Radionuclide-labeled polymer as well as preparation method and application thereof

Abstract

The invention discloses a radionuclide-labeled polymer as well as a preparation method and application thereof. Specifically, the invention provides a radionuclide labeled polymer which has the following structure: G-L1-L2-P, wherein G is a radionuclide-containing group and comprises a chelating group and a radionuclide, the chelating group is chelated with the radionuclide, and L1 and L2 are independently chemical bonds or linking groups; p is a temperature-sensitive structural unit. The radionuclide-labeled polymer can be used as a tracer of hydrogel, can be combined with the hydrogel under the condition that the property of the hydrogel is not changed, has good compatibility and stability, and further can be used for tracing implanted hydrogel and continuously tracking the position and state of the implanted hydrogel in real time.

Description

Technical Field

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

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

[0003] There is no method to continuously and in real time track the status and position of hydrogels after implantation, making the in vivo performance evaluation of implantable hydrogels extremely difficult. While existing angiography can image vascular embolism to a certain extent, the imaging duration is limited, making it impossible to continuously observe the status and position of implanted hydrogels, and thus unsuitable for imaging hydrogel materials with longer implantation cycles. Summary of the Invention

[0004] To address the imaging needs of existing hydrogel materials that are unable to adapt to longer implantation cycles, the present invention provides a radionuclide-labeled polymer, its preparation method, and its application. The radionuclide-labeled polymer provided by the present invention can be used as a tracer for hydrogels. It does not alter the properties of the hydrogel itself, exhibits good compatibility with the hydrogel, exhibits excellent binding stability, and maintains a high radioactivity purity both in vivo and in vitro for over 11 days. It can be used as a tracer for hydrogel implantation, enabling continuous, real-time tracking of the position and status of hydrogels after implantation, and can be used for in vivo performance evaluation of implantable hydrogels.

[0005] The present invention provides a radionuclide-labeled polymer having the following structure: G-L1-L2-P;

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

[0007] L1 and L2 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, L1 is a chemical bond or a linking group represented by formula (I-1):

[0013]

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

[0015] R1 is C 1-5 Alkylene, or chemical bonds;

[0016] R e for Substituted or unsubstituted C 6-10 Arylene, or substituted or unsubstituted C 5-10 heteroarylene;

[0017] R d C 1-5 Alkylene, or chemical bonds;

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

[0019] 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;

[0020] The L2 is a chemical bond, such as a group represented by formula (II-1) or (II-2):

[0021]

[0022] Wherein, in formula (II-1), z is 0, 1, 2 or 3;

[0023] 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, h represents the connection site between L2 and P;

[0024] u is 0, 1, 2, or 3;

[0025] Each R 1-1 、R e-1 and R d-1 Independent for C 1-5 of alkylene.

[0026] In one embodiment, P is a chitosan structural unit, an acrylamide structural unit, or a group represented by formula (III-1):

[0027]

[0028] 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.

[0029] In the present invention, in the compound represented by formula (I), each C 1-5 The alkylene group is methylene,

[0030]

[0031] In the present invention, each C 6-10 Arylene is phenylene or naphthylene, for example phenylene.

[0032] In the present invention, each C 5-10 Heteroaryl is preferably C 5-10 Heteroaryl, wherein the heteroatom is independently selected from one or two of N, O and S, and the number of heteroatoms is independently 1, 2 or 3, preferably the heteroatom is N or O, and the number of heteroatoms is 1 or 2, such as pyridine, pyrimidine or pyran.

[0033] In the present invention, each C 1-5 Alkyl is methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl or sec-butyl, for example methyl, ethyl or isopropyl.

[0034] In the present invention, 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; said C 1-3 The alkyl group is methyl, ethyl or propyl;

[0035] The halogen is F, Cl, Br or I.

[0036] The chelating group can be a conventional chelating group 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.

[0037] In one embodiment, the chelating group is

[0038] In one embodiment, L1 is a linking group represented by formula (I-2) or (I-3):

[0039]

[0040] Wherein, in formula (I-2), R1 is or C 1-5 alkylene, i represents connection with Re; 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;

[0041] In formula (I-3), each n is independently selected from an integer in the range of 0-10, R b is hydrogen or hydroxyl.

[0042] In one embodiment, in formula (I-2), R1 Or methylene i represents R e Linking site.

[0043] In one embodiment, in formula (I-2), R e is substituted or unsubstituted C 6-10 arylene groups, such as phenylene;

[0044] In one embodiment, in formula (I-2), R1 is methylene or And R e is phenyl, i represents R e connect.

[0045] In one embodiment, in formula (I-3), each n is independently 1, 2, 3, 4 or 5, R b It is a hydroxyl group.

[0046] In one solution, L1 is e indicates connection with G, and f indicates connection with L2.

[0047] In one embodiment, in formula (II-1), z is 2.

[0048] In one embodiment, in formula (II-2), R2 and R3 are both methyl.

[0049] In one embodiment, in formula (II-2), u is 0.

[0050] In one embodiment, in formula (II-2), R2 and R3 are both methyl, and u is 0.

[0051] In one solution, L2 is (For example )or

[0052] 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.

[0053] In one embodiment, the acrylamide structural unit includes a group represented by formula (III-2):

[0054]

[0055] In formula (III-2), R c is hydrogen, -CH-(CH3)2, -C-(CH3)3, or -(CH2) m -NH2, wherein m is an integer between 0 and 5, R a are independently -CH-(CH3)2, -C-(CH3)3, or -(CH2) m -NH2 is a divalent group formed by losing one hydrogen;

[0056] p or q is independently an integer in the range of 20-200.

[0057] In one embodiment, in formula (III-2), R c -CH-(CH3)2 or -(CH2) m -NH2, wherein m is 1, 2, 3, 4 or 5.

[0058] In one embodiment, in formula (III-2), R c It is -CH-(CH3)2 or -(CH2)2-NH2.

[0059] In one embodiment, in formula (III-2), R a -(CH2) m -NH2 loses hydrogen to form a divalent group, where m is an integer between 0 and 5, for example -(CH2)2-NH-.

[0060] In one embodiment, in formula (III-2), p is an integer in the range of 100-150, and q is an integer in the range of 20-40; for example, when R c When -CH-(CH3)2, p is 110-125, q is an integer in the range of 20-25 or R cWhen it is -(CH2)2-NH2, p is an integer in the range of 120-135, and q is an integer in the range of 25-35.

[0061] In one embodiment, P is

[0062] In one embodiment, the P is derived from a polymer represented by formula (IV):

[0063]

[0064] Wherein, in formula (IV), a, b, c and R are defined as above.

[0065] In one embodiment, in formula (IV), the average relative molecular weight of the polymer is 8000-13000 Da, such as 8600 Da-11500 Da, preferably 8600 Da or 11500 Da.

[0066] In some embodiments, the P is derived from a polymer represented by formula (V):

[0067]

[0068] In formula (V), R c is hydrogen, -CH-(CH3)2, -C-(CH3)3, or -(CH2) m -NH2, wherein 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.

