Radiolabeling and Formulation for Scale-Up of 64Cu-DOTATATE

Abstract

The present disclosure relates to methods for creating robust procedures capable of supplying commercial quantities of radioactive diagnostic agents adapted for use in positron emission tomography (PET) for the localization of somatostatin receptor-positive neuroendocrine tumors (NETs) in adult patients. Provided herein are: 64 Cu-DOTATATE preparation, final formulation parameters and 64 Design of Experiments (DOE) to monitor the effect on Cu-DOTATATE stability, 500 mCi to 2000 mCi 64 Scaled-up experiments to prepare Cu-DOTATATE and its final formulation 64 The stability of Cu-DOTATATE, optimization of the amount of DOTATATE used relative to the total activity used for radiolabeling, and 64 Cu-DOTATATE purity 64 Effect of Cu on the specific activity of copper chloride solution.

Description

[Technical Field]

[0001] Related Projects This application claims priority to U.S. Provisional Application No. 63 / 074,451, filed September 3, 2020, which is incorporated by reference herein in its entirety to the maximum extent permitted by law.

[0002] Technical Field The present disclosure provides bioconjugate compounds containing positron-emitting radionuclides. 64 The present invention relates to compositions and methods for the radiolabeling and purification of Cu-DOTATATE. [Background technology]

[0003] background The most important known imaging techniques in medical diagnosis are positron emission tomography (PET), computed tomography (CT), magnetic resonance imaging (MRI), single-photon computed tomography (SPECT), and ultrasound (US). Although today's imaging technologies are well developed, most rely on non-specific macroscopic, physical, physiological, or metabolic changes to distinguish normal from pathological tissue.

[0004] Targeting molecular imaging (MI) has the potential to reach new dimensions in medical diagnostics. The term "targeting" refers to the selective and highly specific binding of natural or synthetic ligands (binders) to molecules of interest (molecular targets) in vitro or in vivo.

[0005] MI is a rapidly emerging field of biomedical research that can be defined as the visual representation, characterization, and quantification of biological processes at the cellular and subcellular level within intact, living organisms. It is a novel interdisciplinary field, and the images generated do not identify the molecular events that cause disease, but rather reflect the cellular and molecular pathways and in vivo mechanisms of disease as they exist in the context of a physiologically authentic environment.

[0006] Several different contrast-enhancing agents are known today. They can be used in functional imaging, primarily developed for PET and SPECT. The application of radiolabeled biologically active peptides for diagnostic imaging is gaining importance in nuclear medicine. Biologically active molecules that selectively interact with specific cell types are useful for delivering radioactivity to target tissues. For example, radiolabeled peptides have significant potential for delivering radionuclides to tumors, infarcts, and infected tissues for diagnostic imaging and radiotherapy.

[0007] DOTA (1,4,7,10-tetrakis(carboxymethyl)-1,4,7,10-tetraazacyclododecane) and its derivatives constitute an important class of chelators for biomedical applications because they accommodate a variety of divalent and trivalent metal ions with great stability. One of its derivatives, DOTATATE, [(4,7,10-tricarboxymethyl-1,4,7,10-tetrazacyclododec-1-yl)acetyl]-(D)-phenylalanyl-(L)-cysteinyl-(L)-tyrosyl-(D)-tryptophanyl-(L)-lysyl-(L)-threoninyl-(L)-cysteinyl-(L)-threonine-cyclic(2-7)disulfide, can be used as a targeting agent. The chemical structure of DOTATATE is shown below. [ka]

[0008] An emerging area is the use of bioactive peptides conjugated with chelators for labeling with radiometals in various areas of diagnostic and therapeutic nuclear oncology. There have been several reports in recent years on targeted radiotherapy with radiolabeled somatostatin analogues. 68 Ga-DOTATATE (Dedden SA, et al.; J Nucl Med; 2016 vol. 57 no. 6 872-878), 68Ga-DOTATOC (Nicolas, GP, et al.; J Nucl Med; 2018 vol. 59 no. 6 915-921), 68 Ga-DOTANOC (Amdrosini V., et al.; J Nucl Med; 2010 vol. 51 no. 5 669-673) is a known PET tracer used to visualize NETs. 177 Lu-DOTATATE is used in radionuclide therapy (Strosberg, J. et al.; N Engl J Med 2017; 376:125-135). However, there is a need for additional peptide-based compounds that have utility in diagnostic imaging techniques such as PET.

[0009] Copper-64( 64 Cu) is a positron-emitting radionuclide well suited for use as a diagnostic agent for positron emission tomography (PET). Its half-life of 12.7 hours is long enough to allow for post-production processing, labeling, and transport, and its mean positron energy of 0.28 MeV provides high-resolution images. Importantly, 64 Ni(p,n) 64 The broad cross-section of Cu reactions allows for the production of commercial quantities. The PET radioisotope copper-64 (Cu-64) has been radiolabeled to the chelate-peptide conjugate DOTATATE for diagnostic imaging of human neuroendocrine tumors.

[0010] Although the complete chemical structure of the Cu-DOTATATE complex has not been determined by X-ray crystallography, the Cu-DOTA complex has been structurally determined by X-ray crystallography. In crystalline form, the Cu-DOTA complex has been shown to be hexa-coordinated, utilizing four amino nitrogen atoms and two carboxylate oxygen atoms, as described below. [ka]

[0011] Two of the carboxylic acid groups remain free and are not coordinated to the copper metal ion, so attachment of the peptide through one of the carboxylic acids to form a linking amide bond is not expected to alter the coordination of copper to the DOTATATE peptide.

[0012] 64 Cu-DOTATATE binds to somatostatin receptors with highest affinity to the subtype 2 receptor (SSTR2). It binds to cells expressing somatostatin receptors, including malignant neuroendocrine cells that overexpress SSTR2 receptors. 64 Cu is a positron (β) carrier with an emission yield that allows for positron emission tomography (PET) imaging. + ) is an emitting radionuclide. 64 When the imaging capabilities of Cu were combined with the receptor targeting capabilities of DOTATATE, the results showed that it was possible to image somatostatin receptor-expressing neuroendocrine tumors (NETs). 64 Cu-DOTATATE, a radiopharmaceutical agent. Today, 64 Cu-DOTATATE is prepared for on-site use with low total radioactivity for a very limited number of patients. Therefore, high purity 64 To provide an improved process for making Cu-DOTATATE, 64 There remains an unmet need to scale up radiolabeled production of Cu-DOTATATE while maintaining sufficient stability for transport of the drug product to patients within the hospital. [Prior art documents] [Non-patent literature]

[0013] [Non-Patent Document 1] Dedden SA, et al.; J Nucl Med; 2016 vol. 57 no. 6 872-878 [Non-patent document 2] Nicolas, GP, et al.; J Nucl Med; 2018 vol. 59 no. 6 915-921 [Non-patent document 3] Amdrosini V., et al.; J Nucl Med; 2010 vol. 51 no. 5 669-673 [Non-patent document 4] Strosberg, J. et al.; N Engl J Med 2017; 376:125-135 Summary of the Invention [Means for solving the problem]

[0014] overview The present disclosure meets the above needs and provides a copper-labeled drug product, 64 The present invention relates to a method that provides a useful process capable of supplying commercial quantities of Cu-DOTATATE.

[0015] The purpose of the present invention is to demonstrate and confirm the discovery that labeling copper at lower temperatures (≦30° C.) has the advantage of improving the purity of drug products, since many other common metal impurities actually label significantly slower than copper to chelates such as DOTATATE. 64 Cu, 67 Previous studies on radiolabeling of Cu (Cu) have typically been performed at elevated temperatures, such as 40°C to 95°C. High temperatures were used to speed up the labeling process and ensure maximum radiolabeling of copper to the chelate. Some literature reviews indicate that labeling at room temperature can achieve sufficient labeling. The present disclosure teaches that the faster labeling kinetics of copper can be used to obtain a purer product compared to the slower labeling kinetics of other metals.

[0016] Provided herein are: 64 Cu-DOTATATE preparation, final formulation parameters and 64Design of Experiments (DOE) to monitor the effect on Cu-DOTATATE stability, 500 mCi to 2000 mCi 64 Scaled-up experiments to prepare Cu-DOTATATE and its final formulation 64 The stability of Cu-DOTATATE, optimization of the amount of DOTATATE used relative to the total activity used for radiolabeling, and 64 Cu-DOTATATE purity 64 Effect of Cu on the specific activity of copper chloride solution.

[0017] For example, provided herein is a method for radiolabeling DOTATATE, comprising reacting copper-64 with a buffered solution containing DOTATATE, wherein the reaction occurs in less than 15 minutes at a temperature less than or equal to 30°C, and wherein the molar ratio of DOTATATE to copper-64 in the reaction solution is from about 110:1 to about 90:1.

[0018] Further herein, 64 A method is provided for preparing a drug product comprising Cu-DOTATATE, wherein the drug product is prepared by (i) radiolabeling DOTATATE with copper-64 at a concentration of about 0.6 μg / mL (μg of DOTATATE per mCi of copper-64), and wherein the radionuclide purity of the copper-64 in the drug product is about 99%.

[0019] In this specification, 64 1. A drug product for use in positron emission tomography, comprising Cu-DOTATATE, 64 Cu-DOTATATE, 148MBq 64 Also provided is a drug product stored in a single-dose vial containing Cu-DOTATATE, the drug product having a radioactivity concentration of about 5-15 mCi / mL and a radiochemical purity of ≥96% after dilution.

[0020] The foregoing features of the embodiments will be more readily understood by reference to the following detailed description taken in conjunction with the accompanying drawings, in which: [Brief explanation of the drawings]

[0021] [Figure 1A] FIG. 1(A) presents a schematic radiolabeling and formulation scheme. [Figure 1B] FIG. 1(B) presents a schematic radiolabeling and formulation scheme for the present invention.

[0022] [Figure 2] FIG. 2 presents representative HPLC chromatograms of standard solutions of gentisic acid and DOTATATE.

[0023] [Figure 3] Figure 3 presents a representative HPLC chromatogram of the crude Cu-DOTATATE reaction mixture after mixing equimolar amounts of DOTATATE and Cu at room temperature for 5 minutes.

[0024] [Figure 4] Figure 4 presents the recovery of DOTATATE in the fraction loading solution (12 mL total) and the final 50% EtOH eluate at a flow rate of 12 mL / min.

[0025] [Figure 5] Figure 5 presents the recovery of DOTATATE in the fraction loading solution (18 mL total) and the final 50% EtOH eluate at a flow rate of 18 mL / min. DETAILED DESCRIPTION OF THE INVENTION

[0026] Detailed Description Various aspects and embodiments will now be fully described herein. However, these aspects and embodiments may be embodied in many different forms and should not be construed as limiting; rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the subject matter to those skilled in the art. All publications, patents, and patent applications cited herein, whether supra or infra, are hereby incorporated by reference in their entirety. A.Definition

[0027] Unless otherwise defined, all terms and phrases used herein include the meaning acquired by the term in the art unless expressly stated to the contrary or made clear by the context in which the term or phrase is used. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, particular methods and materials are now described.

[0028] Unless otherwise stated, the use of individual numerical values ​​is described as an approximation, as if the value were preceded by the word "about" or "approximately." Similarly, various ranges of numerical values ​​specified in this application are described as approximations, as if both the minimum and maximum values ​​within the stated range were preceded by the word "about" or "approximately," unless otherwise expressly stated. In this manner, variations above and below the stated range can be used to achieve substantially the same results as values ​​within the range. As used herein, the terms "about" and "approximately," when referring to numerical values, shall have their plain and ordinary meaning to one of ordinary skill in the art to which the disclosed subject matter most closely pertains or to which the range or element in question pertains. The amount of spread from an exact numerical boundary depends on many factors. For example, some of the factors that may be considered include the importance of the element and / or the impact that a given amount of variation will have on the performance of the claimed subject matter, as well as other considerations known to those of ordinary skill in the art. As used herein, the use of different amounts of significant digits for different numerical values ​​is not intended to limit how the use of the word "about" or "approximately" acts to spread a particular numerical value or range. Thus, as a general matter, "about" or "approximately" broadens the numerical value. Also, the disclosure of ranges is intended as a continuous range that includes every value between the minimum and maximum values, as well as the broadening of the range obtained by use of the term "about" or "approximately." Consequently, the recitation of ranges of values ​​herein is merely intended to serve as a shorthand method of referring individually to each separate value within the range, and each separate value is incorporated herein as if it were individually recited herein.

[0029] "drug products" or " 64 The term "Cu-DOTATATE injection" is used interchangeably herein and refers to the compound in its final formulation used as a radioactive diagnostic agent. 64 This refers to Cu-DOTATATE.

[0030] "Optionally" or "optionally" means that the subsequently described element, component, or circumstance may or may not occur, and thus the description includes instances where the element, component, or circumstance occurs and instances where it does not occur.

[0031] The terms "subject" and "patient" are used interchangeably herein and refer to a human or other mammal. B. Introduction

[0032] The present disclosure provides: 64 Improved radiolabeling and formulation for scale-up of Cu-DOTATATE preparation; Figure 1(A).

