A radionuclide-labeled polymer, its preparation method and application

Abstract

The present invention discloses a radionuclide-labeled polymer, a preparation method thereof and an application. Specifically, the present invention provides a radionuclide-labeled polymer having the following structure: G-L1-L2-P; wherein, G is a group containing a radionuclide, which includes a chelating group and a radionuclide, the chelating group chelates with the radionuclide, L1 and L2 are independently a chemical bond or a linking group; P is a temperature-sensitive structural unit. The radionuclide-labeled polymer of the present invention can be used as a tracer for hydrogels, and can have good compatibility and stability when combined with hydrogels without changing the properties of the hydrogels themselves. Further, it can be used for the tracing of in-vivo implantation of hydrogels to continuously and real-time track the position and state of the hydrogels after in-vivo implantation.

Description

Technical Field

[0001] The present invention relates to a radionuclide-labeled polymer, a preparation method thereof, and an application thereof. Background Art

[0002] With the rapid development of biomedical technologies, biocompatible hydrogels have been widely used in multiple biomedical fields such as filling, repair, drug delivery, interventional embolization, etc. For example, hydrogels have been used as advanced materials for vascular embolization in the treatment of tumor diseases with rich blood supply, such as hepatocellular carcinoma.

[0003] There is no method to continuously and real-time track the state and position of hydrogels after in vivo implantation, which causes great difficulties in evaluating the in vivo performance of implantable hydrogels. Although existing angiography can image the embolization of blood vessels to a certain extent, the imaging duration is limited, and it cannot continuously observe the state and position of implantable hydrogels, and cannot meet the imaging requirements of hydrogel materials with a long implantation period. Summary of the Invention

[0004] Aiming at the problem that the existing technology cannot meet the imaging requirements of hydrogel materials with a long implantation period, the present invention provides a radionuclide-labeled polymer, a preparation method thereof, and an application thereof. The radionuclide-labeled polymer provided by the present invention can be used as a tracer for hydrogels, does not change the properties of the hydrogels themselves, has good compatibility with hydrogels, good binding stability, and can maintain a high radioactive purity both in vivo and in vitro for more than 11 days. It can be used as a tracer for in vivo implantation of hydrogels to continuously and real-time track the position and state of hydrogels after in vivo implantation, and can be used for evaluating the in vivo performance of implantable hydrogels.

[0005] The present invention provides a radionuclide-labeled polymer having the following structure: G-L 1 -L 2 -P;

[0006] Wherein, G is a group chelating a radionuclide, which includes a chelating group and a radionuclide, and the chelating group chelates with the radionuclide;

[0007] L 1 and L 2 are independently a chemical bond or a linking group;

[0008] P is a temperature-sensitive structural unit.

[0009] In one embodiment, G consists of a chelating group and a radionuclide.

[0010] In one embodiment, the radionuclide is 89 Zr;

[0011] The chelating group is a chelating group formed by a bifunctional chelating agent.

[0012] In one embodiment, the L 1 is a chemical bond or a linking group represented by formula (I-1):

[0013] (I-1)

[0014] In formula (I-1), x and y are independently selected from the integers 0-3;

[0015] R 1 is , an alkylene group of C 1-5 , or a chemical bond;

[0016] R e is , a substituted or unsubstituted arylene group of C 6-10 , or, a substituted or unsubstituted heteroarylene group of C 5-10 ;

[0017] R d is an alkylene group of C 1-5 , or a chemical bond;

[0018] R b is hydrogen, a hydroxyl group or an alkyl group of C 1-5 ;

[0019] X is O or S; e represents the connection site of G and L 1 , i represents the connection site of R 1 and R e , f represents the connection site of L 1 and L 2 ;

[0020] The L 2 is a chemical bond, a group represented by formula (II-1) or (II-2):

[0021] (II-1)

[0022] (II-2),

[0023] wherein, in formula (II-1), z is 0, 1, 2 or 3;

[0024] In formula (II-2), R 2 and R 3 are independently hydrogen, an alkyl group of C 1-5 , a substituted or unsubstituted aryl group of C 6-10 , or, a substituted or unsubstituted heteroaryl group of C 5-10Heteroaryl, amido, or mercapto; f represents L 1 The linking site with L 2 h represents L 2 The linking site with P;

[0025] u is 0, 1, 2, or 3;

[0026] Each R 1-1 、R e-1 and R d-1 independently represents an alkylene group of C 1-5 .

[0027] In one embodiment, the P is a chitosan structural unit, an acrylamide structural unit, or a group represented by formula (III-1):

[0028]

[0029] (III-1),

[0030] wherein, in formula (III-1), a and c independently selected from the range of 2 - 130; b is in the range of 10 - 80, and R is selected from hydrogen and C 1-5 alkyl.

[0031] In the present invention, in the compound represented by formula (I), each alkylene group of C 1-5 is methylene, , , or .

[0032] In the present invention, each arylene group of C 6-10 is phenylene or naphthylene, such as phenylene.

[0033] In the present invention, each heteroaryl group of C 5-10 is preferably a C 5-10 heteroaryl group, wherein the heteroatoms are independently selected from one or two of N, O, and S, the number of heteroatoms is independently 1, 2, or 3, preferably the heteroatoms are N or O, the number of heteroatoms is 1 or 2, such as pyridine, pyrimidine, or pyran.

[0034] In the present invention, each alkyl group of C 1-5 is methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, or sec-butyl, such as methyl, ethyl, or isopropyl.

[0035] In the present invention, each substituted or unsubstituted arylene group of C 6-10 or, substituted or unsubstituted heteroarylene group of C 5-10 or, substituted or unsubstituted aryl group of C 6-10 or, substituted or unsubstituted C 5-10The substituents in the heteroaryl are independently C 1-3 alkyl, halogen, hydroxyl or cyano; the C 1-3 alkyl is methyl, ethyl or propyl;

[0036] The halogen is F, Cl, Br or I.

[0037] The chelating group can be a chelating group conventional in the art, preferably a chelating group formed by a bifunctional chelating agent. For example, the bifunctional chelating agent is DOTA, NOTA, NODA, NODAGA, DOTP, TETA, ATSM, PTSM, EDTA, EC, HBEDCC, DTPA, SBAD, BAPEN, DFO or TACN.

[0038] In one embodiment, the chelating group is , or .

[0039] In one embodiment, the L 1 is a linking group represented by formula (I-2) or (I-3):

[0040] (I-2),

[0041] (I-3)

[0042] wherein, in formula (I-2), R 1 is or C 1-5 alkylene, and i represents the connection to Re; R e is substituted or unsubstituted C 6-10 arylene, or, substituted or unsubstituted C 5-10 heteroarylene; wherein e represents the connection to G, and f represents the connection to L 2 .

[0043] In formula (I-3), each n is independently an integer selected from the range of 0-10, and R b is hydrogen or hydroxyl.

[0044] In one embodiment, in formula (I-2), R 1 or methylene, and i represents the connection site to R e .

[0045] In one embodiment, in formula (I-2), R e is substituted or unsubstituted C 6-10 arylene, such as phenylene;

[0046] In one embodiment, in formula (I-2), R1 is methylene or , and R e is phenyl, and i represents connection to R e .

[0047] In one embodiment, in formula (I-3), each n is independently 1, 2, 3, 4, or 5, and R b is hydroxyl.

[0048] In one embodiment, L 1 is , or , e represents connection to G, and f represents connection to L 2 .

[0049] In one embodiment, in formula (II-1), z is 2.

[0050] In one embodiment, in formula (II-2), R 2 and R 3 are both methyl.

[0051] In one embodiment, in formula (II-2), u is 0.

[0052] In one embodiment, in formula (II-2), R 2 and R 3 are both methyl, and u is 0.

[0053] In one embodiment, L 2 is (for example ), or .

[0054] In one embodiment, in formula (III-1), a and c are independently integers in the range of 80 - 130, and b is an integer in the range of 30 - 60; for example, a or c is 80 or 101, and b is 27 or 56.

[0055] In one embodiment, the acrylamide structural unit includes a group represented by formula (III-2):

[0056] (III-2)

[0057] In formula (III-2), R c is hydrogen, -CH-(CH 3 ) 2 , -C-(CH 3 ) 3 , or -(CH 2 ) m -NH 2, where m is an integer within 0 - 5, R a is independently a divalent group formed by removing one hydrogen from any of -CH-(CH 3 ) 2 , -C-(CH 3 ) 3 , or -(CH 2 ) m -NH 2 ;

[0058] p or q is independently an integer within the range of 20 - 200.

[0059] In one embodiment, in formula (III - 2), R c is -CH-(CH 3 ) 2 or -(CH 2 ) m -NH 2 , where m is 1, 2, 3, 4, or 5.