[0069] In one embodiment, the polymer satisfies the following embodiments (1) to (5):

[0070] Solution (1):

[0071] The chelating group is

[0072] 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;

[0073] The L1 is Where e indicates connection with G, and f indicates connection with L2;

[0074] The L2 is f represents the connection site between L1 and L2, and h represents the connection site between L2 and P;

[0075] Solution (2):

[0076] The chelating group is

[0077] 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;

[0078] The L1 is Where e indicates connection with G, and f indicates connection with L2;

[0079] The L2 is f represents the connection site between L1 and L2, and h represents the connection site between L2 and P;

[0080] Solution (3):

[0081] The chelating group is

[0082] 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;

[0083] The L1 is Where e indicates connection with G, and f indicates connection with L2;

[0084] The L2 is f represents the connection site between L1 and L2, and h represents the connection site between L2 and P;

[0085] Solution (4):

[0086] The chelating group is The P is The L1 is Wherein, e represents connection with G, f represents connection with L2; L2 is a chemical bond;

[0087] Solution (5):

[0088] The chelating group is The P is The L1 is a chemical bond;

[0089] The L2 is a chemical bond.

[0090] In some embodiments of the present invention, the chemical bond is a covalent bond.

[0091] In a certain embodiment, the radionuclide-labeled polymer is any of the following structures:

[0092] Among them, a, b, c, p, q, R, R a and Rc As mentioned above, for example

[0093]

[0094]

[0095] In some embodiments, the radionuclide-labeled polymer is any of the following structures:

[0096]

[0097]

[0098]

[0099] The present invention also provides a polymer as shown in formula A:

[0100] Q-L1 - L2-P A;

[0101] Wherein, L1, L2, and P are as defined above, Q is a chelating group, and the chelating group is as defined above.

[0102] In one embodiment, the polymer as shown in formula A is any of the following structures:

[0103]

[0104]

[0105] Wherein a, b, c, p, q, R, Rc and Ra are as described above, for example

[0106]

[0107]

[0108] 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 described above with a solution containing a radionuclide to obtain the radionuclide-labeled polymer.

[0109] In one embodiment, in the preparation method, the radionuclide-labeled polymer is as described above.

[0110] In one embodiment, the preparation method comprises the following steps:

[0111] S1 provides an oxalic acid solution containing radioactive metal ions to obtain a mixed solution 1;

[0112] S2. The mixed solution 1 was incubated with Na2CO3 solution at room temperature to obtain a mixed solution 2;

[0113] S3. The mixed solution 2 is mixed with the polymer represented by formula (A) and a buffer solution, incubated, and the product is obtained after the reaction is completed;

[0114] Wherein, in step S1., the oxalic acid solution solvent is water;

[0115] In step S2., the Na2CO3 solution solvent is water.

[0116] Preferably, the preparation method further comprises:

[0117] S0. preparing the polymer to be labeled;

[0118] Wherein, step S0 includes the following steps:

[0119] S01. Synthesis of amino derivatives of temperature-sensitive polymers;

[0120] S02. Conducting a chelate reaction between the amino derivative and a chelating agent to generate a polymer represented by formula (A), wherein the polymer represented by formula (A) is a chelate of a polymer and a chelating group.

[0121] In one embodiment, S01 includes the following steps:

[0122] (1) reacting the amino-protected amino acid with the temperature-sensitive polymer in anhydrous dichloromethane in the presence of N,N'-dicyclohexylcarbodiimide and 4-dimethylaminopyridine to obtain a crude product;

[0123] (2) The crude product of step (1) is subjected to a deprotection reaction in anhydrous dichloromethane to obtain an amino derivative of a temperature-sensitive polymer.

[0124] In one embodiment, S02 includes the following steps:

[0125] (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;

[0126] Wherein, the amino-protected amino acid is a Boc-protected valine;

[0127] The deprotection reaction is to remove Boc protection;

[0128] The bifunctional chelating agent is one or more of DFO and DOTA.

[0129] The present invention also provides a gel composition comprising the radionuclide-labeled polymer and the hydrogel preparation as described above.

[0130] The hydrogel preparation of the present application may be a conventional hydrogel preparation in the art for in vivo vascular embolization, in vivo hemostasis, drug delivery, in vivo tissue filling or in vivo tissue repair, for example, preferably including a temperature-sensitive gelling agent and a gelling agent.

[0131] In a certain embodiment, the temperature-sensitive gelling agent is one or more of poloxamer, N-isopropylacrylamide polymer, chitosan or polyethylene glycol block copolymer.

[0132] In a certain embodiment, the gelling agent is one or more of sodium alginate, hydroxymethyl cellulose and CaCl2.

[0133] In one embodiment, the hydrogel formulation further comprises an imaging agent, such as iohexol.

[0134] In one embodiment, based on the total volume of the hydrogel formulation, 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 formulation comprises: 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 hydroxymethylcellulose or CaCl2, and water.

[0135] In one embodiment, the gelling agent is prepared by the following preparation method: dissolving sodium alginate and hydroxymethyl cellulose in an aqueous solution of iohexol, heating to dissolve, and cooling to obtain the gelling agent; in the aqueous solution of iohexol, the mass ratio of iohexol to water is (1-5):3, for example, 2:3.

[0136] In a certain embodiment, the hydrogel preparation includes one or more of sodium alginate, poloxamer and hydroxymethyl cellulose. Preferably, the hydrogel preparation consists of iohexol, sodium alginate, poloxamer, hydroxymethyl cellulose and water.

[0137] 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.

[0138] 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 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.

[0139] 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.

[0140] The present invention also provides a radionuclide-labeled polymer, which is prepared by the above-mentioned preparation method.

[0141] The present invention also provides a tracer comprising the radionuclide-labeled polymer as described above.

[0142] In one embodiment, the tracer is prepared by adding a polymer labeled with a radionuclide as described above to the hydrogel formulation as described above. The tracer is a PET / CT imaging agent or a SPECT imaging agent, so that the in vivo distribution of the gel composition can be observed by PET / CT imaging.

[0143] In one embodiment, the tracer is used to evaluate the embolic properties of the gel composition as described above.

[0144] The present invention also provides an application of the gel composition as described above, wherein the application is application of the gel composition in preparing materials for in vivo vascular embolism, in vivo hemostasis or in vivo tissue repair.

[0145] In a certain embodiment, the application is for preparing an embolic agent, wherein the embolic agent may be an embolic agent for transcatheter arterial chemoembolization for treating tumors.

[0146] The present invention also provides a tracing method for an implantable hydrogel, comprising the following steps:

[0147] (1) providing a hydrogel and adding a polymer labeled with a radionuclide as described above;

[0148] (2) implanted into a subject;

[0149] (3) Perform PET / CT imaging at multiple preset time points to check the imaging results and / or perform radioactivity counting.

[0150] The present application also provides a method for radiolabeling a hydrogel preparation, comprising:

[0151] providing a radionuclide-labeled polymer as described above;

[0152] The radionuclide-labeled polymer is added to the hydrogel formulation described above.

[0153] In one embodiment, the temperature-sensitive gelling agent and the radionuclide-labeled polymer have the same temperature-sensitive structural unit.

[0154] Preferably, when the temperature-sensitive gelling agent is poloxamer, P in the radionuclide-labeled polymer comprises a structure represented by formula (III-1);

[0155] When the temperature-sensitive gelling agent is an N-isopropylacrylamide polymer, P in the radionuclide-labeled polymer includes a structure represented by formula (III-2).

[0156] In one embodiment, the subject is a human, a primate, or a non-primate mammal.

[0157] In one embodiment, the subject is an experimental animal model.

