PSMA targeting small molecule compound and preparation method and application thereof, nuclide labeling probe and preparation method and application thereof, pharmaceutical composition and application thereof, kit and application thereof

Abstract

The invention provides a PSMA targeting small molecule compound and a preparation method and application thereof, a nuclide labeling probe and a preparation method and application thereof, a pharmaceutical composition and application thereof, and a kit and application thereof, and relates to biological medicine. The PSMA targeted small molecule compound and the nuclide labeled probe thereof provided by the invention have excellent in-vivo biological performance, have excellent focus uptake in a focus with high expression of PSMA protein, and are long in residence time; compared with the prior art, the PSMA can be rapidly removed in non-target organs such as normal tissues, and the signal background is low, so that the PSMA has the characteristic of high target / non-target ratio, is particularly suitable for nuclide treatment and imaging of tumors, can improve the targeted PSMA nuclide diagnosis and treatment effect, and has the potential of clinical popularization and application.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and specifically to a PSMA-targeted small molecule compound and a preparation method and application thereof, a radionuclide-labeled probe and a preparation method and application thereof, a pharmaceutically acceptable form of the PSMA-targeted small molecule compound and the radionuclide-labeled probe and applications thereof, a pharmaceutical composition and applications thereof, and a kit and applications thereof. Background Art

[0002] The expression of prostate-specific membrane antigen (PSMA) is highly positively correlated with the progression of prostate cancer. It is overexpressed on the surface of more than 90% of prostate cancer cells, and its expression level is higher in cancer cells of patients with advanced and castration-resistant prostate cancer. In normal tissues, it is only expressed in small amounts in salivary glands, lacrimal glands, proximal renal tubules, etc. Therefore, PSMA has become an ideal target for the diagnosis and treatment of many tumors including prostate cancer, with the advantages of high sensitivity and high specificity. Positron emission tomography (PET) and single photon emission computed tomography (SPECT) in the field of nuclear medicine molecular imaging have the advantages of high sensitivity, but both rely on radionuclide-labeled drugs. In recent years, radionuclide-labeled drugs based on PSMA targets have received widespread attention and made gratifying progress, providing strong support for the diagnosis and treatment of prostate cancer.

[0003] 111 In-labeled monoclonal antibody ProstaScint is the first PSMA-targeted prostate cancer radioactive diagnostic drug approved by the US FDA. However, due to its large size, it has a certain adverse effect on cell membrane permeability and internalization rate. Small molecule radioactive drugs have better cell membrane permeability, faster pharmacokinetic rate and are easy to synthesize, and have great potential in radionuclide targeted therapy for prostate cancer. Clinical and preclinical research results show that existing PSMA-targeted small molecule radioactive drugs (such as 68 Ga-PSMA-617, 177 Lu-PSMA-617, 18 PSMA-1007 (F-labeled) has a certain amount of non-target organ retention, especially in the kidneys, liver, and normal glands, which affects the diagnosis and treatment effect to a certain extent, and the retention time in the lesion is limited. Therefore, it is of great significance to develop new drugs that can reduce the retention of radioactive drugs in non-target tissues and increase the retention time in lesions. Summary of the invention

[0004] In view of this, the object of the present invention is to provide a PSMA-targeted small molecule compound and a preparation method and application thereof, a radionuclide-labeled probe and a preparation method and application thereof, a pharmaceutical composition and application thereof, a kit and application thereof. The PSMA-targeted small molecule compound and a pharmaceutically acceptable form thereof, a radionuclide-labeled probe and a pharmaceutically acceptable form thereof provided by the present invention all have PSMA-targeting properties, and have high target organ uptake, long retention time, low non-target organ background, and a high target / non-target ratio.

[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0006] The present invention provides a PSMA-targeted small molecule compound having a structure shown in Formula I:

[0007]

[0008] In the formula I, n is an integer from 0 to 5;

[0009] R1 includes any of the following structures:

[0010]

[0011] R2 may be present or absent, and when present, R2 includes any one of the following structures:

[0012]

[0013] In said R2, a, b and t are independently integers between 0 and 5;

[0014] R3 is a group to be labeled, including any one of the following chelating structures:

[0015]

[0016] The present invention also provides a method for preparing the PSMA-targeted small molecule compound described in the above technical solution, including method 1, method 2 or method 3;

[0017] The method 1 comprises the following steps: using an R3 active compound with an activated ester group or an acid anhydride group to perform a substitution reaction with compound 1 to form an amide bond, thereby obtaining the PSMA-targeted small molecule compound; the R3 active compound with an activated ester group or an acid anhydride group comprises any one of the following structures:

[0018]

[0019] The method 2 comprises the following steps: using an R3 active compound with an isothiocyanate group to perform a substitution reaction with compound 1 to form a thiourea bond, thereby obtaining the PSMA-targeted small molecule compound; the R3 active compound with an isothiocyanate group comprises any one of the following structures:

[0020]

[0021]

[0022] The method 3 comprises the following steps: using an R3 chelating agent with a carboxyl structure to perform an amidation reaction with compound 1 under condensation reagent conditions to generate an amide bond, and then removing the protecting group to obtain the PSMA-targeted small molecule compound; the R3 chelating agent with a carboxyl structure comprises any one of the following structures, wherein A is a carboxyl protecting group:

[0023]

[0024] R1, R2 and n in the compound 1 are the same as R1, R2 and n in the formula I.

[0025] The present invention also provides a radionuclide labeled probe having a structure shown in Formula II or Formula III:

[0026]

[0027] Wherein, the definitions of R1, R2 and n are the same as those of R1, R2 and n in Formula I; R4 is a nuclide labeling group obtained by a coordination reaction between the R3 to-be-labeled group in the PSMA-targeted small molecule compound described in the above technical solution and the nuclide; and R5 includes any one of the following structures:

[0028]

[0029] Wherein, y and z are independently integers between 0 and 5; X is carbon, hydrogen or nitrogen; R6 is 18 F. 123 I. 124 I. 125 I. 131 I or 211 At.

[0030] Preferably, the nuclides include 18 F. 43 Sc, 44 Sc, 47 Sc, 51 Cr, 55 Co. 57 Co. 62 Cu, 64 Cu,67 Cu, 67 Ga, 68 Ga, 72 As, 72 Se, 89 Zr, 86 Y. 89 Sr. 86 Y. 90 Y. 97 Such as 99m Tc, 105 Rh, 101m Rh, 109 Pd, 111 In, 119 Sb, 128 8. 139 La, 140 La, 142 Pr, 149 Pm, 149 Tb, 151 Tb, 151 Eu, 153 Eu, 169 Eu, 153 Sm, 152 Gd, 153 Gd, 157 Gd, 159 Gd, 161 Tb, 165 Dy, 166 Ho, 169 2. 175 Yb, 177 Lu, 186 Re, 188 Re, 197 Hg, 198 Au, 201 Tl, 203 Pb, 211 At 212 Pb, 212 Bi, 213 Bi, 223 Ra, 227 Th and 225 At least one of Ac.

[0031] The present invention also provides a method for preparing the nuclide-labeled probe described in the above technical solution.

[0032] The method for preparing the nuclide-labeled probe having the structure shown in Formula II comprises the following steps: performing a coordination reaction between the PSMA-targeted small molecule compound described in the above technical solution and a nuclide to obtain a nuclide-labeled probe having the structure shown in Formula II;

[0033] The method for preparing the nuclide-labeled probe having the structure shown in Formula III comprises the following steps: subjecting Compound 1 to an amidation reaction with an active compound to obtain a nuclide-labeled probe having the structure shown in Formula III; the active compound comprises Compound 2 or Compound 3;

[0034]

[0035] The present invention also provides a pharmaceutically acceptable form, including the pharmaceutically acceptable form of the PSMA-targeted small molecule compound described in the above technical solution and / or the pharmaceutically acceptable form of the radionuclide-labeled probe described in the above technical solution, wherein the pharmaceutically acceptable form includes one or more of a salt, a stereoisomer, a racemate, a hydrate and a solvate.

[0036] The present invention also provides a pharmaceutical composition, comprising an active component and a pharmaceutically acceptable excipient; the active component comprises one or more of the PSMA-targeted small molecule compound described in the above technical solution, the radionuclide-labeled probe described in the above technical solution, and the pharmaceutically acceptable form described in the above technical solution.

[0037] The present invention also provides a lyophilization kit, comprising a precursor compound and a pharmaceutically acceptable excipient, wherein the precursor compound comprises the PSMA-targeted small molecule compound described in the above technical solution and one or more of the pharmaceutically acceptable forms of the PSMA-targeted small molecule compound described in the above technical solution.

[0038] The present invention also provides the use of the PSMA-targeted small molecule compound described in the above technical solution, the radionuclide-labeled probe described in the above technical solution, the pharmaceutically acceptable form described in the above technical solution, the pharmaceutical composition described in the above technical solution, or the lyophilized kit described in the above technical solution in the preparation of therapeutic agents, therapeutic drugs, diagnostic agents or diagnostic drugs for diseases mediated by PSMA protein.

[0039] Preferably, the PSMA protein-mediated disease includes tumors and / or metastases thereof;

[0040] The tumor includes one or more of prostate cancer, breast cancer, ovarian cancer, liver cancer, lung cancer, colorectal cancer, bone tissue sarcoma, connective tissue sarcoma, renal cell carcinoma, gastric cancer, pancreatic cancer, nasopharyngeal carcinoma, head and neck cancer, neuroendocrine tumors, skin melanoma and bone metastasis.

[0041] The present invention introduces a quinoline or naphthalene structure and a linker R2 between the targeting group and the coordination structure R3, which can adjust the distance and molecular structure between the targeting group and the coordination structure, and reduce the influence of the coordination structure on the activity of the compound. At the same time, the structure changes the physicochemical properties and pharmacokinetic properties of the nuclide-labeled probe by connecting with different types of nuclide chelating groups, accelerates the clearance rate of PSMA-targeted small molecule compounds or nuclide-labeled probes in non-target tissues, improves the uptake of target molecules and / or probes in PSMA-highly expressed lesions, and increases the target / non-target ratio. Compared with the existing PSMA-targeted nuclide-labeled probes, the nuclide-labeled probes provided by the present invention have excellent metabolic kinetic properties and high lesion uptake and retention time, a high target / non-target ratio, and excellent diagnostic and therapeutic effects on PSMA protein-mediated diseases, and have the potential to become diagnostic and therapeutic reagents and / or drugs that can be used for clinical applications and have commercial promotion value. As shown in the test results of the embodiments, the nuclide-labeled probes provided by the present invention have excellent metabolic kinetic properties and high lesion uptake and retention time, a high target / non-target ratio, and excellent diagnostic and therapeutic effects on PSMA protein-mediated diseases. It has the potential to become a diagnostic and therapeutic reagent and / or drug that can be used for clinical applications and has commercial promotion value. As shown in the test results of the embodiments, the nuclide-labeled probes provided by the present invention 68 The absolute value of uptake of Ga-labeled radionuclide-labeled probe in tumors is 68 The activity of Ga-PSMA-617 is twice that of Ga-PSMA-617, which is a very promising radionuclide diagnostic probe. In addition, by adjusting the appropriate specific activity or drug combination, it is helpful to obtain a better target / non-target ratio and enhance the uptake of radionuclide labeled probes in tumors.

