Drug for diagnosing diabetic nephropathy and use thereof

Abstract

The present invention provides a compound represented by general formula (I) or a pharmaceutically acceptable salt, precursor, or solvate thereof, and a use thereof in the preparation of a drug for diagnosing diabetic nephropathy. General formula (I), wherein X is selected from carbon or nitrogen; R1 is selected from hydrogen or alkyl; R2 is one or more substituents on the benzene ring or the pyridine ring, and R2 is independently selected from one or more of hydrogen, halogen, hydroxyl, cyano, nitro, amino, C1-C6 alkyl, C3-C6 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C6 cycloalkoxy, C1-C6 alkylamino, C3-C6 cycloalkylamino, halo C1-C6 alkyl, halo C3-C6 cycloalkyl, halo C1-C6 alkoxy, halo C3-C6 cycloalkoxy, halo C1-C6 alkylamino, halo C3-C6 cycloalkylamino, C6-C8 aryl, or C5-C8 heteroaryl, and in these substituents represented by R2, at least one atom or the entire substituent is replaced by a radionuclide. The drug and method for diagnosing diabetic nephropathy according to the present invention relate to non-invasive diagnosis technology and have the advantages of causing less pain to a subject and having less contraindications.

Description

Diabetic nephropathy diagnostic drugs and their applications Technical Field

[0001] The present invention relates to a drug for diagnosing diabetic nephropathy, in particular to a radioactive therapeutic drug, and specifically to use of a triazolotetrahydropyridine ring compound in diagnosing diabetic nephropathy and a method for diagnosing diabetic nephropathy. Background Art

[0002] Diabetic nephropathy (DKD) is a common complication of diabetes and one of the important causes of end-stage renal failure. Early diagnosis of DKD is particularly important. Generally speaking, it is believed that the estimated glomerular filtration rate (eGFR) <30mL·min -1 1.73m -2 Renal damage is irreversible. Early drug intervention can delay the onset of organic kidney disease. Timely detection of early glomerular, tubular, and interstitial lesions is conducive to the early diagnosis and treatment of DKD. A retrospective study of 121,395 subjects showed that early diagnosis of DKD can reduce the risk of subjects progressing to end-stage renal disease by 80% (Chinese Journal of Internal Medicine. 2021. 60(6): 522-532.). Because the management and treatment of DKD are quite different from non-diabetic nephropathy (NDKD), clinicians should accurately diagnose DKD and identify NDKD based on clinical manifestations and auxiliary examinations, so as to provide timely and correct treatment for subjects. Currently, the diagnosis of diabetic nephropathy (DKD) is often based on albuminuria and the presence of diabetic retinopathy. However, albuminuria lacks specificity for DKD, and some DKD subjects may present with negative urine protein in the early stages, with only a decreased GFR. Therefore, diabetic subjects with albuminuria or decreased GFR may have DKD, NDKD, or a combination of both. Therefore, before diagnosing DKD, the presence of diabetes and NDKD should be ruled out. Currently, renal biopsy is the gold standard for diagnosing DKD and excluding NDKD. If the subject has no contraindications to renal biopsy, renal biopsy is recommended for pathological diagnosis of DKD (Expert Consensus on Prevention and Treatment of Diabetic Nephropathy, 2014 edition).

[0003] Japanese Patent Application Laid-Open No. 2010-256132A discloses a method for detecting the progression of diabetic nephropathy, a diagnostic kit for determining the progression of diabetic nephropathy, and a substance and method that serve as an indicator of the progression of diabetic nephropathy. In this patent application, body fluid collected from a subject is reacted with a lectin to measure the amount of sugar chains with affinity for the lectin. The measured sugar chain amount is then used to detect the progression of diabetic nephropathy.

[0004] International patent application WO2020071517 discloses biomarkers, miRNA-125b-5p and / or miRNA-181b-5p, that can specifically diagnose diabetic nephropathy. In this diagnosis, increased expression of miRNA-125b-5p and / or decreased expression of miRNA-181b-5p in the blood are used to diagnose diabetic nephropathy.

[0005] Summary of the Invention

[0006] Technical Problems to be Solved by the Invention

[0007] As mentioned above, renal biopsy is currently the most effective method for diagnosing diabetic nephropathy. However, renal biopsy is an invasive procedure and must exclude contraindications such as a significant bleeding tendency, severe hypertension, mental illness, or uncooperative patients, as well as patients with solitary or small kidneys. Therefore, renal biopsy has certain limitations.

[0008] In view of the limitations of the prior art, the object of the present invention is to provide a non-invasive and contraindication-free drug and method for diagnosing diabetic nephropathy.

[0009] Technical solutions to technical issues

[0010] The present inventors have sought compounds that specifically accumulate in the kidneys of subjects with diabetic nephropathy. Among the compounds described in International Publication No. WO2014 / 152604A1, which discloses affinity for the P2X7 receptor (P2X7R), the inventors discovered that the compound represented by the following general formula (I) specifically accumulates in the kidneys of subjects or models with diabetic nephropathy. The present invention utilizes radionuclide labeling of the compound and positron emission tomography (PET) or single-photon emission computed tomography (SPECT) imaging techniques to localize and quantify the accumulation of the compound in the kidneys, thereby diagnosing diabetic nephropathy.

[0011] Specifically, the present invention includes the following technical solutions.

[0012] The compound represented by general formula (I) or a pharmaceutically acceptable salt, precursor, or solvate thereof,

[0013] wherein X is selected from carbon (C) or nitrogen (N);

[0014] R 1 is selected from hydrogen or alkyl;

[0015] R 2is one or more substituents on the benzene ring or pyridine ring, R 2 R is independently selected from one or more of hydrogen, halogen, hydroxy, cyano, nitro, amino, C1-C6 alkyl, C3-C6 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C6 cycloalkyloxy, C1-C6 alkylamino, C3-C6 cycloalkylamino, C1-C6 hydroxyalkyl, C3-C6 hydroxycycloalkyl, halo-C1-C6 alkyl, halo-C3-C6 cycloalkyl, halo-C1-C6 alkoxy, halo-C3-C6 cycloalkyloxy, halo-C1-C6 alkylamino, halo-C3-C6 cycloalkylamino, C6-C8 aryl or C5-C8 heteroaryl, and R 2 At least one atom in these substituents or the entire substituent is substituted by a radioactive nuclide.

[0016] In the compound of general formula (I), R 1 Preferred are hydrogen, C1-C6 alkyl or C3-C6 cycloalkyl.

[0017] In the compound of general formula (I), R 2 It is one or more substituents on the benzene ring or pyridine ring, for example, it can be 1, 2, 3, 4 or 5 substituents. When there are 2 or more substituents, these substituents may be the same or different.

[0018] R 2 Further preferred are one or more of hydrogen, halogen, hydroxyl, nitro, amino, C1-C4 alkyl, C1-C4 alkoxy, C1-C4 alkylamino, halogenated C1-C4 alkyl, halogenated C1-C4 alkoxy or halogenated C1-C4 alkylamino, and at least one of these substituents has an atom or the entire group substituted with a radionuclide.