[0158] In addition to the foregoing, when used in the specification and claims of this application, unless otherwise specifically indicated, the following terms have the following meanings:

[0159] As used herein, "temperature sensitivity" refers to the property of gelling from a liquid state, or solidifying from a liquid into a gel or solid state, as temperature increases. A "hydrogel preparation" refers to a water-soluble or hydrophilic polymer or a composition containing such a polymer, including a gel-like substance formed by chemical or physical crosslinking of the composition. The hydrogel preparation may include a temperature-sensitive polymer, such as a poloxamer or acrylamide polymer, or other temperature-sensitive polymers known in the art, and thus exhibits temperature-responsive properties, such as converting from a sol state to a gel state at a certain temperature. The hydrogel preparation may 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 may be any hydrogel material for medical, pharmaceutical, or cosmetic purposes.

[0160] As used herein, "phase transition temperature" refers to the temperature or temperature range at which a polymer undergoes a phase transition.

[0161] The term "polymer" refers to a molecule formed by the chemical combination of two or more oligomeric units. The chemical units are typically linked by covalent bonds. The two or more chemical units in a polymer can be identical, in which case the polymer is called a homopolymer. They can also be a combination of different units, in which case these polymers are called copolymers.

[0162] The term "chemical bond" refers to the general term for the strong interaction force between two or more adjacent atoms (or ions) in a pure molecule or crystal, generally ionic bond, covalent bond, and metallic bond.

[0163] The term "linking group" refers to the portion of an organic compound that has lost one or more atoms. It is a general term for groups of atoms and radicals that act as a linker. While a linker typically contains covalent bonds, it is not a chemical bond.

[0164] The term "heteroaryl" refers to an aromatic system containing 1, 2, 3 or 4 heteroatoms, wherein the heteroatoms are selected from N, O or S. Heteroaryl groups include, but are not limited to, thiazolyl, thienyl, pyridyl and pyrimidinyl.

[0165] Without violating the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain preferred embodiments of the present invention.

[0166] The reagents and raw materials used in the present invention are commercially available.

[0167] The beneficial effects of the present invention are as follows: the present invention provides a radionuclide-labeled polymer that can be used for mixing with a thermosensitive hydrogel without changing the physicochemical properties of the hydrogel itself, while still maintaining the hydrogel's original viscosity and thermosensitive curing properties. The radionuclide-labeled signal can exist stably in the body for a long time, maintaining a long half-life, and a continuous imaging period of more than 11 days. It can be used for in vivo tracking of embolic agents in the hydrogel, and the distribution of the hydrogel embolism in the body can be conveniently and continuously monitored without the need for multiple angiography.

[0168] The radionuclide-labeled polymer of the present invention is covalently linked to the polymer unit via a chelating group, which then chelates the radionuclide, resulting in better binding to the hydrogel preparation. When used for radiolabeling implantable hydrogels, it has better stability and in vivo imaging time than non-covalent linkages such as direct addition of zirconium salt chelation or hydrogen bonding. BRIEF DESCRIPTION OF THE DRAWINGS

[0169] Figure 1 For dog B01 89 Dynamic cross-sectional images of Zr-labeled poloxamer in vivo for 5 h;

[0170] Figure 2 For dog B01 89 Dynamic coronal images of Zr-labeled poloxamer in vivo for 5 h;

[0171] Figure 3 For dog B01 89 Dynamic cross-sectional images of Zr-labeled poloxamer 1d in vivo;

[0172] Figure 4 For dog B01 89 Dynamic coronal images of Zr-labeled poloxamer 1d in vivo;

[0173] Figure 5 For dog B01 89 Dynamic cross-sectional images of Zr-labeled poloxamer 5d in vivo;

[0174] Figure 6 For dog B01 89 Dynamic coronal images of Zr-labeled poloxamer 5d in vivo;

[0175] Figure 7 For dog B01 89 Dynamic cross-sectional images of Zr-labeled poloxamer 7d in vivo;

[0176] Figure 8 For dog B01 89 Dynamic coronal images of Zr-labeled poloxamer 7d in vivo;

[0177] Figure 9 For dog B01 89 Dynamic cross-sectional images of Zr-labeled poloxamer 11d in vivo;

[0178] Figure 10 For dog B01 89 Dynamic coronal images of Zr-labeled poloxamer 11d in vivo;

[0179] Figure 11 B04 dog 89 Dynamic cross-sectional images of Zr-labeled poloxamer in vivo for 5 h;

[0180] Figure 12 B04 dog 89 Dynamic coronal images of Zr-labeled poloxamer in vivo for 5 h;

[0181] Figure 13 B04 dog 89 Dynamic cross-sectional images of Zr-labeled poloxamer 1d in vivo;

[0182] Figure 14 B04 dog 89 Dynamic coronal images of Zr-labeled poloxamer 1d in vivo;

[0183] Figure 15 B04 dog 89 Dynamic cross-sectional images of Zr-labeled poloxamer 5d in vivo;

[0184] Figure 16 B04 dog 89 Dynamic coronal images of Zr-labeled poloxamer 5d in vivo;

[0185] Figure 17 B04 dog 89 Dynamic cross-sectional images of Zr-labeled poloxamer 7d in vivo;

[0186] Figure 18 B04 dog 89 Dynamic coronal images of Zr-labeled poloxamer 7d in vivo;

[0187] Figure 19 B04 dog 89 Dynamic cross-sectional images of Zr-labeled poloxamer 11d in vivo;

[0188] Figure 20 B04 dog 89 Dynamic coronal images of Zr-labeled poloxamer 11d in vivo;

[0189] Figure 21 for 89 Calibration curve of Zr activity meter detection value and γ counter detection value. DETAILED DESCRIPTION

[0190] The present invention is further illustrated by way of examples below, but the present invention is not limited to the scope of the examples. Experimental methods in the following examples where specific conditions are not specified were performed according to conventional methods and conditions, or selected according to the product specifications.

[0191] Example:

[0192] Material:

[0193] Polymer 1:

[0194]

[0195] Manufacturer: BASF CORPORATION (BASF), model P 407Geismar.

[0196] Polymer 2:

[0197]

[0198] Manufacturer: BASF CORPORATION (BASF), model P 188Geismar.

[0199] Chitosan: average relative molecular weight 5000Da, product number C434552, non-animal origin, purchased from Shanghai Aladdin Biochemical Technology Co., Ltd.

[0200] Iohexol: Manufacturer: Shanghai Star Pharmaceutical Co., Ltd.

[0201] Sodium alginate: Manufacturer: Qingdao Mingyue Alginate Tissue Engineering Materials Co., Ltd.

[0202] 89 Zr:

[0203] Manufacturer: PerkinElmer, Inc.

[0204] Purity and content: After unpacking, the total activity concentration was detected to be 19.54mCi / mL.

[0205] Prescription composition: 89-Zr is dissolved in 1 mol / L oxalic acid solution.

[0206] Storage conditions: Store at room temperature in a 2 mL V-bottom screw-cap glass bottle in a lead container. Protect from ionizing radiation.

[0207] N-isopropylacrylamide monomer: from Shanghai Aladdin, product number I106818.

[0208] N-(3-aminopropyl) methacrylamide monomer (APAM monomer): Shanghai Aladdin product number N129096.

