Methods for radiolabeling GRPR antagonists and kits therefor

Abstract

The present invention relates to methods and kits for radiolabeling GRPR antagonists, such as NeoB. In particular, the present invention relates to methods for radiolabeling GRPR antagonists, such as NeoB, with radioisotopes, preferably 68 Ga, 67 Ga, or 64 The present invention relates to a method for labeling a gastrin-releasing peptide receptor (GRPR) antagonist with Cu, comprising the steps of: i. providing a first vial containing the GRPR antagonist in a dried form; ii. adding a solution of the radioisotope into the first vial, thereby obtaining a solution of the GRPR antagonist carrying the radioisotope; iii. mixing the solution obtained in ii. with at least one buffer and incubating it for a period of time sufficient to obtain the GRPR antagonist labeled with the radioisotope; and iv. optionally, adjusting the pH of the solution.

Description

[Technical field]

[0001] The present disclosure relates to methods and kits thereof for radiolabeling GRPR antagonists, such as NeoB. [Background technology]

[0002] Bombesin was first isolated from the European frog, Bombina bombina, and demonstrated to be similar to mammalian gastrin-releasing peptide (GRP) and neuromedin B (NMB): Erspamer, V. Discovery, Isolation, and Characterization of Bombesin-like Peptides. Ann NY Acad Sci 547:3-9, 1988; Jensen, RT; Battey, JF; Spindel, ER; Benya, RV International union of pharmacology. LXVIII. Mammalian bombesin 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, regulates numerous functions of the gastrointestinal and central nervous systems, including gastrointestinal hormone release, smooth muscle cell contraction, and epithelial cell proliferation. Gastrin releasing peptide is a potent mitogen for physiological and neoplastic tissues, and gastrin releasing peptide may be involved in growth dysregulation and carcinogenesis.

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

[0005] GRPR is highly overexpressed in prostate cancer, and studies in human prostate cancer cell lines and xenograft models have demonstrated both high affinity (nM levels) and high tumor accumulation (%ID / g), but the relative expression of GRPR across early to late stage aggressive disease has not yet been fully characterized [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 were determined by immunohistochemistry in randomly selected colon cancer samples, including LN and metastatic lesions. More than 80% of the samples aberrantly expressed GRP or GRPR, and more than 60% expressed both GRP and GRPR, whereas no expression was observed in the adjacent normal healthy epithelium [Scopinaro F, et al. Cancer Biother Radiopharm 2002,17(3):327-335].

[0007] GRP is normally present in pulmonary neuroendocrine cells and plays a role in stimulating lung development and maturation. However, it also appears to be involved in growth dysregulation and carcinogenesis. Stimulation of GRP leads to increased release of epidermal growth factor receptor (EGFR) ligands, which subsequently activates EGFR and mitogen-activated protein kinase downstream pathways. Using non-small cell lung cancer (NSCLC) cell lines, it was confirmed that both EGF and GRP stimulated NSCLC proliferation, and inhibition of EGFR or GRPR led to cell death [Shariati F, et al. Nucl Med Commun 2014,35(6):620-625].

[0008] In nuclear medicine, peptide receptor agonists have long been the ligands of choice for the development and use of tracers. The rationale behind the use of agonist-based constructs was the internalization of the receptor-radioligand complex, allowing high accumulation of radioactivity in the target cells. In the case of radiometal-labeled peptides, efficient receptor-mediated endocytosis in response to agonist stimulation leads to high in vivo radioactivity accumulation in the targeted tissue, a key prerequisite for optimal imaging of malignant diseases. However, a paradigm shift occurred when receptor-selective peptide antagonists showed favorable biodistribution, including significantly higher in vivo tumor accumulation, compared to highly potent agonists. Further advantages of GRPR antagonists are that they are safer to use in clinical practice since no acute biological side effects are expected from their use and that the tracer dose is not as high considering the current diagnostic and potentially higher doses for potential therapeutic purposes [Stoykow C, et al. Theragnostics 2016,6(10):1641-1650].

[0009] In preclinical models, 68 Ga]-NeoB and [ 177 Lu]-NeoB([ 68 Ga]-NeoBOMB1 and [177 Lu]-NeoBOMB1) showed high affinity for GRPR expressed in breast, prostate, and gastrointestinal stromal tumors (GIST), and low internalization upon binding to the specific receptor. The ability of the radiolabeled peptide to target GRPR-expressing tumors was confirmed in in vivo imaging and biodistribution studies in animal models [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 An optimized method for labeling NeoB with Cu and thereby obtaining a labeled NeoB solution for imaging diagnostic purposes of GRPR-positive tumors in human patients has not been developed. In particular, a method for labeling NeoB with Cu for intravenous injection in human subjects in need thereof has not been developed. 68 There is a need for a rapid, efficient, and safe procedure that would provide labeled GRPR antagonists of high radiochemical purity, such as [Ga]NeoB. Summary of the Invention [Means for solving the problem]

[0011] A first aspect of the present disclosure relates to a method for the preparation of radioisotopes, preferably 68 Ga, 67 Ga, or 64 1. A method for labeling a Gastrin Releasing Peptide Receptor (GRPR) antagonist with Cu, comprising: i. providing a first vial containing said GRPR antagonist in a dried form; ii. adding a solution of said radioisotope into said first vial, thereby obtaining a solution of said GRPR antagonist carrying said radioisotope; iii. mixing the solution obtained in ii. with at least one buffer and incubating it for a period of time sufficient to obtain said radiolabeled GRPR antagonist; and iv. Optionally, adjusting the pH of the solution. The present invention relates to a method comprising the steps of:

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

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

[0014] In other specific embodiments, the radioisotope is 64 Cu and has a radiochemical purity of at least 90% as measured by HPLC, and optionally is free 64 The percentage of Cu2+ (by HPLC) is less than 2% and / or uncomplexed 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 relates to a solution comprising a radioisotope-labeled GRPR antagonist obtainable or obtainable by the method disclosed herein for use as an injection for in vivo detection of tumors by diagnostic imaging in a subject in need thereof.

[0017] Consists of the following components in dry form: i. The 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 selected from the group consisting of the amino acid residues Asn, Thr, Phe, 3-(2-thienyl)alanine (Thi), 4-chlorophenylalanine (Cpa), α-naphthylalanine (α-Nal), β-naphthylalanine (β-Nal), 1,2,3,4-tetrahydronorharman-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 selected from -NHOH, -NHNH, -NH-alkyl, -N(alkyl) and -O-alkyl; or Z is [ka] wherein X is NH (amide) or O (ester), and R1 and R2 are the same or different and are selected from a proton, an optionally substituted alkyl, an optionally substituted alkyl ether, an aryl, an aryl ether, or an alkyl, halogen, hydroxyl, hydroxyalkyl, amine, amino, amido, or amide substituted aryl or heteroaryl group; 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 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 of 50 and 250 μg, typically 200 μg; and iii. Mannitol in an amount between 10 and 30 mg, for example 20 mg; and iv. An amount between 250-750 μg, e.g. 500 μg of macrogol 15 hydroxystearic 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. a first vial, optionally having a surfactant, such as macrogol 15 hydroxystearic acid; and ii. a second vial containing at least one buffering agent, preferably in dry form; and iii. Optionally, an accessory cartridge for eluting the radioisotope generated by the radioisotope generator. 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 buffering agent, preferably in dry form; and ii. Optionally, an accessory cartridge for eluting the radioisotope generated by the radioisotope generator. The vial includes 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 an amount of 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 The first or single vial may include DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0022] Generally, the present disclosure relates to radioisotopes, preferably 68 Ga, 67 Ga, or 64 1. A method for labeling a Gastrin Releasing Peptide Receptor (GRPR) antagonist with Cu, comprising: (i) providing a first vial containing said GRPR antagonist in a dried form; (ii) adding a solution of said radioisotope into said first vial, thereby obtaining a solution of said GRPR antagonist carrying said radioisotope; (iii) mixing the solution obtained in ii. with at least one buffer and incubating it for a period of time sufficient to obtain the radioisotope-labeled GRPR antagonist; (iv) optionally adjusting the pH of the solution The present invention relates to a method comprising the steps of:

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

[0024] Preferred radiolabeled GRPR antagonists obtained by the disclosed methods are radioisotopes suitable for use as contrast agents for PET / CT, SPECT, or PET / MRI imaging, preferably 68 Ga, 67 Ga, or 64 In a preferred embodiment, the compound is a NeoB compound labeled with Cu. 67 Ga is used for SPECT imaging, and 68 Ga and 64 Cu is used for PET imaging such as PET / CT or PET / MRI.

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

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

[0027] definition The phrases "treatment" and "treating" include amelioration or cessation of a disease, disorder, or a symptom thereof. In particular, with respect to the treatment of tumors, the term "treatment" may refer to inhibition of tumor growth or reduction in tumor size.

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

[0029] As used herein, "PET" stands for Positron Emission Tomography.

[0030] As used herein, "SPECT" stands for single photon emission computed tomography.

[0031] As used herein, "MRI" stands for Magnetic Resonance Imaging.

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

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

[0034] As used herein, the terms "substituted" or "optionally substituted" refer to a group that is optionally substituted with one or more substituents selected from the following, in a number ranging from zero to the total number of open valences on the aromatic ring structure: halogen, -OR', -NR'R'', -SR', -SiR'R''R''', -OC(O)R', -C(O)R', -COR', -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'''' may be independently selected from hydrogen, alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl. When a compound of the present disclosure includes more than one R group, for example, each R group is independently selected, as are each of these groups when more than one R', R'', R''', and R'''' groups are present.

[0035] As used herein, the term "alkyl," alone or as part of another substituent, refers to a straight or branched chain alkyl functional group having 1 to 12 carbon atoms. Suitable alkyl groups include 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), and hexyl and its isomers (e.g., n-hexyl, iso-hexyl).

[0036] As used herein, the term "heteroaryl" refers to a polyunsaturated aromatic ring system containing 5-10 atoms, having a single ring or multiple aromatic rings fused or covalently bonded to one another, in which at least one ring is aromatic and at least one ring atom is a heteroatom selected from N, O, and S. The nitrogen and sulfur heteroatoms may optionally be oxidized, and the nitrogen heteroatom may optionally be quaternized. Such rings may be fused to aryl, cycloalkyl, or heterocyclyl rings. Non-limiting examples of such heteroaryls include: furanyl, thiophenyl, pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, oxatriazolyl, thiatriazolyl, pyridinyl, pyrimidyl, pyrazinyl, pyridazinyl, oxazinyl, dioxinyl, thiazinyl, triazinyl, indolyl, isoindolyl, benzofuranyl, isobenzofuranyl, benzothiophenyl, isobenzothiophenyl, indazolyl, benzimidazolyl, benzoxazolyl, purinyl, benzothiadiazolyl, quinolinyl, isoquinolinyl, cinnolinyl, quinazolinyl, and quinoxalinyl.

[0037] As used herein, the term "aryl" refers to a polyunsaturated aromatic hydrocarbyl group having a single ring or multiple aromatic rings fused together, containing 6 to 10 ring atoms, where at least one ring is aromatic. The aromatic ring may optionally contain 1 to 2 additional rings (cycloalkyl, heterocyclyl, or heteroaryl as defined herein) fused thereto. Suitable aryl groups include phenyl, naphthyl, and phenyl rings fused to heterocyclyls such as benzopyranyl, benzodioxolyl, benzodioxanyl, and the like.

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

[0039] As used herein, the term "optionally substituted fatty chain" refers to an optionally substituted fatty chain having 4 to 36 carbon atoms, preferably 12 to 24 carbon atoms.

[0040] As used herein, the term "chelating agent" refers to a molecule that has functional groups, such as amine or carboxyl groups, suitable for complexing with a radioisotope via a non-covalent bond.

[0041] As used herein, the term "radiolysis protector" refers to a stabilizer that protects organic molecules from radiolysis, for example when gamma rays emitted from a radionuclide break the bonds between the atoms of the organic molecules and radicals are formed, which are then scavenged by the stabilizer, which prevents the radicals from undergoing any other chemical reactions that may result in undesirable, possibly useless, or even toxic molecules. Therefore, these stabilizers are also called "free radical scavengers" or "radical scavengers" for short. Other alternative terms for these stabilizers are "radiostability enhancers", "radiolysis stabilizers", or simply "quenchers".

[0042] As used herein, the term "radiochemical purity" refers to that percentage of a given radionuclide that is present in a given chemical or biological form. Radiochromatographic methods, such as HPLC or instant thin layer chromatography (iTLC), are the most commonly accepted methods for determining radiochemical purity in nuclear pharmacy.

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

[0044] (i) providing a first vial containing said GRPR antagonist in a dry form; 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 selected from the group consisting of the amino acid residues Asn, Thr, Phe, 3-(2-thienyl)alanine (Thi), 4-chlorophenylalanine (Cpa), α-naphthylalanine (α-Nal), β-naphthylalanine (β-Nal), 1,2,3,4-tetrahydronorharman-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 selected from -NHOH, -NHNH, -NH-alkyl, -N(alkyl) and -O-alkyl; or Z is [ka] where X is NH (amide) or O (ester), and R1 and R2 are the same or different and are selected from a proton, an optionally substituted alkyl, an optionally substituted alkyl ether, an aryl, an aryl ether, or an alkyl, halogen, hydroxyl, hydroxyalkyl, amine, amino, amido, or amide substituted aryl or heteroaryl group.

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

[0046] According to one embodiment P is DPhe-Gln-Trp-Ala-Val-Gly-His-Z; wherein Z is defined as above.

[0047] According to one embodiment P is DPhe-Gln-Trp-Ala-Val-Gly-His-Z; Z is selected from Leu-ψ(CHN)-Pro-NH and NH-CH(CH-CH(CH)), or Z is [ka] where X is NH (amide) and R is CH(CH-CH(CH) and R is (CHN)-Pro-NH, which is the same as or different from R.

