Peptide urea derivatives, pharmaceutical compositions containing same and uses thereof

Abstract

Disclosed herein are peptide urea derivatives, pharmaceutical compositions thereof and uses thereof. The derivatives are represented by formula I, in which R is a group containing a radioactive metal ion or an optically imageable group. The derivatives can be used for preoperative imaging diagnosis and grading of PSMA-positive prostate cancer, and can also be used for the treatment of various types and stages of prostate cancer, realizing the integration of diagnosis and treatment, and are expected to be widely used. JPEG2025505467000166.jpg32157

Description

[Technical field]

[0001] This application claims priority to Chinese patent application No. 202210116230.9, filed on January 30, 2022. This application cites the above-mentioned Chinese patent application in full. The present invention relates to peptide urea derivatives, pharmaceutical compositions containing same and uses thereof. [Background technology]

[0002] According to the Global Cancer Statistics 2018 report published in the online edition of the Journal of Cancer in Clinicians, the official journal of the American Cancer Society, the incidence and mortality rates of 36 cancers in 185 countries were evaluated and found that prostate cancer is the second most prevalent cancer in men after lung cancer. According to the US Cancer Statistics 2018 report, the incidence rate of prostate cancer in American men is predicted to be the first, accounting for about 19% of the tumor incidence rate. According to the 2014 national cancer statistics published by the National Cancer Center of China, the incidence rate of prostate cancer in Chinese men is 3.25%, the sixth highest, but has been gradually increasing in recent years. Therefore, prostate cancer is a type of cancer with a high incidence rate both worldwide and in China.

[0003] Early imaging diagnosis and treatment of prostate cancer has become an urgent problem that needs to be solved not only in China but also around the world. Prostate cancer develops in the tissues around the prostate, and as it grows, it gradually metastasizes to other important organs such as the lungs and bones. There are no obvious symptoms in the early stages, but as prostate cancer grows, it can cause problems such as urethral compression and urinary obstruction, and may even metastasize to the spine and pelvis. Currently, imaging diagnostic methods such as SPECT (single photon emission computed tomography) and PET (positron emission tomography) are used to diagnose prostate cancer, the principle of which is to label PSMA target peptides with radioisotopes that emit gamma rays or positrons, and show the presence and distribution of tumor cells in tomographic and three-dimensional images through prostate cancer-specific target distribution. These imaging diagnostic methods have become widely used in recent years due to the significant improvement in image quality caused by the development of SPECT-CT / MRI and PET-CT / MRI combined with CT or MRI. Radiopharmaceuticals currently used for specific imaging of prostate cancer use PSMA ligands as targeting groups that can bind to PSMA (prostate-specific membrane antigen), a protein specifically expressed in prostate cancer. PSMA is a type II transmembrane glycoprotein, also known as glutamate carboxypeptidase, and is expressed in very low amounts in kidney, small intestine, and brain tissues, with its expression level in tumor tissues being much higher than that in normal tissues. Representative ligands of PSMA are peptide derivatives, such as Glu-urea-Lys (GUL) or Glu-urea-Cys (GUC). Thus, by labeling peptide ligands containing such peptides with radioisotopes, the radiopharmaceuticals produced can be used for imaging prostate cancer by PET or SPECT, or for treating prostate cancer (MEder, et al., Bioconjugate Chem 2012,23:688-697). The radioisotopes used to label peptides are mainly α-, β-, γ-, and positron-emitting radionuclides, where α- and β-emitting radionuclides are used in therapy, and γ- and positron-emitting radionuclides are used in nuclear imaging.There are generally two methods for labeling a ligand with a radioisotope: one is to directly bind the ligand to the radioisotope, and the other is to chelate the radioisotope via a bifunctional chelating agent (BFCA) such as DTPA, DOTA, TETA, HYNIC, N2S2, and MAG3. The direct binding method is mainly used for labeling various non-metallic radioisotopes such as 125I and 131I. The method using a bifunctional chelating agent (BFCA) is mainly used for labeling various metallic radioisotopes, and the type of bifunctional chelating agent (BFCA) can be selected based on the properties of the ligand and the radioisotope.

[0004] Currently, castration surgery, antiandrogen castration, and androgen receptor inhibitors are the mainstream treatment methods for prostate cancer. Although these treatment options are highly effective in the early stages, most patients develop castration-resistant prostate cancer (CRPC) and even metastatic castration-resistant prostate cancer (mCRPC). mCRPC is a disease with limited treatment options and large unmet medical needs, so radiopharmaceuticals targeting PSMA have become a research hotspot in recent years.

[0005] A series of clinical studies have been conducted using radiopharmaceuticals targeting PSMA to treat patients with mCRPC, and although the preliminary clinical results of radiopharmaceuticals such as 177Lu-PSMA-617 and 177Lu-PSMA I&T have been promising, there are some problems, such as the fact that nearly 30% of patients did not respond to this treatment, one possible explanation is that not enough radiopharmaceuticals were delivered to the tumor lesions due to suboptimal pharmacokinetics, and another concern is that radiopharmaceuticals such as 177Lu-PSMA-617 accumulate in organs such as the kidneys and salivary glands for a long time. Therefore, radiopharmaceuticals with high activity, high selectivity, and at the same time better pharmacokinetics targeting PSMA continue to be a hot topic in the field of treatment and diagnosis of mCRPC. Summary of the Invention

[0006] The technical problem that the present invention aims to solve is that the existing peptide urea derivatives have a single structure, and the radiopharmaceutical may be absorbed and retained for a long time by organs other than the target, such as the kidney, and the like, so that the uptake amount in the desired target tumor is not high or the retention time is not long enough. Therefore, the present invention provides a peptide urea derivative, a pharmaceutical composition containing the same, and its use. The derivative can be used for preoperative imaging diagnosis and grading of PSMA-positive prostate cancer, and can also be used for treating various types and stages of prostate cancer, realizing the integration of diagnosis and treatment, and is expected to be widely used. Compared with the reference compound PSMA-617, the compound disclosed in the present invention has better selectivity and pharmacokinetics, and the uptake and retention time of the radiopharmaceutical in the target tumor are both greatly improved.

[0007] The present invention provides a peptide urea derivative represented by Formula I, a pharma- ceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharma- ceutically acceptable salt thereof.

[0008] [ka]

[0009] However, X is

[0010] [ka]

[0011] and R is a group that contains a radioactive metal ion or an optically imageable group; L 1 is a chemical bond,

[0012] [ka]

[0013] and L 2 teeth,

[0014] [ka]

[0015] and L 3 teeth,

[0016] [ka]

[0017] It is.

[0018] In one embodiment, in the peptide urea derivative represented by formula I, its pharma- ceutically acceptable salt, its solvate, or its pharma- ceutically acceptable salt, the de?nitions of certain groups are as shoWn beloW, and the remaining groups are as de?ned in any other embodiment (hereinafter abbreviated as “in one embodiment”).

[0019] X is

[0020] [ka]

[0021] and R is a group containing a radioactive metal ion or an optically imageable group; L 1 is a chemical bond,

[0022] [ka]

[0023] And L 2 teeth,

[0024] [ka]

[0025] And L 3 teeth,

[0026] [ka]

[0027] It is.

[0028] In one embodiment, the L 1 The carbonyl end of

[0029] [ka]

[0030] The L 2 Carbonyl end is L 1 The L 3 Carbonyl end is L 2 is linked to.

[0031] In one embodiment, L 1 is a chemical bond,

[0032] [ka]

[0033] and L 2 teeth,

[0034] [ka]

[0035] and L 3 teeth,

[0036] [ka]

[0037] and L 1 The C-terminus of

[0038] [ka]

[0039] The L 2 The e-terminus of L 1 The L 3 The g-terminus of 2 is linked to.