[0209] Beagle dog (Beagle) animal source: Nanjing Chaimen Biotechnology Co., Ltd.

[0210] Other reagents are commercially available unless otherwise specified.

[0211] Preparation Example 1 Radiolabeled Polymer 89 Preparation of Zr-DFO-P1-1

[0212] 1.1 Synthesis of thermosensitive polymer and DFO chelate

[0213] (1) Polymer amino activation

[0214] 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 ultra-dry anhydrous DCM (dichloromethane) in a molar ratio of 1:2:2. The mixture was reacted at room temperature under argon for half an hour. 5.75 g of polymer 1 (P1-1) was added and the reaction was continued for 2 days. The DCM was concentrated by rotary evaporation and precipitated with glacial ether. The white precipitate was collected by filtration, redissolved with 3 mL of DCM, and precipitated with glacial ether again. The precipitation was repeated by filtration three times. The precipitate was redissolved in ultrapure water (heated) at a ratio of 1 mg:1 mL of ultrapure water, dialyzed for three days, and lyophilized to obtain 4.13 g of Boc-protected polymer P1-1-Val-Boc with a yield of 71.9%.

[0215]

[0216] (2) Removal of Boc protection

[0217] 4 g of polymer P1-1-Val-Boc was weighed and dissolved in 20 mL of DCM, placed in an ice-water bath, and 45 mL of trifluoroacetic acid was added under argon protection. The reaction was stirred for 2 hours, and the solvent was removed by rotary evaporation. The product was redissolved in chloroform, dried by rotary evaporation, dissolved in hot water, and dialyzed for three days. The product was lyophilized to obtain 3.2 g of the deprotected polymer P1-1-Val, with a yield of 80%.

[0218]

[0219] (3) Preparation of DFO chelate

[0220] 15.3 mg of polymer P1-1-Val was dissolved in 4 mL of 0.1 M NaHCO3 (sodium bicarbonate) solution, and p-SCN-Bn-DFO (CAS No.: 1222468-90-7) (2 mg; 200 μL DMSO) was added. The polymer P1-1-Val and p-SCN-Bn-DFO were slowly added dropwise at a molar reaction ratio of 1:2. The reaction was carried out at room temperature for 1 hour. After being spin-dried, the mixture was dissolved in hot water and dialyzed for three days. After freeze-drying, a total of 10.2 mg of polymer DFO-P1-1 was obtained with a purity of 99.3% and a yield of 66.7%.

[0221]

[0222] 1.2 89 Synthesis of Zr-labeled polymers

[0223]

[0224] (1) Prepare the marking solution

[0225] 89Oxalic acid solution of Zr: Zirconium [ 89 Zr] was dispersed in 1 M oxalic acid solution (pH < 4) (PerkinElmer, Inc);

[0226] To prepare 0.5M HEPES (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid) buffer: First, add 18 mL of ultrapure water to 20 mL of 1M HEPES buffer and check the pH. If the pH is < 7.1, adjust the pH with 1M NaOH. If the pH is > 7.3, adjust the pH with 1M H2SO4. Once the pH of the HEPES buffer is between 7.1 and 7.3, add additional volumes of ultrapure water to a total volume of 40 mL.

[0227] 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.

[0228] (2) 89 Zr labeling reaction

[0229] A. Accurately measure 84 μL 89 Add Zr oxalic acid solution to a glass reaction bottle;

[0230] B. Add 1 M oxalic acid solution to the glass reaction vial to make the total volume of the reaction vial 200 μL;

[0231] C. Use a pipette to accurately pipette 90 μL of 2M Na2CO3 solution into the reaction bottle and incubate at room temperature for 3 minutes;

[0232] D. While gently shaking the reaction bottle, dissolve 0.3 mL of 0.5 M 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 0.5 M HEPES to the reaction bottle. (Note: The pH of the reaction solution should be between 6.8 and 7.2 to obtain optimal labeling efficiency. Do not use metal syringe needles when preparing the solution.)

[0233] E. Incubate at room temperature for 1 hour while gently shaking the reaction bottle;

[0234] F. After the reaction is completed, use ultrafiltration centrifuge tube to purify the reaction product to remove free 89 Zr (washing solution: 10 mM pH 8.4 borate buffer solution; 500 μL*2 times).

[0235] Preparation Example 2 Radiolabeled Polymer 89 Preparation of Zr-DFO-P1-2

[0236]

[0237] Only polymer 1 (P1-1) in Preparation Example 1 was replaced by polymer 2 (P1-2), and the amount of polymer 2 was calculated based on the average relative molecular weight, and the reaction was carried out according to the steps of Preparation Example 1.

[0238] Preparation Example 3 Radiolabeled Polymer 89 Preparation of Zr-DOTA-P1-1

[0239]

[0240] The reaction was carried out according to the procedure of Preparation Example 1, except that p-SCN-Bn-DFO (CAS No.: 1222468-90-7) in Preparation Example 1 was replaced with p-SCN-Bn-DOTA (CAS No.: 127985-74-4).

[0241] Preparation Example 4 Radiolabeled Polymer 89 Preparation of Zr-DFO-P2

[0242] Chitosan was dissolved in 0.1 M acetic acid solution, p-SCN-Bn-DFO (2 mg; 200 μL DMSO) was added, chitosan and p-SCN-Bn-DFO were reacted in a molar ratio of 1:2 and slowly added dropwise at 37 ° C for 4 h, and then dissolved in hot water and dialyzed for three days after spin drying. The product DFO-P2 was obtained after freeze drying. Then the steps of step 1.2 in preparation example 1 were followed. 89 Radiolabeling of Zr 89 Zr-DFO-P2.

[0243] Preparation Example 5 Radiolabeled Polymer 89 Preparation of Zr-DOTA-P3-1

[0244] (1) Synthesis of Boc-protected N-isopropylacrylamide (NIPAM) polymer:

[0245] 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. 80.6 g of N-Boc-ethylenediamine was then added and reacted for 12 h. The mixture was then precipitated with 150 mL of diethyl ether, filtered, washed twice with diethyl ether, and dried to obtain pNIPAM-Boc with a yield of 83%.

[0246]

[0247] (2) Deprotection of Boc group:

[0248] 2.4 g of pNIPAM-Boc was dissolved in 50 mL of trifluoroacetic acid (8:2 volume ratio of trifluoroacetic acid to water) and stirred at room temperature for 2 h. The mixture was then rotary dried under vacuum and redissolved in water. The mixture was purified by Sephadex G25 column and freeze-dried to obtain pNIPAM with a yield of 90%.

[0249]

[0250] (3) DOTA activation: Dissolve 48 mg of DOTA in 2 mL of water and 9.0 mg of 1-ethyl-(3-dimethylaminopropyl)carbodiimide (EDC) in 0.5 mL of water. Mix the two aqueous solutions to form a reaction mixture. Adjust the pH of the reaction mixture to 5.0 with 0.1 M NaOH. Add 6.8 mg of NHS, stir in an ice bath, and react for 40 minutes to obtain DOTA-NHS.

[0251]

[0252] (4) Synthesis of pNIPAM-DOTA compound: 298 mg of pNIPAM (containing 0.03 mmol of amino groups) was dissolved in a mixed solvent containing 3 mmol of N,N-diisopropylethylamine (DIEA) and 6 mL of DMAC, heated to 45°C, and 0.03 mmol of DOTA-NHS was added. The mixture was reacted for 3 hours and purified by centrifugal filtration with a molecular weight cutoff of 3.5 kDa. After purification, the chelated ligand product DOTA-P3-1 was obtained by freeze-drying.