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

[0049] In a particular embodiment, C is obtained by grafting a chelating agent selected from the group consisting of: [ka]

[0050] According to one embodiment, S is selected from the group consisting of: a) an aryl containing a residue of the formula: [ka] Wherein PABA is p-aminobenzoic acid, PABZA is p-aminobenzylamine, PDA is phenylenediamine, and PAMBZA is (aminomethyl)benzylamine; b) dicarboxylic acids, ω-aminocarboxylic acids, ω-diaminocarboxylic acids, or diamines of the formula: [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-amino acids, either in single or homogeneous chains of various chain lengths or in heterogeneous chains of various chain lengths, in particular [ka] GRP(1-18), GRP(14-18), GRP(13-18), BBN(1-5), 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 following formula: [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 (also referred to as NeoBOMB1) of formula (I): [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 radiolabeled NeoB2 of formula (III): [ka] (M-N4(p-aminobenzylamine-diglycolic acid)-[D-Phe-Gln-Trp-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 ProBOMB1 of formula (II): [ka] (DOTA-pABzA-DIG-D-Phe-Gln-Trp-Ala-Val-Gly-His-Leu-ψ(CH2N)-Pro-NH2).

[0055] Synthesis of Compounds of Formulae (I), (II), and (III) The compounds of formula (I), (II), and (III) can be synthesized using the methods disclosed in the reference "Positron Emission Tomography Imaging of the Gastrin-Releasing Peptide Receptor with a Novel Bombesin Analogue" ACS Omega 2019, 4, 1470-1478.

[0056] A first vial containing the GRPR antagonist. In one embodiment, the radiolabeling method employs a single vial kit, in which the first vial contains the GRPR antagonist and a buffer, both in dry form.

[0057] Alternatively, the radiolabeling method uses a two-vial kit, in this embodiment, the first vial contains the GRPR antagonist and the second vial contains a buffer.

[0058] For example, the GRPR antagonist, typically a NeoB compound, is contained in the first vial in an amount between 20-60 μg, typically 50 μg.

[0059] The first vial optionally contains additional additives such as radiolysis protectants, bulking agents, and tensioactive agents.

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

[0061] In a preferred embodiment, mannitol may be used as a bulking agent, for example in an amount between 10-30 mg, typically 20 mg.

[0062] In a preferred embodiment, macrogol 15 hydroxystearic acid may be used as a surfactant, for example in an amount between 250 and 750 μg, typically 500 μg, which advantageously reduces the non-specific adhesion of the NeoB compound to glass or plastic surfaces, thereby optimizing the rate of the labeling process.

[0063] A preferred example of the first vial (vial 1 of a two-vial kit) is provided in the Examples.

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

[0065] As used herein, a buffering agent is a buffer suitable for obtaining a pH between 3.0 and 6.0, preferably between 3.0 and 4.0, in the incubation step (iii). A "buffer for a pH between 3.0 and 6.0, preferably between 3.0 and 4.0" may advantageously be a formic acid buffer with sodium hydroxide.

[0066] The buffering agent may further be included in the first vial in embodiments using a single vial kit, or in a separate second vial in embodiments using a two-vial kit.

[0067] (ii) adding the solution of the radioisotope into the first vial; Radioisotopes for use in the radiolabeling methods include those suitable as imaging agents in PET and SPECT 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 In a preferred embodiment, 67 Ga is used for SPECT imaging, and 68 Ga and64 Cu is used for PET imaging such as PET / CT or PET / MRI.

[0069] The metal ions of such radioisotopes can form non-covalent bonds with functional groups of a chelating agent, such as a carborboxylic acid of a GRPR antagonist.

[0070] In certain embodiments, the solution of the radioisotope comprises: i. Producing a radioisotope from a parent non-radioactive element by a radioisotope generator; ii. separating the radioisotope from the parent non-radioactive element by elution in HCl as an elution solvent; 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 determines the parent element 68 From Ge 68 Producing Ga elements ii. Optionally, an element 68 Ge / 68 The generated Ga was passed through a suitable cartridge. 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 68Such methods for producing Ga are well known in the art and are described, for example, in Martinova L, et al. Gallium-68 in Medical Imaging. Curr Radiopharm. 2016; 9(3): 187-20; Dash A, Chakravarty Radionuclide generators: the prospect of availing PET radiotracers to meet current clinical needs and future research demands R Am J Nucl Med Mol Imaging. 2019 Feb 15; 9(1): 30-66.

[0073] radioactive isotope 68 The solution containing Ga may be an eluate, preferably obtained from a cyclotron production, such as described, for example, in Am J Nucl Med Mol Imaging 2014;4(4):303-310 or in BJBNelson et al. / Nuclear Medicine and Biology 80-81(2020)24-31.

[0074] Preferably, 68 Ga may be produced by a cyclotron, more preferably using a proton beam with an energy between 8 and 18 MeV, even more preferably between 11 and 14 MeV. 68 Ga uses a solid or liquid target system 68 Zn(p,n) 68 The target may be produced via the Ga reaction. 68 Zn metal or 68 After irradiation, the target is transferred to further chemical processing, 68 Ga is isolated using ion exchange chromatography. 68 Ga is dissolved in an HCl solution.

[0075] Instead, the radioisotope is67 Ga. Uses zinc (enriched or natural) or copper or germanium targets with protons, deuterons, alpha particles, or helium(III) as bombardment particles. 67 Various methods for the production of Ga are described in Helus, F., Maier-Borst, W., 1973. A comparative investigation of methods used to produce 67Ga with a cyclotron. In: Radiopharmaceuticals and Labelled Compounds, Vol. 1, IAEA, Vienna, pp. 317-324; M. L. Thakur Gallium-67 and indium-111 radiopharmaceuticals Int. J. Appl. Rad. Isot., 28 (1977), pp. 183-201; and Bjornstad, T., Holtebekk, T., 1993. Production of 67 Ga at the Oslo cyclotron. University of Oslo Report OUP8-3-1, pp. 3-5. nat Bombardment of a Ge target is also a suitable method to produce 67Ga, as described in T Horiguchi, H Kumahora, H Inoue, Y Yoshizawa Excitation functions of Ge(p,xnyp) reactions and production of 68Ge, Int. J. Appl. Radiat. Isot., 34 (1983), pp. 1531-1535.

[0076] Preferably, 67 The Ga may be produced by a cyclotron. 68 Zn(p,2n) 67 From Ga 67Such methods for producing Ga are well known in the art and are described, for example, in Alirezapour B et al. Iranian Journal of Pharmaceutical Research (2013), 12(2):355-366. More preferably, the method uses a proton beam with an energy between 10 and 40 MeV. 67 Ga uses a solid or liquid target system 67 Zn(p,n) 67 Ga or 68 Zn(p,2n) 67 The target may be produced via either the Ga reaction. 67 Zn or 68 The target was then subjected to further chemical treatment using either Zn metal or a liquid solution. 67 Ga is isolated using ion exchange chromatography and final evaporation from aqueous HCl. 67 GaCl3 is produced, which may then be added to the single vial for the labeling process.

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

[0078] Typically, 64 Cu may be produced by a cyclotron, preferably using a proton beam with energies between 11 and 18 MeV. 64 Cu can be measured using a solid or liquid target system. 64 Ni(p,n) 64 The target may be produced via a Cu reaction. 64 Ni metal or 64 After irradiation, the target was transferred to further chemical treatment, 64 Cu is isolated using ion exchange chromatography and final evaporation from aqueous HCl. 64CuCl2 is produced, which may then be added to the first vial for the labeling process.

[0079] (iii) mixing the solution obtained in step (ii) with at least one buffer and incubating it for a period of time sufficient to obtain said radiolabeled GRPR antagonist; Radiolabeling may be accomplished by irradiating a first vial containing a GRPR antagonist (e.g., a NeoB compound) with a radioisotope (typically a radioisotope as disclosed above). 68 Ga, 67 Ga, or 64 Cu) in a suitable buffer as disclosed above.

