Peptidourea derivative, pharmaceutical composition containing peptidourea derivative and application of peptidourea derivative

Abstract

The invention discloses a peptidourea derivative, a pharmaceutical composition containing the peptidourea derivative and application of the peptidourea derivative. The derivative is shown as a formula I. The derivative can be used for preoperative development diagnosis and grading of PSMA positive prostate cancer, can also be used for treatment of various types and stages of prostate cancer, achieves diagnosis and treatment integration, and has a wide application prospect. # imgabs0 #

Description

[0001] This application is a divisional application of Chinese patent application No. 2023100673528, filed on January 30, 2023. The invention of the original application is entitled: A peptide urea derivative, a pharmaceutical composition containing the same, and its application. The original application claims priority to Chinese patent application No. 2022101162309, filed on January 30, 2022. Technical Field

[0002] The invention relates to a peptide urea derivative, a pharmaceutical composition containing the same and application thereof. Background Art

[0003] Prostate cancer is a highly prevalent cancer both worldwide and in China.

[0004] Early imaging diagnosis and treatment of prostate cancer has become an urgent problem in China and even in the world. Prostate cancer begins to occur in the tissues around the prostate, and as it grows, it gradually metastasizes to other important organs such as the lungs and bones. In the early stages, there are no obvious symptoms, but as prostate cancer grows, it causes problems such as urethral compression and urinary tract obstruction, and further metastasizes to the spine or pelvis. For the diagnosis of prostate cancer, imaging diagnostic methods such as SPECT (single photon emission computed tomography) and PET (positron emission tomography) are currently being used. The principle is to use radioactive isotopes that emit gamma rays or positrons to label PSMA-targeted polypeptide substances, and to display the presence and distribution of tumor cells in tomographic images and three-dimensional images through prostate cancer-specific targeted distribution. These imaging diagnostic methods have recently been greatly promoted due to the development of SPECT-CT / MRI and PET-CT / MRI combined with CT or MRI, which have greatly improved image quality. The radiopharmaceuticals currently used for specific imaging of prostate cancer use PSMA ligands as targeting groups, which 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 a specific molecular marker for prostate cancer. It is expressed in very small amounts in kidney, small intestine, and brain tissues, and the expression level in tumor tissues is much higher than that in normal tissues. Representative ligands for PSMA are peptide derivatives such as Glu-urea-Lys (GUL) or Glu-urea-Cys (GUC). Therefore, by labeling a radioactive isotope with such a peptide ligand, the prepared radiopharmaceutical can be used for PET or SPECT imaging of prostate cancer, or for the treatment of prostate cancer (Meder, et al., Bioconjugate Chem 2012, 23: 688-697). The radioactive isotopes used to label peptides are mainly radionuclides that emit α rays, radionuclides that emit β rays, radionuclides that emit γ rays, and radionuclides that emit positron beams. Among them, alpha-ray emitting radionuclides and beta-ray emitting radionuclides are used for treatment, while gamma-ray emitting radionuclides and positron beam emitting radionuclides are used for diagnosis through nuclear imaging. There are generally two methods for labeling ligands with radioisotopes: a method in which the ligand is directly bound to the radioisotope, or a method in which the ligand is chelated to the radioisotope by 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, 131I, etc.The method using a bifunctional chelating agent (BFCA) is mainly used for labeling various metal radioisotopes. The type of the bifunctional chelating agent (BFCA) can be selected according to the properties of the ligand and the radioisotope.

[0005] Currently, castration surgery, anti-androgen castration methods, and androgen receptor inhibitors are the mainstream treatment options for prostate cancer. Although these treatment options are very effective in the initial stage, a large number of patients will develop castration-resistant prostate cancer (CRPC) or even metastatic castration-resistant prostate cancer (mCRPC). mCRPC is a disease with limited treatment options and significant unmet medical needs, so radioactive drugs targeting PSMA have become a research hotspot in recent years.

[0006] A series of clinical studies have been conducted on the use of radiopharmaceuticals targeting PSMA to treat mCRPC patients. Although the preliminary clinical results of radiopharmaceuticals such as 177Lu-PSMA-617 and 177Lu-PSMA I&T are encouraging, there are also some problems. For example, nearly 30% of patients do not respond to this treatment method. One possible explanation is that not enough radiopharmaceuticals are delivered to tumor lesions due to unsatisfactory pharmacokinetic. Another concern is the long-term accumulation of radiopharmaceuticals such as 177Lu-PSMA-617 in organs such as kidneys and salivary glands. Therefore, a radiopharmaceutical targeting PSMA with high activity, high selectivity, and better pharmacokinetics is a continuous hot spot in the treatment and diagnosis of mCRPC. Summary of the invention

[0007] The technical problem to be solved by the present invention is that the existing peptide urea derivatives have a single structure, and the uptake of radiopharmaceuticals by non-targeted organs and the long-term retention of the drugs in them, such as in the kidneys, are not high or the retention time is not long enough in the desired targeted tumor. To this end, the present invention provides a peptide urea derivative, a pharmaceutical composition containing it, 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 the treatment of prostate cancer of various types and stages, achieving integrated diagnosis and treatment, and has broad application prospects. 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 radiopharmaceuticals in targeted tumors are greatly improved.

[0008] The present invention provides a peptide urea derivative as shown in formula I, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof;

[0009]

[0010] Among them, X is

[0011] R is a group containing a radioactive metal ion or a group capable of optical imaging;

[0012] L 1 For chemical bonds,

[0013] L 2 for

[0014] L 3 for

[0015] In a certain embodiment, in the peptide urea derivative as shown in Formula I, its pharmaceutically acceptable salt, its solvate or its pharmaceutically acceptable salt solvate, the definitions of certain groups are as follows, and the definitions of the remaining groups are as described in any other embodiment (hereinafter referred to as "in a certain embodiment"):

[0016] X is

[0017] R is a group containing a radioactive metal ion or a group capable of optical imaging;

[0018] L 1 For chemical bonds, L 2 for L 3 for

[0019] In one embodiment, the L 1 The carbonyl end connected, the L 2 The carbonyl terminal and L 1 connected, the L 3 The carbonyl terminal and L 2 connected.