[0253]

[0254] (5) Refer to step 1.2 of Preparation Example 1 89 Radiolabeled with Zr to obtain 89 Zr-DOTA-P3-1.

[0255]

[0256] In this preparation example, the N-isopropylacrylamide polymer (the product of step (2) of Preparation Example 5) had an average molecular weight Mn of 17,400 g / mol as measured by size exclusion chromatography (SEC).

[0257] After DOTA was attached, the NMR spectra in D2O solution were recorded using a Varian AC400 NMR spectrometer (Varian NMR Systems, Palo Alto, CA). 1H-NMR, based on the hydrogen spectrum, showed that the molar ratio of the two repeating units, p:q, was approximately 0.85:0.15.

[0258] Preparation Example 6 Radiolabeled Polymer 89 Preparation of Zr-DFO-P3-2

[0259] (1) Add 3.37 g of N-(3-aminopropyl)methacrylamide monomer (APAM monomer) and 164 mg of azobisisobutyronitrile (AIBN) as an initiator, and synthesize by free radical reaction in an acetone-DMSO mixed solvent (V(acetone):V(DMSO)=15:1). Polymerize at 55°C under vacuum conditions for 24 hours, and filter and recover the precipitate to obtain a crude product. The crude product is dissolved in a small amount of methanol, and 15 times the volume of excess acetone is added to precipitate, and vacuum dried to obtain polyaminopropylmethacrylamide. The average molecular weight Mn is measured by size exclusion chromatography (SEC) and is 19200-21500 g / mol using a Synchropak CATSEC-300 column.

[0260] (2) p-SCN-Bn-DFO (CAS No. 1222468-90-7) (2 mg; 200 μL DMSO) was slowly added dropwise to 12 mg of polyaminopropyl methacrylamide, and sodium hydroxide was added to adjust the pH to 6.0. The reaction was allowed to proceed at room temperature for 1 h. After spin drying, the mixture was dissolved in water and dialyzed for three days. The molecular weight cutoff of the centrifugal filter membrane was 10 kDa. After freeze drying, a total of 9.7 mg of polymer was obtained, with a yield of 69.3%. The D2O solution was recorded using a Varian AC400 NMR spectrometer (Varian NMR Systems, Palo Alto, CA). 1 H-NMR, based on the hydrogen spectrum, showed that the molar ratio of the two repeating units, p:q, was approximately 0.82:0.18.

[0261] (3) Refer to step 1.2 of Preparation Example 1 89 Radiolabeled with Zr to obtain 89 Zr-DFO-P3-2.

[0262]

[0263] Preparation Example 7 Radiolabeled Polymer 89 Preparation of Zr-DFO-LP1-1

[0264] The p-SCN-Bn-DFO in Preparation Example 1 was replaced with compound D-1 (CAS: 1623757-39-0), and D-1 was reacted with P1-Val to prepare a DFO-coupled polymer, and then the reaction was carried out according to Step 1.2 of Preparation Example 1. 89 Zr marker.

[0265]

[0266] Preparation Example 8: Preparation 89 Zr-labeled hydrogel preparation

[0267] (1) Preparation 1

[0268] Prepare iohexol aqueous solution: accurately measure iohexol and water for injection in a mass ratio of 2g:3g, mix well, and prepare 40% iohexol solution.

[0269] Prepare a solution of sodium alginate and hydroxymethyl cellulose: weigh 2 g of sodium alginate and 0.5 g of hydroxymethyl cellulose (HPMC-100M), slowly add the sodium alginate and hydroxymethyl cellulose to 100 mL of 40% iohexol solution, slowly heat the room temperature to 35°C to promote the dissolution of the sodium alginate and hydroxymethyl cellulose, and let it cool to room temperature until they are completely dissolved.

[0270] Preparation of gel composition: Accurately weigh 14 g of poloxamer 407 and slowly add it to 100 mL of the prepared sodium alginate and hydroxymethyl cellulose solution. Let it stand at 4°C for 3-4 hours until completely dissolved to obtain Formulation 1. The sol-gel phase transition temperature is 26.5°C. At 37°C, it gels within 32 seconds and converts to a solid within 50 seconds. The viscosity is 0.9 Pa.s (25°C) and 1.2×10 3 Pa.S(30℃).

[0271] (2) Formulation 2: Embrace HES polyethylene glycol liquid embolic agent.

[0272] (3) Preparation 3: Peptide Pushen TM Temperature-sensitive liquid embolic agent (copolymer of N-isopropylacrylamide and N-n-propylacrylamide).

[0273] (4) Preparation of zirconium labeled gel composition:

[0274] Preparation 89 Zr polymer-labeled preparation: 1 mL of the 89 The Zr-labeled polymer solution (containing 0.6 mg of polymer) was mixed with 10 mL of Preparation 1, Preparation 2, or Preparation 3, stirred gently, and then refrigerated at 4-8°C.

[0275] (5) Preparation 89 Zr non-chelating agent labeled preparations: radioisotope 89 1 mL of the Zr oxalic acid solution was added to 10 mL of Preparation 1, Preparation 2, or Preparation 3 and mixed.

[0276] Test Example 1: Performance Test

[0277] 1.1 89 Radiochemical purity of Zr-labeled hydrogel preparations

[0278] A Mini-Scan thin-layer chromatography scanner (Eckert & Ziegler Radiopharma Inc., USA) was used with fast thin-layer chromatography paper (Agilent ITLC paper strips, 1 cm × 10 cm, USA) as the carrier. After spotting, the system was developed with 20 mM citric acid solution (pH 4.9-5.1). After the chromatography paper was dried, data was collected using a thin-layer chromatography scanner. After data collection was completed, images were obtained for data analysis.

[0279] The performance test results of the gel composition prepared by each polymer and hydrogel preparation according to the proportions of Preparation Example 8.(1) are as follows Table 1:

[0280] Table 1

[0281]

[0282] This study evaluated the 89 The radiochemical purity of the Zr-labeled hydrogel preparation and the labeling stability at different time points are shown in the table above. 89 The radiochemical purity of the Zr-labeled hydrogel preparations was maintained above 93% at different time stages, indicating that 89 The Zr-labeled hydrogel formulations showed good in vitro stability. 89 The radiochemical purity of Zr-labeled polymers used in hydrogel preparations was greater than 93%, meeting the experimental requirements, and the radiochemical purity was still greater than 93% after 19 days. 89 Zr oxalic acid solution, 89 Zr ions and the hydroxyl groups of sodium alginate undergo chelation and promote the cross-linking of sodium alginate to form a network structure. 89 Zr coating, to achieve 89 The chelate-free Zr coating has a high labeling efficiency at the initial labeling time of 1 day, but the radiochemical purity decreases with the extension of storage time, indicating that 89 The content of Zr-labeled sodium alginate decreases, and more 89 Zr detached from the label, and experimental group h showed poor stability. The polymer provided in this experimental example exhibits improved stability and is less susceptible to degradation due to the covalent chemical chelation of the chelating group with the polymer. This complexation of the radioactive metal element via the chelating group reduces the release of free radioactive elements and prolongs the radiolabeling and imaging time.

