Stable concentrated radiopharmaceutical compositions

Abstract

The present disclosure relates to highly concentrated and chemically stable solutions of radionuclide complexes that can be used as pharmaceuticals for diagnostic and / or therapeutic purposes. The stability of the pharmaceuticals is achieved by at least one stabilizer against radiolysis. The use of two stabilizers introduced at different stages during the manufacturing process has been found to be particularly advantageous.

Description

[Technical Field]

[0001] The present disclosure relates to pharmaceutical compositions comprising radiolabeled GRPR antagonist compounds at high concentrations and with high chemical and radiochemical stability, allowing for their use as commercial pharmaceuticals for diagnostic and / or therapeutic purposes. [Background technology]

[0002] Bombesin was first isolated from the European frog Bombina bombina and was demonstrated to mimic mammalian gastrin-releasing peptide (GRP) and neuromedin B (NMB) [Scopinaro F, et al. Eur J Nucl Med Mol Imaging 2003,30(10):1378-1382].

[0003] Gastrin-releasing peptide (GRP), a bombesin-like peptide growth factor, regulates numerous functions in the gastrointestinal and central nervous systems, including the release of gut hormones, smooth muscle cell contraction, and epithelial cell proliferation. GRP is a potent mitogen for physiological and neoplastic tissues and may be involved in growth dysregulation and carcinogenesis.

[0004] The actions 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 various brain and neuronal tumors. In breast cancer, overexpression of GRPR can reach extremely high densities depending on the tumor type (e.g., 70-90% expression in ductal carcinoma specimens) [Van de Wiele C, et al. J Nucl Med 2001, 42(11):1722-1727].

[0005] GRPR is highly overexpressed in prostate cancer, where it has been shown to have both high affinity (nM levels) and high tumor uptake (%ID / g) in studies in human prostate cancer cell lines and xenograft models, although the relative expression of GRPR across the evolving disease landscape from early to late has yet to be 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 have been determined by immunohistochemistry in randomly selected colon cancer specimens, including LN and metastatic lesions. More than 80% of specimens abnormally expressed either GRP or GRPR, and more than 60% expressed both GRP and GRPR, but expression was not observed in adjacent normal healthy epithelium [Scopinaro F, et al. Cancer Biother Radiopharm 2002,17(3):327-335].

[0007] GRP is physiologically present in pulmonary neuroendocrine cells and plays a role in stimulating lung development and maturation. However, GRP also appears to be involved in proliferation dysregulation and carcinogenesis. GRP stimulation increases the release of epidermal growth factor receptor (EGFR) ligands, subsequently activating EGFR and downstream mitogen-activated protein kinase pathways. Using non-small cell lung cancer (NSCLC) cell lines, it has been confirmed that both EGF and GRP stimulate NSCLC proliferation, and that inhibition of either EGFR or GRP causes 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 has been that internalization of the receptor-radiolind complex can result in high accumulation of radioactivity within target cells. In the case of radionuclide-labeled peptides, efficient receptor-mediated endocytosis in response to agonist stimulation leads to high in vivo radioactivity uptake in target tissues, a crucial prerequisite for optimal imaging of malignant tumors. However, a paradigm shift occurred when receptor-selective peptide antagonists demonstrated favorable biodistribution, including significantly greater in vivo tumor uptake compared to highly potent agonists. A further advantage offered by GRPR antagonists is safer clinical use, less so in terms of current diagnostic tracer doses, but more so in terms of larger doses for potential therapeutic purposes, since no acute adverse biological effects are expected from their use [Stoykow C, et al. Theranostics 2016,6(10):1641-1650].

[0009] Recently, it has become clear that some GRPR antagonists, such as NeoB, can be radiolabeled with various radionuclides and potentially used in imaging and treating GRPR-expressing cancers, including, but not limited to, prostate and breast cancer.

[0010] In non-clinical models, 68 Ga]-NeoB and [ 177 Lu]-NeoB showed high affinity for GRPR expressed in breast, prostate, and gastrointestinal stromal tumors (GISTs), as well as low internalization upon binding to the specific receptor. The ability of the radiolabeled peptide to target GRPR-expressing tumors was confirmed by in vivo imaging and biodistribution studies in animal models [Dalm et al. Journal of nuclear medicine 2017, Vol. 58(2): 293-299].

[0011] In this radiopharmaceutical application, the target cell receptor binding moiety is typically linked to a chelator capable of forming a strong complex with the metal ion of the radionuclide. The radiopharmaceutical is then delivered to the target cell, and decay of the radionuclide subsequently releases high-energy electrons, positrons, or alpha particles and gamma rays at the target site.

[0012] One of the technical problems with these radiopharmaceuticals is that radionuclide decay occurs constantly (e.g., during pharmaceutical manufacturing and storage), and the released high-energy emissions induce cleavage of chemical bonds in molecules that form part of the pharmaceutical. This is often referred to as radiolysis or radiolytic degradation. Radiolysis of the receptor-binding portion of the pharmaceutical can lead to a decrease in the effectiveness of the pharmaceutical to act as a diagnostic and / or therapeutic agent.

[0013] Because these radiopharmaceuticals are not very stable and do not have a meaningful shelf life, they have until now required to be manufactured as individual patient doses in hospital drug laboratories and administered immediately to patients already awaiting radiation treatment and who need to be at the hospital.

[0014] To reduce radiolysis of radiopharmaceuticals and thereby improve their stability, various strategies have been explored with more or less success: pharmaceuticals can be stored at low temperatures, or produced at high dilutions, or stabilizers can be added.

[0015] However, the addition of stabilizers can be problematic because these chemicals may adversely affect the complexation of the radionuclide to the chelating agent or may become solubility-limiting and precipitate out of solution. Ethanol has been reported as a stabilizer against radiolysis (WO 2008 / 009444). While ethanol may not adversely affect complexation or solubility issues, high amounts of ethanol in infusion solutions may be physiologically problematic and may adversely affect the tolerability of the drug.

[0016] Producing pharmaceuticals at high dilutions has the disadvantage of requiring large volumes of injection solution to be administered to patients. For reasons of patient convenience and drug tolerability, it would be highly desirable to provide radiopharmaceuticals at high concentrations. However, these highly concentrated solutions are particularly susceptible to radiolysis. Therefore, a contradictory situation exists: on the one hand, avoiding radiolysis by diluting the pharmaceutical, while on the other hand, avoiding patient discomfort during treatment by providing a concentrated drug solution. Mathur et al., Cancer Biotherapy and Radiopharmaceuticals, 2017, 32(7), 266-273, reported a highly concentrated product and claimed it was ready for immediate use. However, the composition contains a large amount of ethanol, which may pose a tolerability issue.

[0017] Therefore, it remains a challenge to design a ready-to-use radiopharmaceutical composition that can be produced on a commercial scale, delivered as a sufficiently stable, highly concentrated sterile solution that allows for convenient injection of small volumes to patients, and has a physiologically well-tolerated composition (e.g., an ethanol-free composition).

[0018] The present inventors have now discovered a method for designing and producing highly concentrated radionuclide complex solutions that are extremely stable chemically and radiochemically when stored at ambient temperature or even at elevated temperatures for short periods of time, and as a result can be produced on a commercial scale and supplied as ready-to-use radiopharmaceuticals. Summary of the Invention

[0019] The present disclosure is provided in various aspects as generally outlined below: 1. (a) Below: (ai) a radionuclide; (aii) a GRP receptor peptide antagonist binding moiety linked to a chelator; and a complex formed by (b) at least two stabilizers against radiolysis; (c) optionally a surfactant.

[0020] 2. The radioactive nuclide is 111 In, 18 F, 211 At, 82 Rb, 123 I, 131 I, 133 mIn, 99 mTc, 94 mTc, 67 Ga, 66 Ga, 68 Ga, 52 Fe, 169 Er, 72 As, 97 Ru, 203 Pb, 212 Pb, 62 Cu, 64 Cu, 67 Cu, 186 Re, 188 Re, 86 Y, 90 Y, 51 Cr, 52 mmn, 157 Gd, 177 Lu, 161 Tb, 69 Yb, 175 Yb, 105 Rh, 166 Dy, 166 Ho, 153 Sm, 149 Pm, 151 Pm, 172 Tm, 121 Sn, 117mSn, 213 Bi, 212 Bi, 142 Pr, 143 Pr, 198 Au, 199 Au, 89 Zr, 225 Ac, 43 Sc, 44 Sc, and 47 Sc, preferably 111 In, 177 Lu, 225 Ac, and 68 Ga, more preferably 177 2. The pharmaceutical composition of embodiment 1, wherein the compound is Lu.

