Nitrogen-containing compound as well as preparation method and application thereof

Abstract

The invention discloses a nitrogen-containing compound as well as a preparation method and application thereof. The invention specifically discloses a compound as shown in a formula I or pharmaceutically acceptable salt thereof, which has one or more advantages as follows: the compound is good in targeting and can be used for developing diagnosis and / or treatment of prostatic cancer. # imgabs0 #

Description

Technical Field

[0001] The invention relates to a nitrogen-containing compound and a preparation method and application thereof. Background Art

[0002] Prostate cancer is the second most common type of cancer in men. Prostate-specific membrane antigen-positive, metastatic castration-resistant prostate cancer (PSMA-positive mCRPC) is the terminal stage of prostate cancer disease development, with an insufficient response to castration therapy, leading to prostate cancer recurrence and metastasis. Currently, there are few treatment options for this type of patient, and the 5-year survival rate is low. Prostate-specific membrane antigen (PSMA) is highly expressed in more than 80% of prostate cancer patients. Combining targeted compounds (ligands) with diagnostic or therapeutic radionuclides allows the drug to bind to prostate cancer cells expressing PSMA. The gamma or beta rays emitted when the radioactive diagnostic or therapeutic nuclides decay can image and treat diseased tissues.

[0003] In view of the importance of prostate cancer diagnosis and treatment, there is an urgent need to develop a radioactive compound with good targeting that can be used for prostate cancer imaging diagnosis and / or treatment. Summary of the invention

[0004] The technical problem to be solved by the present invention is to overcome the defect of the single compound for diagnosing and treating prostate cancer in the prior art. To this end, the present invention provides a nitrogen-containing compound and a preparation method and use thereof. The compound of the present invention has one or more of the following advantages: good targeting and can be used for imaging diagnosis and / or treatment of prostate cancer.

[0005] The present invention provides a compound I or a pharmaceutically acceptable salt thereof,

[0006]

[0007] Among them, A is composed of a chelating group and a radionuclide;

[0008] X is (*Position directly connected to L 2 connected);

[0009] n1 is an integer selected from 1 to 20;

[0010] n2 is an integer selected from 1 to 10;

[0011] L 1 and L 2 Independently -C 1 -C 6 Alkylene-;

[0012] R is

[0013] In a certain embodiment of the present invention, the compound I is

[0014] Preferably, the compound I is selected from the following structures:

[0015] More preferably, the compound I is selected from the following structures:

[0016] In a certain embodiment of the present invention, in A, the chelating group is a conventional chelating group in the art; preferably, the chelating group is 1,4,7,10-tetraazacyclododecane-N,N',N",N"'-tetraacetic acid (DOTA), 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA), 2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecane-1-yl)pentanedioic acid (DOTA-GA ), 2-(4,7-bis(carboxymethyl)-1,4,7-triazonon-1-yl) glutaric acid (NODA-GA), diethylenetriaminepentaacetic acid (DTPA), N,N′-bis-[2-hydroxy-5-(carboxyethyl)benzyl]ethylenediamine-N,N′-diacetic acid (HBED-CC) or thioacetyltriglycine (MAG3) structure, the structure formed by removing a hydroxyl group from a carboxyl group, for example, the structure formed by removing a hydroxyl group from a carboxyl group of DOTA

[0017] In one embodiment of the present invention, n1 is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16, for example, n1 is an integer from 1 to 10, and for example, n1 is 2.

[0018] In one embodiment of the present invention, n2 is 1, 2, 3, 4, 5, 6, 7, 8 or 9, for example, n2 is an integer from 1 to 5, and for example, n2 is 1.

[0019] In one embodiment of the present invention, L 1 and L 2 In the “-C 1 -C 6 Alkylene-" is independently -C 1 -C 3 Alkylene-, e.g.

[0020] In one embodiment of the present invention, L 1 In the “-C 1 -C 6 Alkylene-"

[0021] In one embodiment of the present invention, L 2 In the “-C 1 -C 6 Alkylene-"

[0022] In one embodiment of the present invention, in A, the chelating group chelates with a radionuclide.

[0023] In a certain embodiment of the present invention, the radionuclide is a diagnostic nuclide or a therapeutic nuclide.