[0033] High-resolution imaging modalities of positron emission tomography (PET) can be used in oncology to help clinicians gain a better understanding of a patient's disease state, monitor the effectiveness of treatment, and provide more effective, personalized care. One such PET agent targets and images neuroendocrine tumors (NETs) that overexpress somatostatin receptor subtype 2 (SSTR2). 64 The imaging capability is a positron-emitting radionuclide, Cu-DOTATATE, which can help identify patients who may benefit from receptor-targeted treatment. 64 Cu (t 1 / 2 =12.7 hours, β + avg = 0.28 MeV, I = 17.6% [sometimes reported as branching ratio (BR), representing intensity (I)], which can be imaged using PET, but the targeting moiety of the molecule is a modified version of octreotate (DOTA- D -Phe-Cys-Tyr- D -Trp-Lys-Thr-Cys-Thr, disulfide cyclized Cys2-Cys7), a cyclic peptide that mimics the natural SSTR2-ligand somatostatin. These two functions are simultaneously 64 They are bound together by DOTA, a bifunctional chelator that captures Cu while remaining attached to the N-terminus of the peptide (forming DOTATATE).64 The structure of Cu-DOTATATE (copper Cu64 DOTATATE) is shown below. [ka]

[0034] radioactive isotope 64 Radiolabeling of DOTATATE with Cu was initially performed several decades ago. 64 Cu-DOTATATE was prepared at low total activity for very limited use and a limited number of patients, and for on-site use only. Recently, radiolabeling has been improved to higher activity for scale-up to commercial production. The final purified product was made at a much higher initial total activity level, and radiolysis was prevented by improved formulation and purification methods.

[0035] Scaling up the radiolabeling and formulation of large quantities of drugs was necessary to enable drug distribution across the country. Scaling up brought new problems and challenges, as well as new discoveries and solutions, to achieve large batches of drugs. This disclosure (i) describes improvements and modifications to previous studies and results, and (ii) teaches studies that have been performed and teaches scale-up to large, highly active batches.

[0036] The general radiolabeling and formulation scheme that has been used is shown in Figure 1(B).

[0037] Specifically, the present disclosure provides: 64 Cu-DOTATATE can be radiolabeled and purified for injectable drug products 64 This paper teaches a significant scale-up of the total radioactivity of Cu.

[0038] In previous studies, 64 The radioactivity (mCi) of Cu was low to moderate. This study scaled up the radiolabeling to a total radioactivity of >5,400 mCi. The challenge was due to radiolysis and the presence of DOTATATE. 64The goal is to achieve radiolabeling without decomposition due to competition with other metals other than Cu. The radiolabeled product must then be rapidly purified to maintain the required high radiochemical purity (RCP) and immediately diluted in a stabilizing solution to prevent degradation from radiolysis.

[0039] Purified in its final formulation (45 mg / mL in sodium ascorbate, 5% ethanol) 64 The stability of Cu-DOTATATE was evaluated for up to 48 hours after labeling, and the release of ATP from the complex was confirmed. 64 It shows that there is no significant decomposition or loss of Cu.

[0040] 64 To prepare Cu-DOTATATE, 64 CuCl is reacted with DOTATATE in a sodium acetate buffer containing gentisic acid at a ratio of 2 μg DOTATATE / mCi. The reaction mixture is incubated and then purified in a sodium ascorbate (NaOAsc) buffer. 64 The Cu-DOTATE solution was sterile filtered to form the final formulation. The development efforts disclosed herein aim to improve the production design space and to develop a Cu-DOTATE solution with a yield of ≥ 2 Ci. 64 This paper focuses on scaling up the radiolabeling reaction to prepare Cu-DOTATATE. Radiolabeling was demonstrated even at 15°C for 5 minutes. The purified product was purified to yield up to 10,000 mCi. 64 This was achieved with Cu-DOTATATE. Purified product was achieved in 50% ethanol in water (previous literature showed only pure ethanol). In fact, the use of 50% ethanol in water improved the yield of purified product compared to the use of 100% ethanol.

[0041] The purified drug product (2 mL) was immediately diluted to a larger volume (>20 mL; but typically >100 mL for 2000-10,000 mCi products) to prevent degradation (radiolysis) and maintain the required RCP >95%. Previous literature has diluted the purified product to <20 mL.

[0042] The final drug product was stabilized for up to 48 hours after purification using 28-122 mg / mL sodium ascorbate with 1-5% ethanol at an RCP >95%. Previous RCP stabilization for 48 hours was only achieved with 45 mg / mL sodium ascorbate / 5% ethanol.

[0043] The initial labeling (radiolabeling step) can be achieved in the presence of sodium ascorbate.

[0044] Surprisingly, it was found that chelation of copper by DOTATATE occurs more rapidly than other metals when labeled with DOTATATE at lower temperatures, i.e., ≦30° C. This phenomenon can be used to reduce the amount of metallic impurities present in the final drug product. C. 64 Preparation of Cu-DOTATATE bulk solution i. Ligand

[0045] In one embodiment, the ligand is DOTATATE; 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA); 3,6,9,15-tetraazabicyclo[9.3.1]pentadeca-1(15),11,13-triene-3,6,9-triacetic acid (PCTA); 1,4,7-triazacyclononane-1,4,7-triyltriacetic acid (NOTA), or a derivative thereof.

[0046] In one embodiment, the ligand is added to the reaction mixture in an amount of about 1 μg to about 6000 μg, about 50 μg to about 5000 μg, about 100 μg to about 4500 μg, about 200 μg to about 4000 μg, about 300 μg to about 3000 μg, about 400 μg to about 2000 μg, or about 500 μg to about 1000 μg. In another embodiment, the ligand is about 100 μg, about 200 μg, about 300 μg, about 400 μg, about 500 μg, about 600 μg, about 700 μg, about 800 μg, about 900 μg, about 1000 μg, about 1100 μg, about 1200 μg, about 1300 μg, about 1400 μg, about 1500 μg, about 1600 μg, about 1700 μg, about 1800 μg, about 1900 μg, about 2000 μg, about 2100 μg, about 2200 μg, about 2300 μg, about 2400 μg, about 2500 μg, about 2600 μg, about 2700 μg, about 2800 μg, about 2900 μg, about 3000 μg, about 3100 μg, about 3200 μg, about 3300 μg, about 3400 μg, about 3500 μg, about 3600 μg, about 3700 μg, about 3800 μg, about 3900 μg, about 4000 μg, about 4100 μg, about 4200 μg, about 4300 μg, about 4400 μg, about 4500 μg, about 4600 μg, about 4700 μg, about 4800 μg, about 4900 μg, about 5000 μg, about 5100 μg, about 5200 μg, about 5300 μg, about 5400 μg, about 5500 μg, about 5600 μg, about 5700 μg, The antibody is added to the reaction mixture in an amount of about 2100 μg, about 2200 μg, about 2300 μg, about 2400 μg, about 2500 μg, about 2600 μg, about 2700 μg, about 2800 μg, about 2900 μg, about 3000 μg, about 3100 μg, about 3200 μg, about 3300 μg, about 4400 μg, about 4500 μg, about 5000 μg, about 5500 μg, or about 6000 μg. In yet another embodiment, the ligand is less than about 100 μg, less than about 200 μg, less than about 300 μg, less than about 400 μg, less than about 500 μg, less than about 600 μg, less than about 700 μg, less than about 800 μg, less than about 900 μg, less than about 1000 μg, less than about 1100 μg, less than about 1200 μg, less than about 1300 μg, less than about 1400 μg, less than about 1500 μg, less than about 1600 μg, less than about 1700 μg, less than about 1800 μg, less than about 1900 μg, less than about 200 μg, less than about 300 μg, less than about 350 μg, less than about 400 μg, less than about 450 μg, less than about 460 μg, less than about 470 μg, less than about 480 μg, less than about 490 μg, less than about 500 μg, less than about 510 μg, less than about 520 μg, less than about 530 μg, less than about 540 μg, less than about 550 μg, less than about 5600 μg, less than about 5700 μg, less than about 5800 μg, less than about 5900 μg, less than about 600 μg, less than about 610 μg, less than about 620 μg, less than about 630 μg, less than about 640 μg, less than about 650 μg, less than about 660 μg, less than about 6700 μg, less than about 6800 μg, less than about 6900 μg, less than about 70 In some embodiments, the antibody is added to the reaction mixture in an amount of less than 0 μg, less than 2100 μg, less than about 2200 μg, less than about 2300 μg, less than about 2400 μg, less than about 2500 μg, less than about 2600 μg, less than about 2700 μg, less than about 2800 μg, less than about 2900 μg, less than about 3000 μg, less than about 3100 μg, less than about 3200 μg, less than about 3300 μg, less than about 4400 μg, less than about 4500 μg, less than about 5000 μg, less than about 5500 μg, or less than about 6000 μg.

[0047] In another embodiment, the ligand is used in an amount of about 0.1 ug / mCi to about 20 ug / mCi, about 0.5 ug / mCi to about 15 ug / mCi, about 1 ug / mCi to about 11 ug / mCi, about 1 ug / mCi to about 8 ug / mCi, about 1 ug / mCi to about 5 ug / mCi, about 1 ug / mCi to about 3 ug / mCi, or about 0.1 ug / mCi to about 1.5 ug / mCi. In yet another embodiment, the ligand is used in an amount of about 0.1 ug / mCi, about 0.25 ug / mCi, about 0.4 ug / mCi, about 0.5 ug / mCi, about 0.6 ug / mCi, about 0.75 ug / mCi, about 0.8 ug / mCi, about 1 ug / mCi, about 1.25 ug / mCi, about 1.5 ug / mCi, about 1.75 ug / mCi, about 2 ug / mCi, about 2.5 ug / mCi, about 3 ug / mCi, about 3.5, or about 4 ug / mCi.

[0048] In one embodiment, the concentration of ligand / mL in the radiolabeling step is greater than about 200 ug / mL, greater than about 250 ug / mL, greater than about 300 ug / mL, greater than about 333 ug / mL, or greater than about 400 ug / mL.

[0049] In yet another embodiment, the total labeled ligand is about 200 μg to about 6000 μg, about 500 μg to about 5000 μg, about 1000 μg to about 4000 μg, about 1500 μg to about 3000 μg, about 2000 μg to about 25000 μg, about 2000 μg to about 4000 μg, or about 3000 μg to about 4000 μg. In another embodiment, the total labeled ligand is about 200 μg, about 300 μg, about 400 μg, about 500 μg, about 600 μg, about 700 μg, about 800 μg, about 900 μg, about 1000 μg, about 1100 μg, about 1200 μg, about 1300 μg, about 1400 μg, about 1500 μg, about 1600 μg, about 1700 μg, about 1800 μg, about 1900 μg, about 2000 μg, 2100 μg, about 2200 μg, about 2300 μg, about 2400 μg, about 2500 μg, about 2600 μg, about 2700 μg, about 2800 μg, about 2900 μg, about 3000 μg, about 3100 μg, about 3200 μg, about 3300 μg, about 4000 μg, about 4500 μg, about 5000 μg, about 5500 μg, or about 6000 μg. In yet another embodiment, the total labeled ligand is less than about 500 μg, less than about 1000 μg, less than about 1500 μg, less than about 2000 μg, less than about 2500 μg, less than about 3000 μg, less than about 3500 μg, less than about 4000 μg, less than about 45000 μg, less than about 5000 μg, less than about 5500 μg, less than about 6000 μg, less than about 6500 μg, less than about 7000 μg, less than about 8000 μg, less than about 9000 μg, or less than about 10000 μg. ii. Radionuclides

[0050] In another embodiment, the radionuclide is bismuth-213, chromium-51, cobalt-60, dysprosium-165, erbium-169, holmium-166, iridium-192, iron-59, lead-212, lutetium-177, molybdenum-99, palladium-103, rhenium-186, rhenium-188, samarium-153, strontium-89, tetrahydrofuran-104, tetrahydrofuran-106, tetrahydrofuran-108 ... These include chromium-99m, xenon-133, ytterbium-169, ytterbium-177, yttrium-90, carbon-11, cobalt-57, copper-64, copper-67, fluorine-18, gallium-67, gallium-68, germanium-68, indium-111, rubidium-81, rubidium-82, strontium-82, and thallium-201.

[0051] In another embodiment, 64 CuCl is added to the reaction mixture in an amount of about 100 mCi to about 5000 mCi, about 200 mCi to about 4000 mCi, about 300 mCi to about 3500 mCi, about 400 mCi to about 3000 mCi, about 500 mCi to about 2500 mCi, or up to about 10,000 mCi ( 64 as a source of Cu). In one embodiment, 64 CuCl2 is about 100mCi, about 200mCi, about 300mCi, about 400mCi, about 500mCi, about 600mCi, about 700mCi, about 800mCi, about 900mCi, about 1000mCi, about 1500mCi, about 2000mCi, about 2500mCi, about 3000mCi, about 3500mCi, about 4 about 4,000 mCi, about 4500 mCi, about 5,000 mCi, about 5,500 mCi, about 6,000 mCi, about 6,500 mCi, about 7,000 mCi, about 7,500 mCi, about 8,000 mCi, about 8,500 mCi, about 9,000 mCi, about 9,500 mCi, or about 10,000 mCi 64 In yet another embodiment, 64CuCl2 is less than about 100 mCi, less than about 200 mCi, less than about 300 mCi, less than about 400 mCi, less than about 500 mCi, less than about 600 mCi, less than about 700 mCi, less than about 800 mCi, less than about 900 mCi, less than about 1000 mCi, less than about 1500 mCi, less than about 2000 mCi, less than about 2500 mCi, less than about 3000 mCi, less than about 3500 mCi, is added to the reaction mixture in an amount of less than about 4000 mCi, less than about 4500 mCi, less than about 5000 mCi, less than about 5500 mCi, less than about 6000 mCi, less than about 6500 mCi, less than about 7000 mCi, less than about 7500 mCi, less than about 8000 mCi, less than about 8500 mCi, less than about 9000 mCi, less than about 9500 mCi, or less than about 10,000 mCi ( 64 as a source of Cu).

