Method for radiolabeling GRPR antagonist, and kit of the same

Abstract

To provide a method for radiolabeling GRPR antagonist such as NeoB, and a kit of the same.SOLUTION: A method for labeling a radioactive isotope, preferably a gastrin-releasing peptide receptor (GRPR) antagonist by 68Ga, 67Ga, or 64Cu includes: i. a step of providing a first vial including the GRPR antagonist in a dry form; ii. a step of adding solution of the radioactive isotope into the first vial to obtain solution of the GRPR antagonist having the radioactive isotope; iii. a step of mixing the solution obtained in ii with at least one buffer and a step of incubating the mixture for a period enough for obtaining the GRPR antagonist labeled by the radioactive isotope; and iv. optionally, a step of adjusting pH of the solution.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present disclosure provides methods for radiolabeling GRPR antagonists, such as NeoB, and methods therefor. Regarding these kits. [Background technology]

[0002] Bombesin is a member of the European frog Bombina b It was first isolated from the mammalian gastrin-releasing peptide (GRP) and Demonstrated to be similar to euromedin B (NMB): Erspamer, VD discovery,Isolation,and Characterization of Bombesin-like Peptides.Ann NY Acad S ci 547:3-9,1988;Jensen,RT;Battey,JF; Spindel,ER;Benya,RVInternational uni on of pharmacology.LXVIII.Mammalian bomb esin receptors: Nomenclature, distribution ,pharmacology,signaling,and functions in normal and disease states.Pharmacol.Rev .2008,60,1-42].

[0003] Gastrin-releasing peptide (GRP), a bombesin-like peptide growth factor, is a gastrointestinal hormone It stimulates a number of functions in the gastrointestinal and central nervous systems, including hormone release, smooth muscle cell contraction, and epithelial cell proliferation. Gastrin-releasing peptide regulates many functions. It is a potent inhibitor of physiological and neoplastic tissues. Gastrin-releasing peptide is a potent mitogen and is involved in growth dysregulation and carcinogenesis. obtain.

[0004] The effects of GRP were investigated using a G protein-coupled receptor originally isolated from a small cell lung cancer cell line. is mediated primarily through binding to its receptor, the GRP receptor (GRPR) Upregulation of the GRP / GRPR pathway has been shown to be associated with breast, prostate, uterine, and ovarian cancers. several cancers, including: colon cancer, pancreatic cancer, gastric cancer, lung (small cell and non-small cell) cancer, and squamous cell carcinoma of the head and neck It has been reported in several cancers and in various brain and nerve tumors.

[0005] GRPR is highly overexpressed in prostate cancer and is expressed in human prostate cancer cell lines and xenografts. Studies in xenograft models have demonstrated high affinity (nM level) and high tumor accumulation (%ID / g). Although both showed relative expression of GRPR across early to late progressive disease has not yet been fully elucidated [Waters, et al. 2003, Br J Cancer.Jun 2;88(11):1808-1816].

[0006] In colorectal patients, the presence of GRP and expression of GRPR are associated with a variety of disease states, including LN and metastatic lesions. In randomly selected colon cancer samples, as determined by immunohistochemistry Over 80% of the samples aberrantly expressed GRP or GRPR, and over 60% expressed GRP and GRPR expression was observed in the adjacent normal healthy epithelium, whereas both [Scopinaro F, et al. Cancer Biother Radiopharm 2002,17(3):327-335].

[0007] GRP is normally present in pulmonary neuroendocrine cells and is essential for the development and maturation of the lung. However, it has been implicated in growth dysregulation and carcinogenesis. GRP stimulation appears to be associated with the release of epidermal growth factor receptor (EGFR) ligands. This leads to increased proliferation, followed by activation of EGFR and downstream mitogen-activated protein kinases. Using non-small cell lung cancer (NSCLC) cell lines, both EGF and GRP activate the It was confirmed that EGFR or GRPR stimulated NSCLC proliferation and resulted in cell death. It was confirmed [Shariati F, et al. Nucl Med Commun 20 14,35(6):620-625].

[0008] In nuclear medicine, peptide receptor agonists have long been a key factor in the development and use of tracers. The theory behind the use of agonist-based constructs The rationale is that the receptor-radioligand complex allows for high accumulation of radioactivity within the target cells. In the case of radioactive metal-labeled peptides, agonist stimulation was observed. Efficient receptor-mediated endocytosis in response to endocytosis is essential for optimal imaging of malignant diseases. This results in high in vivo radioactivity accumulation in the targeted tissue, a critical requirement for However, a paradigm shift occurred with the development of receptor-selective peptide antagonists. The preferred agonists include significantly higher in vivo tumor accumulation compared to highly potent agonists. A further advantage of GRPR antagonists is that they In clinical practice, the use of antagonists is not expected to result in acute biological side effects. The use of steroids is safer and, from the current diagnostic perspective, is more effective than that of steroids for potential therapeutic purposes. Even considering the larger volume, the tracer dose is not that large [S toykow C,et al.Theragnostics 2016,6(10): 1641-1650].

[0009] In non-clinical models, 68 Ga]-NeoB and [ 177 Lu]-NeoB([6 8Ga]-NeoBOMB1 and [ 177 Lu]-NeoBOMB1) High levels of GRPR are expressed in breast, prostate, and gastrointestinal stromal tumors (GISTs). It showed high affinity and low internalization upon binding to specific receptors. The ability of radiolabeled peptides to target expressing tumors was demonstrated in vivo in animal models. This was confirmed in imaging and biodistribution studies [Dalm et al Journal of nuclear medicine 2017,Vol.58(2):293- 299;Kaloudi et al.Molecules,2017 Nov 11; 22(11);Paulmichl A et al.Cancer Biother Radiopharm,2016 Oct;31(8):302-310].

[0010] however, 68 Ga, 67 Ga, or 64 NeoB was labeled with Cu, which This resulted in the preparation of a labeled NeoB solution for diagnostic imaging of GRPR-positive tumors in human patients. No optimized method for this purpose has been developed, especially in human subjects in need thereof. For intravenous injection in 68 Highly radiochemically pure labeled G, such as [Ga]NeoB There is a need for a rapid, efficient, and safe procedure that would provide an RPR antagonist. will be done. Summary of the Invention [Means for solving the problem]

[0011] A first aspect of the present disclosure is a method for producing a radioisotope, preferably 68 Ga, 67 Ga, or 64 C A method for labeling gastrin-releasing peptide receptor (GRPR) antagonists with u It is a law, i. providing a first vial containing said GRPR antagonist in a dried form. P, ii. adding the radioisotope solution into the first vial, thereby obtaining a solution of said GRPR antagonist with a radioisotope; iii. Mixing the solution obtained in ii. with at least one buffer, and and incubating it for a period of time sufficient to obtain the GRPR antagonist labeled with the isotopic structure. the step of substituting, iv. Optionally, adjusting the pH of the solution The present invention relates to a method comprising:

[0012] In certain embodiments, the radioisotope is 68 Ga, and in HPLC The radiochemical purity as determined by the method is at least 90%, and optionally, free 68 The percentage of Ga3+ (by HPLC) is less than 2% and / or uncomplexed body formation 68 The percentage of Ga3+ species (in the ITLC) is less than 5%.

[0013] In other specific embodiments, the radioisotope is: 67 Ga, and HPLC The radiochemical purity as measured in is at least 90%, and optionally, free 67 The percentage of Ga3+ (by HPLC) is less than 2% and / or non- complex formation 67 The percentage of Ga3+ species (in the ITLC) is less than 5%.

[0014] In other specific embodiments, the radioisotope is: 64 Cu, and HPLC The radiochemical purity as measured in is at least 90%, and optionally, free 64 The percentage of Cu2+ (by HPLC) is less than 2% and / or non- complex formation 64 The percentage of Cu2+ species (in ITLC) is below 5%.

[0015] Preferably, the GRPR antagonist is a NeoB compound of formula (I): [ka] (DOTA-(p-aminobenzylamine-diglycolic acid))-[D-Phe-Gln -Trp-Ala-Val-Gly-His-NH-CH[CH2-CH(CH3)2] 2.

[0016] In another aspect, the present disclosure provides a method for diagnosing tumors by diagnostic imaging in a subject in need thereof. by the methods disclosed herein for use as injections for in vivo detection. A radioisotope-labeled GRPR antagonist that can be obtained or is obtained by The present invention relates to a solution containing

[0017] Consists of the following ingredients in dry form: i. The following formula: CSP (where, C is a chelating agent capable of chelating said radioisotope; S is an optional spacer covalently linked between C and the N-terminus of P; P preferably has the general formula: Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Z (Xaa1 is absent or is selected from the amino acid residues Asn, Thr, Phe, 3-(2-thienoyl) 4-chlorophenylalanine (Cpa), α-naphthylalanine (α-phenylalanine), α-Naphthylalanine (α-Nal), β-naphthylalanine (β-Nal), 1,2,3,4-tetrahydrofuran Dolonorharman-3-carboxylic acid (Tpi), Tyr, 3-iodo-tyrosine (oI -Tyr), Trp, and pentafluorophenylalanine (5-F-Phe) (all L- or D-isomers); Xaa2 is Gln, Asn, or His; Xaa3 is Trp or 1,2,3,4-tetrahydronorharman-3-carboxylic acid (Tpi); Xaa4 is Ala, Ser, or Val; Xaa5 is Val, Ser, or Thr; Xaa6 is Gly, sarcosine (Sar), D-Ala, or β-Ala; Xaa7 is His or (3-methyl)histidine (3-Me)His; Z is -NHOH, -NHNH2, -NH-alkyl, -N(alkyl)2, and -O- alkyl, or Z is selected from [ka] where X is NH (amide) or O (ester), and R1 and R2 may be the same or different and may be selected from the group consisting of a proton, an optionally substituted alkyl group, alkyl, optionally substituted alkyl ether, aryl, aryl ether, or aryl alkyl, halogen, hydroxyl, hydroxyalkyl, amine, amino, amide do), or amide substituted aryl or heteroaryl groups a GRPR antagonist, which is a GRPR peptide antagonist; ii. Radiolytic protectants, such as gentisic acid; iii. bulking agents, such as mannitol; and iv. Optionally, a surfactant, such as macrogol 15 hydroxystearic acid It is another object of the present disclosure to provide a powder for injection solution comprising:

[0018] Typically, the powder for injection solution comprises the following components: i. NeoB of formula (I) in an amount between 20 and 60 μg, typically 50 μg; [ka] ii. gentisic acid in an amount of 50 and 250 μg, typically 200 μg; and iii. Mannitol in an amount between 10 and 30 mg, for example 20 mg; iv. Between 250 and 750 μg, e.g., 500 μg of macrogol 15-hydroxybenzoates Thearic acid.