[0024] In a certain embodiment of the present invention, the diagnostic nuclide is 18 F. 68 Ga, 99m Tc or 64 Cu.

[0025] In a certain embodiment of the present invention, the therapeutic nuclide is 177 Lu, 188 Re, 225 Ac, 212 Pb, 211 At or 67 Cu.

[0026] In a certain embodiment of the present invention, the radionuclide is 18 F. 68 Ga, 177 Lu, 99m Tc, 188 Re, 64 Cu, 67 Cu, 225 Ac, 212 Pb or 211 At.

[0027] In a certain embodiment of the present invention, the valence state of the radionuclide is monovalent, divalent, trivalent or tetravalent, for example, trivalent.

[0028] In a certain embodiment of the present invention, A is composed of and a radionuclide, wherein the radionuclide is 177 Lu or 68 Ga; preferably, A is composed of and a radionuclide chelate, wherein the radionuclide is 177 Lu or 68 Ga, e.g.

[0029] In a certain embodiment of the present invention, the structure of compound I is

[0030]

[0031] The present invention also provides a compound II or a pharmaceutically acceptable salt thereof,

[0032]

[0033] Among them, B is composed of a chelating group and a non-radioactive nuclide;

[0034] The chelating group, X, L 1 , L 2 and R are defined as described in any of the previous schemes.

[0035] In a certain embodiment of the present invention, the non-radioactive nuclide is F, Ga, Lu, Tc, Re, Cu, Cu, Ac, Pb or At.

[0036] In a certain embodiment of the present invention, the compound II is

[0037]

[0038] The present invention also provides a compound III or a pharmaceutically acceptable salt thereof,

[0039]

[0040] Wherein, C is a chelating group;

[0041] The chelating group, X, L 1 , L 2 and R are defined as in any of the previous schemes.

[0042] In a certain embodiment of the present invention, the compound III is

[0043] The present invention also provides a pharmaceutical composition, which comprises a substance D and a pharmaceutical excipient, wherein the substance D is the compound I, compound II, compound III or a pharmaceutically acceptable salt thereof ("it" represents the compound I, compound II or compound III).

[0044] The present invention also provides a kit, which comprises a substance D and instructions, wherein the substance D is the compound I, compound II, compound III or a pharmaceutically acceptable salt thereof ("it" represents the compound I, compound II or compound III).

[0045] The present invention also provides a use of a substance D in the preparation of a medicament for treating a PSMA-related disease, wherein the substance D is the compound I, compound II, compound III or a pharmaceutically acceptable salt thereof ("it" represents the compound I, compound II or compound III); the PSMA-related disease is preferably prostate cancer, more preferably prostate cancer with high PSMA expression or PSMA-positive mCRPC (prostate-specific membrane antigen-positive, metastatic castration-resistant prostate cancer).

[0046] The present invention also provides a use of the compound I, compound III or a pharmaceutically acceptable salt thereof ("it" represents the compound I or compound III) in the preparation of a drug for treating and / or preventing cancer.

[0047] The cancer is preferably prostate cancer, more preferably prostate cancer with high PSMA expression or PSMA-positive mCRPC (prostate-specific membrane antigen-positive, metastatic castration-resistant prostate cancer).

[0048] The present invention also provides a use of the compound I, compound III or a pharmaceutically acceptable salt thereof ("it" represents the compound I or compound III) in the preparation of an imaging agent.

[0049] The imaging agent is preferably an imaging agent for diagnosing cancer; the cancer is preferably prostate cancer.

[0050] The present invention also provides a compound IV or a pharmaceutically acceptable salt thereof,

[0051]

[0052] Among them, L 1 The definition of is as described in any of the previous schemes.

[0053] In a certain embodiment of the present invention, the compound IV is

[0054]

[0055] Terminology explanation:

[0056] The term "pharmaceutically acceptable salt" refers to a salt obtained by reacting a compound with a pharmaceutically acceptable (relatively non-toxic, safe, and suitable for use by patients) acid or base. When the compound contains a relatively acidic functional group, a base addition salt can be obtained by contacting the free form of the compound with a sufficient amount of a pharmaceutically acceptable base in a suitable inert solvent. When the compound contains a relatively basic functional group, an acid addition salt can be obtained by contacting the free form of the compound with a sufficient amount of a pharmaceutically acceptable acid in a suitable inert solvent.