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

[0081] In a particular embodiment, the incubating step is carried out for a period comprised between 5 and 10 minutes, for example between 6 and 8 minutes, typically about 7 minutes.

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

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

[0084] In certain embodiments of the method, the method comprises the steps of: 68 The solution of Ga is i. The generator determines the parent element 68 From Ge 68 Producing Ga element; ii. Optionally, an element 68 Ga / 68 Ge was generated by passing it through a suitable cartridge. 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.

[0086] Advantageously, in certain embodiments, the simple labeling of the GRPR antagonist is commercially available without any treatment of the eluate or any additional purification steps. 68 Ge / 68In HCl produced by Ga generator 68 It may be obtained by elution of Ga.

[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 NeoB of formula (I) as defined above; 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; and iv. Between 250 and 750 μg, e.g. 500 μg of macrogol 15 hydroxystearic acid Includes.

[0089] Radiolabeling Kits of the 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 buffering agent, preferably in dry form; and iii. Optionally, an accessory cartridge for eluting the radioisotope generated by the radioisotope generator. The present invention also relates to a kit comprising:

[0090] Preferably, the 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 single vial may contain a buffer to maintain the pH between 3.0 and 4.0, for example, the second vial contains formic acid and sodium hydroxide as buffers.

[0092] Preferably, all components of the first, second or single vial are in dry form.

[0093] The radioisotope for labeling the GRPR antagonist may be provided with the kit as a ready-to-use product, i.e., for mixing and incubating with the first vial and buffer provided by the kit, or alternatively, the radioisotope may be specifically provided as: 68 Ga, 67 Ga, and 64 If it has a relatively short half-life, such as Cu, it may be mixed with the first vial and buffer and eluted from the radioisotope generator immediately prior to incubation.

[0094] Preferably, the components are in a hermetically sealed container, optionally packaged together with instructions for practicing the methods according to the present disclosure.

[0095] The kit may also be used as part of an automated system or a remotely operated mechanism that automatically performs the elution of the Gallium-69 generator and / or the subsequent mixing and heating. In this embodiment, the vial containing the GRPR antagonist (the first vial) is directly connected to the elution system and / or the heating system.

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

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

[0098] Use of kits according to the present disclosure The kit defined above may be particularly adapted for the use of the labelling methods disclosed in the previous paragraph.

[0099] Advantageously, radioisotopes (e.g. 68 Ga, 67 Ga, or 64 A solution containing a GRPR antagonist (e.g., a NeoB compound) labeled with Cu) can be obtained or is obtained by the labeling method disclosed in the previous section.

[0100] Such a solution may be ready for use as an injection for in vivo detection of tumors, for example by imaging in a subject in need thereof.

[0101] In certain embodiments, the subject is a mammal, such as, but not limited to, a rodent, a canine, a feline, or a primate. In preferred embodiments, the subject is a human.

[0102] The need for effective pharmaceutical carriers for injectable compositions is well known to those of skill in the art (see, e.g., Pharmaceutics and Pharmacy Practice, J.B. Lippincott Company, Philadelphia, PA, Banker and Chalmers, eds., pages 238-250 (1982), and ^SHP Handbook on Injectable Drugs, Trissel, 15th ed., pages 622-630 (2009)).

[0103] Typically, the solution for use as an injection provides a dose of between 150-250 MBq of [68Ga]-NeoB for administration to a subject in need thereof.

[0104] In certain embodiments, the subject in need thereof is a subject with cancer, more particularly a patient with a tumor selected from prostate cancer, breast cancer, small cell lung cancer, colon cancer, gastrointestinal stromal tumor, gastrinoma, glioma, glioblastoma, renal cell carcinoma, gastroenteropancreatic neuroendocrine tumor, esophageal squamous cell tumor, neuroblastoma, head and neck squamous cell carcinoma, and ovarian, endometrial, and pancreatic tumors that exhibit neoplasia-associated vasculature that may be GRPR positive.

[0105] Typically, PET / MRI, SPECT, or PET / CT imaging may be performed between 1 and 4 hours after administration of the radiolabeled GRPR antagonist to the subject, more preferably between 2 and 3 hours after administration of the radiolabeled GRPR antagonist to the subject.