[0283] 1.2 89Sol-gel rheological properties of Zr-labeled gel compositions

[0284] The tests were performed on an AR-1000 constant stress rheometer (manufacturer: TA Instruments) with built-in temperature and gap calibration.

[0285] The rheometer is equipped with a cone / plate geometry (40 mm diameter, 4 ° cone angle). 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 an aqueous solvent trap to prevent sample moisture loss. During the test, the temperature of the peltier plate is controlled to within ± 0.1 ° C of the preset temperature. Before the test, the geometric gap is calibrated. After loading the excess sample, the geometry is lowered to the predetermined gap and the excess sample is removed. Before starting the test, the sample is allowed to equilibrate for 30 s. The data is processed using TA data analysis software.

[0286] Flow rheology was used to evaluate the viscosity of various hydrogel formulations. To determine how the hydrogels behave under increasing shear stress, steady-state flow experiments were performed at 25°C. The viscosity of the samples was measured over a shear stress range of 1 Pa to 100 Pa. The thermal responsiveness of the various hydrogel formulations was rheologically evaluated by oscillation measurements. A temperature sweep of 10°C-40°C was performed 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").

[0287] Therefore, gelation is considered to have occurred when G'>G". The temperature was increased at a rate of 1 °C / min, and the oscillation pressure and angular frequency remained unchanged.

[0288] The performance test results of the gel composition prepared by the polymer and hydrogel preparation prepared in the preparation example are shown in Table 2 below.

[0289] Table 2

[0290]

[0291] This indicates that the addition of a radiolabeled polymer has no significant effect on the viscosity and phase transition temperature of the liquid embolic agent, nor does it affect the rheological and temperature-sensitive properties of the hydrogel formulation itself. In particular, when the radiolabeled polymer and the temperature-sensitive material of the hydrogel formulation have identical or similar structural units, the radiolabeled polymer and the hydrogel formulation exhibit excellent compatibility, enabling more uniform and stable distribution within the hydrogel formulation, and minimizing the impact on the viscosity and phase transition temperature of the hydrogel formulation itself.

[0292] Test Example 2 In vivo animal test

[0293] This experiment observed the content of radioactive labeled polymer in the blood of animals, and used PET / CT tomography to collect signals within the window width range of 110keV±15%, and dynamically observed the radioactive labeled polymer prepared in Examples 1-7 of this application. 89 Dynamic distribution of Zr-labeled polymers within a certain period of time after implantation into Beagle dogs according to the protocol shown in Table 3.

[0294] Beagle dogs were placed under general anesthesia (1.5% sodium pentobarbital, 20 mg / kg), and the femoral artery was bluntly dissected. The femoral artery was punctured under direct vision, and a 5F sheath was placed. The femoral artery and sheath were secured with absorbable medical sutures. Under digital subtraction angiography (DSA) guidance, a 4F catheter was inserted through the femoral artery to the abdominal aorta. A Y-valve was connected to the distal end of the 4F catheter, and a 2.4F microcatheter was passed through the Y-valve and selectively inserted into the common hepatic artery. Gel was injected through the microcatheter, minimizing gel reflux. After embolization, the catheter and sheath were removed, the femoral artery puncture site was ligated, and the muscle and skin were sutured layer by layer.

[0295] The hydrogel prepared in the preparation example was used as an animal vascular embolic agent and implanted into Beagle dogs. The implantation scheme is shown in Table 3:

[0296] Table 3 Implantation plan

[0297] Implantation plan Hydrogel preparation Labeled polymers A Preparation 1 <![CDATA[ 89 Zr-DFO-P1-1]]> B Preparation 1 <![CDATA[ 89 Zr-DFO-P1-2]]> C Preparation 1 <![CDATA[ 89 Zr-DOTA-P1-1]]> D Preparation 1 <![CDATA[ 89 Zr-DFO-P2]]> E Preparation 2 <![CDATA[ 89 Zr-DFO-P1-1]]> F Preparation 2 <![CDATA[ 89 Zr-DFO-P1-2]]> G Preparation 2 <![CDATA[ 89 Zr-DOTA-P1-1]]> H Preparation 2 <![CDATA[ 89 Zr-DFO-P2]]> I Preparation 3 <h2 style=";text-align:left;direction:ltr"><![CDATA[ <h2 style=";text-align:left;direction:ltr"> 89 <h2 style=";text-align:left;direction:ltr"> Zr-DOTA-P3-1]]><h2 style=";text-align:left;direction:ltr"> J Preparation 3 <![CDATA[ 89 Zr-DFO-P3-2]]> K Preparation 1 <![CDATA[ 89 Zr-DFO-LP1-1]]> L Preparation 1 <![CDATA[ 89 Zr-DFO-P3-2]]> M Preparation 3 <![CDATA[ 89 Zr-DFO-P1-1]]> N Preparation 1 <![CDATA[ 89 Zr oxalate

[0298] 2.1 In blood 89 Zr detection

[0299] The dosage of the embolic agent was 1 mL of labeled gel per animal. The specific activity of the gel was 280 μCi / mL, and the dose given to each dog was approximately 280 μCi / animal. Blood was collected from the animal's cephalic vein in the forelimb using a sodium heparin anticoagulation negative pressure blood collection tube at 5 hours, 1 day, 3 days, 5 days, 7 days, and 11 days after hepatic artery embolization. The blood volume was approximately 2 mL. After blood collection, the blood samples were tested and analyzed using a WIZARD γ counter. 89 The calibration curve of the Zr activity meter detection value (μCi) and the γ counter detection value (CPM) is used for the conversion of activity and CPM. The calibration curve is shown in Figure 21 .

[0300] The blood volume was estimated based on the dog's systemic circulating blood volume and blood leakage count results, animal dosage and body weight data. 89 Zr dose and calculate blood 89The radioactivity of Zr as a percentage of the total dose. For Beagle dogs with a circulating blood volume of 85 mL / kg, the total blood volume was calculated based on the animal's body weight. The total blood exposure was then calculated based on the total blood volume. The percentage of blood exposure was calculated by comparing this with the total dose after attenuation. The percentage of blood exposure as a percentage of the total dose = body weight (kg) x 85 mL / kg x blood test value (μCi / g) / administered dose. Blood test values ​​were corrected to the time of administration.

[0301] Table 4 Animal blood 89 Zr radioactivity

[0302]

[0303] Table 5 Animal blood 89 Zr radioactivity percentage

[0304]

[0305] As shown in Table 4 and Table 5, on the first day of embolization, the blood of dogs B01-B14 89 The percentage of Zr radioactivity in the total dose was less than 7% and then gradually decreased. Seven days after administration, the radioactivity of Zr in the blood of dogs B01-B13 was 100%. 89 The radioactivity of Zr accounts for approximately 0.69% to 2% of the total administered dose.

[0306] Overall, blood 89 The radioactivity of Zr is relatively low, indicating that the gel formed by the test substance mainly remains in the embolization site in vivo, and also proves that the test substance has good stability after labeling and long-term placement. 89 After the hydrogel preparation of Zr-labeled polymer, no free Zr was found in the blood. 89 Abnormal increase or decrease of Zr indicates better stability in the animal body and no obvious free state is observed.

[0307] The polymer provided in this test example is 89 Zr is chemically chelated with the polymer through the chelate group, which is better than 89 The Zr oxalate co-labeled preparation has better stability, is not easily degraded, and reduces the release of free radioactive elements.