[0019] In the compound of general formula (I), the radionuclide is preferably 18 F. 11 C. 131 I. 123 I. 124 I. 125 One or more of I.

[0020] The compound of general formula (I) is preferably selected from any one of the following compounds:

[0021] (S)-(3-Fluoro-2-trifluoromethylpyridin-4-yl)(6-methyl-1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone,

[0022] (S)-(2-Fluoro-6-methylpyridin-4-yl)(6-methyl-1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone,

[0023] (4-iodophenyl)(1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone,

[0024] (3-iodo-4-methoxyphenyl)(1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone,

[0025] (2-iodo-4-nitro)(1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone,

[0026] (2-Fluoro-4-(tri-n-butyltin)phenyl)(1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone,

[0027] (2-Fluoro-4-iodophenyl)(1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone.

[0028] The precursor compound is selected from:

[0029] (S)-(3-chloro-2-trifluoromethylpyridin-4-yl)(6-methyl-1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone,

[0030] (S)-(2-chloro-6-methylpyridin-4-yl)(6-methyl-1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone,

[0031] (1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)(4-tri-n-butyltinphenyl)methanone,

[0032] (4-methoxy-3-(tri-n-butyltinyl)phenyl)(1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone,

[0033] (4-nitro-2-(tri-n-butyltin)phenyl)(1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone,

[0034] (2-Fluoro-4-(tri-n-butyltinyl)phenyl)(1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone.

[0035] The pharmaceutically acceptable salts of the compounds of the present invention are addition salts of inorganic or organic acids, wherein the inorganic acid is selected from hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, and nitric acid, and the organic acid is selected from acetic acid, tartaric acid, salicylic acid, methanesulfonic acid, succinic acid, lemon segment, malic acid, lactic acid, and fumaric acid.

[0036] The solvate of the compound of the present invention is preferably a hydrate.

[0037] The present invention also provides use of any of the above compounds or pharmaceutically acceptable salts, precursors, or solvates thereof in the preparation of diagnostic drugs for diabetic nephropathy.

[0038] The present invention also provides a radioactive diagnostic kit for diagnosing diabetic nephropathy, wherein the radioactive diagnostic reagent comprises a compound having a structure represented by the above general formula (I) or a pharmaceutically acceptable salt, precursor or solvate thereof.

[0039] In the diagnostic kit of the present invention, the pharmaceutically acceptable salt is an addition salt of an inorganic acid or an organic acid. The inorganic acid is selected from hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, and nitric acid, and the organic acid is selected from acetic acid, tartaric acid, salicylic acid, methanesulfonic acid, succinic acid, lemon segment, malic acid, lactic acid, and fumaric acid.

[0040] In the diagnostic kit of the present invention, the solvate is a hydrate.

[0041] The present invention also provides a method for diagnosing diabetic nephropathy, comprising:

[0042] Formulating any one of the above compounds, or a pharmaceutically acceptable salt, precursor, or solvate thereof, into a physiologically acceptable solution;

[0043] introducing the solution into a subject;

[0044] Imaging of the subjects' kidneys;

[0045] The images are analyzed and a diagnosis is made based on the results of the analysis.

[0046] Among the above diagnostic methods, imaging by PET or SPECT is preferred.

[0047] In the above diagnostic method, the physiologically acceptable solution can be introduced into the subject by, for example, injection.

[0048] Beneficial technical effects

[0049] The present invention provides a radioactive diagnostic drug, its use in diagnosing diabetic nephropathy, and a method for diagnosing diabetic nephropathy. By intravenously injecting the radioactive diagnostic drug and then performing PET or SPECT imaging, non-invasive diagnosis of diabetic nephropathy can be achieved. Compared with existing invasive renal puncture, this method offers the advantages of less pain for the patient and fewer contraindications. BRIEF DESCRIPTION OF THE DRAWINGS

[0050] Figure 1 shows the normal mice and diabetic nephropathy model mice 18 F-KIDJI-003 ex vivo autoradiography images.

[0051] Figure 2 shows the normal mice and diabetic nephropathy model mice 18 Quantitative analysis results of F-KIDJI-003 in vivo autoradiography.

[0052] FIG3 shows a radiolabeled compound of the present invention. 18 F-KIDJI-004, 131 I-KIDJI-007、 131 I-KIDJI-008、 131 I-KIDJI-009 and 131 In vivo SPECT imaging of I-KIDJI-010 in the kidneys of db / db model mice and normal mice.

[0053] Figure 4 shows the kidneys of normal mice, diabetic nephropathy mice, heterozygous and homozygous 18 F-KIDJI-003 in vitro autoradiography images.

[0054] FIG5 shows the results of P2X7R immunostaining experiments in the kidneys of normal mice, heterozygous diabetic nephropathy mice, and homozygous diabetic nephropathy mice. DETAILED DESCRIPTION

[0055] Features, characteristics, compounds, chemical moieties or groups described in conjunction with an embodiment or example of the invention are to be understood as being applicable to any other embodiment or example described herein, unless incompatible therewith. All features disclosed in this specification (including claims, abstract and drawings) and / or all steps of any method or process disclosed thereby may be combined in any way, except for at least some mutually exclusive combinations of such features and / or steps. The invention is not limited to the details of any embodiment. The invention extends to any new feature or any new combination of features disclosed in this specification (including claims, abstract and drawings), or to any new step or any new combination of steps of any method or process disclosed thereby.

[0056] Imaging Isotopes and Imaging:

[0057] Diagnostic techniques in nuclear medicine use radioactive tracers that emit gamma rays from within the body. These tracers are typically short-lived isotopes linked to a chemical compound that allow detailed examination of specific physiological processes. They can be administered by injection, inhalation, or orally. The first involves the detection of single photons by a gamma camera, which can view organs from many different angles. The camera creates an image based on the points where the radiation was emitted; this image is enhanced by a computer and viewed by the doctor on a monitor for signs of abnormalities.

[0058] Positron emission tomography (PET) is a precise and complex technique that uses isotopes produced by cyclotrons. Positron-emitting radionuclides are usually introduced by injection and accumulate in the target tissue. When the radionuclide decays, it emits a positron, which quickly combines with a nearby electron, resulting in the simultaneous emission of two identifiable gamma rays in opposite directions. These are detected by the PET camera and indicate their source very accurately. The most important clinical role of PET is in oncology, where it is used to detect 18-fluorodeoxyglucose (F-18DG) in the presence of fluorine. 18 F]FDG) as a tracer because it has been shown to be the most accurate non-invasive method for detecting and evaluating most cancers. It also works well for cardiac and brain imaging.