[0052] In one embodiment, the radionuclide is added to the reaction mixture in an amount of about 0.1 μg, about 0.2 μg, about 0.3 μg, about 0.39 μg, about 0.4 μg, about 0.44 μg, about 0.5 μg, about 0.6 μg, about 0.7 μg, about 0.8 μg, about 0.9 μg, about 1 μg, about 1.12 μg, about 2 μg, about 3 μg, about 4 μg, about 5 μg, about 6 μg, about 7 μg, about 8 μg, about 9 μg, or about 10 μg.

[0053] In yet another embodiment, 64 Cu is added to the reaction mixture in an amount of about 0.1 μg, about 0.2 μg, about 0.3 μg, about 0.4 μg, about 0.44 μg, about 0.5 μg, about 0.6 μg, about 0.7 μg, about 0.8 μg, about 0.9 μg, about 1 μg, about 2 μg, about 3 μg, about 4 μg, about 5 μg, about 6 μg, about 7 μg, about 8 μg, about 9 μg, or about 10 μg. iii.Buffer solution

[0054] In one embodiment, the buffer solution used in preparing the bulk solution of the drug product is sodium acetate buffer, sodium acetate / gentisate buffer, sodium ascorbate buffer, sodium ascorbate / ethanol buffer, ammonium acetate buffer, ammonium acetate / gentisate buffer, ammonium ascorbate buffer, ammonium ascorbate / ethanol buffer, or any other suitable buffer.

[0055] In one embodiment, the concentration of the buffer is about 0.1 M, about 0.2 M, about 0.3 M, about 0.4 M, about 0.5 M, about 0.6 M, about 0.7 M, about 0.8 M, about 0.9 M, or about 1.0 M. In yet another embodiment, the concentration of the buffer is about 20 mg / mL to about 200 mg / mL, about 25 mg / mL to about 190 mg / mL, about 30 mg / mL to about 170 mg / mL, about 35 mg / mL to about 160 mg / mL, about 40 mg / mL to about 150 mg / mL, about 45 mg / mL to about 140 mg / mL, about 45 mg / mL to about 122 mg / mL, about 50 mg / mL to about 130 mg / mL, about 60 mg / mL to about 120 mg / mL, or about 70 mg / mL to about 100 mg / mL. In yet another embodiment, the concentration of the buffer is about 4 mg / mL, about 10 mg / mL, about 20 mg / mL, about 25 mg / mL, about 30 mg / mL, about 35 mg / mL, about 40 mg / mL, about 45 mg / mL, about 50 mg / mL, about 60 mg / mL, about 65% mg / mL, about 66% mg / mL, about 70 mg / mL, about 80 mg / mL, about 90 mg / mL, about 95%, about 100 mg / mL, about 110 mg / mL, about 120 mg / mL, about 122 mg / mL, about 130 mg / mL, about 132 mg / mL, about 140 mg / mL, about 150 mg / mL, about 160 mg / mL, about 170 mg / mL, about 180 mg / mL, about 190 mg / mL, or about 200 mg / mL.

[0056] In another embodiment, the buffer comprises about 4 mg / mL, about 10 mg / mL, about 15 mg / mL, about 20 mg / mL, about 25 mg / mL, about 30 mg / mL, about 35 mg / mL, about 40 mg / mL, about 45 mg / mL, about 50 mg / mL, about 60 mg / mL, about 70 mg / mL, about 80 mg / mL, about 90 mg / mL, or about 100 mg / mL of gentisic acid and about 0.1 M, about 0.2 M, about 0.3 M, about 0.33 M, about 0.4 M, about 0.5 M, about 0.6 M, about 0.7 M, about 0.8 M, about 0.9 M, or about 1.0 M sodium acetate.

[0057] In one specific embodiment, the buffer is a solution of 4 mg / mL gentisic acid and 0.4 M sodium acetate.

[0058] In yet another embodiment, the buffer comprises about 4 mg / mL, about 10 mg / mL, about 20 mg / mL, about 25 mg / mL, about 30 mg / mL, about 35 mg / mL, about 40 mg / mL, about 45 mg / mL, about 46% mg / mL, about 50 mg / mL, about 60 mg / mL, about 64.8 mg / mL, about 66% mg / mL, about 70 mg / mL, about 80 mg / mL, about 90 mg / mL, or about 100 mg / mL of sodium ascorbate and about 1%, about 2%, about 2.8%, about 3%, about 3.5%, about 4%, about 5%, about 8%, about 10%, about 15%, about 20%, about 25%, or about 30% EtOH. In one specific embodiment, the buffer is a solution of 45 mg / mL of sodium ascorbate and 5% EtOH.

[0059] In another embodiment, the buffer comprises about 4 mg / mL, about 10 mg / mL, about 15 mg / mL, about 20 mg / mL, about 25 mg / mL, about 30 mg / mL, about 35 mg / mL, about 40 mg / mL, about 45 mg / mL, about 50 mg / mL, about 60 mg / mL, about 70 mg / mL, about 80 mg / mL, about 90 mg / mL, or about 100 mg / mL of gentisic acid and about 0.1 M, about 0.2 M, about 0.3 M, about 0.33 M, about 0.4 M, about 0.5 M, about 0.6 M, about 0.7 M, about 0.8 M, about 0.9 M, or about 1.0 M sodium ascorbate. iv. Stabilizers

[0060] In one embodiment, the stabilizer is gentisic acid. In another embodiment, the stabilizer is sodium ascorbate. However, any suitable stabilizer may be used.

[0061] In another embodiment, more than one stabilizer is used, hi another embodiment, one stabilizer, such as gentisic acid, is used during the radiolabeling process and another stabilizer, such as sodium ascorbate, is used in the final formulated product.

[0062] In one embodiment, the stabilizer is added in an amount of about 1.0 g to about 9.0 g. In one embodiment, the stabilizer is added in an amount of about 2.0 mg to about 8.0 mg. In another embodiment, the stabilizer is added in an amount of about 3.0 mg to about 7.0 mg. In another embodiment, the stabilizer is added in an amount of about 3.0 mg to about 5.0 mg. In another embodiment, the stabilizer is added in an amount of about 4.0 mg to about 6.0 mg. In one specific embodiment, the stabilizer is added to the reaction mixture in an amount of about 4.0 mg.

[0063] In yet another embodiment, the stabilizer is added in an amount of about 1.0 g to about 9.0 g. In another embodiment, gentisic acid is added in an amount of about 2.0 mg to about 8.0 mg. In another embodiment, gentisic acid is added in an amount of about 3.0 mg to about 7.0 mg. In another embodiment, gentisic acid is added in an amount of about 3.0 mg to about 5.0 mg. In another embodiment, gentisic acid is added in an amount of about 4.0 mg to about 6.0 mg. In one specific embodiment, gentisic acid is added to the reaction mixture in an amount of about 4.0 mg.

[0064] In another embodiment, sodium ascorbate is added in an amount of about 2.0 mg to about 8.0 mg. In another embodiment, sodium ascorbate is added in an amount of about 3.0 mg to about 7.0 mg. In another embodiment, sodium ascorbate is added in an amount of about 3.0 mg to about 5.0 mg. In another embodiment, sodium ascorbate is added in an amount of about 4.0 mg to about 6.0 mg. In one specific embodiment, sodium ascorbate is added to the reaction mixture in an amount of about 4.0 mg. v. Radiolabeling conditions

[0065] In one embodiment, radiolabeling is performed with 500 mCi to 15,000 mCi of radionuclide. The radioactivity concentration of the radiolabel is ≥ 250 mCi / mL, ≥ 300 mCi / mL, ≥ 333 mCi / mL, ≥ 350 mCi / mL, ≥ 400 mCi / mL, ≥ 421 mCi / mL, or ≥ 460 mCi / mL. The total labeled ligand is at a concentration of 1000 to 4000 μg, or ≥ 333 μg / mL.

[0066] In one embodiment, radiolabeling is performed with 500 mCi to 10,000 mCi of radionuclide, the radioactivity concentration of the radiolabel is ≥ 333 mCi / mL, and the total labeled ligand is at a concentration of 1000 to 4000 μg, or ≥ 333 μg / mL.

[0067] In another embodiment, radiolabeling is performed with 500 mCi to 2,500 mCi of radionuclide. The radioactivity concentration of the radiolabel is ≥ 250 mCi / mL, ≥ 300 mCi / mL, ≥ 333 mCi / mL, ≥ 350 mCi / mL, ≥ 400 mCi / mL, ≥ 421 mCi / mL, or ≥ 460 mCi / mL. The total labeled ligand is at a concentration of 1000 to 4000 μg, or ≥ 333 μg / mL.

[0068] In one embodiment, the radiolabel is between 500 mCi and 15,000 mCi. 64The radiolabeling is performed with Cu. The radioactivity concentration is ≥250 mCi / mL, ≥300 mCi / mL, ≥333 mCi / mL, ≥350 μg / mL, ≥400 mCi / mL, ≥421 mCi / mL, or ≥460 mCi / mL. The total labeled DOTATATE concentration is 1000-4000 μg, or ≥333 μg / mL.

[0069] In one embodiment, radiolabeling is performed with 500 mCi to 10,000 mCi of 64Cu. The radioactive concentration of the radiolabel is ≥ 333 mCi / mL. The total labeled DOTATATE is at a concentration of 1000-4000 μg, or ≥ 333 μg / mL.

[0070] In another embodiment, the radiolabel is between 500 mCi and 2,500 mCi. 64 The radiolabeling is performed with Cu. The radioactivity concentration is ≥250 mCi / mL, ≥300 μg / mL, ≥333 mCi / mL, ≥350 μg / mL, ≥400 mCi / mL, ≥421 mCi / mL, or ≥460 mCi / mL. The total labeled DOTATATE is 1000-4000 μg, or a concentration of ≥333 μg / mL.

[0071] In another embodiment, the pH of the reaction mixture is 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, or 7.0. In yet another embodiment, the pH of the reaction mixture is about 4.5 to about 7.0, about 4.6 to about 6.9, about 4.7 to about 6.8, about 4.8 to about 6.7, about 4.9 to about 6.6, about 5.0 to about 6.6, about 5.1 to about 6.5, about 5.2 to about 6.3, about 5.3 to about 6.2, about 5.4 to about 6.1, or about 5.5 to about 6.0. In one specific embodiment, the pH of the reaction mixture is about 5 to about 6.

[0072] Bioconjugate chelates such as DOTA-TATE are generally complexed at temperatures between 50 and 95°C to ensure high radiolabeling and radiolabeling yields. However, the present disclosure provides a method for labeling at lower temperatures, i.e., room temperature or lower, to achieve copper ionization. 64 It is taught that the purity of Cu-DOTATATE is improved due to the more rapid labeling of copper(2+) ions to DOTATATE compared to other common metal impurities.

[0073] In another embodiment, the temperature of the reaction mixture is about 10°C to about 50°C, about 15°C to about 45°C, about 20°C to about 40°C, about 10°C to about 30°C, about 10°C to about 20°C, about 20°C to about 50°C, about 20°C to about 40°C, or about 20°C to about 30°C. In one embodiment, the temperature of the reaction mixture is about 10°C, about 15°C, about 20°C, about 22°C, about 25°C, about 30°C, about 35°C, about 40°C, about 45°C, or about 50°C. In another embodiment, the temperature of the reaction mixture is ambient temperature.

[0074] In yet another embodiment, the temperature of the reaction mixture is less than or equal to 50°C, less than 50°C, less than or equal to 45°C, less than 45°C, less than or equal to 40°C, less than 40°C, less than or equal to 35°C, less than 35°C, less than or equal to 30°C, less than 30°C, less than or equal to 25°C, less than 25°C, less than or equal to 20°C, less than 20°C, less than or equal to 15°C, less than 15°C, less than or equal to 10°C, or less than 10°C.

[0075] In one embodiment, the molar ratio of ligand to radionuclide in the reaction mixture is about 125:1, 120:1, 115:1, 110:1, 105:1, 100:1, 95:1, 90:1, 85:1, 80:1, 75:1, 70:1, 65:1, 60:1, 55:1, 50:1, 45:1, 40:1, 35:1, 30:1, 25:1, 20:1, 15:1, 10:1, 5:1, 4:1, 3:1, 2:1, 2.5:1, or 1:1. In another embodiment, the molar ratio of ligand to radionuclide in the reaction mixture is from about 125:1 to about 75:1. In yet another embodiment, the molar ratio of ligand to radionuclide in the reaction mixture is from about 105:1 to about 95:1. In yet another embodiment, the molar ratio of ligand to radionuclide in the reaction mixture is from about 110:1 to about 90:1. In yet another embodiment, the molar ratio of ligand to radionuclide in the reaction mixture is from about 102:1 to about 99:1. In yet another embodiment, the molar ratio of ligand to radionuclide in the reaction mixture is from about 125:1 to about 1:1, from about 105:1 to about 10:1, from about 102:1 to about 10:1, from about 110:1 to about 50:1, from about 90:1 to about 70:1, or from about 60:1 to about 1:1, or from about 110:1 to about 90:1.

[0076] In one specific embodiment, DOTATATE 64 The molar ratio of DOTATATE to Cu is about 125:1, 120:1, 115:1, 110:1, 105:1, 100:1, 95:1, 90:1, 85:1, 80:1, 75:1, 70:1, 65:1, 60:1, 55:1, 50:1, 45:1, 40:1, 35:1, 30:1, 25:1, 20:1, 15:1, 10:1, 5:1, 4:1, 3:1, 2:1, 2.5:1, or 1:1. 64 In yet another embodiment, the molar ratio of DOTATATE to Cu in the reaction mixture is from about 105:1 to about 95:1. 64 In yet another embodiment, the molar ratio of DOTATATE to Cu in the reaction mixture is from about 102:1 to about 99:1. 64The molar ratio to Cu is about 125:1 to about 1:1, about 105:1 to about 10:1, about 102:1 to about 10:1, about 110:1 to about 50:1, about 90:1 to about 70:1, or about 60:1 to about 1:1, or about 110:1 to about 90:1.