[0060] In one embodiment, in formula (III - 2), R c is -CH-(CH 3 ) 2 or -(CH 2 ) 2 -NH 2 .

[0061] In one embodiment, in formula (III - 2), R a is a divalent group formed by removing hydrogen from -(CH 2 ) m -NH 2 , where m is an integer within 0 - 5, for example -(CH 2 ) 2 -NH-.

[0062] In one embodiment, in formula (III - 2), p is an integer within the range of 100 - 150, and q is an integer within the range of 20 - 40; for example, when R c is -CH-(CH 3 ) 2 , p is 110 - 125, and q is an integer within the range of 20 - 25, or when R c is -(CH 2 ) 2 -NH 2 , p is an integer within the range of 120 - 135, and q is an integer within the range of 25 - 35.

[0063] In one embodiment, the P is , , or 。

[0064] In one embodiment, the P is derived from the polymer represented by formula (IV):

[0065]

[0066] (IV),

[0067] wherein in formula (IV), the definitions of a, b, c and R are as described above.

[0068] In one embodiment, in formula (IV), the average relative molecular weight of the polymer is 8000 - 13000 Da, such as 8600 Da - 11500 Da, and preferably 8600 Da or 11500 Da.

[0069] In some embodiments, the P is derived from the polymer represented by formula (V):

[0070] (V),

[0071] In formula (V), R c is hydrogen, -CH-(CH 3 ) 2 , -C-(CH 3 ) 3 , or -(CH 2 ) m -NH 2 , where m is an integer within the range of 0 - 5, p is an integer within the range of 100 - 150, and q is an integer within the range of 20 - 40.

[0072] In one embodiment, the polymer satisfies the following Embodiment (1) - Embodiment (5):

[0073] Embodiment (1):

[0074] The chelating group is ;

[0075] The P is ; wherein, R is methyl, a is an integer within the range of 75 - 110, and b is an integer within the range of 25 - 70;

[0076] The L 1 is , where e represents the connection to G, and f represents the connection to L 2 ;

[0077] The L 2 is , f represents the connection site of L 1 to L 2 , and h represents L2 Linking site with P;

[0078] Scheme (2):

[0079] The chelating group is ;

[0080] The P is , where R is methyl, a is an integer in the range of 75 - 110, and b is an integer in the range of 25 - 70;

[0081] The L 1 is , where e represents connection with G, and f represents connection with L 2 ;

[0082] The L 2 is , f represents the linking site of L 1 with L 2 , and h represents the linking site of L 2 with P;

[0083] Scheme (3):

[0084] The chelating group is ;

[0085] The P is , where R is methyl, a is an integer in the range of 75 - 110, and b is an integer in the range of 25 - 70;

[0086] The L 1 is , where e represents connection with G, and f represents connection with L 2 ;

[0087] The L 2 is , f represents the linking site of L 1 with L 2 , and h represents the linking site of L 2 with P;

[0088] Scheme (4):

[0089] The chelating group is ;

[0090] The P is or ;

[0091] The L 1 is , where e represents connection with G, and f represents connection with L 2 ;

[0092] The said L 2 is a chemical bond;

[0093] Scheme (5):

[0094] The said chelating group is ;

[0095] The said P is or ;

[0096] The said L 1 is a chemical bond;

[0097] The said L 2 is a chemical bond.

[0098] In some embodiments of the present invention, the chemical bond is a covalent bond.

[0099] In one embodiment, the radionuclide-labeled polymer has any of the following structures:

[0100] , ,

[0101] , or , where a, b, c, p, q, R, R a and R c are as described above. For example

[0102] , ,

[0103] , or .

[0104] In some embodiments, the radionuclide-labeled polymer has any of the following structures:

[0105] ,

[0106] ,

[0107] ,

[0108] ,

[0109] ,

[0110] ,

[0111] ,

[0112] ,

[0113] or .

[0114] The present invention also provides a polymer represented by formula A:

[0115] ;

[0116] wherein, L 1 , L 2 , and P are as defined above, Q is a chelating group, and the chelating group is as defined above.

[0117] In one embodiment, the polymer represented by formula A is any of the following structures:

[0118] , , , or

[0119] wherein a, b, c, p, q, R, Rc, and Ra are all as defined above, for example

[0120] , , , , , , , , or .

[0121] The present invention also provides a method for preparing a radionuclide-labeled polymer, which comprises the following steps: mixing and incubating the polymer represented by formula (A) as defined above with a solution containing a radionuclide to obtain a radionuclide-labeled polymer.

[0122] In one embodiment, in the preparation method, the radionuclide-labeled polymer is as defined above.

[0123] In one embodiment, the preparation method comprises the following steps:

[0124] S1. Provide an oxalic acid solution containing radioactive metal ions to obtain a mixed solution 1;

[0125] S2. Incubate the mixed solution 1 with an Na 2 CO 3 solution at room temperature to obtain a mixed solution 2;

[0126] S3. Mix the mixed solution 2 with the polymer shown in formula (A) and a buffer solution, incubate, and obtain a product after the reaction ends;

[0127] Among them, in step S1, the solvent of the oxalic acid solution is water;

[0128] In step S2, the solvent of the Na 2 CO 3 solution is water.

[0129] Preferably, the preparation method further comprises:

[0130] S0. Prepare the polymer to be labeled;

[0131] Among them, step S0 includes the following steps:

[0132] S01. Synthesize an amino derivative of a temperature-sensitive polymer;

[0133] S02. Perform a chelation reaction on the amino derivative with a chelating agent to generate the polymer shown in formula (A), where the polymer shown in formula (A) is a chelate of a polymer and a chelating group.

[0134] In one embodiment, S01 includes the following steps:

[0135] (1) In anhydrous dichloromethane, an amino-protected amino acid reacts with the above temperature-sensitive polymer in the presence of N,N'-dicyclohexylcarbodiimide and 4-dimethylaminopyridine to obtain a crude product;

[0136] (2) In anhydrous dichloromethane, perform a deprotection reaction on the crude product of step (1) to obtain an amino derivative of the temperature-sensitive polymer.

[0137] In one embodiment, S02 includes the following steps:

[0138] (3) In a bicarbonate solution, the amino derivative in step (2) undergoes an addition reaction with a chelating agent to obtain a polymer A to be labeled;

[0139] Among them, the amino-protected amino acid is valine protected by a Boc group;

[0140] The deprotection reaction is Boc deprotection;

[0141] The bifunctional chelator is one or more of DFO or DOTA.

[0142] The present invention also provides a gel composition, which comprises the radionuclide-labeled polymer and a hydrogel preparation as described above.

[0143] The hydrogel preparation in this application can be a hydrogel preparation conventionally used in the art for in vivo vascular embolization, in vivo hemostasis, drug delivery, in vivo tissue filling or in vivo tissue repair. For example, it preferably includes a temperature-sensitive gelling agent and a gelling agent.

[0144] In one embodiment, the temperature-sensitive gelling agent is one or more of poloxamer, N-isopropylacrylamide polymer, chitosan or polyethylene glycol block copolymer.

[0145] In one embodiment, the gelling agent is one or more of sodium alginate, hydroxyethyl cellulose and CaCl 2 among others.

[0146] In one embodiment, the hydrogel preparation further includes a contrast agent, such as iohexol.

[0147] In one embodiment, based on the total volume of the hydrogel preparation, the mass-volume ratio of the temperature-sensitive gelling agent is 0.12 - 0.2 g / mL, and the mass-volume ratio of the gelling agent is 0.005 - 0.5 g / mL. Preferably, the mass-volume ratio of the temperature-sensitive gelling agent is 0.14 - 0.18 g / mL, and the mass-volume ratio of the gelling agent is 0.005 - 0.3 g / mL. In one embodiment, the hydrogel preparation includes: 0.2 - 1 g / mL iohexol, 0.02 - 0.1 g / mL sodium alginate, 0.12 - 0.2 g / mL poloxamer, 0.005 - 0.1 g / mL hydroxyethyl cellulose or CaCl 2 , and water.

[0148] In one embodiment, the gelling agent is prepared by the following method: dissolving sodium alginate and hydroxyethyl cellulose in an aqueous solution of iohexol, heating to dissolve, and cooling to obtain; in the aqueous solution of iohexol, the mass ratio of iohexol to water is (1 - 5):3, such as 2:3.

[0149] In one embodiment, the hydrogel preparation includes one or more of sodium alginate, poloxamer and hydroxyethyl cellulose. Preferably, the hydrogel preparation consists of iohexol, sodium alginate, poloxamer, hydroxyethyl cellulose and water.