[0040] In one embodiment, L 1 is a chemical bond,

[0041] [ka]

[0042] And L 2 teeth,

[0043] [ka]

[0044] And L 3 teeth,

[0045] [ka]

[0046] and L 1 The C-terminus of

[0047] [ka]

[0048] The L2 The e-terminus of L 1 The L 3 The g-terminus of 2 is linked to.

[0049] In one embodiment, X is

[0050] [ka]

[0051] [ka]

[0052] The a-terminus of X is linked to R, and the two b-terminus of X are each linked to two L 3 is linked to.

[0053] In one embodiment, X is

[0054] [ka]

[0055] The a-terminus of X is linked to R, and the two b-terminus of X are each linked to two L 3 is linked to.

[0056] In one embodiment, the group containing a radioactive metal ion is composed of a radioactive metal ion and a group having a function of chelating a metal ion, and the radioactive metal ion is chelated to the group having a function of chelating a metal ion.

[0057] In one embodiment, the group having a function of chelating a metal ion is

[0058] [ka]

[0059] [ka]

[0060] It is.

[0061] In one embodiment, the group having a function of chelating a metal ion is

[0062] [ka]

[0063] It is.

[0064] In one embodiment, the group having a function of chelating a metal ion is

[0065] [ka]

[0066] It is.

[0067] In one embodiment, the radioactive metal ion has one or more of the following effects: (1)PET imaging; (2) SPECT imaging; (3) Radiation therapy. In one embodiment, the radioactive metal ion has one or more of the following effects: (1) Tracer; (2) Delivery; (3) Imaging; (4) Treatment.

[0068] In one embodiment, the radioactive metal ion is an α, β or γ emitting radioactive metal ion.

[0069] In one embodiment, the radioactive metal ion is68 Ga, 89 Zr, 64 Cu, 86 Y, 99m Tc, 111 In, 90 Y, 67 Ga, 177 Lu, 211 At, 153 Sm, 186 Re, 188 Re, 67 Cu, 212 Pb, 225 Ac, 213 Bi, 212 Bi or 212 It is Pb.

[0070] In one embodiment, the radioactive metal ion is 68 Ga 3+ , 89 Zr 4+ , 64 Cu 2+ , 86 Y 3+ , 99m Tc 4+ , 111 In 3+ , 90 Y, 67 Ga 3+ , 177 Lu 3+ , 211 At 3+ , 153 Sm 3+ , 186 Re 3+ , 188 Re 3+ , 67 Cu 2+ , 212 Pb 2+ , 225 Ac 3+ , 213 Bi 3+ , 212 Bi or 212 Pb 2+ It is.

[0071] In one embodiment, the radioactive metal ion is 68 Ga or 177 This is Lu.

[0072] In one embodiment, the radioactive metal ion is 68 Ga 3+ or 177 Lu 3+ It is.

[0073] In one embodiment, the group containing a radioactive metal ion is any one of the following groups:

[0074] [ka]

[0075] In one embodiment, the peptide urea derivative of Formula I has the structure:

[0076] [ka]

[0077] Here, M is a radioactive metal ion, and the radioactive metal ion is a trivalent radioactive metal ion, for example, 68 Ga 3+ , 86 Y 3+ , 111 In 3+ , 67 Ga 3+ , 177 Lu 3+ , 211 At 3+ , 153 Sm 3+ , 186 Re 3+ , 188 Re 3+ , 225 Ac 3+ or 213 Bi 3+ and preferably 68 Ga 3+ or 177 Lu 3+ It is.

[0078] In one embodiment, the optically imageable moiety is a fluorescent moiety, for example cy3, cy5 or cy7.

[0079] In one embodiment, the peptide urea derivative of formula I is a mixture of compound A and the radioactive metal ion (e.g., 68 Ga 3+ or 177 Lu 3+ ) and the structure of compound A is as shown in any one of the following formulas.

[0080] [ka]

[0081] [ka]

[0082] [ka]

[0083] [ka]

[0084] [ka]

[0085] [ka]

[0086] In one embodiment, the peptide urea derivative of formula I is a compound A. 177 Lu 3+ The structure of compound A is as shown in any one of the following formulae:

[0087] [ka]

[0088] [ka]

[0089] [ka]

[0090] [ka]

[0091] [ka]

[0092] [ka]

[0093] [ka]

[0094] [ka]

[0095] In one embodiment, the peptide urea derivative of formula I is a compound A. 68 Ga 3+ The structure of compound A is as shown in any one of the following formulae:

[0096] [ka]

[0097] [ka]

[0098] [ka]

[0099] [ka]

[0100] [ka]

[0101] [ka]

[0102] [ka]

[0103] The present invention further provides a method for preparing a peptide urea derivative of formula I above, comprising the step of chelating a radioactive metal ion with a compound of formula II.

[0104] [ka]

[0105] In the compound represented by the formula II, R' is a group having a function of chelating a metal ion.

[0106] The present invention further provides compounds of formula II.

[0107] [ka]

[0108] where R' is a group having a function of chelating a metal ion, and X and L 1 , L 2 and L 3 is as defined above.

[0109] In the compound represented by formula II, the function of chelating the metal ion is as defined above.

[0110] In the compound represented by formula II, the group having a function of chelating a metal ion is not chelated with a metal ion.

[0111] The structure of the compound represented by formula II is as shown in any one of the following formulae:

[0112] [ka]

[0113] [ka]

[0114] [ka]

[0115] [ka]

[0116] [ka]

[0117] [ka]

[0118] The present invention further provides the compounds shown below.

[0119] [ka]

[0120] [ka]

[0121] [ka]

[0122] [ka]

[0123] [ka]

[0124] The present invention further provides a pharmaceutical composition comprising a substance X which is a peptide urea derivative of formula I above, a pharma- ceutically acceptable salt thereof, a solvate thereof or a solvate of a pharma- ceutically acceptable salt thereof, and pharmaceutical auxiliaries.

[0125] In one embodiment, the pharmaceutical composition may be a pharmaceutical composition for treating or diagnosing prostate cancer.

[0126] In one embodiment, the pharmaceutical composition may be a pharmaceutical composition for imaging prostate cancer.

[0127] In one embodiment, the prostate cancer is castration-resistant prostate cancer.

[0128] In one embodiment, the prostate cancer is metastatic castration-resistant prostate cancer.

[0129] In one embodiment, the prostate cancer is PSMA positive prostate cancer. In one embodiment, the substance X is a therapeutically effective amount of substance X.

[0130] The present invention further provides the use of a substance X, which is a peptide urea derivative of formula I above, a pharma- ceutically acceptable salt thereof, a solvate thereof or a solvate of a pharma- ceutically acceptable salt thereof, in the manufacture of a medicament.

[0131] The medicament is a medicament for treating or diagnosing prostate cancer, or the medicament is a medicament for imaging prostate cancer.

[0132] In one embodiment, the medicament is a medicament for treating prostate cancer, and the radioactive metal ion is a gamma-ray emitting radioactive metal ion.

[0133] In one embodiment, the medicament is for treating prostate cancer, and the radioactive metal ion is 177 Lu 3+ It is.

[0134] In one embodiment, the medicament is a medicament for treating prostate cancer, and the radioactive metal ion is a radioactive metal ion that emits α or β rays.

[0135] In one embodiment, the medicament is a medicament for diagnosing prostate cancer, and the radioactive metal ion is 68 Ga 3+ or 64 Cu 2+ It is.