[0021] 3. The pharmaceutical composition of embodiment 1, wherein said radionuclide is present in a concentration that results in a volumetric activity of at least 370 MBq / mL (at EOP) ± 37 MBq / mL (± 10%).

[0022] 4. The pharmaceutical composition of embodiment 1, wherein the chelating agent is selected from DOTA, DTPA, NTA, EDTA, DO3A, NOC, and NOTA, preferably DOTA.

[0023] 5. The pharmaceutical composition of embodiment 1, wherein the GRP receptor peptide antagonist binding moiety linked to said chelator is NeoB of formula (I). [ka]

[0024] 6. The pharmaceutical composition of embodiment 1, wherein the at least two stabilizers are selected from gentisic acid (2,5-dihydroxybenzoic acid) or a salt thereof, ascorbic acid (L-ascorbic acid, vitamin C) or a salt thereof (e.g., sodium ascorbate), methionine, histidine, melatonine, ethanol, and Se-methionine, preferably selected from gentisic acid or a salt thereof and ascorbic acid or a salt thereof.

[0025] 7. The pharmaceutical composition of embodiment 6, wherein the at least two stabilizers are gentisic acid or a salt thereof and ascorbic acid or a salt thereof.

[0026] 8. The pharmaceutical composition according to embodiment 7, wherein the ratio between gentisic acid and ascorbic acid is 1:16 to 1:10, typically 1:15 to 1:10, for example 1:12 to 1:11.

[0027] 9. The pharmaceutical composition according to embodiment 7 or 8, wherein said gentisic acid or salt thereof is present in a concentration of at least 1000 μg / mL, such as from 1000 μg / mL to 1500 μg / mL.

[0028] 10. The pharmaceutical composition according to any one of embodiments 7 to 9, wherein the ascorbic acid or salt thereof is present in a concentration of at least 10000 μg / mL, preferably at least 12000 μg / mL, preferably at least 15000 μg / mL, for example 12000 to 18000 μg / mL.

[0029] 11. The pharmaceutical composition according to any one of embodiments 7 to 10, wherein the gentisic acid or a salt thereof is present at a concentration of at least 1000 μg / mL, for example, from 1000 μg / mL to 1500 μg / mL, and the ascorbic acid or a salt thereof is present at a concentration of 15000 μg / mL, for example, from 12000 to 18000 μg / mL.

[0030] 12. The pharmaceutical composition according to any one of embodiments 1 to 11, wherein said pharmaceutical preparation has a radiochemical purity of more than 95% for up to 72 hours, preferably more than 98% for up to 72 hours.

[0031] 13. The pharmaceutical composition according to any one of embodiments 1 to 12, wherein the surfactant is a non-ionic surfactant.

[0032] 14. The pharmaceutical composition of embodiment 13, wherein the nonionic surfactant is selected from macrogol 15 hydroxystearate, poloxamer, polysorbate 20, polysorbate 80, or polyvinylpyrrolidone with an average molecular weight of 10,000.

[0033] 15. The pharmaceutical composition according to embodiment 14, wherein the nonionic surfactant is macrogol 15 hydroxystearate.

[0034] 16. The pharmaceutical composition according to any one of embodiments 1 to 15, wherein said surfactant is present in a concentration of at least 5 μg / mL, preferably at least 25 μg / mL, more preferably at least 50 μg / mL.

[0035] 17. The pharmaceutical composition according to embodiment 16, wherein said surfactant is present in a concentration comprised between 5 μg / mL and 5000 μg / mL, preferably between 25 μg / mL and 2000 μg / mL, more preferably between 50 μg / mL and 1000 μg / mL.

[0036] 18. The pharmaceutical composition of embodiment 17, wherein the surfactant is present in a concentration of 100 μg / mL.

[0037] 19. (a) Below: (ai) radionuclide 177 Lutetium (Lu-177) and (aii) Formula (I): [ka] with NeoB and the complex formed by; (b) gentisic acid or its salts and ascorbic acid or its salts; (c) optionally, macrogol 15 hydroxystearate; and (d) optionally, at least one other pharmaceutically acceptable excipient.

[0038] 20. The pharmaceutical composition according to embodiment 19, wherein the at least one other pharmaceutically acceptable excipient is selected from a buffer, and / or a solvent, and / or a pH adjusting agent.

[0039] 21. The pharmaceutical composition according to embodiment 20, wherein the buffer is selected from acetate buffer, citrate buffer, and phosphate buffer, preferably acetate buffer.

[0040] 22. The pharmaceutical composition according to embodiment 20 or 21, wherein the solvent is water for injection.

[0041] 23. The pharmaceutical composition of embodiment 20, 21, or 22, wherein the pH adjuster is NaOH.

[0042] twenty four. (a) Below: (ai) radionuclide 177 Lutetium (Lu-177) and (aii) Formula (I): [ka] with NeoB and the complex formed by; (b) gentisic acid or its salts and ascorbic acid or its salts; (c) macrogol 15 hydroxystearate and; (d) acetate buffer; (e) Water for injection; (f) NaOH and; (g) DTPA, and a pharmaceutical composition comprising the same.

[0043] 25. The pharmaceutical composition according to any one of embodiments 1 to 24, wherein the pharmaceutical composition is an aqueous solution.

[0044] 26. The pharmaceutical composition according to any one of embodiments 1 to 25, wherein the pharmaceutical composition is an infusion solution.

[0045] 27. A pharmaceutical composition according to any one of embodiments 1 to 26 for use in the treatment or prevention of cancer, typically GRPR-positive cancer.

[0046] 28. The pharmaceutical composition of any one of embodiments 1 to 27, wherein the solution is produced in a commercial scale manufacturing, in particular in a batch size of at least 0.5 Ci.

[0047] 29. A pharmaceutical composition according to any one of embodiments 1 to 28, which is for commercial use.

[0048] 30. A process for producing the pharmaceutical composition as defined above, comprising: (1) A complex of a radionuclide and a GRP receptor peptide antagonist binding moiety linked to a chelator is (1.1) preparing an aqueous solution containing only a radionuclide and one stabilizer against radiolysis, which is gentisic acid or its salts; (1.2) preparing an aqueous solution comprising a GRP receptor peptide antagonist binding moiety linked to a chelator and, optionally, a surfactant; and (1.3) mixing the solutions obtained in steps (1.1) and (1.2), heating the resulting mixture, and optionally filtering the resulting solution; and forming a process step by; (2) The complex solution obtained in step (1) (2.1) preparing a dilute aqueous solution optionally containing only one stabilizer against radiolysis, which is ascorbic acid; and (2.2.) Mixing the complex solution obtained in step (1) with the diluted solution obtained in step (2.1) to obtain a final solution; and a process step of diluting by

[0049] 31. The solution from step (1.1) is 177 31. The process of embodiment 30, comprising LuCl3 and HCl.

[0050] 32. The process according to any one of embodiments 30 to 31, wherein the solution prepared in step (1.1) contains only one stabilizer, which is gentisic acid or a salt thereof, at a concentration of at least 1000 μg / mL, for example, from 1000 μg / mL to 1500 μg / mL.

[0051] 33. The process according to any one of embodiments 30 to 32, wherein the solution of step (1.1) further comprises a buffer, preferably an acetate buffer.

[0052] 34. The process of any one of embodiments 30-33, wherein the GRP receptor peptide antagonist binding moiety linked to a chelator in the solution of step (1.2) is NeoB of formula (I). [ka]

[0053] 35. The process of any one of embodiments 30-34, wherein the solution of step (1.2) further comprises a surfactant that is macrogol 15 hydroxystearate.

[0054] 36. The process according to any one of embodiments 30 to 35, wherein the solution prepared in step (2.1) contains only one stabilizer, which is ascorbic acid or a salt thereof, at a concentration of at least 10000 μg / mL, preferably at least 12000 μg / mL, preferably at least 15000 μg / mL, for example 12000 to 18000 μg / mL.

[0055] 37. The process according to any one of embodiments 30 to 36, wherein the solution of step (1.2) further comprises macrogol 15 hydroxystearate at a concentration comprised between 5 μg / mL and 5000 μg / mL, preferably between 25 μg / mL and 2000 μg / mL, more preferably between 50 μg / mL and 1000 μg / mL, and even more preferably at a concentration of 100 μg / L.

[0056] 38. The process of any one of embodiments 30 to 37, wherein in step (1.3), the resulting mixture is heated to a temperature of 70 to 99°C, preferably 90 to 98°C, for 1 to 59 minutes, preferably 2 to 15 minutes.

[0057] 39. The process according to any one of embodiments 30 to 38, wherein the complex obtained at the end of step (1.3) is further filtered at 0.20 μm.