[0150] In one embodiment, the mass-volume ratio of the radionuclide-labeled polymer to the hydrogel preparation is 0.01 mg / mL to 2 mg / mL, preferably 0.01 mg / mL to 1.5 mg / mL, more preferably 0.06 mg / mL to 1 mg / mL, such as 0.06 mg / mL.

[0151] In one embodiment, based on the volume of the gel composition, the gel composition consists of 0.06 mg / mL of the radionuclide-labeled polymer, 0.4 g / mL of the iohexol, 0.02 g / mL of the sodium alginate, 0.14 g / mL of the poloxamer, 0.005 g / mL of the carboxymethyl cellulose, and the balance of water.

[0152] In one embodiment, the sol-gel transition temperature of the gel composition is 25 - 37 °C, preferably 25.0 - 28.3 °C, such as 25.5 - 27.5 °C.

[0153] The present invention also provides a radionuclide-labeled polymer prepared by the preparation method as described above.

[0154] The present invention also provides a tracer comprising the radionuclide-labeled polymer as described above.

[0155] In one embodiment, the tracer is prepared by adding the radionuclide-labeled polymer as described above to the hydrogel preparation as described above. The tracer is a PET / CT imaging agent or an SPECT imaging agent, and thus the in vivo distribution of the gel composition can be observed by PET / CT imaging.

[0156] In one embodiment, the tracer is used to evaluate the embolization performance of the gel composition as described above.

[0157] The present invention also provides an application of the gel composition as described above, wherein the application is the application of the gel composition in the preparation of materials for in vivo vascular embolization, in vivo hemostasis, or in vivo tissue repair.

[0158] In one embodiment, the application is the application in the preparation of an embolizing agent, and the embolizing agent can be an embolizing agent for transcatheter arterial chemoembolization for treating tumors.

[0159] The present invention also provides a method for tracing an implantable hydrogel, which comprises the following steps:

[0160] (1) Provide a hydrogel and add the radionuclide-labeled polymer as described above;

[0161] (2) Implant it into a subject;

[0162] (3) Perform PET / CT imaging at multiple preset time points to check the imaging effect and / or perform radioactive counting.

[0163] This application also provides a method for radiolabeling a hydrogel preparation, including:

[0164] Provide the polymer radiolabeled with the radionuclide as described above;

[0165] Add the polymer radiolabeled with the radionuclide to the hydrogel preparation as described above.

[0166] In one embodiment, the temperature-sensitive gelling agent and the polymer radiolabeled with the radionuclide have the same temperature-sensitive structural unit.

[0167] Preferably, when the temperature-sensitive gelling agent is poloxamer, P in the polymer radiolabeled with the radionuclide includes the structure shown in formula (III-1);

[0168] When the temperature-sensitive gelling agent is N-isopropylacrylamide polymer, P in the polymer radiolabeled with the radionuclide includes the structure shown in formula (III-2).

[0169] In one embodiment, the subject is a human, a primate or a non-primate mammal.

[0170] In one embodiment, the subject is an experimental animal model.

[0171] Except as described above, when used in the specification and claims of this application, unless otherwise specifically indicated, the following terms have the meanings shown below:

[0172] As used herein, "temperature-sensitive" refers to the property of gelation occurring from a liquid state or solidifying from a liquid to a gel or solid state with an increase in temperature. "Hydrogel preparation" refers to a water-soluble or hydrophilic polymer or a composition containing such a polymer, and also includes a gel-like substance formed by chemical cross-linking or physical cross-linking of the composition; the hydrogel preparation may include a temperature-sensitive polymer, such as poloxamer or acrylamide polymer or other known temperature-sensitive polymers in the art, so the hydrogel preparation has temperature-responsive properties, such as transforming from a sol state to a gel state at a certain temperature; the hydrogel preparation can be used as a material for in vivo vascular embolization, in vivo hemostasis, drug delivery, in vivo tissue filling or in vivo tissue repair. The hydrogel preparation can be any hydrogel material for medical, pharmaceutical or cosmetic use.

[0173] As used herein, "phase transition temperature" refers to the temperature or temperature range at which a polymer undergoes a phase transition.

[0174] The term "polymer" refers to a molecule formed by the combination of two or more oligomer units. Chemical units are usually connected by covalent bonds. Two or more combined units in a polymer can be all the same, and in this case, the polymer is called a homopolymer. They can also be a combination of different units, and these polymers are called copolymers.

[0175] The term "chemical bond" refers to the general term for the strong interaction forces between two or more adjacent atoms (or ions) within a pure substance molecule or crystal, generally including ionic bonds, covalent bonds, and metallic bonds.

[0176] The term "linking group" refers to the remaining part after an organic compound that plays a linking role loses one or more atoms, and it is the general term for atomic groups and radicals that play a linking role. The linking group contains usually covalent bonds inside, but the linking group is not a chemical bond.

[0177] The term "heteroaryl" refers to an aromatic system containing heteroatoms, and the number of said heteroatoms is 1, 2, 3, or 4, and the types of heteroatoms are selected from N, O, or S. Among them, heteroaryl groups include but are not limited to: thiazolyl, thiophenyl, pyridyl, pyrimidinyl.

[0178] On the basis of not violating the common knowledge in this field, the above preferred conditions can be combined arbitrarily to obtain various preferred examples of the present invention.

[0179] The reagents and raw materials used in the present invention are all commercially available.

[0180] The beneficial effects of the present invention are as follows: The present invention provides a radionuclide-labeled polymer, which can be used to mix with a thermosensitive hydrogel without changing the physical and chemical properties of the hydrogel itself, and can still maintain the original viscosity and thermosensitive curing performance of the hydrogel. The radionuclide-labeled signal can exist stably in the body for a long time, maintain a long half-life, and the continuous imaging period can reach more than 11 days. It can be used for in vivo tracing of embolizing agents in the hydrogel, and the distribution status of the hydrogel embolization in the body can be conveniently and continuously monitored without multiple angiographies.

[0181] The radionuclide-labeled polymer of the present invention is covalently connected to the polymer unit through a chelating group, and then chelates the radionuclide through the chelating group, and has a better binding force with the hydrogel preparation. When used for radioactive labeling of implantable hydrogels, compared with the method of directly adding zirconium salt chelation or non-covalent connection such as hydrogen bonds, it has better stability and in vivo imaging time. BRIEF DESCRIPTION OF THE DRAWINGS

[0182] Figure 1 For dog B01 89 Dynamic transverse sectional view of Zr-labeled poloxamer in vivo for 5 h;

[0183] Figure 2For dog B01 89 Dynamic coronal plane images of Zr-labeled poloxamer at 5 h in vivo;

[0184] Figure 3 For dog B01 89 Dynamic transverse plane images of Zr-labeled poloxamer at 1 d in vivo;

[0185] Figure 4 For dog B01 89 Dynamic coronal plane images of Zr-labeled poloxamer at 1 d in vivo;

[0186] Figure 5 For dog B01 89 Dynamic transverse plane images of Zr-labeled poloxamer at 5 d in vivo;

[0187] Figure 6 For dog B01 89 Dynamic coronal plane images of Zr-labeled poloxamer at 5 d in vivo;

[0188] Figure 7 For dog B01 89 Dynamic transverse plane images of Zr-labeled poloxamer at 7 d in vivo;

[0189] Figure 8 For dog B01 89 Dynamic coronal plane images of Zr-labeled poloxamer at 7 d in vivo;

[0190] Figure 9 For dog B01 89 Dynamic transverse plane images of Zr-labeled poloxamer at 11 d in vivo;

[0191] Figure 10 For dog B01 89 Dynamic coronal plane images of Zr-labeled poloxamer at 11 d in vivo;

[0192] Figure 11 For dog B04 89 Dynamic transverse plane images of Zr-labeled poloxamer at 5 h in vivo;

[0193] Figure 12 For dog B04 89 Dynamic coronal plane images of Zr-labeled poloxamer at 5 h in vivo;

[0194] Figure 13 For dog B04 89 Dynamic transverse plane images of Zr-labeled poloxamer at 1 d in vivo;

[0195] Figure 14 For dog B04 89 Dynamic coronal plane images of Zr-labeled poloxamer at 1 d in vivo;

[0196] Figure 15 For dog B04 89 Dynamic transverse plane images of Zr-labeled poloxamer in dog B04 in vivo at 5 days;

[0197] Figure 16 For dog B04 89 Dynamic coronal plane images of Zr-labeled poloxamer in dog B04 in vivo at 5 days;

[0198] Figure 17 For dog B04 89 Dynamic transverse plane images of Zr-labeled poloxamer in dog B04 in vivo at 7 days;

[0199] Figure 18 For dog B04 89 Dynamic coronal plane images of Zr-labeled poloxamer in dog B04 in vivo at 7 days;

[0200] Figure 19 For dog B04 89 Dynamic transverse plane images of Zr-labeled poloxamer in dog B04 in vivo at 11 days;

[0201] Figure 20 For dog B04 89 Dynamic coronal plane images of Zr-labeled poloxamer in dog B04 in vivo at 11 days;

[0202] Figure 21 For 89 Calibration curve of the detection values by Zr activity meter and γ counter. Specific implementation mode

[0203] The present invention will be further described below by way of examples, but the present invention is not limited to the scope of the described examples. The experimental methods without specific conditions in the following examples are carried out according to conventional methods and conditions, or selected according to the product specifications.