[0057] The term "alkyl" refers to a group having a specified number of carbon atoms (e.g., C 1 ~C 6 ) is a straight or branched chain alkyl group. The alkyl group includes, but is not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, etc.

[0058] In the present invention, the term "alkylene" refers to a saturated straight-chain or branched divalent hydrocarbon group. 1-6 Alkylene refers to an alkylene group having 1 to 6 carbon atoms, and specific examples thereof include methylene, ethylene (e.g., -CH 2 CH 2 -、-CH(CH 3 )-), propylene (e.g. -CH 2 CH 2 CH 2 -、-C(CH 3 ) 2 -、-CH 2 CH(CH 3 )-), butylene (e.g. -CH 2 CH 2 CH 2 CH 2 -、-CH(CH 3 )CH(CH 3 )-、-CH 2 CH(CH 3 )CH 2 -), n-pentylidene or n-hexylidene.

[0059] In the present invention, the term "alkyleneoxy" refers to -O-alkylene-, wherein alkylene is as defined above.

[0060] In the present invention, the term "alkylenethio" refers to -S-alkylene-, wherein alkylene is as defined above.

[0061] The term "aryl" refers to a radical having a specified number of carbon atoms (e.g. 6 ~C 10 ) and a cyclic group consisting only of carbon atoms, which is a monocyclic or condensed ring. Aryl includes but is not limited to phenyl or naphthyl.

[0062] The terms "pharmaceutically acceptable excipients" and "pharmaceutical excipients" refer to excipients and additives used in the production of drugs and the preparation of prescriptions. They are all substances contained in drug preparations except active ingredients. For details, please refer to the Pharmacopoeia of the People's Republic of China (2020 edition) or Handbook of Pharmaceutical Excipients (Raymond C Rowe, 2009).

[0063] The term "treat" refers to any of the following: (1) alleviating one or more biological manifestations of a disease; (2) interfering with one or more points in the biological cascade that initiates a disease; or (3) slowing the progression of one or more biological manifestations of a disease.

[0064] Without violating the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0065] The reagents and raw materials used in the present invention are commercially available.

[0066] The positive and progressive effects of the present invention are that the compounds of the present invention have one or more of the following advantages: good targeting and can be used for imaging diagnosis and / or treatment of prostate cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0067] Figure 1 for 177 SPECT / CT images of Lu-DOTA-Tri-PSMA at different time points in mice, corresponding to 0.5h, 1h, 2h, 4h, 6h, 24h, 48h, 72h, 96h and 120h, respectively. Figure 1 a, b, c, d, e, f, g, h, i and j parts. DETAILED DESCRIPTION

[0068] The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples. The experimental methods in the following examples without specifying specific conditions are carried out according to conventional methods and conditions, or selected according to the product specifications.

[0069] Example 1 Radionuclides 177 Lu Marking DOTA-N 3

[0070] DOTA-N 3 Solution preparation: DOTA-N 3 (Formula 1) was dissolved in 0.5M acetic acid-sodium acetate buffer solution with a pH of 5.2±0.1, wherein DOTA-N 3 The concentration is 0.2 mg / mL.

[0071] Add 26 μL 177 LuCl 3 Solution (radioactivity of about 18 mCi, solvent 0.04 mol / L hydrochloric acid) and 150 μL DOTA-N 3 The solutions were mixed and heated at 95°C for 15 min to obtain 177 Lu-DOTA-N 3The reaction solution was 176 μL, and the labeling rate was 87.82% by Radio-HPLC, as shown in Table 1 below; wherein the chromatographic peak 4 is 177 Lu-DOTA-N 3 The corresponding chromatographic peaks.