[0042] The PSMA-targeted small molecule compound provided by the present invention can chelate with the nuclide through the labeled group R3 to form a PSMA-targeted nuclide labeled probe with high affinity and high specificity, which has the characteristics of convenient labeling, strong labeling ability, short labeling time, high labeling yield, etc. The prepared nuclide labeled probe has good stability, high sensitivity, excellent pharmacokinetic properties, high lesion uptake, and high target / non-target ratio. It can be applied to the labeling of a variety of diagnostic and therapeutic nuclides, and can also be used to construct a diagnostic and therapeutic platform based on diagnostic and therapeutic nuclide pairs, and has a good application prospect in the preparation of therapeutic reagents and / or drugs, diagnostic reagents and / or drugs for PSMA protein-mediated diseases. It is conducive to the application and clinical promotion of nuclide labeled probes.

[0043] The preparation method of the PSMA-targeted small molecule compound provided by the present invention has simple synthesis, simple operation, low production cost, and is suitable for industrial production.

[0044] The radionuclide labeled probe provided by the present invention has the characteristics of convenient labeling, strong labeling ability, short labeling time, high labeling yield, etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0045] Figure 1 This is the mass spectrum identification diagram of compound DNB8P1;

[0046] Figure 2 This is the mass spectrometry identification diagram of compound DNA8P1;

[0047] Figure 3 This is the mass spectrum identification diagram of compound DNC8P1;

[0048] Figure 4 This is the mass spectrum identification diagram of compound DND8P1;

[0049] Figure 5 This is the mass spectrometry identification diagram of compound DNA6P1;

[0050] Figure 6 This is the mass spectrum identification diagram of compound DNB6P1;

[0051] Figure 7 This is the mass spectrum identification diagram of compound DCA8P1;

[0052] Figure 8 This is the mass spectrum identification diagram of compound DCB8P1;

[0053] Fig. 9 This is the mass spectrum identification diagram of compound DNB8P2;

[0054] Fig.10 This is the mass spectrum identification diagram of compound DCB8P2;

[0055] Fig.11 This is the mass spectrum identification diagram of compound NNA8P1;

[0056] Fig.12 This is the mass spectrum identification diagram of compound NCB8P1;

[0057] Fig.13 This is the mass spectrum identification diagram of compound NNB8P1;

[0058] Fig.14 This is the mass spectrum identification diagram of compound DGNB8P1;

[0059] Fig.15 This is the mass spectrum identification diagram of compound DSNB8P1;

[0060] Fig.16 This is the mass spectrum identification diagram of compound DGNE8P1;

[0061] Fig.17 This is the mass spectrum identification diagram of compound DGNF8P1;

[0062] Fig.18 HPLC charts of compounds DNA8P1 and DNB8P1;

[0063] Fig.19 HPLC charts of compounds DNC8P1 and DND8P1;

[0064] Fig. 20HPLC charts of compounds DNA6P1 and DNB6P1;

[0065] Fig.21 HPLC charts of compounds DCA8P1 and DCB8P1;

[0066] Fig. 22 HPLC charts of compounds DNB8P2 and DCB8P2;

[0067] Fig.23 HPLC charts of compounds NNA8P1, NNB8P1 and NCB8P1;

[0068] Fig.24 HPLC charts of compounds DGNB8P1 and DSNB8P1;

[0069] Fig.25 HPLC charts of compounds DGNE8P1 and DGNF8P1;

[0070] Fig.26 For the probe 68 Ga]Ga-DNA8P1 and [ 68 HPLC profile of Ga]Ga-DNB8P1;

[0071] Fig. 27 For the probe 68 Ga]Ga-DNC8P1 and [ 68 HPLC profile of Ga]Ga-DND8P1;

[0072] Fig.28 For the probe 68 Ga]Ga-DCA8P1 and [ 68 HPLC profile of Ga]Ga-DCB8P1;

[0073] Fig.29 For the probe 68 Ga]Ga-DNA6P1 and [ 68 HPLC profile of Ga]Ga-DNB6P1;

[0074] Fig.30 For the probe 68 Ga]Ga-DNB8P2 and [ 68 HPLC profile of Ga]Ga-DCB8P2;

[0075] Fig.31 For the probe 68 Ga]Ga-DGNB8P1 and [ 68 HPLC profile of Ga]Ga-DSNB8P1;

[0076] Fig.32For the probe 68 Ga]Ga-DGNE8P1 and [ 68 HPLC profile of Ga]Ga-DGNF8P1;

[0077] Fig.33 For the probe 18 F]AlF-NNA8P1, [ 18 F]AlF-NNB8P1 and [ 18 F] HPLC profile of AlF-NCB8P1;

[0078] Fig.34 For the probe 177 Lu]Lu-DGNB8P1, [ 177 Lu]Lu-DSNB8P1, [ 177 Lu]Lu-DGNE8P1 and [ 177 HPLC profile of Lu]Lu-DGNF8P1;

[0079] Fig.35 For the probe 44 Sc]Sc-DNB8P1 and [ 44 Sc] HPLC profile of Sc-DGNB8P1;

[0080] Fig.36 For probe 131 HPLC profile of IPB-NB8P1;

[0081] Fig.37 For the probe 68 HPLC graph of the stability of Ga]Ga-DNA8P1 in physiological saline and serum;

[0082] Fig.38 For the probe 68 HPLC graph of the stability of Ga]Ga-DNB8P1 in normal saline and serum;

[0083] Fig.39 For the probe 68 HPLC graph of the stability of Ga]Ga-DNA6P1 in physiological saline and serum;

[0084] Fig.40 For the probe 68 HPLC graph of the stability of Ga]Ga-DNB6P1 in normal saline and serum;

[0085] Fig.41 For the probe 68 HPLC graph of the stability of Ga]Ga-DNC8P1 in physiological saline and serum;

[0086] Fig.42For the probe 68 HPLC graph of the stability of Ga]Ga-DND8P1 in normal saline and serum;

[0087] Fig.43 For the probe 68 HPLC graph of the stability of Ga]Ga-DCA8P1 in physiological saline and serum;

[0088] Fig.44 For the probe 68 HPLC graph of the stability of Ga]Ga-DCB8P1 in normal saline and serum;

[0089] Fig.45 For the probe 18 F] HPLC chart of the stability of AlF-NNA8P1 in normal saline and serum;

[0090] Fig.46 For the probe 18 F] HPLC chart of the stability of AlF-NNB8P1 in normal saline and serum;

[0091] Fig.47 For the probe 18 F] HPLC chart of the stability of AlF-NCB8P1 in normal saline and serum;

[0092] Fig.48 For the probe 177 HPLC chart of the stability of Lu]Lu-DGNB8P1 in normal saline and serum;

[0093] Fig.49 For the probe 177 HPLC graph of the stability of Lu]Lu-DGNF8P1 in normal saline and serum;

[0094] Fig.50 For the probe 44 HPLC chart of the stability of Sc]Sc-DNB8P1 in normal saline and serum;

[0095] Fig.51 For the probe 44 Sc] Sc-DGNB8P1 stability HPLC chart in normal saline and serum;

[0096] Fig.52 For the probe 68 Ga]Ga-DNA8P1 and [ 68 The K of Ga]Ga-DNB8P1 in PSMA-positive cells PC3-PIP d Test curve;

[0097] Fig.53 For the probe 68 Ga]Ga-DNC8P1 and [ 68 The K of Ga]Ga-DND8P1 in PSMA-positive cells PC3-PIP d Test curve;

[0098] Fig.54 For the probe 68 Ga]Ga-DNA6P1 and [ 68 The K of Ga]Ga-DNB6P1 in PSMA-positive cells PC3-PIP d Test curve;

[0099] Fig.55 For the probe 68 Ga]Ga-DCA8P1 and [ 68 The K of Ga]Ga-DCB8P1 in PSMA-positive cells PC3-PIP d Test curve;

[0100] Fig.56 For the probe 18 F]AlF-NNA8P1, [ 18 F]AlF-NNB8P1 and [ 18 F]AlF-NCB8P1 K in PSMA-positive cells PC3-PIP d Test curve;

[0101] Fig.57 For the probe 18 F]AlF-NNA8P1, [ 18 F]AlF-NNB8P1 and [ 18 F] The cell uptake results and cell inhibition results of AlF-NCB8P1 in PSMA-positive cells PC3-PIP at different times (60 min time point) were compared with the cell uptake results of the probe in PSMA-negative cells PC3 at 60 min;

[0102] Fig.58 For the probe 177 Lu]Lu-DGNB8P1 and [ 177 The cell uptake results and cell inhibition results of Lu]Lu-DSNB8P1 in PSMA-positive cells PC3-PIP at different times (2h and 4h time points) were compared with the cell uptake results of the probe in negative cells PC3 at 2h and 4h;

[0103] Fig.59 For the probe 68 PET imaging results of Ga]Ga-DNA8P1 (A) and uptake values ​​of major tissues and target / non-target ratio (B);

[0104] Fig.60 For the probe 68 PET imaging results of Ga]Ga-DNB8P1 (A) and uptake values ​​of major tissues and target / non-target ratio (B);

[0105] Fig.61 For the probe 68 PET imaging results of Ga]Ga-DNC8P1 (A) and uptake values ​​of major tissues and target / non-target ratio (B);

[0106] Fig.62 For the probe 68 PET imaging results of Ga]Ga-DND8P1 (A) and uptake values ​​of major tissues and target / non-target ratio (B);

[0107] Fig.63 For the probe 68 PET imaging results of Ga]Ga-DNA6P1 (A) and uptake values ​​of major tissues and target / non-target ratio (B);

[0108] Fig.64 For the probe 68 PET imaging results of Ga]Ga-DNB6P1 (A) and uptake values ​​of major tissues and target / non-target ratio (B);

[0109] Fig.65 For the probe 68 PET imaging results of Ga]Ga-DCA8P1 (A) and uptake values ​​of major tissues and target / non-target ratio (B);

[0110] Fig.66 For the probe 68 PET imaging results of Ga]Ga-DCB8P1 (A) and uptake values ​​of major tissues and target / non-target ratio (B);

[0111] Fig.67 For the probe 68 PET imaging results of Ga]Ga-DNB8P2 (A) and uptake values ​​of major tissues and target / non-target ratio (B);

[0112] Fig.68 For the probe 68 PET imaging results of Ga]Ga-DCB8P2 (A) and uptake values ​​of major tissues and target / non-target ratio (B);