[0204] Example:

[0205] Materials:

[0206] Polymer 1:

[0207]

[0208] Manufacturer: BASF CORPORATION, model Kolliphor® P 407Geismar.

[0209] Polymer 2:

[0210]

[0211] Manufacturer: BASF CORPORATION, model Kolliphor® P 188 Geismar.

[0212] Chitosan: average relative molecular weight 5000 Da, product number C434552, non-animal origin, purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.

[0213] Iohexol: Manufacturer: Shanghai Stell Pharmaceutical Co., Ltd.

[0214] Sodium alginate: Manufacturer: Qingdao Mingyue Alginate Tissue Engineering Materials Co., Ltd.

[0215] 89 Zr:

[0216] Manufacturer: PerkinElmer, Inc.

[0217] Purity and content: After opening the package, the total activity concentration was detected to be 19.54 mCi / mL.

[0218] Prescription composition: 89-Zr is dissolved in 1 mol / L oxalic acid solution.

[0219] Storage conditions: At room temperature, stored in a lead container in a 2 mL V-bottom screw-cap glass bottle, pay attention to preventing ionizing radiation.

[0220] N-isopropylacrylamide monomer: sourced from Shanghai Aladdin, product number I106818.

[0221] N-(3-aminopropyl) methacrylamide monomer (APAM monomer): Shanghai Aladdin, product number N129096.

[0222] Animal source of Beagle dogs: Nanjing Chaimen Biotechnology Co., Ltd.

[0223] Other reagents are commercially available unless otherwise specified.

[0224] Preparation Example 1 Radioactive Labeled Polymer 89 Preparation of Zr-DFO-P1-1

[0225] 1.1 Synthesis of Thermosensitive Polymer and DFO Chelate

[0226] (1) Polymer Amino Activation

[0227] 1.086 g of Boc-Val-OH (Boc-L-valine), 1.91 g of DCC (N,N'-dicyclohexylcarbodiimide), and 1.22 g of DMAP (4-dimethylaminopyridine) were dissolved in 40 mL of ultradry anhydrous DCM (dichloromethane) in a molar ratio of 1:2:2. Under argon protection, the reaction was carried out at room temperature for half an hour. Then, 5.75 g of polymer 1 (P1-1) was added, and the reaction continued for 2 days. The DCM was concentrated by rotary evaporation, precipitated with ice-cold diethyl ether, and the white precipitate was collected by filtration. The precipitate was redissolved in 3 mL of DCM and then precipitated again with ice-cold diethyl ether. The precipitation and filtration were repeated three times. The precipitate was redissolved in ultrapure water at a ratio of 1 mg:1 mL of precipitate to ultrapure water (heating), dialyzed for three days, and freeze-dried to obtain 4.13 g of Boc-group protected polymer P1-1-Val-Boc with a yield of 71.9%.

[0228] (2) Deprotection of Boc

[0229] 4 g of polymer P1-1-Val-Boc was dissolved in 20 mL of DCM and placed in an ice-water bath. Under argon protection, 45 mL of trifluoroacetic acid was added, and the reaction was stirred for 2 hours. The solvent was removed by rotary evaporation, the product was redissolved in chloroform, and the product was dried by rotary evaporation and dissolved in hot water for dialysis for three days. After freeze-drying, 3.2 g of the deprotected polymer P1-1-Val was obtained with a yield of 80%.

[0230]

[0231] (3) Preparation of DFO chelate

[0232] 15.3 mg of polymer P1-1-Val was dissolved in 4 mL of 0.1 M NaHCO 3 (sodium bicarbonate) solution, and p-SCN-Bn-DFO (CAS No.: 1222468-90-7) (2 mg; 200 μL DMSO) was added. The molar reaction ratio of polymer P1-1-Val to p-SCN-Bn-DFO was 1:2 and was slowly added dropwise. The reaction was carried out at room temperature for 1 h, dried by rotary evaporation, dissolved in hot water for dialysis for three days, and after freeze-drying, 10.2 mg of polymer DFO-P1-1 was obtained with a purity of 99.3% and a yield of 66.7%.

[0233] .

[0234] 1.2 89 Synthesis of Zr-labeled polymer

[0235]

[0236] 89 Zr-DFO-P1-1

[0237] (1) Preparation of labeling solution

[0238] 89 Oxalic acid solution of Zr: Zirconium 89 Zr] is dispersed in 1M oxalic acid solution (pH < 4) (PerkinElmer, Inc);

[0239] Preparation of 0.5 M HEPES (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid) buffer solution: First, add 18 mL of ultrapure water to 20 mL of 1M HEPES buffer solution and measure its pH value. If the pH value < 7.1, use 1M NaOH to adjust its pH value; if pH > 7.3, use 1M H 2 SO 4 to adjust its pH value. When the pH value of the HEPES buffer solution is within the range of 7.1 - 7.3, continue to add a certain volume of ultrapure water to make the total volume 40 mL.

[0240] Preparation of polymer DFO-P1-1 solution (1 mg / mL): Accurately weigh 0.6 mg of polymer DFO-P1-1 and dissolve it in 600 µL of 10 mM pH 8.4 borate buffer solution.

[0241] (2) 89 Zr labeling reaction

[0242] A. Accurately measure 84 μL of 89 Zr oxalic acid solution into a glass reaction flask;

[0243] B. Add 1M oxalic acid solution to the glass reaction flask to make the total volume in the reaction flask 200 μL;

[0244] C. Use a pipette to accurately aspirate 90 μL of 2M Na 2 CO 3 solution into the reaction flask and incubate at room temperature for 3 min;

[0245] D. While gently shaking the reaction flask, dissolve 0.3 mL of 0.5M HEPES (pH 7.1 - 7.3) and 600 µL of the prepared polymer DFO-P1-1 solution (1 mg / mL), and then add 0.7 mL of 0.5M HEPES to the reaction flask in sequence (Note: The pH value of the reaction solution should be between 6.8 - 7.2 to obtain the best labeling efficiency, and at the same time, metal needles of syringes are prohibited for solution preparation);

[0246] E. Incubate at room temperature for 1 h while gently shaking the reaction flask;

[0247] F. After the reaction is completed, use an ultrafiltration centrifuge tube to purify the reaction product to remove free89 Zr (Washing solution: 10 mM pH 8.4 borate buffer solution; 500 µL * 2 times).

[0248] Preparation Example 2 Radioactively Labeled Polymer 89 Preparation of Zr-DFO-P1-2

[0249]

[0250] Only replace Polymer 1 (P1-1) in Preparation Example 1 with Polymer 2 (P1-2), calculate its dosage according to the average relative molecular mass of Polymer 2, and carry out the reaction according to the steps of Preparation Example 1.

[0251] Preparation Example 3 Radioactively Labeled Polymer 89 Preparation of Zr-DOTA-P1-1

[0252]

[0253] Only replace p-SCN-Bn-DFO (CAS No.: 1222468-90-7) in Preparation Example 1 with p-SCN-Bn-DOTA (CAS No.: 127985-74-4), and carry out the reaction according to the steps of Preparation Example 1.