[0072]

[0073] Radio-HPLC detection conditions:

[0074] Chromatographic column: ZORBAX Eclipse Plus C18 (4.6 mm × 250 mm, 5 μm) Mobile phase: A: 0.1% TFAinH 2 O, B: 0.1% TFA in CH 3 CN

[0075] Gradient: 0-5-10-14-15-17 min, 1-1-10-10-1-1% B

[0076] Flow rate: 1mL / min

[0077] Wavelength: 220nm

[0078] Column temperature: 30°C

[0079] Table 1

[0080]

[0081] Example 2 Click Chemistry Reaction

[0082] Synthesis route of PSMA-alkyne:

[0083]

[0084] Compd1 (1 eq) was dissolved in DMF, and then DIPEA (5 eq) and Compd2 (2 eq) were added in sequence. The mixture was fully reacted at room temperature for 2 hours and monitored by LC-MS. After the reaction of the raw materials was complete, the mixture was concentrated in vacuo, dried, and purified by reverse phase chromatography to obtain the target product PSMA-alkynyl.

[0085] Preparation of PSMA-alkynyl solution: The above PSMA-alkynyl was dissolved in DMF, wherein the concentration of PSMA-alkynyl was 0.7 mg / mL.

[0086] Preparation of copper sulfate solution: copper sulfate was dissolved in 0.5 M acetic acid-sodium acetate buffer solution with a pH of 5.2±0.1, wherein the concentration of copper sulfate was 3.2 mg / mL.

[0087] Preparation of sodium ascorbate solution: Sodium ascorbate was dissolved in 0.5 M acetic acid-sodium acetate buffer solution with a pH of 5.2±0.1, wherein the concentration of sodium ascorbate was 17.5 mg / mL.

[0088] 95 μL of PSMA-alkyne solution was mixed with the 177 Lu-DOTA-N 3 The reaction solution 175 μL, 43 μL copper sulfate solution, 45 μL sodium ascorbate solution were mixed and heated at 40°C for 60 min to obtain 177 358 μL of Lu-DOTA-Tri-PSMA reaction solution was detected by Radio-HPLC under the same analytical conditions. 177 The peak position of Lu-DOTA-Tri-PSMA and the cold reference compound 175 Lu-DOTA-Tri-PSMA (cold reference compound uses non-radioactive raw materials and uses 177 The product was prepared by a similar method to Lu-DOTA-Tri-PSMA; the molecular weight and 175 The theoretical value of Lu-DOTA-Tri-PSMA is consistent; confirmed by Radio-HPLC 175 The peak position of Lu-DOTA-Tri-PSMA) is consistent with the peak position of Lu-DOTA-Tri-PSMA. The labeling rate detected by Radio-HPLC is 72.79%, see Table 2 below; 177 Chromatographic peak corresponding to Lu-DOTA-Tri-PSMA.

[0089] The aforementioned 177 358 μL of the Lu-DOTA-Tri-PSMA reaction solution was diluted to 2 mL with sterile water for injection to obtain the solution to be purified. 2 mL of the solution to be purified was purified by C18 column to obtain the target product with a radiochemical purity of 92.78% ( 177 Lu-DOTA-Tri-PSMA ethanol solution), see Table 3 below, where chromatographic peak 5 is 177 Chromatographic peak corresponding to Lu-DOTA-Tri-PSMA.

[0090]

[0091] Radio-HPLC detection conditions:

[0092] Chromatographic column: ZORBAX Eclipse Plus C18 (4.6mm×250mm, 5μm)

[0093] Mobile phase: A: 0.1% TFA in H 2 O, B: 0.1% TFA in CH3 CN

[0094] Gradient: 0-10-20-30-35-38-40-45 min, 1-10-20-40-40-100-1-1% B

[0095] Flow rate: 1mL / min

[0096] Wavelength: 220nm

[0097] Column temperature: 30°C

[0098] C18 column purification conditions (C18 column model: Waters, Sep-Pak Light-C18 solid phase extraction column):

[0099] Activation of C18 column: First, rinse the C18 column with 5 mL of ethanol, and then rinse with 5 mL of sterile water for injection.

[0100] Purify the product with C18 column: pass 2 mL of the above-mentioned solution to be purified through C18 column (i.e., elute it in C18 column); elute C18 column with 1.5 mL of sterile water for injection to remove radioactive impurities; elute C18 column with 0.5 mL of anhydrous ethanol, collect the eluate with higher concentration, and obtain the target product: 177 0.4 mL of Lu-DOTA-Tri-PSMA ethanol solution.

