Radiolabeling and preparations used for 64Cu-DOTATATE amplification

Abstract

This disclosure relates to a method for creating a robust procedure capable of providing commercially available radiodiagnostic agents suitable for use in conjunction with positron emission tomography (PET) for the localization of somatostatin receptor-positive neuroendocrine tumors (NETs) in adult patients.

Description

[0001] Cross-references to related applications

[0002] This application claims priority to U.S. Provisional Application No. 63 / 074,451, filed September 3, 2020, the entire contents of which are incorporated herein by reference in their entirety to the extent permitted by law. Technical Field

[0003] This disclosure relates to radiolabeling and purification. 64 Compositions and methods for Cu-DOTATATE 64 Cu-DOTATATE is a bioconjugate compound containing a radioactive nuclide that emits positrons. Background Technology

[0004] Known imaging techniques of paramount importance in medical diagnosis include positron emission tomography (PET), computed tomography (CT), magnetic resonance imaging (MRI), single-photon computed tomography (SPECT), and ultrasound (US). Although current imaging techniques have advanced significantly, they primarily rely on nonspecific, macroscopic, physical, physiological, or metabolic changes that distinguish pathological tissues from normal tissues.

[0005] Targeting molecular imaging (MI) has the potential to usher in a new dimension in medical diagnostics. The term "targeting" relates to the selective and highly specific binding of natural or synthetic ligands (adhesives) to molecules of interest (molecular targets) in vitro or in vivo.

[0006] Medical imaging (MI) is a rapidly emerging biomedical research discipline that can be defined as the visualization, characterization, and quantification of biological processes at the cellular and subcellular levels within a complete organism. It is a new multidisciplinary field where the resulting images reflect the in vivo mechanisms and cellular and molecular pathways of disease in the physiological real-world environment, rather than identifying the molecular events that lead to disease.

[0007] Several different contrast enhancers are currently known. They can be used for functional imaging and are primarily being developed for PET and SPECT. The application of radiolabeled bioactive peptides in diagnostic imaging is becoming increasingly important in nuclear medicine. Bioactive molecules that selectively interact with specific cell types can be used to deliver radioactivity to target tissues. For example, radiolabeled peptides have great potential for delivering radionuclides to tumors, infarcts, and infected tissues for diagnostic imaging and radiotherapy.

[0008] DOTA (1,4,7,10-tetrakis(carboxymethyl)-1,4,7,10-tetraazacyclo dodecane) and its derivatives constitute an important class of chelating agents for biomedical applications because they are very stable in accommodating a variety of divalent and trivalent metal ions. One of the derivatives of DOTA is DOTATATE, [(4,7,10-tricarboxymethyl-1,4,7,10-tetraazacyclododecyl-1-yl)acetyl]-(D)-phenylalanyl-(L)-cysteyl-(L)-tyrosinyl-(D)-tryptophanyl-(L)-lysine-(L)-threonyl-(L)-cysteyl-(L)-threonine-acyl(2-7)disulfide ([(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], which can be used as a targeting agent. The chemical structure of DOTATATE is shown below.

[0009]

[0010] An emerging field is the use of chelated (conjugated) bioactive peptides for labeling with radioactive metals in various areas of nuclear oncology diagnosis and treatment. In recent years, several reports have documented the use of radiolabeled somatostatin analogs for targeted radiotherapy. 68 Ga-DOTATATE((Dedden SA, et al.; J Nucl Med; 2016 vol.57no.6 872-878), 68Ga-DOTATOC (Nicolas, GP, et al.; JNucl Med; 2018vol.59no.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 for NET visualization, while 177 Lu-DOTATATE has been used in radionuclide therapy (Strosberg, J. et al.; N Engl J Med 2017; 376:125-135). However, further peptide-based compounds with practical applications in diagnostic imaging techniques such as PET are needed.

[0011] Copper-64 ( 64 Copper-64 (Cu) is a positron-emitting radionuclide, making it ideal for use as a diagnostic agent in positron emission tomography (PET). Its 12.7-hour half-life allows for post-production processing, labeling, and shipping, and its average positron energy of 0.28 MeV provides high-resolution imaging. Importantly, 64 Ni(p,n) 64 The large cross-section of the Cu reaction allows for commercial-scale production. The radioisotope copper-64 (Cu-64) for PET has been radiolabeled as a chelate peptide conjugate, DOTATATE, for diagnostic imaging of neuroendocrine tumors in humans.