[0308] Dog B14 implanted with gel 89 Zr is directly mixed and coated without chemical chelation of thermosensitive polymer. The radioactivity in the blood within 5h-11d after implantation is significantly higher than that of other implants including 89 The radioactive signal in the blood was high within 1-5 days, indicating that radioactive elements were constantly released into the blood circulation. After 5 days, most of the embolization sites were 89Zr has entered the blood, no more 89 Zr escapes, so the radioactivity in the blood decreases after 7 days. The radioactivity entering the blood circulation within 11 days accounts for 60% of the total dose, indicating that direct addition of free 89 Zr, the radioactive label has poor stability and is easily dissociated from the embolization site. The radioactive label cannot stably track the position of the hydrogel for a long time.

[0309] In implantation 89 In animals treated with Zr-labeled polymer hydrogel formulations, polymers of different structures as radioactive markers in the hydrogel formulations all showed lower levels of blood radioactivity, demonstrating good stability. In particular, when the hydrogel formulation and the radioactively labeled polymer contained the same thermosensitive polymer units, the hydrogel formulation and the radioactively labeled polymer had more similar viscosities and phase transition temperatures, enabling better compatibility and tight binding, and the radioactive labeling exhibited excellent stability. For example, when a radioactively labeled polymer containing an alkyl ether structural unit was added to formulation 1 or 2, 89 Zr-DFO-P1-1 or 89 When Zr-DFO-P1-2 was added, the total proportion of radioactive counts in the blood during the test period was significantly lower than that added to preparation 3; radiolabeled polymers containing acrylamide structural units 89 When Zr-DFO-P3-2 was added to Formulation 3 containing an acrylamide polymer as a temperature-sensitive component, it also exhibited significantly better stability than when it was added to Formulation 1.

[0310] Compared with animals B01 and B11, the only difference in the radioactive polymer linker in the preparation is that the tracing effect of animal B01 is better, indicating that 89 Zr-DFO-P1-1 ratio 89 Zr-DFO-LP1-1 has a better effect, which may be due to 89 The chain length of the linking group in Zr-DFO-LP1-1 is affected.

[0311] 2.2 In vivo dynamic imaging studies

[0312] Collection time: 5h, 1d, 5d, 7d, 11d, and 18d after embolization.

[0313] Acquisition method: Nuclear medicine tomography + positioning CT. After the animal is placed under general anesthesia and strapped into the Discovery VCT instrument, a full-body CT scan is performed first to assist in tissue and organ identification, followed by a full-body nuclear medicine signal scan from the head to the bottom of the lower limbs.

[0314] PET-CT imaging data processing: Coronal and cross-sectional images of the PET-CT imaging results of dogs B01-B14 were saved. Regions of interest (ROIs) were delineated (major organs with nuclear medicine signals) based on the coronal and cross-sectional images and nuclear medicine images. The corresponding areas were circled, and the percentage of the counts in the delineated areas to the total body counts was recorded.

[0315] The results are as follows Figures 1-20 and as shown in Table 6.

[0316] Figure 1 、 Figure 3 、 Figure 5 、 Figure 7 、 Figure 9 These are the cross-sectional images of dog B01 at 0.5 h, 1 day, 5 days, 7 days, and 11 days after implantation of the radiolabeled hydrogel preparation; Figure 11 、 Figure 13 、 Figure 15 、 Figure 17 、 Figure 19 These are transverse images of dog B04 taken 0.5 hours, 1 day, 5 days, 7 days, and 11 days after implantation of the radiolabeled hydrogel. In each transverse image, the upper left corner shows the CT localization map, the upper right corner shows the nuclear medicine image of the localization location, the lower left corner shows the cross-sectional nuclear medicine image of the enriched region, and the lower right corner shows the whole-body nuclear medicine signal image (this image does not change with the CT localization map). Each region is located in the same location—the liver.

[0317] Figure 2 、 Figure 4 、 Figure 6 、 Figure 8 、 Figure 10 These are the coronal images of dog B01 at 0.5 h, 1 day, 5 days, 7 days, and 11 days after implantation of the radiolabeled hydrogel preparation; Figure 12 、 Figure 14 、 Figure 16 、 Figure 18 、 Figure 20 These are transverse images of dog B04 taken 0.5 hours, 1 day, 5 days, 7 days, and 11 days after implantation of the radiolabeled hydrogel. Coronal images: The first left corner of the image shows a CT localization map, the second left shows a nuclear medicine image of the localization location, the third left shows a coronal nuclear medicine image of the enriched region, and the fourth left shows a whole-body nuclear medicine image. All regions are located in the same location: the liver.

[0318] The images show that in the animal (B01) implanted with supplementation protocol A, the radioactive signal was primarily distributed in the liver, with smaller amounts in the intestines and bladder, starting 5 hours after implantation. From 1 to 11 days after implantation, the radioactive signal remained primarily in the liver, with minimal amounts in other organs. Compared to B01, in the animal (B04) implanted with supplementation protocol D, the radioactive signal was primarily distributed in the liver, but significantly more was seen in the lungs, intestines, kidneys, and brain.

[0319] Table 6 below records the average percentage of the total body counts of the five dogs, namely, the liver, lung, intestine, kidney, and brain, which are the five areas with higher counts.

[0320] Table 6 Percentage of organ counts in the total body count of each animal (%)

[0321]

[0322] As shown in Table 6, on the day of administration, the liver accounted for over 80% of the drug in each animal, indicating that the test substance embolized in the liver on the day of administration, and the vast majority of the radioactive signals were at the embolization site, indicating high labeling efficiency and almost no degradation. During the 1-5 days after administration, the liver accounted for a reduced percentage of the test substance. This may be because some of the test substance embolized near the hepatic portal. The radiolabeled polymer that failed to fully bind to the hydrogel during the administration phase was washed away by the bloodstream and migrated to other tissues such as the intestine, lung, and kidney, where it was enriched. Therefore, slight distribution was also seen in the intestine, lung, and kidney on days 5-7, and the radioactive count percentage showed a slightly increasing trend. However, the liver count percentage still accounted for over 80%, still significantly marking the embolization site, and did not affect the assessment of the embolic status. From the 11th to the 18th day after intervention, and from the 5th to the 18th day after administration, the proportion of the test substance in the liver gradually increased to 100%, and the signals of the intestinal wall and other tissue 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. Only the liver showed a signal, indicating that the test substance remained in the embolization site in the body, and 89 The Zr-labeled polymers showed good long-term stability in vivo.

[0323] Throughout the experimental period, 89 Zr-labeled polymers are mainly enriched in the liver and the average proportion is between 80% and 100%, indicating that most of the radioactive nuclides are tightly bound to the implant and do not detach. They can be significantly distinguished from the free weak signals and are excellent tracers for implantable hydrogels.