[0101] Table 2

[0102]

[0103]

[0104] Table 3

[0105]

[0106] Example 3 In vitro stability

[0107] 60 μL of the target product obtained in Example 2 was diluted with 1 mL of 0.5 M acetic acid-sodium acetate buffer solution with a pH of 5.2 and placed in a 25° C. stability test chamber. The initial radiochemical purity of the target product was 92.78%. After 24 hours, the radiochemical purity of the target product remained essentially unchanged at 92.07%.

[0108] Example 4 Biodistribution

[0109] Preparation of radioactive injection solution 1: The target product obtained in Example 2 (i.e. 177 The ethanol solution of Lu-DOTA-Tri-PSMA) was diluted with 0.5 M acetic acid-sodium acetate buffer at pH 5.2 to a radioactivity concentration of about 1 mCi / mL.

[0110] 22Rv1 male tumor-bearing nude mice (Shanghai Junna Medical Technology Co., Ltd., catalog number: NO.202373380) were used, and the aforementioned radioactive injection solution 1 was injected into the mice through the tail vein (about 100 μL, 100 μCi / mouse, 4 mice / group, four groups in total), and the animals were killed at 0.5 h, 1 h, 4 h and 24 h after injection, respectively. The tissues and organs of interest were dissected and weighed, and the radioactive counts were measured using a gamma counter and the ID% / g of the tissues and organs were calculated (calculation formula: ID% / g=tissue count / total count in the injected mouse / tissue weight*100%).

[0111] In this embodiment, 24 hours after administration, 177 The organ distribution of Lu is shown in Table 4 below, which is consistent with the compounds reported in the literature. 177 Compared with Lu-PSMA-617 (Wu, Y.; Zhang, X.; Duan, X.; Yang, X.; Wang, F.; Zhang, J. OptimizedTherapeutic 177 Lu-Labeled PSMA-Targeted Ligands with Improved PharmacokineticCharacteristics for Prostate Cancer.Pharmaceuticals2022,15,1530), 177 Lu-DOTA-Tri-PSMA showed high PSMA-specific tumor uptake: kidney uptake was close to complete metabolism after 24 h (1.58±0.70ID% / g), while tumor uptake remained high (9.14±3.16ID% / g); other organs, such as liver (0.07±0.02ID% / g), spleen (0.05±0.02ID% / g) and lung (0.06±0.01ID% / g), showed very low uptake. 177 The favorable pharmacokinetics of Lu-DOTA-Tri-PSMA resulted in a high tumor-to-background ratio after 24 h (tumor / blood: 304.67; tumor / muscle: 914.00).

[0112] At different time points after drug administration, 177 The comparison of Lu uptake in various tissues of 22Rv1 tumor-bearing mice is shown in Table 5. It can be seen that 177 Lu is rapidly cleared from all major organs and is specifically taken up by tumor tissue, so the tumor-to-background ratio increases over time.

[0113]

[0114] Table 4 177 Lu-DOTA-Tri-PSMA 177 Comparison of Lu-PSMA-617 uptake in various tissues of 22Rv1 tumor-bearing mice 24 hours after administration

[0115] <![CDATA[ 177 Lu-DOTA-Tri-PSMA(ID% / g)]]> <![CDATA[ 177 Lu-PSMA-617(ID% / g)]]> Blood 0.03±0.00 0.24±0.11 Heart 0.02±0.00 0.10±0.03 liver 0.07±0.02 0.56±0.14 spleen 0.05±0.02 0.78±0.22 lung 0.06±0.01 0.33±0.10 kidney 1.58±0.70 2.67±1.02 Salivary glands 0.04±0.01 0.19±0.05 muscle 0.01±0.00 0.08±0.01 bone 0.03±0.02 0.40±0.13 Tumor 9.14±3.16 3.74±0.29 Tumor / blood 304.67 15.58 Tumor / Muscle 914.00 46.75 Tumor / kidney 5.78 1.40

[0116] Table 5 177 Comparison of Lu-DOTA-Tri-PSMA uptake in various tissues of 22Rvl tumor-bearing mice at different time points after administration (unit: ID% / g)