[0113] Fig.69 For the probe 68 PET imaging results of Ga]Ga-DGNB8P1 (A) and uptake values ​​of major tissues and target / non-target ratio (B);

[0114] Fig.70 For the probe 68 PET imaging results of Ga]Ga-DSNB8P1 (A) and uptake values ​​of major tissues and target / non-target ratio (B);

[0115] Fig.71 For the probe 68 PET imaging results of Ga]Ga-DGNE8P1 (A) and uptake values ​​of major tissues and target / non-target ratio (B);

[0116] Fig.72 For the probe 68 PET imaging results of Ga]Ga-DGNF8P1 (A) and uptake values ​​of major tissues and target / non-target ratio (B);

[0117] Fig.73 For different contents of competitive binders (DNB8P1, nat Ga-DNB8P1, PSMA-617) in the presence of probe [ 68 Comparison of PET imaging results (A) and uptake values ​​of major tissues (B) of Ga]Ga-DNB8P1;

[0118] Fig.74 For the probe 68 Ga]Ga-DNB8P1 and classical probes [ 68 PET imaging results of Ga]Ga-PSMA-617 (A), comparison of uptake values ​​of major tissues (B), and comparison of target / non-target ratios (C);

[0119] Fig.75 For the probe 18 F] PET imaging results of AlF-NNA8P1 (A) and uptake values ​​of major tissues (B);

[0120] Fig.76 For the probe 18 F] PET imaging results of AlF-NNB8P1 (A) and uptake values ​​of major tissues (B);

[0121] Fig.77 For the probe 18 F] PET imaging results of AlF-NCB8P1 (A) and uptake values ​​of major tissues (B);

[0122] Fig.78 For the probe 44 Sc]Sc-DGNB8P1 PET imaging results (A) and comparison of major tissue uptake values ​​and target / non-target ratios (B);

[0123] Fig.79 For the probe177 SPECT imaging results (A) and target / non-target ratio (B) of Lu]Lu-DGNB8P1;

[0124] Fig.80 For the probe 177 SPECT imaging results (A) and target / non-target ratio (B) of Lu]Lu-DSNB8P1;

[0125] Fig.81 For the probe 177 SPECT imaging results (A) and target / non-target ratio (B) of Lu]Lu-DGNF8P1;

[0126] Fig.82 For the probe 177 SPECT imaging results of Lu]Lu-PSMA-617;

[0127] Fig.83 In the presence of 20 μg of competitive binding agent DGNB8P1, the probe 177 SPECT imaging results (A) and target / non-target ratio (B) of Lu]Lu-DGNB8P1;

[0128] Fig.84 For the probe 18 F] Biodistribution results of AlF-NNA8P1 in tumor-bearing mice;

[0129] Fig.85 For the probe 18 F] Biodistribution results of AlF-NCB8P1 in tumor-bearing mice;

[0130] Fig.86 For the probe 177 The biodistribution results of Lu]Lu-DGNB8P1 in tumor-bearing mice;

[0131] Fig.87 For the probe 44 Sc]Sc-DGNB8P1 biodistribution results in tumor-bearing mice;

[0132] Fig.88 for[ 177 Lu]Lu-PSMA-617 probe (18.5MBq) and different injection doses of [ 177 Tumor inhibition results of Lu]Lu-DGNB8P1 probe in tumor-bearing mice. DETAILED DESCRIPTION

[0133] The present invention provides a PSMA-targeted small molecule compound having a structure shown in Formula I:

[0134]

[0135] In the present invention, n in Formula I is an integer of 0 to 5. In a specific embodiment, n can be 0, 1, 2, 3, 4 or 5.

[0136] In the present invention, R1 in Formula I includes any one of the following structures:

[0137]

[0138] In the present invention, R2 in the formula I may be present or absent, and when R2 is present, it includes any one of the following structures:

[0139]

[0140] Wherein, a, b and t are independently integers ranging from 0 to 5. In a specific embodiment, a, b and t can be independently 0, 1, 2, 3, 4 or 5.

[0141] In the present invention, R3 in Formula I is a group to be labeled, including any one of the following structures:

[0142]

[0143]

[0144] In the present invention, the -CO-terminus in R1 is connected to the -NH-terminus of the lysine residue in Formula I to form an amide bond; when R2 is not present, the -NH-terminus in R1 is directly connected to R3; when R2 is present, the -NH-terminus in R1 is connected to the -CO-terminus in R2, and the -NH-terminus in R2 is further connected to R3.

[0145] In the present invention, the PSMA targeting small molecule compound preferably has any one of the structures shown in Formula I-1 to Formula I-10, in which n is independently 1 or 2, Q is independently N or CH, and R2 is independently absent or is a different linking group. The PSMA targeting small molecule compounds having the structures shown in Formula I-1 to Formula I-10 and their names are shown in Tables 1 to 10.

[0146]

[0147] Table 1 PSMA-targeted small molecule compounds having the structure shown in Formula I-1 and their names

[0148]

[0149] Table 2 PSMA-targeted small molecule compounds having the structure of formula I-2 and their names

[0150]

[0151]

[0152] Table 3 Names of PSMA-targeted small molecule compounds having the structure of Formula I-3

[0153]

[0154]

[0155] Table 4 PSMA-targeted small molecule compounds having the structure shown in Formula I-4 and their names

[0156]

[0157]

[0158] Table 5 Names of PSMA-targeted small molecule compounds having the structure of Formula I-5

[0159]

[0160]

[0161] Table 6 Names of PSMA-targeted small molecule compounds with the structure shown in Formula I-6 and their names

[0162]

[0163]

[0164] Table 7 PSMA-targeted small molecule compounds having the structure shown in Formula I-7 and their names

[0165]

[0166]

[0167] Table 8 PSMA-targeted small molecule compounds having the structure shown in Formula I-8 and their names

[0168]

[0169]

[0170] Table 9 PSMA-targeted small molecule compounds having the structure shown in Formula I-9 and their names

[0171]

[0172]

[0173] Table 10 PSMA-targeted small molecule compounds having the structure shown in Formula I-10 and their names

[0174]

[0175]

[0176] The results of clinical and preclinical studies show that existing PSMA-targeted radioactive drugs have a certain amount of non-target organ retention, especially retention in the kidneys, liver, and normal glands, which affects the diagnosis and treatment effect to a certain extent. Moreover, the existing drugs can detect a limited number of lesions, and there is a possibility of missed detection for some micro-metastatic lesions. It is necessary to develop drugs that can detect more lesions; the existing therapeutic radioactive drugs have a limited retention time in lesions and require a large dose. Therefore, the development of radionuclide-targeted drugs with higher lesion uptake levels, better therapeutic effects, and better target / non-target ratios can improve the therapeutic effect, while reducing the patient's dosage or frequency of administration and reducing the possibility of adverse reactions. The PSMA-targeted small molecule compound provided by the present invention is easy to label, has good stability, high sensitivity, excellent pharmacokinetic properties, high lesion uptake, and a high target / non-target ratio, and is of great value as a radioactive drug.

[0177] The present invention provides a method for preparing the PSMA-targeted small molecule compound described in the above technical solution, including method 1, method 2 or method 3.

[0178] In the present invention, the method 1 comprises the following steps: using an R3 active compound with an activated ester group or an acid anhydride group to carry out a substitution reaction with compound 1 to form an amide bond, thereby obtaining the PSMA-targeted small molecule compound; the R3 active compound with an activated ester group or an acid anhydride group comprises any one of the following structures:

[0179]

[0180]

[0181] The definitions of R1, R2 and n in the compound 1 are the same as those of R1, R2 and n in the formula I.

[0182] Unless otherwise specified, the materials and equipment used in the present invention are all commercially available products in the art.

[0183] In the present invention, the molar ratio of the compound 1 to the R3 active compound with an activated ester group or anhydride group is preferably 1:1-10, and in specific embodiments, it can be 1:1, 1:1.2, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 or 1:10. In the present invention, the solvent used in the substitution reaction in the method 1 preferably includes one or more of an organic solvent, water and PBS (phosphate buffer solution). In the present invention, the organic solvent preferably includes one or more of N-methylpyrrolidone (NMP), dichloromethane (DCM), dimethyl sulfoxide (DMSO) and N,N-dimethylformamide (DMF). The pH value of the PBS is preferably 7-12, and in specific embodiments, it can be 7, 8, 9, 10, 11 or 12. The present invention has no special limitation on the amount of the solvent, as long as it can ensure the smooth progress of the substitution reaction.

[0184] In the present invention, the substitution reaction is preferably carried out in the presence of an alkaline agent; the alkaline agent is preferably an organic base, more preferably triethylamine (TEA) and / or N,N-diisopropylethylamine (DIPEA). In the present invention, the molar ratio of the compound 1 to the alkaline agent is preferably 1:0.1 to 1000, and in specific embodiments can be 1:0.1, 1:0.5, 1:1, 1:2, 1:3, 1:4, 1:1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:15, 1:20, 1:50, 1:100, 1:200, 1:300, 1:400, 1:500, 1:600, 1:700, 1:800, 1:900 or 1:1000.

[0185] In the present invention, the temperature of the substitution reaction is preferably 15-60°C, and in specific embodiments it can be 15°C, 20°C, 30°C, 40°C, 50°C or 60°C; the time of the substitution reaction is preferably 0.5-48h, and in specific embodiments it can be 0.5h, 1h, 2h, 5h, 10h, 15h, 20h, 25h, 30h, 35h, 40h, 45h or 48h.

[0186] After completing the substitution reaction, the present invention preferably further comprises: subjecting the obtained substitution reaction solution to HPLC purification and then freeze-drying to obtain the PSMA-targeted small molecule compound. The present invention has no special restrictions on the temperature and time of the freeze-drying, and freeze-drying to constant weight (i.e., freeze-drying) is sufficient. The present invention has no special restrictions on the HPLC purification conditions, and liquid phase purification conditions that can obtain compound purity that meets the requirements are sufficient.

[0187] In the present invention, the method 2 comprises the following steps: using an R3 active compound with an isothiocyanate group to carry out a substitution reaction with compound 1 to form a thiourea bond, thereby obtaining the PSMA-targeted small molecule compound; the R3 active compound with an isothiocyanate group is as follows:

[0188]

[0189] In the present invention, the molar ratio of the compound 1 to the isothiocyanate group R3 active compound is preferably 1:1 to 10, and in specific embodiments can be 1:1, 1:1.2, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 or 1:10. In the present invention, the substitution reaction conditions and post-treatment methods of the method 2 are the same as those of the method 1, and will not be described one by one here.

[0190] In the present invention, the method 3 comprises the following steps: using an R3 chelating reagent with a carboxyl structure to perform an amidation reaction with compound 1 under condensation reagent conditions to form an amide bond, and then removing the protecting group to obtain the PSMA-targeted small molecule compound; the R3 chelating reagent with a carboxyl structure is shown below and includes any one of the following structures:

[0191]

[0192] Wherein, A is a carboxyl protecting group, preferably including a methyl ester group, an ethyl ester group or a tert-butyl ester group; the definitions of R1, R2 and n in the compound 1 are the same as those of R1, R2 and n in the formula I.