[0324] Specifically, in animal B01, implanted with supplementation regimen A, the radioactivity signal in the liver was significantly higher starting at 5 hours. The liver maintained high radioactivity intensity throughout the monitoring period of 1 to 18 days, exceeding 80% or 84%. Radioactivity signals in other organs, such as the lungs, intestines, kidneys, and brain, were very low. This indicates that in animal B01, the radiolabeled polymer was tightly bound to Formulation 1 and embolized in the liver, demonstrating good stability of the embolic formulation. Furthermore, the radiolabeled polymer maintained good stability, with minimal detachment from the embolization site. Animals B02 and B05, implanted with supplementation regimen B and E, also maintained over 80% of the radioactivity count in the liver throughout the monitoring period. In contrast, animal B04, implanted with supplementation regimen D, showed relatively low radioactivity signal concentrations in the liver, initially reaching 80.9%. This suggests that after implantation with supplementation regimen D, a small amount of radioactivity was released into the bloodstream and circulated to other organs. However, by day 18, the radioactivity in organs other than the liver had been metabolized in animal B04. It can be seen that the polymer based on polymer P1 and the linking group 89 Compared with chitosan, Zr-DFO-P1-1 directly reacts with the amino group to form a polymer 89 The Zr-DFO-P2 structure has stronger stability and a more stable radioactive tracer effect in hydrogel preparations.

[0325] Although the embodiments of the present application have been shown and described, those skilled in the art will appreciate that various changes, modifications, substitutions, and variations may be made to the embodiments without departing from the principles and intent of the present application, and that 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 comprises a chelating group and a radionuclide, wherein 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.

2. The radionuclide-labeled polymer according to claim 1, characterized in that The L1 is a chemical bond or a connecting group represented by formula (I-1): In formula (I-1), x and y are independently selected from integers of 0 to 3; R1 is C 1-5 Alkylene, or chemical bonds; R e for Substituted or unsubstituted C 6-10 Arylene, or substituted or unsubstituted C 5-10 heteroarylene; R d C 1-5 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 of L1 and L2; and / or, The L2 is a chemical bond, such as a group represented by formula (II-1) or (II-2): Wherein, in formula (II-1), z is 0, 1, 2 or 3; 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, 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 Independent for C 1-5 of alkylene.

3. The radionuclide-labeled polymer according to claim 2, characterized in that It satisfies one or more of the following conditions (1)-(5): (1) Each C 1-5 The alkylene group is methylene, (2) Each C 6-10 Arylene is phenylene or naphthylene; (3) Each C 5-10 Heteroaryl is preferably C 5-10 Heteroaryl, wherein the heteroatom is independently selected from one or two of N, O and S, and the number of heteroatoms is independently 1, 2 or 3, preferably the heteroatom is N or O, and the number of heteroatoms is 1 or 2; (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.

4. The radionuclide-labeled polymer according to claim 2, characterized in that It satisfies one or more of the following conditions (1)-(4): (1) L1 is a linking group represented by formula (I-2) or (I-3): Wherein, in formula (I-2), R1 is or C 1-5 The alkylene group, i represents the alkylene group with R 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 0-10, and each R b are independently hydrogen or hydroxy; (2) In formula (II-1), z is 2; (3) In formula (II-2), R2 and R3 are both methyl; (4) In formula (II-2), u is 0.

5. The radionuclide-labeled polymer according to any one of claims 1 to 4, characterized in that The P is one or more of a chitosan structural unit, an acrylamide structural unit, 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.

6. The radionuclide-labeled polymer according to claim 5, characterized in that Satisfy at least one of the following conditions (1)-(2): (1) 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; (2) The acrylamide structural unit includes a group represented by formula (III-2): In formula (III-2), R c is hydrogen, -CH-(CH3)2, -C-(CH3)3, or -(CH2) m -NH2, wherein m is an integer between 0 and 5, R a is -CH-(CH3)2, -C-(CH3)3, or -(CH2) m -NH2 is a divalent group formed by losing one hydrogen; p or q is independently an integer in the range of 20-200.

7. The radionuclide-labeled polymer according to any one of claims 1 to 4, characterized in that The radionuclide is 89 Zr; The chelating group is a chelating group formed by a bifunctional chelating agent.

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

9. 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 (2) L1 is e indicates connection with G, f indicates connection with L2; (3) L2 is f represents the connection site between L1 and L2, and h represents the connection site between L2 and P; (4)P is 10. The radionuclide-labeled polymer according to claim 1, characterized in that The polymer satisfies any one of the following schemes (1) to (5): 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 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 (3): 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 (4): The chelating group is The P is The L1 is Wherein, e represents connection with G, f represents connection with L2; L2 is a chemical bond; Solution (5): The chelating group is The P is The L1 is a chemical bond; The L2 is a chemical bond.

11. The radionuclide-labeled polymer according to claim 1, characterized in that It is any of the following structures: 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; R c is hydrogen, -CH-(CH3)2, -C-(CH3)3, or -(CH2) m -NH2, wherein m is an integer between 0 and 5, R a is -CH-(CH3)2, -C-(CH3)3, or -(CH2) m -NH2 is a divalent group formed by losing one hydrogen; p or q is independently an integer in the range of 20-200.

12. The radionuclide-labeled polymer according to claim 11, characterized in that It is any of the following structures:

13. 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 11. Q is a chelating group, and the chelating group is the same as the definition of the chelating group in any one of claims 1 to 11.

14. The polymer according to claim 13, characterized in that The polymer represented by formula (A) is any of the following structures: 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; R c is hydrogen, -CH-(CH3)2, -C-(CH3)3, or -(CH2) m -NH2, wherein m is an integer between 0 and 5, R a is -CH-(CH3)2, -C-(CH3)3, or -(CH2) m -NH2 is a divalent group formed by losing a hydrogen. p or q is independently an integer in the range of 20-200.

15. The polymer according to claim 14, characterized in that The polymer represented by formula (A) is any of the following structures:

16. A method for preparing a radionuclide-labeled polymer, comprising the steps of: mixing and incubating a polymer represented by formula (A) according to any one of claims 13 to 15 with a solution containing a radionuclide to obtain a radionuclide-labeled polymer.

17. The preparation method according to claim 16, characterized in that: The radionuclide-labeled polymer is as described in any one of claims 1 to 12.

18. A gel composition comprising the radionuclide-labeled polymer according to any one of claims 1 to 12 and a hydrogel preparation.

19. The gel composition according to claim 18, characterized in that It satisfies one or more of the following conditions (1)-(4): (1) The hydrogel preparation includes a temperature-sensitive gelling agent and a gelling agent; (2) The hydrogel formulation 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.

20. The gel composition according to claim 19, wherein It satisfies one or more of the following conditions (1)-(3): (1) The temperature-sensitive gelling agent is one or more of poloxamer, 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, wherein 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.

22. The gel composition according to claim 21, 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 hydroxymethyl cellulose, and the balance of water.

23. A tracer comprising the radionuclide-labeled polymer according to any one of claims 1 to 12.

24. Use of the radionuclide-labeled polymer according to any one of claims 1 to 12 in the preparation of a tracer, wherein the tracer is used to trace the gel composition according to any one of claims 18 to 22.

25. The use according to claim 24, characterized in that The tracer is a PET / CT imaging agent or a SPECT imaging agent.

26. Use of the gel composition according to any one of claims 18 to 22, wherein the use is use of the gel composition in preparing materials for in vivo vascular embolization, in vivo hemostasis, drug delivery or in vivo tissue repair.

27. A method for radiolabeling a hydrogel formulation, comprising: Providing a radionuclide-labeled polymer according to any one of claims 1 to 12; The radionuclide-labeled polymer is added to the hydrogel formulation described in the gel composition according to any one of claims 18 to 22.

Images