[0117] 0.5h 1h 4h 24h Blood 3.27±0.09 1.08±0.25 0.16±0.04 0.03±0.00 Heart 1.03±0.32 0.39±0.08 0.06±0.01 0.02±0.00 liver 1.46±0.25 0.74±0.07 0.18±0.06 0.07±0.02 spleen 3.61±0.66 2.11±0.22 0.24±0.14 0.05±0.02 lung 2.98±0.90 1.16±0.28 0.18±0.04 0.06±0.01 kidney 99.49±26.18 64.50±5.38 8.56±2.55 1.58±0.70 Salivary glands 1.62±0.44 0.65±0.15 0.15±0.03 0.04±0.01 muscle 0.91±0.22 0.28±0.10 0.08±0.05 0.01±0.00 bone 0.76±0.34 0.30±0.15 0.07±0.02 0.03±0.02 Tumor 19.75±3.02 19.00±0.65 14.20±1.45 9.14±3.16 Tumor / blood 6.04 17.59 88.75 304.67 Tumor / Muscle 21.70 67.86 177.50 914.00 Tumor / kidney 0.20 0.29 1.66 5.78 .

[0118] Example 5 Small Animal SPECT / CT Imaging

[0119] Preparation of radioactive injection solution 2: The target product obtained in Example 2 (i.e. 177 The ethanol solution of Lu-DOTA-Tri-PSMA) was diluted with 0.5 M acetic acid-sodium acetate buffer at pH 5.2 to a radioactivity concentration of about 4.75 mCi / mL.

[0120] 22Rv1 male tumor-bearing nude mice (Shanghai Junna Medical Technology Co., Ltd., Catalog No.: NO.202373380) were used to inject radioactive labeled compounds 177 Lu-DOTA-Tri-PSMA was injected into mice via the tail vein (about 120 μL, 600 μCi / mouse, 5 mice in total), and the animals were scanned in vivo at 0.5 h, 1 h, 2 h, 4 h, 6 h, 24 h, 48 h, 72 h, 96 h, and 120 h after injection.

[0121] 177 SPECT / CT imaging of Lu-DOTA-Tri-PSMA showed early accumulation in the bladder and kidney uptake, rapid background clearance, significant reduction in kidney accumulation after 4 hours, substantial clearance in the kidney and bladder after 6 hours, and complete clearance in the kidney and bladder after 24 hours. 177 Lu-DOTA-Tri-PSMA further accumulated and was retained in PSMA-expressing tumors, see Figure 1 ; The corresponding quantitative data are shown in Table 6 (unit: ID% / mm 3 ):

[0122] Table 6 In vivo scanning data of various organs in SPECT / CT imaging experiments

[0123] 0.5h 1h 2h 4h 6h 24h 48h 72h 96h 120h Tumor 3.51E-06 3.29E-06 3.73E-06 3.71E-06 2.87E-06 1.95E-06 3.08E-06 2.05E-06 9.88E-07 8.58E-07 kidney 3.19E-06 3.13E-06 1.59E-06 1.01E-06 6.12E-07 1.44E-07 3.78E-09 8.66E-09 1.12E-11 1.66E-09 bladder 2.85E-04 2.03E-04 1.17E-04 6.43E-06 1.87E-06 1.86E-08 2.35E-08 1.43E-08 9.27E-10 4.43E-15 .

[0124] Example 6 Treatment Effect

[0125] Take the obtained 177 Lu-DOTA-Tri-PSMA and 177 Lu-PSMA-617 drug was injected into two groups (4 mice / group) of tumor-bearing mice (22Rv1 male nude mice) through tail vein injection (injection volume: 100 μL / mouse, injection activity: 1.5 m Ci / mouse). The tumor size of mice was measured and the volume was calculated every other day together with the control group (4 mice, not injected with any drug, only injected with 0.9% NaCl aqueous solution). By comparing the tumor size, the therapeutic effect of the two drugs on mice was evaluated.

[0126] 177 Lu-DOTA-Tri-PSMA showed good therapeutic effects on cancer mice, especially prostate cancer mice.