[0106] Embodiment The following specific embodiments are disclosed: 1. Radioisotopes, preferably 68 Ga, 67 Ga, or 64 1. A method for labeling a Gastrin Releasing Peptide Receptor (GRPR) antagonist with Cu, comprising: i. providing a first vial containing said GRPR antagonist in a dried form; ii. adding a solution of said radioisotope into said first vial, thereby obtaining a solution of said GRPR antagonist carrying said radioisotope; iii. mixing the solution obtained in ii. with at least one buffer and incubating it for a period of time sufficient to obtain said radiolabeled GRPR antagonist; and iv. Optionally, adjusting the pH of the solution. The method includes: 2. The method of embodiment 1, wherein the first vial in step i. is a reaction vial containing said GRPR antagonist and a buffer, both preferably in dry form. 3. The method of embodiment 1, wherein step iii comprises mixing the solution obtained in ii. with at least one reaction solution containing a buffer and incubating it for a period of time sufficient to obtain the GRPR antagonist labeled with the radioisotope. 4. The method of any one of embodiments 1-3, wherein the solution having the radioisotope further comprises HCl. 5. The radioisotope is 68 Ga, and the radiochemical purity as measured by HPLC is at least 92%, and optionally, free 68 The percentage of Ga3+ (by HPLC) is less than 2% and / or uncomplexed 68 The method of any one of embodiments 1-4, wherein the percentage of Ga3+ species (in the ITLC) is 3% or less. 6. The radioisotope is 67 Ga, and the radiochemical purity as measured by HPLC is at least 92%, and optionally, free 67 The percentage of Ga3+ (by HPLC) is less than 2% and / or uncomplexed 67 The method of any one of embodiments 1-4, wherein the percentage of Ga3+ species (in the ITLC) is 3% or less. 7. The radioisotope is 64 Cu and has a radiochemical purity of at least 92% as measured by HPLC, and optionally free 64 The percentage of Cu2+ (by HPLC) is less than 2% and / or uncomplexed 64 The method of any one of embodiments 1-4, wherein the percentage of Cu2+ species (in the ITLC) is 3% or less. 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 selected from the group consisting of the amino acid residues Asn, Thr, Phe, 3-(2-thienyl)alanine (Thi), 4-chlorophenylalanine (Cpa), α-naphthylalanine (α-Nal), β-naphthylalanine (β-Nal), 1,2,3,4-tetrahydronorharman-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 selected from -NHOH, -NHNH, -NH-alkyl, -N(alkyl) and -O-alkyl; or Z is [ka] wherein X is NH (amide) or O (ester), and R1 and R2 are the same or different and are selected from a proton, an optionally substituted alkyl, an optionally substituted alkyl ether, an aryl, an aryl ether, or an alkyl, halogen, hydroxyl, hydroxyalkyl, amine, amino, amido, or amide substituted aryl or heteroaryl group; and 9. The method of embodiment 8, wherein P is DPhe-Gln-Trp-Ala-Val-Gly-His-NH-CH(CH2-CH(CH3)2)2. 10. The GRPR antagonist has the formula (I): [ka] DOTA-(p-aminobenzylamine-diglycolic acid)-D-Phe-Gln-Trp-Ala-Val-Gly-His-NH-CH[CH2-CH(CH3)2]2. The method of embodiment 8, wherein the NeoB compound is 11. The method of any one of embodiments 1-10, wherein the GRPR antagonist is contained in the first vial in an amount between 20 and 60 μg, typically 50 μg. 12. The method of any one of the preceding embodiments, wherein the first vial further comprises gentisic acid as a radiolysis protectant, preferably in an amount between 50 and 250 μg, typically 200 μg. 13. The method of any one of embodiments 1-12, wherein the first vial further comprises mannitol as a bulking agent, e.g., in an amount between 10 and 30 mg, typically 20 mg. 14. The method of any one of the preceding embodiments, wherein the first vial further comprises macrogol 15 hydroxystearic acid as a surfactant, for example in an amount between 250 and 750 μg, typically 500 μg. 15. The method of any one of the preceding embodiments, wherein the buffering agent is present in an amount suitable to obtain a pH of between 3.0 and 4.0 in the incubating step (iii). 16. The method of any one of the preceding embodiments, wherein the buffer comprises formic acid and sodium hydroxide. 17. The method of any one of embodiments 1-16, wherein the incubating step is carried out at a temperature between 80°C and 100°C, preferably between 90°C and 100°C, typically at about 95°C. 18. The method of any one of the preceding embodiments, wherein the incubating step is carried out for a period comprised between 5 and 10 minutes, such as between 6 and 8 minutes, typically about 7 minutes. 19. The solution of the radioisotope is i. Producing a radioisotope from a parent non-radioactive element by a radioisotope generator; ii. separating the radioisotope from the parent non-radioactive element by elution in HCl as an elution solvent; iii. collecting the eluate; thereby obtaining a solution of said radioisotope in HCl. 19. The method of any one of embodiments 1-18, wherein the eluate is obtained from 20. 68 A method for labeling a NeoB compound of formula (I) with Ga, comprising the steps of: [ka] (DOTA-(p-aminobenzylamine-diglycolic acid))-D-Phe-Gln-Trp-Ala-Val-Gly-His-NH-CH[CH2-CH(CH3)2]2 i. providing a first vial containing about 50 μg of NeoB and between 50 and 250 μg of gentisic acid in a dry form; ii. Add HCl to the first vial 68 adding a solution of Ga; iii. mixing the solution obtained in ii. with a buffer to adjust the pH to the range of 3.0 and 4.0; 68 incubating said NeoB compound for a period of time sufficient to obtain a Ga-labeled NeoB compound; iv. Optionally, adjusting the pH of the solution. The method includes: 21. The above in HCl 68 The solution of Ga is i. The generator determines the parent element 68 From Ge 68 Producing Ga element; ii. Optionally, an element 68 Ga / 68 Ge was generated by passing it through a suitable cartridge. 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 method of embodiment 20 or 21, wherein the buffer consists of 60 mg of formic acid and 56.5 mg of sodium hydroxide. 23. The method of any one of embodiments 20-22, wherein the incubating step is carried out at a temperature between 80°C and 100°C, preferably between 90°C and 100°C, typically at about 95°C. 24. The method of any one of embodiments 20-23, wherein the incubating step is carried out for a period comprised between 5 and 10 minutes, for example between 6 and 8 minutes, typically about 7 minutes. 25. A solution comprising a GRPR antagonist labeled with a radioisotope, obtainable or obtainable by the method of any one of embodiments 1 to 24, for use as an injection solution for in vivo detection of tumors by imaging diagnosis in a subject in need thereof. 26. A method obtainable or obtained by any one of the methods of embodiments 20 to 24 for use as an injection for in vivo detection of tumors by imaging in a subject in need thereof; 68 A solution containing Ga-labeled NeoB compound. 27. The solution for use according to embodiment 25 or embodiment 26, wherein the tumor is selected from tumors expressing GRPR, preferably the tumor expressing GRPR is selected from prostate cancer, breast cancer, small cell lung cancer, colon cancer, gastrointestinal stromal tumors, gastrinoma, renal cell carcinoma, gastroenteropancreatic neuroendocrine tumors, esophageal squamous cell tumors, neuroblastoma, head and neck squamous cell carcinoma, as well as ovarian tumors, endometrial tumors, and pancreatic tumors exhibiting GRPR-positive neoplasm-associated vasculature. 28. A dry form of the following components: i. The 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 selected from the group consisting of the amino acid residues Asn, Thr, Phe, 3-(2-thienyl)alanine (Thi), 4-chlorophenylalanine (Cpa), α-naphthylalanine (α-Nal), β-naphthylalanine (β-Nal), 1,2,3,4-tetrahydronorharman-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 selected from -NHOH, -NHNH, -NH-alkyl, -N(alkyl) and -O-alkyl; or Z is [ka] wherein X is NH (amide) or O (ester), and R1 and R2 are the same or different and are selected from a proton, an optionally substituted alkyl, an optionally substituted alkyl ether, an aryl, an aryl ether, or an alkyl, halogen, hydroxyl, hydroxyalkyl, amine, amino, amido, or amide substituted aryl or heteroaryl group; ii. Radiolytic protectants, such as gentisic acid; iii. bulking agents, such as mannitol; and iv. Optionally, a surfactant, such as macrogol 15 hydroxystearic acid 4. A powder for injection solution comprising: 29. A powder for injection solution according to embodiment 28, wherein said GRPR antagonist is a NeoB compound of formula (I) below: [ka] 30. A powder for injection solution according to embodiment 29, in which the NeoB compound is contained in an amount between 20 and 60 μg, typically 50 μg. 31. The powder for injection solution of any one of embodiments 28 to 30, wherein gentisic acid is contained in an amount between 50 and 250 μg, typically 200 μg. 32. The powder for injection solution of any one of embodiments 28 to 31, wherein the bulking agent is mannitol in an amount between 10 and 30 mg, for example 20 mg. 33. The powder for injection solution of any one of embodiments 28 to 32, wherein the surfactant is macrogol 15 hydroxystearic acid in an amount between 250 and 750 μg, for example 500 μg. 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 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, e.g. macrogol 15 hydroxystearic acid a first vial having ii. a second vial containing at least one buffering agent, preferably in dry form; and iii. Optionally, an accessory cartridge for eluting the radioisotope generated by the radioisotope generator. 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, an accessory cartridge for eluting the radioisotope generated by the radioisotope generator. 21. A kit for carrying out the method of embodiment 20, comprising: 37. The kit of embodiment 35 or 36, wherein the NeoB compound is contained in an amount of between 20 and 60 μg, typically 50 μg. 38. The kit of any one of embodiments 35-37, wherein gentisic acid is included in an amount between 50 and 250 μg, typically 200 μg. 39. The kit of any one of embodiments 35-38, wherein the bulking agent is mannitol in an amount between 10 and 30 mg, for example 20 mg. 40. The kit of any one of embodiments 35-39, wherein the surfactant is macrogol 15 hydroxystearic acid in an amount between 250 and 750 μg, for example 500 μg. 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 hydroxystearic acid The kit of any one of embodiments 35 to 40, comprising: 42. The kit of any one of embodiments 35-41, wherein the second vial or single vial contains a buffer to maintain a pH between 3.0 and 4.0. 43. The kit of any one of embodiments 35-42, wherein the second vial contains formic acid and sodium hydroxide as buffering agents. 44. The kit of any one of embodiments 35-43, wherein all components of the first, second, or single vial are in dry form. EXAMPLES

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

[0108] Radiochemical Purity by ITLC Preparation of mobile phase solutions: Ammonium acetate 5M: Accurately weigh 3.85 g (3.84615 ÷ 3.85385 g) of ammonium acetate into a 10 mL graduated flask and dissolve the ammonium acetate in 10 mL of MilliQ water.