[0136] In one embodiment, the prostate cancer is castration-resistant prostate cancer.

[0137] In one embodiment, the prostate cancer is metastatic castration-resistant prostate cancer.

[0138] In one embodiment, the prostate cancer is PSMA-positive prostate cancer.

[0139] The term "pharmaceutical acceptable salt" refers to a salt obtained by reacting a compound with a pharmaceutically acceptable (relatively non-toxic, safe, and suitable for use by a patient) acid or base. When a 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. Pharmaceutically acceptable base addition salts include, but are not limited to, sodium salts, potassium salts, calcium salts, aluminum salts, magnesium salts, bismuth salts, ammonium salts, and the like. When a 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. Pharmaceutically acceptable acid addition salts include, but are not limited to, hydrochloride salts, sulfate salts, methanesulfonate salts, acetate salts, trifluoromethanesulfonate salts, and the like. For more information, see Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P. Heinrich Stahl, 2002).

[0140] The term "solvate" refers to a substance formed after crystallization of a compound with a solvent, including but not limited to water, ethanol, etc. Solvates are classified as stoichiometric and non-stoichiometric solvates.

[0141] The term "solvate of a pharma- ceutically acceptable salt" refers to a substance formed by combining a compound with a pharma- ceutically acceptable (relatively non-toxic, safe, and suitable for patient use) acid or base, a solvent (including, but not limited to, water, ethanol, etc.), where the pharma- ceutically acceptable salt has the same meaning as the term "pharmaceutically acceptable salt" above, and the solvent may be stoichiometric or non-stoichiometric. Solvates of pharma- ceutically acceptable salts include, but are not limited to, hydrochloride monohydrates, and the like.

[0142] The term "alkyl" refers to straight or branched chain alkyl having a specified number of carbon atoms (e.g., C1-C6). Alkyl includes, but is not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-amyl, n-hexyl, n-heptyl, n-octyl, and the like.

[0143] The term "cycloalkyl" or "carbocycle" refers to a saturated cyclic group consisting only of carbon atoms, which may be monocyclic, bridged, or spirocyclic, having a specified number of carbon atoms (e.g., C3-C6). Cycloalkyl includes, but is not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

[0144] The term "aryl" or "aryl ring" refers to an aryl having a specified number of carbon atoms (e.g., C6-C 10 ) and refers to a cyclic group consisting of only carbon atoms, which may be monocyclic or fused rings, and at least one ring is aromatic (according to the Shock Soul rule). Aryl is attached to other fragments within the molecule through an aromatic or non-aromatic ring. Aryl is an aromatic group such as phenyl, naphthyl,

[0145] [ka]

[0146] These include, but are not limited to:

[0147] " in structural fragments

[0148] [ka]

[0149] " indicates that the structural fragment is attached to another fragment within the molecule via that site.

[0150] The term "pharmaceutical adjuvants" refers to excipients and additives used in the manufacture and formulation adjustment of pharmaceuticals, and refers to all substances contained in drug formulations except active ingredients. For details, you can refer to the Pharmacopoeia of the People's Republic of China (2020 edition) or the Handbook of Pharmaceutical Excipients (Raymond C Rowe, 2009).

[0151] The term "therapeutically effective amount" refers to an amount of a compound that is sufficient to effectively treat a disease when administered to a patient. The therapeutically effective amount varies depending on the compound, the type of disease, the severity of the disease, the age of the patient, etc., but can be adjusted by those skilled in the art according to the situation.

[0152] The term "patient" refers to any animal that has received or is about to receive treatment, preferably a mammal, and most preferably a human, including, but not limited to, cows, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, etc.

[0153] The term "treatment" refers to any of the following situations: (1) ameliorating one or more biological manifestations of a disease; (2) disrupting one or more points in the biological cascade leading to a disease; or (3) slowing the progression of the development of one or more biological manifestations of a disease.

[0154] The term "prevention" refers to reducing the risk of developing a disease.

[0155] The present invention uses the following abbreviations: DMF stands for N,N-dimethylformamide. DMAP stands for 4-dimethylaminopyridine. Fmoc represents a 9-fluorenylmethoxycarbonyl protecting group. H-Glu(OtBu)-OH represents L-glutamic acid-5-tert-butyl ester. Lys stands for L-lysine. ivDde represents 1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl.

[0156] Each of the above preferred conditions can be arbitrarily combined to obtain each preferred embodiment of the present invention, without violating the ordinary skill in the art.

[0157] The reagents and materials used in the present invention are commercially available.

[0158] The positive effects of the present invention are: the derivative can be used not only for preoperative imaging diagnosis and grading of PSMA-positive prostate cancer, but also for the treatment of various types and stages of prostate cancer, realizing the integration of diagnosis and treatment, and is expected to be widely used. [Brief description of the drawings]

[0159] [Figure 1] 1 shows Micro-SPECT / CT imaging images of DB-01 in Mouse 1 at different time points. [Diagram 2] 1 shows Micro-SPECT / CT imaging images of DB-01 in Mouse 2 at different time points. [Diagram 3] In Efficacy Example 4, the uptake of different DB compounds in the tumor samples of mice 24 hours after administration is shown. [Figure 4] In Effect Example 4, the uptake in liver samples of mice 24 hours after administration of different DB compounds is shown. [Diagram 5] In Efficacy Example 4, the uptake in kidney samples of mice 24 hours after administration of different DB compounds is shown. [Figure 6] In Efficacy Example 4, the uptake of different DB compounds in the blood samples of mice 24 hours after administration is shown. [Figure 7] In Efficacy Example 4, the uptake in intramuscular samples of mice is shown 24 hours after administration of different DB compounds. [Figure 8] 1 shows competitive binding of DB and reference compounds on PSMA expressing cells. [Figure 9] 1 shows killing experiments of DB and reference compounds in LNCaP cells. [Figure 10] The lipid-water partition coefficients of the DB and reference compounds are shown. [Figure 11] 1 shows the albumin binding rate of DB and reference compounds in rat plasma. [Figure 12] 2 shows the efficacy (change in tumor volume) of DB and reference compounds in a 22RV1 tumor-bearing mouse animal model. [Figure 13] Figure 1 shows the efficacy (body weight change) of DB and reference compounds in a 22RV1 tumor-bearing mouse animal model. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0160] The present invention will be further described below in the form of examples, but the present invention is not limited to the scope of the examples. In the following examples, the experimental methods for which no specific conditions are described are selected according to the usual methods and conditions or product instructions.

[0161] Example 1 Synthesis of Compound M1

[0162] [ka]

[0163] General synthesis method A: Fmoc-Lys(ivDde)-Wang resin (0.3mmol / g) was added to a reaction vessel as the starting material, and 25% hexahydropyridine / DMF (volume ratio) was added and stirred for 30 minutes. The reaction solution showed a deep blue color when detected with ninhydrin. The reaction solution was suction filtered and washed five times with DMF to remove the N-terminal Fmoc protecting group and make the N-terminus a free amino. DMF was used as the solvent, and N,N-disuccinimidyl carbonate (1 equivalent), N,N-diisopropylethylamine DIPEA (2 equivalents) and 4-dimethylaminopyridine DMAP (2 equivalents) were added in a ratio of (1:2:2). The reaction was carried out under the protection of nitrogen gas for 1 hour, and H-Glu(OtBu)-OH (1.1 equivalents) was added and stirred for 24 hours. After removing the side chain Dde protecting group of Lys with 2% hydrazine hydrate / DMF solution, Fmoc-2-Nal-OH / HOBt / DIC (3 equivalents) was added to graft with resin to introduce 2-Nal amino acid residue. Next, Fmoc protecting group was removed with 25% hexahydropyridine / DMF (volume ratio) to make N-terminus of 2-Nal free amino group, then Fmoc-L-COOH / HOBt / N,N-diisopropyldiimine (DIC) (3 equivalents) was added to graft with resin to introduce L amino acid residue. Next, Fmoc protecting group was removed with 25% hexahydropyridine / DMF (volume ratio) to make N-terminus of L free amino group, and then synthesized to obtain product M1.