[0077] In one embodiment, the ratio of ligand mass (μg):radionuclide activity (mCi) is about 5:1, 4:1, 3:1, 2:1, or 1:1. In yet another embodiment, the concentration of ligand mass (μg) to radionuclide activity (mCi) is about 1.0 μg / mCi, 0.9 μg / mCi, 0.8 μg / mCi, 0.7 μg / mCi, 0.6 μg / mCi, 0.5 μg / mCi, 0.4 μg / mCi, 0.3 μg / mCi, 0.2 μg / mCi, or 0.1 μg / mCi for each reaction. In yet another embodiment, the concentration of ligand mass (μg) to radionuclide activity (mCi) was about 0.6 μg / mCi for each reaction.

[0078] In one embodiment, the mass of the ligand (μg): 64 The ratio of radioactivity (mCi) of Cu to Cu is about 5:1, 4:1, 3:1, 2:1, or 1:1. 64 The concentration of ligand mass (μg) to Cu radioactivity (mCi) is about 1.0 μg / mCi, 0.9 μg / mCi, 0.8 μg / mCi, 0.7 μg / mCi, 0.6 μg / mCi, 0.5 μg / mCi, 0.4 μg / mCi, 0.3 μg / mCi, 0.2 μg / mCi, or 0.1 μg / mCi for each reaction. 64 The concentration of ligand mass (μg) to Cu radioactivity (mCi) was approximately 0.6 μg / mCi for each reaction.

[0079] In one embodiment, the mass (μg) of DOTATATE is: 64 The ratio of radioactivity (mCi) of Cu to Cu is about 5:1, 4:1, 3:1, 2:1, or 1:1. 64The concentration of DOTATATE mass (μg) relative to Cu radioactivity (mCi) is about 1.0 μg / mCi, 0.9 μg / mCi, 0.8 μg / mCi, 0.7 μg / mCi, 0.6 μg / mCi, 0.5 μg / mCi, 0.4 μg / mCi, 0.3 μg / mCi, 0.2 μg / mCi, or 0.1 μg / mCi for each reaction. 64 The concentration of DOTATATE mass (μg) relative to Cu radioactivity (mCi) was approximately 0.6 μg / mCi for each reaction.

[0080] In one embodiment, the radioactivity of the bulk solution of the drug substance is from about 1 mCi to about 10,000 mCi, from about 1 mCi to about 9,900 mCi, from about 1 mCi to about 9,800 mCi, from about 1 mCi to about 9,700 mCi, from about 1 mCi to about 9,600 mCi, from about 1 mCi to about 9,500 mCi, from about 1 mCi to about 9,400 mCi, from about 1 mCi to about 9,300 mCi, from about 1 mCi to about 9,200 mCi, from about 1 mCi to about 9,100 mCi, from about 1 mCi to about 9,000 mCi, from about 1 mCi to about 8,900 mCi, from about 1 mCi to about 8,800 mCi, from about 1 mCi to about 8,700 mCi, 1mCi ~ approx. 8,600mCi, approx. 1mCi ~ approx. 8,500mCi, approx. 1mCi ~ approx. 8,400mCi, approx. 1mCi ~ approx. 8,300mCi , about 1mCi to about 8,200mCi, about 1mCi to about 8,100mCi, about 1mCi to about 8,000mCi, about 1mCi to about 7,900m Ci, about 1mCi to about 7,800mCi, about 1mCi to about 7,700mCi, about 1mCi to about 7,600mCi, about 1mCi to about 7,50 0mCi, about 1mCi to about 7,400mCi, about 1mCi to about 7,300mCi, about 1mCi to about 7,200mCi, about 1mCi to about 7, 100mCi, approximately 1mCi to approximately 7,000mCi, approximately 1mCi to approximately 6,900mCi, approximately 1mCi to approximately 6,800mCi, approximately 1mCi to approximately 6,700mCi, approximately 1mCi to approximately 6,600mCi, approximately 1mCi to approximately 6,500mCi, approximately 1mCi to approximately 6,400mCi, approximately 1mCi ~6,300mCi, approximately 1mCi~6,200mCi, approximately 1mCi~6,100mCi, approximately 1mCi~6,000mCi, approximately 1m Ci ~ approx. 5,900mCi, approx. 1mCi ~ approx. 5,800mCi, approx. 1mCi ~ approx. 5,700mCi, approx. 1mCi ~ approx. 5,600mCi, approx. 1mCi to approx. 5,500mCi, approx. 1mCi to approx. 5,400mCi, approx. 1mCi to approx. 5,300mCi, approx. 1mCi to approx. 5,200mCi , about 1mCi to about 5,100mCi, about 1mCi to about 5,000mCi, about 1mCi to about 4,900mCi, about 1mCi to about 4,800m Ci, approximately 1mCi to approximately 4,700mCi, approximately 1mCi to approximately 4,600mCi, approximately 1mCi to approximately 4,500mCi, approximately 1mCi to approximately 4,40 0mCi, about 1mCi to about 4,300mCi, about 1mCi to about 4,200mCi, about 1mCi to about 4,100mCi, about 1mCi to about 4,000mCi, approximately 1mCi to approximately 3,900mCi, approximately 1mCi to approximately 3,800mCi, approximately 1mCi to approximately 3,700mCi, approximately 1mCi to approximately 3,600mCi, approximately 1mCi to approximately 3,500mCi, approximately 1mCi to approximately 3,400mCi, approximately 1mCi to approximately 3 ,300mCi, about 1mCi to about 3,200mCi, about 1mCi to about 3,100mCi, about 1mCi to about 3000mCi, about 10mCi to about 2900mCi, about 20mCi to about 2,800mCi, about 30mCi to about 2,700mCi, about 40mCi to about 2,600 mCi, about 50 mCi to about 2,500 mCi, about 60 mCi to about 2,400 mCi, about 70 mCi to about 2,300 mCi, about 80 mCi to about 2,200 mCi, about 90 mCi to about 2,100 mCi, about 100 mCi to about 2,000 mCi, about 150 mCi to about 3,000 mCi, about 200 mCi to about 2,500 mCi, about 250 mCi to about 2,000 mCi, about 300 mCi to about 1,500 mCi, about 400 mCi to about 1,000 mCi, or about 500 mCi to about 750 mCi.

[0081] In another embodiment, the radioactivity of the bulk solution of the drug substance is about 1 mCi, about 20 mCi, about 40 mCi, about 60 mCi, about 80 mCi, about 100 mCi, about 120 mCi, about 140 mCi, about 160 mCi, about 200 mCi, about 220 mCi, about 240 mCi, about 260 mCi, about 280 mCi, about 300 mCi, about 320 mCi, about 340 mCi, about 360 mCi, about 380 mCi, about 400 mCi, about 420 mCi, about 440 mCi, about 460 mCi, about 480 mCi, about 500 mCi, about 550 mCi, about 600 mCi, about 650 mCi, about 700 mCi , about 750mCi, about 800mCi, about 850mCi, about 900mCi, about 950mCi, about 1,000mCi, about 1,100mCi, about 1,200mCi, approximately 1,300mCi, approximately 1,400mCi, approximately 1,500mCi, approximately 1,600mCi, approximately 1,700mCi, approximately 1, 800mCi, approx. 1,900mCi, approx. 2,000mCi, approx. 2,100mCi, approx. 2,200mCi, approx. 2,300mCi, approx. 2,40 0mCi, approx. 2,500mCi, approx. 2,600mCi, approx. 2,700mCi, approx. 2,800mCi, approx. 2,900mCi, approx. 3,000m Ci, approx. 3,100mCi, approx. 3,200mCi, approx. 3,300mCi, approx. 3,400mCi, approx. 3,500mCi, approx. 3,600mCi , approx. 3,700mCi, approx. 3,800mCi, approx. 3,900mCi, approx. 4,000mCi, approx. 4,100mCi, approx. 4,200mCi, approx. 4,300mCi, approx. 4,400mCi, approx. 4,500mCi, approx. 4,600mCi, approx. 4,700mCi, approx. 4,800mCi, approx. 4, 900mCi, approx. 5,000mCi, approx. 5,100mCi, approx. 5,200mCi, approx. 5,300mCi, approx. 5,400mCi, approx. 5,50 0mCi, approx. 5,600mCi, approx. 5,700mCi, approx. 5,800mCi, approx. 5,900mCi, approx. 6,000mCi, approx. 6,100m Ci, approx. 6,200mCi, approx. 6,300mCi, approx. 6,400mCi, approx. 6,500mCi, approx. 6,600mCi, approx. 6,700mCi , approx. 6,800mCi, approx. 6,900mCi, approx. 7,000mCi, approx. 7,100mCi, approx. 7,200mCi, approx. 7,300mCi, approx. 7,400mCi, approximately 7,500mCi, approximately 7,600mCi, approximately 7,700mCi, approximately 7,800mCi, approximately 7,900mCi, approximately 8,000mCi, about 8,100mCi, about 8,200mCi, about 8,300mCi, about 8,400mCi, about 8,500mCi, about 8,600mCi, about 8,700mCi, about 8,800mCi, about 8,900mCi, about 9,000mCi, 9,100mCi, about 9,200mCi, about 9,300mCi, about 9,400mCi, about 9,500mCi, about 9,600mCi, about 9,700mCi, about 9,800mCi, about 9,900mCi, or about 10,000mCi. In one specific embodiment, the radioactivity of the bulk solution of drug substance is about 100 mCi, 500 mCi, 1000 mCi, 2000 mCi, 3,000 mCi, 4,000 mCi, 5,000 mCi, 6,000 mCi, 7,000 mCi, 8,000 mCi, 9,000 mCi, or 10,000 mCi.

[0082] In one embodiment, the volume of the radionuclide solution is about 0.1 mL to about 10 mL, about 0.2 mL to about 9 mL, about 0.3 mL to about 8 mL, about 0.4 mL to about 7 mL, about 0.5 mL to about 6 mL, about 1 mL to about 5 mL, or about 2 mL to about 4 mL. In another embodiment, the volume of the radionuclide solution is about 0.1 mL, about 0.2 mL, about 0.3 mL, about 0.4 mL, about 0.5 mL, about 1 mL, about 2 mL, about 3 mL, about 4 mL, about 5 mL, about 6 mL, about 7 mL, about 8 mL, about 9 mL, or about 10 mL. In one embodiment, 64 The volume of the Cu solution is about 0.1 mL to about 10 mL, about 0.2 mL to about 9 mL, about 0.3 mL to about 8 mL, about 0.4 mL to about 7 mL, about 0.5 mL to about 6 mL, about 1 mL to about 5 mL, or about 2 mL to about 4 mL. 64 The volume of the Cu solution is about 0.1 mL, about 0.2 mL, about 0.3 mL, about 0.4 mL, about 0.5 mL, about 1 mL, about 2 mL, about 3 mL, about 4 mL, about 5 mL, about 6 mL, about 7 mL, about 8 mL, about 9 mL, or about 10 mL.

[0083] In yet another embodiment, the volume of the radiolabeled solution is about 0.1 mL to about 10 mL, about 0.5 mL to about 9 mL, about 1 mL to about 7 mL, about 1.5 mL to about 6 mL, about 0.5 mL to about 6 mL, about 1 mL to about 5 mL, or about 2 mL to about 4 mL. In another embodiment, the volume of the radiolabeled solution is about 0.1 mL, about 0.2 mL, about 0.3 mL, about 0.4 mL, about 0.5 mL, about 1 mL, about 1.5 mL, about 2 mL, about 3 mL, about 4 mL, about 5 mL, about 6 mL, about 7 mL, about 8 mL, about 9 mL, or about 10 mL.

[0084] In another embodiment, the reaction time is about 1 minute, about 2 minutes, about 3 minutes, about 4 minutes, about 5 minutes, about 6 minutes, about 7 minutes, about 8 minutes, about 9 minutes, about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, about 30 minutes, about 45 minutes, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, or about 10 hours. In one embodiment, the reaction time is about 1 minute to about 24 hours, about 1 minute to about 18 hours, about 1 minute to about 12 hours, or about 1 minute to about 6 hours. In yet another embodiment, the reaction time is about 1 minute to about 60 minutes, about 2 minutes to about 45 minutes, or about 5 minutes to about 30 minutes.

[0085] In yet another embodiment, the concentration of the antiradiolytic agent in the final formulation is 29-122 mg / mL + 1-5% ethanol.

[0086] In yet another embodiment, the amount of non-radioactive copper added to the reaction mixture is 0-30 μg / mL (ppm). In yet another embodiment, the amount of non-radioactive copper added to the reaction mixture is 0.1-30 μg / mL (ppm). D. 64 Purification of Cu-DOTATATE bulk solution

[0087] In yet another embodiment, the bulk metal-ligand solution is purified using a C-18 Light Sep Pak or any suitable purification system / column.

[0088] In one embodiment, the elution solvent in the purification is ethanol, 5% ethanol (95% water), 10% ethanol (90% water), 15% ethanol (85% water), 20% ethanol (80% water), 25% ethanol (75% water), 30% ethanol (70% water), 35% ethanol (65% water), 40% ethanol (60% water), 45% ethanol (55% water), 50% ethanol (50% water), 55% ethanol (45% water), 60% ethanol (40% water), 65% ethanol (35% water), 70% ethanol (30% water), 75% ethanol (25% water), 80% ethanol (20% water), 85% ethanol (15% water), 90% ethanol (10% water), 95% ethanol (5% water), or 100% ethanol.