[0019] The present disclosure provides a kit for carrying out the above labeling method, comprising: i. The following components in dry form: i. NeoB of the following formula (I): [ka] ii. Radiolytic protectants, such as gentisic acid; iii. optionally a bulking agent, such as mannitol, and iv. Optionally, a surfactant, such as macrogol 15 hydroxystearic acid, a first vial; and ii. a second vial containing at least one buffer, preferably in dry form; and to iii. Optionally, dissolve the radioisotopes generated by the radioisotope generator. Included cartridge for discharging The present invention further relates to a kit comprising:

[0020] Another kit disclosed herein comprises the following components in dry form: i. NeoB of the following formula (I): [ka] i. Radiolytic protectants, such as gentisic acid; ii. optionally a bulking agent, such as mannitol; iii. optionally a surfactant, such as macrogol 15 hydroxystearic acid; and iv. at least one buffer, preferably in dry form; and ii. Optionally, eluting radioisotopes generated by a radioisotope generator. Included cartridge for The vial contains a single vial having

[0021] For example, the kit may include the following components: i. NeoB of formula (I) in an amount between 20 and 60 μg, typically 50 μg; [ka] ii. gentisic acid in amounts of 50 and 250 μg, typically 200 μg; iii. Mannitol in an amount between 10 and 30 mg, for example 20 mg; iv. Optionally, between 250 and 750 μg, e.g., 500 μg of macrogol 15 Hydroxystearic acid The first or single vial may include DETAILED DESCRIPTION OF THE INVENTION

[0022] Generally, the present disclosure relates to the use of radioisotopes, preferably 68 Ga, 67 Ga, or 64 Cu A method for labeling gastrin-releasing peptide receptor (GRPR) antagonists. So, (i) providing a first vial containing said GRPR antagonist in a dried form; Top, (ii) adding the solution of the radioisotope into the first vial, thereby obtaining a solution of the GRPR antagonist having the radioisotope; (iii) mixing the solution obtained in ii. with at least one buffer; and incubating the GRPR antagonist for a period of time sufficient to obtain the isotopically labeled GRPR antagonist. incubating the mixture; and (iv) optionally adjusting the pH of the solution The present invention relates to a method comprising:

[0023] The radiolabeled GRPR antagonist obtained by the disclosed method is preferably P For use as a contrast agent for ET / CT, SPECT, or PET / MRI imaging It is a radioactive GRPR antagonist for

[0024] A preferred radiolabeled GRPR antagonist obtained by the disclosed method is a PET Suitable for use as a contrast agent for CT, SPECT, or PET / MRI imaging radioisotopes, preferably 68 Ga, 67 Ga, or 64 Ne labeled with Cu In a preferred embodiment, 67 Ga is for SPECT imaging diagnosis. Used for, and 68 Ga and 64 Cu is used for PET / CT or PET / MRI. Used for PET imaging.

[0025] The methods of the present disclosure advantageously produce radiolabeled compounds of excellent radiochemical purity, e.g., 68 G A radiolabeled NeoB compound having a .alpha. may be provided, typically as measured by HPLC. The radiochemical purity of the resulting product is at least 92%, and optionally, free 68 Ga3 The percentage of + (by HPLC) is less than 2% and / or uncomplexed 68 The percentage of Ga3+ species (in the ITLC) is less than 3%.

[0026] Radiochemical purity and free radicals were determined by HPLC or ITLC. 68 Measuring Ga3+ Assays for this purpose are further described in detail in the Examples.

[0027] definition The terms "treatment" and "treating" refer to the amelioration or arrest of a disease, disorder, or its symptoms. In particular, with respect to the treatment of tumors, the term "treatment" includes the inhibition of tumor growth or the reduction of tumor size. It may also refer to a decline in

[0028] Consistent with the International System of Units, "MBq" is an abbreviation for the unit of radioactivity "megabecquerel".

[0029] As used herein, "PET" stands for positron emission tomography.

[0030] As used herein, "SPECT" refers to single photon emission computed tomography (SPECT). Represents tomography.

[0031] As used herein, "MRI" stands for magnetic resonance imaging.

[0032] As used herein, "CT" stands for computed tomography.

[0033] As used herein, an "effective amount" or a "therapeutically effective amount" of a compound refers to a The term "antibody" refers to a compound that will elicit a biological or medical response in a subject, e.g., ameliorate a symptom. to alleviate the condition, slow or delay the progression of the disease, or prevent the disease. This refers to the amount of a compound that may be present.

[0034] As used herein, the terms "substituted" or "optionally substituted" " ranges from zero to the total number of open valences on the aromatic ring structure The term "substituted" refers to a group optionally substituted with one or more substituents selected from the following, in a number ranging from Halogen, -OR', -NR'R'', -SR', -SiR'R''R''', -O C(O)R', -C(O)R', -CO2R', -C(O)NR'R'', -OC(O) NR'R'', -NR''C(O)R', -NR'-C(O)NR''R''', -NR ''C(O)OR', -NR-C(NR'R''R''')=NR'''', -NR-C (NR'R'')=NR''' -S(O)R', -S(O)2R', -S(O)2NR 'R'', -NRSO2R', -CN, -NO2, -R', -N3, -CH(Ph)2, fluoro(C1-C4)alkoxo and fluoro(C1-C4)alkyl; and R' , R'', R''', and R'''' are hydrogen, alkyl, heteroalkyl, cycloalkyl, and independently selected from alkyl, heterocycloalkyl, aryl, and heteroaryl. When a compound of the present disclosure includes more than one R group, for example, each R group may be: When more than one R', R'', R''', and R'''' group is present, each Like these groups, they are independently selected.

[0035] As used herein, the term "alkyl" refers to any group, either alone or as part of another substituent. As a moiety, it refers to a linear or branched alkyl functional group having 1 to 12 carbon atoms. The alkyl groups are methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, s -butyl, and t-butyl, pentyl and its isomers (e.g., n-pentyl, iso-pentyl, hexyl), and hexyl and its isomers (e.g., n-hexyl, iso-hexyl). include.

[0036] As used herein, the term "heteroaryl" refers to a heteroaryl containing 5 to 10 atoms. having a single ring or multiple aromatic rings fused or covalently linked together; A polyunsaturated aromatic ring structure in which at least one ring is aromatic and at least one The ring atoms of are heteroatoms selected from N, O, and S. Nitrogen and sulfur heteroatoms may be optionally oxidized and the nitrogen heteroatom may be optionally quaternized Such rings may be fused to an aryl, cycloalkyl, or heterocyclyl ring. Non-limiting examples of such heteroaryls include: furanyl, thiophenyl, Pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, Isothiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, o Oxatriazolyl, thiatriazolyl zolyl), pyridinyl, pyrimidyl, pyrazinyl, pyridazinyl, oxazinyl, di Oxynyl (dioxinyl), tiadinyl, triazinyl, indolyl, isoindo aryl, benzofuranyl, isobenzofuranyl, benzothiophenyl, isobenzothiophene isobenzothiophenyl, indazolyl, benzimidazolyl, Benzoxazolyl, purinyl, benzothiadiazolyl lyl), quinolinyl, isoquinolinyl, cinnolinyl, quinazolinyl, and quinoxalyl Nil.

[0037] As used herein, the term "aryl" refers to an aryl group containing 6 to 10 ring atoms. Polyunsaturated aromatic hydrocarbons having a single ring or multiple aromatic rings fused together. At least one ring is aromatic. The aromatic ring may optionally be fused to and 1 to 2 additional rings (cycloalkyl, heterocyclyl, as defined herein) Suitable aryl groups include phenyl, naphthyl, , as well as benzopyranyl, benzodioxolyl, benzodioxanyl, and the like. This includes a phenyl ring fused to a heterocyclyl such as:

[0038] As used herein, the term "halogen" includes fluoro (-F), chloro (-F), (-Cl), bromo (-Br), or iodo (-I) groups.

[0039] As used herein, the term "optionally substituted fatty chain" refers to a fatty chain having a length of 4 to 3 an optionally substituted fatty chain having 6 carbon atoms, preferably 12 to 24 carbon atoms; Refers to...

[0040] As used herein, the term "chelator" refers to a compound that chelates a molecule through a non-covalent bond. It has a functional group such as an amine or carboxyl group suitable for forming a complex with a diisotopic isotope. Refers to the molecule.

[0041] As used herein, the term "radiolysis protector" refers to a substance that protects against, for example, radioactive Gamma rays emitted from nuclides break the bonds between atoms in organic molecules, forming radicals refers to a stabilizer that protects organic molecules from radiolysis when their radicals are then , are removed by a stabilizer, which removes the unwanted, possibly to prevent it from undergoing any other chemical reactions that may result in useless or even toxic molecules. Therefore, these stabilizers are also known as "free radical scavengers" or are also called "radical scavengers" for short. Other alternatives for these stabilizers The term "radiation stability enhancer," "radiolytic stabilizer," or simply "citric acid" is used interchangeably with "radiation stability enhancer," "radiolytic stabilizer," or simply "citric acid." "Char".

[0042] As used herein, the term "radiochemical purity" refers to a defined chemical or refers to the percentage of a given radionuclide present in biological form. radiochromatography, such as instant thin layer chromatography (iTLC) The radiochemical purity determination method is used in nuclear pharmacy. This is the most commonly accepted method for

[0043] Unless otherwise specified herein, "about" means ±20%, preferably ±10%, More preferably, it means ±5%, even more preferably ±2%, and even more preferably ±1%. The term "about" is used synonymously herein with "approximately."