[0059] Many medical diagnostic procedures, including PET and SPECT, utilize radiolabeled compounds, which are well known in the art. PET and SPECT are very sensitive techniques and require small amounts of radiolabeled compounds (called tracers). Radiolabeled compounds are transported, accumulated, and transformed in the body in a similar manner to corresponding non-radiolabeled compounds. Tracers or probes can use radionuclides useful for PET imaging, such as 11 C. 13 N. 15 O. 18 F. 64 Cu and124 I is radiolabeled, or radionuclides useful for SPECT imaging such as 99 Tc, 77 Br, 61 Cu, 153 Gd, 123 I. 125 I. 131 I and 32 These are non-limiting examples of the term "radionuclide" (also known as radioisotope, imaging isotope) as used herein.

[0060] About radioactive halogens, isotopes 123 I has a half-life of 13 hours and a gamma energy of 159 keV, so this isotope is usually used 18 F (half-life 2 hours) labels the ligand to be used for diagnostic purposes. Other imaging isotopes that can be used include 131 I. 77 Br and 76 Br.

[0061] Those skilled in the art are familiar with various ways of detecting labeled compounds for imaging purposes. For example, radiolabeled compounds can be detected using positron emission tomography (PET) or single photon emission computed tomography (SPECT). The label incorporated into the compound may depend on the desired detection method. Those skilled in the art are familiar with positron emitting atoms such as 18 PET detection of F. Those skilled in the art are familiar with photon-emitting atoms such as 123 I or 99 SPECT detection of Tc.

[0062] Radioactive diagnostic or detection agents should have sufficiently high radioactivity and radioactivity concentration to ensure reliable diagnosis and detection. The required radioactivity level can be obtained by the methods provided herein for preparing the compounds.

[0063] Typically, in the first step of an imaging method, a labeled compound is introduced into a tissue or subject in a detectable amount. The compound is typically part of a pharmaceutical composition and is administered to the tissue or subject by methods well known to those skilled in the art. Typically, administration is by intravenous injection.

[0064] In other embodiments of the invention, a labeled compound is introduced into a subject in a detectable amount, and the labeled compound is detected non-invasively after sufficient time has passed for the compound to accumulate in the kidney.

[0065] A detectable amount is the amount of labeled compound necessary for detection by the selected detection method. The amount of labeled compound to be introduced into a subject to provide detectable expression can be readily determined by one skilled in the art. For example, the amount of labeled compound administered to a subject can be increased until the compound is detected using the selected detection method. Radionuclides are introduced into the compound to provide for detection of the compound.

[0066] The required amount of time can be readily determined by introducing a detectable amount of a labeled compound into a subject and then detecting the labeled compound at various times after administration.

[0067] Radiolabeled compounds can be administered to a subject by a general or local administration route. For example, the labeled compound can be administered to the subject so that the compound is delivered to the whole body. Alternatively, the labeled compound can be administered to a specific organ or tissue of interest. For example, it is desirable to locate and quantitatively analyze the compound level labeled in the kidney to diagnose or track the progress of, for example, diabetic nephropathy in the subject.

[0068] By replacing one or more atoms (e.g., hydrogen or alkyl, halogen, etc.) in the P2X7 receptor (P2X7R) affinity compound described in prior art WO2014 / 152604A1 with a radionuclide, one or more radionuclides can be incorporated into the compounds disclosed herein. Known techniques can be used for incorporation of radionuclides. For example, these techniques can be based on nucleophilic or electrophilic reactions of suitable precursors. 18 F fluorination is reviewed, for example, in Medicinal Chemistry Approaches to Personalized Medicine (Lackey, Roth ed.), Chapter 12 (Wiley-VCH, ISBN 978-3-527-33394-3).

[0069] definition

[0070] In the examples of the present invention, unless otherwise specified, all instruments, raw materials, experimental animals, and chemical reagents are commercially available products familiar to those skilled in the art; in the examples of the present invention, unless otherwise specified, the technical means used are conventional means familiar to those skilled in the art. The progress of the reaction of the present invention can be monitored using conventional monitoring methods in the art (e.g., TLC, HPLC, LCMS, or NMR), and the reaction endpoint is generally determined to be the disappearance of the reaction substrate.

[0071] Experimental methods in which specific conditions are not specified in the examples of the specification are generally carried out in accordance with conventional conditions in the art or the conditions recommended by the manufacturer. In the present invention, unless otherwise specified, "above", "below", and "within" are intended to include the number itself. When "comprising" or "including" are used to describe an embodiment herein, other similar embodiments described as "consisting of..." and / or "consisting essentially of..." are also provided. When "consisting essentially of..." is used to describe an embodiment herein, other similar embodiments described as "consisting of..." are also provided. The term "and / or" as used in phrases such as "A and / or B" herein is intended to include both A and B; A or B; A (alone); and B (alone). Similarly, the term "and / or" as used in phrases such as "A, B and / or C" is intended to include each of the following embodiments: A, B and C; A, B or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0072] In the present invention, the term "C1-C6 alkyl" refers to a straight or branched chain alkyl group having 1 to 6 carbon atoms in the chain. For example, methyl (Me), ethyl (Et), n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl (tBu), n-pentyl, isopentyl (2-methylbutyl), neopentyl (2,2-dimethylpropyl), n-hexyl, 2-methylpentyl, 3-methylpentyl, 2,3-dimethylbutanyl, 2,4-dimethylbutanyl, 3,3-dimethylbutanyl, 2-ethylbutanyl, etc., as well as groups that are considered to be equivalent to any of the foregoing examples based on the common skills in the art and the teachings provided herein. Similarly, the term "C1-C4 alkyl" refers to a straight or branched chain alkyl group having 1 to 4 carbon atoms in the chain.

[0073] The term "C3-C6 cycloalkyl" refers to a saturated monocyclic or polycyclic carbocyclic ring having 3 to 6 carbon atoms, such as cyclopropyl, 1-methylcyclopropyl, cyclobutyl, cyclopentyl, 1-methylcyclobutyl, cyclohexyl, 1,3-dimethylcyclobutyl, 1-methylcyclopentyl, and groups that would be considered equivalent to any of the foregoing examples based on ordinary skill in the art and the teachings provided herein.

[0074] The term "C2-C6 alkenyl" refers to a group having 2 to 6 carbon atoms and containing at least one carbon-carbon double bond, which may be straight or branched. The group may contain multiple double bonds, and the orientation of each double bond may be independently E or Z. The alkenyl group is preferably a 1-alkenyl group. Exemplary alkenyl groups include, but are not limited to, ethenyl, propenyl, butenyl, pentenyl, hexenyl, and groups that would be considered equivalent to any of the foregoing examples based on the ordinary skill in the art and the teachings provided herein. The group may be a terminal group or a bridging group.

[0075] The term "C2-C6 alkynyl" refers to a group having 2 to 6 carbon atoms and containing at least one carbon-carbon triple bond, and may be straight or branched. The group may contain multiple triple bonds. The alkynyl group is preferably a 1-alkynyl group. Exemplary alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, hexynyl, and groups that would be considered equivalent to any of the foregoing examples based on ordinary skill in the art and the teachings provided herein. The group may be a terminal group or a bridging group.

