 1:1 mole is pressed by Cyclen-1,4,7,10-tetraacethyl part (DOTA) and paramagnetic metal ion The paramagnetic metal complex obtained than coordination；

A, b, c, d and n are natural number；

N^m+It is paramagnetic metal element+2 or+trivalent ion.

The paramagnetic metal complex that aspartic acid the most according to claim 1-leucine copolymer is modified, its feature It is, described N^m+Be paramagnetic metal ion be Gd, Mn, Tb, Dy, Nd, Fe, Ti or Ru+2 or+trivalent ion.

The system of the paramagnetic metal complex that aspartic acid the most according to claim 1 and 2-leucine copolymer is modified Preparation Method, it is characterised in that the method comprises the following steps:

(1) being mixed with L-Leu by ASPARTIC ACID, the mixture after mixing adds the mass fraction of 8 grams～30 grams and is The phosphoric acid solution of 85% stirs, reacts 1 hour～8 hours under conditions of pressure is 24mmHg～200mmHg, reaction Temperature controls at 50 DEG C～220 DEG C, and deionized water precipitates, and filters, washing, and drying under reduced pressure obtains aspartic acid-leucine common Polymers；Wherein mol ratio 10:1 of ASPARTIC ACID and L-Leu～1:9；Described aspartic acid-leucine copolymer Structural formula is as follows:

Wherein, m and n is natural number；

(2) at room temperature aspartic acid-leucine copolymer synthesized in step (1) is dissolved in N,N-dimethylformamide In solution, under stirring state, it is added dropwise over ethylenediamine, is stirred at room temperature 1 hour～5 hours, ether sedimentation, filter, dialysis, lyophilizing, Obtain aminated aspartic acid-leucine copolymer；Wherein ethylenediamine and the quality of aspartic acid-leucine copolymer Ratio is 2.8:1；Described aminated aspartic acid-leucine copolymer structure formula is as follows:

Wherein, a, b, c, d and n are natural number；

(3) at room temperature aminated aspartic acid-leucine copolymer synthesized in step (2) is dissolved in deionization In water, with potassium hydroxide solution regulation pH 7.0～9.0, it is sub-that the solution obtained is added drop-wise to DOTA mono-N-hydroxy succinyl In amine active ester (Suflo-DOTA) solution, at room temperature stirring 36 hours, dialysis, concentrate, lyophilizing, obtain aspartic acid- Cyclen-1,4,7,10-tetraacethyl the macromolecular ligand that leucine copolymer is modified；Wherein amido Aspartic acid-leucine the copolymer changed is 1:1 with the mass ratio of Suflo-DOTA；Described aspartic acid-leucine is common Cyclen-1,4,7,10-tetraacethyl macromolecular ligand the structural formula that polymers is modified is as follows:

Wherein, a, b, c, d and n are natural number；

(4) the 1,4,7,10-tetra-at ambient temperature aspartic acid of synthesis-leucine copolymer in step (3) modified Azacyclo-dodecane-Isosorbide-5-Nitrae, 7,10-tetraacethyl macromolecular ligands and the reactant aqueous solution containing paramagnetic metal ion, use hydrogen-oxygen Changing potassium solution adjusts pH value to be 5.2, stirring 1 day at room temperature, and dialysis concentrates, lyophilizing, obtains aspartic acid-leucine copolymerization The paramagnetic metal complex that thing is modified；The 1,4,7,10-tetraazacyclododecane ten that wherein aspartic acid-leucine copolymer is modified In dioxane-1,4,7,10-tetraacethyl macromolecular ligand, the amount of the material of DOTA with the amount ratio of the material of paramagnetic metal ion is 1:1.1；The structural formula of the paramagnetic metal complex that described aspartic acid-leucine copolymer is modified is as follows:

The preparation of the paramagnetic metal complex that aspartic acid the most according to claim 3-leucine copolymer is modified Method, it is characterised in that the response time described in step (1) is 5h, reaction temperature is 165 DEG C.

5. the paramagnetic metal complex modified with the aspartic acid described in claim 1 or 2-leucine copolymer prepares magnetic The method of resonance image-forming contrast agent, it is characterised in that the method comprises the following steps:

The paramagnetic metal complex that aspartic acid-leucine copolymer is modified is dissolved in sodium chloride injection, glucosan note Penetrating in liquid, sodium chloride-dextran injection liquid or distilled water for injection, be 6.5～8.0 with slow blood amine regulation pH, making concentration is The magnetic resonance imaging contrast of 0.001～0.5 mol/L.