[0254] Preparation Example 4 Radioactively Labeled Polymer 89 Preparation of Zr-DFO-P2

[0255] Dissolve chitosan in 0.1 M acetic acid solution, add p-SCN-Bn-DFO (2 mg; 200 μL DMSO), slowly dropwise add the reaction of chitosan and p-SCN-Bn-DFO at a molar ratio of 1:2, react at 37 °C for 4 h, spin dry and dissolve in hot water for dialysis for three days, and obtain the product DFO-P2 after freeze-drying. Then carry out according to the steps of Step 1.2 in Preparation Example 1 89 Radioactive labeling of Zr 89 Zr-DFO-P2

[0256] Preparation Example 5 Radioactively Labeled Polymer 89 Preparation of Zr-DOTA-P3-1

[0257] (1) Synthesis of Boc-protected N-isopropylacrylamide (NIPAM) polymer:

[0258] 3.69 g of N-isopropylacrylamide monomer (NIPAM), 1.76 g of N-hydroxysuccinimide (NHS) and 290 mg of azobisisobutyronitrile (AIBN) were dissolved in 10 mL of THF. The mixture was reacted at 60 °C for 12 h to prepare pNIPAM-NHS. Then 80.6 g of N-Boc-ethylenediamine was added and reacted for 12 h. Then it was precipitated with 150 mL of ether, washed twice with ether after filtration, and then dried to obtain pNIPAM-Boc with a yield of 83%;

[0259]

[0260] (2)Deprotection of Boc group:

[0261] 2.4 g of pNIPAM-Boc was dissolved in 50 mL of aqueous trifluoroacetic acid solution (volume ratio of trifluoroacetic acid to water was 8:2), stirred at room temperature for 2 h, dried by vacuum rotary evaporation, redissolved in water, purified by Sephadex G25 chromatography column, and freeze-dried to obtain pNIPAM with a yield of 90%;

[0262]

[0263] (3)Activation of DOTA: 48 mg of DOTA was dissolved in 2 mL of water, 9.0 mg of 1-ethyl-(3-dimethylaminopropyl)carbodiimide (EDC) was dissolved in 0.5 mL of water. The two aqueous solutions were mixed to form a reaction mixture, and the pH of the reaction mixture was adjusted to 5.0 with 0.1 M NaOH. Then 6.8 mg of NHS was added, and the mixture was stirred in an ice bath for 40 minutes to obtain DOTA-NHS;

[0264]

[0265] (4)Synthesis of pNIPAM-DOTA compound: 298 mg of pNIPAM (containing 0.03 mmol of amino groups) was dissolved in a mixed solvent, which included 3 mmol of N,N-diisopropylethylamine (DIEA) and 6 mL of DMAC. It was heated to 45 °C, 0.03 mmol of DOTA-NHS was added, and the reaction was carried out for 3 h. Purification was carried out by centrifugal filtration method, and the cut-off molecular weight of the centrifugal filtration membrane was 3.5 kDa. After purification, it was freeze-dried to obtain the chelating ligand product DOTA-P3-1;

[0266] 。

[0267] (5)Zr radiolabeling was carried out according to Step 1.2 of Preparation Example 1 to obtain 89 Zr-DOTA-P3-1. 89 Zr-DOTA-P3-1.

[0268] 。

[0269] In this preparation example, the N-isopropylacrylamide polymer (the product of step (2) of Preparation Example 5): The average molecular weight Mn measured by size exclusion chromatography (SEC) was 17,400 g / mol.

[0270] After connecting DOTA, in D 2 O solution, 1H-NMR in D 2 O solution was recorded using a Varian AC400 NMR spectrometer (Varian NMR Systems, Palo Alto, CA). From the proton spectrum, the molar ratio p:q of the two repeating units was calculated to be about 0.85:0.15. 1 H-NMR, and from the proton spectrum, the molar ratio p:q of the two repeating units was calculated to be about 0.85:0.15.

[0271] Preparation Example 6 Radiolabeled Polymer 89 Preparation of 89Zr-DFO-P3-2

[0272] (1) 3.37 g of N-(3-aminopropyl)methacrylamide monomer (APAM monomer) was added, and 164 mg of azobisisobutyronitrile (AIBN) was added as an initiator. Free radical reaction was carried out in an acetone-DMSO mixed solvent (V(acetone):V(DMSO)=15:1). Polymerization reaction was carried out at 55 °C for 24 hours under vacuum conditions. The precipitate was recovered by filtration to obtain the crude product. The crude product was dissolved in a small amount of methanol, and 15 times the volume of excess acetone was added for precipitation. After vacuum drying, polyaminopropylmethacrylamide was obtained. The average molecular weight Mn measured by size exclusion chromatography (SEC) was 19,200 - 21,500 g / mol, using a Synchropak CATSEC-300 chromatographic column.

[0273] (2) 2 mg (200 μL DMSO) of p-SCN-Bn-DFO (CAS No.: 1222468-90-7) was slowly added dropwise to 12 mg of polyaminopropylmethacrylamide. Sodium hydroxide was added to adjust the pH to 6.0, and the reaction was carried out at room temperature for 1 h. After rotary evaporation, it was dissolved in water and dialyzed for three days. The cut-off molecular weight of the centrifugal filter membrane was 10 kDa. After freeze-drying, 9.7 mg of the polymer was obtained, and the yield was 69.3%. 1H-NMR in D 2 O solution was recorded using a Varian AC400 NMR spectrometer (Varian NMR Systems, Palo Alto, CA). From the proton spectrum, the molar ratio p:q of the two repeating units was calculated to be about 0.82:0.18. 2 O solution was recorded using a Varian AC400 NMR spectrometer (Varian NMR Systems, Palo Alto, CA). From the proton spectrum, the molar ratio p:q of the two repeating units was calculated to be about 0.82:0.18. 1 H-NMR, and from the proton spectrum, the molar ratio p:q of the two repeating units was calculated to be about 0.82:0.18.

[0274] (3) Follow the procedure of Step 1.2 in Preparation Example 1 89 for Zr radiolabeling to obtain 89 Zr-DFO-P3-2.

[0275] .

[0276] Preparation Example 7 Radiolabeled Polymer 89 Preparation of Zr-DFO-LP1-1

[0277] Replace p-SCN-Bn-DFO in Preparation Example 1 with Compound D-1 (CAS: 1623757-39-0), react P1-Val with D-1 to prepare a DFO-conjugated polymer, and then follow the procedure of Step 1.2 in Preparation Example 1 89 for Zr labeling.

[0278]

[0279] Preparation Example 8: Preparation of 89 Zr-labeled Hydrogel Preparation

[0280] (1) Preparation 1

[0281] Prepare an iohexol aqueous solution: Accurately measure iohexol and water for injection according to a mass ratio of 2 g:3 g, mix evenly to prepare a 40% iohexol solution.

[0282] Prepare a solution of sodium alginate and hydroxypropyl methylcellulose: Weigh 2 g of sodium alginate and 0.5 g of hydroxypropyl methylcellulose (HPMC-100M), slowly add sodium alginate and hydroxypropyl methylcellulose to 100 mL of 40% iohexol solution, slowly heat to 35 °C at room temperature to promote the dissolution of sodium alginate and hydroxypropyl methylcellulose, and let it cool to room temperature after complete dissolution.

[0283] Prepare a gel composition: Accurately weigh 14 g of poloxamer 407, slowly add it to 100 mL of the prepared sodium alginate and hydroxypropyl methylcellulose solution, let it stand at 4 °C for 3 - 4 h until completely dissolved to obtain Preparation 1. The sol-gel phase transition temperature is 26.5 °C, and it gels within 32 seconds and transforms into a solid within 50 seconds at 37 °C; the viscosity is 0.9 Pa·S (25 °C), 1.2×10 3 Pa·S (30 °C).

[0284] (2) Preparation 2: Embrace HES polyethylene glycol liquid embolization agent.

[0285] (3) Preparation 3: Taipu Shen TM Temperature-sensitive liquid embolization agent (copolymer of N-isopropylacrylamide and N-n-propylacrylamide).

[0286] (4) Preparation of zirconium-labeled gel composition:

[0287] Preparation 89 Zr polymer-labeled preparation: Add 1 mL of the 89 Zr-labeled polymer solution (containing 0.6 mg of polymer) prepared in Preparation Examples 1-7 to 10 mL of Preparation 1, Preparation 2, or Preparation 3, mix slightly, and refrigerate at 4-8 °C after stirring.

[0288] (5) Preparation 89 Zr chelator-free labeled preparation: Add 1 mL of the oxalic acid solution of the radioactive isotope 89 Zr to 10 mL of Preparation 1, Preparation 2, or Preparation 3 and mix.

[0289] Test Example 1: Performance test

[0290] 1.1 89 Radiochemical purity of Zr-labeled hydrogel preparation

[0291] Using a Mini-Scan thin-layer scanner (Eckert & Ziegler Radiopharma Inc, USA), with a fast thin-layer chromatography paper (Agilent ITLC strip 1 cm × 10 cm, USA) as the carrier, after spotting, develop with a 20 mM citric acid solution (pH 4.9-5.1) developing system, dry the chromatography paper, and then collect data with a thin-layer scanner. After data collection is completed, obtain an image for data analysis.