[0076] The term "C1-C6 alkoxy" refers to a C1-C6 alkyl-O- group, wherein C1-C6 alkyl is as defined above. The group may be a terminal group or a bridging group. Specifically, C1-C6 alkoxy includes methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, n-pentoxy, isopentoxy (e.g., 2-methylbutoxy, 2-methoxybutyl), neopentoxy (e.g., 2,2-dimethylpropoxy), cyclopentoxy, n-hexyloxy, 2-methylpentoxy, 2-methoxypentyl, 3-methylpentoxy, 3-methoxypentyl, 2,3-dimethylbutoxy, 2,4-dimethylbutoxy, 3,3-dimethylbutoxy, 2,3-dimethoxybutyl, 2,4-dimethoxybutyl, 3,3-dimethoxybutanyl, 2-ethylbutoxy, 2-ethoxybutanyl, and the like, as well as groups that would be considered equivalent to any of the foregoing examples based on ordinary skill in the art and the teachings provided herein.

[0077] The term "C1-C6 alkylamino" refers to an NH2-alkyl(C1-C6) group, wherein alkyl(C1-C6) is as defined herein for C1-C6 alkyl. The group may be a terminal group or a bridging group. If the group is a terminal group, the group is bonded to the rest of the molecule via the alkyl group.

[0078] The term "halogen" denotes fluorine, chlorine, bromine or iodine.

[0079] The term "halo C1-C6 alkyl" refers to a C1-C6 alkyl group in which a halogen is optionally substituted for hydrogen. C1-C6 alkyl is as defined above. Examples include, but are not limited to, trifluoromethyl (CF3), difluoromethyl (CF2H), monofluoromethyl (CH2F), pentafluoroethyl (CF2CF3), tetrafluoroethyl (CHFCF3), monofluoroethyl (CH2CH2F), trifluoroethyl (CH2CF3), tetrafluorotrifluoromethylethyl (-CF(CF3)2), and groups that would be considered equivalent to any of the foregoing examples according to the ordinary skill in the art and the teachings provided herein.

[0080] The term "C1-C6 haloalkoxy" refers to a C1-C6 alkoxy group optionally replacing hydrogen with a halogen. C1-C6 alkoxy is defined as above. Examples of haloalkoxy groups include trifluoromethoxy (OCF3), difluoromethoxy (OCF2H), monofluoromethoxy (OCH2F), monofluoroethoxy (OCH2CH2F), pentafluoroethoxy (OCF2CF3), tetrafluoroethoxy (OCHFCF3), trifluoroethoxy (OCH2CF3), tetrafluorotrifluoromethylethoxy (-OCF(CF3)2), and groups that are considered to be equivalent to any one of the foregoing examples according to the ordinary skill in the art and the teachings provided herein.

[0081] The term "C1-C6 haloalkylamino" refers to a C1-C6 alkylamino group optionally substituted with halogen for hydrogen. C1-C6 alkylamino is as defined above.

[0082] The term "C6-C8 aryl" refers to an aromatic carbocyclic ring having a ring structure in which all ring atoms are carbon atoms, preferably each ring has 6 to 8 carbon atoms, and examples of aryl groups include phenyl, etc. The group may be a terminal group or a bridging group.

[0083] The term "C5-C8 heteroaryl" refers to a group containing an aromatic ring (preferably a 5- or 6-membered aromatic ring) having one or more heteroatoms as ring atoms in the aromatic ring, with the remaining ring atoms being carbon atoms. Suitable heteroatoms include nitrogen, oxygen and sulfur. Examples of heteroaryl groups include thiophene, furan, imidazole, thiazole, isothiazole, [2,3-b]thiophene, isoindolizine, pyrrole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, tetrazole, indole, isoindole, 1H-indazole, purine, quinoline, isoquinoline, phthalazine, naphthyridine, quinoxaline, cinnoline, carbazole, phenanthridine, acridine, phenazine, isothiazole, phenothiazine, oxazole, isoxazole, furazan, phenoxazine, 2-, 3-, or 4-pyridyl, 2-, 3-, 4-, 5-, or 8-quinolyl, 1-, 3-, 4-, or 5-isoquinolyl, 1-, 2-, or 3-indolyl, and 2- or 3-thienyl. The group may be a terminal group or a bridging group.

[0084] The term "substituted" refers to a specified group or part with one or more substituents. The term "optionally substituted" refers to a specific group that is unsubstituted or substituted with one or more substituents. If the term "substituted" is used to describe a structural system, it refers to that substitution occurs at any valence-allowed position on the system. Where a specified part or group is not explicitly indicated to be optionally substituted or substituted with any specified substituent, it is understood that such part or group is intended to represent that it is unsubstituted. The terms "one or more", "one or more" mentioned in the substituent refer to from one substituent to the highest possible substitution number chemically, i.e., an atom is replaced with its corresponding radioisotope until all atoms are replaced.

[0085] The term "diagnosis" refers to the act of identifying a disease from its signs and symptoms, and in the present invention, specifically the analysis of biomarkers indicative of the disease.

[0086] The term "subject" includes humans and non-human animals. Non-human animals include all vertebrates, e.g., mammals and non-mammals, such as non-human primates, sheep, dogs, cows, chickens, amphibians, and reptiles. Except where noted, the terms "patient" or "subject" are used interchangeably herein.

[0087] The term "P2X7R" refers to the P2X7 receptor.

[0088] The term "pharmaceutically acceptable" refers to products or compounds approved by the Chinese drug regulatory authorities or listed in the Chinese Pharmacopoeia or other generally recognized pharmacopoeias for use in animals including humans.

[0089] The "precursor" of the present invention is a chemical substance in the previous stage of the production of the compound of formula (I). This chemical substance can produce a radiolabeled compound of formula (I) through a one-step radiolabeling chemical reaction.

[0090] The compounds of general formula (I) can form addition salts with suitable non-toxic organic or inorganic acids. Examples of acid addition salts include salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid and nitric acid, and salts derived from organic acids such as acetic acid, tartaric acid, salicylic acid, methanesulfonic acid, succinic acid, citric acid, malic acid, lactic acid, fumaric acid, etc. These salts can be prepared from the compounds of general formula (I) according to known salt-forming methods.

[0091] The compounds of the present invention can be recovered from the reaction mixture and purified in a conventional manner. Isomers such as enantiomers can be obtained in a conventional manner, for example, by stepwise synthesis from corresponding asymmetrically substituted starting materials. If necessary, protection of any active group can be carried out in any suitable step. The protecting group is preferably a protecting group conventionally used in the prior art, and conventional methods can be used to introduce and remove them. For example, when the amino group is protected by Boc, the Boc can be removed by conventional methods under acidic conditions.

[0092] The compounds of the present invention can be synthesized according to known techniques, and the present invention is described in detail below with reference to examples.