[0058] 40. The process of any one of embodiments 30-39, wherein the solution of step (2.1) further comprises a sequestering agent that is diethylenetriaminepentaacetic acid (DTPA) or a salt thereof.

[0059] 41. The process of any one of embodiments 30-40, wherein the solution of step (2.1) further comprises a pH adjuster that is NaOH.

[0060] 42. The process of any one of embodiments 30-41, wherein the solution of step (2.1) further comprises water for injection.

[0061] 43. The process according to any one of embodiments 30 to 42, wherein (3) a process step of sterile filtering the solution obtained in step (2); (4) aseptically dispensing the filtered solution obtained in step (3) into unit-dose containers, wherein the radionuclide is present at a concentration that results in a volumetric activity of at least 370 MBq / mL (at EOP) ± 37 MBq / mL (± 10%).

[0062] 44. The process of any one of embodiments 30-43, wherein the dose unit containers of step (4) are stoppered vials sealed within a lead container.

[0063] 45. An aqueous pharmaceutical solution obtained by a process defined by any one of embodiments 30 to 44. DETAILED DESCRIPTION OF THE INVENTION

[0064] definition In the following, the meanings of terms used in this specification are defined.

[0065] The term "about" or "approximately" is used herein to mean that the following value may vary by ±20%, preferably ±10%, more preferably ±5%, even more preferably ±2%, and even more preferably ±1%.

[0066] Unless otherwise defined, "%" as used herein has the meaning of weight percent (wt%), also referred to as weight / weight percent (w / w%).

[0067] "Total concentration": the sum of one or more individual concentrations.

[0068] "Aqueous solution": a solution of one or more solutes in water.

[0069] "A complex formed by (ai) a radionuclide and (aii) a cell receptor-binding organic moiety linked to a chelating agent": The radionuclide metal ion forms a non-covalent bond with a functional group of the chelating agent, such as an amine or a carboxylic acid. The chelating agent has at least two such complexing functional groups so as to form a chelate complex.

[0070] "Buffer solution of pH 4 to 6.0": It may be an acetate buffer solution, a citrate buffer solution (e.g., citrate + HCl or citric acid + disodium hydrogen phosphate), or a phosphate buffer solution (e.g., sodium dihydrogen phosphate + disodium hydrogen phosphate), preferably the buffer solution is an acetate buffer solution, and preferably the acetate buffer solution is composed of acetic acid and sodium acetate.

[0071] "Sequestrant", a chelating agent suitable for complexing radionuclide metal ions, preferably DTPA: diethylenetriaminepentaacetic acid.

[0072] A "pH adjuster" is a chemical substance added to a solution to adjust the pH value of the solution and thereby achieve desired performance. pH control can be achieved by adding a pH adjuster to the formulation. Examples of pH adjusters include commonly used acids and bases, buffers, and mixtures of acids and bases. For example, bases that can be used include NaOH, KOH, Ca(OH), sodium bicarbonate, potassium carbonate, and sodium carbonate. Examples of acids that can be used include hydrochloric acid, acetic acid, citric acid, formic acid, fumaric acid, and sulfamic acid. Preferably, the pH adjuster is a base, more preferably NaOH. The pH of the fluid can range from about 2 to about 14, or any suitable range.

[0073] "For commercial use": A pharmaceutical product, e.g., an aqueous pharmaceutical solution, that can be (and preferably has been) approved for marketing by a health authority (e.g., the US FDA or EMA) by complying with all pharmaceutical quality and stability requirements required by such health authority, that can be (and preferably has been) manufactured on a commercial scale from or at a pharmaceutical production site, that has subsequently been subjected to quality control testing procedures, and that can be (and preferably is) supplied to end users (e.g., hospitals or patients) in remote locations.

[0074] Chelants relevant to the present disclosure include: DOTA: 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid; DTPA: diethylenetriaminepentaacetic acid; NTA: nitrilotriacetic acid; EDTA: ethylenediaminetetraacetic acid; DO3A: 1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid; NOTA: 1,4,7-triazacyclononane-1,4,7-triacetic acid; Trizoxetan, Tetraxetane Or it may be a mixture thereof, preferably DOTA.

[0075] "Cell receptor binding moiety": a chemical molecule, at least a portion of which binds to a receptor molecule on the surface of a cell. Cell receptor binding moieties particularly suited to the present disclosure are somatostatin receptor binding peptides, preferably the somatostatin receptor binding peptides are selected from octreotide, octreotate, lanreotide, vapreotide, pasireotide, iratreotide, pentetreotide, depreotide, satreotide, veldreotide, and preferably octreotide and octreotate.

[0076] "Linked": The cell receptor binding organic moiety is either directly linked to the chelator or connected via a linker molecule, preferably directly linked. The linking bond is either a covalent or non-covalent bond between the cell receptor binding organic moiety (and linker) and the chelator, preferably the bond is a covalent bond.

[0077] "Radiolytic stabilizer": a stabilizer that protects organic molecules from radiolysis. For example, when gamma rays emitted from a radionuclide break the bonds between atoms of an organic molecule and radicals are formed, these radicals are then trapped by a stabilizer that prevents the radicals from undergoing any other chemical reactions that may result in unwanted, potentially ineffective, or even toxic molecules. Therefore, these stabilizers are also called "free radical scavengers" or "radical scavengers" for short. Other terms for these stabilizers are "radiostability enhancers," "radiolytic stabilizers," or simply "quenchers."

[0078] "Radiochemical purity": The percentage of a stated radionuclide present in a stated 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.

[0079] As used herein, the term "effective amount" or "therapeutically effective amount" of a compound refers to an amount of compound that elicits a biological or medical response in a subject, e.g., ameliorates symptoms, alleviates a condition, slows or delays the progression of a disease, or prevents a disease.

[0080] As used herein, the term "substituted" or "optionally substituted" refers to any of a number of substituted or unsubstituted groups ranging from zero to the total number of open valences on the aromatic ring system, including 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) refers to a group optionally substituted with one or more substituents selected from 2R', -S(O)2NR'R'', -NRSO2R', -CN, -NO2, -R', -N3, -CH(Ph)2, fluoro(C1-C4)alkoxo, and fluoro(C1-C4)alkyl, where R', R'', R''', and R'''' can 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 of the R groups is independently selected as an R' group, an R'', an R''', and an R'''' group, respectively, when two or more of these groups are present.

[0081] As used herein, the term "alkyl," by itself 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, isopentyl), and hexyl and its isomers (e.g., n-hexyl, isohexyl).

[0082] As used herein, the term "heteroaryl" refers to a polyunsaturated aromatic ring system containing 5 to 10 atoms, having a single ring or multiple aromatic rings fused or covalently linked together, where 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 an aryl ring, a cycloalkyl ring, or a heterocyclyl ring. 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.

[0083] As used herein, the term "aryl" refers to a polyunsaturated aromatic hydrocarbyl group containing 6 to 10 ring atoms and having a single ring or multiple aromatic rings fused together, wherein at least one ring is aromatic. The aromatic ring may optionally contain one to two additional rings (cycloalkyl, heterocyclyl, or heteroaryl, as defined herein) fused thereto. Suitable aryl groups include phenyl, naphthyl, and phenyl rings fused to a heterocyclyl, such as benzopyranyl, benzodioxolyl, benzodioxanyl, and the like.

[0084] As used herein, the term "halogen" refers to a fluoro (-F), chloro (-Cl), bromo (-Br), or iodo (-I) group.

[0085] As used herein, the term "optionally substituted aliphatic chain" refers to an optionally substituted aliphatic chain having 4 to 36 carbon atoms, preferably 12 to 24 carbon atoms.

[0086] The present disclosure is further described and exemplified below.

[0087] As used herein, the term "ratio between gentisic acid and ascorbic acid" refers to the free acid concentration ratio (μg / mL:μg / mL), i.e., the concentration ratio for GA and AA as free acids, and does not include the concentration of counterions such as sodium (Na).

[0088] Generally, the present disclosure relates to pharmaceutical compositions, particularly radiopharmaceutical compositions. The pharmaceutical compositions are for intravenous (IV) use / application / administration. The solutions are stable, concentrated, and ready to use.

[0089] A radiopharmaceutical composition according to the present disclosure comprises: (a) Below: (ai) a radionuclide; (aii) a GRP receptor peptide antagonist binding moiety linked to a chelator; and a complex formed by (b) at least two stabilizers against radiolysis; (c) optionally a surfactant.

[0090] Radiolabeled GRPR antagonist The complex has the formula: MC-SP [In the formula, M is a radionuclide suitable for nuclear medicine, C is a chelator that binds to M; S is an optional spacer covalently attached between C and the N-terminus of P; P is a GRP receptor peptide antagonist, preferably having 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 as the L- or D-isomer; 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 [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 aryl or heteroaryl group substituted with alkyl-, halogen, hydroxyl, hydroxyalkyl, amine, amino, amido, or amide.