[0292] The performance test results of the gel compositions prepared from the polymers and hydrogel preparations prepared in the preparation examples according to the ratio in Preparation Example 8.(1) are shown in Table 1 below:

[0293] Table 1

[0294]

[0295] In this experiment, the radiochemical purity of the 89 Zr-labeled hydrogel preparation and the labeling stability at different time points were evaluated by thin-layer chromatography. As shown in the above table, 89 the radiochemical purity of the Zr-labeled hydrogel preparation remained above 93% at different time stages, indicating 89 that the Zr-labeled hydrogel preparation had good in vitro stability. In particular, after the Zr-labeled polymer was used for labeling the hydrogel preparation, the radiochemical purity was greater than 93% and met the experimental requirements, and the radiochemical purity was still greater than 93% after 19 days of storage. In the experimental group numbered h, the oxalic acid solution of 89 Zr was directly added, 89 and 89The Zr ions chelate with the hydroxyl groups of sodium alginate and promote the crosslinking of sodium alginate to form a network structure, which 89 coats Zr, achieving 89 chelator-free coating of Zr. At 1 day after initial labeling, there is a high labeling efficiency, but as the storage time extends, the radiochemical purity decreases, indicating that 89 the content of Zr-labeled sodium alginate becomes less, and more 89 Zr detaches from the labeled substance, and the stability of the experimental group numbered h is poor. The polymer provided in this experimental example has better stability due to the covalent chemical chelation of the chelating group with the polymer, and is not easily degraded after complexing radioactive metal elements through the chelating group, reducing the release of free radioactive elements and extending the radioactive labeling and imaging time.

[0296] 1.2 89 Rheological properties of the sol-gelation of the Zr-labeled gel composition

[0297] The tests were carried out on an AR-1000 constant stress rheometer (manufacturer: TA Instruments) with built-in temperature and gap calibration.

[0298] The rheometer is equipped with a cone / plate geometry (diameter 40 mm, cone angle 4°). The degassed sample is dispensed onto a temperature-controlled peltier plate and pre-equilibrated to 25 °C. During the rheological test, the sample is covered with a water-containing solvent trap to prevent water loss from the sample. During the test, the temperature of the peltier plate is controlled within ±0.1 °C of the preset temperature. Before the test, the geometric gap is calibrated. After loading an excess of the sample, the geometry is lowered to this pre-determined gap and the excess sample is removed. The sample is allowed to equilibrate for 30 s before starting the test. TA data analysis software is used to process the data.

[0299] Viscosity tests of various hydrogel formulations were evaluated using flow rheology. To determine how the hydrogels behave under increasing shear stress, steady-state flow experiments were carried out at 25 °C. The viscosity of the samples was measured in the shear stress range of 1 Pa to 100 Pa. The thermoresponsiveness of various hydrogel formulations was rheologically evaluated by oscillatory measurements. A temperature scan from 10 °C to 40 °C was carried out on all hydrogel formulations. The sol-gel transition temperature is defined as the temperature at which gelation occurs. The gel point is defined as the temperature at which the storage modulus (G’) is equal to the loss modulus (G”).

[0300] Therefore, gelation is considered to have occurred when G’ > G”. The temperature is increased at a rate of 1 °C / minute while the oscillatory pressure and angular frequency remain constant.

[0301] The performance detection results of the gel composition prepared from the polymer and hydrogel formulations prepared in the preparation examples are shown in Table 2 below.

[0302] Table 2

[0303]

[0304] It can be seen that the addition of the radioactively labeled polymer has no obvious effect on the viscosity and phase transition temperature of the liquid embolization agent, and does not affect the thermosensitive property of the rheological property of the hydrogel preparation itself. Especially when the radioactively labeled polymer and the thermosensitive material of the hydrogel preparation have the same or similar structural units, the radioactively labeled polymer and the hydrogel preparation have excellent compatibility and can be more uniformly and stably distributed in the hydrogel preparation, with less influence on the viscosity and phase transition temperature of the hydrogel preparation itself.

[0305] Test Example 2 In Vivo Test on Animals

[0306] In this test, the content of the radioactively labeled polymer in the animal blood was observed, and signals within the window width range of 110 keV ± 15% were collected by PET / CT tomography to dynamically observe the 89 dynamic distribution of the

[0307] The Beagle dogs were under general anesthesia (1.5% sodium pentobarbital, 20 mg / kg). The femoral artery was bluntly dissected, and the femoral artery was punctured under direct vision. A 5F arterial sheath was inserted, and the femoral artery and the arterial sheath were fixed with medical absorbable sutures. Under the guidance of a digital subtraction angiography machine (DSA), a 4F angiography catheter was inserted into the femoral artery to the abdominal aorta. A Y-valve was connected to the end of the 4F angiography catheter, and a 2.4F microcatheter was selectively inserted into the common hepatic artery from the positive port of the Y-valve. The gel was injected through the microcatheter, and gel reflux was avoided as much as possible. After embolization, the catheter and the arterial sheath were removed, and the upper part of the femoral artery puncture site was ligated, and the muscle and skin were sutured layer by layer.

[0308] The hydrogel preparation prepared in the preparation example was used as an in vivo vascular embolization agent and implanted into Beagle dogs. The implantation protocol is shown in Table 3:

[0309] Table 3 Implantation Protocol

[0310]

[0311] 2.1 In Blood 89 Zr Detection

[0312] The dosage of the embolizing agent was 1 mL of the labeled gel per animal, the specific activity of the gel was 280 μCi / mL, and the dosage per dog was approximately 280 μCi / dog. At 5 h, 1 d, 3 d, 5 d, 7 d, and 11 d after hepatic artery embolization, blood was collected from the cephalic vein of the forelimb of the animal using a heparin sodium anticoagulant negative pressure blood collection tube, and the blood collection volume was approximately 2 mL. After blood collection, the blood samples were detected and analyzed using a WIZARD γ counter. Establish 89 a calibration curve for the detection values (μCi) of the 89 Zr activity meter and the detection values (CPM) of the γ counter for the numerical conversion between activity and CPM. The calibration curve is shown in Figure 21 .

[0313] Based on the systemic circulation blood volume and blood leakage count results of the dogs, as well as the animal administration dosage and body weight data, estimate the 89 Zr dosage in the blood, and calculate the percentage of the radioactive activity of 89 Zr in the blood accounting for the total dosage. The circulating blood volume of Beagle dogs is 85 mL / kg. Calculate the systemic blood volume based on the animal body weight, then calculate the total blood exposure amount based on the systemic blood volume, and calculate the blood exposure percentage by comparing with the total drug amount after attenuation. The percentage of the blood exposure dosage accounting for the total dosage = body weight kg * 85 mL / kg * blood detection value (μCi / g) / administration dosage, and the blood detection value is corrected to the time of administration.

[0314] Table 4 89 Zr radioactive activity in animal blood 89

[0315]

[0316] Table 5 89 Zr radioactive percentage in animal blood 89

[0317]

[0318] As can be seen from Table 4 and Table 5, on the first day of embolization, the percentage of the radioactive activity of 89 Zr in the blood of dogs B01 - B14 was less than 7%, and then gradually decreased. By 7 days after administration, the percentage of the radioactive activity of 89 Zr in the blood of dogs B01 - B13 accounted for about 0.69% - 2% of the total administration dosage.

[0319] Generally, it shows that the proportion of the radioactive activity of 89 Zr in the blood is relatively low, indicating that the gel formed by the test substance mainly remains in the embolized site in the body, and at the same time, it also proves that the test substance has good stability after labeling and long-term placement. Specifically, after implanting a hydrogel preparation containing a 89 Zr-labeled polymer in the animal body, no free 89 Zr was found in its blood.​​​​​​​​​​​​​An abnormal increase or decrease in Zr indicates better stability in the animal body and no obvious free state.

[0320] The polymer provided in this test example 89 Zr is chemically chelated with the polymer through a chelating group, which is more stable than the preparation labeled by 89 Zr oxalate blending, is not easily degraded, and reduces the release of free radioactive elements.

[0321] In the gel implanted in Dog No. B14 89 Zr is directly mixed and coated without chemically chelating with the thermosensitive polymer. The radioactivity in the blood after implantation is significantly higher than that of other implants including 89 the gel of Zr chemically chelated polymer within 5 h - 11 d, and the radioactive signal in the blood is relatively high within 1 - 5 d, indicating that radioactive elements continuously enter the blood circulation. After 5 d, most of the 89 Zr has entered the blood and there is no more 89 Zr escaping. Therefore, the radioactivity in the blood decreases after 7 d. The radioactivity entering the blood circulation within 11 d accounts for 60% of the total dose, indicating that directly adding free 89 Zr results in poor stability of the radioactive label, easy dissociation from the embolization position, and the radioactive label cannot stably trace the position of the hydrogel for a long time.