[0093] Example 1: Preparation method of the precursor compound of the present invention

[0094] The precursor compound of the present invention was prepared according to the following scheme:

[0095] Example 1-1:

[0096] Synthesis of (S)-(3-chloro-2-(trifluoromethyl)pyridin-4-yl)(6-methyl-1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone (precursor of KIDJI-003)

[0097] Step 1. Synthesis of compound 2

[0098] 2-Chloropyrimidine (10.0 g, 1.0 eq) was placed in a 100 mL three-necked flask under N2 protection and maintained at 30°C or less. Hydrazine hydrate (20 mL, 4.7 eq) was slowly added dropwise and stirred at room temperature for 16 h. TLC confirmed the complete reaction (pure ethyl acetate, with the product becoming more polar). The mixture was concentrated to dryness under reduced pressure, and then blended with n-hexane (50 mL) for 2 h. The mixture was filtered and the filter cake was washed with n-hexane (20 mL). The solid was concentrated to dryness and pumped dry to obtain 7.7 g of a white solid with an 80% yield.

[0099] Step 2. Synthesis of compound 3

[0100] Compound 2 (5 g, 1.0 eq) was placed in a 250 ml three-necked flask under N2 protection. A 50% aqueous glacial acetic acid solution (100 mL) was added and the temperature was lowered to below 0°C. A solution of sodium nitrite (6.3 g, 2.0 eq) in water (25 mL) was slowly added dropwise. The temperature was controlled at T≤5°C and the addition was completed in half an hour. The mixture was reacted at 0°C for about 2 hours. After TLC monitoring, the reaction of the raw materials was completed. Solid sodium carbonate was slowly added to adjust the pH to about 8. The mixture was extracted three times with toluene. The organic layer was collected, dried, filtered, and used directly in the next reaction without concentration.

[0101] Step 3. Synthesis of compound 4

[0102] Compound 3 (0.7 g, 1.0 eq) and tert-butyl (S)-2-methyl-4-oxopiperidine-1-carboxylate (0.95 g, 0.77 eq) were placed in a 100 mL three-necked flask. Toluene (10 mL) was added under nitrogen protection. The temperature was raised to 70°C. Tetrahydropyrrole (0.316 g, 0.77 mmol) was injected via syringe and the temperature was raised to 100°C. The reaction was allowed to proceed for 16 h, with the formation of new spots observed by TLC monitoring. The product was cooled to room temperature, concentrated under reduced pressure, and purified by column chromatography (PET / EA = 1:10).

[0103] Step 4. Synthesis of compound 5

[0104] Compound 4 (0.5 g, 1.0 eq) was placed in a 100 mL three-necked flask under N2 protection. Anhydrous dichloromethane (10 mL) was added, followed by sodium bicarbonate (0.11 g, 1.0 eq). m-CPBA (0.33 g, 1.0 eq) was slowly added, and the mixture was stirred at room temperature for 2 hours. TLC was used to monitor the complete reaction of the starting materials. After the reaction was complete, 1N aqueous sodium hydroxide solution (10 mL) was added, stirred for 20 minutes, and extracted with dichloromethane. The organic phase was collected, dried, concentrated under reduced pressure, and used directly in the next reaction.

[0105] Step 5. Synthesis of compound 6

[0106] Compound 5 (1 g, 1.0 eq) was placed in a 100 mL single-necked bottle under N2 protection, and TFA (2.4 g, 10 eq) was added. The mixture was stirred at room temperature for 16 h. After the reaction of the raw material was completed, TLC was monitored. The mixture was concentrated under reduced pressure to remove TFA, and dichloromethane was added. A 10% aqueous sodium carbonate solution was added to adjust the pH to about 8. The mixture was extracted with dichloromethane three times. The organic phase was collected, dried, filtered, concentrated under reduced pressure, and purified by column chromatography (DCM / MeOH = 15:1).

[0107] Step 6.

[0108] Synthesis of (S)-(3-chloro-2-(trifluoromethyl)pyridin-4-yl)(6-methyl-1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone (Compound 7; KIDJI-003 precursor)

[0109] Compound 6 (60 g, 1.0 eq) was placed in a 100 mL three-necked flask under N2 protection. Chloronicotinic acid (50 mg, 1.1 eq), HATU (91 mg, 1.1 eq), DIPEA (0.15 ml, 4.0 eq), DMF (5 ml), and DCM (10 ml) were added and stirred at room temperature for 16 h. LCMS analysis indicated the formation of the product. The mixture was washed with saturated brine (20 ml) and extracted with dichloromethane. The organic phase was collected, concentrated under reduced pressure, and purified by column chromatography; the mixture was then spin-dried to obtain a white solid.

[0110] 1 H NMR(500MHz,Chloroform-d)δ8.84-8.79(m,2H),8.65-8.57(m,1H),7.37-7.31(m,2H),5.79-5.72,5.5 5(m,1H),4.57,3.97-3.93(m,1H),4.38-4.30(m,1H),3.44-3.16(m,2H),1.32-1.29,1.18-1.16(m,3H).

[0111] Example 1-2: Preparation of KIDJI-004 precursor

[0112] According to the method of Example 1-1, chloronicotinic acid was replaced with 2-chloro-6-methylisonicotinic acid to prepare (S)-(2-chloro-6-methylpyridin-4-yl)(6-methyl-1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone (KIDJI-004 precursor).

[0113] Example 1-3:

[0114] According to the method of Example 1-1, (S)-2-methyl-4-oxopiperidine-1-carboxylic acid tert-butyl ester was replaced by N-tert-butyloxycarbonyl-4-piperidone; and chloronicotinic acid was replaced by 3-n-butyltinbenzoic acid to prepare (1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)(4-tri-n-butyltin)phenyl)methanone (KIDJI-007 precursor).

[0115] Example 1-4:

[0116] According to the method of Example 1-1, (S)-2-methyl-4-oxopiperidine-1-carboxylic acid tert-butyl ester was replaced by N-tert-butyloxycarbonyl-4-piperidone; and chloronicotinic acid was replaced by 3-n-butyltin-4-methoxybenzoic acid to prepare (4-methoxy-3-(tri-n-butyltin)phenyl)(1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone (KIDJI-008 precursor).

[0117] Example 1-5:

[0118] According to the method of Example 1-1, (S)-2-methyl-4-oxopiperidine-1-carboxylic acid tert-butyl ester was replaced by N-tert-butyloxycarbonyl-4-piperidone; and chloronicotinic acid was replaced by 2-n-butyltin-4-nitrobenzoic acid to prepare (4-nitro-2-(tri-n-butyltin)phenyl)(1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone (KIDJI-009 precursor).

[0119] Examples 1-6:

[0120] According to a method similar to that of Example 1-1, (S)-2-methyl-4-oxopiperidine-1-carboxylic acid tert-butyl ester was replaced with N-tert-butyloxycarbonyl-4-piperidone; and chloronicotinic acid was replaced with 2-fluoro-4-n-butyltinylbenzoic acid to prepare (2-fluoro-4-(tri-n-butyltinyl)phenyl)(1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone (KIDJI-010 precursor).