[0091] According to one embodiment, Z is selected from one of the following formulae (wherein X is NH or O): [ka]

[0092] According to one embodiment, P is DPhe-Gln-Trp-Ala-Val-Gly-His-Z; wherein Z is as defined above.

[0093] 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 or Z is [ka] wherein X is NH (amide), R2 is (CH2-CH(CH3)2, and R1 is (CH2N)-Pro-NH2, which is the same as or different from R2.

[0094] According to one embodiment, the chelator C is obtained by grafting one chelating agent selected from the following list: [ka]

[0095] According to one embodiment, M is 111 In, 18 F, 211 At, 82 Rb, 123 I, 131 I, 133 mIn, 99 mTc, 94 mTc, 67 Ga, 66 Ga, 68 Ga, 52 Fe, 169Er, 72 As, 97 Ru, 203 Pb, 212 Pb, 62 Cu, 64 Cu, 67 Cu, 186 Re, 188 Re, 86 Y, 90 Y, 51 Cr, 52 mmn, 157 Gd, 177 Lu, 161 Tb, 69 Yb, 175 Yb, 105 Rh, 166 Dy, 166 Ho, 153 Sm, 149 Pm, 151 Pm, 172 Tm, 121 Sn, 117 mSn, 213 Bi, 212 Bi, 142 Pr, 143 Pr, 198 Au, 199 Au, 89 Zr, 225 Ac, 43 Sc, 44 Sc, and 47 Preferably, M is a radionuclide suitable for nuclear medicine selected from: 111 In, 177 Lu, 225 Ac, and 68 Ga.

[0096] According to one embodiment, the chelator C is selected from the group consisting of DOTA, DTPA, NTA, EDTA, DO3A, NOC and NOTA, preferably DOTA.

[0097] According to one embodiment, S is a) Formula: [ka] an aryl-containing residue of the formula (wherein PABA is p-aminobenzoic acid, PABZA is p-aminobenzylamine, PDA is phenylenediamine, and PAMBZA is (aminomethyl)benzylamine); b) a dicarboxylic acid, an ω-aminocarboxylic acid, an ω-diaminocarboxylic acid, or a compound of the formula: [ka] diamines of the formula (where DIG is diglycolic acid and IDA is iminodiacetic acid); c) PEG spacers of various chain lengths, especially PEG spacer sele [ka] d) α- and β-amino acids in single chains or homologous chains of various lengths or heterologous chains of various 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.

[0098] According to one embodiment, the GRPR antagonist has the following formula: [ka] wherein MC and P are as defined above.

[0099] According to one embodiment, P is DPhe-Gln-Trp-Ala-Val-Gly-His-NH-CH(CH2-CH(CH3)2)2.

[0100] According to one embodiment, the complex has formula (I): [ka] NeoB1 ((DOTA-(p-aminobenzylamine-diglycolic acid)-[D-Phe-Gln-Trp-Ala-Val-Gly-His-NH-CH[CH2-CH(CH3)2]2).

[0101] According to one embodiment, the complex has the formula (II): [ka] Radiolabeled M-NeoB1 ((M-DOTA-(p-aminobenzylamine-diglycolic acid)-[D-Phe-Gln-Trp-Ala-Val-Gly-His-NH-CH[CH2-CH(CH3)2]2; wherein M is a radionuclide, preferably M is 177 Lu, 68 Ga and 111 In).

[0102] According to one embodiment, the radiolabeled GRPR antagonist has formula (III): [ka] Radiolabeled NeoB2 ((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.

[0103] In one embodiment, M is 111 In, 133 mIn, 99 mTc, 94 mTc, 67 Ga, 66 Ga, 68 Ga, 52 Fe, 169 Er, 72 As, 97 Ru,203 Pb, 212 Pb, 62 Cu, 64 Cu, 67 Cu, 186 Re, 188 Re, 86 Y, 90 Y, 51 Cr, 52 mmn, 157 Gd, 177 Lu, 161 Tb, 69 Yb, 175 Yb, 105 Rh, 166 Dy, 166 Ho, 153 Sm, 149 Pm, 151 Pm, 172 Tm, 121 Sn, 117 mSn, 213 Bi, 212 Bi, 142 Pr, 143 Pr, 198 Au, 199 Au, 89 Zr, 225 Ac, and 47 Preferably, M is a radionuclide that may be selected from: 111 In, 177 Lu, 225 Ac, and 68 Ga.

[0104] According to one embodiment, M is 177 In this case, the radiolabeled GRPR antagonist can be used in radionuclide therapy. According to another embodiment, M is 68 In this case, the radiolabeled GRPR antagonist can be used for PET. According to another embodiment, M is 111 In this case, the radiolabeled GRPR antagonist can be used for SPECT.

[0105] According to another particular embodiment, the GRPR antagonist has the following formula (IV): [ka] ProBOMB1 (DOTA-pABzA-DIG-D-Phe-Gln-Trp-Ala-Val-Gly-His-Leu-Ψ(CH2N)-Pro-NH2).

[0106] Synthesis of Compounds of Formulas (I), (II), (III), and (IV) Compounds of formula (I), (II), (III), and (IV) may 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.

[0107] Pharmaceutical Composition Radiolabeled GRPR antagonists tend to degrade over time, ultimately resulting in radiochemical purity below specification at the end of the target shelf life (72 hours). This presents a problem for the formulation of pharmaceutical compositions. Solution stability against radiolysis was confirmed by the use of stabilizers.

[0108] As used herein, "radiolytic stabilizers" refer to stabilizers that protect organic molecules from radiolysis; for example, when gamma rays emitted from radionuclides break bonds between atoms of organic molecules to form radicals, these radicals are then trapped by stabilizers that prevent the radicals from undergoing any other chemical reactions that may result in molecules that are undesirable, ineffective, or even toxic. Therefore, these stabilizers are also called "free radical scavengers" or "radical scavengers" for short. Other terms for these stabilizers are "radiostability enhancers," "radiolytic stabilizers," or simply "quenchers."

[0109] Generally, the stabilizer used according to the present invention may be selected from gentisic acid (2,5-dihydroxybenzoic acid) or its salts, ascorbic acid (L-ascorbic acid, vitamin C) or its salts (e.g., sodium ascorbate), methionine, histidine, melatonin, ethanol, and Se-methionine. Preferred stabilizers are selected from gentisic acid or its salts and ascorbic acid or its salts.

[0110] Ethanol is considered to be a less preferred stabilizer when present at high concentrations due to the associated tolerability problems.Ideally, ethanol should be avoided (in other words, ethanol-free) in the solution of the present disclosure, and at least, the amount of ethanol in the solution of the present disclosure should be limited to, for example, less than 5%, preferably less than 2%, more preferably less than 1% in the final solution to be injected / infused.Even more preferably, the solution does not contain ethanol.

[0111] In a first aspect, the present disclosure relates to a pharmaceutical composition comprising a radiolabeled GRPR antagonist as described herein and at least two stabilizers against radiolysis. In one embodiment, the at least two stabilizers may be selected from gentisic acid (2,5-dihydroxybenzoic acid) or a salt thereof, ascorbic acid (L-ascorbic acid, vitamin C) or a salt thereof (e.g., sodium ascorbate), methionine, histidine, melatonin, ethanol, and Se-methionine, and preferably from gentisic acid or a salt thereof and ascorbic acid or a salt thereof. The at least two stabilizers may be selected from gentisic acid or a salt thereof and ascorbic acid or a salt thereof.

[0112] In particular, the inventors have unexpectedly discovered that the addition of specific amounts of both ascorbic acid and gentisic acid to a pharmaceutical composition of a radiolabeled GRPR antagonist compound allows the composition to have a radiochemical purity of greater than 95% 72 hours after synthesis.

[0113] In one embodiment, the ratio between gentisic acid and ascorbic acid is 1:16 to 1:10, typically 1:15 to 1:10, for example 1:12 to 1:11.

[0114] In one embodiment, the gentisic acid or salt thereof may be present at a concentration of at least 1000 μg / mL, for example, 1000 μg / mL to 1500 μg / mL.

[0115] In one embodiment, the ascorbic acid or salt thereof may be present at a concentration of at least 10,000 μg / mL, preferably at least 12,000 μg / mL, preferably at least 15,000 μg / mL, for example, 12,000 to 18,000 μg / mL.