[0020] In one scheme, L 1 For chemical bonds,

[0021] L 2 for

[0022]

[0023] L 3 for

[0024] The L 1 The C-terminus of connected, the L 2 The e end and L 1 connected, the L 3 The g end and L 2 connected.

[0025] In one scheme, L 1 For chemical bonds, L 2 for L 3 for

[0026] The L 1 The C-terminus of connected, the L 2 The e end and L 1 connected, the L 3 The g end and L 2 connected.

[0027] In one solution, X is

[0028] The a end of X is connected to R, and the two b ends of X are connected to two L 3 connected.

[0029] In one solution, X is

[0030] The a end of X is connected to R, and the two b ends of X are connected to two L 3 connected.

[0031] In a certain embodiment, the group containing a radioactive metal ion is composed of a radioactive metal ion and a group having a function of chelating the metal ion, and the radioactive metal ion is chelated with the group having a function of chelating the metal ion.

[0032] In one embodiment, the group having the function of chelating metal ions is

[0033]

[0034] In one embodiment, the group having the function of chelating metal ions is

[0035] In one embodiment, the group having the function of chelating metal ions is

[0036] In one embodiment, the radioactive metal ion has one or more of the following effects:

[0037] (1) PET imaging;

[0038] (2) SPECT imaging;

[0039] (3)Radiotherapy.

[0040] In one embodiment, the radioactive metal ion has one or more of the following effects:

[0041] (1) Tracing;

[0042] (2) delivery;

[0043] (3) Imaging;

[0044] (4) Treatment.

[0045] In one embodiment, the radioactive metal ion is a radioactive metal ion that releases α, β or γ rays.

[0046] In one embodiment, the radioactive metal ion is 68 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 Bior 212 Pb.

[0047] 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 Bior 212 Pb 2 + .

[0048] In one embodiment, the radioactive metal ion is 68 Ga or 177 Lu.

[0049] In one embodiment, the radioactive metal ion is 68 Ga 3+ or 177 Lu 3+ .

[0050] In one embodiment, the group containing a radioactive metal ion is any of the following groups:

[0051]

[0052] In one embodiment, the peptide urea derivative as shown in formula I has the following structure:

[0053]

[0054] Wherein, M is a radioactive metal ion; 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+ , preferably 68 Ga 3+ or 177 Lu 3+ .

[0055] In one embodiment, the group capable of optical imaging may be a fluorescent group, such as cy3, cy5 or cy7.

[0056] In one embodiment, the peptide urea derivative as shown in formula I is a compound A and a radioactive metal ion (eg 68 Ga 3+ or 177 Lu 3+ ) is a compound formed by chelation, wherein the structure of the compound A is shown in any of the following:

[0057]

[0058]

[0059]

[0060]

[0061]

[0062]

[0063] In one embodiment, the peptide urea derivative as shown in formula I is compound A and 177 Lu 3+ The compound formed by chelation, the structure of the compound A is shown as any one of the following:

[0064]

[0065]

[0066]

[0067]

[0068]

[0069]

[0070] In one embodiment, the peptide urea derivative as shown in formula I is compound A and 68 Ga 3+ The compound formed by chelation, the structure of the compound A is shown as any one of the following:

[0071]

[0072]

[0073]

[0074]

[0075]

[0076]

[0077]

[0078] The present invention also provides a method for preparing the peptide urea derivative as shown in formula I, which comprises the following steps: chelating a radioactive metal ion with a compound as shown in formula II;

[0079]

[0080] In the compound shown in Formula II, R' is a group having the function of chelating metal ions.

[0081] The present invention also provides a compound as shown in formula II

[0082]

[0083] Among them, R' is a group with the function of chelating metal ions, X, L 1 , L 2 and L 3 The definition of is as mentioned above.

[0084] In the compound shown in Formula II, the group having the function of chelating metal ions can be defined as described above.

[0085] In the compound shown in Formula II, the group having the function of chelating metal ions does not chelate with metal ions.

[0086] The structure of the compound shown in Formula II can be any of the following:

[0087]

[0088]

[0089]

[0090]

[0091]

[0092]

[0093]

[0094] The present invention also provides a compound as shown below:

[0095]

[0096]

[0097]

[0098] The present invention also provides a pharmaceutical composition, which comprises a substance X and a pharmaceutical excipient; the substance X is the above-mentioned peptide urea derivative as shown in Formula I, a pharmaceutically acceptable salt thereof, a solvate thereof or a solvate of a pharmaceutically acceptable salt thereof.

[0099] In a certain embodiment, the pharmaceutical composition may be a pharmaceutical composition for treating or diagnosing prostate cancer.

[0100] In one embodiment, the pharmaceutical composition may be a pharmaceutical composition for imaging prostate cancer.

[0101] In one embodiment, the prostate cancer is castration-resistant prostate cancer.

[0102] In one embodiment, the prostate cancer is metastatic castration-resistant prostate cancer.

[0103] 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.

[0104] The present invention also provides a use of a substance X in the preparation of a drug; the substance X is the above-mentioned peptide urea derivative as shown in formula I, a pharmaceutically acceptable salt thereof, a solvate thereof, or a solvate of a pharmaceutically acceptable salt thereof;

[0105] The drug is a drug for treating or diagnosing prostate cancer, or the drug is a drug for imaging prostate cancer.

[0106] In one embodiment, the drug is a drug for treating prostate cancer, and the radioactive metal ions are radioactive metal ions that release gamma rays.

[0107] In one embodiment, the drug is a drug for treating prostate cancer, and the radioactive metal ion is 177 Lu 3+ .

[0108] In one embodiment, the drug is a drug for diagnosing prostate cancer, and the radioactive metal ion is a radioactive metal ion that emits α or β rays.

[0109] In one embodiment, the drug is a drug for diagnosing prostate cancer, and the radioactive metal ion is 68 Ga 3+ or 64 Cu 2+ .

[0110] In one embodiment, the prostate cancer is castration-resistant prostate cancer.

[0111] In one embodiment, the prostate cancer is metastatic castration-resistant prostate cancer.

[0112] In one embodiment, the prostate cancer is PSMA-positive prostate cancer.