[0089] In yet another embodiment, the volume of solvent from the purification step is about 0.1 mL to about 10 mL, about 0.5 mL to about 9 mL, about 1 mL to about 7 mL, about 1.5 mL to about 6 mL, about 0.5 mL to about 6 mL, about 1 mL to about 5 mL, or about 2 mL to about 4 mL. In another embodiment, the volume of solvent from the purification step is about 0.1 mL, about 0.2 mL, about 0.3 mL, about 0.4 mL, about 0.5 mL, about 1 mL, about 1.5 mL, about 2 mL, about 3 mL, about 4 mL, about 5 mL, about 6 mL, about 7 mL, about 8 mL, about 9 mL, or about 10 mL.

[0090] In another embodiment, the drug product 64 The radionuclide purity of Cu is ≥99%, ≥99.1%, ≥99.2%, ≥99.3%, ≥99.4%, ≥99.5%, ≥99.6%, ≥99.7%, ≥99.8%, or ≥99.9%.

[0091] In yet another embodiment, the amount of radionuclide impurities in the drug product is ≦1%, ≦0.9%, ≦0.8%, ≦0.7%, ≦0.6%, ≦0.5%, ≦0.4%, ≦0.3%, ≦0.2%, or ≦0.1%. In another embodiment, the amount of one single radionuclide impurity in the drug product is ≦0.1%, ≦0.09%, ≦0.08%, ≦0.07%, ≦0.06%, ≦0.05%, ≦0.04%, ≦0.03%, ≦0.02%, or ≦0.01%.

[0092] In one embodiment, the radiochemical purity of the drug product is ≧90%, ≧91%, ≧92%, ≧93%, ≧94%, ≧95%, ≧96%, ≧97%, ≧98%, ≧99%, ≧99.1%, ≧99.2%, ≧99.3%, ≧99.4%, ≧99.5%, ≧99.6%, ≧99.7%, ≧99.8%, or ≧99.9% as copper Cu64 DOTATATE.

[0093] In one embodiment, purity is measured using high performance liquid chromatography (HPLC) or any other acceptable or suitable technique.

[0094] In another embodiment, gentisic acid is present in the drug product in an amount of ≦50 ppm, ≦40 ppm, ≦30 ppm, ≦20 ppm, ≦10 ppm, ≦5 ppm, or ≦1 ppm.

[0095] In one embodiment, a single impurity is present in an amount of ≦1%, ≦0.9%, ≦0.8%, ≦0.7%, ≦0.6%, ≦0.5%, ≦0.4%, ≦0.3%, ≦0.2%, or ≦0.1% of DOTATATE and related substances in the drug product.

[0096] In another embodiment, total impurities are present in an amount of ≦10%, ≦9%, ≦8%, ≦7%, ≦6%, ≦5%, ≦4%, ≦3%, ≦2%, ≦1%, ≦0.9%, ≦0.8%, ≦0.7%, ≦0.6%, ≦0.5%, ≦0.4%, ≦0.3%, ≦0.2%, or ≦0.1% of DOTATATE and related substances in the drug product.

[0097] In another embodiment, the bacterial endotoxin is present in the drug product in an amount of ≦100 EU / mL, ≦90 EU / mL, ≦80 EU / mL, ≦70 EU / mL, ≦60 EU / mL, ≦50 EU / mL, ≦40 EU / mL, ≦39 EU / mL, ≦30 EU / mL, ≦20 EU / mL, ≦10 EU / mL, ≦9 EU / mL, ≦8 EU / mL, ≦7 EU / mL, ≦6 EU / mL, ≦5 EU / mL, ≦4 EU / mL, ≦3 EU / mL, ≦2 EU / mL, or ≦1 EU / mL. F. Drug Products (64 Cu-DOTATATE injection)

[0098] The drug product disclosed herein is indicated for use with positron emission tomography (PET) for the localization of somatostatin receptor-positive neuroendocrine tumors (NETs) in adult patients. i. Chemical characteristics

[0099] The drug product described herein is a copper ion implantable radioactive diagnostic agent for use in PET imaging. 64 It contains Cu-DOTATATE. Chemically, 64 Cu-DOTATATE is described as copper (Cu64)-N-[(4,7,10-tricarboxymethyl-1,4,7,10-tetraazacyclododec-1-yl)acetyl]-diphenylalanyl-L-cysteinyl-L-tyrosyl-D-tryprophanyl-L-lysyl-L-threoninyl-L-cysteinyl-L-threonine-cyclic (2-7) disulfide. It has a molecular weight of 1497.2 daltons, and the following is the structural formula of one isomeric form: [ka]

[0100] The drug product is a sterile, clear, colorless to yellow solution for intravenous use. Each 10 mL single-dose vial contains 148 MBq (4 mCi) of 64 Cu-DOTATATE is contained in a 4 mL solution volume at the calibration date. Additionally, each mL of solution contains 40 mg of ascorbic acid, 0.05 mL of dehydrated alcohol, USP (ethanol) in sterile water for injection, USP. The pH is adjusted with sodium hydroxide and hydrochloric acid to be between approximately 5.5 and 7.5. ii. Physical characteristics

[0101] Tables 1 and 2 are 64 Provides the primary radiation emission data and physical attenuation of Cu. 64 Cu has a half-life of t 1 / 2 = 12.7 hours: (a) 17.6% 64(b) Positron emission into Ni, resulting in the emission of two 511 keV annihilation photons (35.7%). 64 38.5% by beta decay to Zn, and (c) 64 The decay of Cu-64 also results in the emission of a characteristic 1346 keV gamma ray with an intensity of about 0.48%.

[0102] The gamma emission spectrum of the drug product exhibits peaks at about 511 keV and about 1346 keV. [Table 1] [Table 2] iii. External radiation

[0103] Gamma constant: 3.6 x 10 per MBq at 1 meter -5 mSv / hour (0.133 mrem / hour per mCi at 1 meter). Table 3 64 Radiation attenuation of Cu by lead shielding is shown. [Table 3]

[0104] In one embodiment, the drug product is stored at a temperature of about 15° C. to about 30° C., about 15° C. to about 25° C., about 15° C. to about 20° C., or about 20° C. to about 30° C. In another embodiment, the drug product is stored at a temperature of about 10° C., about 15° C., about 20° C., about 22° C., about 25° C., or about 30° C. In yet another embodiment, the drug product is stored at a controlled room temperature of about 20° C. to about 25° C.

[0105] In another embodiment, the drug product is stored at a temperature of about 30°C to about 60°C, about 35°C to about 55°C, about 40°C to about 50°C, or about 50°C to about 60°C. In another embodiment, the drug product is stored at a temperature of about 30°C, about 35°C, about 40°C, about 45°C, about 50°C, about 55°C, or about 60°C. In yet another embodiment, the drug product is stored at a temperature of about 50°C to about 55°C.

[0106] Radiochemical identity can be confirmed using HPLC. 64 The HPLC relative retention time (RRT) of Cu-DOTATATE correlates with that of a DOTATATE standard. 64 The HPLC RRT of Cu-DOTATATE is about 1 to about 2, or about 1.15 to about 1.25.

[0107] Radionuclide identity can be confirmed using gamma emission spectroscopy. The gamma emission spectrum of the drug product exhibits peaks at about 511 keV and about 1346 keV.

[0108] In one embodiment, the solution volume of the drug product is about 1 mL to about 10 mL, about 2 mL to about 9 mL, about 3 mL to about 7 mL, about 4 mL to about 6 mL, or about 3 mL to about 6 mL. In yet another embodiment, the solution volume of the drug product is about 1 mL, about 2 mL, about 3 mL, about 4 mL, about 5 mL, about 6 mL, about 7 mL, about 8 mL, about 9 mL, or about 10 mL.

[0109] In yet another embodiment, the drug product contains 148 MBq (4 mCi) (37 MBq (1 mCi) per mL) at the calibration date and time. 64 A sterile, clear, colorless to yellow solution in a single-dose vial containing Cu-DOTATATE. The sealed vial is placed in a shielded (lead) container for radiation protection. The drug product is shipped in Type A packaging.

[0110] In one embodiment, the total vial radioactivity (assay) is from about 1.0 mCi / vial to about 10 mCi / vial, from about 1.5 mCi / vial to about 9 mCi / vial, from about 2.0 mCi / vial to about 8 mCi / vial, from about 2.5 mCi / vial to about 7 mCi / vial, from about 3.0 mCi / vial to about 6 mCi / vial, or from about 3.6 mCi / vial to about 4.4 mCi / vial. In another embodiment, the total vial radioactivity (assay) is about 1.0 mCi / vial, about 1.5 mCi / vial, about 2.0 mCi / vial, about 2.5 mCi / vial, about 3.0 mCi / vial, about 3.5 mCi / vial, about 3.6 mCi / vial, about 4.0 mCi / vial, about 4.4 mCi / vial, about 4.5 mCi / vial, about 5.0 mCi / vial, about 5.5 mCi / vial, about 6 mCi / vial, about 7 mCi / vial, about 8 mCi / vial, about 9 mCi / vial, or about 10 mCi / vial.

[0111] In yet another embodiment, the radioactivity concentration of the drug product is from about 0.5 mCi / mL to about 15 mCi / mL, from about 0.5 mCi / mL to about 12.5 mCi / mL, from about 0.5 mCi / mL to about 10 mCi / mL, from about 0.5 mCi / mL to about 7.5 mCi / mL, from about 0.5 mCi / mL to about 5 mCi / mL, from about 0.5 mCi / mL to about 3 mCi / mL, from about 0.6 mCi / mL to about 2.5 mCi / mL, from about 0.7 mCi / mL to about 2.0 mCi / mL, from about 0.8 mCi / mL to about 1.5 mCi / mL, or from about 0.9 mCi / mL to about 1.1 mCi / mL. In yet another embodiment, the radioactivity concentration of the drug product is about 15 mCi / mL, about 14 mCi / mL, about 13 mCi / mL, about 12 mCi / mL, about 11 mCi / mL, about 10 mCi / mL, about 9 mCi / mL, about 8 mCi / mL, about 7 mCi / mL, about 6 mCi / mL, or about 5 mCi / mL. In another embodiment, the radioactivity concentration of the drug product is about 5-15 mCi / mL. In a further embodiment, the radioactivity concentration of the drug product is about 9-14 mCi / mL. In yet another embodiment, the radioactivity concentration of the drug product is about 10-11 mCi / mL. In yet another embodiment, the radioactivity concentration of the drug product is about 11-12 mCi / mL. In an additional embodiment, the radioactivity concentration of the drug product is about 12-13 mCi / mL.

[0112] In another embodiment, DOTATATE and related substances are present in the drug product in an amount of ≦50 ppm, ≦40 ppm, ≦30 ppm, ≦27 ppm, ≦22.7 ppm, ≦20 ppm, or ≦10 ppm.

[0113] In one embodiment, the apparent specific activity of the drug product, upon calibration, is ≧10 mCi / mg, ≧20 mCi / mg, ≧30 mCi / mg, ≧40 mCi / mg, ≧50 mCi / mg, ≧60 mCi / mg, ≧70 mCi / mg, ≧80 mCi / mg, or ≧90 mCi / mg of DOTATATE and related substances.

[0114] In another embodiment, the average specific radioactivity of the drug product is about 2.96 MBq / μg. In another embodiment, the average specific radioactivity of the drug product is about 1.0 to about 5.0 MBq / μg. In another embodiment, the average specific radioactivity of the drug product is about 2.0 to about 4.0 MBq / μg. In another embodiment, the average specific radioactivity of the drug product is about 2.5 to about 3.5 MBq / μg. In yet another embodiment, the average specific activity of the drug product, upon calibration, is about 0.5 MBq / μg, about 1.0 MBq / μg, about 1.5 MBq / μg, about 2.0 MBq / μg, about 2.5 MBq / μg, about 3.0 MBq / μg, about 3.5 MBq / μg, about 4.0 MBq / μg, about 4.5 MBq / μg, about 5.0 MBq / μg, about 6.0 MBq / μg, about 7.0 MBq / μg, about 8.0 MBq / μg, about 9.0 MBq / μg, or about 10.0 MBq / μg.

[0115] In yet another embodiment, the fill volume of the drug product in the vial is about 1 mL to about 10 mL, about 2 mL to about 8 mL, about 3 mL to about 6 mL, or about 3.6 mL to about 4.4 mL. In yet another embodiment, the fill volume of the drug product in the vial is about 1 mL, about 2 mL, about 3 mL, about 3.6 mL, about 4 mL, about 4.4 mL, about 5 mL, about 6 mL, about 7 mL, about 8 mL, about 9 mL, or about 10 mL.

[0116] In another embodiment, the pH of the drug product is about 4.5 to about 8.0, about 4.6 to about 7.9, about 4.7 to about 7.8, about 4.8 to about 7.7, about 4.9 to about 7.6, about 5.0 to about 7.5, or about 5.5 to about 7.5. In another embodiment, the pH of the drug product is 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, or 7.5.

[0117] In one embodiment, the content uniformity of the drug product is ≦10%, ≦9%, ≦8%, ≦7%, ≦6%, ≦5%, ≦4%, ≦3%, ≦2%, ≦1%, ≦0.9%, ≦0.8%, ≦0.7%, ≦0.6%, ≦0.5%, ≦0.4%, ≦0.3%, ≦0.2%, or ≦0.1%.

[0118] In another embodiment, ethanol is present in the drug product in an amount of about 1% to about 10%, about 2% to about 9%, about 3% to about 8%, about 4% to about 7%, or about 4% to about 6%. In yet another embodiment, ethanol is present in the drug product in an amount of about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, or about 10%.