[0116] In one embodiment, the gentisic acid or a salt thereof is present at a concentration of at least 1000 μg / mL, for example, 1000 μg / mL to 1500 μg / mL, and the ascorbic acid or a salt thereof is present at a concentration of 15000 μg / mL, for example, 12000 to 18000 μg / mL.

[0117] In one embodiment, the radiopharmaceutical composition comprises both gentisic acid and ascorbic acid as radiation stabilizers at concentrations of 1312 μg / mL and 15000 μg / mL, respectively.

[0118] In one embodiment, the pharmaceutical composition has a radiochemical purity of greater than 95% for up to 72 hours, preferably greater than 98% for up to 72 hours.

[0119] GRPR antagonists tend to adhere to glass and plastic surfaces by non-specific binding (NSB), which is problematic for formulating pharmaceutical compositions.

[0120] In a second aspect, the present disclosure relates to a pharmaceutical composition comprising a radiolabeled GRPR antagonist as described herein, at least two stabilizers against radiolysis, and optionally a surfactant.

[0121] The surfactant may include a compound having (i) a polyethylene glycol chain and (ii) a fatty acid ester. In one embodiment, the surfactant also includes free ethylene glycol.

[0122] In one embodiment, the surfactant has the formula (V): [ka] (wherein n is 3 to 1000, preferably 5 to 500, more preferably 10 to 50, R is a fatty acid chain, preferably an optionally substituted aliphatic chain).

[0123] In one embodiment, the surfactant comprises polyethylene glycol 15-hydroxystearate and free ethylene glycol.

[0124] In one embodiment, the surfactant is a nonionic surfactant. Preferably, the nonionic surfactant is selected from macrogol 15 hydroxystearate (Kolliphor HS 15), poloxamer (Kolliphor P188), polysorbate 20 (Tween 20), polysorbate 80 (Tween 80), or polyvinylpyrrolidone having an average molecular weight of 10,000 (polyvinylpyrrolidone K10). Preferably, the nonionic surfactant is macrogol 15 hydroxystearate (Kolliphor HS 15).

[0125] The radiolabeled GRPR antagonist may be present at a concentration that results in a volumetric radioactivity of 370 MBq / mL (at EOP) ± 37 MBq / mL (± 10%).

[0126] The surfactant may be present at a concentration of at least 5 μg / mL, preferably at least 25 μg / mL, more preferably at least 50 μg / mL. The surfactant may be present at a concentration comprised between 5 μg / mL and 5000 μg / mL, preferably between 25 μg / mL and 2000 μg / mL, more preferably between 50 μg / mL and 1000 μg / mL. The surfactant may be present at a concentration of 100 μg / mL.

[0127] In a third aspect, the present disclosure relates to a pharmaceutical composition comprising a radiolabeled GRPR antagonist as described herein, at least two stabilizers against radiolysis, and optionally a surfactant and at least one other pharmaceutically acceptable excipient.

[0128] Pharmaceutically acceptable excipients can be any of those conventionally used, limited only by physicochemical considerations such as solubility and lack of reactivity with the active compound.

[0129] In particular, the one or more excipients may be selected from buffers, and / or solvents, and / or pH adjusters.

[0130] Buffers include acetate buffers, citrate buffers, and phosphate buffers, hi one embodiment, the buffer is an acetate buffer.

[0131] In one embodiment, the solvent is water for injection.

[0132] In one embodiment, the pH adjuster is NaOH.

[0133] In a fourth aspect, the present disclosure provides a compound comprising the radionuclide lutetium-177 (Lu-177) and a compound of formula (I): [ka] NeoB (DOTA-(p-aminobenzylamine-diglycolic acid)-[D-Phe-Gln-Trp-Ala-Val-Gly-His-NH-CH[CH2-CH(CH3)2]2) and a complex formed by The present invention relates to a pharmaceutical composition comprising gentisic acid or a salt thereof, ascorbic acid or a salt thereof, macrogol 15 hydroxystearate, acetate buffer, water for injection, and NaOH.

[0134] According to one embodiment, the pharmaceutical composition is an aqueous solution, such as an injectable formulation. According to a particular embodiment, the pharmaceutical composition is an infusion solution.

[0135] The requirements for effective pharmaceutical carriers for injectable compositions are well known to those skilled 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)).

[0136] The present disclosure also relates to the above-mentioned pharmaceutical composition for use in the treatment or prevention of cancer, typically GRPR-positive cancer.

[0137] As used herein, the term "cancer" refers to cells capable of autonomous growth (i.e., an abnormal state or condition characterized by rapidly proliferating cell proliferation). Hyperproliferative and neoplastic conditions may be classified as pathological (i.e., characterizing or constituting a pathological condition) or non-pathological (i.e., deviations from normal but not associated with a pathological condition). The term is intended to include all types of cancerous growths or oncogenic processes, metastatic tissues, or malignantly transformed cells, tissues, or organs, regardless of histopathological type or stage of invasiveness.

[0138] In certain embodiments, the cancer is selected from prostate cancer, breast cancer, small cell lung cancer, colon cancer, gastrointestinal stromal tumor, gastrinoma, renal cell carcinoma, gastrointestinal-pancreatic neuroendocrine tumor, esophageal squamous cell tumor, neuroblastoma, head and neck squamous cell carcinoma, as well as ovarian tumor, endometrial tumor, and pancreatic tumor, which exhibit neoplasia-associated vasculature that is GRPR. In one embodiment, the cancer is prostate cancer or breast cancer.

[0139] In another embodiment of the present invention, the pharmaceutical composition is produced in a commercial scale manufacturing, particularly in a batch size of at least 0.5 Ci.

[0140] In another aspect of the invention, the pharmaceutical composition is for commercial use.

[0141] In a further aspect, the present disclosure also provides a radiolabeled GRPR antagonist (typically a 177

[0023] For pharmaceutical compositions comprising Lu-NeoB, the pharmaceutical composition is formulated with a radiation stabilizer as described in any of the previous embodiments and administered to the subject in a therapeutically effective amount comprised between 2000 and 10000 MBq, and typically has a radiochemical purity (RCP) of greater than 95% upon administration.

[0142] In certain embodiments, the subject is a mammal, such as, but not limited to, a rodent, dog, cat, or primate. In preferred embodiments, the subject is a human.

[0143] In certain embodiments, a therapeutically effective amount of the composition is administered to the subject 2-8 times per treatment.

[0144] For example, radiolabeled GRPR antagonists (especially 177 Human patients can be treated by intravenously administering 2 to 8 cycles (2,000 to 10,000 MBq each) of the above pharmaceutical composition containing Lu-NeoB, typically with a radiochemical purity (RCP) of greater than 95% at the time of administration.

[0145] In accordance with the present disclosure, the following embodiments are provided: 1. (a) Below: (ai) a radionuclide; (aii) a GRP receptor peptide antagonist binding moiety linked to a chelator; and a complex formed by (b) at least two stabilizers against radiolysis; (c) optionally a surfactant.

[0146] 2. The radioactive nuclide is 111 In, 18 F, 211 At, 82 Rb, 123 I, 131 I, 133 mIn, 99 mTc, 94 mTc, 67 Ga, 66 Ga, 68 Ga, 52 Fe, 169 Er, 72 As, 97 Ru, 203 Pb, 212 Pb, 62 Cu, 64 Cu, 67 Cu, 186 Re, 188 Re, 86 Y, 90 Y, 51 Cr, 52 mmn, 157 Gd, 177 Lu, 161 Tb, 69 Yb, 175 Yb, 105 Rh, 166 Dy, 166 Ho, 153 Sm, 149 Pm, 151 Pm, 172 Tm, 121 Sn, 117 mSn, 213 Bi, 212 Bi, 142 Pr, 143 Pr,198 Au, 199 Au, 89 Zr, 225 Ac, 43 Sc, 44 Sc, and 47 Sc. Preferably, M is selected from: 111 In, 177 Lu, 225 Ac, and 68 Ga, more preferably 177 This is Lu.

[0147] 3. The pharmaceutical composition of embodiment 1, wherein said radionuclide is present in a concentration that results in a volumetric activity of at least 370 MBq / mL (at EOP) ± 37 MBq / mL (± 10%).

[0148] 4. The pharmaceutical composition of embodiment 1, wherein the chelating agent is selected from DOTA, DTPA, NTA, EDTA, DO3A, NOC, and NOTA, preferably DOTA.

[0149] 5. The pharmaceutical composition of embodiment 1, wherein the GRP receptor peptide antagonist binding moiety linked to said chelator is NeoB of formula (I). [ka]

[0150] 6. The pharmaceutical composition of embodiment 1, wherein the at least two stabilizers are selected from gentisic acid (2,5-dihydroxybenzoic acid) or a salt thereof, ascorbic acid (L-ascorbic acid, vitamin C) or a salt thereof (e.g., sodium ascorbate), methionine, histidine, melatonin, ethanol, and Se-methionine, preferably selected from gentisic acid or a salt thereof and ascorbic acid or a salt thereof.