[0113] The term "pharmaceutically acceptable salt" refers to a salt obtained by reacting a compound with a pharmaceutically acceptable (relatively non-toxic, safe, and suitable for use by patients) acid or base. When the compound contains a relatively acidic functional group, a base addition salt can be obtained by contacting the free form of the compound with a sufficient amount of a pharmaceutically acceptable base in a suitable inert solvent. 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 the compound contains a relatively basic functional group, an acid addition salt can be obtained by contacting the free form of the compound with a sufficient amount of a pharmaceutically acceptable acid in a suitable inert solvent. Pharmaceutically acceptable acid addition salts include, but are not limited to, hydrochlorides, sulfates, methanesulfonates, acetates, trifluoromethylsulfonates, and the like. For details, see Handbook of Pharmaceutical Salts: Properties, Selection, and Use (P. Heinrich Stahl, 2002).

[0114] 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 divided into stoichiometric solvates and non-stoichiometric solvates.

[0115] The term "solvate of a pharmaceutically acceptable salt" refers to a substance formed by combining a compound with a pharmaceutically acceptable (relatively non-toxic, safe, and suitable for use by patients) acid or base, and a solvent (including but not limited to water, ethanol, etc.), wherein the pharmaceutically acceptable salt has the same meaning as the term "pharmaceutically acceptable salt" above, and the solvent is stoichiometric or non-stoichiometric. The solvate of a pharmaceutically acceptable salt includes but is not limited to hydrochloride monohydrate.

[0116] The term "alkyl" refers to a straight or branched chain alkyl group having a specified number of carbon atoms (e.g., C1 to C6). Alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like.

[0117] The term "cycloalkyl" or "carbocycle" refers to a saturated cyclic group consisting only of carbon atoms with a specified number of carbon atoms (e.g., C3 to C6), which is a monocyclic, bridged or spirocyclic ring. Cycloalkyl includes, but is not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.

[0118] The term "aryl" or "aromatic ring" refers to a group having a specified number of carbon atoms (e.g., C6 to C 10) is a cyclic group consisting only of carbon atoms, which is a single ring or a condensed ring, and at least one ring is aromatic (in accordance with Huckel's rule). The aryl group is connected to other fragments in the molecule through an aromatic ring or a non-aromatic ring. Aryl groups include but are not limited to phenyl, naphthyl, wait.

[0119] In the structure fragment It means that the structural fragment is connected to other fragments in the molecule through this site.

[0120] The term "pharmaceutical excipients" refers to excipients and additives used in the production of drugs and the preparation of prescriptions. It is all substances contained in drug preparations except active ingredients. For details, please refer to the Pharmacopoeia of the People's Republic of China (2020 edition) or Handbook of Pharmaceutical Excipients (Raymond C Rowe, 2009).

[0121] The term "therapeutically effective amount" refers to the amount of a compound administered to a patient that is sufficient to effectively treat a disease. The therapeutically effective amount will vary 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 as appropriate.

[0122] The term "patient" refers to any animal that has been or is about to be treated, preferably a mammal, most preferably a human. Mammals include, but are not limited to, cows, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, etc.

[0123] The term "treat" refers to any of the following: (1) alleviating one or more biological manifestations of a disease; (2) interfering with one or more points in the biological cascade that initiates a disease; or (3) slowing the progression of one or more biological manifestations of a disease.

[0124] The term "prevention" refers to reducing the risk of developing a disease.

[0125] The present invention uses the following abbreviations:

[0126] DMF stands for N,N-dimethylformamide.

[0127] DMAP stands for 4-dimethylaminopyridine.

[0128] Fmoc represents a 9-fluorenylmethoxycarbonyl protecting group.

[0129] H-Glu(OtBu)-OH represents L-glutamic acid-5-tert-butyl ester.

[0130] Lys means L-lysine.

[0131] ivDde represents 1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl.

[0132] Without violating the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

[0133] The reagents and raw materials used in the present invention are commercially available.

[0134] The positive and progressive effect of the present invention is that the derivative can be used for preoperative imaging diagnosis and grading of PSMA-positive prostate cancer, and can also be used for the treatment of prostate cancer of various types and stages, achieving integrated diagnosis and treatment, and has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0135] Figure 1 Micro-SPECT / CT imaging images of DB-01 in mouse 1 at different times.

[0136] Figure 2 Micro-SPECT / CT images of DB-01 in mouse 2 at different times.

[0137] Figure 3 In Effect Example 4, the uptake of samples in the tumors of mice was measured 24 hours after the mice were administered with different DB compounds.

[0138] Figure 4 In Effect Example 4, the uptake of samples in the liver of mice was measured 24 hours after the mice were administered with different DB compounds.

[0139] Figure 5 In Effect Example 4, the uptake of samples in the kidneys of mice was measured 24 hours after the mice were administered with different DB compounds.

[0140] Figure 6 In Effect Example 4, the uptake of samples in the blood of mice was measured 24 hours after the mice were administered with different DB compounds.

[0141] Figure 7 In Example 4, the intramuscular uptake of samples was measured 24 hours after the mice were administered with different DB compounds.

[0142] Figure 8 Competitive binding of DB and reference compounds on PSMA-expressing cells.

[0143] Fig. 9 This is a killing experiment of DB and reference compounds on LNCaP cells.

[0144] Fig.10 is the lipid-water partition coefficient of DB and reference compound.

[0145] Fig.11 is the albumin binding rate of DB and reference compound in rat plasma.

[0146] Fig.12 The pharmacodynamics (tumor volume changes) of DB and reference compounds in the 22RV1 tumor-bearing mouse model.

[0147] Fig.13 The drug effects (body weight changes) of DB and reference compounds on 22RV1 tumor-bearing mouse model. DETAILED DESCRIPTION

[0148] The present invention is further described below by way of examples, but the present invention is not limited to the scope of the examples. The experimental methods in the following examples without specifying specific conditions are carried out according to conventional methods and conditions, or selected according to the product specifications.