[0193] In the present invention, the molar ratio of the compound 1 to the R3 chelating agent having a carboxyl structure is preferably 1:1-10, and in specific embodiments can be 1:1, 1:1.2, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 or 1:10.

[0194] In the present invention, the amidation reaction is preferably carried out in the presence of a condensation reagent, an organic amine and a solvent. In the present invention, the condensation reagent preferably includes one or more of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), O-(benzotriazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU), dicyclohexylcarbodiimide (DCC), N,N-diisopropylcarbodiimide (DIC) and carbodiimide hydrochloride (EDC), oxalyl chloride, thionyl chloride, phosphorus oxychloride, carbon disulfide and succinimide activated ester; the molar ratio of the compound 1 to the condensation reagent is preferably 1:0.5-10, and in specific embodiments, it can be 1:0.5, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 or 1:10. In the present invention, the organic amine preferably includes N,N-diisopropylethylamine (DIPEA) and / or triethylamine (Et3N); the molar ratio of the compound 1 to the organic amine is preferably 1:0.5-10, and in specific embodiments can be 1:0.5, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 or 1:10. In the present invention, the solvent preferably includes one or more of an organic solvent and / or water and / or PBS (phosphate buffer solution). In the present invention, the organic solvent preferably includes one or more of N-methylpyrrolidone (NMP), dichloromethane (DCM), dimethyl sulfoxide (DMSO) and N,N-dimethylformamide (DMF); the present invention has no special limitation on the amount of the organic solvent, as long as it can ensure the smooth progress of the amidation reaction. In the present invention, the temperature of the amidation reaction is preferably room temperature, and the time is preferably 0.5 to 48 hours. In specific embodiments, it can be 0.5 hours, 1 hour, 2 hours, 5 hours, 10 hours, 15 hours, 20 hours, 25 hours, 30 hours, 35 hours, 40 hours, 45 hours or 48 hours.

[0195] In the present invention, the deprotection reagent used for removing the protecting group preferably includes a strong acid or a strong base, more preferably trifluoroacetic acid (TFA) or piperidine; the present invention has no special limitation on the concentration of the deprotection reagent solution, as long as the deprotection is carried out smoothly. In the present invention, the temperature for removing the protecting group is preferably 0 to 100°C, and in specific embodiments, it can be 0°C, 10°C, 20°C, 30°C, 40°C, 50°C, 60°C, 70°C, 80°C, 90°C or 100°C; the time for removing the protecting group is preferably 1min to 24h, and in specific embodiments, it can be 1min, 5min, 30min, 1h, 5h, 10h, 15h, 20h or 24h.

[0196] After the removal of the protecting group, the present invention preferably further comprises: subjecting the obtained reaction solution of the removing protecting group to high performance liquid chromatography purification and then freeze drying to obtain the PSMA targeting small molecule compound. The present invention has no special restrictions on the temperature and time of the freeze drying, and freeze drying to constant weight (i.e., freeze drying) is sufficient. The present invention has no special restrictions on the high performance liquid chromatography purification conditions, and the liquid phase purification conditions that can obtain the compound purity meeting the requirements are sufficient.

[0197] The present invention provides a radionuclide labeled probe having a structure shown in Formula II or Formula III:

[0198]

[0199] The definitions of R1, R2 and n in Formula II and Formula III are the same as those of R1, R2 and n in Formula I; R4 is a nuclide labeling group, which is obtained by the coordination reaction between the R3 to-be-labeled group in the PSMA-targeted small molecule compound described in the above technical solution and the nuclide; R5 includes any one of the following structures:

[0200]

[0201] Wherein, y and z are independently integers between 0 and 5, and in a specific embodiment, can be 0, 1, 2, 3, 4 or 5; X is carbon, hydrogen or nitrogen; R6 is 18 F. 123 I. 124 I. 125 I. 131 I or 211 At.

[0202] In the present invention, the nuclides preferably include 18 F. 43 Sc, 44 Sc, 47 Sc, 51 Cr, 55 Co. 57 Co. 62 Cu, 64 Cu, 67 Cu, 67 Ga, 68 Ga, 72 As, 72 Se, 89 Zr, 86 Y. 89 Sr. 86 Y. 90 Y. 97 Such as 99m Tc, 105 Rh, 101m Rh,109 Pd, 111 In, 119 Sb, 128 8. 139 La, 140 La, 142 Pr, 149 Pm, 149 Tb, 151 Tb, 151 Eu, 153 Eu, 169 Eu, 153 Sm, 152 Gd, 153 Gd, 157 Gd, 159 Gd, 161 Tb, 165 Dy, 166 Ho, 169 2. 175 Yb, 177 Lu, 186 Re, 188 Re, 197 Hg, 198 Au, 201 Tl, 203 Pb, 211 At 212 Pb, 212 Bi, 213 Bi, 223 Ra, 227 Th and 225 At least one of Ac.

[0203] In the present invention, the formula III preferably has any one of the structures shown in formula III-1 to formula III-6, in which Q is independently N or CH, and R2 is independently absent or is a different connecting group. The nuclide-labeled probes having the structures shown in formula III-1 to formula III-6 and their names are shown in Tables 11 to 16, in which / represents or, for example 123 / 124 / 125 / 131 I stands for 123 I. 124 I. 125 I or 131 I.

[0204]

[0205] Table 11 Nuclide-labeled probes with the structure shown in Formula III-1 and their names

[0206]

[0207]

[0208] Table 12 Radionuclide labeled probes having the structure shown in Formula III-2 and their names

[0209]

[0210]

[0211] Table 13 Radionuclide labeled probes with the structure shown in Formula III-3 and their names

[0212]

[0213]

[0214] Table 14 Nuclide-labeled probes with the structure shown in Formula III-4 and their names

[0215]

[0216] Table 15 Nuclide labeled probes with the structure shown in Formula III-5 and their names

[0217]

[0218]

[0219] Table 16 Nuclide labeled probes with the structure shown in Formula III-6 and their names

[0220]

[0221]

[0222]

[0223] The present invention provides a method for preparing the nuclide-labeled probe described in the above technical solution, comprising the following steps: performing a coordination reaction between the PSMA-targeted small molecule compound and a nuclide to obtain a nuclide-labeled probe having a structure shown in Formula II.

[0224] In the present invention, the nuclide-labeled probe having the structure shown in Formula II is preferably prepared by a wet labeling method or a freeze-drying labeling method.

[0225] In the present invention, the wet labeling method for preparing a nuclide labeled probe preferably includes the following steps: mixing a solution containing a precursor compound with a nuclide solution, performing a coordination reaction and then diluting to obtain a reaction solution containing a nuclide labeled probe; the precursor compound is the PSMA-targeted small molecule compound described in the above technical solution or a pharmaceutically acceptable form of the PSMA-targeted small molecule compound described in the above technical solution.

[0226] The present invention has no special limitation on the solvent in the solution of the PSMA-targeted small molecule compound, and any solvent familiar to those skilled in the art can be used, which may be one or more of a buffer solution, water, and an organic solvent; the buffer solution preferably includes PBS, acetic acid-acetate solution, or aluminum chloride-acetate solution; the organic solvent preferably includes one or more of acetonitrile, ethanol, DMSO, and DMF. In the present invention, when the nuclide is 18 F, the solvent preferably includes acetic acid-acetate solution or aluminum chloride-acetate solution; when the nuclide is 68 Ga or 177 Lu, the solvent preferably includes acetic acid-acetate solution. In the present invention, the concentration of the solution of the PSMA targeting small molecule compound is preferably 0.001-1000 mg / mL, more preferably 0.01-1 mg / mL. In the present invention, the ratio of the mass of the PSMA targeting compound to the radioactivity of the labeled nuclide in the labeled nuclide solution is preferably 20-400 μg: 1 kBq-1000 GBq, more preferably 20-400 μg: 0.037-74000 MBq, further preferably 20-200 μg: 0.037-7400 MBq. The present invention has no special limitation on the nuclide solution, and a nuclide solution well known to those skilled in the art may be used, such as 68 GaCl3 hydrochloric acid solution, 18 F - Solution, 177 LuCl3 solution or Na 99m TcO4 eluent, 68 The GaCl3 hydrochloric acid solution is preferably obtained by eluting from a germanium gallium generator. 99m The TcO4 eluent is preferably obtained by elution from a molybdenum-technetium generator.

[0227] In the present invention, the temperature of the coordination reaction is preferably 25 to 100°C, more preferably 80 to 100°C, and the time of the coordination reaction is preferably 10 to 60 min, more preferably 20 to 30 min. When the temperature of the coordination reaction is higher than room temperature, the present invention preferably further comprises cooling the obtained coordination reaction system to room temperature after the coordination reaction, and can be diluted according to actual needs, and aseptically filtered to obtain a radionuclide labeled probe solution. The present invention has no special limitation on the cooling, and a cooling method familiar to those skilled in the art can be adopted, such as natural cooling. In the present invention, the dilution preferably utilizes physiological saline or water for injection. In the present invention, the concentration of the radionuclide labeled probe solution is preferably 0.037 to 3700 MBq / mL.

[0228] In the present invention, when the nuclide is 18F, the coordination reaction is preferably carried out under the condition of a mixed medium containing a co-coordinated ion, a buffer solution and an organic solvent; the co-coordinated ion is preferably a metal ion, more preferably an aluminum ion; the buffer solution preferably includes an aluminum chloride-acetate solution; and the organic solvent preferably includes acetonitrile.

[0229] In the present invention, when the nuclide is 99m When Tc, the coordination reaction is preferably carried out in the presence of a reducing agent and a coordinating ligand, the reducing agent preferably includes SnCl2 or triphenylphosphine tris-sulfonic acid sodium salt (TPPTS), and the SnCl2 is preferably used in the form of a SnCl2 hydrochloric acid solution; the concentration of the hydrochloric acid solution is preferably 0.00001-12 mol / L. For each labeling, the amount of SnCl2 used is preferably 0.001-100 mg, more preferably 0.01-0.2 mg. In the present invention, the synergistic ligand preferably includes at least one of N-tris(hydroxymethyl)methylglycine (Tricine), triphenylphosphine trissulfonic acid sodium salt (TPPTS), ethylenediamine-N,N'-diacetic acid (EDDA), ethylenediaminetetraacetic acid (EDTA), and 3-diphenylphosphinobenzenesulfonic acid sodium salt (TPPMS); the mass ratio of the PSMA targeting small molecule compound to the synergistic ligand is preferably 0.01-0.5:0.1-50, and more preferably 0.02-0.1:1-10.