[0151] 7. The pharmaceutical composition of embodiment 6, wherein the at least two stabilizers are gentisic acid or a salt thereof and ascorbic acid or a salt thereof.

[0152] 8. The pharmaceutical composition according to embodiment 7, wherein the ratio between gentisic acid and ascorbic acid is 1:16 to 1:10, typically 1:15 to 1:10, for example 1:12 to 1:11.

[0153] 9. The pharmaceutical composition according to embodiment 7 or 8, wherein said gentisic acid or salt thereof is present in a concentration of at least 1000 μg / mL, such as from 1000 μg / mL to 1500 μg / mL.

[0154] 10. The pharmaceutical composition according to any one of embodiments 7 to 9, wherein the ascorbic acid or salt thereof is present in a concentration of at least 10000 μg / mL, preferably at least 12000 μg / mL, preferably at least 15000 μg / mL, for example 12000 to 18000 μg / mL.

[0155] 11. The pharmaceutical composition according to any one of embodiments 7 to 10, wherein the gentisic acid or a salt thereof is present at a concentration of at least 1000 μg / mL, for example, from 1000 μg / mL to 1500 μg / mL, and the ascorbic acid or a salt thereof is present at a concentration of 15000 μg / mL, for example, from 12000 to 18000 μg / mL.

[0156] 12. The pharmaceutical composition according to any one of embodiments 1 to 11, wherein said pharmaceutical preparation has a radiochemical purity of more than 95% for up to 72 hours, preferably more than 98% for up to 72 hours.

[0157] 13. The pharmaceutical composition according to any one of embodiments 1 to 12, wherein the surfactant is a non-ionic surfactant.

[0158] 14. The pharmaceutical composition of embodiment 13, wherein the nonionic surfactant is selected from macrogol 15 hydroxystearate, poloxamer, polysorbate 20, polysorbate 80, or polyvinylpyrrolidone with an average molecular weight of 10,000.

[0159] 15. The pharmaceutical composition according to embodiment 14, wherein the nonionic surfactant is macrogol 15 hydroxystearate.

[0160] 16. The pharmaceutical composition according to any one of embodiments 1 to 15, wherein said surfactant is present in a concentration of at least 5 μg / mL, preferably at least 25 μg / mL, more preferably at least 50 μg / mL.

[0161] 17. The pharmaceutical composition according to embodiment 16, wherein said surfactant is present in a concentration comprised between 5 μg / mL and 5000 μg / mL, preferably between 25 μg / mL and 2000 μg / mL, more preferably between 50 μg / mL and 1000 μg / mL.

[0162] 18. The pharmaceutical composition of embodiment 17, wherein the surfactant is present in a concentration of 100 μg / mL.

[0163] 19. (b) Below: (ai) radionuclide 177 Lutetium (Lu-177) and (aii) Formula (I): [ka] with NeoB and the complex formed by; (b) gentisic acid or its salts and ascorbic acid or its salts; (c) optionally, macrogol 15 hydroxystearate; and (d) optionally, at least one other pharmaceutically acceptable excipient.

[0164] 20. The pharmaceutical composition according to embodiment 19, wherein the at least one other pharmaceutically acceptable excipient is selected from a buffer, and / or a solvent, and / or a pH adjusting agent.

[0165] 21. The pharmaceutical composition according to embodiment 20, wherein the buffer is selected from acetate buffer, citrate buffer, and phosphate buffer, preferably acetate buffer.

[0166] 22. The pharmaceutical composition according to embodiment 20 or 21, wherein the solvent is water for injection.

[0167] 23. The pharmaceutical composition of embodiment 20, 21, or 22, wherein the pH adjuster is NaOH.

[0168] twenty four. (a) Below: (ai) radionuclide 177 Lutetium (Lu-177) and (aii) Formula (I): [ka] with NeoB and the complex formed by; (b) gentisic acid or its salts and ascorbic acid or its salts; (c) macrogol 15 hydroxystearate and; (d) acetate buffer; (e) Water for injection; (f) NaOH and; (g) DTPA, and a pharmaceutical composition comprising the same.

[0169] 25. The pharmaceutical composition according to any one of embodiments 1 to 24, wherein the pharmaceutical composition is an aqueous solution.

[0170] 26. The pharmaceutical composition according to any one of embodiments 1 to 25, wherein the pharmaceutical composition is an infusion solution.

[0171] 27. A pharmaceutical composition according to any one of embodiments 1 to 26 for use in the treatment or prevention of cancer, typically GRPR-positive cancer.

[0172] 28. The pharmaceutical composition of any one of embodiments 1 to 27, wherein the solution is produced in a commercial scale manufacturing, in particular in a batch size of at least 0.5 Ci.

[0173] 29. A pharmaceutical composition according to any one of embodiments 1 to 28, which is for commercial use.

[0174] 30. A process for producing the pharmaceutical composition as defined above, comprising: (1) A complex of a radionuclide and a GRP receptor peptide antagonist binding moiety linked to a chelator is (1.1) preparing an aqueous solution containing only a radionuclide and one stabilizer against radiolysis, which is gentisic acid or its salts; (1.2) preparing an aqueous solution comprising a GRP receptor peptide antagonist binding moiety linked to a chelator and, optionally, a surfactant; and (1.3) mixing the solutions obtained in steps (1.1) and (1.2), heating the resulting mixture, and optionally filtering the resulting complex; and forming a process step by; (2) The complex solution obtained in step (1) (2.1) preparing a dilute aqueous solution optionally containing only one stabilizer against radiolysis, which is ascorbic acid; and (2.2.) Mixing the complex solution obtained in step (1) with the diluted solution obtained in step (2.1) to obtain a final solution; and a process step of diluting by

[0175] 31. The solution from step (1.1) is 177 31. The process of embodiment 30, comprising LuCl3 and HCl.

[0176] 32. The process according to any one of embodiments 30 to 31, wherein the solution prepared in step (1.1) contains only one stabilizer, which is gentisic acid or a salt thereof, at a concentration of at least 1000 μg / mL, for example, from 1000 μg / mL to 1500 μg / mL.

[0177] 33. The process according to any one of embodiments 30 to 32, wherein the solution of step (1.1) further comprises a buffer, preferably an acetate buffer.

[0178] 34. The process of any one of embodiments 30-33, wherein the GRP receptor peptide antagonist binding moiety linked to a chelator in the solution of step (1.2) is NeoB of formula (I). [ka]

[0179] 35. The process of any one of embodiments 30-34, wherein the solution of step (1.2) further comprises a surfactant that is macrogol 15 hydroxystearate.

[0180] 36. The process according to any one of embodiments 30 to 35, wherein the solution prepared in step (2.1) contains only one stabilizer, which is ascorbic acid or a salt thereof, at a concentration of at least 10000 μg / mL, preferably at least 12000 μg / mL, preferably at least 15000 μg / mL, for example 12000 to 18000 μg / mL.

[0181] 37. The process according to any one of embodiments 30 to 36, wherein the solution of step (1.2) further comprises macrogol 15 hydroxystearate at a concentration comprised between 5 μg / mL and 5000 μg / mL, preferably between 25 μg / mL and 2000 μg / mL, more preferably between 50 μg / mL and 1000 μg / mL, and even more preferably at a concentration of 100 μg / L.

[0182] 38. The process of any one of embodiments 30 to 37, wherein in step (1.3), the resulting mixture is heated to a temperature of 70 to 99°C, preferably 90 to 98°C, for 1 to 59 minutes, preferably 2 to 15 minutes.

[0183] 39. The process according to any one of embodiments 30 to 38, wherein the complex obtained at the end of step (1.3) is further filtered at 0.20 μm.

[0184] 40. The process of any one of embodiments 30-39, wherein the solution of step (2.1) further comprises a sequestering agent that is diethylenetriaminepentaacetic acid (DTPA) or a salt thereof.

[0185] 41. The process of any one of embodiments 30-40, wherein the solution of step (2.1) further comprises a pH adjuster that is NaOH.

[0186] 42. The process of any one of embodiments 30-41, wherein the solution of step (2.1) further comprises water for injection.

[0187] 43. The process according to any one of embodiments 30 to 42, wherein (3) a process step of sterile filtering the solution obtained in step (2); (4) aseptically dispensing the filtered solution obtained in step (3) into unit-dose containers, wherein the radionuclide is present at a concentration that results in a volumetric activity of at least 370 MBq / mL (at EOP) ± 37 MBq / mL (± 10%).