[0127] The changes in tumor volume of mice in the experimental group and the control group over time are shown in Table 7 below:

[0128]

[0129]

[0130] The volume values ​​in the above table are the average tumor volumes of mice in each group, in mm 3 ;

[0131] “—” indicates that the first test mouse died at that time.

[0132] The changes in body weight of mice in the experimental group and the control group over time are shown in Table 8 below:

[0133]

[0134]

[0135] The weight values ​​in the above table are the average weights of mice in each group, in g;

[0136] “—” indicates that the first test mouse died at that time.

[0137] The survival of mice in the experimental group and the control group changes over time are shown in Table 9 below:

[0138]

[0139] Example 7 68 Preparation of Ga-DOTA-Tri-PSMA:

[0140] 1. Synthesis of DOTA-Tri-PSMA

[0141]

[0142] Compd (1 eq) was dissolved in DMF, and compd2 (1.2 eq) and DIEA (N, N-diisopropylethylamine) (3 eq) were added to the reaction solution. The reaction was carried out at room temperature for 1 hour, and the reaction was monitored by LC-MS. The raw material was reacted completely, and after being concentrated under reduced pressure, an appropriate amount of TFA was added for 5 minutes to remove the tBu protecting group, and the ether was washed twice, centrifuged, and dried to obtain the crude product compd3 (Yield: 32 mg, 70%)

[0143] Compd3 (1 eq) and compd4 (2 eq) were dissolved in EtOH / H 2 O(1:1),N 2 Under protection, CuSO 4 .5H 2 O (0.2eq) aqueous solution was added to the reaction solution, and finally sodium ascorbate (sodium ascorbate) (0.2eq) aqueous solution was added to the reaction solution. The reaction was carried out at room temperature for 2.5 hours, and the reaction was monitored by LC-MS. The raw material reacted completely, concentrated, and prepared by reverse phase to obtain compd5 (Yield: 28mg, 50%)

[0144] Compd5 (1 eq) was dissolved in DMF, and compd6 (1.5 eq) and DIEA (3 eq) were added to the reaction solution. The reaction was carried out at room temperature for 2 hours. The reaction was monitored by LC-MS. The raw material was reacted completely. The reaction was concentrated under reduced pressure and purified by reverse phase to obtain the target product DOTA-Tri-PSMA (Yield: 23 mg, 56%).

[0145] 2. Preparation of 0.5 mg / mL DOTA-Tri-PSMA solution

[0146] Take DOTA-Tri-PSMA, add 2000 μL of deionized water to dilute it, and mix well to obtain a precursor solution.

[0147] 3. 68 Preparation of Ga-labeled compounds

[0148] 1000 μL 68 GaCl 3 The solution (59.2MBq) was mixed with 150μL 1M NaOAc, and then 100μL of the aforementioned precursor solution (i.e., 0.5mg / mL DOTA-Tri-PSMA solution) was added thereto, and the mixture was thoroughly mixed. The mixture was incubated at 95°C for 15min. After the reaction was completed, the reaction bottle was cooled, and samples were taken for Radio-HPLC analysis.

[0149] 4. Radio-HPLC detection conditions

[0150] Chromatographic column: ZORBAX Eclipse Plus C18 (4.6 mm × 250 mm, 5 μm) Column temperature: 30 ° C Mobile phase: A: 0.1% TFA in H 2 O, B: 0.1% TFA in ACN

[0151] Gradient: 0-10-20-22-27-30-35 min, 10-20-40-40-100-10-10% B

[0152] Flow rate: 1mL / min

[0153] Wavelength: 220nm

[0154] 5. Radio-HPLC test results are shown in Table 10 below

[0155] Table 10

[0156]

[0157] 6. In vitro stability

[0158] The obtained target product was placed in a 25° C. stability test chamber. The initial radiochemical purity of the target product was 96.62%. After 4.5 hours, the radiochemical purity of the target product remained essentially unchanged at 96.89%.