[0164] This method is a general synthesis method for compound M1, in which L is any group at the corresponding position of DB-01 to DB-10.

[0165] Example 2 Synthesis of Compound M2

[0166] [ka]

[0167] General synthesis method B: Using M1 as the starting material, the cleavage reagent (trifluoroacetic acid:H2O:triisopropylsilane=90:5:5, v / v) was used to dissolve the target polypeptide from compound M2 resin to remove the side chain protecting groups (cleavage at 30°C for 3 h). The filtrate was added to a large amount of cold anhydrous ethyl ether to precipitate and separate the polypeptide, which was then centrifuged. After washing with ethyl ether several times and drying, the polypeptide product M2 was obtained.

[0168] Example 3 Synthesis of Compound M3

[0169] [ka]

[0170] General synthesis method C: Using M1 as raw material, general synthesis method A was used to couple amino acid FmocNHXCOOH, and the Fmoc protecting group was removed to make the N-terminus of X a free amino group, and then the target polypeptide was dissolved from compound M3 resin using a cleavage reagent (trifluoroacetic acid:H2O:triisopropylsilane=90:5:5, v / v) to remove the side chain protecting group (cleavage at 30°C for 3 hours). The filtrate was added to a large amount of cold anhydrous ethyl ether to precipitate and separate the polypeptide, which was then centrifuged. After washing with ethyl ether several times and drying, the crude polypeptide product M3 was obtained.

[0171] This method is a general synthesis method for compound M3, in which X is any group at the corresponding position of DB-01 to DB-10.

[0172] Example 4 Synthesis of Compound M5

[0173] [ka]

[0174] General synthetic method D: 3,5-dicarboxyaniline (1.0 equiv.) was dissolved in DMF (50 mL), DIPEA (1.5 equiv.) was added, followed by NHS-activated DOTA ester (1.1 equiv.), the reaction was stirred at room temperature overnight, the reaction solution was poured into water, extracted with ethyl acetate (3×30 mL), the combined extracts were washed with saturated NaCl solution (30 mL), dried (Na2SO4), filtered, concentrated to remove the solvent, and then purified by column chromatography (SiO2, 5-10% MeOH / DCM) to give M4 (0.8 equiv., 80%). M4 (0.5 equiv), DMAP (0.1 equiv), N-hydroxysuccinimide (1.1 equiv) were dissolved in DMF (20 mL), reacted overnight, diluted with ethyl acetate (100 mL), washed with saturated NaHCO3 solution (35 mL), the organic phase was washed with saturated NaCl solution, dried (Na2SO4), filtered and concentrated to give M5. MS (ESI): 916. The crude M5 was used directly in the next step.

[0175] Example 5 Synthesis of Compound M6

[0176] [ka]

[0177] General synthetic method E: M5 (0.01 equiv.), M3 (0.02 equiv.), DIPEA (0.025 equiv.) were dissolved in DMF (10 mL) and reacted overnight, the solvent was removed under vacuum, the residue was stirred at 30 °C for 3 h using cleavage reagent (trifluoroacetic acid: HO: triisopropylsilane = 90: 5: 5, v / v), poured into a large amount of cold anhydrous ethyl ether, the precipitate was filtered, washed with cold anhydrous ethyl ether, dried and purified by preparative liquid chromatography (C18 column, A: 100% water / 0.1% TFA, B: 100% acetonitrile / 0.1% TFA), and the fractions containing the target product were mixed and lyophilized to obtain product M6.

[0178] Example 6 Synthesis of Compound M11

[0179] [ka]

[0180] Synthesis of compound M11 N-Boc glycine (1.0 equiv.) and DCC (1.3 equiv.) were added to THF (100 mL) and stirred at room temperature for 30 min., 1,3,5-triazinane (1.1 equiv.) was added, stirred overnight, concentrated, and the residue was purified by column chromatography (SiO2, 5-10% MeOH / DCM) to give M7 (0.8 equiv.). MS (ESI): 245 (M+1). + .

[0181] M7 (1.0 equiv.), tert-butyl 2-bromoacetate (2.1 equiv.), and triethylamine (2.5 equiv.) were dissolved in DMF (100 mL), the reaction solution was stirred at room temperature overnight, concentrated, and the residue was purified by column chromatography (SiO2, 5-10% MeOH / DCM) to give M8 (0.75 equiv.). MS (ESI): 473 (M+1). + .

[0182] M8 (1.0 equiv.) was mixed with a mixture of TFA and DCM (TFA:DCM:TIS=9:1:0.1), stirred at room temperature for 6 h, concentrated, washed with anhydrous ethyl ether, filtered, and the solid (M9) was dried in vacuum and used directly in the next step.

[0183] M9 (0.5 equiv.), DIPEA (0.5 equiv.), and NHS-activated DOTA ester (0.55 equiv.) were dissolved in DMF (50 mL) and general synthetic method D was used to obtain M10 (0.5 equiv.). MS (ESI): 816 (M+ 1). + .

[0184] M10 (0.5 equiv.), DMAP (0.1 equiv.), and N-hydroxysuccinimide (1.05 equiv.) were dissolved in DMF (20 mL) and general synthetic method D was used to obtain M11 (0.48 equiv.). MS (ESI): 1010 (M + 1) + .

[0185] Example 7 Synthesis of Compound M12

[0186] [ka]

[0187] 3,5-Bis-BOC-aminobenzoic acid (0.5 eq.), DCC (0.55 eq.) and DMAP (0.1 eq.) were dissolved in DMF (100 mL) and stirred at room temperature for 30 min., then DOTA derivative (0.5 eq.) was added, the reaction solution was stirred at room temperature overnight, concentrated, filtered, the filtrate was concentrated, the residue was mixed with a mixture of TAF in DCM (TFA:DCM:TIS=9:1:0.1), stirred at room temperature for 2 h, concentrated, washed with anhydrous ethyl ether, filtered, and the solid (M12) was dried in vacuum to obtain M12. MS (ESI): 581 (M+1) + M12 was used directly in the next step without purification, i.e., the preparation of DB-07.

[0188] Example 8 Synthesis of compound M13

[0189] [ka]

[0190] DOTA derivative (0.15 equiv), DCC (0.15 equiv), DMAP (0.05 equiv) were dissolved in DMF (20 mL) and stirred for 30 min, then tris(2-aminoethyl)amine (0.5 equiv) was added, the reaction solution was stirred overnight, concentrated, diluted with DCM, filtered and concentrated to give crude M13, which was used directly in the next reaction. MS(ESI): 701 (M+1). + .

[0191] Example 9 Synthesis of compound M14 (general synthesis method F):

[0192] [ka]

[0193] 1,3,5-Triacryloylhexahydro-1,3,5-triazine (0.5 equiv.) and M3c (0.15 equiv.) were stirred in DMF overnight, concentrated, and the residue was diluted with cold ethyl ether, precipitated, filtered, washed with cold ethyl ether, and dried to give M14. MS (ESI): 882 (M+1). + .

[0194] Example 10 Synthesis of Compounds Synthesis of M2a:

[0195] [ka]

[0196] Compound M2a was obtained using general synthetic methods A and B. MS (ESI): 656 (M + 1) + .