[0149] Example 1 Synthesis of Compound M1

[0150]

[0151] General synthesis method A: Fmoc-Lys(ivDde)-Wang resin (0.3 mmol / g) is used as the starting material, added to the reaction vessel, and then 25% hexahydropyridine / DMF (volume ratio) is added, and stirred for 30 minutes. The ninhydrin detection shows a dark blue color, and the reaction liquid is drained and filtered, and washed with DMF 5 times to remove the N-terminal Fmoc protecting group to make the N-terminal a free amino group; DMF is used as the solvent, N, N-disuccinimidyl carbonate (1 equivalent), N, N-diisopropylethylamine DIPEA (2 equivalents) and 4-dimethylaminopyridine DMAP (2 equivalents) are added in a ratio of (1:2:2), and the reaction is carried out under nitrogen protection for 1 hour, and then H-Glu(OtBu)-OH (1.1 equivalents) is added and stirred for 24 hours. After removing the Dde protecting group of the side chain of Lys with 2% hydrazine hydrate / DMF solution, Fmoc-2-Nal-OH / HOBt / DIC (3 equivalents) was added to the resin for grafting and introducing the 2-Nal amino acid residue. Subsequently, the Fmoc protecting group was removed again with 25% hexahydropyridine / DMF (volume ratio) to make the N-terminus of 2-Nal a free amino group, and Fmoc-L-COOH / HOBt / N, N-diisopropyldiimide (DIC) (3 equivalents) was added to the resin for grafting and introducing the L amino acid residue. Subsequently, the Fmoc protecting group was removed again with 25% hexahydropyridine / DMF (volume ratio) to make the N-terminus of L a free amino group, and the product M1 was synthesized.

[0152] This method is a general synthesis method for compound M1, wherein L is a group at the corresponding position in any DB-01 to DB-10.

[0153] Example 2 Synthesis of Compound M2

[0154]

[0155] General synthesis method B: Using M1 as the starting material, use a cleavage reagent (trifluoroacetic acid: H2O: triisopropylsilane = 90:5:5, v / v) to cleave the target polypeptide from the compound M2 resin and remove the side chain protecting group (cleavage at 30°C for 3 hours). Add the filtrate to a large amount of cold anhydrous ether to precipitate the polypeptide, and centrifuge. Wash with ether several times and dry to obtain the polypeptide product M2.

[0156] Example 3 Synthesis of Compound M3

[0157]

[0158] General Synthesis Method C: Using M1 as the raw material, using General Synthesis Method A, coupling the amino acid FmocNHXCOOH, removing the Fmoc protecting group to make the N-terminus of X a free amino group, and then using a cleavage reagent (trifluoroacetic acid: H2O: triisopropylsilane = 90:5:5, v / v) to cleave the target polypeptide from the compound M3 resin and remove the side chain protecting group (cleavage at 30°C for 3 hours). The filtrate was added to a large amount of cold anhydrous ether to precipitate the polypeptide, and centrifuged. After washing with ether several times and drying, the crude polypeptide M3 was obtained.

[0159] This method is a general synthesis method for compound M3, wherein X is a group at a corresponding position in any of DB-01 to DB-10.

[0160] Example 4 Synthesis of Compound M5

[0161]

[0162] General Synthesis Method D: 3,5-Dicarboxyaniline (1.0 eq.) was dissolved in DMF (50 mL), DIPEA (1.5 eq.) was added, followed by NHS-activated DOTA ester (1.1 eq.), and the reaction was stirred at room temperature overnight. The reaction solution was poured into water and extracted with ethyl acetate (3 x 30 mL). The combined extracts were washed with saturated NaCl solution (30 mL), dried (Na2SO4), filtered, concentrated to remove solvent, and purified by column chromatography (SiO2, 5-10% MeOH / DCM) to give M4 (0.8 eq., 80%). M4 (0.5 equivalent), DMAP (0.1 equivalent), N-hydroxysuccinimide (1.1 equivalent) 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 reaction.

[0163] Example 5 Synthesis of Compound M6

[0164]

[0165] General synthetic method E: M5 (0.01 equivalent), M3 (0.02 equivalent), and DIPEA (0.025 equivalent) were dissolved in DMF (10 mL), and the reaction was allowed to proceed overnight. The solvent was removed under vacuum, and the residue was stirred with a cutting reagent (trifluoroacetic acid: H2O: triisopropylsilane = 90:5:5, v / v) at 30°C for 3 hours, poured into a large amount of cold anhydrous ether, and the precipitate was filtered, washed with cold anhydrous ether, dried, and purified by preparative liquid phase (C18 column, A: 100% water / 0.1% TFA, B: 100% acetonitrile / 0.1% TFA), and the fractions containing the target product were mixed and freeze-dried to obtain the product M6.

[0166] Example 6 Synthesis of Compound M11

[0167]

[0168] Synthesis of compound M11

[0169] N-Boc glycine (1.0 equivalent) and DCC (1.3 equivalent) were added to THF (100 mL), stirred at room temperature for 30 minutes, 1,3,5-triazinane (1.1 equivalent) was added, stirred overnight, concentrated, and the residue was purified by column chromatography (SiO2, 5-10% MeOH / DCM) to obtain M7 (0.8 equivalent), MS (ESI): 245 (M+1) + .

[0170] M7 (1.0 equivalent), tert-butyl 2-bromoacetate (2.1 equivalent), and triethylamine (2.5 equivalent) 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 obtain M8 (0.75 equivalent), MS (ESI): 473 (M+1) + .

[0171] M8 (1.0 equivalent) was mixed with a mixture of TAF and DCM (TFA:DCM:TIS=9:1:0.1), stirred at room temperature for 6 hours, concentrated, washed with anhydrous ether, filtered, and the solid (M9) was directly used in the next reaction after vacuum drying.

[0172] M9 (0.5 eq.), DIPEA (0.5 eq.), NHS-activated DOTA ester (0.55 eq.) were dissolved in DMF (50 mL) and M10 (0.5 eq.) was obtained using General Synthesis Method D. MS (ESI): 816 (M+1) + .

[0173] M10 (0.5 eq.), DMAP (0.1 eq.), N-hydroxysuccinimide (1.05 eq.) were dissolved in DMF (20 mL) and M11 (0.48 eq.) was obtained using the general method D. MS (ESI): 1010 (M+1)+.