[0044] providing a first vial containing said GRPR antagonist in dry form; i) GRPR antagonists As used herein, the GRPR antagonist has the following formula: CSP (where, C is a chelating agent capable of chelating a radioisotope; S is an optional spacer covalently linked between C and the N-terminus of P; P preferably has the general formula: Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Z (Xaa1 is absent or is selected from the amino acid residues Asn, Thr, Phe, 3-(2-thienoyl) 4-chlorophenylalanine (Cpa), α-naphthylalanine (α-phenylalanine), α-Naphthylalanine (α-Nal), β-naphthylalanine (β-Nal), 1,2,3,4-tetrahydrofuran Dolonorharman-3-carboxylic acid (Tpi), Tyr, 3-iodo-tyrosine (oI -Tyr), Trp, and pentafluorophenylalanine (5-F-Phe) (all L- or D-isomers); Xaa2 is Gln, Asn, or His; Xaa3 is Trp or 1,2,3,4-tetrahydronorharman-3-carboxylic acid (Tpi); Xaa4 is Ala, Ser, or Val; Xaa5 is Val, Ser, or Thr; Xaa6 is Gly, sarcosine (Sar), D-Ala, or β-Ala; Xaa7 is His or (3-methyl)histidine (3-Me)His; Z is -NHOH, -NHNH2, -NH-alkyl, -N(alkyl)2, and -O- alkyl, or Z is selected from [ka] where X is NH (amide) or O (ester), and R1 and R2 may be the same or different and may be selected from the group consisting of a proton, an optionally substituted alkyl group, alkyl, optionally substituted alkyl ether, aryl, aryl ether, or aryl alkyl, halogen, hydroxyl, hydroxyalkyl, amine, amino, amide do), or amide substituted aryl or heteroaryl groups It has a GRPR peptide antagonist).

[0045] According to one embodiment, Z is selected from one of the following formulae, where X is NH or O: is. [ka]

[0046] According to one embodiment, P is DPhe-Gln-Trp-Ala-Val-Gly-H is-Z; where Z is defined above.

[0047] According to one embodiment, P is DPhe-Gln-Trp-Ala-Val-Gly-H is-Z; Z is Leu-ψ(CHN)-Pro-NH and NH-CH(CH-CH(CH )2)2, or Z is selected from: [ka] where X is NH (amide) and R2 is CH(CH 2-CH(CH3)2, and R1 is the same as or different from R2 (CH2N)- It is Pro-NH2.

[0048] According to one embodiment, the chelating agent C is one chelating agent selected from the following list: It is obtained by grafting a silylating agent. [ka]

[0049] In certain embodiments, C is a chelating agent selected from the group consisting of: It is obtained by [ka]

[0050] According to one embodiment, S is selected from the group consisting of: a) an aryl containing residue of the formula: [ka] Here, PABA is p-aminobenzoic acid, PABZA is p-aminobenzyl alcohol, PDA is a phenylenediamine, and PAMBZA is an (aminomethyl) benzylamine; b) a dicarboxylic acid, ω-aminocarboxylic acid, ω-diaminocarboxylic acid, or dicarboxylic acid of the formula: Amine: [ka] where DIG is diglycolic acid and IDA is iminodiacetic acid; c) PEG spacers of various chain lengths, in particular PEG spacers selected from: [ka] d) α- and β-alpha chains of single or homogeneous chains of various chain lengths or heterogeneous chains of various chain lengths amino acids, especially [ka] GRP(1-18), GRP(14-18), GRP(13-18), BBN(l-5) , or [Tyr4]BB(1-5); or e) A combination of a, b, c, and d.

[0051] According to certain embodiments, the radiolabeled GRPR antagonist is selected from the group consisting of compounds of the formula: is selected from the group consisting of: [ka] wherein C and P are as defined above, and M is a radioisotope. Preferably, M is 68 Ga, 67 Ga, or 64 Cu.

[0052] According to one preferred embodiment, the GRPR antagonist is NeoB (NeoB) of formula (I): (also called oBOMB1): [ka] (DOTA-(p-aminobenzylamine-diglycolic acid))-[D-Phe-Gln -Trp-Ala-Val-Gly-His-NH-CH[CH2-CH(CH3)2] 2.

[0053] According to one embodiment, the radiolabeled GRPR antagonist is a radiolabeled N eoB2 is: [ka] (M-N4(p-aminobenzylamine-diglycolic acid)-[D-Phe-Gln-T rp-Ala-Val-Gly-His-NH-CH[CH2-CH(CH3)2]2; where M is a radionuclide.

[0054] According to another particular embodiment, the GRPR antagonist is Pro of formula (II): BOMB1 is: [ka] (DOTA-pABzA-DIG-D-Phe-Gln-Trp-Ala-Val-Gl y-His-Leu-ψ(CH2N)-Pro-NH2).

[0055] Synthesis of Compounds of Formulas (I), (II), and (III) The compounds of formula (I), (II) and (III) are described in the literature as "Positron Em ission Tomography Imaging of the Gastrin -Releasing Peptide Receptor with a Novel Bombesin Analogue”ACS Omega 2019,4,1470 It can be synthesized using the methods disclosed in US Pat. No. 6,478,478.

[0056] a first vial containing the GRPR antagonist; In some embodiments, the radiolabeling method uses a single vial kit. In one embodiment, the first vial contains the GRPR antagonist, both in dry form. Contains paste and buffer.

[0057] Instead, the radiolabeling method uses a two-vial kit. wherein the first vial contains the GRPR antagonist, and the second vial contains a buffer solution. Contains agents.

[0058] For example, the GRPR antagonist, typically a NeoB compound, is administered in an amount of 20 to 60 μg. An amount between 100 μg and 200 μg, typically 50 μg, is contained in the first vial.

[0059] The first vial optionally contains a radiolysis protectant, a bulking agent, and a surfactant ( Contains additional additives such as a tensioactive agent.

[0060] In a preferred embodiment, gentisic acid is preferably in an amount between 50 and 250 μg, Typically 200 μg may be used as a radiolysis protectant.

[0061] In a preferred embodiment, mannitol is used in an amount of, for example, between 10 and 30 mg, typically May be used as a bulking agent at 20 mg.

[0062] In a preferred embodiment, macrogol 15 hydroxystearic acid is, for example, 25 Amounts between 0 and 750 μg, typically 500 μg, may be used as surfactants. The surfactant advantageously prevents non-specific adhesion of the NeoB compound to glass or plastic surfaces. This reduced the specific adhesion, thereby optimizing the rate of the labeling process.

[0063] A preferred example of the first vial (vial 1 of the two-vial kit) is shown in Example 1. It is shown in

[0064] The first vial is preferably sterilized using methods well known in the art. Therefore, the first vial is obtained by freeze-drying or freeze-drying. It may also be provided in spray-dried form.

[0065] As used herein, a buffer is used in the incubation step (iii). , a buffer suitable for obtaining a pH between 3.0 and 6.0, preferably between 3.0 and 4.0 The "buffer for a pH of 3.0 to 6.0, preferably 3.0 to 4.0" is a useful Advantageously, it may be a formic acid buffer with sodium hydroxide.

[0066] The buffer may further comprise a first vial in embodiments using a single vial kit. in a separate second vial in embodiments using a two-vial kit It may be included.

[0067] (ii) adding the solution of the radioisotope into the first vial; Radioisotopes for use in radiolabeling methods include PET and SPE Suitable as contrast agents in CT imaging, including: 111 In, 133m In, 99m Tc, 94m Tc, 67 Ga, 66 Ga, 68 Ga, 52 Fe, 72 As, 97 Ru, 203 Pb, 62 Cu, 64 Cu, 86 Y, 51 Cr, 52m Mn, 157 Gd, 169 Yb, 172 Tm, 117m Sn, 89 Zr, 43 Sc , 44 Sc.

[0068] According to a preferred embodiment, the radioisotope is 68 Ga, 67 Ga, or 64 Cu In a preferred embodiment, 67 Ga is used for SPECT imaging, and 68 Ga and 64 Cu is used in PET imaging diagnostics such as PET / CT or PET / MRI. Used for:

[0069] The metal ions of such radioisotopes are attached to the functional groups of chelators, e.g., GRPR anta It forms a non-covalent bond with the carborboxylic acid of the agonist. It can be achieved.

[0070] In certain embodiments, the solution of the radioisotope comprises: i. A process for producing radioisotopes from parent non-radioactive elements using a radioisotope generator. Tep, ii. The radioisotope is eluted into HCl as an elution solvent. Separating the element from the iii. collecting the eluate; thereby obtaining a solution of said radioisotope in HCl. The eluate obtained from

[0071] radioactive isotope 68 The solution containing Ga is i. The generator generates the parent element 68 From Ge 68 Producing Ga element Steps to take ii. Optionally, an element 68 Ge / 68 By passing Ga through a suitable cartridge And it was generated 68 Ga element 68 Separated from Ge element, 68 Ga was dissolved in HCl , thereby obtaining a solution of said radioisotope in HCl. This is the eluate typically obtained from

[0072] 68 Ge / 68 From Ga generator 68 Such a method for producing Ga includes: It is well known in the art, for example Martiniova L, et al. Gallium-68 in Medical Imaging.Curr Radio pharm.2016;9(3):187-20;Dash A,Chakravart y Radionuclide generators:the prospect o f availing PET radiotracers to meet curr ent clinical needs and future research d emands R Am J Nucl Med Mol Imaging.2019 Feb 15;9(1):30-66.

[0073] radioactive isotope 68 The solution containing Ga is preferably obtained from cyclotron production. Such production may be carried out as described, for example, in Am J Nucl Med In Mol Imaging 2014;4(4):303-310, or BJ B. Nelson et al. / Nuclear Medicine and Bio This is described in Journal of Clinical Chemistry 80-81(2020)24-31.

[0074] Preferably, 68 Ga is preferably ionized by a cyclotron at 8 to 18 MeV. and more preferably between 11 and 14 MeV. It may be generated. 68 Ga uses a solid or liquid target system 68 Zn(p ,n) 68 The target may be produced via a Ga reaction. 68 Zn metal or 6 8 After irradiation, the target is transferred to further chemical treatment, 68 Ga is isolated using ion exchange chromatography. 68 Ga in HCl solution It is eluted.