[0197] Synthesis of M3a:

[0198] [ka]

[0199] Compound M3a was obtained using general synthetic methods A and C. MS (ESI): 713 (M + 1) + .

[0200] Synthesis of M3b:

[0201] [ka]

[0202] Compound M3b was obtained using general synthetic method A and the corresponding C. MS (ESI): 804 (M + 1) + .

[0203] Synthesis of M3c:

[0204] [ka]

[0205] Compound M3c was obtained using general synthetic methods A and C. MS (ESI): 730 (M + 1) + .

[0206] M3d compositing:

[0207] [ka]

[0208] Compound M3d was obtained using general synthetic methods A and C. MS (ESI): 776 (M+1) + .

[0209] Synthesis of M3e:

[0210] [ka]

[0211] Compound M3e was obtained using general synthetic methods A and C. MS (ESI): 579 (M + 1) + .

[0212] Synthesis of M3f:

[0213] [ka]

[0214] Compound M3f was obtained using general synthetic methods A and B. MS(ESI): 676(M+1) + .

[0215] Example 11 Synthesis of DB-01:

[0216] [ka]

[0217] DB-01 was prepared from M5 and M2a according to general synthetic method D. MS (ESI): 1844 (M + 1) + .

[0218] Example 12 Synthesis of DB-02:

[0219] [ka]

[0220] DB-02 was prepared from M5 and M3a according to general synthetic method D. MS (ESI): 1844 (M + 1) + .

[0221] Example 13 Synthesis of DB-03:

[0222] [ka]

[0223] DB-03 was prepared from M11 and M3a according to general synthetic method E. MS(ESI): 1923 (M + 1) + .

[0224] Example 14 Synthesis of DB-04:

[0225] [ka]

[0226] Compound DB-04 was synthesized according to the following synthesis route. First, M15 was prepared from lysine derivative and M2a according to synthesis method E. After removing the Boc protection on the nitrogen atom of M15 with TFA, M16 was prepared together with DOTA derivative according to synthesis method E. After removing the Fmoc protecting group on M16, it was coupled with M3d. Then, the Boc protecting group on DOTA was removed with TFA to obtain DB-04 crude product. The product was purified by preparative liquid chromatography and lyophilized to obtain DB-04 purified product. MS(ESI):1909 (M + 1) + .

[0227] [ka]

[0228] Example 15 Synthesis of DB-05:

[0229] [ka]

[0230] DB-05 was prepared by coupling M13 with M3b. MS(ESI): 2105 (M + 1) + .

[0231] Example 16 Synthesis of DB-06:

[0232] [ka]

[0233] DB-06 was prepared from M14 and M3c according to general synthetic method F. MS (ESI): 2173 (M + 1) + .

[0234] Example 17 Synthesis of DB-07:

[0235] [ka]

[0236] DB-07 was prepared by coupling M12 with M3d. MS (ESI): 2097 (M + 1) + .

[0237] Example 18 Synthesis of DB-08:

[0238] [ka]

[0239] DB-08 was prepared by the synthetic method of DB-07. MS (ESI): 2098 (M + 1) + .

[0240] Example 19 Synthesis of DB-09:

[0241] [ka]

[0242] DB-09 was produced by the same method as DB-08. MS (ESI): 1703 (M + 1) + .

[0243] Example 20 Synthesis of DB-010:

[0244] [ka]

[0245] DB-10 was prepared by the synthesis method of DB-04. MS (ESI): 1949 (M + 1) + .

[0246] Example 21 Metal Complexes 177 Synthesis of Lu-DB-01:

[0247] [ka]

[0248] 1) 4.1 g of sodium acetate was weighed out, 45 mL of ultrapure water was added, and after complete dissolution, the pH was adjusted to 4.0 with glacial acetic acid, and then 50 mL of ultrapure water was added to obtain a 1 M sodium acetate solution. The metal bath reactor was opened and preheated to 95°C.

[0249] 2) Precursor compound DB-01 (1 mg) was weighed out, and 1000 μL of sodium acetate solution was added to each of them, and completely dissolved to obtain a precursor solution of 1 μg / μL. 10 μL (10 μg) of the precursor solution was taken and further diluted with 240 μL of sodium acetate solution. Next, 250 μCi of each was added. 177 The LuCl3 solution was added, mixed uniformly, and placed in a reactor at 95°C for reaction for 30 minutes, and purified by C18 Sep-Pak.

[0250] 3) 0.5 g of EDTA sodium salt was weighed out, and 50 mL of physiological saline was added and completely dissolved to obtain a 1% aqueous solution of EDTA sodium salt. 177 The LuCl3 solution was taken and spotted 1 cm from the bottom of a silica gel plate for flash thin layer chromatography and dried. 2 μL of the reacted solution was taken and spotted 1 cm from the bottom of a silica gel plate for flash thin layer chromatography and dried. 0.5 mL of 1% EDTA sodium salt solution was used as the developer, and the bottom of the silica gel plate was placed on the developer in a glass test tube, and the bottom of the silica gel plate was extended to the liquid surface of the developer not more than 5 mm. A rubber stopper was placed on top, and when the developer had developed to 9-10 cm, the silica gel plate was removed and dried. The silica gel plate was scanned with a gamma scanner, 177 The solutions after reaction with LuCl3 solution were compared, and the radiochemical purity and labeling rate were calculated from the peak areas.

[0251] As shown in Table 1, 177 The LuCl3 solution was spread in the developer to the top of the silica gel plate.

[0252] [Table 1]

[0253] As shown in Table 2, the labeled compounds 177 Lu-DB-01 could only be developed to the bottom of the silica gel plate in the developer, and its radiochemical purity was 99.1% and the labeling rate was 100%.

[0254] [Table 2]

[0255] Example 22: Synthesis of DB-11

[0256] [ka]

[0257] DB-11 is prepared according to the following general synthetic method: a series of condensation reactions involving solid-phase synthesis affords intermediate JH-04S10-01 (MS: [M + H] + = 489.61) and DB-11-01 (MS: [M + H] + = 788.50), and DB-11-02 (MS: [M / 2 + H] + = 1015.43), which was deprotected to give DB-11-03 (MS: [M / 2 + H] + = 852.81), which was reacted with DOTA-NHS to give the final product compound DB-11. HPLC: 98.54%, RT: 8.23; MS: [M / 2+H] + =1046.36.

[0258] [ka]

[0259] [ka]

[0260] Example 23: Synthesis of DB-12

[0261] [ka]

[0262] DB-12 was synthesized according to the following general method. DB-A1 (MS: [M + H] + = 816.83) and DB-A2 (MS: [M + H] + = 681.73) was coupled to DB-A3 (MS: [M / 2 + H] + = 1139.49) and after two deprotections, intermediate DB-A5 (MS: [M / 2 + H] + = 916.21) was reacted with DOTA-NHS to obtain DB-12. HPLC: 95.58%, RT: 10.963; MS: [M / 2+H] + =1109.90.

[0263] [ka]

[0264] Example 24: Synthesis of DB-13

[0265] [ka]

[0266] DB-13 was prepared by referring to the general synthesis method of DB-12. HPLC: 97.73%, RT: 9.214; MS: [M / 2 +H] + =1047.34.

[0267] Example 25: Synthesis of DB-14

[0268] [ka]

[0269] DB-14 was prepared by referring to the general synthesis method of DB-12. HPLC: 95.8%, RT: 9.52; MS: [M / 2 + H] + = 1047.34.

[0270] Example 26: Synthesis of DB-15

[0271] [ka]

[0272] DB-15 was synthesized with reference to general methods D and E. HPLC: 95.8%, RT: 12.34; MS: [M / 2+H] + =1124.59.