[0230] In the present invention, the preparation of the radionuclide labeled probe by the freeze-drying labeling method preferably includes the following steps: freeze-drying the solution containing the PSMA-targeted small molecule compound and then sealing it to obtain a freeze-dried medicine box; adding a solvent to the freeze-dried medicine box to dissolve it, and then adding a radionuclide solution to carry out coordination reaction and then dilute it to obtain a radionuclide labeled probe solution. In the present invention, the freeze-drying preferably includes dispensing the solution containing the PSMA-targeted small molecule compound into a freeze-drying container and then freeze-drying it; the present invention does not specifically limit the freeze-drying conditions, and the freeze-drying conditions familiar to those skilled in the art can be used.

[0231] The present invention preferably adds pharmaceutically acceptable excipients to the injection containing the radionuclide labeled probe obtained by the wet labeling method or the freeze-dried medicine box obtained by the freeze-drying labeling method as needed; the pharmaceutically acceptable excipients preferably include at least one of excipients, antioxidants, adhesives, buffers, colorants, diluents, disintegrants, emulsifiers, flavoring agents, glidants, lubricants, preservatives, stabilizers, surfactants, tablets and wetting agents; the present invention does not specifically limit the types of the above-mentioned pharmaceutically acceptable excipients, and pharmaceutically acceptable excipients well known to those skilled in the art can be used.

[0232] In the present invention, when the radiochemical purity of the nuclide labeled probe solution prepared by the wet labeling method or the freeze-drying labeling method is lower than 95%, it is preferred to further include purifying the nuclide labeled probe solution; the purification preferably includes column separation purification, and the chromatographic column used in the column separation purification preferably includes a Sep-Pak C18 separation column, and the Sep-Pak C18 separation column is preferably activated and eluted with anhydrous ethanol and water in sequence before use. In the present invention, the eluent used for the column separation purification is preferably water and anhydrous ethanol in sequence, and the eluate of anhydrous ethanol is collected and the solvent is removed, and then diluted to obtain a high-purity nuclide labeled probe solution. In the present invention, the dilution is preferably performed using physiological saline or water for injection. In the present invention, the radioactivity concentration of the high-purity nuclide labeled probe solution is preferably 0.037 to 3700 MBq / mL.

[0233] The present invention provides a method for preparing the nuclide-labeled probe described in the above technical solution, comprising the following steps: subjecting the compound 1 to an amidation reaction with an active compound to obtain a nuclide-labeled probe having a structure shown in formula III; the active compound comprises compound 2 or compound 3;

[0234]

[0235] Wherein, the definitions of y, z, X and R6 are the same as those of y, z, X and R6 in R5.

[0236] In the present invention, the molar ratio of the compound 1 to the active compound is preferably 1:0.0000001-10, and in specific embodiments can be 1:0.0000001, 1:0.000001, 1:0.00001, 1:0.0001, 1:0.001, 1:0.1, 1:0.5, 1:1, 1:1.5, 1:2, 1:2.5, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 or 1:10. In the present invention, the organic solvent used in the amidation reaction preferably includes one or more of an organic solvent and / or water and / or PBS (phosphate buffer solution). In the present invention, the organic solvent preferably includes one or more of N-methylpyrrolidone (NMP), dichloromethane (DCM), dimethyl sulfoxide (DMSO) and N,N-dimethylformamide (DMF). The present invention has no special limitation on the amount of the organic solvent, as long as the amidation reaction can proceed smoothly.

[0237] The preparation method provided by the present invention has the advantages of simple and easy labeling, good stability of the obtained radionuclide labeled probe, high tumor uptake, etc., and is suitable for industrial production and clinical promotion.

[0238] The present invention provides a pharmaceutically acceptable form of existence, including the pharmaceutically acceptable form of existence of the PSMA-targeted small molecule compound described in the above technical solution and / or the pharmaceutically acceptable form of existence of the nuclide-labeled probe described in the above technical solution, wherein the pharmaceutically acceptable form of existence includes one or more of salts, stereoisomers, racemates, hydrates and solvates. In the present invention, the salt preferably includes trifluoroacetate, phosphate, formate, acetate, potassium salt or sodium salt. The present invention has no special limitation on the tautomer, racemate, hydrate or solvate, and the tautomer, racemate, hydrate or solvate well known to those skilled in the art can be used. The present invention has no special limitation on the preparation method of the tautomer, racemate, hydrate, solvate or salt, and the preparation method of the tautomer, racemate, hydrate, solvate or salt well known to those skilled in the art can be used.

[0239] The present invention also provides a pharmaceutical composition, comprising an active ingredient and a pharmaceutically acceptable excipient; the active ingredient comprises one or more of the PSMA-targeted small molecule compound described in the above technical solution, the pharmaceutically acceptable form of the PSMA-targeted small molecule compound described in the above technical solution, the radionuclide labeled probe described in the above technical solution, and the pharmaceutically acceptable form of the radionuclide labeled probe described in the above technical solution. In the present invention, the optional types of the pharmaceutically acceptable excipients are the same as the aforementioned pharmaceutically acceptable excipients, and are not repeated here. In the present invention, when the active ingredient comprises a radionuclide labeled probe and / or a pharmaceutically acceptable form of the radionuclide labeled probe, the dosage form of the pharmaceutical composition is preferably an injection, preferably administered by intravenous injection; when the active ingredient does not contain a radionuclide labeled probe and / or a pharmaceutically acceptable form of the radionuclide labeled probe, the present invention does not specifically limit the dosage form and administration method of the pharmaceutical composition, and the dosage form and administration method well known to those skilled in the art can be used.

[0240] The present invention also provides a lyophilization kit, comprising a precursor compound and a pharmaceutically acceptable excipient, wherein the precursor compound comprises the PSMA-targeted small molecule compound described in the above technical solution and one or more of the pharmaceutically acceptable forms of the PSMA-targeted small molecule compound described in the above technical solution.

[0241] The present invention provides the use of the PSMA-targeted small molecule compound described in the above technical solution, the radionuclide-labeled probe described in the above technical solution and / or the pharmaceutically acceptable form of existence described in the above technical solution, the pharmaceutical composition described in the above technical solution or the freeze-dried kit described in the above technical solution in the preparation of therapeutic agents, therapeutic drugs, diagnostic agents or diagnostic drugs for diseases mediated by PSMA proteins. In the present invention, the PSMA protein-mediated disease preferably includes tumors and / or metastases thereof; the tumor preferably includes one or more of prostate cancer, breast cancer, ovarian cancer, liver cancer, lung cancer, colorectal cancer, bone tissue sarcoma, connective tissue sarcoma, renal cell carcinoma, gastric cancer, pancreatic cancer, nasopharyngeal cancer, head and neck cancer, neuroendocrine tumor, skin melanoma and bone metastasis, more preferably prostate cancer. In the present invention, the diagnostic methods of the diagnostic reagents and diagnostic drugs independently preferably include one or more of single photon emission computed tomography (SPECT), positron emission tomography (PET) and magnetic resonance imaging (MRI); the treatment methods of the therapeutic reagents and therapeutic drugs independently preferably include radionuclide targeted therapy and / or chemotherapy.

[0242] The PSMA-targeted small molecule compound, the PSMA-targeted small molecule compound and its pharmaceutically acceptable form, the radionuclide-labeled probe and its pharmaceutically acceptable form provided by the present invention all have excellent in vivo biological properties, have excellent lesion uptake in lesions with high expression of PSMA protein, and have a long retention time; and are quickly cleared in non-target organs such as normal tissues with a low signal background, and therefore have the characteristics of a high target / non-target ratio, are particularly suitable for radionuclide therapy and imaging of tumors, can improve the diagnostic and therapeutic effects of PSMA-targeted radionuclide, and have the potential for clinical promotion and application.

[0243] In order to further illustrate the present invention, the technical solution of the present invention is described in detail below in conjunction with the accompanying drawings and embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0244] Example 1

[0245] Synthesis of DNB8P1

[0246]

[0247] The synthesis process is as follows: 3.18 μmol of precursor NB8P1, 7.95 μmol of hydroxysuccinimide-tetraazacyclododecanetetraacetic acid (DOTA-NHS) and 15.91 μmol of N,N-diisopropylethylamine (DIPEA) were dissolved in 500 μL of dimethyl sulfoxide (DMSO), stirred at room temperature for 12 h, separated and purified by semi-preparative HPLC, and the target product peak was collected and freeze-dried to obtain DNB8P1.

[0248] Semi-preparative HPLC separation and purification conditions: reverse phase C18 semi-preparative column (10mm×250mm); mobile phase A: water + 0.1v / v% trifluoroacetic acid (TFA); mobile phase B: acetonitrile + 0.1v / v% TFA; gradient elution conditions: 0-10min, the volume fraction of mobile phase B increased from 5% to 50%, 10-15min: the mobile phase ratio remained unchanged, 15-17min: the volume fraction of mobile phase B decreased to 5% and maintained until 30min. The mobile phase flow rate was 3mL / min.

[0249] HPLC purity analysis conditions: use a reverse phase C18 analytical column (4.6×250 mm); mobile phase A: water + 0.1 v / v% TFA; mobile phase B: acetonitrile + 0.1 v / v% TFA; gradient elution program: 0-10 min, the volume fraction of mobile phase B increases from 5% to 50%, 10-15 min: the volume fraction of mobile phase B is 50%; 15-17 min: the volume fraction of mobile phase B decreases from 50% to 5%, 17-30 min, the volume fraction of mobile phase B is 5%; the mobile phase flow rate is 1 mL / min; the radioactive signal is detected by a radioactivity detector.

[0250] Embodiments 2 to 17

[0251] According to the preparation method of DNB8P1, by replacing the structures of n, R1, R2 and R3 in compound II, the corresponding PSMA targeted small molecule compounds DNA8P1, DNC8P1, DND8P1, DNA6P1, DNB6P1, DCA8P1, DCB8P1, DNB8P2, DCB8P2, NNA8P1, NCB8P1, NNB8P1, DGNB8P1, DSNB8P1, DGNE8P1 and DGNF8P1 were obtained respectively. n, R1, R2 and R3 are shown in Table 17.

[0252] The mass spectra of the PSMA-targeted small molecule compounds prepared in Examples 1 to 17 are shown in Figures 1 to 17 It can be seen that the target PSMA-targeted small molecule compound was successfully prepared by the present invention.

[0253] The HPLC purity analysis chart of the PSMA targeted small molecule compounds prepared in Examples 1 to 17 is shown in Figures 18 to 25 It can be seen that the HPLC purity of each PSMA-targeted small molecule compound is greater than 95%.

[0254] Table 17 n, R1, R2 and R3 in the PSMA-targeted small molecule compounds prepared in Examples 1 to 17

[0255]

[0256] Embodiment 18

[0257] 68 Ga-labeled radionuclide probe [ 68 Synthesis of Ga]Ga-DNB8P1

[0258]

[0259] The labeling method is as follows: 50 μg of the compound DNB8P1 prepared in Example 1 is dissolved in 500 μL of 0.3 M acetic acid-sodium acetate buffer solution, and then added to 2 mL of about 185-555 MBq of the compound eluted from the germanium gallium generator. 68 Place the GaCl3 hydrochloric acid solution in a vial, shake at 95°C for 20 minutes, then cool to room temperature, dilute with saline or water for injection, and filter through a sterile filter membrane to obtain the radionuclide. 68 Ga-labeled [ 68 Ga]Ga-DNB8P1 solution.