[0119] In one embodiment, the ascorbic acid content in the drug product is about 1 mg / mL to about 100 mg / mL, about 10 mg / mL to about 90 mg / mL, about 20 mg / mL to about 80 mg / mL, about 3 mg / mL to about 70 mg / mL, about 40 mg / mL to about 60 mg / mL, about 30 mg / mL to about 60 mg / mL, or about 36 mg / mL to about 44 mg / mL. In another embodiment, the ascorbic acid content in the drug product is about 1 mg / mL, about 10 mg / mL, about 15 mg / mL, about 20 mg / mL, about 30 mg / mL, about 36 mg / mL, about 40 mg / mL, about 44 mg / mL, about 50 mg / mL, about 60 mg / mL, about 70 mg / mL, about 80 mg / mL, about 90 mg / mL, or about 100 mg / mL.

[0120] In one embodiment, the RCP of the drug product is ≧90%, ≧91%, ≧92%, ≧93%, ≧94%, ≧95%, ≧96%, ≧97%, ≧98%, or ≧99%.

[0121] In another embodiment, the drug product has a resolved radiochemical yield (RCY) of about 50%, about 55%, about 56%, about 60%, about 65%, about 68%, about 70%, about 75%, about 80%, about 83%, about 85%, about 90%, or about 95% (decay corrected).

[0122] In one embodiment, a filter integrity test is performed on the drug product. In another embodiment, the drug product is tested for sterility.

[0123] iv. Dosage of drug product

[0124] With respect to drug product dose as used herein, the present disclosure provides a drug product dose sufficient to enable positron emission tomography (PET) imaging in a subject in need thereof. 64 An effective amount of Cu-DOTATATE is provided.

[0125] In one embodiment, the dose of drug product administered to a subject in need thereof is from about 20 MBq to about 350 MBq, from about 30 MBq to about 340 MBq, from about 40 MBq to about 330 MBq, from about 50 MBq to about 320 MBq, from about 60 MBq to about 310 MBq, from about 70 MBq to about 300 MBq, from about 80 MBq to about 290 MBq, from about 90 MBq to about 280 MBq, from about 100 MBq to about 270 MBq, from about 110 MBq to about 260 MBq, from about 132 MBq to about 163 MBq, or from about 111 MBq to about 185 MBq, or from about 120 MBq to about 250 MBq at the calibration date and time.

[0126] In another embodiment, the dose of drug product administered to a subject in need thereof is about 20MBq, about 30MBq, about 37MBq, about 40MBq, about 50MBq, about 60MBq, about 70MBq, about 80MBq, about 90MBq, about 100MBq, about 110MBq, about 111MBq, about 120MBq, about 130MBq, about 140MBq, about 148MBq, about 150MBq, about 160MBq, about 170MBq, about 180MBq, about 190MBq, about 200MBq, about 210MBq, about 220MBq, about 230MBq, about 240MBq, about 250MBq, about 260MBq, about 270MBq, about 280MBq, about 290MBq, about 300MBq, about 310MBq, about 320MBq, about 330MBq, about 340MBq, about 350MBq, about 360MBq, about 370MBq, about 380MBq, about 390MBq, about 400MBq, about 410MBq, about 420MBq, about 430MBq, about 440MBq, about 450MBq, about 460MBq, about 470MBq, about 480MBq, about 490MBq, about 500MBq, about 510MBq, about 520MBq, about 530MBq, about 540MBq, about 550MBq, about 560MBq, about 570MBq, about 580MBq, about 590MBq, about 600MBq, about 610MBq MBq, about 170 MBq, about 180 MBq, about 185 MBq, about 190 MBq, about 200 MBq, about 210 MBq, about 220 MBq, about 230 MBq, about 240 MBq, about 250 MBq, about 260 MBq, about 270 MBq, about 280 MBq, about 290 MBq, about 300 MBq, about 310 MBq, about 320 MBq, about 330 MBq, about 340 MBq, or about 350 MBq.

[0127] In one embodiment, the dose of drug product administered to a subject in need thereof is from about 0.5 mCi to about 9.5 mCi, from about 0.54 mCi to about 9.0 mCi, from about 0.6 mCi to about 8.5 mCi, from about 0.7 mCi to about 8 mCi, from about 0.8 mCi to about 7.5 mCi, from about 0.9 mCi to about 7 mCi, from about 1.0 mCi to about 6.5 mCi, from about 1.1 mCi to about 6 mCi, about 1.2 mCi to about 5.5 mCi, about 1.3 mCi to about 5.0 mCi, about 1.4 mCi to about 4.5 mCi, about 1.5 mCi to about 4.0 mCi, about 2 mCi to about 3 mCi, about 0.1 mCi to about 10 mCi, about 0.5 mCi to about 5 mCi, about 1 mCi to about 5 mCi, or about 1 mCi to about 4 mCi.

[0128] In yet another embodiment, the dose of drug product administered to a subject in need thereof is about 0.1 mCi, about 0.5 mCi, about 0.54 mCi, about 0.6 mCi, about 0.7 mCi, about 0.8 mCi, about 0.9 mCi, about 1.0 mCi, about 1.1 mCi, about 1.2 mCi, about 1.3 mCi, about 1.4 mCi, about 1.5 mCi, about 2.0 mCi, about 2.5 mCi, about 3.0 mCi, about 3.1 mCi, about 3.2 mCi, about 3.3 mCi, about 3.4 mCi, about 3.5 mCi, or about 3.6 mCi at the calibration date and time. i, about 3.6 mCi, about 3.7 mCi, about 3.8 mCi, about 3.9 mCi, about 4.0 mCi, about 4.1 mCi, about 4.2 mCi, about 4.3 mCi, about 4.4 mCi, about 4.5 mCi, about 4.6 mCi, about 4.7 mCi, about 4.8 mCi, about 4.9 mCi, about 5.0 mCi, about 5.5 mCi, about 6.0 mCi, about 6.5 mCi, about 7.0 mCi, about 7.5 mCi, about 8.0 mCi, about 8.5 mCi, about 9.0 mCi, about 9.5 mCi, or about 10.0 mCi.

[0129] In one embodiment, the dose of the drug product is administered intravenously. In another embodiment, the drug product is introduced intravenously to a subject in need thereof in a single dose, two doses, three doses, or multiple doses.

[0130] In one particular embodiment, the drug product is administered to the subject as an intravenous bolus injection at a dose of about 148 MBq (or about 4 mCi), and images are acquired about 45 to about 90 minutes after drug administration.

[0131] Dose selection for elderly patients should be conservative, usually starting at the lower end of the dosing range, reflecting the greater frequency of decreased liver, renal, or cardiac function and coexisting or other medications.

[0132] In one embodiment, the drug product is administered to the subject over a period of about 15 minutes, about 10 minutes, about 5 minutes, about 4 minutes, about 3 minutes, about 2 minutes, or about 1 minute.

[0133] In one particular embodiment, the amount of radioactivity to be administered for PET imaging in adults is 148 MBq (4 mCi) administered as an intravenous injection over a period of approximately 1 minute.

[0134] In one specific embodiment, the drug product is in a single dose vial at the time of calibration. 64 Contains 148 MBq (4 mCi) of Cu-DOTATATE at a concentration of 37 MBq (1 mCi) per mL. v. Imaging

[0135] Somatostatin analogues include: 64 Cu-DOTATATE competitively binds to the same somatostatin receptors and may interfere with imaging. Patients are imaged immediately before receiving a somatostatin analog. For patients receiving a long-acting somatostatin analog, a 28-day washout period is recommended before imaging. For patients receiving a short-acting somatostatin analog, a 2-day washout period is recommended before imaging.

[0136] For drug product PET imaging, a whole-body acquisition from crown to mid-thigh is recommended. Image acquisition begins approximately 45 to 90 minutes after intravenous drug product administration. Drug product uptake time and scan duration are adapted according to the equipment used and patient and tumor characteristics to obtain optimal image quality. G. Method of administration of drug products

[0137] 64 Cu-DOTATATE binds to somatostatin receptors. Based on the intensity of the signal, 64 PET images obtained using Cu-DOTATATE injection demonstrate the presence and density of somatostatin receptors in tissue. Uptake can also be seen in various non-NET tumors that contain somatostatin receptors, or as a normal physiological variation. NET tumors that do not have somatostatin receptors are not visualized.

[0138] The method of administering the drug product to the patient is: (a) 64 calibrating the Cu-DOTATATE injection; (b) Within approximately 2 hours after the calibration time 64 using Cu-DOTATATE injections; (c) 64 using aseptic technique and radiation shielding when discontinuing and administering Cu-DOTATATE injections; (d) visually for particulate matter and discoloration before administration; 64 inspecting the Cu-DOTATATE injection and using it only if the solution is free of particulate matter and does not discolor; (e) calculating the volume required to administer based on the measured radioactivity, volume, calibration time, and date; (f) measuring the patient's dose immediately prior to administration of the drug product using a dose calibrator; (g) 64 administering to the patient an intravenous flush of 0.9% Sodium Chloride Injection, USP after the injection of Cu-DOTATATE injection; and (h) Disposing of any unused medication in a safe manner in accordance with applicable regulations. Includes.

[0139] 64The estimated radiation absorbed dose per injected radioactivity for organs and tissues in adult patients after intravenous administration of Cu-DOTATATE injection is shown in Table 4. [Table 4]

[0140] The effective radiation dose resulting from the administration of 148 MBq (4 mCi) to an adult is approximately 4.7 mSv. At an administered activity of 148 MBq (4 mCi), typical radiation doses to the critical organs—liver, kidney / adrenal glands, and spleen—are approximately 24 mGy, 21 mGy, and 17 mGy, respectively. Because the spleen has one of the highest physiological uptakes, higher uptakes and radiation doses to other organs or pathological tissues may occur in patients who have undergone splenectomy.

[0141] Non-radioactive somatostatin analogues and 64 Cu-DOTATATE competitively binds to the somatostatin receptor (SSTR2). Patients are imaged immediately prior to administration of the somatostatin analog. For patients receiving long-acting somatostatin analogs, a 28-day washout period is recommended before imaging. For patients receiving short-acting somatostatin analogs, a 2-day washout period is recommended before imaging.

[0142] 64 Cu-DOTATATE uptake reflects the level of somatostatin receptor density in NETs, ​​but uptake can also be seen in a variety of other tumors that also express somatostatin receptors. Increased uptake can also be seen in other non-cancerous pathological conditions that express somatostatin receptors, including thyroid disease, or in subacute inflammation, or can occur as a normal physiological anomaly (e.g., the uncinate process of the pancreas).

[0143] A negative scan after administration of a drug product in a patient with no history of NET disease does not rule out disease.

[0144] 64One to three hours after a single dose of Cu-DOTATATE injection, maximum radioactivity is observed in the adrenal glands, kidneys, pituitary gland, spleen, and liver.

[0145] 64 After a single intravenous dose of Cu-DOTATATE injection (4.15 ± 0.13 mCi) (n = 6), between 16% and 40% of the injected dose of radioactivity was recovered in urine over a 6-hour collection period.

[0146] In one embodiment, 64 After a single intravenous administration of Cu-DOTATATE injection, about 5%, about 10%, about 15%, about 16%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, or about 50% of the radioactivity of the injected dose is recovered in the urine over a 6-hour collection period.

[0147] In another embodiment, 64 After a single intravenous administration of Cu-DOTATATE injection, about 5%, about 10%, about 15%, about 16%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, or about 50% of the radioactivity of the injected dose is recovered in the urine over a 5-hour collection period.

[0148] In yet another embodiment, 64 After a single intravenous dose of Cu-DOTATATE injection, about 5%, about 10%, about 15%, about 16%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, or about 50% of the radioactivity of the injected dose is recovered in the urine over a 4-hour collection period.

[0149] In one embodiment, 64 After a single intravenous administration of Cu-DOTATATE injection, about 5%, about 10%, about 15%, about 16%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, or about 50% of the radioactivity of the injected dose is recovered in the urine over a 3-hour collection period.

[0150] In another embodiment, 64After a single intravenous administration of Cu-DOTATATE injection, about 5%, about 10%, about 15%, about 16%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, or about 50% of the radioactivity of the injected dose is recovered in the urine over a 2-hour collection period.

[0151] In yet another embodiment, 64 After a single intravenous administration of Cu-DOTATATE injection, about 5%, about 10%, about 15%, about 16%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, or about 50% of the radioactivity of the injected dose is recovered in the urine over a 1-hour collection period. [Example]

[0152] The following examples are based on high purity 64 An improved process for producing Cu-labeled DOTATATE is provided. By labeling DOTATATE with copper at low temperatures (i.e., ≦30° C.), chelation of other metals by DOTATATE can be reduced, thereby providing a drug product of higher purity. Furthermore, these examples demonstrate the ability to produce a drug product of higher purity while maintaining sufficient chemical stability for distribution to patients. 64 To provide a useful process for scaling up the production of Cu-DOTATATE, the following buffer solutions were prepared for use in Examples 1-5 described below.

[0153] Sodium acetate / gentisic acid buffer: Gentisic acid (GA) and sodium acetate (NaOAc) were dissolved in high resistance water (HRW), and the pH of the resulting solution was adjusted using glacial acetic acid or 1 M sodium hydroxide. The solution was further diluted with HRW to achieve the desired concentrations of NaOAc and GA.

[0154] Sodium ascorbate buffer: Sodium ascorbate was dissolved in HRW. The pH of the solution was adjusted to 6.5-7.5 using 1M HCl or 1M NaOH. The solution was further diluted with HRW to achieve the desired concentration of sodium ascorbate.

[0155] Sodium ascorbate / ethanol buffer: Sodium ascorbate was dissolved in HRW and absolute ethanol. The pH was adjusted to 6.5-7.5 with either 1 M HCl or 1 M NaOH, then further diluted with HRW to achieve the desired final concentrations of both sodium ascorbate and ethanol.