[0175] Example 7 Synthesis of Compound M12

[0176]

[0177] 3,5-Bis-BOC-aminobenzoic acid (0.5 equivalent), DCC (0.55 equivalent) and DMAP (0.1 equivalent) were dissolved in DMF (100 mL) and stirred at room temperature for 30 minutes, then DOTA derivative (0.5 equivalent) was added, the reaction solution was stirred at room temperature overnight, concentrated, filtered, the filtrate was concentrated, the residue was mixed with TAF and DCM mixture (TFA: DCM: TIS = 9: 1: 0.1), stirred at room temperature for 2 hours, concentrated, washed with anhydrous ether, filtered, and the solid (M12) was dried in vacuo 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.

[0178] Example 8 Synthesis of Compound M13

[0179]

[0180] DOTA derivative (0.15 equivalent), DCC (0.15 equivalent) and DMAP (0.05 equivalent) were dissolved in DMF (20 mL) and stirred for 30 minutes, followed by the addition of tris(2-aminoethyl)amine (0.5 equivalent). The reaction solution was stirred overnight, concentrated, diluted with DCM, filtered, and concentrated to obtain crude M13, which was directly used in the next step. MS (ESI): 701 (M+1) + .

[0181] Example 9 Synthesis of Compound M14 (General Synthesis Method F):

[0182]

[0183] 1,3,5-Triacryloylhexahydro-1,3,5-triazine (0.5 equivalent) and M3c (0.15 equivalent) were stirred in DMF overnight, concentrated, the residual solution was diluted with cold ether, the precipitate was filtered, washed with cold ether, and dried to obtain M14. MS (ESI): 882 (M+1) + .

[0184] Example 10 Synthesis of Compounds

[0185] Synthesis of M2a:

[0186]

[0187] Compound M2a was obtained using general synthetic methods A and B, MS (ESI): 656 (M+1) + .

[0188] Synthesis of M3a:

[0189]

[0190] Compound M3a was obtained using general synthetic methods A and C, MS (ESI): 713 (M+1) + .

[0191] Synthesis of M3b:

[0192]

[0193] Compound M3b was obtained using general synthetic method A and corresponding C, MS (ESI): 804 (M+1) + . Synthesis of M3c:

[0194]

[0195] Compound M3c was obtained using general synthetic methods A and C, MS (ESI): 730 (M+1) + .

[0196] Synthesis of M3d:

[0197]

[0198] Compound M3d was obtained using general synthetic methods A and C, MS (ESI): 776 (M+1) + .

[0199] M3e Synthesis:

[0200]

[0201] Compound M3e was obtained using general synthetic methods A and C, MS (ESI): 579 (M+1) + .

[0202] Synthesis of M3f:

[0203]

[0204] Compound M3f was obtained using general synthetic methods A and B, MS (ESI): 676 (M+1) + .

[0205] Embodiment 11 Synthesis of DB-01:

[0206]

[0207] DB-01 was prepared from M5 and M2a according to general synthesis method D, MS (ESI): 1844 (M+1) + .

[0208] Example 12 Synthesis of DB-02:

[0209]

[0210] DB-02 was prepared from M5 and M3a according to general synthesis method D, MS (ESI): 1844 (M+1) + .

[0211] Synthesis of Example 13DB-03:

[0212]

[0213] DB-03 was prepared from M11 and M3a according to general synthesis method E. MS (ESI): 1923 (M+1) + .

[0214] Embodiment 14 Synthesis of DB-04:

[0215]

[0216] Compound DB-04 is prepared according to the following synthetic route: first, a lysine derivative and M2a are reacted according to the synthetic method E to obtain M15, and then the Boc protection on the nitrogen atom of M15 is removed by TFA and then coupled with a DOTA derivative according to the synthetic method E to obtain M16, and the Fmoc protecting group on M16 is removed and coupled with M3d, and then the Boc protecting group on DOTA is removed by TFA to obtain crude DB-04, which is purified by preparative liquid phase and freeze-dried to obtain pure DB-04. MS (ESI): 1909 (M+1) + .

[0217]

[0218] Embodiment 15 Synthesis of DB-05:

[0219]

[0220] DB-05 was prepared by coupling M13 and M3b. MS (ESI): 2105 (M+1) + .

[0221] Example 16 Synthesis of DB-06:

[0222]

[0223] DB-06 was prepared from M14 and M3c according to the general synthesis method F. MS (ESI): 2173 (M+1) + .

[0224] Example 17 Synthesis of DB-07:

[0225]

[0226] DB-07 was prepared by coupling M12 and M3d. MS (ESI): 2097 (M+1) + .

[0227] Embodiment 18 Synthesis of DB-08:

[0228]

[0229] DB-08 was prepared by the method of synthesizing DB-07. MS (ESI): 2098 (M+1) + .

[0230] Embodiment 19 Synthesis of DB-09:

[0231]

[0232] DB-09 was synthesized using the method for preparing DB-08: MS (ESI): 1703 (M+1)+ .

[0233] Embodiment 20 Synthesis of DB-010:

[0234]

[0235] DB-10 was prepared by the method of synthesizing DB-04. MS (ESI): 1949 (M+1) + .

[0236] Example 21 Metal Complex 177 Lu-DB-01 Synthesis:

[0237]

[0238] 1) Weigh 4.1 g of sodium acetate, add 45 mL of ultrapure water, and after it is completely dissolved, adjust the pH to 4.0 with glacial acetic acid, and then add ultrapure water to 50 mL to obtain a 1 M sodium acetate solution. Open the metal bath reactor and preheat it to 95°C.

[0239] 2) Weigh the precursor compound DB-01 (1 mg) and add 1000 μL of sodium acetate solution to each of them. After fully dissolving, a 1 μg / μL precursor solution is obtained. Take 10 μL (10 μg) of the precursor solution and add 240 μL of sodium acetate solution to dilute it. Then add 250 μCi of 177 The LuCl3 solution was mixed and placed in a reactor at 95°C for 30 minutes and purified using C18 Sep-Pak.