[0273] Example 27: Synthesis of DB-16

[0274] [ka]

[0275] DB-16 was synthesized by referring to the general method of DB-11. HPLC: 96.62%, RT: 9.543; MS: [M / 2+H] + = 1012.36.

[0276] Example 28: Synthesis of DB-17

[0277] [ka]

[0278] DB-17 was synthesized with reference to general methods D and E. HPLC: 99.24%, RT: 10.423; MS: [M / 2+H] + = 937.27.

[0279] Example 29: Synthesis of DB-18

[0280] [ka]

[0281] DB-18 was synthesized with reference to general methods D and E. HPLC: 95.64%, RT: 7.78; MS: [(M-1) / 3-1] + =626.37.

[0282] Example 30: Synthesis of DB-19

[0283] [ka]

[0284] DB-19 was synthesized with reference to general methods D and E. HPLC: 97.73%, RT: 9.10; MS: [M / 2+H] + =1028.17.

[0285] Example 31: Synthesis of DB-20

[0286] [ka]

[0287] DB-20 was synthesized by referring to the general method of DB-11. HPLC: 98.95%, RT: 11.294; MS: [M / 2+H] + =1074.49.

[0288] Example 32: Synthesis of DB-21

[0289] [ka]

[0290] DB-21 was synthesized with reference to general methods D and E. HPLC: 98.01%, RT: 12.446; MS: [M / 2+H] + = 1100.24.

[0291] The structures of the reference compounds used in the following Effect Examples are as follows:

[0292] [ka]

[0293] Effect Example 1 Measurement of binding affinity and dissociation constant of candidate molecules targeting PSMA Dissociation constants were determined by Biacore 8K (GE Healthcare) according to the instructions provided by the instrument manufacturer. Briefly, PSMA-bio protein diluted in PBS pH 7.4, 1 mM TCEP, 0.05% Tween 20, 2% DMSO buffer was immobilized on the flow cell of the SA sensor chip. Five series of diluted concentrations of candidate molecules targeting PSMA were injected into the flow cell at 30 μL / min using PBS pH 7.4, 1 mM TCEP, 0.05% Tween 20, 2% DMSO as the running buffer, in which the binding time was 90 s. The buffer flow for dissociation was maintained for 1800 s. The KD values ​​of the interaction between the candidate small molecules and PSMA protein were evaluated using the fitting models of 1.0 and 1:1 binding in the Biacore 8K evaluation software.

[0294] The results are shown in Table 3. The binding affinity of DB-01 was slightly higher than that of PSMA-617.

[0295] [Table 3]

[0296] Effect Example 2 Micro-SPECT / CT scanning imaging experiment of mice transplanted with human prostate cancer cell line 22RV1 Human prostate cancer cell line 22RV1 cells were subcutaneously injected into the armpits of nude mice to establish a prostate cancer tumor model. 177 Lu-labeled compound [ 177 Lu]DB-01 was injected intravenously at approximately 3.7MBq (100μCi / 200μL), and SPECT / CT images were collected using U-SPECT+ / CT (MI Labs) at 3, 5, 24, 72, 120 and 168 hours after injection, and quantitative analysis of the obtained SPECT / CT image results was performed.

[0297] Table 4 shows the 177 The results of quantitative Micro-SPECT / CT analysis, expressed as the uptake in each organ of mice, i.e., quantitative % injected dose (ID) / g, are shown at 3, 5, 24, 72, 120, and 168 h after injection of [Lu]DB-01.

[0298] As shown in Table 4, 177 Lu]DB-01 was excreted very rapidly from the kidney and bladder early after injection.

[0299] [ 177 For [Lu]DB-01, tumor uptake increased over time, with a calculated tumor uptake of 13.55±3.85%ID / g at 180 min, and tumor residence time was significantly increased, with tumor uptake still at 7.85±1.16%ID / g at 168 h. 177 For all of the [Lu]DB-01 compounds, the uptake by the kidney decreased from 5 hours onwards and they were rapidly excreted from the body.

[0300] [Table 4]

[0301] As can be seen from Table 4 and Figs. 1 and 2, 177After Lu-DB-01 entered the mouse body, it was rapidly distributed to various organs and target sites, and its main route of metabolism was urination. After 5 hours, the concentration of the radioactive drug in the tumor reached its highest point and then gradually decreased. After 24 hours, in addition to the accumulation of the radioactive drug in the tumor, the accumulation of the radioactive drug in other organs was also significantly reduced. After 7 days, a significant amount of the radioactive drug still existed in the tumor. This result showed that the compound disclosed in this patent application not only has better PSMA cell affinity compared with the reference compound PSMA-617, but also has better pharmacokinetics, making it a molecule with greater potential as an integrated nuclear drug for diagnosis and treatment.

[0302] Effect Example 3 Affinity test of DB compound for PSMA protein using Biacore Ligand binding of PSMA protein (Sino biological) was detected using a Biacore 8K (Cytiva) instrument. PSMA protein was captured on a SA chip. Prior to ligand immobilization (channels 1, 2, flow rate: 10 μL / min), PSMA protein (10 μg / ml, flow rate: 5 μL / min, injection time: 600 s) was immobilized in channel 2 using flow buffer and three consecutive injections of 1 M NaCl in 50 mM NaOH were performed to condition the sensor surface. After each ligand injection, an additional wash with 1 M NaCl and isopropanol in 50 mM NaOH (channels 1, 2, flow rate: 10 μL / min, injection time: 60 s) was included.

[0303] All compounds were dissolved in 100% dimethyl sulfoxide, diluted to 10 mM, and then further diluted to the appropriate maximum concentration with assay buffer (PBS, pH 7.4, 1 mM TCEP (tris-(2-hydroxyethyl)phosphine), 0.05% P20, 2% dimethyl sulfoxide). Analyses were performed at an assay temperature of 15 °C, with assay steps set to all LMW kinetics; cycle type = single cycle (contact time: 90 s, separation time: 1800 s, flow rate: 30 μL / min, flow paths 1,2); flow path detection = 2-1, etc. Data evaluation was performed using Biacore Insight evaluation software, and the data were fitted to a 1:1 binding model.

[0304] The Biacore results are as shown in Table 5: pKD = -LogKD, where KD was the binding affinity of the compound for the PSMA protein measured by Biacore. KD (M) was expressed as Kd (1 / s) / Ka (1 / M s). A represented pKD > 8, B represented 8 < pKD < 7, C represented 7 < pKD < 6, and D represented pKD > 6.

[0305]

Table 5

[0306] Example of Effect 4 177 Lu Labeling and In Vivo Distribution Experiment in Animals Tissue distribution of PSMA-positive tumor-bearing mice: Tumor-bearing mice (purchased from Shanghai Alamo Pharmaceutical Technology Co., Ltd.), 3×10 6 individual 22rv1 cells (in 50% matrix gel, Corning) were subcutaneously inoculated into the right shoulder of approximately 6-9-week-old (Balb / c Nude) mice. When the tumors had grown to an appropriate size, 177Lu radiolabeled DB compound was injected via the tail vein of mice (approximately 1.85 MBq to 3.7 MBq / mouse, specific activity: 22423.82 KBq / μg), and the animals were euthanized using carbon dioxide inhalation 24 h after administration. After euthanasia, blood and organs (blood, liver, kidney, muscle, and tumor) were collected.

[0307] Blood was collected from the abdominal aorta and immediately weighed into a designated centrifuge tube at 100 μL. After organs were harvested, they were washed twice with deionized water, wiped, placed in pre-weighed test tubes, weighed again, and the weight of the sample was calculated and the weight of the sample was measured on the day of collection. A gamma counter was used to measure the radioactivity counts of all blood and tissue samples, and the results are shown in Figure 3, Figure 4, Figure 5, Figure 6, Figure 7 and Table 6.