[0075] Instead, the radioisotope is 67 The impact particles are protons and de Zinc (enriched or natural) with euterons, alpha particles, or helium(III) Or use a copper or germanium target 67 Various methods for the production of Ga include H elus, F., Maier-Borst, W., 1973. A comparativ. e investigation of methods used to produce ce 67Ga with a cyclotron.In:Radiopharmac euticals and Labeled Compounds,Vol.1,IA EA,Vienna,pp.317-324, ML Thakur Gallium- 67 and indium-111 radiopharmaceuticals I nt.J.Appl.Rad.Isot.,28(1977),pp.183-201, & Bjornstad, T., Holtebekk, T., 1993.Product ion of 67 Ga at Oslo cyclotron.University of Oslo Report OUP8-3-1, pp.3-5 It has been reported that protons of moderate energy (up to 64 MeV) nat The impact of Ge targets was also investigated by T Horiguchi, H Kumahora, and HI noue, Y Yoshizawa Excitation functions of Ge(p,xnyp)reactions and production of 6 8Ge,Int.J.Appl.Radiat.Isot.,34(1983),pp. 1531-1535, which is a suitable method for producing 67Ga. do.

[0076] Preferably, 67Ga may be produced by a cyclotron. 68 Zn(p, 2n) 67 From Ga 67 Such methods for producing Ga are well known in the art. It is well known, for example, Alirezapour B et al. Iranian J ournal of Pharmaceutical Research(2013), 12(2):355-366. More preferably, the method comprises the steps of: It uses proton beams with energies between 40 and 40 MeV. 67 Ga is a solid or liquid target Using the system 67 Zn(p,n) 67 Ga or 68 Zn(p,2n) 67 Ga reaction The target may be produced through either enrichment or 67 Zn or 68 Zn metal mater consisted of a liquid solution. After irradiation, the target was transferred to further chemical treatment, 67 Ga It is isolated using ion exchange chromatography. Upon final evaporation from aqueous HCl twist 67 GaCl3 is produced, which is then added to the single vial for labeling procedures. may be added.

[0077] Instead, the radioisotope is obtained from cyclotron production. 64 Cu Such a production method is described, for example, in WO 2013 / 029616. It is described in the

[0078] Typically, 64 Cu is preferably ionized by a cyclotron at between 11 and 18 MeV. The proton beam may be produced using a proton beam with an energy of .mu.m. 64 Cu is a solid or liquid target. Using the system, 64 Ni(p,n) 64 It may also be produced via a Cu reaction. -The target is 64 Ni metal or 64 After irradiation, the target was further The waste is then subjected to a chemical treatment 64 Cu was isolated using ion exchange chromatography. The final evaporation of the HCl solution 64 CuCl2 is produced, which is then labeled may be added to the first vial for the method.

[0079] The solution obtained in step (ii) is mixed with at least one buffer, and the radioactive and incubating the GRPR antagonist for a period of time sufficient to obtain the isotopically labeled GRPR antagonist. Incubating step (iii) The radiolabel is added to the first vial containing the GRPR antagonist (e.g., NeoB compound). with a radioisotope (typically one of the 68 Ga, 67 Ga, or 64 Cu) The method begins after mixing the solution containing the compound in an appropriate buffer as disclosed above.

[0080] In certain embodiments, the incubating step is performed at a temperature between 80°C and 100°C. It is preferably carried out at a temperature between 90°C and 100°C, typically around 95°C.

[0081] In certain embodiments, the incubating step is for 5 to 10 minutes, e.g. It lasts between 6 and 8 minutes, typically running for about 7 minutes.

[0082] At the end of the labeling process, the radioisotope (68Ga, 67 Ga, or 64Cu, etc.) A sequestering agent with specific affinity for the isotope chelates the unreacted portion of the isotope. The sequestering agent and the unreacted radioisotope may be added to This complex can then be discarded to increase radiochemical purity after radiolabeling. good.

[0083] 68 Preferred embodiments of the method for radiolabeling NeoB with Ga The present disclosure more particularly relates to: 68 For labeling NeoB compounds of formula (I) with Ga 1. A method comprising: [ka] (DOTA-(p-aminobenzylamine-diglycolic acid))-[D-Phe-Gln -Trp-Ala-Val-Gly-His-NH-CH[CH2-CH(CH3)2] 2; i. A solution containing approximately 50 μg of NeoB and between 50 and 250 μg of gentisic acid in a dry form. providing a first vial having ii. Adding HCl to the first vial 68 adding a solution of Ga; iii. The solution obtained in ii. is adjusted to a pH in the range of 3.0 to 4.0. is mixed with a buffer, 68 for a period of time sufficient to obtain the Ga-labeled NeoB compound. , incubating it; iv. Optionally, adjusting the pH of the solution The present invention relates to a method comprising:

[0084] In certain embodiments of the method,68 The solution of Ga i. The generator generates the parent element 68 From Ge 68 Producing Ga element; ii. Optionally, an element 68 Ga / 68 By passing Ge through a suitable cartridge And it was generated 68 Ga element 68 Separated from Ge element, 68 Ga was dissolved in HCl , thereby obtaining a solution of said radioisotope in HCl. The eluate obtained from

[0085] Typically, the buffer consists of 60 mg of formic acid and 56.5 mg of sodium hydroxide. do.

[0086] Advantageously, in certain embodiments, simple labeling of the GRPR antagonist is achieved by measuring the eluate is commercially available without any treatment or any additional purification steps. 68 G e / 68 in HCl produced by Ga generator 68 Even if it is obtained by elution of Ga good.

[0087] Powder for injection solution The present disclosure provides a composition comprising the following components in dry form: i. a GRPR antagonist as defined above, typically of formula (I) as defined above; NeoB; ii. Radiolytic protectants, such as gentisic acid; iii. bulking agents, such as mannitol; and iv. Optionally, a surfactant, such as macrogol 15 hydroxystearic acid The present invention further relates to a powder for injection solution comprising

[0088] A preferred embodiment comprises the following components: i. NeoB of formula (I) in an amount between 20 and 60 μg, typically 50 μg; [ka] ii. gentisic acid in an amount between 50 and 250 μg, typically 200 μg; and iii. Mannitol in an amount between 10 and 30 mg, for example 20 mg; iv. Between 250 and 750 μg, e.g., 500 μg of macrogol 15-hydroxybenzoates stearic acid Includes:

[0089] Radiolabeling Kits of the Present Disclosure The present disclosure also provides a kit for carrying out the above labeling method, comprising: i. A first vial having the following components in dry form: i. a GRPR antagonist as defined above, ii. Radiolytic protectants, such as gentisic acid; iii. optionally a bulking agent, such as mannitol, and iv. optionally a surfactant, such as macrogol 15 hydroxystearic acid; and ii. a second vial containing at least one buffer, preferably in dry form; and to iii. Optionally, dissolve the radioisotopes generated by the radioisotope generator. Included cartridge for discharging The present invention also relates to a kit comprising:

[0090] Preferably, said first or single vial contains the following components: i. NeoB of formula (I) in an amount between 20 and 60 μg, typically 50 μg; [ka] ii. gentisic acid in an amount between 50 and 250 μg, typically 200 μg; iii. Mannitol in an amount between 10 and 30 mg, for example 20 mg; and iv. Optionally, between 250 and 750 μg, e.g., 500 μg of macrogol 15 Hydroxystearic acid Includes:

[0091] The second vial or the single vial may be filled with a solution containing 100% ethanol to maintain a pH between 3.0 and 4.0. For example, the second vial may contain a buffer containing formic acid and water as a buffer. Contains sodium oxide.

[0092] Preferably, all components of the first, second, or single vial are in dry form. He is acting like this.

[0093] The radioisotopes used to label GRPR antagonists are available as ready-to-use products. That is, mix with the first vial and buffer provided in the kit and incubate. Alternatively, the radiation may be provided with the kit to specifically Sex isotopes are 68 Ga, 67 Ga, and 64 Cu, which has a relatively short half-life If so, mix with the first vial and buffer and release immediately before incubating. It may be eluted from a radioisotope generator.

[0094] Preferably, the components are packaged together with instructions for carrying out the methods according to the present disclosure. The product is placed in a sealed container which may be emptied.

[0095] The kit also includes a gallium-69 generator for dissolution and / or subsequent mixing and heating. Performed automatically, used as part of an automated system or remotely controlled mechanism In this embodiment, the vial containing the GRPR antagonist (the Vial 1) is directly connected to the elution system and / or heating system.

[0096] The kit may be particularly adapted for use in the methods disclosed in the next section. stomach.

[0097] In a specific embodiment, the GRPR antagonist is NeoB as defined above. do.

[0098] Use of kits according to the present disclosure The kit as defined above is particularly suitable for use in the labeling method disclosed in the previous paragraph. may be applied.

[0099] Advantageously, radioisotopes (e.g. 68 Ga, 67 Ga, or 64 Cu) The solution containing the GRPR antagonist (e.g., NeoB compound) can be prepared as described in the previous section. can be obtained or obtained by the labeling method described above.

[0100] Such solutions can be used, for example, to detect tumors by diagnostic imaging in a subject in need thereof. It may also be ready for use as an injection for in vivo detection.

[0101] In some embodiments, the subject is a mammal, e.g., a rodent, canine, or feline. In a preferred embodiment, the subject is a mammal, including, but not limited to, a human, a mammal, or a primate. , human.

[0102] The need for an effective pharmaceutical carrier for an injectable composition is well known to those skilled in the art ( For example, Pharmaceutics and Pharmacy Practice, J .B. Lippincott Company, Philadelphia, PA, Ba nker and Chalmers, eds., pages 238-250(198 2), and ^SHP Handbook on Injectable Drugs,T See Rissel, 15th ed., pages 622-630 (2009). sea bream).

[0103] Typically, the solution for use as an injection is administered to a subject in need thereof. To achieve this, a single dose of between 150 and 250 MBq of [68Ga]-NeoB is provided.

[0104] In certain embodiments, the subject in need thereof is a subject with cancer. In particular, prostate cancer, breast cancer, small cell lung cancer, colon cancer, gastrointestinal stromal tumors, and gastrinomas , glioma, glioblastoma, renal cell carcinoma, gastroenteropancreatic neuroendocrine tumor, esophageal squamous cell tumor, neuro Blastoma, head and neck squamous cell carcinoma, and other neoplasm-related tumors that may be GRPR-positive Patients with tumors selected from ovarian tumors, endometrial tumors, and pancreatic tumors that exhibit vasculature. do.

[0105] Typically, PET / MRI, SPECT, or PET / CT imaging involves the application of radiation to a subject. Preferably, the subject is administered the labeled GRPR antagonist within 1 to 4 hours after the administration. It may be performed 2 and 3 hours after administration of the labeled GRPR antagonist.