[0240] 3) Weigh 0.5 g of EDTA sodium salt, add 50 mL of physiological saline, and fully dissolve to obtain a 1% EDTA sodium salt aqueous solution. 177 Spot 2μL of LuCl3 solution on the instant thin layer chromatography silica gel plate 1cm from the bottom and blow dry. Take 2μL of the solution after the reaction, spot it on the instant thin layer chromatography silica gel plate 1cm from the bottom and blow dry. Use 0.5mL of 1% EDTA sodium salt solution as the developing agent, place the bottom of the silica gel plate in the developing agent in the glass test tube, and the bottom of the silica gel plate should not extend more than 5mm into the developing agent liquid surface. Cover with a rubber stopper, take out the silica gel plate when the developing agent is developed to 9-10cm, and blow dry. Scan the silica gel plate with a γ scanner and compare 177 The radiochemical purity and labeling rate of LuCl3 solution and the solution after reaction were calculated by peak area.

[0241] As shown in Table 1, 177 LuCl3 solution was developed in the developing agent on the top of the silica gel plate.

[0242] Table 1: Region: 177 Lu detector:PMT

[0243]

[0244] Total area:2401Counts

[0245] Average background: 0Counts

[0246] As shown in Table 2, the labeled compounds 177 Lu-DB-01 can only be developed to the bottom of the silica gel plate in the developing agent, and its radiochemical purity is 99.1% and the labeling rate is also 100%.

[0247] Table 2: Region: 177 Lu detector:PMT

[0248]

[0249] Total area:2498Counts

[0250] Average background: 0Counts

[0251] Example 22: Synthesis of DB-11

[0252]

[0253] DB-11 was prepared by the following general synthesis method: a series of condensation reactions, including solid phase synthesis, were used to obtain the intermediate JH-04S10-01 (MS: [M+H] + =489.61) and DB-11-01 (MS: [M+H] + =788.50), and DB-11-02 was obtained by condensation of JH-04S10-01 and DB-11-01 (MS: [M / 2+H] + =1015.43), and deprotected to give DB-11-03 (MS: [M / 2+H] + =852.81), the intermediate was reacted with DOTA-NHS to obtain the final product compound DB-11. HPLC: 98.54%, RT: 8.23; MS: [M / 2+H] + =1046.36.

[0254]

[0255] Example 23: Synthesis of DB-12

[0256]

[0257] DB-12 was synthesized by the following general method. DB-A1 (MS: [M+H] +=816.83) and DB-A2 (MS: [M+H] + =681.73) to give DB-A3 (MS: [M / 2+H] + =1139.49), after two deprotections, the intermediate DB-A5 (MS: [M / 2+H] + =916.21) was reacted with DOTA-NHS to give DB-12.

[0258] HPLC: 95.58%, RT: 10.963; MS: [M / 2+H] + =1109.90.

[0259]

[0260] Example 24: Synthesis of DB-13

[0261]

[0262] 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.

[0263] Example 25: Synthesis of DB-14

[0264]

[0265] 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.

[0266] Example 26: Synthesis of DB-15

[0267]

[0268] DB-15 was synthesized according to general methods D and E. HPLC: 95.8%, RT: 12.34; MS: [M / 2+H] + =1124.59.

[0269] Example 27: Synthesis of DB-16

[0270]

[0271] DB-16 was synthesized according to the general method of DB-11. HPLC: 96.62%, RT: 9.543; MS: [M / 2+H] + =1012.36.

[0272] Example 28: Synthesis of DB-17

[0273]

[0274] DB-17 was synthesized according to general methods D and E. HPLC: 99.24%, RT: 10.423; MS: [M / 2+H] + =937.27.

[0275] Example 29: Synthesis of DB-18

[0276]

[0277] DB-18 was synthesized according to general methods D and E. HPLC: 95.64%, RT: 7.78; MS: [(M-1) / 3-1] + =626.37.

[0278] Example 30: Synthesis of DB-19

[0279]

[0280] DB-19 was synthesized by referring to general methods D and E. HPLC: 97.73%, RT: 9.10; MS: [M / 2+H] + =1028.17.

[0281] Example 31: Synthesis of DB-20

[0282]

[0283] DB-20 was synthesized according to the general method of DB-11. HPLC: 98.95%, RT: 11.294; MS: [M / 2+H] + =1074.49.

[0284] Example 32: Synthesis of DB-21

[0285]

[0286] DB-21 was synthesized according to general methods D and E. HPLC: 98.01%, RT: 12.446; MS: [M / 2+H] + =1100.24.

[0287] The structures of the reference compounds used in the following examples are as follows:

[0288]

[0289] Effect Example 1 Determination of Binding Affinity and Dissociation Constant of Candidate Molecules Targeting PSMA

[0290] The dissociation constant was determined by Biacore 8K (GE Healthcare) following the instructions of the instrument provided by the manufacturer. In brief, PSMA-bio protein diluted in PBS pH 7.4, 1 mM TCEP, 0.05% Tween20, 2% DMSO buffer was immobilized on the flow cell of the SA sensor chip. With PBS pH 7.4, 1 mM TCEP, 0.05% Tween20, 2% DMSO as the running buffer, candidate molecules targeting PSMA with five serial dilution concentrations were injected into the flow cell at 30 L / min, with a binding time of 90 s. The buffer flow was maintained for 1800 s of dissociation. The KD value of the interaction between the candidate small molecule and the PSMA protein was evaluated using Biacore 8K evaluation software 1.0 and a fitting model for 1:1 binding.

[0291] The results are shown in Table 3. The binding affinity of DB-01 was slightly higher than that of PSMA-617.

[0292] Table 3 Binding kinetics and affinity of candidate molecules to PSMA protein determined by Biacore8K

[0293] Molecular number protein KD(M) ka(1 / Ms) kd(1 / s) PSMA-617 hPSMA-biotin 4.83E-10 1.01E+05 4.88E-05 DB-01 hPSMA-biotin 1.20E-10 2.22E+05 2.65E-05

[0294] Effect Example 2 Micro-SPECT / CT Scanning Imaging Experiment of Mice Transplanted with Human Prostate Cancer Cell Line 22RV1

[0295] Human prostate cancer cell line 22RV1 cells were subcutaneously injected into the armpits of nude mice to establish a prostate cancer tumor model. About 3.7 MBq (100 μCi / 200 μL) of the invention was intravenously injected 177 Lu-labeled compounds [ 177 Lu]DB-01, SPECT / CT images were collected using U-SPECT+ / CT (MI Labs) at 3, 5, 24, 72, 120, and 168 hours after injection, and the obtained SPECT / CT image results were quantitatively analyzed.