[0260] The radionuclide labeled probe was subjected to radio-HPLC purity analysis and identification, using a reverse phase C18 analytical column (4.6×250 mm). Mobile phase A: water + 0.1 v / v% trifluoroacetic acid (TFA); mobile phase B: acetonitrile + 0.1 v / v% TFA. Gradient elution program: 0-10 min, the volume fraction of mobile phase B increased from 5% to 50%, 10-15 min: the volume fraction of mobile phase B was 50%; 15-17 min: the volume fraction of mobile phase B decreased from 50% to 5%, 17-30 min, the volume fraction of mobile phase B was 5%; the mobile phase flow rate was 1 mL / min; the radioactive signal was detected by a radioactive detector.

[0261] Examples 19 to 31

[0262] according to[ 68 The PSMA-targeted small molecule compound DNB8P1 was replaced with DNA8P1, DNC8P1, DND8P1, DCA8P1, DCB8P1, DNA6P1, DNB6P1, DNB8P2, DCB8P2, DGNB8P1, DSNB8P1, DGNE8P1 and DGNF8P1 to obtain radionuclide labeled probes [ 68 Ga]Ga-DNA8P1, [ 68 Ga]Ga-DNC8P1, [ 68 Ga]Ga-DND8P1, [ 68 Ga]Ga-DCA8P1, [ 68 Ga]Ga-DCB8P1, [ 68 Ga]Ga-DNA6P1, [68 Ga]Ga-DNB6P1, [ 68 Ga]Ga-DNB8P2, [ 68 Ga]Ga-DCB8P2, [ 68 Ga]Ga-DGNB8P1, [ 68 Ga]Ga-DSNB8P1, [ 68 Ga]Ga-DGNE8P1 and [ 68 The radio-HPLC results of the nuclide labeled probes prepared in Examples 18 to 31 are shown in Figures 26 to 32 It can be seen that the synthesized 68 The radiochemical purity of Ga-labeled probes was greater than 95%.

[0263] Embodiment 32

[0264] 18 F-labeled radionuclide-labeled probe [ 18 Synthesis of F]AlF-NNB8P1

[0265]

[0266] 50 μg of the labeled precursor NNB8P1 dissolved in 20 μL of dimethyl sulfoxide was mixed with 100 μL of 0.1 M acetic acid-sodium acetate buffer solution (pH = 4), 200 μL of acetonitrile and 3 μL of 2 mM AlCl3 solution (prepared with 0.1 M acetic acid-sodium acetate buffer solution at pH = 4), and 100 μL of about 740 MBq of radioactive nuclides produced by the accelerator was added. 18 F - The solution was shaken at 100℃ for 20 minutes. Take a Sep-Pak C18 separation column and activate it by eluting it with 10mL of anhydrous ethanol and 10mL of water. Dilute the labeled solution with 8mL of pure water and load it onto the separation column. Rinse the C18 column with 8mL of pure water to remove the unreacted 18 F - Ions. Elute with 1.5 mL of anhydrous ethanol solution, remove the organic solvent by nitrogen blowing, dilute with physiological saline or water for injection, and filter through a sterile filter membrane to obtain the radionuclide. 18 F-labeled complexes [ 18 The [F]AlF-NNB8P1 solution was subjected to radio-HPLC analysis and identification according to the conditions of Example 18.

[0267] Embodiments 33-34

[0268] according to[ 18F]AlF-NNB8P1 was prepared by replacing NNB8P1 with NNA8P1 and NCB8P1, respectively, to obtain probes [ 18 F]AlF-NNA8P1 and [ 18 F]AlF-NCB8P1.

[0269] The probes prepared in Examples 32 to 34 were sampled and analyzed by HPLC. The HPLC results are shown in Fig.33 It can be seen that the present invention prepares 18 The radiochemical purity of the F-labeled probes was greater than 95%.

[0270] Embodiment 35

[0271] 177 Lu-labeled radionuclide-labeled probe [ 177 Synthesis of Lu]Lu-DGNB8P1

[0272]

[0273] 370MBq 177 LuCl3 solution was added to a centrifuge tube containing 0.2 mL of acetic acid-sodium acetate solution (0.5 M, pH = 4.0) containing DGNB8P1 (50 μg), placed at 95°C for 25 min, cooled to room temperature, diluted with physiological saline or water for injection, and sterile filtered to obtain the radionuclide labeled probe [ 177 The Lu]Lu-DGNB8P1 solution was subjected to radio-HPLC analysis and identification according to the conditions of Example 18.

[0274] Embodiments 36 to 38

[0275] according to[ 177 The probes [Lu]Lu-DGNB8P1 were prepared by replacing DGNB8P1 with DSNB8P1, DGNE8P1 and DGNF8P1, respectively. 177 Lu]Lu-DSNB8P1, [ 177 Lu]Lu-DGNE8P1 and [ 177 Lu]Lu-DGNF8P1.

[0276] According to the conditions of Example 18, samples of each radionuclide labeled probe were sampled and analyzed by radio-HPLC. The radio-HPLC results of each probe prepared in Examples 35 to 38 are shown in Fig.34 It can be seen that the present invention prepares 177 The radiochemical purity of the Lu-labeled nuclide-labeled probes was greater than 95%.

[0277] Embodiment 39

[0278] 44 Sc-labeled radionuclide-labeled probe [ 44 Synthesis of Sc]Sc-DGNB8P1

[0279]

[0280] 50 μg of the compound DGNB8P1 prepared in Example 2 was mixed with about 5.55 MBq of accelerator-prepared radionuclides. 44 Sc - The solution (2.6 mL, adjusted to pH = 4 with ammonium acetate) was mixed in a vial, heated at 100°C for 20 minutes, and then reduced to about 300 μL by nitrogen blowing. After cooling to room temperature, it was diluted with physiological saline or water for injection and filtered through a sterile filter membrane to obtain the radionuclide. 44 Sc-labeled [ 44 The Sc]Sc-DGNB8P1 solution was subjected to radio-HPLC purity analysis and identification according to the conditions of Example 18.

[0281] Embodiment 40

[0282] according to[ 44 Sc]Sc-DGNB8P1 was prepared by replacing DGNB8P1 with DNB8P1 to obtain the probe [ 44 Sc]Sc-DNB8P1.

[0283] According to the conditions of Example 18, the radionuclide-labeled probe was sampled and analyzed and identified by radio-HPLC.

[0284] Fig.35 For the probe 44 Sc]Sc-DNB8P1 and [ 44 Sc]Sc-DGNB8P1 HPLC chart, it can be seen that the prepared 44 The radiochemical purity of the Sc-labeled nuclide-labeled probes was greater than 95%.

[0285] Embodiment 41

[0286] 131 I-labeled radionuclide probe 131 Synthesis of IPB-NB8P1

[0287]

[0288] Dissolve 500 μg of NB8P1 in 200 μL of anhydrous DMSO. 131After mixing IPBA-NHS and NB8P1 solution, 5-10 μL N,N-diisopropylethylamine was added. The reaction was carried out at room temperature for 2 hours. After the reaction was completed, the product was purified by radio-HPLC and the radiochemical purity was further analyzed.

[0289] The radionuclide labeled probe was subjected to radio-HPLC purity analysis and identification. A reverse phase C18 analytical column (4.6×250 mm) was used. Mobile phase A: water + 0.1 v / v% trifluoroacetic acid (TFA); mobile phase B: acetonitrile + 0.1 v / % TFA; gradient elution program: 0-20 min, the volume fraction of mobile phase B increased from 5% to 95%; 20-22 min: the volume fraction of mobile phase B was 95%; 22-25 min: the volume fraction of mobile phase B decreased from 95% to 5%; the mobile phase flow rate was 1 mL / min. The radioactive signal was detected by a radioactive detector. Fig.36 For probe 131 The HPLC chart of IPB-NB8P1 shows that the radioactivity 131 The radiochemical purity of IPB-NB8P1 was greater than 95%.

[0290] Test Example 1

[0291] Evaluation of the stability and lipid-water distribution properties of radionuclide labeled probes

[0292] 1. Injection stability experiment: Place the radionuclide-labeled probe in physiological saline or in vitro serum at room temperature for different periods of time, and take samples for analysis by HPLC. 68 Ga, 18 F. 177 Lu 44 The HPLC analysis conditions of the Sc nuclide-labeled probe are the same as those of the corresponding probes in Example 18, Example 32, Example 35, and Example 39, respectively.

[0293] Figures 37 to 51 The probes are 68 Ga]Ga-DNA8P1, [ 68 Ga]Ga-DNB8P1, [ 68 Ga]Ga-DNA6P1, [ 68 Ga]Ga-DNB6P1, [ 68 Ga]Ga-DNC8P1, [ 68 Ga]Ga-DND8P1, [ 68 Ga]Ga-DCA8P1, [ 68 Ga]Ga-DCB8P1, [ 18 F]AlF-NNA8P1, [ 18 F]AlF-NNB8P1, [18 F]AlF-NCB8P1, [ 177 Lu]Lu-DGNB8P1, [ 177 Lu]Lu-DGNF8P1, [ 44 Sc]Sc-DNB8P1 and [ 44 HPLC chart of the stability of Sc]Sc-DGNB8P1 in physiological saline or in vitro serum shows that at the tested time points, 68 Ga, 18 F. 177 Lu 44 The radiochemical purity of the Sc-labeled nuclide-labeled probes was greater than 95%, indicating that they were stable in the specified medium.

[0294] 2. Determination of lipid-water distribution coefficient (logP):

[0295] 100 μL of radionuclide labeled probe solution (3.7 MBq / mL) diluted with PBS (pH = 7.4) was added to a centrifuge tube containing a mixture of 0.9 mL PBS (pH = 7.4) and 1 mL n-octanol. After vortexing for 3 minutes, the mixture was centrifuged at 10000 r / min for 3 minutes. 100 μL of liquid was taken from each of the aqueous phase and n-octanol phase and the radioactivity count was determined by a γ-counter. The experiment was repeated three times and the average value was taken. The calculation formula of logP is shown in formula (1):

[0296] P=I 有机相 / I 水相 Formula (1);

[0297] Among them, I 有机相 represents the radioactivity count measured in the organic phase, I 水相 Represents the radioactive count measured in the aqueous phase. The lipid-water distribution coefficient results of each radiolabeled probe are shown in Table 18.