[0156] HPLC was performed using an Agilent 1200 Series system (Bioscan B-FC-200P) equipped with a variable wavelength UV-Vis detector followed in-line by a sodium iodide detector. A Phenomenex Luna C18 column (150 mm x 4.6 mm, 5 μm) was used. The mobile phase consisted of solvent A and solvent B. Solvent A was 0.1% trifluoroacetic acid (TFA) in HRW, and solvent B was 0.1% TFA acid in acetonitrile (ACN). The gradient was (i) 15–40% solvent B in A for 10 min, 40% solvent B in A for 10–15 min, 15% solvent B in A for 15–16 min, and 15% solvent B in A for 16–19 min. The flow rate was 1.2 mL / min, and UV detection was monitored at 220 nm.

[0157] Example 1 Preparation of non-radioactive Cu-DOTATATE

[0158] An initial non-radioactive reaction was performed to prepare Cu-DOTATATE by mixing a solution of CuCl in 0.05 M HCl with a solution of DOTATATE peptide in a gentisic acid / sodium acetate buffer solution. The pH of the buffer solution was 6 unless otherwise noted. Formation of Cu-DOTATATE was confirmed via HPLC. The relative retention times of the DOTATATE starting material, the Cu-DOTATATE product peak, and other reaction components were established. A representative chromatogram is shown in Figure 2 for a standard solution containing gentisic acid and DOTATATE.

[0159] In initial experiments, approximately equimolar amounts of DOTATATE (0.035 μmol) in NaOAc / GA buffer and copper metal cation (0.039 μmol) in 0.05 M HCl were mixed in a vial at room temperature. The reaction mixture was analyzed by HPLC at multiple time intervals. The HPLC chromatogram for the sample collected at 5 min, presented in Figure 3, showed a new product peak identified as Cu-DOTATATE at approximately 7.2 min, which corresponded to the reduction of the DOTATE precursor peak (retention time approximately 6 min). The data suggest that the formation of Cu-DOTATATE is rapid and quantitative in reaction mixtures containing equimolar amounts of starting materials at ambient temperature.

[0160] Example 2 Formation of metal dotate complexes favors copper over other common metals

[0161] Due to the high specific activity of Cu-64, a typical 64 Cu]Cu 2+ In solution, nano- to microgram amounts of Cu 2+ There will be a 64 Cu]Cu 2+ Other trace metals that may be present in the solution are typically environmental impurities introduced by the manufacturing process. Common transition metals that may be present include iron, lead, zinc, and nickel. The effect that metallic impurities have on the preparation of copper Cu64 Dotatate is unknown in these experiments because they are chemically identical. 64 Instead of Cu nat A non-radioactive solution of Cu was used for the evaluation.

[0162] A 0.05 M HCl solution containing 0.44 μg (0.00692 μmol) of Cu was mixed with 100 μg (0.0693 μmol) of DOTATATE in a sodium acetate / gentisate buffer at room temperature (approximately 22 °C) to obtain a molar ratio of DOTATATE to Cu of 10:1. The reaction was monitored by HPLC. As shown in Table 5, the HPLC peak areas of the DOTATATE and Cu-DOTATE compounds showed essentially no change in the peak areas obtained at 5 minutes and 7 hours, indicating that the formation of Cu-DOTATATE was rapid at room temperature and was complete after 5 minutes. [Table 5]

[0163] isotopically enriched 64 Ni is typically 64 Ni is another potential metallic impurity, as it is used in the production of Cu. However, because it is difficult to achieve HPLC baseline separation between Ni-DOTATATE and Cu-DOATATE, similar competition experiments were performed using Ni instead of Cu to evaluate the reaction kinetics of Ni. Non-radiolabeling reactions were performed by mixing a solution of Ni (0.0063 μmol), Fe (0.0069 μmol), Zn (0.0067 μmol), and Co (0.0068 μmol) in 0.05 M HCl with a solution of DOTATATE (0.0693 μmol) in gentisic acid / sodium acetate buffer at room temperature (approximately 22 °C). To better evaluate the chelation behavior of Ni in the presence of Fe, Zn, and Co, copper was not added to the solution.

[0164] The reaction mixture was analyzed by HPLC at both 5 min and 6 h. The peak area results are summarized in Table 6. 2+ The reaction kinetics of Cu with DOTATATE 2+ Thus, similar to other transition metals (e.g., Fe, Co, Zn), Cu chelation by DOTATATE appears to occur faster than Ni at ambient temperatures. [Table 6]

[0165] Example 3 Formation of Cu-DOTATATE at lower temperatures

[0166] An experiment similar to that described in Example 2 was performed to determine whether Cu-DOTATATE formation would similarly occur at reduced temperatures (i.e., 15° C. to 18° C.). Reaction mixtures were prepared as described in Example 2, but adjusted as necessary to meet the conditions outlined in Table 7. [Table 7]

[0167] Each reaction mixture was sampled for HPLC analysis after approximately 5 minutes. The reaction mixture containing only Cu was also sampled after approximately 2 hours. The results are summarized in Table 8. These data confirm that labeling of DOTATATE is largely complete after 5 minutes, even in the presence of Fe at reduced temperatures. [Table 8]

[0168] Example 4 Up to 2,000 mCi in a single reaction 64 Preparation of Cu-DOTATATE

[0169] The radiolabeling reaction was carried out according to the general procedure described herein. 64 A solution of Cu in 0.05M HCl was mixed with a solution of DOTATATE in sodium acetate / gentisate buffer. The reaction mixture was heated to 30°C for 5 minutes and then purified via a C-18 solid-phase extraction cartridge. 64Cu-DOTATATE was collected in 2 mL of 50% EtOH and then diluted with ascorbic acid solution. The final product was assayed for radioactivity and radiochemical purity (RCP) was assessed via radio-HPLC analysis. Characterization was performed using non-radioactive DOTATATE (t R = 6.3 min) and Cu-DOTATATE (t R = 7.3 min) was performed against a standard.

[0170] Generally, scale-up reactions from 100 mCi to approximately 7,000 mCi of Cu-64 were achieved for a single reaction. Representative reactions and results for batch sizes from 100 mCi to 7000 mCi are outlined in Table 9. HPLC analysis showed that the major product peak had a retention time (t) of approximately 7.4 minutes. R ), which is non-radioactive Cu-DOTATATE (t R = 7.3 min). R The remaining time (6.2-7.2 min) is due to decomposition products resulting from radiolysis. The chemical stability of the purified reaction solution was monitored by HPLC and, as shown in Table 9, 64 Cu-DOTATATE was demonstrated to be stable over a period of at least 47 hours. [Table 9]

[0171] Example 5 Greater than 7,500 mCi 64 Preparation of Cu-DOTATATE

[0172] Higher batch sizes 64 Cu-DOTATATE can be prepared by combining two sub-batches. For example, 5,250 mCi 64 Cu(R1) and 4,800 mCi 64Two radiolabeling reactions consisting of Cu(R2) were performed to prepare a total of approximately 9 Ci of copper Cu64 dotatate (decay not corrected for time of synthesis). 64 The Cu radiolabeling reaction was carried out in 0.05M HCl. 64 Cu, a solution of DOTATATE in gentisic acid / sodium ascorbate buffer, 64 The purified drug product solutions from each radiolabeling reaction were combined and diluted to a total of approximately 9 Ci of DOTATATE at the time of synthesis, at a ratio of 0.6 μg of DOTATATE per mCi of Cu. 64 Cu-DOTATATE was obtained upon purification. The process yield was ≥ 95%. The RCP of the final drug product solution upon release was ≥ 96%.

[0173] Example 6 The maximum concentration of DOTATATE was 64 Cu-DOTATATE batch

[0174] 64 Before the preparation of Cu-DOTATATE, the DOTATATE ligand was added to the reaction mixture. 64 Cu was added at a ratio of 1 μg DOTATATE per mCi (i.e., approximately a 170:1 molar ratio at the time of synthesis). 64 To improve the molar activity of Cu-DOTATATE, process improvements were initiated to reduce the amount of DOTATATE in the radiolabeling reactions. Two radiolabeling reactions, reaction 1 (R1) and reaction 2 (R2), were performed using up to 5,250 mCi each. 64 Cu and up to 4,800mCi 64 Cu. In the case of R1, the total labeled DOTATATE was approximately 3,125 μg or a concentration of ≧276 μg / mL. In the case of R2, the total labeled DOTATATE was approximately 3,018 μg or a concentration of ≧265 μg / mL. The ratio of the radionuclide (i.e., moles of DOTATATE) to the ligand (i.e., moles of DOTATATE) in the reaction mixture was 0.018 μg. 64 The molar ratios of SiO to Cu (moles of SiO) were about 102:1 (R1) and about 99:1 (R2). 64The concentration of ligand mass (µg) relative to Cu radioactivity (mCi) was approximately 0.6 µg / mCi for each reaction. 64 The Cu solution was combined with a solution of DOTATATE in sodium acetate / gentisate buffer. The radioactivity concentrations (RAC) of R1 and R2 were ≥460 mCi / mL and ≥421 mCi / mL, respectively. The reactions were held at 30°C for 5 minutes and at ambient temperature for 5 minutes before purification.

[0175] The crude reaction mixtures for R1 and R2 were each purified using a C-18 solid phase extraction (SPE) cartridge, and the combined eluates containing the purified product yielded approximately 8.7 Ci of 64 A bulk solution containing Cu-DOTATATE was prepared (decay was not corrected during synthesis). The process yield was ≥95%, and the radiochemical purity (RCP) of the final drug product solution after dilution was ≥96%.

[0176] In another experiment, approximately 7 Ci 64 Cu-DOTATATE was prepared in a single reaction using a ratio of 0.6 μg of DOTATATE per mCi of Cu-64 (see Table 9).

[0177] Example 7 64 Effect of gentisic acid and ethanol on Cu-DOTATATE product stability

[0178] 64 The effect of ethanol (EtOH) and gentisic acid (GA) content in the final dose matrix on the chemical stability of Cu-dotatate was evaluated. In these experiments, 500 mCi reactions were performed using the general procedure outlined in Example 2. 64 The purified Cu-DOTATATE product was eluted from the Sep-Pak with 2 mL of 50% EtOH(aq) into 5 mL of 50 mg / mL NaOAsc buffer. 64 This was used as a Cu-DOTATATE stock solution. 64From the Cu-DOTATATE stock, 1 mL aliquots were transferred to vials containing the following solutions: [Table 1A]

[0179] Each of the vials was analyzed for stability by HPLC, and the results are summarized in Table 10. Surprisingly, the highest amount of degradation (24%-38%) was in those samples that contained high amounts of GA, which is generally considered to be a radioprotectant. 64 The reduction of Cu-DOTATATE is R = Release at 6.6 and 7.1 min 64 The decomposition correlated with increases in Cu and two unknown radioactive impurities. Since vials with similar activity concentrations showed little or no decomposition, it is unlikely that the decomposition resulted from radiolysis, and therefore the mechanism leading to chemical instability remains unknown. 64 The only other condition that resulted in more than a 2% loss of Cu-DOTATATE was vial 3, which differed from the control (vial 1) in that it contained approximately 10% EtOH. [Table 10]

[0180] The results showed that the radioactivity of the serotonin-containing solution was greater than 95% pure in solutions containing sodium ascorbate at concentrations ranging from 45 to 122 mg / mL, ethanol concentrations ranging from 1.6% to 5.2%, and radioactivity concentrations (as prepared) ranging from 3.6 to 16 mCi / mL. 64 It was shown that Cu-DOTATATE remained chemically stable for 2 days. 64 The Cu-dotatate product remained stable with greater than 90% purity for solutions containing sodium ascorbate concentrations up to 98 mg / mL and ethanol contents up to 9.7%, with radioactivity concentrations of approximately 18 mCi / mL (as prepared).

[0181] Example 8 Preparation of Cu-DOTATATE in the presence of increasing gentisic acid or sodium ascorbate

[0182] The general reaction scheme used in previous experiments was repeated, except that the concentration of gentisic acid in the reaction mixture was increased four-fold. After the reaction, the mixture was sampled and purified via a C-18 solid-phase extraction (SPE) cartridge, and the purified product was analyzed via HPLC to determine the reaction yield. The results for the HPLC analysis are summarized in Table 11. Near-quantitative recoveries of DOTATATE and Cu-DOTATATE were achieved, and neither the labeling efficiency nor the purification was affected by the excessive excess of gentisic acid in the reaction mixture. [Table 11]

[0183] Gentisic acid in the reaction mixture acts as a radioprotectant, helping to reduce radiolytic degradation. To evaluate the possibility of using an alternative radioprotectant, reactions were performed in which sodium ascorbate was added to the reaction mixture (pH = 6.8). DOTATATE (0.0693 μmol) in sodium acetate / gentisic acid buffer was added to Cu 2+ in 0.05 M HCl (0.0069 μmol) and diluted with sodium ascorbate to give Cu(II) of DOTATATE. 2+ The ratio of Cu-DOTATATE to Cu was adjusted to 10:1. The reaction mixture was mixed at room temperature, and samples were taken at 5 and 51 minutes to monitor the formation of Cu-DOTATATE via HPLC analysis. The HPLC peak areas for Cu-DOTATATE were 2.33 mV / min at 5 minutes and 2.36 mV / min at 51 minutes, indicating that the reaction was complete by 5 minutes.