[0296] Table 4 shows the 177 Micro-SPECT / CT quantitative analysis results of the uptake of Lu]DB-01 in various organs of mice at 3 hours, 5 hours, 24 hours, 72 hours, 120 hours and 168 hours after injection, expressed as quantitative % injected dose (ID) / g.

[0297] From Table 4, we can see that [177 Lu]DB-01 is rapidly excreted through the kidneys and bladder in the early stages after injection.

[0298] exist[ 177 In the case of Lu]DB-01, its uptake in the tumor increased with time. According to calculations, the tumor uptake at 180 minutes was 13.55±3.85%ID / g, and the tumor retention time was significantly prolonged. At 168 hours, the tumor uptake was still 7.85±1.16%ID / g. 177 The renal uptake of Lu]DB-01 compounds decreased from 5 hours onwards and was rapidly excreted from the body.

[0299] Table 4 177 Lu]DB-01 uptake in various organs of mice over time (%ID / g)

[0300] time Tumor (22Rv1) kidney bladder liver muscle 3hr 21.25±4.38 43.00±7.38 65.50±36.65 2.75±0.90 1.97±0.71 5hr 21.28±0.42 19.29±2.77 140.36±10.51 1.56±0.15 1.10±0.56 24hr 15.51±2.65 1.32±0.63 0.61±0.00 0.91±0.45 0.51±0.17 72hr 12.41±0.95 1.38±0.29 1.03±0.62 0.90±0.42 0.81±0.23 120hr 8.90±0.48 0.77±0.62 0.92±0.06 0.99±0.22 0.43±0.17 168hr 7.85±1.16 1.53±0.46 1.01±0.38 1.04±0.26 0.65±0.34

[0301] From Table 4 and Appendix Figure 1 and attached Figure 2 You can see 177 After entering the mouse body, Lu-DB-01 was rapidly distributed to various organs and targets. The main metabolic pathway was through urination. After 5 hours, the concentration of radiopharmaceuticals in the tumor reached a peak, and then gradually decreased. After 24 hours, in addition to the accumulation of radiopharmaceuticals in the tumor, the accumulation of radiopharmaceuticals in other organs was greatly reduced. After 7 days, there was still a considerable amount of radiopharmaceuticals in the tumor. The results showed that the compound disclosed in this patent application not only had better PSMA cell affinity than the reference compound PSMA-617, but also had better pharmacokinetics, and was a more potential molecule as a diagnosis and treatment nuclear medicine.

[0302] Effect Example 3 Using Biacore to test the affinity of DB compounds for PSMA protein

[0303] The Biacore 8K (Cytiva) instrument was used to detect ligand binding of PSMA protein (Sino biological). PSMA protein was captured on the SA chip. Before ligand immobilization (flow paths 1, 2, flow rate of 10 μL / min), PSMA protein was immobilized on flow path 2 with flow buffer (10 μg / ml, flow rate of 5 μL / min, injection time of 600 s), and the sensor surface was conditioned by three consecutive injections of 1 M NaCl in 50 mM NaOH. After each ligand injection, an additional wash with isopropanol in 1 M NaCl and 50 mM NaOH was included (flow paths 1, 2, flow rate of 10 μL / min, injection time of 60 s).

[0304] All compounds were dissolved in 100% dimethyl sulfoxide and diluted to 10 mM, and then diluted to the appropriate maximum concentration in the assay buffer (PBS, pH 7.4, 1 mM TCEP (tris-(2-hydroxyethyl)phosphine), 0.05% P20, 2% dimethyl sulfoxide). The analytes were run using the following conditions: assay temperature = 15 °C, assay step = all set to LMW kinetics; cycle type = single cycle (contact time 90 s, dissociation time 1800 s, flow rate 30 μL / min, flow cell 1, 2); flow cell detection = 2-1). Data evaluation was performed using Biacore Insight evaluation software, and the data fit the 1:1 binding model.

[0305] The Biacore results are shown in Table 5: pKD = -LogKD, where KD is the binding affinity of the compound for the PSMA protein measured by Biacore. It is expressed as KD(M) = Kd(1 / s) / Ka(1 / Ms). A represents pKD > 8; B represents 8 < pKD < 7; C represents 7 < pKD < 6; D represents pKD > 6;

[0306] Table 5 Biacore test results

[0307]

[0308]

[0309] Effect Example 4 177 Lu labeling and animal biodistribution experiment

[0310] Tissue distribution of PSMA-positive tumor-bearing mice: Tumor-bearing animals (purchased from Shanghai Alamo Medical Technology Co., Ltd.), 3x10 6 22rv1 cells (in 50% Matrigel, Corning) were subcutaneously inoculated into the right shoulder of (Balb / c Nude) mice about 6-9 weeks old. When the tumors grew to an appropriate size, 177 Lu-radiolabeled DB compound was injected into the tail vein of the mice (about 1.85 MBq - 3.7 MBq / mouse, specific activity: 22423.82 KBq / μg). After the animals were administered the drug, they were euthanized by carbon dioxide inhalation at 24 h. After euthanasia, the blood and organs (blood, liver, kidney, muscle, tumor) of the animals were collected.

[0311] Blood was collected from the abdominal aorta, and immediately 100 μL was quantified into a designated centrifuge tube (weighed). After the organs were collected, they were washed twice with deionized water and dried, placed in a pre-weighed test tube, weighed again, and the sample weight was calculated. The samples were measured on the day of collection. All blood samples and tissue samples were measured for radioactivity counts using a gamma counter, and the results are as Figure 3 、 Figure 4 , Figure 5 , Figure 6 , Figure 7 And as shown in Table 6.

[0312] After intravenous administration, 177 Lu-labeled DB compounds were rapidly distributed to various organs of tumor-bearing mice. The uptake in non-targeted organs was always low and was quickly metabolized out of the body. The clearance rate in the blood was fast and was mainly excreted through kidney metabolism. 24 hours after administration, the uptake of Lu-177-labeled compounds on tumors remained at a high level, especially DB-12 and DB-19, which had abnormally high uptake in PSMA-positive 22RV1 tumor cells.