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

[0110] Prepare ITLC-SA: Cut one piece of ITLC-SA for each vial to 115 mm, draw a line 20 mm from the bottom (where to place a 5 uL drop of sample) and draw a line 100 mm from the bottom (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 non-complexed species is68 Ga colloidal species and free form 68 (Refers to Ga).

[0111] Radiochemical purity by HPLC

[0112] [Table 1]

[0113] Example 1: Development of a method for radiolabeling NeoB with 68Gallium using a 2-vial kit 1.2- Description and composition of vial kit Applicant has developed a sterile 2-vial kit consisting of: Vial 1: 68 Ge / 68 Gallium chloride-68( 68 NeoB, 50 μg, powder for injection solution, dissolved in a solution of GaCl3); Vial 2: Reaction buffer.

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

[0115] One accessory cartridge is used to reduce the amount of germanium-68 (68Ge) ions potentially present in the generator eluate.

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

[0117] Equivalent to the amount of radioactivity administered, 68The volume of Ga-NeoB injection solution is calculated according to the estimated time of injection based on the current radioactivity provided by the generator and the physical decay of the radionuclide (half-life=68 min). 68 The Ga-NeoB injection solution is a single dose product.

[0118] Vial 1 is a powder for injection solution containing 50 μg NeoB as the active ingredient packaged in a 10 mL glass vial.

[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) is part of a radiopharmaceutical kit that also contains reaction buffer (vial 2) and an accessory cartridge.

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

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

[0126] 2.1.1 Drug Substances The active ingredient is the NeoB peptide, a seven amino acid long sequence covalently linked to a chelating agent (DOTA) by a PABZA-DIG linker, as shown in formula (I) below. [ka]

[0127] 2.1.2 Additives The excipients selected for the composition of vial 1 are selected to maintain the stability of the active ingredient in the final drug, to ensure the safety and efficacy of the formulation, and further, to ensure the stability of the active ingredient in the final drug, during the dissolution procedure. 68 In order to obtain the required radiochemical purity of the Ga-NeoB solution, it is added. The selected additives result in a formulation with the required drug technical characteristics.

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

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

[0130] Mannitol Mannitol is used as a bulking agent. Since peptide drugs are very potent, only very small amounts are required in the formulation. In the absence of a bulking agent, the processing of the product is not feasible from a technical point of view. The bulking agent allows the processing of the pharmaceutical product and the production of a proper lyophilized product.

[0131] Gentisic acid Gentisic acid is a non-pharmacopoeial excipient used as an antioxidant in pharmaceutical formulations.

[0132] Kolliphor HS 15 (Macrogol 15 Hydroxystearate) Kolliphor HS 15 is a water-soluble non-ionic solubilizer used in parenteral pharmaceuticals. As a solubilizer, Kolliphor HS 15 is particularly suitable for parenteral and oral dosage forms.

[0133] Due to the non-specific binding of the peptides used as active ingredients in the NeoB radiopharmaceutical kit, Kolliphor HS 15 is used as a surfactant for peptides that have a tendency to stick to glass and plastic surfaces. As a non-ionic surfactant, 68 There is no risk of interference during labeling with Ga.

[0134] 2.2 Preparation 2.2.1 Drug development The drug development is commercially available without any treatment of the eluate or any additional purification steps. 68 Ge / 68 This was carried out with the aim of identifying the composition of a reaction mixture that could allow simple labelling of DOTA-peptides based on their direct dissolution with the eluate produced by a Ga generator.

[0135] The goal was to develop a bombesin-like peptide antagonist (NeoB) for use as a radiotracer for the detection of GRPR-positive tumors.

[0136] Vial 1 during the radiolabeling procedure 68 It is a lyophilized powder containing a peptide as the active ingredient that is radiolabeled with Ga.

[0137] Initial attempts to develop a suitable drug for NeoB (vial 1) involved testing on a bulk solution prepared on a laboratory scale, prior to the sterilization and lyophilization process.

[0138] The development work has targeted radiochemical purity as follows: 68 To obtain the final product leading to the 1Ga radiolabeled NeoB product, we focused on the selection of appropriate additives with respect to the peptide characteristics. ·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, beginning with the amount of active ingredient and the selection of suitable excipients.

[0142] 2.2.1.1 Selection of peptide amount 1850MBq 68 Ge / 68 Using eluates produced by a Ga generator and formic acid buffer, increasing amounts of NeoB peptide (from 15 μg to 100 μg) were purified to greater than 98% by HPLC and 97% by ITLC. 68 The labeling procedure was tested to identify the minimum amount of peptide required to have Ga incorporation. Based on the results summarized in Table 5, the lowest amount of peptide that reproducibly yields good radiochemical purity is 25 μg.

[0143] [Table 5]

[0144] In parallel, different peptide doses were also tested in in vivo biodistribution experiments. Briefly, a prostate cancer model in mice was used to compare two doses with different peptide masses: 10 pmol vs. 200 pmol; the amount of total radioactivity injected was kept constant in these experiments (1 MBq). Injection of radiolabeled NeoB resulted in increased accumulation in the tumor when a higher peptide mass dose was used (200 pmol). At the same time, uptake in non-target organs (such as the pancreas) was significantly lower at the higher peptide mass dose (200 pmol). These preclinical evaluations therefore demonstrated that a higher peptide mass dose would be preferred, as it was associated with a reduced uptake in non-target organs (notably the pancreas in this case).

[0145] Based on the radiolabeling studies performed (described in Table 5) and in vivo biodistribution experiments showing that a higher peptide mass dose ensures better efficacy and safety profile of the compound, the final amount of peptide selected to be included in Vial 1 was 50 μg.

[0146] The drug development work also focused on the selection of surfactants, antioxidants, and bulking agents. Radiolabeling procedures were also carefully evaluated.

[0147] 2.2.1.2 Selection of important additives ·Surfactant selection During the tests carried out to determine the drug in vial 1 (NeoB 50 μg, powder for injection solution), the peptide appeared to have a certain tendency to stick to glass and plastic surfaces. This phenomenon is called non-specific binding (NSB). Peptides often present a greater NSB problem than small molecules, especially uncharged peptides, which can stick strongly to plastic. The causes may be various: physical / chemical properties, van der Waals interactions, ionic interactions. Therefore, an evaluation was made on the addition of additives known to reduce NSB, including surfactants and solubilizers.

[0148] Organic solvents may enhance solubility and prevent adsorption. Ethanol can be used, for example, in radiopharmaceutical injections to enhance the solubility of highly lipophilic tracers or to reduce adsorption to vials, membrane filters, and syringes. Ethanol is not an option for NeoB powder for injection solutions, as it is not compatible with the freeze-drying process.