[0184] Example 9 64 Effect of non-radioactive copper on the radiochemical purity of Cu-DOTATATE

[0185] Three reactions, each using approximately 5 Ci, were performed. R1 was a control reaction without the addition of non-radioactive copper. No copper was detected in R1. Non-radioactive copper was added to R2 and R3 to investigate its effect on RCP. R2 had a total copper content of approximately 139 μg (11.0 μg / mL) in 12.6 mL. R3 had a total copper content of approximately 476 μg (31.1 μg / mL) in 15.3 mL. The total DOTATATE labeled in each reaction was approximately 3000 μg (R1), approximately 3000 μg (R2), and approximately 3600 μg (R3), or concentrations of ≥250 μg / mL, ≥238 μg / mL, and ≥235 μg / mL, respectively. The reaction time for each reaction was approximately 5 minutes. After heating, each reaction was cooled to room temperature for approximately 5 minutes, and the mixture was subsequently refined and diluted to its final bulk solution.

[0186] The final drug product solutions of R1, R2, and R3 had RACs of approximately 11.7 mCi / mL (R1), approximately 10.3 mCi / mL (R2), and approximately 12.4 mCi / mL (R3), respectively. The decay-corrected process yields of R1, R2, and R3 were approximately 95.4%, approximately 98.7%, and approximately 95.6%, respectively. The RCPs of each final drug product solution after dilution were ≥95.5% (R1), ≥97.3% (R2), and ≥97.9% (R3).

[0187] Example 10 Recovery of DOTATATE from SPE cartridges at higher flow rates

[0188] Typically, the flow rate through the SPE cartridge is low (i.e., 1-5 mL / min) to ensure adequate loading of the desired product onto the cartridge and high recovery of purified product eluate. 64In the case of Cu-DOTATATE, concentrating the product on an SPE cartridge can result in higher radiolytic damage, especially in high-activity batches. Therefore, to reduce purification time, DOTATATE recovery was evaluated at higher flow rates. Because C-18 SPE chemistry is primarily driven by DOTATATE interaction with the cartridge, experiments were performed using non-radioactive solutions of DOTATATE, as the behavior of Cu-DOTATATE or other metal-DOTATATE species is very similar.

[0189] A solution of DOTATATE in sodium acetate / gentisate buffer was prepared and loaded onto a C-18 SPE cartridge at a flow rate of either 12 mL / min or 18 mL / min. The cartridge was rinsed with water, and DOTATATE was eluted in 50% EtOH. The amount of DOTATATE in the load solution and purified product eluate was assessed via HPLC. At a flow rate of 12 mL / min, 5.3% of the DOTATATE broke through the SPE cartridge during loading, while 97.1% was recovered in the eluate (total recovery of 102%). When purification was performed at 18 mL / min, 28.4% of the DOTATATE broke through during loading, while 68.7% was recovered in the eluate (total recovery of 97%). Results suggest that flow rates of at least up to 12 mL / min can be maintained while maintaining near-quantitative recovery yields. 64 This demonstrates that the method can be used to purify Cu-DOTATATE. The loading solution was collected in fractions, and the DOTATATE recovery results for individual fractions are presented in Figures 4 and 5. Example 11 Improved purification yield of Cu-DOTATATE using 50% ethanol eluent

[0190] Typically, radiolabeled copper Cu-64 Dotatate is purified by C-18 SPE. In this procedure, the crude radiolabeled solution is loaded onto a C-18 SPE cartridge, the cartridge is rinsed with water to remove hydrophilic impurities, and then the purified copper is purified. 64Cu-DOTATATE compounds are typically eluted from cartridges using 100% ethanol. We found that the purification yield of Cu-DOTATATE can be improved by using 50% EtOH. Several experiments were performed to confirm this observation.

[0191] Reaction mixtures containing copper(2+) ions, transition metal ion impurities, and bioconjugate chelator (DOTATATE) were prepared in triplicate for each condition. The reaction mixtures were held at room temperature (approximately 20°C) for 5 minutes and then purified using C-18 SPE cartridges and eluted with either 100% EtOH (n=3) or 50% EtOH (n=3). The ratios of DOTATATE to metal in the reaction mixtures are presented in Table 12. [Table 12]

[0192] Approximately 10 minutes after mixing, samples of the crude reaction mixtures were analyzed by HPLC to obtain in situ reaction yields. Each reaction mixture was then purified through a C-18 SPE cartridge, and the product was eluted with either 100% EtOH or 50% EtOH. The isolated yield of the purified product solution was determined by HPLC analysis. The results are presented in Table 13. Reaction yields were determined by comparison with a DOTATATE standard. [Table 13]

[0193] Example 12 Drug product effectiveness

[0194] The efficacy of the drug product was established in two single-center, open-label studies. Study 1 prospectively evaluated a total of 63 subjects, including 42 patients with known or suspected NETs based on histology, conventional imaging, or clinical evaluation, and 21 healthy volunteers. Of the 42 patients, 37 (88%) had a history of NETs at the time of drug product imaging. Of the total study population of 63 subjects, 28 (44%) were male and 35 (56%) were female, with most subjects being Caucasian (86%). The mean age of subjects was 54 years (range, 25-82 years).

[0195] Drug product images from each subject were interpreted as either positive or negative for NETs by three independent readers blinded to clinical information and other imaging results. PET imaging results were compared with a single oncologist's blinded assessment of subject diagnosis based on available histopathology results, reports of conventional imaging performed within 8 weeks prior to drug product imaging (MRI, contrast-enhanced CT, bone scintigraphy, [ 18 F]fluorodeoxyglucose PET / CT, 18 F] sodium fluoride PET / CT, 111 In]Indium pentetreotide SPECT / CT, 68 The results were compared to a composite reference standard consisting of [Ga]Ga-doatate PET / CT) and clinical and laboratory data, including chromogranin A and serotonin levels. The proportion of subjects positive for disease per composite reference identified as positive by drug product imaging was used to quantify positive agreement. The proportion of subjects without disease per composite reference identified as negative by drug product imaging was used to quantify negative agreement. Table 14 shows the performance of drug products in detecting NETs for Study 1. [Table 14]

[0196] Study 2 demonstrated similar performance through a retrospective analysis of published data collected on 112 patients (63 men, 49 women; mean age 62 years, range 30–84 years) with a known history of NETs.

[0197] Example 13 Safety and efficacy of drug products

[0198] In the safety and efficacy study, 71 subjects received a single dose of the drug product. Of these 71 subjects, 21 were healthy volunteers and the remainder were patients with known or suspected NETs. The following adverse reactions occurred at a rate of <2%: (a) gastrointestinal disorders: nausea, vomiting; and (b) vascular disorders: flushing.

[0199] 126 patients with a known history of NETs 64 A single injection of Cu-DOTATATE was administered. Four patients reported experiencing nausea immediately after injection.

[0200] The embodiments described herein are intended to be exemplary only, and those skilled in the art will recognize that variations and modifications may be made without departing from the scope of the invention, which is encompassed by the claims that follow. In one embodiment, for example, the following items are provided: (Item 1) 1. A method for radiolabeling DOTATATE, comprising: Reacting copper-64 with a buffered solution containing DOTATATE Includes; The reaction occurs in less than 15 minutes at a temperature less than or equal to 30°C; The method wherein the molar ratio of DOTATATE to copper-64 in the reaction solution is from about 110:1 to about 90:1. (Item 2) 2. The method for radiolabeling DOTATATE according to item 1, wherein the reaction occurs in less than 10 minutes. (Item 3) 2. The method for radiolabeling DOTATATE according to item 1, wherein the reaction occurs in about 5 minutes. (Item 4) 2. The method for radiolabeling DOTATATE according to item 1, wherein the molar ratio of DOTATATE to copper-64 in the reaction solution is about 100:1. (Item 5) 2. The method for radiolabeling DOTATATE according to item 1, wherein the reaction occurs at a temperature below 25°C. (Item 6) 2. The method for radiolabeling DOTATATE according to item 1, wherein the reaction occurs at a temperature below 20°C. (Item 7) 2. The method for radiolabeling DOTATATE according to item 1, wherein the reaction occurs at a temperature below 15°C. (Item 8) Prepared by the method described in Item 1 64 Drug products containing Cu-DOTATATE. (Item 9) 64 1. A method for preparing a drug product comprising Cu-DOTATATE, comprising: The method, wherein the drug product is prepared by (i) radiolabeling DOTATATE with copper-64 at a concentration of about 0.6 μg / mL (μg of DOTATATE per mCi of copper-64), and the radionuclide purity of the copper-64 in the drug product is about 99%. (Item 10) Prepared by the method described in Item 9 64 Drug products containing Cu-DOTATATE. (Item 11) of DOTATATE 64 10. The method of claim 9, wherein the molar ratio of SiO to Cu is about 125:1, 120:1, 115:1, 110:1, 105:1, 100:1, 95:1, 90:1, 85:1, 80:1, 75:1, 70:1, 65:1, 60:1, 55:1, 50:1, 45:1, 40:1, 35:1, 30:1, 25:1, 20:1, 15:1, 10:1, 5:1, 4:1, 3:1, 2:1, 2.5:1, or 1:1. (Item 12) 10. The method of claim 9, wherein the drug product has a resolved radiochemical yield (RCY) of about 50%, about 55%, about 56%, about 60%, about 65%, about 68%, about 70%, about 75%, about 80%, about 83%, about 85%, about 90%, or about 95% (decay corrected). (Item 13) 10. The method of claim 9, wherein the radiolabeling is achieved in about 1 minute, about 2 minutes, about 3 minutes, about 4 minutes, about 5 minutes, about 6 minutes, about 7 minutes, about 8 minutes, about 9 minutes, about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, about 30 minutes, about 45 minutes, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 15 hours, about 20 hours, or about 22 hours. (Item 14) 10. The method of claim 9, wherein the radiolabeling is achieved in less than 15 minutes. (Item 15) Item 10. The method according to Item 9, wherein the radiolabeling is carried out at a reaction temperature of about 10°C to about 50°C, about 15°C to about 45°C, about 20°C to about 40°C, about 10°C to about 30°C, about 10°C to about 20°C, about 20°C to about 50°C, about 20°C to about 40°C, or about 20°C to about 30°C. (Item 16) 10. The method according to item 9, wherein the radiolabeling is carried out at a pH of about 4.5 to about 7.0, about 4.6 to about 6.9, about 4.7 to about 6.8, about 4.8 to about 6.7, about 4.9 to about 6.6, about 5.0 to about 6.6, about 5.1 to about 6.5, about 5.2 to about 6.3, about 5.3 to about 6.2, about 5.4 to about 6.1, or about 5.5 to about 6.0. (Item 17) 1. A method for radiolabeling DOTATATE, comprising: Reacting copper-64 with a buffered solution containing DOTATATE Includes; The reaction occurs at a temperature less than or equal to 30°C in less than 15 minutes; and DOTATATE was added at a concentration of approximately 0.6 μg / mL ( 64 The method can be radiolabeled with copper-64 at 1 μg of DOTATATE per mCi of Cu. (Item 18) Prepared by the method described in Item 17 64 Drug products containing Cu-DOTATATE. (Item 19) 18. The method of claim 17, wherein non-radioactive copper is added to the reaction mixture. (Item 20) 20. The method according to item 19, wherein 0.1 to 30 μg / mL (ppm) of non-radioactive copper is added to the reaction mixture. (Item 21) 64 1. A drug product for use in positron emission tomography, comprising Cu-DOTATATE, 64 Cu-DOTATATE, 148MBq 64 1. A drug product stored in a single-dose vial containing Cu-DOTATATE, wherein said drug product has a radioactivity concentration of about 5-15 mCi / mL and wherein said drug product after dilution has a radiochemical purity of ≥ 96%. (Item 22) 22. The drug product of item 21, wherein the radiochemical purity of the drug product after dilution is ≧97%. (Item 23) 22. The drug product of item 21, wherein the radiochemical purity of the drug product after dilution is ≧98%. (Item 24) 22. The drug product of item 21, wherein the radiochemical purity of the drug product after dilution is ≧99%. (Item 25) 22. The drug product of item 21, wherein the drug product is stable for 48 hours after formulation. (Item 26) 22. The drug product of item 21, wherein the drug product is stable for 24 hours after formulation. (Item 27) 22. The drug product according to item 21, wherein the radioactivity concentration of the drug product is about 9 to 14 mCi / mL. Product. (Item 28) 22. The drug product according to item 21, wherein the radioactivity concentration of the drug product is about 10 to 11 mCi / mL.

Claims

1. ６４ 1. A drug product for use in positron emission tomography comprising Cu-DOTATATE ( 64 Cu—N-[(4,7,10-tricarboxymethyl-1,4,7,10-tetraazacyclododec-1-yl)acetyl]-(D)-phenylalanyl-(L)-cysteinyl-(L)-tyrosyl-(D)-tryptophanyl-(L)-lysyl-(L)-threoninyl-(L)-cysteinyl-(L)-threonine-cyclic disulfide (Cys 2 -Cys 7 )), said drug product comprising a molar ratio of 125:1 to 1:1 total DOTATATE:total 64 Cu, and containing 148 MBq (4 mCi) of total DOTATATE in a 4 mL solution volume at the time of calibration. ６４ A drug product, which is a single-dose vial containing Cu-DOTATATE, wherein said drug product has a radiochemical purity of ≧96%.

2. The drug product described in claim 1, wherein the radiochemical purity of the drug product is ≧97%.

3. The drug product described in claim 1, wherein the radiochemical purity of the drug product is ≧98%.

4. The drug product described in claim 1, wherein the radiochemical purity of the drug product is ≧99%.

5. 10. The drug product of claim 1, wherein the drug product is stable for 48 hours after formulation.

6. 10. The drug product of claim 1, wherein the drug product is stable for 24 hours after formulation.

7. The drug product of claim 1, wherein the molar ratio of total DOTATATE:total 64 Cu is from 125:1 to 75:1.

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