[0313] Table 6 177 Biodistribution data of Lu-labeled compounds (n=2)

[0314]

[0315]

[0316] Effect Example 5 Competitive Binding

[0317] (1) Prepare a cell suspension of 22Rv1 cells in the logarithmic growth phase, adjust the cell density to 1×104 / mL, and inoculate 1 mL into a 24-well cell culture plate. Culture in a 37°C incubator overnight.

[0318] (2) Aspirate the cell culture medium, wash the cells once with PBS, and add 975 μL of culture medium without additives.

[0319] (3) Add 25 μL of a fixed concentration of radioactively labeled PSMA-617 ligand (final concentration in culture medium: 2 μCi / mL) and different concentrations (final concentration in culture 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 unlabeled blocking compounds DB-18, DB-19, DB-13, and E-3 to each well.

[0320] (4) Incubate on ice for 2 hours.

[0321] (5) Wash the cells three times with ice-cold PBS.

[0322] (6) The cells were lysed with 0.5 mL of 1 M sodium hydroxide, washed twice with 0.5 mL of PBS, and the sodium hydroxide (0.5 mL) and PBS (0.5 mL x 2) solutions were collected to measure the uptake counts.

[0323] The experimental results are shown in Figure 8 The results showed that the IC50 values ​​of compounds E-3, DB-13, DB-18 and DB-19 were 2.06nM, 1.83nM, 2.58nM and 0.93nM, respectively.

[0324] Effect Example 6 Cell Killing

[0325] A. Prepare a cell suspension of LNCaP cells in the logarithmic growth phase, adjust the cell density to 1×105 / ml, and inoculate 1 ml into a 24-well cell culture plate. Culture in a 37°C incubator for 24 hours.

[0326] B. Use serum-containing culture medium to prepare 177Lu-labeled compound (177Lu-E-3) with radioactivity concentrations of 5, 15, and 45 μCi / mL for use (unlabeled E-3 uses a precursor concentration corresponding to 45 μCi / mL).

[0327] C. The experimental group was added with 1 mL / well of the above 177Lu labeled compound solution, and the control group was added with 1 mL / well of serum-containing culture medium.

[0328] D. Place in an incubator and culture for 24 hours, then replace all culture medium with fresh one.

[0329] E. On the fifth day, the enhanced CCK8 kit was used to detect the proliferation of cells in each group.

[0330] The experimental results are shown in Fig. 9 The results showed that at the tested dose, both compounds and the reference compound had a killing effect on cells, and the effect was dose-dependent. At the equivalent dose of 45 μCi / mL, DB-13 had a stronger cell killing effect, reaching a cell killing rate of 67%.

[0331] Effect Example 7 Determination of cLogP

[0332] Take 3 EP tubes, add 0.1 ml of saturated n-octanol and 80 μl of ultrapure water to each, and then add 20 μl (about 1 Mbq) of Lu-177 labeled DB-13, DB-18 and DB-19 compound solutions respectively. After oscillation, centrifuge at room temperature (2000 r / min, 5 min, centrifugal radius 10 cm). Take 100 μl from the lipid layer and water layer of each tube, measure the radioactivity counts per minute of the two phases, calculate LogP, and take the average of the results.

[0333] The experimental results are shown in Fig.10 The experimental logP of DB-13, DB-18, DB-19 and E-3 were -1.96, -1.86, -1.79 and -3.46, respectively. The results showed that all the tested compounds had good water solubility.

[0334] Effect Example 8 PPB Testing

[0335] Take the marked 177 Lu-E-3, 177 Lu-DB-13, 177 Lu-DB-18 and 177 Lu-DB-19 compound (1μCi), add 50μL PBS to obtain 20μCi / mL reaction solution. Take 50μL 20μCi / mL reaction solution and add it to 200μL plasma, repeat 3 tubes, mix well and incubate at room temperature for 10min, then add the sample to the ultrafiltration tube, centrifuge at 13000rpm for 45min, then add 50μL physiological saline, continue centrifugation for 15min, and count the cannula and filtrate separately.

[0336] PPB = [(upper layer counts - background counts)] / (lower layer counts + upper layer counts - 2*background counts)] * 100%.

[0337] The experimental results are shown in Fig.11 Compared with the reference compounds, the synthesized compounds have a higher albumin binding rate than DB-13 (DB-13: 90.84%, E-3: 86.61%). The albumin binding rates of DB-18 and DB-19 are lower than that of the reference compounds, which are 70.51% and 59.07%, respectively.

[0338] Effect Example 9 Treatment

[0339] 6-8 weeks old (Balb / c Nude) mice were subcutaneously inoculated with 3x106 cells of 22rv1 (in 50% Matrigel, Corning) on ​​the right shoulder blade of the animal. When the tumor grew to the required size for the experiment, the animals were randomly assigned to 11 experimental groups according to the tumor volume, with 5 animals in each group. The animal weight and tumor size were measured. On the day of grouping, the control group (normal saline), group 1 ( 177 Lu-DB-13 0.15mCi / each), Group 2 ( 177 Lu-DB-13 0.45mCi / each), Group 3 ( 177 Lu-DB-130.05mCi / each), Group 4( 177 Lu-B-11 0.05mCi / each), Group 5( 177 Lu-DB-11 0.15mCi / each), Group 6 ( 177 Lu-DB-11 0.45mCi / each), Group 7( 177 Lu-PSMA-617 0.15mCi / each), Group 8 ( 177 Lu-PSMA-617 0.45mCi / each), Group 9 ( 177Lu-DB-19 0.05mCi / each), Group 10( 177 Lu-DB-19 0.15mCi / each), Group 11( 177 Lu-DB-19 0.45mCi / animal) was administered, and general health and appearance observations were performed every day after the start of the experiment. Animal weight and tumor size were measured before each sample collection time point. Any abnormal observations found during the entire study will need to be recorded in the original data.

[0340] The experimental results are shown in Fig.12 , Fig.13 As shown in Table 7, DB-19 exhibited excellent tumor growth inhibition effects on tumor cells at two different doses (0.15 mCi and 0.45 mCi) compared with PSMA-617. DB-11 had a certain tumor growth inhibition effect at a high dose (0.45 mCi) after administration, but its inhibition decreased over time, but it was still higher than that of PSMA-617.

[0341] Table 7

[0342]

Claims

1. A compound, characterized in that The compound is:

Images