[0149] Human serum albumin (HSA) is also used in many protein drugs as a stabilizer to prevent surface adsorption, but this additive is not suitable due to its thermal instability.

[0150] Another possible approach to reduce peptide non-specific binding has been the use of detergents (e.g., polysorbate 20, polysorbate 80, Pluronic F-68, sorbitan trioleate). Ionic detergents 68 Particular attention was paid to the investigation of non-ionic detergents, as they may interfere with Ga labeling.

[0151] Non-ionic surfactants such as Kolliphor HS 15, Kolliphor K188, Tween 20, Tween 80, Polyvinylpyrrolidone K10 are commercially available as solubilizing additives in oral and injectable drugs.

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

[0153] Hydroxypropyl β-cyclodextrin was also evaluated in the formulation, either alone or in combination with surfactant. As reported below, the presence of hydroxypropyl β-cyclodextrin had only a limited positive effect on peptide attachment. Furthermore, as demonstrated in subsequent studies (see also radiolabeling procedure in section 2.2.1.3), the presence of surfactant and hydroxypropyl β-cyclodextrin did not improve the radiochemical purity of the final product when compared to a formulation containing surfactant only. For this reason, hydroxypropyl β-cyclodextrin was not included in the final formulation.

[0154] [Table 6]

[0155] The best results in terms of peptide attachment were obtained with Kolliphor HS 15 and with Tween 20. The two additives were further investigated to determine the final amounts to be included in the kit. The results obtained were good in terms of radiochemical purity and peptide attachment.

[0156] [Table 7]

[0157] Kolliphor HS 15 was finally chosen because polysorbate (tween 20) may undergo autoxidation, cleavage at the ethylene oxide subunit, and hydrolysis of fatty acid ester bonds induced by the presence of oxygen, metal ions, peroxides, or elevated temperature.

[0158] The lowest peptide deposition was obtained when using 0.5 mg of Kolliphor HS 15, which was the amount of Kolliphor HS 15 selected in the final composition of the formulation.

[0159] ·Selection of antioxidants The presence of the radical scavenger makes it possible to protect NeoB from radiolysis due to its antioxidant properties.

[0160] For development studies, the inventors investigated gentisic acid and ascorbic acid as antioxidants for use in radiopharmaceutical preparations. Tests were conducted to identify the lowest amount of antioxidant that could exert the desired protective function without interfering with the radiolabel.

[0161] Radiolabeling was primarily performed to identify the most suitable antioxidants and DOTA-peptides. 68 Varying amounts of antioxidants were tested while keeping other parameters constant to identify concentrations that would not interfere with Ga incorporation. As shown in the table below, gentisic acid: 68 It was identified as the best antioxidant since it does not interfere with Ga incorporation and is greater than 98% by HPLC. The amount of gentisic acid selected is 200 μg.

[0162] [Table 8]

[0163] [Table 9]

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

[0165] Among the bulking agents commonly proposed for lyophilization of peptides, formulators have tested inositol and mannitol.

[0166] [Table 10]

[0167] Mannitol was chosen because it is the most commonly used in lyophilisates and is known to produce cakes with good characteristics in terms of appearance, stability and moisture in the freeze-drying process. Furthermore, mannitol has been described in the literature 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. In a heat block (make sure the temperature has already reached 95°C before starting the elution) 68 Ge / 68 in HCl provided by a Ga generator 68 The lyophilized drug (vial 1) is dissolved directly 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 heat for more than 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 study was conducted to identify values ​​that would show good incorporation in a time frame compatible with the short half-life of Ga.

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

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

[0174] The formulation tested in these initial studies included a solubilizing agent (hydroxypropyl beta-cyclodextrin). However, later during development, similar studies were performed with the same formulation but without hydroxypropyl beta-cyclodextrin, resulting in good radiochemical and chemical purity. In addition, peptide attachment was also shown to be unaffected by the absence of hydroxypropyl beta-cyclodextrin, and therefore hydroxypropyl beta-cyclodextrin was not included in the final formulation. At 80°C and 85°C, radiometric analysis showed sufficient incorporation in 7 minutes.

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

[0176] Based on these observations, we showed that 95°C for 7 min is the most conservative labeling condition and can guarantee >98% incorporation without significant fragmentation even when the temperature fluctuates within a range of ±15°C.

[0177] [Table 11]

[0178] Moreover, to increase the robustness of the labeling procedure, the addition of reaction buffer (vial 2) at room temperature (RT) was evaluated (only after adding reaction buffer, the labeling reaction was carried out at 95° C.). The results shown in Table 11 confirm that good radiochemical purity is obtained even under these conditions.

[0179] [Table 12]

[0180] 2.2.1.4 Final selected drug (vial 1) Based on all the development studies described above, the final composition of 50 μg NeoB, Powder for Injection Solution (Vial 1) will be as follows:

[0181] [Table 13]

[0182] The final drug was tested for radiolabeled product to confirm the results obtained during development.

[0183] [Table 14]

[0184] As shown in Table 13, good radiochemical purity results by both ITLC and HPLC (>92%) were obtained after three independent radiolabeling studies performed with the final drug. It is also important to note that free gallium (by HPLC) was always below 2%. Finally, peptide adhesion to glass was also tested during these radiolabeling studies, and the presence of Kolliphor HS15 was confirmed to be necessary to maintain peptide adhesion at acceptable levels.

[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 label pH is 68 Due to its specific chemical behavior, the radiolabeling rate of DOTA-peptides with GaCl3 is one of the important parameters for obtaining good results. In order to define the pH range in which the labeling gives good results, 68Gallium-labeled NeoB agents were tested while maintaining a pH range between 3.0 and 4.0. Labeling was tested while varying the volume of reaction buffer added and keeping other parameters constant. As shown in Tables 14 and 15, variation in pH within the range 3.0-4.0 does not affect the success of labeling. The radiolabeled products obtained meet the criteria for radiochemical purity.

[0187] [Table 15]

[0188] [Table 16]

[0189] Gentisic acid vs. volume radioactivity The test is 68 Ge / 68 The highest volumetric radioactivity that the Ga generator could provide at the time 68 The effect of gentisic acid as a radiolytic scavenger was evaluated when labeling was performed with GaCl. A fractional elution was performed with the highest possible volumetric radioactivity; only the fraction with the highest radioactivity was used for labeling.

[0190] The protective effect was confirmed by monitoring peptide fragmentation over a period of time in the presence of various amounts of gentisic acid (0.20 mg and 0.35 mg). The results (see Table 16) confirmed almost the same positive effect for both tests. Therefore, the lowest amount of gentisic acid (200 μg) sufficient to achieve a good level of protection from radiolysis was chosen.

[0191] [Table 17]

[0192] In addition, to test whether a lower amount of gentisic acid can still act as an antioxidant in the final drug, the first test was carried out using 0.1 mg of gentisic acid. The results of the radiolabeling test carried out under these conditions are shown in Table 17, which confirm that good radiochemical purity can be obtained even in the presence of a lower amount of gentisic acid. Nevertheless, the amount of gentisic acid in the final drug was kept at 200 μg as a precaution, in order to ensure that good radiochemical purity can be obtained due to the higher radioactivity of the generator.