[0308] After intravenous administration, 177 The Lu-labeled DB compounds were rapidly distributed in each organ of tumor-bearing mice, and the uptake in non-target organs was always low. They were rapidly metabolized to the outside of the body, and the blood clearance rate was high. They were mainly metabolized in the kidney and excreted from the body. The uptake of the Lu-177-labeled compounds into the tumors remained at a high level even 24 hours after administration, and DB-12 and DB-19 in particular had an abnormally high uptake in PSMA-positive 22RV1 tumor cells.

[0309] [Table 6]

[0310] Effect Example 5 Competitive Binding (1) Prepare a cell suspension of 22Rv1 cells in logarithmic growth phase at a cell density of 1 × 10 4 1 mL of the solution was inoculated into a 24-well cell culture plate and cultured overnight in a 37°C incubator. (2) The cell culture medium was aspirated, the cells were washed once with PBS, and 975 μL of additive-free medium was added. (3) To each well, 25 μL of a fixed concentration of radiolabeled PSMA-617 ligand (final concentration in medium: 2 μCi / mL) and different concentrations (final concentrations in medium: 1000 ng / ml, 100 ng / ml, 10 ng / ml, 1 ng / ml, 0.1 ng / ml, 0.01 ng / ml, 0.001 ng / ml, 0 ng / ml) of the unlabeled blocking compounds to be tested, DB-18, DB-19, DB-13, and E-3, were added. (4) The cells were incubated on or off ice for 2 hours. (5) The cells were washed three times with ice-cold PBS. (6) The cells were lysed with 0.5 mL of 1 M sodium hydroxide and washed twice with 0.5 mL of PBS. Sodium hydroxide (0.5 mL) and PBS (0.5 mL x 2) solutions were collected and the uptake was measured.

[0311] The experimental results are shown in FIG. 8, and the results show that the IC50 values ​​of compounds E-3, DB-13, DB-18 and DB-19 are 2.06 nM, 1.83 nM, 2.58 nM and 0.93 nM, respectively.

[0312] Effect Example 6 Cell Death A. Logarithmic growth phase LNCaP cells were prepared in a cell suspension at a cell density of 1 x 10 5 1 mL of the solution was inoculated into a 24-well cell culture plate and cultured in a 37°C incubator for 24 hours. B. Using serum-containing medium, 177Lu-labeled compounds (177Lu-E-3) were prepared at radioactive concentrations of 5, 15, and 45 μCi / mL for use (for unlabeled E-3, the precursor concentration corresponds to 45 μCi / mL). C. The above 177Lu-labeled compound solution was added at 1 mL / well to the experimental group, and serum-containing medium was added at 1 mL / well to the control group. D. The cells were cultured in an incubator for 24 hours, and then the medium was replaced with fresh medium. E. On day 5, enhanced CCK8 kit was used to detect the proliferation of cells in each group.

[0313] The experimental results are shown in Figure 9. The results show that at the tested doses, both the two compounds and the reference compound have cell-killing effects and have a dose-response relationship. At an equivalent dose of 45 μCi / mL, DB-13 showed a stronger cell-killing effect, with the cell-killing rate reaching 67%.

[0314] Effect Example 7 Measurement of cLogP Three EP tubes were taken, and 0.1 ml of saturated n-octanol and 80 μl of ultrapure water were added to each, followed by addition of 20 μl (approximately 1 Mbq) of Lu-177 labeled DB-13, DB-18 and DB-19 compound solutions, followed by shaking uniformly and then centrifuging at room temperature (2000 r / min, 5 min, centrifugal radius 10 cm). 100 μl each was taken from the lipid layer and aqueous layer of each tube, the number of radioactive phases per minute was measured, LogP was calculated, and the average value was used as the result.

[0315] The experimental results are shown in FIG. 10. The experimental logP values ​​of DB-13, DB-18, DB-19 and E-3 were −1.96, −1.86, −1.79 and −3.46, respectively, and the results indicated that all of the test compounds had good water solubility.

[0316] Effect Example 8 Test of PPB Labeled 177 Lu-E-3, 177 Lu-DB-13, 177 Lu-DB-18 and 177 Lu-DB-19 compound (1 μCi) was taken and 50 μL of PBS was added to obtain a 20 μCi / mL reaction solution. 50 μL of the 20 μCi / mL reaction solution was added to 200 μL of plasma, repeated for three tubes, and incubated at room temperature for 10 minutes after uniform mixing, and the sample was added to the ultrafiltration tube and centrifuged at 13000 rpm for 45 minutes, then 50 μL of saline was added and centrifugation was continued for 15 minutes, and the upper chamber and filtrate were counted separately.

[0317] PPB=[(top chamber count−background count)] / (filtrate count+top chamber count−2×background count)]×100%.

[0318] The experimental results are shown in FIG. 11. Comparing the synthesized compounds with the reference compounds, all had higher albumin binding rates except for DB-13 (DB-13: 90.84%, E-3: 86.61%), while the albumin binding rates of DB-18 and DB-19 were all lower than the reference compounds, at 70.51% and 59.07%, respectively.

[0319] Effect Example 9 Treatment Approximately 6-8 week-old (Balb / c nude) mice were injected with 3 × 10 6 Cells 22rv1 (50% Matrigel, Corning) were inoculated subcutaneously. When the tumors had grown to the size required for the experiment, the animals were randomly divided into 11 experimental groups according to tumor volume, with 5 animals in each group. The animals' weights and tumor sizes were measured, and the animals were divided into control group (saline), group 1 ( 177 Lu-DB-13 0.15mCi / mouse), group 2 ( 177 Lu-DB-13 0.45mCi / mouse), group 3 ( 177 Lu-DB-13 0.05mCi / mouse), group 4 ( 177 Lu-B-11 0.05mCi / mouse), group 5 ( 177 Lu-DB-11 0.15mCi / mouse), group 6 ( 177 Lu-DB-11 0.45mCi / mouse), group 7 ( 177 Lu-PSMA-617 0.15mCi / mouse), group 8 ( 177 Lu-PSMA-617 0.45mCi / mouse), group 9 ( 177 Lu-DB-19 0.05mCi / mouse), group 10 ( 177 Lu-DB-19 0.15mCi / mouse), group 11 ( 177 Dosing was initiated according to the Lu-DB-19 0.45mCi / animal standard deviation (D = 0.001), and observations of general health and appearance were performed daily after the start of the experiment, and animals were weighed and tumor sizes were measured prior to each sampling time point. Any abnormal observations found throughout the study should be recorded in the raw data.

[0320] The experimental results are shown in Figure 12, Figure 13 and Table 7. The results show that compared with PSMA-617, DB-19 showed excellent tumor growth inhibitory effects on tumor cells at two different doses, medium and high (0.15 mCi and 0.45 mCi), and DB-11 had a certain tumor growth inhibitory effect after administration at a high dose (0.45 mCi), which decreased over time, but had a higher inhibitory effect than PSMA-617.

[0321] [Table 7]

Claims

1. A peptide urea derivative represented by formula I, a pharma- ceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharma- ceutically acceptable salt thereof. 【Chemistry 1】 (However, X is 【Chemistry 2】 and R is a group that contains a radioactive metal ion or an optically imageable group; L １ is a chemical bond, 【Chemistry 3】 and L ２ teeth, 【Chemistry 4】 and L ３ teeth, 【Chemistry 5】 It is.)