[0106] Embodiment The following specific embodiments are disclosed: 1. Radioisotopes, preferably 68 Ga, 67 Ga, or 64 Cu induces gastrin release 1. A method for labeling a peptide receptor (GRPR) antagonist, comprising: i. providing a first vial containing said GRPR antagonist in a dried form. P, ii. adding the radioisotope solution into the first vial, thereby obtaining a solution of said GRPR antagonist with a radioisotope; iii. Mixing the solution obtained in ii. with at least one buffer, and and incubating it for a period of time sufficient to obtain the GRPR antagonist labeled with the isotopic structure. the step of substituting, iv. Optionally, adjusting the pH of the solution A method comprising: 2. The first vial in step i. contains the GRPR, preferably both in dry form. 2. The method of embodiment 1, wherein the reaction vial comprises the antagonist and a buffer. 3. Step iii: Adding the solution obtained in step ii. to at least one solution containing a buffer. mixing the radioisotope-labeled GRPR antagoniza- tion with a reaction solution; 2. The method of embodiment 1, comprising incubating the same for a period of time sufficient to obtain a stent. . 4. Any of embodiments 1 to 3, wherein the solution having the radioisotope further comprises HCl. There is one way. 5. The radioisotope is 68 Ga and radiochemically determined by HPLC The purity is at least 92% and, optionally, free 68 Percentage of Ga3+ Di (by HPLC) is less than 2% and / or uncomplexed 68 Ga3+ species Any one of embodiments 1 to 4, wherein the percentage (in ITLC) is 3% or less. Two ways. 6. The radioisotope is 67 Ga and radiochemically determined by HPLC The purity is at least 92% and, optionally, free 67 Percentage of Ga3+ Di (by HPLC) is less than 2% and / or uncomplexed 67 Ga3+ species Any one of embodiments 1 to 4, wherein the percentage (in ITLC) is 3% or less. Two ways. 7. The radioisotope is 64 Cu and radiochemically determined by HPLC The purity is at least 92% and, optionally, free 64 Percentage of Cu2+ Di (by HPLC) is less than 2% and / or uncomplexed 64 Cu2+ species Any one of embodiments 1 to 4, wherein the percentage (in ITLC) is 3% or less. Two ways. 8. The GRPR antagonist has the following formula: CSP (where, C is a chelating agent capable of chelating said radioisotope; S is an optional spacer covalently linked between C and the N-terminus of P; P preferably has the general formula: Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Z; (Xaa1 is absent or is selected from the amino acid residues Asn, Thr, Phe, 3-(2-thienoyl) 4-chlorophenylalanine (Cpa), α-naphthylalanine (α-phenylalanine), α-Naphthylalanine (α-Nal), β-naphthylalanine (β-Nal), 1,2,3,4-tetrahydrofuran Dolonorharman-3-carboxylic acid (Tpi), Tyr, 3-iodo-tyrosine (oI -Tyr), Trp, and pentafluorophenylalanine (5-F-Phe) (all L- or D-isomers); Xaa2 is Gln, Asn, or His; Xaa3 is Trp or 1,2,3,4-tetrahydronorharman-3-carboxylic acid (Tpi); Xaa4 is Ala, Ser, or Val; Xaa5 is Val, Ser, or Thr; Xaa6 is Gly, sarcosine (Sar), D-Ala, or β-Ala; Xaa7 is His or (3-methyl)histidine (3-Me)His; Z is -NHOH, -NHNH2, -NH-alkyl, -N(alkyl)2, and -O- alkyl, or Z is selected from [ka] where X is NH (amide) or O (ester), and R1 and R2 may be the same or different and may be selected from the group consisting of a proton, an optionally substituted alkyl group, alkyl, optionally substituted alkyl ether, aryl, aryl ether, or aryl alkyl, halogen, hydroxyl, hydroxyalkyl, amine, amino, amide do), or amide substituted aryl or heteroaryl groups 8. The compound of any one of embodiments 1 to 7, wherein the compound is a GRPR peptide antagonist of any one of the methods; and 9.P is DPhe-Gln-Trp-Ala-Val-Gly-His-NH-CH( The method of embodiment 8, wherein the carboxyl group is CH2-CH(CH3)2). 10. The GRPR antagonist has the formula (I): [ka] DOTA-(p-aminobenzylamine-diglycolic acid)-D-Phe-Gln-T rp-Ala-Val-Gly-His-NH-CH[CH2-CH(CH3)2]2. 9. The method of embodiment 8, wherein the NeoB compound is 11. The GRPR antagonist is administered in an amount between 20 and 60 μg, typically 50 μg; 11. The method of any one of embodiments 1-10, contained in said first vial. 12. The first vial preferably contains between 50 and 250 μg, typically 200 μg. 12. Any of embodiments 1 to 11, further comprising gentisic acid as a radiolysis protectant. Or one way. 13. The first vial contains, for example, between 10 and 30 mg, typically 20 mg, and 13. The method of any one of embodiments 1-12, further comprising mannitol as a bulking agent. 14. The first vial contains, for example, between 250 and 750 μg, typically 500 μg and further comprising macrogol 15 hydroxystearic acid as a surfactant. One of the methods 1 to 13. 15. The buffer has a pH between 3.0 and 4.0 in incubating step (iii). 15. The method of any one of embodiments 1-14, wherein H is present in an amount suitable to provide 16. Any of embodiments 1-15, wherein the buffer comprises formic acid and sodium hydroxide. One way. 17. The incubating step is carried out at a temperature between 80°C and 100°C, preferably between 90°C and 1 17. Any one of embodiments 1-16, wherein the reaction is carried out at a temperature between 0°C and 100°C, typically at about 95°C. How to do it. 18. The incubation step should last between 5 and 10 minutes, e.g., between 6 and 8 minutes. 18. The method of any one of embodiments 1-17, wherein the method is performed for a period of time, typically about 7 minutes. 19. The solution of the radioisotope i. A process for producing radioisotopes from parent non-radioactive elements using a radioisotope generator. Tep, ii. The radioisotope is eluted into HCl as an elution solvent. Separating the element from the iii. collecting the eluate; thereby obtaining a solution of said radioisotope in HCl. 19. The method of any one of embodiments 1 to 18, wherein the eluate is obtained from 20. 68 1. A method for labeling a NeoB compound of formula (I) with Ga, comprising: [ka] (DOTA-(p-aminobenzylamine-diglycolic acid))-D-Phe-Gln- Trp-Ala-Val-Gly-His-NH-CH[CH2-CH(CH3)2]2 i. A solution containing approximately 50 μg of NeoB and between 50 and 250 μg of gentisic acid in a dry form. providing a first vial having ii. Adding HCl to the first vial 68 adding a solution of Ga; iii. The solution obtained in ii. is adjusted to a pH in the range of 3.0 to 4.0. is mixed with a buffer, 68 for a period of time sufficient to obtain the Ga-labeled NeoB compound. , incubating it; iv. Optionally, adjusting the pH of the solution A method comprising: 21. The above in HCl 68 The solution of Ga i. The generator generates the parent element 68 From Ge 68 Producing Ga element; ii. Optionally, an element 68 Ga / 68 By passing Ge through a suitable cartridge And it was generated 68 Ga element 68 Separated from Ge element, 68 Ga was dissolved in HCl , thereby obtaining a solution of said radioisotope in HCl. 21. The method of embodiment 20, wherein the eluate is obtained from 22. The buffer consists of 60 mg of formic acid and 56.5 mg of sodium hydroxide. 22. The method of embodiment 20 or 21. 23. The incubating step is carried out at a temperature between 80°C and 100°C, preferably between 90°C and 1 23. Any one of embodiments 20-22, wherein the reaction is carried out at a temperature between 0°C and 100°C, typically at about 95°C. Two ways. 24. The incubation step should last between 5 and 10 minutes, e.g., between 6 and 8 minutes. 24. The method of any one of embodiments 20 to 23, wherein the method is performed for a period of time, typically about 7 minutes. 25. Injection for in vivo detection of tumors by imaging in a subject in need thereof. 25. The method of claim 1, wherein the liquid is obtained by the method of any one of embodiments 1 to 24. A solution containing a radioisotope-labeled GRPR antagonist that can be obtained or is capable of being administered. 26. Injection for in vivo detection of tumors by diagnostic imaging in a subject in need thereof. 25. The method of claim 20, wherein the composition is obtained by the method of any one of embodiments 20 to 24. be able to or obtain 68 A solution containing Ga-labeled NeoB compounds. 27. The tumor is selected from tumors expressing GRPR, preferably the GR PR-expressing tumors include prostate cancer, breast cancer, small cell lung cancer, colon cancer, and gastrointestinal stromal tumors. , gastrinoma, renal cell carcinoma, gastroenteropancreatic neuroendocrine tumor, esophageal squamous cell tumor, neuroblastoma cytoma, head and neck squamous cell carcinoma, and oocytes showing GRPR-positive neoplasia-associated vasculature 27. The method according to claim 25, wherein the tumor is selected from a pancreatic tumor, an endometrial tumor, and a pancreatic tumor. Solution for use. 28. A dry form of the following components: i. The following formula: CSP (where, C is a chelating agent capable of chelating said radioisotope; S is an optional spacer covalently linked between C and the N-terminus of P; P preferably has the general formula: Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Z; (Xaa1 is absent or is selected from the amino acid residues Asn, Thr, Phe, 3-(2-thienoyl) 4-chlorophenylalanine (Cpa), α-naphthylalanine (α-phenylalanine), α-Naphthylalanine (α-Nal), β-naphthylalanine (β-Nal), 1,2,3,4-tetrahydrofuran Dolonorharman-3-carboxylic acid (Tpi), Tyr, 3-iodo-tyrosine (oI -Tyr), Trp, and pentafluorophenylalanine (5-F-Phe) (all L- or D-isomers); Xaa2 is Gln, Asn, or His; Xaa3 is Trp or 1,2,3,4-tetrahydronorharman-3-carboxylic acid (Tpi); Xaa4 is Ala, Ser, or Val; Xaa5 is Val, Ser, or Thr; Xaa6 is Gly, sarcosine (Sar), D-Ala, or β-Ala; Xaa7 is His or (3-methyl)histidine (3-Me)His; Z is -NHOH, -NHNH2, -NH-alkyl, -N(alkyl)2, and -O- alkyl, or Z is selected from [ka] where X is NH (amide) or O (ester), and R1 and R2 may be the same or different and may be selected from the group consisting of a proton, an optionally substituted alkyl group, alkyl, optionally substituted alkyl ether, aryl, aryl ether, or aryl alkyl, halogen, hydroxyl, hydroxyalkyl, amine, amino, amide do), or amide substituted aryl or heteroaryl groups a GRPR antagonist, which is a GRPR peptide antagonist; ii. Radiolytic protectants, such as gentisic acid; iii. bulking agents, such as mannitol; and iv. Optionally, a surfactant, such as macrogol 15 hydroxystearic acid 1. A powder for injection solution, comprising: 29. The GRPR antagonist is a NeoB compound of formula (I): Powder for injection solution according to form 28. [ka] 30. The NeoB compound is included in an amount between 20 and 60 μg, typically 50 μg. Powder for injection solution according to Form 29. 31. Gentisic acid is present in amounts between 50 and 250 μg, typically 200 μg. The powder for injection solution according to any one of embodiments 28 to 30. 32. The bulking agent is mannitol in an amount between 10 and 30 mg, for example, 20 mg. A powder for injection solution according to any one of embodiments 28 to 31. 33. The surfactant is present in an amount between 250 and 750 μg, for example 500 μg of macrogol. For the injectable solution of any one of embodiments 28 to 32, wherein the compound is 15-hydroxystearic acid. Powder. 34. Constituents of: - an amount between 20 and 60 μg, typically 50 μg, of NeoB of formula (I) below; [ka] - an amount between 50 and 250 μg, typically 200 μg of gentisic acid, and - an amount between 10 and 30 mg, for example 20 mg of mannitol, and -Amounts between 250 and 750 μg, e.g., 500 μg of macrogol 15 hydroxystearic acid phosphoric acid 34. The powder for injection solution of any one of embodiments 28 to 33, comprising: 35. i. The following components in dry form: NeoB of formula (I): [ka] radioprotectants, e.g. gentisic acid, optionally a bulking agent, such as mannitol, and - optionally a surfactant, such as macrogol 15 hydroxystearic acid a first vial having: ii. a second vial containing at least one buffer, preferably in dry form; and to iii. Optionally, dissolve the radioisotopes generated by the radioisotope generator. Included cartridge for discharging 21. A kit for carrying out the method of embodiment 20, comprising: 36. i. The following components in dry form: NeoB of formula (I): [ka] radioprotectants, e.g. gentisic acid, optionally a bulking agent, e.g., mannitol; optionally a surfactant, such as macrogol 15 hydroxystearic acid; and at least one buffer, preferably in dry form a single vial having ii. Optionally, eluting radioisotopes generated by a radioisotope generator. Included cartridge for 21. A kit for carrying out the method of embodiment 20, comprising: 37. The NeoB compound is included in an amount between 20 and 60 μg, typically 50 μg. 37. The kit of claim 35 or 36. 38. Gentisic acid is present in amounts between 50 and 250 μg, typically 200 μg. The kit according to any one of embodiments 35 to 37. 39. The bulking agent is mannitol in an amount between 10 and 30 mg, for example, 20 mg. The kit of any one of embodiments 35 to 38. 40. The surfactant is present in an amount between 250 and 750 μg, for example 500 μg of macrogol. 40. The kit of any one of embodiments 35 to 39, wherein the hydroxystearic acid is 15-hydroxystearic acid. 41. The first or single vial contains the following components: - an amount between 20 and 60 μg, typically 50 μg, of NeoB of formula (I) below; [ka] - Amounts between 50 and 250 μg, typically 200 μg of gentisic acid, - an amount between 10 and 30 mg, for example 20 mg of mannitol, and Optionally, in an amount between 250 and 750 μg, e.g., 500 μg of macrogol 15 hydrochloride Roxystearic Acid 41. The kit of any one of embodiments 35 to 40, comprising: 42. The second vial or single vial maintains a pH between 3.0 and 4.0. 42. The kit of any one of embodiments 35 to 41, comprising a buffer for: 43. An embodiment in which the second vial contains formic acid and sodium hydroxide as buffering agents. One of the kits 35-42. 44. All components of the first, second, or single vial are in dry form. 44. The kit of any one of embodiments 35 to 43, wherein [Example]