[0322] In animals implanted with 89 the hydrogel preparation of Zr-labeled polymer, different structures of polymers as radioactive labels of the hydrogel preparation all show relatively low levels of blood radioactivity, indicating good stability. Especially when the hydrogel preparation and the radioactive-labeled polymer contain the same thermosensitive polymer units, the hydrogel preparation and the radioactive-labeled polymer have more similar viscosities and phase transition temperatures, and they can be better compatible and closely combined, and the radioactive label shows excellent stability. For example, when the radioactive-labeled polymer containing an alkyl ether structural unit 89 Zr-DFO -P1-1 or 89 Zr-DFO -P1-2 is added to Preparation 1 or 2, the total proportion of radioactive counts in the blood during the detection period is significantly lower than that added to Preparation 3; the radioactive-labeled polymer 89 Zr-DFO -P3-2 shows better stability when added to Preparation 3 with an acrylamide polymer as the thermosensitive component than when added to Preparation 1.

[0323] Comparing Animal No. B01 with Animal No. B11, only the linking group of the radioactive polymer in the preparation is different, but the tracer effect of Animal No. B01 is better, indicating that 89 Zr-DFO -P1-1 is better than 89Zr-DFO-LP1-1 has better effects, which may be due to 89 the chain length of the linking group in Zr-DFO-LP1-1.

[0324] 2.2 In Vivo Dynamic Imaging Study

[0325] Collection time: 5 h, 1 d, 5 d, 7 d, 11 d, and 18 d after embolization.

[0326] Collection method: Nuclear medicine tomography + localization CT. After the animals were anesthetized and bound, they were placed under the Discovery VCT instrument. First, a whole-body CT scan was performed to assist in the confirmation of tissues and organs, and then a whole-body nuclear medicine signal scan from the head to the bottom of the lower limbs was performed.

[0327] PET-CT imaging data processing: The coronal and cross-sectional images of the PET-CT imaging results of dogs numbered B01 - B14 were saved. Based on the imaging images and nuclear medicine images in both positions, regions of interest (ROIs) were outlined (the main organs with nuclear medicine signals were outlined), and the corresponding regions were circled, and the percentage of the count in the outlined region in the whole-body count was recorded.

[0328] The results are as Figures 1 - 20 shown in Table 6.

[0329] Figure 1 、 Figure 3 、 Figure 5 、 Figure 7 、 Figure 9 are the cross-sectional images of dog B01 at 0.5 h, 1 d, 5 d, 7 d, and 11 d after implanting the radioactively labeled hydrogel preparation, respectively; Figure 11 、 Figure 13 、 Figure 15 、 Figure 17 、 Figure 19 are the cross-sectional images of dog B04 at 0.5 h, 1 d, 5 d, 7 d, and 11 d after implanting the radioactively labeled hydrogel preparation, respectively. In the cross-sectional images, the CT localization map is in the upper left corner of each figure, the nuclear medicine imaging map of the localization position is in the upper right corner, the image of the enriched region of the cross-sectional nuclear medicine image is in the lower left corner, and the whole-body nuclear medicine signal image (this image does not change with the change of the CT localization map) is in the lower right corner. The localization of each region is at the same position - the liver.

[0330] Figure 2 、 Figure 4 、 Figure 6 、 Figure 8 、 Figure 10 are the coronal plane images of dog B01 at 0.5 h, 1 d, 5 d, 7 d, and 11 d after implanting the radioactively labeled hydrogel preparation, respectively.Figure 12 , Figure 14 , Figure 16 , Figure 18 , Figure 20 are the transverse images of Dog No. B04 at 0.5 h, 1 d, 5 d, 7 d, and 11 d after implanting the radiolabeled hydrogel preparation, respectively. The coronal plane images, with the CT localization map in the upper left corner of the image, the nuclear medicine imaging map of the localization position in the second from the left, the image of the enriched area of the coronal nuclear medicine image in the third from the left, and the whole-body nuclear medicine image in the fourth from the left. The localization of each area is at the same position - the liver.

[0331] It can be seen from the images that for the animal (B01) implanted with Scheme A, its radioactive signal was mainly distributed in the liver at 5 h in the initial stage of implantation, with a small amount in the intestine and bladder. During the period from 1 to 11 d after implantation, the radioactive signal was still mainly distributed in the liver, and only a very small amount was distributed in other organs. Compared with B01, for the animal (B04) implanted with Scheme D, its radioactive signal was mainly distributed in the liver, but the signal distribution in the lungs, intestines, kidneys, and brain was significantly more than that in B01.

[0332] Table 6 below records the average percentage of the total body count in the five regions with higher counts in the liver, lungs, intestines, kidneys, and brain of 5 dogs.

[0333] Table 6 Percentage of Organ Counts of Each Animal in the Total Body Count (%)

[0334]

[0335] It can be seen from Table 6 that on the day of administration, the proportion of the drug in the liver of each animal reached more than 80%, indicating that the test substance was embolized in the liver on the day of administration, and most of the radioactive signals were at the embolized site. The labeling efficiency of the test substance was relatively high, and there was almost no degradation. At 1 - 5 d after administration, the proportion of the liver in the liver decreased, probably because part of the test substance was embolized near the porta hepatis, and the radiolabeled polymer that was not completely combined with the hydrogel during the administration stage was washed away by the blood flow and migrated to other tissues such as the intestine, lungs, and kidneys for enrichment during this period. Therefore, slight distribution was also visible in the intestine, lungs, and kidneys at 5 - 7 d, and the proportion of radioactive counts showed a slightly increasing trend; however, the proportion of liver counts still accounted for more than 80%, still being able to significantly label the administration embolization position without affecting the evaluation of the embolization state. From the 11th to the 18th day after intervention, the proportion of the test substance in the liver gradually increased to 100% from the 5th to the 18th day after administration, and the signals in the intestinal wall and the remaining tissues and organs decreased significantly. By the 18th day, the radioactive elements in other organs had been metabolized out of the body and decayed to no signal, and only the liver showed a signal, indicating that the test substance remained at the embolized original site in the body, and 89 the Zr-labeled polymer had good long-term stability in the body.

[0336] During the entire experimental period, 89The Zr-labeled polymer is mainly enriched in the liver, and the average proportion ranges from 80% to 100%, indicating that the vast majority of radionuclides are tightly bound to the implant and do not detach, and can be significantly distinguished from the weak free signal. It is an excellent tracer for implantable hydrogels.

[0337] Specifically, for animal B01 implanted with addition plan A, the radioactive signal in the liver was significantly higher starting from 5 h, and showed a high radioactive intensity in the liver throughout the monitoring period of 1 - 18 d, all above 80% or 84%. The radioactive signals in other organs such as the lungs, intestines, kidneys, and brain were very low, indicating that in animal B01, the radiolabeled polymer was tightly bound to formulation 1 and embolized in the liver. The embolization formulation had good stability, and at the same time, the radiolabeled polymer also maintained good stability and rarely detached from the embolization site. For animal B02 implanted with addition plan B and animal B05 implanted with addition plan E, the radioactive count in the liver could also remain above 80% throughout the monitoring period. In contrast, for animal B04 implanted with addition plan D, the concentration of the radioactive signal in the liver was relatively low, starting at 80.9% in the initial stage after implantation, indicating that after addition plan D was implanted into the body, a small amount of radioactive signal was free to enter the blood and circulate to other organs. However, by the 18th day, the radioactivity in other organs outside the liver of animal B04 had also been metabolized out of the body. It can be seen that the polymer 89 Zr-DFO-P1-1 has stronger stability compared to the polymer directly formed by the direct reaction of the amino group of chitosan 89 Zr-DFO-P2 structure and has a more stable radioactive tracer effect in the hydrogel formulation.

[0338] Although the embodiments of the present application have been shown and described, those of ordinary skill in the art can understand that various changes, modifications, substitutions, and variations can be made to these embodiments without departing from the principles and spirit of the present application. The scope of the present application is defined by the claims and their equivalents.

Claims

1. A radionuclide labeled polymer having the following structure: G-L1-L2-P; in, G is a group chelated with a radionuclide, which includes a chelating group and a radionuclide, wherein the chelating group chelates with the radionuclide; The L1 is a chemical bond or a connecting group represented by formula (I-1): （I-1）, In formula (I-1), x and y are independently selected from integers of 0 to 3; R1 is or ; R e for , substituted or unsubstituted C 6-10 Arylene, or substituted or unsubstituted C 5-10 heteroarylene; R d C 1-5 The alkylene or chemical bonds; R b is hydrogen, hydroxyl or C 1-5 alkyl; X is O or S; e represents the connection site between G and L1, i represents the connection site between R1 and R e , f represents the connection site between L1 and L2; The L2 is a group as shown in (II-2): (II-2), in, In formula (II-2), R2 and R3 are independently hydrogen, C 1-5 Alkyl, substituted or unsubstituted C 6-10 Aryl, or substituted or unsubstituted C 5-10 Heteroaryl, amide or thiol; f represents the connection site between L1 and L2, and h represents the connection site between L2 and P; u is 0, 1, 2, or 3; Each R 1-1 , R e-1 and R d-1 Independently for C 1-5 of alkylene; P is a temperature-sensitive structural unit, which is chitosan or a group as shown in formula (III-1), ， Wherein, in formula (III-1), a and c are independently selected from the range of 2-130; b is in the range of 10-80, and R is selected from hydrogen and C 1-5 alkyl.