[0188] 44. The process of any one of embodiments 30-43, wherein the dose unit containers of step (4) are stoppered vials sealed within a lead container.

[0189] 45. An aqueous pharmaceutical solution obtained by a process defined by any one of embodiments 30 to 44. [Example]

[0190] The present disclosure will now be described in more detail, particularly with reference to examples, which are not intended to limit the present invention.

[0191] material: 177 LuCl3 can be obtained from commercial sources, such as IDB Holland BV. All other ingredients of the drug are commercially available from a variety of sources.

[0192] Methods for preparing pharmaceutical compositions The preparation of Lu-NeoB was carried out automatically using a MiniAIO synthesizer. The synthesis procedure was formulated as follows: 1. 177 Transfer LuCl3 into the reactor; 2. Transfer the reaction buffer into the reactor. The reaction buffer is composed of sodium acetate buffer and gentisic acid. The acetate buffer can maintain the pH of the labeling at 4-5, and the gentisic acid protects the peptide from radiolysis during the labeling step; 3. Add NeoB solution containing Kolliphor HS 15 to the reactor; 4. Heat at 95°C for 5 minutes; At the end of labeling, a diluent solution is added to obtain a volumetric radioactivity of 10 mCi / mL, which consists of ascorbic acid (antioxidant), DTPA (sequestering agent), NaOH (pH adjuster), and water for injection.

[0193] Example 1: Effect of formulation on radiochemical purity of pharmaceuticals Contains the same amount of antioxidants as present in Lutathera formulations 177 Stability of Lu-labeled product over 72 hours. Specifically, the following conditions are reproduced: Gentisic acid 630 μg / mL is added before the labeling step; Kolliphor HS 15: 1 mg is added before the labeling step; ○ Peptide:Lu ratio ≥ 1.5; o Addition of 2795 μg / mL of ascorbic acid at the end of the reaction during the compounding step; ○Final volume radioactivity 10mCi / mL; ○Final pH 4~6; Reaction buffer: acetic acid / acetate buffer;

[0194] Radiolabeling tests are performed both manually and automatically using the MiniAIO synthesizer. The synthesis procedure is formulated as follows: 1. 177 Transfer LuCl3 into the reactor; 2. Transfer the reaction buffer into the reactor. The reaction buffer is composed of sodium acetate buffer and gentisic acid. The acetate buffer can maintain the pH of the labeling at 4-5, and the gentisic acid protects the peptide from radiolysis during the labeling step; 3. Add NeoB1 solution containing Kolliphor HS 15 to the reactor; 4. Heat at 95°C for 5 minutes; At the end of labeling, a diluent solution is added to obtain a volumetric radioactivity of 10 mCi / mL, which consists of ascorbic acid (antioxidant), DTPA (sequestering agent), NaOH (pH adjuster), and water for injection.

[0195] [Table 1]

[0196] As the results shown in Table 1 demonstrate, a product formulated under the same conditions as Lutathera gradually deteriorates over time, ultimately resulting in a radiochemical purity below specification at the end of the target shelf life (72 hours). 177 The development of LuNeoB has focused on identifying the appropriate amounts of gentisic acid and ascorbic acid that can exert the desired protective function without interfering with the labeling step.

[0197] Example 2: Identification of suitable formulations to improve the radiochemical purity of pharmaceuticals Antioxidants / free radical scavengers such as ascorbic acid and gentisic acid are commonly used in the preparation of radiopharmaceuticals to protect the labeled molecule from radiolysis.

[0198] Therefore, to identify the appropriate formulation for improving radiochemical purity, various formulations are tested by increasing the amount of gentisic acid or ascorbic acid while keeping all other conditions constant, including the amount of other antioxidants.

[0199] ·Change in the proportion of gentisic acid First, we investigate the effect of varying the proportion of gentisic acid. To do so, we test increasing amounts of gentisic acid while keeping all other conditions constant, including the amount of ascorbic acid.

[0200] Different concentrations of gentisic acid are added up to 1000 μg / mL as described in the table below and tested under the following conditions: Gentisic acid is added before the labeling step; Kolliphor HS 15: 1 mg is added before labeling; Add 2700 μg / mL of ascorbic acid at the end of labeling during the compounding step; ○Final volume radioactivity 10mCi / mL; ○ Peptide:Lu ratio ≥ 1.5; ○Final pH 4~6; Reaction buffer: acetic acid / acetate buffer;

[0201] [Table 2]

[0202] The results shown in Table 2 indicate that when the concentration of ascorbic acid was set at 2700 μg / mL and the maximum concentration of gentisic acid was set at 1000 μg / mL, the radiochemical purity did not meet the specification at the end of the shelf life.

[0203] Therefore, altering the proportion of gentisic acid does not affect the radiochemical purity.

[0204] -Change in the proportion of ascorbic acid We then investigate the effect of different amounts of ascorbic acid on the stability of the final product. As shown in the table below, different concentrations of ascorbic acid are added up to 15000 μg / mL and tested under the following conditions: Gentisic acid 1000 μg / mL added before the labeling step; Kolliphor HS 15: 1 mg is added before labeling; Adding ascorbic acid at the end of labeling during the compounding step; ○Final volume radioactivity 10mCi / mL; ○ Peptide:Lu ratio ≥ 1.5; ○Final pH 4~6; Reaction buffer: acetic acid / acetate buffer;

[0205] [Table 3]

[0206] As shown by the results in Table 3, the radiochemical purity of the drug product is greater than 95% for up to 72 hours in formulations containing at least 1000 μg / mL gentisic acid and 10000 μg / mL ascorbic acid. Tests performed with increasing ascorbic acid concentrations show a clear improvement in the stability of the product at 15000 μg / mL (RCP% >98% at the end of the shelf life).

[0207] Evaluation of the antioxidant properties of ascorbic acid by removing gentisic acid from liquid formulations Based on the results shown in Table 3, the minimum amount of ascorbic acid for these studies was set at 15,000 μg / mL. As noted in Table 4, gentisic acid was not part of the formulation and only ascorbic acid was present as an antioxidant. The following conditions apply: Kolliphor HS 15: 1 mg is added before labeling; Add 15000 μg / mL of ascorbic acid at the end of labeling during the compounding step; ○Final volume radioactivity 10mCi / mL; ○ Peptide:Lu ratio ≥ 1.5; ○Final pH 4~6; Reaction buffer: acetic acid / acetate buffer;

[0208] [Table 4]

[0209] As can be seen, in the absence of gentisic acid, the radiochemical purity of the drug product after 72 hours is below 95%. Therefore, based on the results of this experiment, gentisic acid and ascorbic acid have complementary positive effects on drug product stability.

[0210] In conclusion, the best results regarding the radiochemical stability of the product were obtained with a formulation containing both gentisic acid and ascorbic acid at concentrations of 1000 μg / mL and 15000 μg / mL, respectively.

[0211] Example 3: Final formulation test at 200 mCi The following studies are designed to confirm the formulations identified through previous development studies (Examples 1-3) at a radioactivity level of 200 mCi. Based on the previous results, the amounts of gentisic acid and ascorbic acid are set to 1000 μg / mL and 15000 μg / mL, respectively. The synthesis is carried out under the following conditions: 1000 μg / mL of gentisic acid is added to the reactor from the beginning; Add Kolliphor HS 15 to the aqueous peptide solution to a final concentration of 100 μg / mL; Add 15000 μg / mL of ascorbic acid at the end of labeling during the compounding step; Final volume radioactivity after compounding: 10 mCi / mL; ○ Peptide:Lu ratio ≥ 1.5; ○Final pH 4~6; Reaction buffer: acetic acid / acetate buffer;

[0212] Radiolabeling is automated using a MiniAIO synthesizer. The synthesis procedure is formulated as follows: 1. 177 Transfer LuCl3 into the reactor; 2. Transfer the reaction buffer into the reactor. The reaction buffer is composed of sodium acetate buffer and gentisic acid. The acetate buffer can maintain the pH of the labeling at 4-5, and the gentisic acid protects the peptide from radiolysis during the labeling step; 3. Add NeoB1 solution containing Kolliphor HS 15 to the reactor; 4. Heat at 95°C for 5 minutes; 5. At the end of labeling, a dilution solution is added to obtain a volumetric radioactivity of 10 mCi / mL. The dilution solution is composed of ascorbic acid (antioxidant), DTPA (sequestering agent), NaOH (pH adjuster), and saline.

[0213] As the results shown in Table 7 show, using 1000 μg / mL gentisic acid and 15000 μg / mL ascorbic acid, for a level of radioactivity of 200 mCi: 177 The radiochemical purity of LuNeoB is consistently well above 95% for up to 72 hours.