[0107] Hereinafter, the disclosure will be described in more detail, particularly with reference to examples, which are not intended to limit the invention. It is not intended to determine

[0108] Radiochemical purity by ITLC Preparation of mobile phase solutions: Ammonium acetate 5M: 3.85 g (3.84 mL) in a 10 mL graduated flask Accurately weigh 100g of ammonium acetate (615 ÷ 3.85385g) and add 100g of ammonium acetate. Dissolve in 0 mL of MilliQ water.

[0109] Ammonium acetate / MeOH: Using a graduated cylinder, add 1 mL of ammonium acetate. Add 2 mL of 5M solution, 2 mL of MilliQ water, and 7 mL of methanol. Transfer the eluent to the bar.

[0110] Preparation of ITLC-SA: Cut one ITLC-SA into 115 mm pieces for each vial. Then, draw a line 20 mm from the bottom (where to place a 5 uL sample droplet) and Draw a line 100 mm from the center (where chromatographic development should stop). . 68 Ga-NeoB: Reference factor 0.6~0.9 68 Ga uncomplexed species: Reference factor = 0.0 ÷ 0.1 ( 68 The Ga uncomplexed species is 68 Ga colloidal species and free form 68 (Refers to Ga).

[0111] Radiochemical purity by HPLC

[0112] [Table 1]

[0113] Example 1: Radiolabeling NeoB with 68Gallium Using a 2-Vial Kit Developing methods for 1.2-Vial Kit Description and Composition Applicant has developed a sterile 2-vial kit consisting of: Vial 1: 68 Ge / 68 Gallium chloride in HCl eluted from a Ga generator Mu-68( 68 NeoB, 50 μg, powder for injection solution, dissolved in a solution of GaCl3 End; Vial 2: Reaction buffer.

[0114] Add vial 2 to the dissolved vial 1.

[0115] One accessory cartridge is used to remove any germanises potentially present in the generator eluate. It is used to reduce the amount of Ge-68 ions.

[0116] The kit is a radiolabeled imaging product that can be injected directly into the patient. 68 Ga-N To obtain the eoB injection solution, 68 Ge / 68 H provided by Ga generator Cl 68 It must be used in combination with a Ga solution.

[0117] Equivalent to the amount of radioactivity administered, 68 The volume of Ga-NeoB injection solution is determined by the generator The current radioactivity provided by the Based on this, calculate the estimated time of injection accordingly. 68 Ga-NeoB injection solution is a single-dose It is a thing.

[0118] Vial 1 contains 50 μg of active ingredient packed in a 10 mL glass vial. It is a powder for injection solution containing NeoB.

[0119] The composition of Vial 1 is provided in Table 2.

[0120] [Table 2]

[0121] The composition of Vial 2 is provided in Table 3.

[0122] [Table 3]

[0123] 2. Drug development As described above, Vial 1 (NeoB, 50 μg, powder for injection solution) Radiopharmaceutical kit also containing reaction buffer (vial 2) and accessory cartridges It's a part of it.

[0124] The kit is a radiolabeled imaging product that can be injected directly into the patient. 68 Ga-N To obtain the eoB injection solution, 68 Ge / 68 H provided by Ga generator Cl 68 It must be used in combination with a Ga solution.

[0125] 2.1 Ingredients of the formulation The formulation contains NeoB as the active ingredient, and gentisic acid, mannitol, and Kolliphor HS 15.

[0126] 2.1.1 Drug Substance The active ingredient is NeoB peptide, PABZA, as shown in formula (I) below. -7 amino acid long amino acid covalently linked to a chelator (DOTA) by a DIG linker It is an acid sequence. [ka]

[0127] 2.1.2 Additives The excipients selected for the composition of vial 1 are intended to ensure the stability of the active ingredient in the final drug product. To maintain the stability of the formulation, to ensure safety and efficacy of the formulation, and further to During the procedure,68 To obtain the required radiochemical purity of the Ga-NeoB solution, additional The selected additives result in a formulation with the required technical characteristics of the drug.

[0128] Gentisic acid, a non-pharmacopoeial excipient with specific functions, is added to the formulation composition and, after dissolution, It correlates with the purity and stability of the resulting radiolabeled imaging product.

[0129] A brief description of each additive is provided below.

[0130] Mannitol Mannitol is used as a bulking agent. Peptide drugs are very potent and therefore require very In the absence of bulking agents, processing of the product is technically The bulking agent is not suitable from the viewpoint of processing the pharmaceutical product and producing a suitable lyophilized product. This makes it possible.

[0131] Gentisic acid Gentisic acid is a pharmacopoeial agent used as an antioxidant in pharmaceutical compounding. It is an additive that does not contain

[0132] Kolliphor HS 15 (Macrogol 15-hydroxystearic acid) Kolliphor HS 15 is a water-soluble nonionic surfactant used in parenteral pharmaceutical preparations. As a solubilizer, Kolliphor HS 15 is particularly suitable for parenteral and and oral dosage forms.

[0133] Nonspecific binding of peptides used as active ingredients in NeoB radiopharmaceutical kits For this reason, Kolliphor HS 15 adheres to glass and plastic surfaces. It is used as a surfactant for peptides that have a tendency to break down. 6 8 There is no risk of interference during labeling with Ga.

[0134] 2.2 Formulation 2.2.1 Drug Development Drug development can be performed without any treatment of the eluate or any additional purification steps. , commercially available 68 Ge / 68 Direct elution from Ga generator Composition of the reaction mixture that can enable simple dissolution-based labeling of DOTA-peptides This was carried out with the aim of identifying

[0135] The goal is to develop a bone marrow biosynthetic target for use as a radiotracer for the detection of GRPR-positive tumors. The aim of this study was to develop a vesin-like peptide antagonist (NeoB).

[0136] Vial 1 during the radiolabeling procedure 68 Peptides are radiolabeled with Ga as active ingredients. It is a freeze-dried powder containing methicillin-containing benzodiazepine.

[0137] The first attempt to develop a suitable drug for NeoB (vial 1) was to prepare it on a laboratory scale. This involved testing on the bulk solution prior to the sterilization and freeze-drying process.