2. The radionuclide-labeled polymer according to claim 1, characterized in that It satisfies one or more of the following conditions (1)-(5): (1) Each C 1-5 The alkylene group is methylene, , , or ; (2) Each C 6-10 Arylene is phenylene or naphthylene; (3) Each C 5-10 The heteroatoms in the heteroaryl group are independently selected from one or two of N, O and S, and the number of heteroatoms is independently 1, 2 or 3; (4) Each C 1-5 Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl or sec-butyl; (5) Each substituted or unsubstituted C 6-10 Arylene, or substituted or unsubstituted C 5-10 Heteroarylene, or substituted or unsubstituted C 6-10 Aryl, or substituted or unsubstituted C 5-10 The substituents in the heteroaryl group are independently C 1-3 alkyl, halogen, hydroxyl or cyano; The C 1-3 The alkyl group is methyl, ethyl or propyl; The halogen is F, Cl, Br or I.

3. The radionuclide-labeled polymer according to claim 2, characterized in that Each C 5-10 The heteroatom in the heteroaryl group is N or O, and the number of the heteroatoms is 1 or 2.

4. The radionuclide-labeled polymer according to claim 1, characterized in that It satisfies one or more of the following conditions (1)-(3): (1) L1 is a linking group represented by formula (I-2) or (I-3): （I-2）， （I-3）， Among them, in formula (I-2), R1 is or C 1-5 The alkylene group, i represents the e Connect; R e is substituted or unsubstituted C 6-10 Arylene, or substituted or unsubstituted C 5-10 Heteroarylene; wherein e represents a connection with G, and f represents a connection with L2; In formula (I-3), each n is independently selected from an integer in the range of 1 to 5, and each R b are independently hydrogen or hydroxy; (2) In formula (II-2), R2 and R3 are both methyl groups; (3) In formula (II-2), u is 0.

5. The radionuclide-labeled polymer according to claim 1, characterized in that In formula (III-1), a and c are independently integers in the range of 80-130, and b is an integer in the range of 30-60.

6. The radionuclide-labeled polymer according to any one of claims 1 to 5, characterized in that The radionuclide is 89 Zr; The chelating group is a chelating group formed by a bifunctional chelating agent.

7. The radionuclide-labeled polymer according to claim 6, characterized in that The chelating agent is DOTA, NOTA, NODA, NODAGA, DOTP, TETA, ATSM, PTSM, EDTA, EC, HBEDCC, DTPA, SBAD, BAPEN, DFO or TACN.

8. The radionuclide-labeled polymer according to claim 1, characterized in that It meets one or more of the following conditions: (1) The chelating group is , or ; (2) L1 is , or , e indicates connection with G, and f indicates connection with L2; (3) L2 is or , f represents the connection site between L1 and L2, and h represents the connection site between L2 and P; (4) P is or .

9. The radionuclide-labeled polymer according to claim 1, characterized in that: The polymer satisfies any one of the following options (1) to (3): Solution (1): The chelating group is ; The P is ; Wherein, R is a methyl group, a is an integer in the range of 75-110, and b is an integer in the range of 25-70; The L1 is , where e indicates connection with G, and f indicates connection with L2; The L2 is , f represents the connection site between L1 and L2, and h represents the connection site between L2 and P; Solution (2): The chelating group is ; The P is , wherein R is methyl, a is an integer in the range of 75-110, and b is an integer in the range of 25-70; The L1 is , where e indicates connection with G, and f indicates connection with L2; The L2 is , f represents the connection site between L1 and L2, and h represents the connection site between L2 and P; Solution (3): The chelating group is ; The P is , wherein R is methyl, a is an integer in the range of 75-110, and b is an integer in the range of 25-70; The L1 is , where e indicates connection with G, and f indicates connection with L2; The L2 is , f represents the connection site between L1 and L2, and h represents the connection site between L2 and P.

10. The radionuclide-labeled polymer according to claim 1, characterized in that It is any of the following structures: 、 or ； in, a and c are independently selected from the range of 2-130; b is in the range of 10-80, R is selected from hydrogen and C 1-6 alkyl.

11. The radionuclide-labeled polymer according to claim 10, characterized in that It is any of the following structures: 、 、 、 、 or 。 12. A polymer represented by formula (A): ; in, The definitions of L1, L2, and P are the same as those of L1, L2, and P in any one of claims 1 to 9; Q is a chelating group, and the chelating group is the same as the chelating group in any one of claims 1 to 9.

13. The polymer according to claim 12, characterized in that The polymer shown in formula (A) is any of the following structures: 、 or ， a and c are independently selected from the range of 2-130; b is in the range of 10-80, R is selected from hydrogen and C 1-6 alkyl.

14. The polymer according to claim 13, characterized in that The polymer represented by formula (A) is any of the following structures: 、 、 、 、 or 。 15. A method for preparing a radionuclide-labeled polymer, comprising the following steps: mixing and incubating a polymer represented by formula (A) as claimed in any one of claims 12 to 14 with a solution containing a radionuclide to obtain a radionuclide-labeled polymer.

16. The preparation method according to claim 15, characterized in that: The radionuclide-labeled polymer is as described in claim 10 or 11.

17. A gel composition comprising the radionuclide-labeled polymer according to any one of claims 1 to 11 and a hydrogel preparation.

18. The gel composition according to claim 17, characterized in that: It satisfies one or more of the following conditions (1)-(4): (1) The hydrogel preparation comprises a temperature-sensitive gelling agent and a gelling agent; (2) The hydrogel preparation further comprises a developer; (3) The mass volume ratio of the radionuclide labeled polymer to the hydrogel preparation is 0.01 mg / mL to 2 mg / mL; (4) The sol-gel transition temperature of the gel composition is 25°C-37°C.

19. The gel composition according to claim 18, characterized in that: The temperature-sensitive gelling agent and the radionuclide-labeled polymer have the same temperature-sensitive structural unit.

20. The gel composition according to claim 18, characterized in that: It satisfies one or more of the following conditions (1)-(3): (1) The temperature-sensitive gelling agent is one or more of acrylamide polymer, chitosan or polyethylene glycol block copolymer; (2) The gelling agent is one or more of sodium alginate, hydroxymethyl cellulose and CaCl2; (3) The developer is iohexol.

21. The gel composition according to claim 20, characterized in that: The temperature sensitive gelling agent is one or more of poloxamer, acrylamide polymer or chitosan.

22. The gel composition according to claim 21, characterized in that: Based on the volume of the hydrogel preparation, the hydrogel preparation comprises: 0.2-1g / mL iohexol, 0.02-0.1g / mL sodium alginate, 0.12-0.2g / mL poloxamer, 0.005-0.1 g / mL hydroxymethylcellulose or CaCl2, and water; and / or In the gel composition, the mass volume ratio of the radionuclide-labeled polymer to the hydrogel preparation is 0.01 mg / mL to 0.5 mg / mL.

23. The gel composition according to claim 22, characterized in that Based on the volume of the hydrogel preparation, the gel composition consists of 0.06 mg / mL of the radionuclide-labeled polymer, 0.4 g / mL of iohexol, 0.02 g / mL of sodium alginate, 0.14 g / mL of poloxamer, 0.005 g / mL of hydroxymethylcellulose and the balance of water.

24. A tracer comprising a polymer labeled with a radionuclide as claimed in any one of claims 1 to 11.

25. Use of a polymer labeled with a radionuclide according to any one of claims 1 to 11 in the preparation of a tracer, wherein the tracer is used to trace the gel composition according to any one of claims 17 to 23.

26. The use according to claim 25, characterized in that The tracer is a PET / CT imaging agent or a SPECT imaging agent.

27. A use of the gel composition according to any one of claims 17 to 23, wherein the use is use of the gel composition in preparing materials for in vivo vascular embolism, in vivo hemostasis, drug delivery or in vivo tissue repair.

28. A method for radiolabeling a hydrogel formulation, comprising: Providing a radionuclide-labeled polymer as claimed in any one of claims 1 to 11; The radionuclide-labeled polymer is added to the hydrogel formulation according to any one of claims 17-23.

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