[0121] Example 2: Labeled synthesis

[0122] Labeling Synthesis Example 2-1

[0123] 18 F-labeled (S)-(3-fluoro-2-(trifluoromethyl)pyridin-4-yl)(6-methyl-1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone ( 18 Labeled synthesis of F-KIDJI-003)

[0124] The following route was used to prepare (S)-(3-chloro-2-(trifluoromethyl)pyridin-4-yl)(6-methyl-1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone (Compound 7, KIDJI-003 precursor) as the labeled precursor. 18F-labeled (S)-(3-fluoro-2-(trifluoromethyl)pyridin-4-yl)(6-methyl-1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone ( 18 F-KIDJI-003).

[0125] Weigh amino polyether (kryptofix) 222 (5 mg) and K2CO3 (3 mg) separately and place them in a 1.5 mL EP tube. Add pure water (100 μL) and acetonitrile (400 μL) to prepare kryptofix-222 / K2CO3 solution. Take the kryptofix-222 / K2CO3 solution and elute it through a QMA column. 18 Place F- in a 3 mL conical bottom fluorination reaction tube and heat at 135°C with nitrogen bubbling for 3-5 minutes. Add 300 μL of anhydrous acetonitrile to the fluorination reaction tube and heat at 135°C with nitrogen bubbling for 2 minutes. Repeat three times to fully remove water and dry.

[0126] Compound 7 (2 mg) was dissolved in anhydrous DMSO (200 μL), and 4 μL of KF (potassium fluoride) solution (1.5 mg / mL DMSO) was added. The mixture was added to the reaction tube and stirred at 135°C for 10 minutes. After the reaction was completed, the entire reaction solution was separated and purified by HPLC. HPLC used a reverse C18 column (Waters Atlantis T3 10×250mm, 5um); the mobile phase was CH3CN / H2O (0.1% TFA) = 36:64; the flow rate was 2.0 mL / min. The components with a retention time of 37-38 minutes were collected, and the obtained fractions of the labeled compound were transferred to a 20 mL rotary evaporation flask and concentrated under reduced pressure to dryness in the rotary evaporation flask. Physiological saline (containing 2.5% (V / V) Tween 80 and 2.5% (V / V) ascorbic acid solution) was added to prepare 18 F-KIDJI003 injection. Radiochemical purity >95%. Molar activity 61-87 GBq / μmol.

[0127] 1 H NMR(500MHz,Chloroform-d)δ8.95-8.91(m,2H),8.68-8.64(m,1H),7.60-7.56(m,1H),7.49-7.44(m,1H), 5.82,5.65-5.62(m,1H),4.69,4.19-4.16(m,1H),4.58-4.40(m,1H),3.58-3.37(m,2H),1.42-1.28(m,3H).

[0128] Labeling Synthesis Example 2-2

[0129] 18 F-labeled (S)-(2-fluoro-6-methylpyridin-4-yl)(6-methyl-1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone ( 18 Labeled synthesis of F-KIDJI-004)

[0130] This product was prepared by following a similar labeling procedure as in Example 2-1, using (S)-(2-fluoro-6-methylpyridin-4-yl)(6-methyl-1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone as the precursor. Radiochemical purity was >95%. Molar activity was 58-90 GBq / μmol.

[0131] Labeling Synthesis Example 2-3:

[0132] 131 I-labeled (4-iodophenyl)(1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone ( 131 I-KIDJI-007) labeled synthesis

[0133] The following route was used to prepare (1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)(4-tri-n-butyltin)phenyl)methanone as the labeled precursor. 131 I-KIDJI007.

[0134] Place 65 μL of phosphate buffer (0.3 M, pH = 5.5) in a 1.5 mL low adsorption reaction tube, add 20 μL of labeled precursor (1 mg / mL methanol), and add 5 μL 131I-NaI (200 μCi, Xinke Pharmaceutical) was added to 5 μL of chloramine T (0.4 mg / mL H2O). The reaction was vortexed at room temperature for 3 min, and then quenched with 100 μL of Na2S2O5 (2.0 mg / mL H2O). Purification was performed by high-performance liquid chromatography (HPLC column, CAPCELLPAK C18UG120 (φ6.0 mm × 150 mm; Shiseido, Tokyo, Japan); mobile phase, CH3CN / H2O, 10 / 90 to 100 / 0, v / v; flow rate, 1.0 mL / min). The radioactive product was collected at the appropriate time points. This process was repeated until sufficient radioactive drug (1.2-3 mCi) was collected. The product was concentrated to dryness under reduced pressure, and the radiolabeled product injection solution was prepared by adding physiological saline (containing 2.5% (v / v) Tween 80 and 2.5% (v / v) ascorbic acid). The radiochemical purity was no less than 95%.

[0135] Labeling Synthesis Examples 2-4:

[0136] 131 I-labeled (3-iodo-4-methoxyphenyl)(1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone ( 131 I-KIDJI-008) labeled synthesis

[0137] The product was prepared according to the method of labeled synthesis example 2-3 using (4-methoxy-3-(tri-n-butyltin)phenyl)(1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone as the labeled precursor, with a radiochemical purity of >95%.

[0138] Labeling Synthesis Example 2-5:

[0139] 131 I-labeled (2-iodo-4-nitro)(1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone ( 131 I-KIDJI-009) labeled synthesis

[0140] Prepared according to the method of Labeling Synthesis Example 2-3, using (4-nitro-2-(tri-n-butyltinyl)phenyl)(1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone as the labeled precursor. Radiochemical purity >95%.

[0141] Labeling Synthesis Example 2-6:

[0142] 131 I-labeled (2-fluoro-4-iodophenyl)(1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone ( 131 The labeled synthesis of I-KIDJI-010) was prepared according to the method of labeled synthesis example 2-3 using (2-fluoro-4-(tri-n-butyltin)phenyl)(1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone as the labeled precursor, with a radiochemical purity of >95%.

[0143] Example 3: 18 Ex vivo autoradiography of F-KIDJI-003

[0144] The spontaneous type 2 diabetes mouse model was constructed using db / db homozygous mice (BKS.Cg-Dock7m+ / +LeprdbJ; Mice Strain Code 607. A defect in the Leptin receptor gene on chromosome 4 causes db / db mice to exhibit hyperphagia and obesity from four weeks of age. Over time, these mice develop significant hyperglycemia, hyperlipidemia, and insulin resistance, a pathogenesis similar to that seen in subjects with type 2 diabetes. Control mice were either normal (m / m) or heterozygous (m / db) mice.

[0145] Mice were anesthetized with 1.5% (v / v) isoflurane and injected into the tail vein 18 F-KIDJI-003 (15 MBq). After 60 minutes, mice were killed by dislocation, and the kidneys were removed and quickly frozen in dry ice. 20 μm frozen sections were made using a freezing microtome (RWD Minux FS800). The frozen sections were sealed with a developing plate (FUJIFILM; BAS-IP SR 2025E) for 2 hours, and images were read and quantitatively analyzed using a radioautography scanner (CR-35 Bio; Elysia-Raytest).