[0138] The development work has targeted radiochemical purity as follows: 68 Ga radiolabeled Ne To obtain the final product leading to the oB product, the selection of the appropriate additives is important with respect to the characteristics of the peptide. I saw it. · 68 Ga-NeoB (HPLC) →>92% Free 68 Ga 3+ (HPLC) →≦2% ·Non-complex formation 68 Ga 3+ Seed (ITLC)→≦3%.

[0139] The components selected for the final drug are as follows:

[0140] [Table 4]

[0141] The development work, including the relevant studies carried out, is described below, beginning with the amount of active ingredient and and the selection of suitable additives.

[0142] 2.2.1.1 Selection of peptide amount 1850MBq 68 Ge / 68 Eluate produced by Ga generator and formic acid buffer Using the PHARMA® 1000, increasing amounts of NeoB peptide (15 μg to 100 μg) were injected. and greater than 98% by HPLC and 97% by ITLC. 68 Incorporation of Ga The labeling procedure was tested to identify the minimum amount of peptide required to achieve the desired results. Based on the results summarized in [1], the most reproducible results with good radiochemical purity are The lower amount of peptide is 25 μg.

[0143] [Table 5]

[0144] In parallel, various peptide doses were also tested in in vivo biodistribution experiments. Briefly, using a mouse prostate cancer model, we identified two peptides with different masses. The doses were compared: 10 pmol vs. 200 pmol; the amount of total radioactivity injected was The injection of radiolabeled NeoB resulted in a higher penetration rate. When a high peptide mass dose was used (200 pmol), it resulted in increased accumulation in tumors. At the same time, uptake in non-target organs (such as the pancreas) was significantly higher at higher peptide mass doses. (200 pmol), which was quite low. Therefore, from these preclinical evaluations, The peptide mass dose is associated with reduced uptake in non-target organs (particularly in this case, the pancreas). It has been demonstrated that this is preferable because it is related to

[0145] Radiolabeling studies performed (described in Table 5) and doses of higher peptide masses In vivo biodistribution shown to ensure better efficacy and safety profile of the compound Based on the experiment, the final amount of peptide selected for inclusion in vial 1 was 50 μg. did.

[0146] The drug development work also focused on selecting surfactants, antioxidants, and bulking agents. The radiolabeling procedure was also carefully evaluated.

[0147] 2.2.1.2 Selection of important additives ·Surfactant selection Run to confirm the drug content of vial 1 (NeoB 50 μg, powder for injection solution). During the tests, the peptides showed a particular tendency to stick to glass and plastic surfaces. This phenomenon is called nonspecific binding (NSB). Peptides bind more readily than small molecules. Uncharged peptides, in particular, often exhibit greater NSB problems than non-charged peptides, which are more resistant to plastics. The causes may be various: physical / chemical properties, foundation Lewaels interactions, ionic interactions, therefore, including surfactants and solubilizers, The addition of additives known to reduce NSB was evaluated.

[0148] Organic solvents may enhance solubility and prevent adsorption. Ethanol, for example, To enhance the solubility of highly lipophilic tracers, or vials, membrane fillers For use in radiopharmaceutical injections to reduce adsorption to syringes and syringes. Ethanol is not compatible with the freeze-drying process, so it is recommended to use Neo In the case of powder B, this is not an option.

[0149] Human serum albumin (HSA) is also often used as a stabilizer to prevent surface adsorption. However, this additive is not suitable due to its thermal instability. not present.

[0150] Another possible approach to reduce peptide nonspecific binding is to use detergents (e.g., Polysorbate 20, Polysorbate 80, Pluronic F-68, Triolein The ionic surfactants used were sorbitan phosphate. 68 May interfere with Ga labeling Since nonionic surfactants are difficult to obtain, research into nonionic surfactants has received particular attention.

[0151] Kolliphor HS 15, Kolliphor K188, Tween 20 Non-ionic surfactants such as Tween 80 and polyvinylpyrrolidone K10 are effective against oral and is commercially available as a solubilizing additive in injectable drugs.

[0152] The most common ingredients that can be used in the composition of NeoB powder (Vial 1) for injection solution To assess the suitability of suitable agents, peptide adhesion tests are performed with various surfactants. (See results in Table 6 below).

[0153] Hydroxypropyl beta-cyclodextrin, alone or in combination with a surfactant, may also be used. As reported below, hydroxypropyl β-cyclodextrin The presence of phosphorus had only a limited positive effect on peptide attachment. As demonstrated in subsequent studies (see also Section 2.2.1.3 Radiolabeling Procedures) The presence of surfactant and hydroxypropyl β-cyclodextrin was does not improve the radiochemical purity of the final product compared to a drug containing Additionally, hydroxypropyl beta-cyclodextrin was not included in the final formulation.

[0154] [Table 6]

[0155] The best results for peptide attachment were obtained with Kolliphor HS 15 and Tw The two additives were used to determine the final amounts included in the kit. The results obtained were satisfactory in terms of radiochemical purity and peptide attachment. It was something like that.

[0156] [Table 7]

[0157] Polysorbate (Tween 20) is highly soluble in water and is easily absorbed by the presence of oxygen, metal ions, peroxides, or heat. Autoxidation caused by increased temperature, cleavage at the ethylene oxide subunit, and may undergo hydrolysis of the fatty acid ester bond, ultimately resulting in Kolliph or HS 15 was chosen.

[0158] The lowest peptide deposition was observed when using 0.5 mg of Kolliphor HS 15. This is obtained by adding the selected Kolliphor HS in the final composition of the formulation. Makes 15 servings.

[0159] Antioxidant selection The presence of radical scavengers protects NeoB from radiolysis due to its antioxidant properties. Allows you to protect.

[0160] For development studies, the inventors have identified antioxidants for use in radiopharmaceutical preparations. Gentisic acid and ascorbic acid were tested as potential inhibitors. To identify the lowest amount of antioxidant that can exert the desired protective function without I went to see it.

[0161] Radiolabeling is primarily performed using the most suitable antioxidant and DOTA-peptide. 68 Incorporation of Ga The amount of antioxidant was varied and other parameters were examined to identify a concentration that would not interfere with the absorption. As shown in the table below, gentisic acid 68 Ga It is the best antioxidant as it does not interfere with the incorporation of The amount of gentisic acid selected is 200 μg.

[0162] [Table 8]

[0163] [Table 9]

[0164] ·Selection of bulking agent The drug is finally completed by the addition of bulking agents required in the process of freeze-drying the product. It was accomplished.

[0165] Among the bulking agents commonly proposed for lyophilization of peptides, formulation manufacturers have Cholesterol and mannitol were tested.

[0166] [Table 10]

[0167] Mannitol is the most commonly used lyophilisate and is also used in freeze-drying processes. To produce a cake with good characteristics in terms of appearance, stability, and moisture in the process Mannitol was chosen because it is known to have the following properties: and has been described as a good scavenger of OH radicals.

[0168] 2.2.1.3 Radiolabeling procedure Based on a two-vial design, a three-step labeling procedure was developed as follows: 1. Heat block (make sure the temperature has already reached 95°C before starting the elution) In this regard, 68 Ge / 68 in HCl provided by a Ga generator 68 The lyophilized drug (vial 1) is directly dissolved in the Ga solution. 2. Add the required volume of reaction buffer (vial 2). 3. Heat at 95°C for at least 7 minutes (do not exceed 10 minutes).

[0169] at this point 68 The Ga-NeoB solution is ready for administration.

[0170] During the development of the labeling procedure, various time and temperature conditions were tested.

[0171] The dependence of labeling efficiency on temperature is 68 A good combination was achieved in a time frame compatible with the short half-life of Ga. This study aimed to identify values that indicate congestion.

[0172] 68 Incorporation of Ga into the DOTA chelating moiety requires heating to be complete. It is known that...

[0173] The first tested labeling conditions were as follows: various reaction times (3, 5, and 7 min) ) at 80, 85, and 95°C. These tests were carried out using the following chemicals: Peptide (50 μg), Mannitol (20 mg), Gentisic acid (0.2 mg), Kolliphor HS 15 (0.5 mg), · Hydroxypropyl beta-cyclodextrin (3 mg).

[0174] The drugs tested in these initial studies were solubilizers (hydroxypropyl β-cyclohexyl benzoate, hydroxypropyl ... However, later during development, similar tests were conducted on When performed with the same agent but without hydroxypropyl β-cyclodextrin, good release was observed. In addition, the attachment of peptides was also performed by hydroxypropylating the peptides. It was shown that the absence of propyl beta-cyclodextrin did not affect the Therefore, hydroxypropyl beta-cyclodextrin was not included in the final drug product. At 85°C and 85°C, radiometric analysis showed sufficient incorporation in 7 minutes.

[0175] At 95°C, incorporation is only complete after 7 minutes.

[0176] Based on these observations, 7 minutes at 95°C is the most conservative labeling condition, with a ±15 Even when the temperature fluctuates over a 100°C range, over 98% of the recombination is achieved without significant fragmentation. It was shown that the loading can be guaranteed.

[0177] [Table 11]

[0178] Moreover, to increase the robustness of the labeling procedure, the reaction buffer ( Vial 2) was added (only after adding the reaction buffer did the labeling reaction begin). (The procedure was carried out at 95°C.) The results shown in Table 11 demonstrate good radiochemical purity. It is confirmed that the same results can be obtained under these conditions.

[0179] [Table 12]

[0180] 2.2.1.4 Final selected drug (Vial 1) Based on all the development studies mentioned above, 50 μg NeoB, powder for injection solution ( The final composition of vial 1) is as follows:

[0181] [Table 13]

[0182] The final drug product will be tested for radiolabeled product to confirm the results obtained during development. I tried it.

[0183] [Table 14]

[0184] As shown in Table 13, good results were obtained by both ITLC and HPLC (>92%). The radiochemical purity results were based on three independent radiolabeling studies performed on the final drug. It is important to note that the free gallium (by HPLC) was always below 2%. Finally, peptide attachment to glass is also important for these radiolabeled reagents. During the experiment, the presence of Kolliphor HS15 was tested to determine whether peptide attachment was permissive. It was determined that this was necessary to maintain the bell.

[0185] 2.2.1.5 Quality Standards Evaluation To precisely define quality criteria, a series of preliminary experiments was performed as summarized below. .