[0193] [Table 18]

[0194] Scale-up batch - 68 Test results for Ga-radiolabelled products Table 18 summarizes two radiolabeling studies performed with scale-up batch NeoB vial 1. The results show the radiolabeled formulation obtained with the scale-up batch of vial 1. 68 Ga-NeoB was shown to meet radiochemical purity criteria up to 4 hours after completion of the radiolabeling reaction.

[0195] [Table 19]

[0196] References 1.Sah BR,Burger IA,Schibli R,Friebe M,Dinkelborg L,Graham K,Borkowski S,Bacher-Stier C,Valencia R,Srinivasan A et al:Dosimetry and First Clinical Evaluation of the New 18F-Radiolabeled Bombesin Analogue BAY 864367 in Patients with Prostate Cancer.J Nucl Med 2015,56(3):372-378. 2.Kahkonen E,Jambor I,Kemppainen J,Lehtio K,Gronroos TJ,Kuisma A,Luoto P,Sipila HJ,Tolvanen T,Alanen K et al:In vivo imaging of prostate cancer using[68Ga]-labeled bombesin analog BAY86-7548.Clin Cancer 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 clinical experience with the gastrin-releasing peptide receptor-antagonist[(68)Ga]SB3 and PET / CT.Eur J Nucl Med Mol Imaging 2016,43(5):964-973. 4. Dimitrakopoulou-Strauss A, Hohenberger P, Haberkorn U, Macke HR, Eisenhut M, Strauss LG: 68Ga-labeled bombesin studies in patients with gastrointestinal stromal tumors: comparison with 18F-FDG. J Nucl Med 2007, 48(8): 1245-1250. 5. Velikyan I, Xu H, Nair M, Hall H: Robust labeling and comparative preclinical characterization of DOTA-TOC and DOTA-TATE. Nucl Med Biol 2012, 39(5): 628-639. The present invention includes the following aspects. [1] A radioisotope, preferably 68 Ga, 67 Ga, or 64 1. A method for labeling a Gastrin Releasing Peptide Receptor (GRPR) antagonist with Cu, comprising: i. Providing a first vial containing said GRPR antagonist in a dried form. ii. adding a solution of said radioisotope into said first vial, thereby obtaining a solution of said GRPR antagonist carrying said radioisotope; iii. mixing the solution obtained in ii with at least one buffer and incubating it for a period of time sufficient to obtain said radiolabeled GRPR antagonist; and iv. Optionally, adjusting the pH of the solution. The method includes: [2] The method according to [1], wherein the first vial in step i. is a reaction vial containing the GRPR antagonist and a buffer, preferably both in dry form. [3] The method according to [1], wherein step iii comprises mixing the solution obtained in ii with at least one reaction solution containing a buffer and incubating it for a period of time sufficient to obtain the GRPR antagonist labeled with the radioisotope. [4] The GRPR antagonist has formula (I):

Chem.

[10]

[0023] For use as an injection for in vivo detection of tumors by imaging diagnosis in a subject in need thereof, the injection can be obtained or obtained by any one of the methods of [4] to [8]. 68 A solution containing a compound of formula (I) according to [4], which is labeled with Ga.

[11] Consists of the following components in dry form: i. The 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 selected from the group consisting of the amino acid residues Asn, Thr, Phe, 3-(2-thienyl)alanine (Thi), 4-chlorophenylalanine (Cpa), α-naphthylalanine (α-Nal), β-naphthylalanine (β-Nal), 1,2,3,4-tetrahydronorharman-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 selected from -NHOH, -NHNH2, -NH-alkyl, -N(alkyl)2, and -O-alkyl; or Z is

change

[12] The GRPR antagonist has formula (I):

change

[11] , which is a compound of the formula:

[13] Consists of: - an amount between 20 and 60 μg, typically 50 μg of the following formula (I);

change

[11] to

[12] ,

[14] 1. A kit comprising: i. The following components in dry form: a compound of formula (I):

change

[15] A kit comprising: i. The following components in dry form: a compound of formula (I):

change

[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);

change

[14] to

[15] ,

[17] The kit according to any one of

[14] to

[16] , wherein all components of the first, second, or single vial are in a dried form.

Claims

1. By radioisotope, 【Chemistry 1】 1. A method for labeling a gastrin releasing peptide receptor (GRPR) antagonist of i. Providing a first vial containing the GRPR antagonist in a dry form. ii. adding a solution of the radioisotope into the first vial, thereby obtaining a solution of the GRPR antagonist with the radioisotope; iii. Mixing the solution obtained in ii with at least one first buffer and incubating it for a period of time sufficient to obtain the radioisotope-labeled GRPR antagonist. Including, The method, wherein the first vial further comprises macrogol 15 hydroxystearate or polysorbate 20.

2. The radioisotope ６８ G.A. ６７ Ga, or ６４ The method of claim 1 , wherein the metal is Cu.

3. 2. The method of claim 1, wherein the first vial in step i. is a reaction vial containing the GRPR antagonist and a second buffering agent.

4. 2. The method of claim 1, wherein step iii comprises mixing the solution obtained in ii with at least one reaction solution comprising a first buffer and incubating it for a period of time sufficient to obtain the radioisotope-labeled GRPR antagonist.

5. The method of claim 1, further comprising step (iv) of adjusting the pH of the solution.

6. The method of any one of claims 1 to 5, wherein the GRPR antagonist is contained in the first vial in an amount of between 20 and 60 μg.

7. The method of any one of claims 1 to 6, wherein the first vial further comprises gentisic acid as a radiolysis protectant.

8. 8. The method of claim 7, wherein gentisic acid is present in an amount between 50 and 250 μg.

9. The method of any one of claims 1 to 8, wherein the first vial further comprises mannitol.

10. The following components in dry form: i. Formula (I): 【Chemistry 2】 ii. Radiolytic protective substances; iii. Bulking agents; and iv. Macrogol 15 hydroxystearate or polysorbate 20 4. A powder for injection solution comprising:

11. Consists of: - between 20 and 60 μg of the following formula (I); 【Chemistry 3】 - Gentisic acid in an amount between 50 and 250 μg, - mannitol in an amount between 10 and 30 mg; - Macrogol 15 hydroxystearic acid in an amount between 250 and 750 μg 11. A powder for injection solutions according to claim 10, comprising:

12. A powder for injection solution as described in claim 11 for use in the preparation of an injection solution labeled with 68 Ga for in vivo detection of tumors by diagnostic imaging in a subject in need thereof.

13. 1. A kit comprising: i. The following components in dry form: a compound of formula (I): 【Chemistry 4】 - radiolytic protective substances, - bulking agents, and - Macrogol 15 hydroxystearate or polysorbate 20 a first vial having ii. a second vial containing at least one buffering agent; Including the kit.

14. A kit comprising: i. The following components in dry form: a compound of formula (I): 【Chemistry 5】 - radiolytic protective substances, - bulking agents, macrogol 15 hydroxystearate or polysorbate 20; and at least one buffer A single vial having Including the kit.

15. The first or single vial contains the following components: - an amount of between 20 and 60 μg of the following compounds of formula (I); 【Chemistry 6】 - gentisic acid in an amount between 50 and 250 μg, - mannitol in an amount between 10 and 30 mg, and The kit according to claim 13 or 14, comprising macrogol 15 hydroxystearic acid in an amount between -250 and 750 μg.

16. The kit according to any one of claims 13 to 15, wherein all components of the first, second or single vial are in dry form.

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