[0298] Table 18 Lipid-water distribution coefficient of radionuclide labeled probe

[0299]

[0300]

[0301] From Table 18, we can see that 68 Ga, 18 F. 117 Lu 44 Sc-labeled radionuclide-labeled probes are all hydrophilic and have good water solubility. This property helps to reduce the retention of probes in organs such as the liver, gallbladder, and intestines, and avoid nonspecific uptake of radioactivity in normal tissues.

[0302] Test Example 2

[0303] Radionuclide labeled probe receptor affinity and cellular uptake test

[0304] 1. Test of the receptor affinity of the probe in PSMA-positive cells PC3-PIP

[0305] Affinity test method: 48 hours in advance, inoculate about 2×10 5 PC3-PIP cells. At the beginning of the experiment, the old culture medium was first removed and the cells were washed with 500 μL cold PBS (pH = 7.4, containing 0.2% BSA) per well. 100 μL of unlabeled PSMA-617 (10 μg / well, prepared in serum-free RPMI-1640 culture medium) was added to each well of one of the 24-well plates, and an equal volume of culture medium was added to the other plate. 100 μL of different concentrations (0.156-400 nM) of 68 Ga labeled radioligand, each concentration was repeated 3 wells. After incubation at 37 ° C for 0.5 h, the cells were washed twice with cold PBS containing 0.2% BSA (500 μL / well). Then the cells were lysed with 1M NaOH (500 μL / well), and the radioactivity of the lysate was measured with a γ counter.

[0306] Figures 52-55 The probes are 68 Ga]Ga-DNA8P1 and [ 68 Ga]Ga-DNB8P1, [ 68 Ga]Ga-DNC8P1 and [ 68 Ga]Ga-DND8P1, [ 68 Ga]Ga-DNA6P1 and [ 68 Ga]Ga-DNB6P1, [ 68 Ga]Ga-DCA8P1 and [ 68 The K of Ga]Ga-DCB8P1 in PSMA-positive cells PC3-PIP d Test curve. Fig.56 for[ 18 F]AlF-NNA8P1, [ 18 F]AlF-NNB8P1 and [ 18 F]AlF-NCB8P1 K in PSMA-positive cells PC3-PIP d Test curve. The K of the above radionuclide labeled probes d The values ​​are shown in Table 19, and all the probes tested have good affinity for PSMA.

[0307] Table 19 K of radionuclide labeled probe d value

[0308] Probe <![CDATA[K d (nM)]]> Probe <![CDATA[K d (nM)]]> Probe <![CDATA[K d (nM)]]> <![CDATA[[ 68 Ga]Ga-DNA8P1]]> 97.61 <![CDATA[[ 68 Ga]Ga-DNA6P1]]> 158.10 <![CDATA[[ 18 F]AlF-NNA8P1]]> 97.54 <![CDATA[[ 68 Ga]Ga-DNB8P1]]> 112.40 <![CDATA[[ 68 Ga]Ga-DNB6P1]]> 68.62 <![CDATA[[ 18 F]AlF-NNB8P1]]> 175.5 <![CDATA[[ 68 Ga]Ga-DNC8P1]]> 37.36 <![CDATA[[ 68 Ga]Ga-DCA8P1]]> 133.40 <![CDATA[[ 18 F]AlF-NCB8P1]]> 114.6 <![CDATA[[ 68 Ga]Ga-DND8P1]]> 155.20 <![CDATA[[ 68 Ga]Ga-DCB8P1]]> 89.77

[0309] 2. Radionuclide-labeled probe cell uptake and inhibition experiments

[0310] PC3-PIP cells with high PSMA expression and PC3 cells with negative expression were plated in a 24-well plate containing culture medium (containing fetal bovine serum and double antibody) (cell counting plate counted the number of cells, about 200,000 / well) and cultured for 48 hours. At the beginning of the uptake experiment, the original culture medium was aspirated and the cells were washed with PBS (500 μL, pH=7.4, containing 0.2% BSA); an equal amount of the radionuclide labeled probe to be tested diluted with culture medium without fetal bovine serum and double antibody was added to each well, and incubated at different time points at 37°C; after the incubation at each time point was completed, the culture medium was aspirated, and sodium hydroxide (NaOH) solution (500 μL, 1M) was added to each well to lyse the cells. After 5 minutes, the lysed cells were placed in a disposable centrifuge tube to measure the radioactivity count; the count was divided by the total amount of added radioactivity to obtain the cell uptake percentage.

[0311] In order to investigate the PSMA targeting specificity of the radionuclide labeled probe, an inhibition group was set up at the same time, that is, an appropriate amount of inhibitor PSMA-617 was added to each well of cells before adding the radionuclide labeled probe, and the cells were incubated at 37°C for a certain period of time ( 18 The F-labeled probe was co-incubated for 1 h; 177 Lu probe was co-incubated for 2h and 4h); after the incubation was completed, the radioactive medium was aspirated, and sodium hydroxide solution (500μL, 1M) was added to each well to lyse the cells. After 5min, the lysed cells were placed in a disposable centrifuge tube to measure the radioactivity count; the count was divided by the total amount of added radioactivity to obtain the cell uptake percentage.

[0312] Fig.57 For radionuclide labeled probes [ 18 F]AlF-NNA8P1, [ 18 F]AlF-NNB8P1 and [ 18 The results of cell uptake and cell inhibition of F]AlF-NCB8P1 in PSMA-positive cells PC3-PIP at different times (60 min time point) were compared with the results of cell uptake of the probe in PSMA-negative cells PC3 at 60 min. It can be seen that, taking the results at 60 min time point as an example, each radionuclide-labeled probe was significantly taken up in PSMA-positive cells, while the uptake in negative cells was significantly reduced. The uptake of the probe in PSMA-positive cells can be inhibited by PSMA-617, indicating that the [ 18 F]AlF-NNA8P1, [ 18 F]AlF-NNB8P1 and [ 18F]AlF-NCB8P1 probe is specific for targeting PSMA protein.

[0313] Fig.58 For radionuclide labeled probes [ 177 Lu]Lu-DGNB8P1 and [ 177 The cell uptake results and cell inhibition results of Lu]Lu-DSNB8P1 in PSMA positive cells PC3-PIP at different times (2h and 4h time points) were compared with the cell uptake results of radionuclide labeled probe in negative cells PC3 at 2h and 4h. It can be seen that, taking the results at 4h as an example, [ 177 Lu]Lu-DGNB8P1 and [ 177 Lu]Lu-DSNB8P1 was significantly taken up in PSMA-positive cells, while the uptake in PSMA-negative cells was significantly reduced. The uptake of radionuclide-labeled probes in PSMA-positive cells was inhibited by PSMA-617, indicating that the [ 177 Lu]Lu-DGNB8P1 and [ 177 The Lu]Lu-DSNB8P1 radionuclide-labeled probe is specific for targeting PSMA protein.

[0314] Test Example 3

[0315] PET imaging experiment with radionuclide labeled probes

[0316] The radiochemical purity of the prepared samples is greater than 95%. 68 Ga or 18 After the F-labeled probe was diluted with saline, 0.2 mL (about 7.4 MBq) of radionuclide labeled probe injection solution was injected through the tail vein of PC3-PIP model mice. MicroPET imaging was performed at different time points, and the region of interest (ROI) was outlined on the image, and the distribution value of the radionuclide labeled probe was obtained by calculation.

[0317] Figures 59 to 72 The probes are 68 Ga]Ga-DNA8P1, [ 68 Ga]Ga-DNB8P1, [ 68 Ga]Ga-DNC8P1, [ 68 Ga]Ga-DND8P1, [ 68 Ga]Ga-DNA6P1, [ 68 Ga]Ga-DNB6P1, [ 68 Ga]Ga-DCA8P1, [ 68 Ga]Ga-DCB8P1, [ 68 Ga]Ga-DNB8P2, [ 68Ga]Ga-DCB8P2, [ 68 Ga]Ga-DGNB8P1, [ 68 Ga]Ga-DSNB8P1, [ 68 Ga]Ga-DGNE8P1 and [ 68 PET imaging of Ga]Ga-DGNF8P1 (A) and uptake values ​​of major tissues and target / non-target ratios (B). It can be seen that the radionuclide-labeled probe has high uptake at the tumor site. There is a slight radioactive signal at the early time point in the kidney, but it quickly disappears over time. The background value of the radionuclide-labeled probe in other non-target organs is very low, and the absolute uptake of the tumor and the target / non-target ratio increase significantly over time.

[0318] Fig.73 For different contents of competitive binders [DNB8P1, nat Ga-DNB8P1 (non-radioactive gallium), PSMA-617] 68 Comparison of PET imaging results (A) and uptake values ​​of major tissues (B) of Ga]Ga-DNB8P1. nat When Ga-DNB8P1 or PSMA-617, the probe [ 68 The uptake of Ga]Ga-DNB8P1 in the tumor site was significantly weakened. However, the presence of a certain dose of DNB8P1 did not affect the probe [ 68 Ga]Ga-DNB8P1 had a significant impact on the PET imaging quality and tumor uptake.

[0319] Fig.74 For radionuclide labeled probes [ 68 Ga]Ga-DNB8P1 and classical probes [ 68 Comparison of PET imaging (A), uptake values ​​of major tissues (B), and target / non-target ratio (C) of Ga]Ga-PSMA-617 (self-made, prepared according to the process of Example 18). 68 Ga]Ga-PSMA-617, [ 68 Ga]Ga-DNB8P1 has significant improvements in PET imaging quality, absolute tumor uptake, and target / non-target ratio.

[0320] Figures 75-77 The radionuclide labeled probes are 18 F]AlF-NNA8P1, [ 18 F]AlF-NNB8P1 and [ 18 F] PET imaging of AlF-NCB8P1 (A) and uptake values ​​of major tissues (B), Figures 75-77The 1h inhibition group used PSMA-617 as an inhibitor. It can be seen that the radionuclide-labeled probe has high uptake at the tumor site. The radioactive signal at the kidney position is quickly cleared over time. The background value of the radionuclide-labeled probe in other non-target organs is very low, and the absolute uptake of the tumor is well maintained over time, which helps to achieve better quality of results.

[0321] Take 0.2 mL (0.37 MBq) of the radiochemical purity of more than 95% prepared in Example [ 44 Sc]Sc-DGNB8P1 injection was injected into PC3-PIP model mice via the tail vein. MicroPET imaging was performed at different time points, and the region of interest (ROI) was outlined on the image, and the probe distribution value was obtained by calculation. Fig.78 For radionuclide labeled probes [ 44 PET imaging of Sc]Sc-DGNB8P1 (A), uptake values ​​of major tissues, and target / non-target ratio comparison (B). The radionuclide-labeled probe was significantly uptaken in the tumor site at a short time point of 15 minutes after injection. The probe was mainly metabolized via the kidney pathway. Over time, the radionuclide-labeled probe was significantly metabolized in non-target organs, while the uptake in the tumor was able to remain stable, showing an improved target-non-target ratio.