2. 2. A peptide urea derivative represented by formula I according to claim 1, a pharma- ceutically acceptable salt thereof, a solvate thereof or a solvate of a pharma- ceutically acceptable salt thereof. (X is 【Chemistry 6】 and R is a group that contains a radioactive metal ion or an optically imageable group; L １ is a chemical bond, 【Chemistry 7】 And L ２ teeth, 【Chemistry 8】 And L ３ teeth, 【Chemistry 9】 It is.)

3. Said L １ The carbonyl end of 【Chemistry 10】 is connected to said L ２ The carbonyl end of L １ is connected to said L ３ The carbonyl end of L ２ 3. A peptide urea derivative of formula I according to claim 1 or 2, a pharma- ceutically acceptable salt thereof, a solvate thereof or a solvate of a pharma- ceutically acceptable salt thereof, characterized in that it is linked to

4. (1) L １ is a chemical bond, 【Chemistry 11】 And L ２ teeth, 【Chemistry 12】 And L ３ teeth, 【Chemistry 13】 and the L １ The C-terminus of 【Chemistry 14】 is connected to said L ２ The e-terminus of L １ is connected to said L ３ The g-terminus of ２ is connected to Preferably, L １ is a chemical bond, 【Chemistry 15】 And L ２ teeth, 【Chemistry 16】 And L ３ teeth, 【Chemistry 17】 and the L １ The C-terminus of 【Chemistry 18】 is connected to said L ２ The e-terminus of L １ is connected to said L ３ The g-terminus of ２ is connected to (2) X is, 【Chemistry 19】 【Chemistry 20】 The a-terminus of X is linked to R, and the two b-terminus of X are each linked to two L ３ is connected to Preferably, X is 【Chemistry 21】 The a-terminus of X is linked to R, and the two b-terminus of X are each linked to two L ３ is connected to A peptide urea derivative of formula I according to claim 1, a pharma- ceutically acceptable salt thereof, a solvate thereof or a solvate of a pharma- ceutically acceptable salt thereof, characterized in that it satisfies one or more of the following conditions:

5. The group containing a radioactive metal ion is composed of a radioactive metal ion and a group having a function of chelating a metal ion, and the radioactive metal ion is chelated with the group having a function of chelating the metal ion. For example, the group having a function of chelating a metal ion is 【Chemical 22】 【Chemistry 23】 and, for example, the group having a function of chelating a metal ion is 【Chemistry 24】 and preferably 【Chemistry 25】 3. The peptide urea derivative of claim 1 or 2, which is represented by formula I, a pharma- ceutically acceptable salt thereof, a solvate thereof or a solvate of a pharma- ceutically acceptable salt thereof.

6. (1) The radioactive metal ion has one or more of the following effects:

1. PET imaging; 2. SPECT imaging; 3. radiotherapy; (2) The radioactive metal ion has one or more of the following effects:

1. tracer, 2. delivery, 3. imaging, 4. therapy; (3) The radioactive metal ion is ６８ G.A. ８９ Zr, ６４ Cu, ８６ Y. ９９ｍ Tc, １１１ In, ９０ Y. ６７ G.A. １７７ Lu, ２１１ At, １５３ Sm, １８６ Re, １８８ Re, ６７ Cu, ２１２ Pb, ２２５ A c, ２１３ Bi, ２１２ Bi or ２１２ Pb, preferably ６８ Ga or １７７ Lu, for example, the radioactive metal ion is ６８ G ３＋ , ８９ Zr ４＋ , ６４ Cu ２＋ , ８６ Y ３＋ , ９９ｍ Tc ４＋ , １１１ In ３＋ , ９０ Y. ６７ G ３＋ , １７７ Lu ３＋ , ２１１ At, １５３ Sm, １８６ Re, １８８ Re, ６７ Cu ２＋ , ２１２ Pb ２＋ , ２２５ A.C. ３＋ , ２１３ Bi ３＋ , ２１２ Bi or ２１２ Pb ２＋ and preferably ６８ G ３＋ or １７７ Lu ３＋ That is, 6. A peptide urea derivative of formula I according to claim 5, a pharma- ceutically acceptable salt thereof, a solvate thereof or a solvate of a pharma- ceutically acceptable salt thereof, characterized in that it satisfies one or more of the following conditions:

7. The peptide urea derivative represented by formula I according to claim 1 or 2, characterized in that the group containing a radioactive metal ion is any one of the following groups, a pharma- ceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharma- ceutically acceptable salt thereof: 【Chemistry 26】

8. The peptide urea derivative represented by formula I according to claim 1 or 2, characterized in that it has the structure of the following formula: 【Chemical 27】 (wherein M is a radioactive metal ion, the radioactive metal ion is a trivalent radioactive metal ion, for example, ６８ G ３＋ , ８６ Y ３＋ , １１１ In ３＋ , ６７ G ３＋ , １７７ Lu ３＋ , ２１１ At ３＋ , １５３ Sm ３＋ , １８６ R ３＋ , １８８ R ３＋ , ２２５ A.C. ３＋ or ２１３ Bi ３＋ and preferably ６８ G ３＋ or １７７ Lu ３＋ It is.)

9. The peptide urea derivative represented by formula I is a compound A and １７７ Lu ３＋ and a compound formed by chelating said compound A with said compound A, and the structure of said compound A is as shown in any one of the following formulas: 【Chemistry 28】 【Chemical Formula 29】 【Chemistry 30】 【Chemistry 31】 【Chemistry 32】 【Chemical 33】

10. The peptide urea derivative represented by formula I is a compound A and ６８ G ３＋ and a compound formed by chelating said compound A with said compound A, and the structure of said compound A is as shown in any one of the following formulas: 【Chemical 34】 【Chemistry 35】 【Chemical 36】 【Chemical 37】 【Chemical Formula 38】 【Chemical 39】

11. A process for the preparation of a peptide urea derivative of formula I according to any one of claims 1 to 10, comprising a step of chelating a radioactive metal ion with a compound of formula II. 【Chemistry 40】 (In the compound represented by formula II, R' is a group having a function of chelating a metal ion.)

12. A compound represented by formula II. 【Chemistry 41】 (wherein R' is a group having a function of chelating a metal ion, and X, L １ , L ２ and L ３ is as defined in any one of claims 1 to 10.

13. The compound of formula II according to claim 12, wherein the definition of the group having a function of chelating a metal ion in the compound of formula II is as defined in claim 5.

14. The compound represented by formula II according to claim 13, characterized in that the compound represented by formula II is: 【Chemistry 42】 【Chemistry 43】 【Chemistry 44】 【Chemistry 45】 【Chemistry 46】 【Chemistry 47】 【Chemistry 48】

15. A compound characterized in that it is as shown in the following formula: 【Chemistry 49】 【Chemistry 50】 【Chemistry 51】

16. A pharmaceutical composition comprising a substance X which is a peptide urea derivative of formula I according to any one of claims 1 to 10, a pharma- ceutically acceptable salt thereof, a solvate thereof or a solvate of a pharma- ceutically acceptable salt thereof, and pharmaceutical auxiliaries.

17. Use of a substance X which is a peptide urea derivative of formula I according to any one of claims 1 to 10, a pharma- ceutically acceptable salt thereof, a solvate thereof or a solvate of a pharma- ceutically acceptable salt thereof in the manufacture of a medicament, comprising The medicament is a medicament for treating or diagnosing prostate cancer, or the medicament is a medicament for imaging prostate cancer.

18. 18. The use of claim 17, wherein the prostate cancer is castration-resistant prostate cancer, metastatic castration-resistant prostate cancer or PSMA-positive prostate cancer.

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