[0146] Figure 1 is an ex vivo autoradiography. Compared with normal mice, 18 F-KIDJI-003 showed significant accumulation in the renal pelvis of diabetic nephropathy mice. The results of quantitative analysis (Figure 2) showed that F-KIDJI-003 showed significant accumulation in the renal pelvis of diabetic nephropathy mice. 18The radioactivity of F-KIDJI-003 was more than four times that of the renal pelvis of normal mice. Data were collected from the left and right kidneys of two normal mice and two diabetic nephropathy mice, with a total of four kidneys in each group.

[0147] Example 4: In vivo imaging

[0148] In vivo imaging example 4-1: 18 In vivo PET imaging of F-KIDJI-004

[0149] PET scanning was performed using an Inveon scanner (Siemens Medical Solutions Knoxville, TN, USA). Mice (m / m and db / db mice) were anesthetized with 1.5% (v / v) isoflurane and fixed in the center of the FOV of the PET scanner. 18 Immediately after administration of F-KIDJI-004 (15 MBq), radiofrequency signals were acquired in 3D mode for 90 minutes. The energy window was 350-750 keV. Following the PET scan, mice were intravenously injected with 5 μl / g body weight of ioversesol (35%) (Optiray 350; Guerbet), followed by a 30-second CT scan (X-ray source: 70 kV / 88 mA, FOV: 60 mm). During the PET and CT scans, mice were anesthetized with 1.5% (v / v) isoflurane.

[0150] In vivo imaging example 4-2 131 SPECT in vivo imaging of I-KIDJI-007

[0151] SPECT scanning was performed using a nanoScan SPECT / CT scanner (Mediso Medical Imaging Systems, Hungary). Mice were anesthetized with 1.5%-2.0% (v / v) isoflurane and placed on a preheated scanning bed. 131 I-KIDJI-007 (11.1 MBq) was administered, and then radioactivity was acquired in 3D mode every 30 minutes for 2 hours. The energy window was set to 360-700 keV. After the SPECT scan was completed, 5 μL / g body weight of ioversol injection (Optiray 350; Guerbet) was injected through the tail vein of the mouse, followed by a 30-second CT scan (X-ray source: 50 kV / 980 μA, 480 projections; mice were anesthetized with 1.5% (v / v) isoflurane during the SPECT and CT scans.

[0152] In vivo imaging example 4-3131 In vivo imaging of I-KIDJI-008

[0153] 131 The in vivo imaging of I-KIDJI-008 was performed in a similar manner to that of Example 4-2. 131 I-KIDJI-007 is replaced by 131 I-KIDJI-008 was used for imaging.

[0154] In vivo imaging example 4-4

[0155] 131 In vivo imaging of I-KIDJI-009

[0156] 131 The in vivo imaging of I-KIDJI-009 was performed in a similar manner to that of Example 4-2. 131 I-KIDJI-007 is replaced by 131 I-KIDJI-009 was used for imaging.

[0157] In vivo imaging examples 4-5

[0158] 131 In vivo imaging of I-KIDJI-010

[0159] 131 The in vivo imaging of I-KIDJI-010 was performed in a similar manner to that of Example 4-2. 131 I-KIDJI-007 is replaced by 131 I-KIDJI-010 was used for imaging.

[0160] The results of in vivo imaging Example 4 are shown in Figure 3. The imaging results show that 18 F-KIDJI-004, 131 I-KIDJI-007、 131 I-KIDJI-008、 131 I-KIDJI-009 and 131 I-KIDJI-010 was significantly accumulated in the renal pelvis of the kidneys of db / db model mice, but not in the kidneys of normal mice.

[0161] Example 5: In vitro autoradiography of mouse kidney tissue sections

[0162] Fresh frozen kidneys of 6-month-old normal mice (m / m), diabetic nephropathy mice heterozygous (m / db) and homozygous (db / db) were prepared into frozen sections with a thickness of 20 μm. The obtained labeled protein obtained in Example 2-1 was added to 50 mM Tris-HCl buffer (pH 7.4). 18 F-KIDJI-003 was used as an incubation solution (final chemical concentration 5 nM). Kidney sections were immersed in incubation solution containing or not containing non-radiolabeled KIDJI-003 (10 μM) and allowed to stand for 1 hour. The sections were then washed twice for 2 minutes in 50 mM Tris-HCl buffer (pH 7.4). The sections were sealed in a development plate for 1 hour, and images were read and quantitatively analyzed using an autoradiography scanner (CR-35 Bio; Elysia-Raytest).

[0163] Table 1

[0164] The autoradiogram in the upper part of Figure 4 shows 18 F-Total binding (TB) of KIDJI-003 on fresh-frozen kidney sections of normal mice and diabetic nephropathy mouse models. The lower panel is an in vitro autoradiography with the addition of non-radiolabeled KIDJI-003, showing 18 Non-specific binding (NSB) of F-KIDJI-003 on fresh frozen kidney sections of normal mice and diabetic nephropathy mice. Specific binding (SB) is calculated by subtracting NSB from TB, which is a parameter that reflects P2X7R expression. Table 1 shows the quantitative results of Figure 4. The addition of non-radiolabeled KIDJI-003 significantly reduced 18 F-KIDJI-003 total binding showed a large number of 18 F-KIDJI-003 specifically binds. However, there is no difference in specific binding per unit area in the kidneys of normal mice and diabetic nephropathy mice. This indicates that P2X7R expression is not increased in diabetic nephropathy mice. Therefore, 18 The accumulation of F-KIDJI-003 in the renal pelvis of diabetic nephropathy mouse models is not due to 18 Combination of F-KIDJI003 and P2X7R.

[0165] Example 6 Immunostaining experiment of mouse kidney tissue sections

[0166] Six-month-old normal mice (m / m), heterozygous (m / db) and homozygous (db / db) mice were killed by cervical dislocation. The left and right kidneys were removed and immersed in 4% PFA / PBS fixative overnight to complete tissue fixation. After PBS washing, the tissues were immersed in 20% sucrose / PBS solution overnight and 30% sucrose / PBS solution overnight to complete freezing protection. The kidneys were then made into 10 μm thick frozen sections using a freezing microtome (Rayward). P2X7R staining was performed using anti-P2X7R (Cat#: APR-004, Alomone labs) antibody. After the kidney sections were treated with an autoclave (citric acid buffer (0.01M sodium citrate: 0.01M citric acid = 5:1), 121°C, for 5 minutes), they were washed with running water for 5 minutes, and TSA blocking buffer (TSA Fluorescein System, NEL70000, Perkin Elmer) was added to the kidney sections, and the sections were allowed to stand for 1 hour. An incubation solution containing the primary antibody (anti-P2X7R antibody, 1:1000) was added and allowed to stand overnight. The primary antibody was discarded, and the washing was repeated three times for 5 minutes with PBS. An incubation solution containing a biotin-labeled secondary antibody was added, and the sections were allowed to stand for another 1 hour. Thereafter, the fluorescent signal was amplified using a TSA sensitization kit (TSA Fluorescein System, NEL70000; Perkin Elmer), and the sections were blocked with VECTASHILD mounting medium (H-1000, Vector Laboratories Inc.) and observed under a microscope. The results are shown in Figure 5. There was no significant difference in P2X7R expression in the kidneys of 6-month-old normal mice (left), diabetic nephropathy mice heterozygous db / m (middle) and homozygous db / db (right). This result is consistent with the experimental results in Figure 4, once again showing that P2X7R expression is not increased in diabetic nephropathy model db / db homozygous mice. Therefore, it once again proves that 18 The accumulation of F-KIDJI003 in the renal pelvis of diabetic nephropathy mouse models is not due to 18 Combination of F-KIDJI003 and P2X7R.