[0186] labeled pH The labeled pH is 68 Regarding the radiolabeling efficiency of DOTA-peptide with GaCl3, Due to its specific chemical behavior, it is one of the important parameters for obtaining good results. To determine the pH range in which knowledge gives good results, 68 Gallium-labeled Ne The oB drugs were tested while maintaining a pH range between 3.0 and 4.0. The volume of reaction buffer added was varied and tested while other parameters were kept constant. As shown in Tables 14 and 15, the pH variation within the range 3.0 to 4.0 The radiolabeled product obtained meets the standards for radiochemical purity.

[0187] [Table 15]

[0188] [Table 16]

[0189] Gentisic acid versus volume radioactivity The exam is 68 Ge / 68 The highest Ga generator can provide at that time of volumetric radioactivity 68 When labeling is performed with GaCl3, radiolytic scavengers This study was carried out to evaluate the effect of gentisic acid as the highest possible volumetric radiation Fractional elution was performed to ensure the highest radioactivity; only the fraction with the highest radioactivity was used for labeling. did.

[0190] The protective effect was observed in the presence of various amounts of gentisic acid (0.20 mg and 0.35 mg). The results (Table 1) were confirmed by monitoring peptide fragmentation over a period of time. 6) found almost the same positive effects in both studies. Therefore, most of the gentisic acid is sufficient to achieve a good level of protection from radiolysis. A low dose (200 μg) was chosen.

[0191] [Table 17]

[0192] In addition, lower amounts of gentisic acid may still be used as an antioxidant in the final drug. To test whether it could work, the first test was to administer 0.1 mg of gentamicin. The results of radiolabeling experiments performed under these conditions are shown in Table 17. However, good radiochemical purity is obtained even in the presence of lower amounts of gentisic acid. Nevertheless, it is confirmed that good radiochemical purity can be obtained from the generator. To ensure that higher radioactivity of the genotype is obtained, The amount of thidic acid was conservatively kept at 200 μg.

[0193] [Table 18]

[0194] Scale-up batch 68 Test results of Ga radiolabeled products Table 18 shows the results of two radiolabeled experiments performed with scaled-up batch NeoB vial 1. The results are summarized below. 68 Ga-NeoB meets radiochemical purity standards up to 4 hours after the end of the radiolabeling reaction This shows that.

[0195] [Table 19]

[0196] References 1. Sah BR, Burger IA, Schibli R, Friebe M, Di nkelborg L, Graham K, Borkowski S, Bacher-S Tier C, Valencia R, Srinivasan A et al:Dos imetry and First Clinical Evaluation of the New 18F-Radiolabeled Bombesin Analog ue BAY 864367 in Patients with Prostate Cancer.J Nucl Med 2015,56(3):372-378. 2.Kahkonen E,Jambor I,Kemppainen J,Lehti o K,Gronroos TJ,Kuisma A,Luoto P,Sipila HJ,Tolvanen T,Alanen K et al:In vivo ima ging of prostate cancer using[68Ga]-labe led bombesin analog BAY86-7548.Clin Canc er Res 2013,19(19):5434-5443. 3.Maina T,Bergsma H,Kulkarni HR,Mueller D,Charalambidis D,Krenning EP,Nock BA,de Jong M,Baum RP:Preclinical and first cl inical experience with the gastrin-relea sing peptide receptor-antagonist[(68)Ga] SB3 and PET / CT.Eur J Nucl Med Mol Imagin g 2016,43(5):964-973. 4.Dimitrakopoulou-Strauss A,Hohenberger P,Haberkorn U,Macke HR,Eisenhut M,Straus s LG:68Ga-labeled bombesin studies in pa tients with gastrointestinal stromal tum ors:comparison with 18F-FDG.J Nucl Med 2 007,48(8):1245-1250. 5.Velikyan I,Xu H,Nair M,Hall H:Robust l abeling and comparative preclinical char acterization of DOTA-TOC and DOTA-TATE.N ucl Med Biol 2012,39(5):628-639.

Claims

1. Radioisotopes, preferably ６８ Ga, ６７ Ga, or ６４ Cu induces gastrin release 1. A method for labeling a peptide receptor (GRPR) antagonist, comprising: i. providing a first vial containing said GRPR antagonist in dry form; P ii. adding the radioisotope solution into the first vial, thereby obtaining a solution of said GRPR antagonist with a radioisotope; iii. Mixing the solution obtained in ii with at least one buffer and and for a period of time sufficient to obtain the radioisotope-labeled GRPR antagonist. incubating, and iv. Optionally, adjusting the pH of the solution. A method comprising:

2. The first vial in step i. preferably contains both of said GRPR aerosols in dry form.

10. The method of claim 1, wherein the reaction vial comprises an antagonist and a buffer.

3. Step iii is to subject the solution obtained in step ii to at least one reaction mixture containing a buffer. and mixing the radioisotope-labeled GRPR antagonist with the solution.

2. The method of claim 1, comprising incubating it for a period of time sufficient to obtain

4. The GRPR antagonist has the formula (I): 【Chemical 1】 (DOTA-(p-aminobenzylamine-diglycolic acid))-[D-Phe-Gln -Trp-Ala-Val-Gly-His-NH-CH[CH ２ -CH(CH ３ ) ２ ] ２ The method according to any one of claims 1 to 3, wherein the compound is

5. The GRPR antagonist is administered in an amount between 20 and 60 μg, typically 50 μg, The method of any one of claims 1 to 4, contained in a first vial.

6. The first vial preferably contains between 50 and 250 μg, typically 200 μg.

6. The composition according to claim 1, further comprising gentisic acid as a radiolysis protectant. The method described.

7. The first vial contains, for example, between 10 and 30 mg, typically 20 mg, and a bulking agent The method according to any one of claims 1 to 6, further comprising mannitol as an antioxidant.

8. the first vial contains, for example, between 250 and 750 μg, typically 500 μg; Claims 1 to 7 further comprising macrogol 15 hydroxystearic acid as a surfactant.

10. The method according to any one of the preceding claims.

9. Injectable solution for in vivo detection of tumors by diagnostic imaging in a subject in need thereof; 10. The method of claim 1 for use as a pharmaceutical composition for the treatment of ulcerative colitis. A solution containing a radiolabeled GRPR antagonist, obtained or obtained.

10. Injectable solution for in vivo detection of tumors by diagnostic imaging in a subject in need thereof; or obtainable by any one of the methods of claims 4 to 8 for use as a Obtained, ６８ A solution comprising a compound of formula (I) according to claim 4 which is labeled with Ga.

11. The following components in dry form: i. The following formula: C-S-P (where, C is a chelating agent capable of chelating said radioisotope; S is an optional spacer covalently bonded between C and the N-terminus of P; P preferably has the general formula: Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Z (Xaa1 is absent or is an amino acid residue Asn, Thr, Phe, 3-(2-thienoyl) 4-chlorophenylalanine (Cpa), α-naphthylalanine (α-phenylalanine), α-Naphthylalanine (α-Nal), β-naphthylalanine (β-Nal), 1,2,3,4-tetrahydrofuran Dolonorharman-3-carboxylic acid (Tpi), Tyr, 3-iodo-tyrosine (o-I -Tyr), Trp, and pentafluorophenylalanine (5-F-Phe) (all L- or D-isomers); Xaa2 is Gln, Asn, or His; Xaa3 is Trp or 1,2,3,4-tetrahydronorharman-3-carboxylic acid ( Tpi); Xaa4 is Ala, Ser, or Val; Xaa5 is Val, Ser, or Thr; Xaa6 is Gly, sarcosine (Sar), D-Ala, or β-Ala; Xaa7 is His or (3-methyl)histidine (3-Me)His; Z is —NHOH, —NHNH2, —NH-alkyl, —N(alkyl)2, and —O— alkyl or Z is selected from 【Chemistry 2】 where X is NH (amide) or O (ester), and R1 and R2 may be the same or different and may be selected from the group consisting of a proton, an optionally substituted alkyl group, alkyl, optionally substituted alkyl ether, aryl, aryl ether, or aryl alkyl, halogen, hydroxyl, hydroxyalkyl, amine, amino, amide do), or amide substituted aryl or heteroaryl groups a GRPR antagonist of the formula (I) which is a GRPR peptide antagonist of the formula (I); ii. Radiolysis protectants, such as gentisic acid; iii. Bulking agents, such as mannitol; and iv. Optionally, a surfactant, such as macrogol 15 hydroxystearic acid 1. A powder for injection solution, comprising:

12. The GRPR antagonist has the formula (I): 【Chemistry 3】 12. Powder for injection solutions according to claim 11, which is a compound of formula:

13. Consists of: - an amount between 20 and 60 μg, typically 50 μg, of the following formula (I); 【Chemistry 4】 - gentisic acid in amounts of 50 and 250 μg, typically 200 μg; - an amount between 10 and 30 mg, for example 20 mg of mannitol, - an amount between 250 and 750 μg, for example 500 μg of macrogol 15 hydroxystearate phosphoric acid 13. A powder for injection solutions according to any one of claims 11 to 12, comprising:

14. 1. A kit comprising: i. The following components in dry form: a compound of formula (I): 【Chemistry 5】 - radiolysis protectants, e.g. gentisic acid, optionally a bulking agent, such as mannitol, and - optionally a surfactant, such as macrogol 15 hydroxystearic acid a first vial having: ii. a second vial containing at least one buffer, preferably in dry form; and to iii. Optionally, dissolving the radioisotopes generated by the radioisotope generator. Included cartridge for discharging A kit for carrying out the method of claim 4, comprising:

15. A kit comprising: i. The following components in dry form: a compound of formula (I): 【Chemistry 6】 - radiolysis protectants, e.g. gentisic acid, - optionally a bulking agent, e.g. mannitol, - optionally a surfactant, such as macrogol 15 hydroxystearic acid; and at least one buffer, preferably in dry form a single vial having ii. Optionally, eluting radioisotopes generated by a radioisotope generator. Included cartridge for A kit for carrying out the method of claim 4, comprising:

16. The first or single vial contains the following components: - an amount between 20 and 60 μg, typically 50 μg, of the following compounds of formula (I); 【Chemistry 7】 - gentisic acid in amounts of 50 and 250 μg, typically 200 μg; - mannitol in an amount between 10 and 30 mg, for example 20 mg, and - optionally in an amount between 250 and 750 μg, for example 500 μg of macrogol 15 hydroxybenzoates Roxystearic Acid The kit according to any one of claims 14 to 15, comprising:

17. All components of the first, second, or single vial are in dry form. The kit according to any one of claims 14 to 16.

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