[0214] [Table 5]

[0215] Example 4: Final formulation test on 0.5 Ci batches Based on the results obtained during R&D lab-scale product development, the following composition was selected for the initial scale-up batch production: Gentisic acid 1000 μg / mL; Ascorbic acid 15000μg / mL; Kolliphor HS 15 100 μg / mL; ○Volumetric radioactivity 10 mCi / mL; ○Final pH 4.0~6.0; Reaction buffer: acetic acid / acetate buffer;

[0216] To transition from R&D formulation to pharmaceutical quality drug product, scale-up batches were produced using the reaction buffer (product code F193) and formulation buffer (product code F191) produced by Gipharma and used in the production of Lutathera. 177 The stability of the LuNeoB final product was evaluated in three different sample volumes (4 mL, 6 mL, and 25 mL) stored at 25±2°C for up to 72 hours.

[0217] To industrialize the production of the drug substance, the scale-up study also aimed to optimize the manufacturing process carried out with the assistance of an automated synthesis module. 177 A drug substance (mother liquor) containing the Lu-labeled molecule is prepared.

[0218] An automated synthesis process was developed to produce the radioactive drug substance as a sterile, concentrated aqueous mother liquor. The drug substance synthesis steps were configured in a MiniAIO (TRASIS) synthesizer module, a stand-alone, closed-system synthesis module automated and remotely controlled by GMP-compliant software that monitored and recorded process parameters.

[0219] The Mini AIO radiosynthesis module is widely used in the radiopharmaceutical industry for the production of PET radiopharmaceuticals. The module incorporates a disposable flow path, which is preferred over fixed flow path devices because it ensures a sterile, pyrogen-free flow path and eliminates the possibility of cross-contamination between batches. The synthesis module is placed in a lead-shielded hot cell supplied with Grade C HEPA-filtered air. Inside the clean Grade C laboratory is an isolator.

[0220] Table 8 lists the target formulation properties selected for the production of a 0.5 Ci batch size.

[0221] [Table 6]

[0222] ○ 177 Theoretical activity of LuCl3 = 0.5 Ci; ○When synthesis begins 177 Specific activity of LuCl3 = 9.7 Ci / mg; ○Net amount of NeoB1 = 0.600 mg; ○ Molar ratio (NeoB1:Lu) = 1.300; Synthesis using MiniAIO module (Trasis); ○Labeling time: 5 minutes; ○Labeling temperature: 95℃;

[0223] 177 Some relevant IPC results obtained during the manufacture of the LuNeoB1.0.5 Ci batch size are listed in Table 9.

[0224] [Table 7]

[0225] To assess the effect of oxidative degradation, three sample volumes were dispensed at the end of production: · 25mL (vial-1); · 4mL (Vial-2, Vial-3, Vial-4); · 6mL (vial-5);

[0226] In sample vial-4, an additional amount of gentisic acid was added during the compounding step to a final concentration of approximately 1.312 mg / mL, which ultimately further reduced radiolysis. Sample vial-3 was maintained under agitation throughout the stability study.

[0227] All samples were stored at 25±2° C. The various samples dispensed are shown in Table 10.

[0228] [Table 8]

[0229] Table 11 shows 177 1 is a summary of the radiochemical purity results obtained for a scaled-up batch of LuNeoB1 0.5 Ci. As can be seen, the radiochemical purity of the product at time t0 meets the target specification of >97.00%.

[0230] Stability testing performed on Vial-1 (25 mL sample) shows very promising results even after 72 hours, while Vial-2 (4 mL sample) and Vial-3 (4 mL sample stirred) show radiochemical purity below 95.00% at the end of the target shelf life. These preliminary results appear to indicate a detrimental effect of O on the stability of the final product.

[0231] Adding additional amounts of gentisic acid (sample vial-4) improved the stability of the final product, but did not successfully meet the target shelf life. Finally, Vial-5 (6 mL sample) shows improvement in terms of stability results compared to the 4 mL sample volume, despite the radiochemical purity at 72 hours not meeting specifications.

[0232] [Table 9]

[0233] Final target formulation and composition details The final amounts of ascorbic acid and gentisic acid were defined based on data collected during the development activities. In particular, gentisic acid at a concentration of 1312 ppm, together with ascorbic acid at a concentration of 15000 ppm, showed excellent antioxidant properties that allowed the target shelf life to be achieved.

[0234] Based on all development tests performed, radioactivity levels up to 500 mCi 177 The formulation chosen for the production of LuNeoB is as follows:

[0235] Table 10

Claims

1. (a) a complex comprising: (ai) radioactive nuclide １７７ Lu, and (aii) Compounds of formula (I) 【Chemistry 1】 the complex formed by (b) at least two stabilizers against radiolysis comprising gentisic acid and ascorbic acid, wherein said gentisic acid and said ascorbic acid are present in a weight ratio of 1:16 to 1:10; (c) optionally a surfactant; A pharmaceutical composition comprising: the pharmaceutical composition has a radiochemical purity of greater than 95% for at least 72 hours; the gentisic acid or salt thereof is present at a concentration of at least 1000 μg / mL; The pharmaceutical composition, wherein the ascorbic acid or a salt thereof is present in a concentration of at least 10,000 μg / mL.

2. 2. The pharmaceutical composition of claim 1, wherein the radionuclide is present at a concentration that results in a volumetric activity of at least 370 MBq / mL (end of process) ± 37 MBq / mL (± 10%).

3. The pharmaceutical composition described in claim 1, wherein the ratio between the gentisic acid and the ascorbic acid is 1:15 to 1:

10.

4. The pharmaceutical composition described in claim 1, wherein the ratio between the gentisic acid and the ascorbic acid is 1:12 to 1:

11.

5. 5. The pharmaceutical composition according to any one of claims 1 to 4, wherein the gentisic acid or a salt thereof is present in a concentration of 1000 μg / mL to 1500 μg / mL.

6. 6. The pharmaceutical composition of any one of claims 1 to 5, wherein the ascorbic acid or salt thereof is present in a concentration of at least 12000 μg / mL, or at least 15000 μg / mL.

7. 6. The pharmaceutical composition according to any one of claims 1 to 5, wherein the ascorbic acid or a salt thereof is present at a concentration of 12,000 to 18,000 μg / mL.

8. A pharmaceutical composition described in any one of claims 1 to 7, wherein the ascorbic acid or its salt is present at a concentration of at least 12,000 μg / mL.

9. 8. The pharmaceutical composition according to claim 1, wherein the gentisic acid or a salt thereof is present at a concentration of 1000 μg / mL to 1500 μg / mL, and the ascorbic acid or a salt thereof is present at a concentration of 12000 to 18000 μg / mL.

10. 10. The pharmaceutical composition of any one of claims 1 to 9, wherein said pharmaceutical composition has a radiochemical purity of greater than 98% for at least 72 hours.

11. (a) a complex comprising: (ai) radioactive nuclide １７７ Lutetium (Lu-177), and (aii) NeoB of formula (I) 【Chemistry 2】 the complex formed by (b) gentisic acid or a salt thereof and ascorbic acid or a salt thereof; (c) optionally, macrogol 15 hydroxystearate; (d) optionally, at least one other pharmaceutically acceptable excipient; A pharmaceutical composition comprising: the gentisic acid or salt thereof is present at a concentration of at least 1000 μg / mL; the ascorbic acid or salt thereof is present at a concentration of at least 10,000 μg / mL, at least 12,000 μg / mL, or at least 15,000 μg / mL; the pharmaceutical composition has a radiochemical purity of greater than 95% for at least 72 hours; The pharmaceutical composition, wherein the gentisic acid and the ascorbic acid are present in a weight ratio of 1:16 to 1:

10.

12. 12. The pharmaceutical composition according to claim 11, wherein the at least one other pharmaceutically acceptable excipient is selected from a buffer, and / or a solvent, and / or a pH adjusting agent.

13. (a) a complex comprising: (ai) radioactive nuclide １７７ Lutetium (Lu-177), and (aii) NeoB of formula (I) 【Transformation 3】 the complex formed by (b) gentisic acid or a salt thereof and ascorbic acid or a salt thereof; (c) macrogol 15 hydroxystearate; (d) acetate buffer; (e) water for injection; (f) with NaOH; (g) DTPA and A pharmaceutical composition comprising: the gentisic acid and the ascorbic acid are present in a weight ratio of 1:16 to 1:10; the pharmaceutical composition has a radiochemical purity of greater than 95% for at least 72 hours; the gentisic acid or salt thereof is present at a concentration of at least 1000 μg / mL; The pharmaceutical composition, wherein the ascorbic acid or a salt thereof is present in a concentration of at least 10,000 μg / mL.

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