[0322] Test Example 4

[0323] SPECT imaging experiment with radionuclide labeled probes

[0324] Tumor-bearing mice were injected with 37MBq via tail vein 177 Lu-labeled radionuclide-labeled probe ([ 177 Lu]Lu-DGNB8P1, [ 177 Lu]Lu-DSNB8P1, [ 177 Lu]Lu-DGNF8P1 and [ 177 Lu]Lu-PSMA-617 (one of the gold standards, homemade, prepared according to the process of Example 35), static SPECT scanning imaging was performed at different time points after injection. After the imaging was completed, the image was reconstructed and the region of interest (ROI) in the mouse image was outlined to obtain the radioactive count value, and the target / non-target ratio of the radionuclide labeled probe distribution was calculated.

[0325] Radionuclide labeled probes 177 Lu]Lu-DGNB8P1, [ 177 Lu]Lu-DSNB8P1, [ 177 The SPECT imaging results of Lu]Lu-DGNF8P1 are as follows Figures 79-81 As shown. Control group [ 177The SPECT imaging results of Lu]Lu-PSMA-617 are as follows Fig.82 As shown. It can be seen that within the monitoring time range, [ 177 Lu]Lu-DGNB8P1, [ 177 Lu]Lu-DSNB8P1, [ 177 The enrichment of Lu-DGNF8P1 nuclide-labeled probe in the tumor site was higher than that in the control group [ 177 The results show that the radionuclide labeled probe provided by the present invention has good tumor uptake effect and has the potential for PSMA radionuclide targeted therapy.

[0326] Fig.83 In the presence of 20 μg of competitive binding agent DGNB8P1, the radionuclide-labeled probe [ 177 SPECT imaging results and target / non-target ratio of Lu]Lu-DGNB8P1. It can be seen that when there is 20μg DGNB8P1 in the injection solution, the radionuclide labeled probe [ 177 The SPECT imaging quality of Lu]Lu-DGNB8P1 was not affected, and it still had good tumor uptake and a high tumor / kidney ratio.

[0327] Test Example 5

[0328] Biodistribution experiments with radionuclide-labeled probes

[0329] Tumor-bearing mice were injected with the radionuclide-labeled probe prepared in Example ([ 18 F]AlF-NNA8P1, [ 18 F]AlF-NCB8P1, [ 177 Lu]Lu-DGNB8P1 or [ 44 Sc]Sc-DGNB8P1) injection. Mice were killed at different time points after injection, and tumor and other organ tissue samples were obtained by dissection, weighed, and radioactivity counts were measured using a γ counter. The results were expressed as the percentage of uptake dose per gram of tissue or organ (%ID / g).

[0330] Figures 84-85 The radionuclide labeled probes are 18 F]AlF-NNA8P1 and [ 18 The biodistribution results of F]AlF-NCB8P1 in tumor-bearing mice. It can be seen that both radionuclide-labeled probes have high uptake in the tumor site, slight retention in the kidney, and can be quickly cleared. In normal tissues, both radionuclide-labeled probes show low background signals, which is very conducive to PSMA-targeted PET imaging.

[0331] Fig.86For radionuclide labeled probes [ 177 The biodistribution results of Lu]Lu-DGNB8P1 in tumor-bearing mice. It can be seen that 4h after injection, [ 177 The tumor uptake of Lu]Lu-DGNB8P1 was greater than 30% ID / g. 24h after injection, the tumor uptake value was still higher than 20% ID / g and maintained at a high level at 96h. 177 Lu]Lu-DGNB8P1 has good tumor uptake and prolonged retention time, which is very beneficial for PSMA-targeted radionuclide therapy.

[0332] Fig.87 For radionuclide labeled probes [ 44 Sc]Sc-DGNB8P1 uptake values ​​of various organs in tumor-bearing mice at different time points. The radionuclide-labeled probe is rapidly enriched in the tumor and remains for a long time. The uptake value in most other organs is low and decays over time. The probe is mainly metabolized through the kidney. The inhibition group injected with 100 μg of competitive binder PSMA-617 can significantly inhibit [ 44 Tumor uptake of Sc]Sc-DGNB8P1.

[0333] Test Example 6

[0334] PSMA-targeted radionuclide therapy experiment using radionuclide labeled probes

[0335] The tumor-bearing mice were divided into experimental groups ([ 177 Lu]Lu-DGNB8P1 group), [ 177 Lu]Lu-PSMA-617 control group and saline group. Each mouse in the experimental group was injected with different doses of [ 177 Lu]Lu-DGNB8P1 (37MBq, 18.5MBq and 9.25MBq, respectively); [ 177 Each mouse in the Lu-PSMA-617 control group was injected with 18.5 MBq [ 177 The same volume of saline was injected into the tail vein of each mouse in the Lu]Lu-PSMA-617; saline group, and the tumor size and body weight were monitored every day. Fig.88 As shown, compared with the saline group, the tumor volume in the experimental group decreased significantly over time. 177 Lu]Lu-DGNB8P1 have obvious tumor treatment effects. 177 In the Lu]Lu-PSMA-617 control group, tumor growth was suppressed in the early stage of treatment, but rebounded significantly in the later stage of treatment. It is worth mentioning that the 18.5MBq and 9.25MBq doses of [ 177The inhibitory effect of Lu]Lu-DGNB8P1 on tumors was significantly better than that of [ 177 Lu]Lu-PSMA-617, indicating that the present invention provides [ 177 The Lu]Lu-DGNB8P1 probe has good therapeutic application prospects.

[0336] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A PSMA-targeted small molecule compound having a structure shown in Formula I: In the formula I, n is an integer from 0 to 5; R1 includes any of the following structures: R2 may be present or absent, and when present, R2 includes any one of the following structures: In said R2, a, b and t are independently integers between 0 and 5; R3 is a group to be labeled, including any one of the following chelating structures:

2. The method for preparing the PSMA-targeted small molecule compound according to claim 1, comprising method 1, method 2 or method 3; The method 1 comprises the following steps: The PSMA-targeted small molecule compound is obtained by performing a substitution reaction with the compound 1 using an R3 active compound having an activated ester group or an acid anhydride group to form an amide bond; the R3 active compound having an activated ester group or an acid anhydride group includes any one of the following structures: The method 2 comprises the following steps: using an R3 active compound with an isothiocyanate group to perform a substitution reaction with compound 1 to form a thiourea bond, thereby obtaining the PSMA-targeted small molecule compound; the R3 active compound with an isothiocyanate group comprises any one of the following structures: The method 3 comprises the following steps: using an R3 chelating agent with a carboxyl structure to perform an amidation reaction with compound 1 under condensation reagent conditions to generate an amide bond, and then removing the protecting group to obtain the PSMA-targeted small molecule compound; the R3 chelating agent with a carboxyl structure comprises any one of the following structures, wherein A is a carboxyl protecting group: R1, R2 and n in the compound 1 are the same as R1, R2 and n in the formula I.

3. A radionuclide labeled probe, characterized in that: It has a structure shown in formula II or formula III: Wherein, the definitions of R1, R2 and n are the same as those of R1, R2 and n in Formula I; R4 is a nuclide labeling group obtained by a coordination reaction between the R3 to-be-labeled group in the PSMA-targeted small molecule compound of claim 1 and the nuclide; and R5 includes any one of the following structures: Wherein, y and z are independently integers between 0 and 5; X is carbon, hydrogen or nitrogen; R6 is 18 F. 123 I. 124 I. 125 I. 131 I or 211 At.

4. The nuclide-labeled probe according to claim 3, characterized in that: The nuclides include 18 F. 43 Sc, 44 Sc, 47 Sc, 51 Cr, 55 Co. 57 Co. 62 Cu, 64 Cu, 67 Cu, 67 Ga, 68 Ga, 72 As, 72 Se, 89 Zr, 86 Y. 89 Sr. 86 Y. 90 Y. 97 Such as 99m Tc, 105 Rh, 101m Rh, 109 Pd, 111 In, 119 Sb, 128 8. 139 La, 140 La, 142 Pr, 149 Pm, 149 Tb, 151 Tb, 151 Eu, 153 Eu, 169 Eu, 153 Sm, 152 Gd, 153 Gd, 157 Gd, 159 Gd, 161 Tb, 165 Dy, 166 Ho, 169 2. 175 Yb, 177 Lu, 186 Re, 188 Re, 197 Hg, 198 Au, 201 Tl, 203 Pb, 211 At 212 Pb, 212 Bi, 213 Bi, 223 Ra, 227 Th and 225 At least one of Ac.

5. A method for preparing the radionuclide-labeled probe according to claim 3 or 4, The method for preparing the nuclide-labeled probe having the structure shown in Formula II comprises the following steps: Conducting a coordination reaction between the PSMA-targeted small molecule compound of claim 1 and a nuclide to obtain a nuclide-labeled probe having a structure shown in Formula II; The method for preparing the nuclide-labeled probe having the structure shown in Formula III comprises the following steps: subjecting Compound 1 to an amidation reaction with an active compound to obtain a nuclide-labeled probe having the structure shown in Formula III; the active compound comprises Compound 2 or Compound 3; 6. A pharmaceutically acceptable form, characterized in that: The invention relates to a pharmaceutically acceptable form of the PSMA-targeted small molecule compound of claim 1 and / or a pharmaceutically acceptable form of the radionuclide-labeled probe of claim 3 or 4, wherein the pharmaceutically acceptable form comprises one or more of a salt, a stereoisomer, a racemate, a hydrate and a solvate.

7. A pharmaceutical composition, characterized in that The invention comprises an active component and a pharmaceutically acceptable excipient; the active component comprises one or more of the PSMA-targeted small molecule compound according to claim 1, the radionuclide-labeled probe according to claim 3 or 4, and the pharmaceutically acceptable form according to claim 6.

8. A freeze-dried kit, characterized in that: The method comprises a precursor compound and a pharmaceutically acceptable excipient, wherein the precursor compound comprises one or more pharmaceutically acceptable forms of the PSMA-targeted small molecule compound according to claim 1 and the PSMA-targeted small molecule compound according to claim 6.

9. Use of the PSMA-targeted small molecule compound of claim 1, the radionuclide-labeled probe of claim 3 or 4, the pharmaceutically acceptable form of the radionuclide-labeled probe of claim 6, the pharmaceutical composition of claim 7 or the lyophilized kit of claim 8 in the preparation of therapeutic agents, therapeutic drugs, diagnostic agents or diagnostic drugs for diseases mediated by PSMA protein.

10. The use according to claim 9, characterized in that: The PSMA protein-mediated diseases include tumors and / or their metastases; The tumor includes one or more of prostate cancer, breast cancer, ovarian cancer, liver cancer, lung cancer, colorectal cancer, bone tissue sarcoma, connective tissue sarcoma, renal cell carcinoma, gastric cancer, pancreatic cancer, nasopharyngeal carcinoma, head and neck cancer, neuroendocrine tumors, skin melanoma and bone metastasis.

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