[0167] Example 7 Diagnostic Method for Diabetic Nephropathy

[0168] In the diagnosis of DKD in subjects, the drug was injected intravenously as in animal studies. 18 After the F-KIDJI003 radioactive molecular probe is injected, PET or SPECT imaging is performed to diagnose diabetic nephropathy based on whether the radioactive molecular probe accumulates in the renal pelvis.

[0169] While the present invention has been illustrated and described with reference to certain preferred embodiments thereof, those skilled in the art will appreciate that the foregoing description is provided as a further detailed description of the present invention in conjunction with specific embodiments thereof, and that the present invention is not limited to these descriptions. Within the spirit and scope of the present invention, those skilled in the art may make various changes in form and detail, including simple deductions or substitutions, which also constitute a part of the present invention.

Claims

1. A compound represented by general formula (I) or a pharmaceutically acceptable salt, precursor or solvate thereof, in, X is carbon or nitrogen; R 1 is selected from hydrogen or alkyl; R 2 is one or more substituents on the benzene ring or pyridine ring, R 2 is independently selected from one or more of hydrogen, halogen, hydroxyl, cyano, nitro, amino, C1-C6 alkyl, C3-C6 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C6 cycloalkyloxy, C1-C6 alkylamino, C3-C6 cycloalkylamino, C1-C6 hydroxyalkyl, C3-C6 hydroxycycloalkyl, halogenated C1-C6 alkyl, halogenated C3-C6 cycloalkyl, halogenated C1-C6 alkoxy, halogenated C3-C6 cycloalkyloxy, halogenated C1-C6 alkylamino, halogenated C3-C6 cycloalkylamino, C6-C8 aryl or C5-C8 heteroaryl, and R 2 At least one atom of these substituents represented is substituted by a radionuclide or the entire substituent is substituted by a radionuclide.

2. The compound according to claim 1 or a pharmaceutically acceptable salt, precursor or solvate thereof, wherein in the general formula (I), R 1 Selected from hydrogen, C1-C6 alkyl or C3-C6 cycloalkyl.

3. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt, precursor, or solvate thereof, wherein the radionuclide is selected from 18 F. 11 C. 131 I. 123 I. 124 I. 125 One or more of I.

4. The compound according to claim 1 or a pharmaceutically acceptable salt, precursor, or solvate thereof, wherein the compound is selected from: (S)-(3-fluoro-2-trifluoromethylpyridin-4-yl)(6-methyl-1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone, (S)-(2-fluoro-6-methylpyridin-4-yl)(6-methyl-1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone, (4-iodophenyl)(1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone, (3-iodo-4-methoxyphenyl)(1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone, (2-iodo-4-nitro)(1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone, (2-Fluoro-4-iodophenyl)(1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone.

5. The compound according to claim 1 or a pharmaceutically acceptable salt, precursor, or solvate thereof, wherein the precursor compound is selected from: (S)-(3-chloro-2-trifluoromethylpyridin-4-yl)(6-methyl-1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone, (S)-(2-chloro-6-methylpyridin-4-yl)(6-methyl-1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone, (1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)(4-tri-n-butyltinphenyl)methanone, (4-methoxy-3-(tri-n-butyltinyl)phenyl)(1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone, (4-nitro-2-(tri-n-butyltinyl)phenyl)(1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone, (2-Fluoro-4-(tri-n-butyltinyl)phenyl)(1-(pyrimidin-2-yl)-1,4,6,7-tetrahydro-5H-[1,2,3]triazolo[4,5-c]pyridin-5-yl)methanone. 6 . The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the pharmaceutically acceptable salt is an addition salt of an inorganic acid or an organic acid.

7. The compound according to claim 6 or a pharmaceutically acceptable salt thereof, wherein the inorganic acid is selected from hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid and nitric acid, and the organic acid is selected from acetic acid, tartaric acid, salicylic acid, methanesulfonic acid, succinic acid, lemon segment, malic acid, lactic acid and fumaric acid.

8. The solvate of the compound according to claim 1, which is a hydrate.

9. Use of the compound according to any one of claims 1 to 8 or a pharmaceutically acceptable salt, precursor or solvate thereof in the preparation of a diagnostic drug for diabetic nephropathy.

10. A radioactive diagnostic kit for diagnosing diabetic nephropathy, the radioactive diagnostic kit comprising a compound having a structure represented by general formula (I) or a pharmaceutically acceptable salt, precursor or solvate thereof, in, X is selected from carbon (C) or nitrogen (N); R 1 is selected from hydrogen or alkyl; R 2 is one or more substituents on the benzene ring or pyridine ring, R 2 is independently selected from one or more of hydrogen, halogen, hydroxyl, cyano, nitro, amino, C1-C6 alkyl, C3-C6 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 alkoxy, C3-C6 cycloalkyloxy, C1-C6 alkylamino, C3-C6 cycloalkylamino, C1-C6 hydroxyalkyl, C3-C6 hydroxycycloalkyl, halogenated C1-C6 alkyl, halogenated C3-C6 cycloalkyl, halogenated C1-C6 alkoxy, halogenated C3-C6 cycloalkyloxy, halogenated C1-C6 alkylamino, halogenated C3-C6 cycloalkylamino, C6-C8 aryl or C5-C8 heteroaryl, and R 2 At least one atom of these substituents represented is substituted by a radionuclide or the entire substituent is substituted by a radionuclide.

11. The radioactive diagnostic kit according to claim 9, wherein the pharmaceutically acceptable salt is an addition salt of an inorganic acid or an organic acid, and the solvate is a hydrate.

12. The radioactive diagnostic kit according to claim 9, wherein the compound is selected from:

13. A method for diagnosing diabetic nephropathy, comprising: The compound of any one of claims 1 to 4, 6 to 8, or a pharmaceutically acceptable salt or solvate thereof is formulated into a physiologically acceptable solution, or the precursor of claim 5 is formulated into a physiologically acceptable solution after being labeled with a radionuclide; introducing the solution into a subject; Imaging the subject's kidneys; An analysis is performed based on the imaging results, and a diagnosis is made based on the analysis results.

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