Claims

1. A compound I or a pharmaceutically acceptable salt thereof, in, A is composed of a chelating group and a radionuclide; X is * is X and L 2 The connection location; n1 is an integer selected from 1 to 20; n2 is an integer selected from 1 to 10; L 1 and L 2 are independently -C1-C6 alkylene-; R is 2. The compound I or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: It meets one or more of the following conditions: (1) In A, the chelating group is a structure formed by removing a hydroxyl group from a carboxyl group based on the structure of DOTA, NOTA, DOTA-GA, NODA-GA, DTPA, HBED-CC or MAG3; (2) n1 is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16, for example, n1 is an integer from 1 to 10, and for example, n1 is 2; (3) n2 is 1, 2, 3, 4, 5, 6, 7, 8 or 9, for example, n2 is an integer from 1 to 5, and for example, n2 is 1; (4)L 1 and L 2 In the above, the "-C1-C6 alkylene-" is independently -C1-C3 alkylene-; (5) In A, the chelating group chelates with the radionuclide; (6) The radionuclide is a diagnostic nuclide or a therapeutic nuclide; (7) The radionuclide is 18 F. 68 Ga, 177 Lu, 99m Tc, 188 Re, 64 Cu, 67 Cu, 225 Ac, 212 Pb or 211 At; (8) The valence state of the radionuclide is monovalent, divalent, trivalent or tetravalent; (9) The compound I is 3. The compound I or a pharmaceutically acceptable salt thereof according to claim 2, characterized in that: It meets one or more of the following conditions: (1) In A, the chelating group is (2)L 1 In the above, the "-C1-C6 alkylene-" is (3)L 2 In the above, the "-C1-C6 alkylene-" is (4) The diagnostic nuclide is 18 F. 68 Ga, 99m Tc or 64 Cu; (5) The therapeutic nuclide is 177 Lu, 188 Re, 225 Ac, 212 Pb, 211 At or 67 Cu; (6) The valence state of the radionuclide is trivalent; (7) The compound I is selected from the following structures:

4. The compound I or a pharmaceutically acceptable salt thereof according to claim 3, characterized in that: It meets one or more of the following conditions: (1) A and a radionuclide, wherein the radionuclide is 177 Lu or 68 Ga; preferably, A is composed of and a radionuclide chelate, wherein the radionuclide is 177 Lu or 68 Ga, e.g. (2) The compound I is selected from the following structures:

5. The compound I or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The structure of compound I is 6. A compound II or a pharmaceutically acceptable salt thereof, in, B is composed of chelating groups and non-radioactive nuclides; The chelating group, X, L 1 , L 2 and R is defined as in any one of claims 1 to 4; Preferably, the non-radioactive nuclide is F, Ga, Lu, Tc, Re, Cu, Cu, Ac, Pb or At; Preferably, the compound II is 7. A compound III or a pharmaceutically acceptable salt thereof, in, C is a chelating group; The chelating group, X, L 1 , L 2 and R is defined as in any one of claims 1 to 4; Preferably, the compound III is 8. A pharmaceutical composition comprising a substance D and a pharmaceutical excipient, wherein the substance D is the compound I as described in any one of claims 1 to 5, the compound II as described in claim 6, the compound III as described in claim 7, or a pharmaceutically acceptable salt thereof.

9. A kit comprising a substance D and instructions, wherein the substance D is the compound I as described in any one of claims 1 to 5, the compound II as described in claim 6, the compound III as described in claim 7, or a pharmaceutically acceptable salt thereof.

10. Use of a substance D in the preparation of a drug for treating a PSMA-related disease, wherein the substance D is the compound I according to any one of claims 1 to 5, the compound II according to claim 6, the compound III according to claim 7, or a pharmaceutically acceptable salt thereof; The PSMA-related disease is preferably prostate cancer, more preferably prostate cancer with high PSMA expression or PSMA-positive mCRPC.

11. Use of the compound I according to any one of claims 1 to 5, the compound III according to claim 7 or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating and / or preventing cancer; The cancer is preferably prostate cancer, more preferably prostate cancer with high PSMA expression or PSMA-positive mCRPC.

12. Use of the compound I according to any one of claims 1 to 5, the compound III according to claim 7 or a pharmaceutically acceptable salt thereof in the preparation of an imaging agent; The imaging agent is preferably an imaging agent for diagnosing cancer; the cancer is preferably prostate cancer.

13. A compound IV or a pharmaceutically acceptable salt thereof, in, L 1 The definition as in any one of claims 1 to 4; The compound IV is preferably

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