[0012] The complete chemical structure of the Cu-DOTATATE complex has not been determined by X-ray crystallography, but the Cu-DOTA complex has been structurally determined by X-ray crystallography. In its crystalline form, the Cu-DOTA complex has been shown to be 6-coordinated with 4 amino nitrogen atoms and 2 carboxylic acid ester oxygen atoms, as shown below.

[0013]

[0014] The two carboxylic acid groups are left-free and do not coordinate with copper ions. Therefore, it is expected that the linkage of the peptide via one of the carboxylic acids to form a linking amide bond will not alter the coordination of copper with the DOTATATE peptide.

[0015] 64 Cu-DOTATATE binds to the somatostatin receptor, exhibiting the highest affinity for the type 2 receptor (SSTR2). It binds to cells expressing the somatostatin receptor, including malignant neuroendocrine cells that overexpress the SSTR2 receptor. 64Cu is a positron-emitting (β) + The radioactive nuclide emits an emission yield that allows for positron emission tomography (PET) imaging. When 64 When Cu's imaging capabilities are combined with DOTATATE's receptor-targeting capabilities, the result is... 64 Cu DOTATATE is a radiopharmaceutical that can image neuroendocrine tumors (NETs) that express somatostatin receptors.

[0016] now, 64 Cu-DOTATATE is prepared on-site for use in a very small number of patients with low total radioactivity. However, an unmet need remains regarding the provision of a method for preparing high-purity Cu-DOTATATE. 64 Improved process for Cu-DOTATATE and expansion 64 The radiolabeling of Cu-DOTATATE is produced while maintaining sufficient stability to transport the drug product to patients in hospitals. Summary of the Invention

[0017] This disclosure addresses the aforementioned needs and relates to a method for providing copper-labeled pharmaceutical products capable of supplying commercial quantities. 64 Useful methods for Cu-DOTATATE.

[0018] The purpose of this invention is to demonstrate and confirm the following finding: labeling copper at lower temperatures (≤30°C) has the advantage of improving the purity of pharmaceutical products, because many other common metallic impurities are actually labeled with chelates (such as DOTATATE) much more slowly than copper. Previous research on copper radioisotopes (i.e....) 64 Cu、 67 Studies on the radiolabeling of Cu are typically conducted at high temperatures (e.g., 40°C to 95°C). The high temperature (elevated temperature) is used to accelerate the labeling process and ensure maximum radiolabeling of the copper chelate. Several literature reviews have shown that sufficient labeling can be obtained by performing the labeling at room temperature. This disclosure teaches that a faster copper labeling kinetic can be used to obtain a purer product compared to the slower labeling kinetics of other metals.

[0019] This article provides: 64 Preparation of Cu-DOTATATE, monitoring of final formulation parameters and their effects 64 Experimental design (DOE) to study the effect of Cu-DOTATATE stability, scaled up for the preparation of 500mCi-2000mCi. 64 Experiments with Cu-DOTATATE 64The stability of Cu-DOTATATE in its final formulation, optimization of the amount of DOTATATE used relative to the total activity of radiolabeling, and 64 The specific activity of Cu chloride solution 64 The effect of Cu-DOTATATE purity.

[0020] For example, this document provides a method for radiolabeling DOTATATE, comprising the step of reacting copper-64 with a buffer solution containing DOTATATE, wherein the reaction occurs at a temperature less than or equal to 30°C for less than 15 minutes, and the molar ratio of DOTATATE to copper-64 in the reaction solution is from about 110:1 to about 90:1.

[0021] This article further provides preparation of... 64 A method for preparing a pharmaceutical product of Cu-DOTATATE, wherein the pharmaceutical product is prepared by the following steps: (i) radiolabeling DOTATATE with copper-64 at a concentration of about 0.6 μg / mL (μg DOTATATE / mCi copper-64), and the radionuclide purity of copper-64 in the pharmaceutical product is about 99%.

[0022] This article also provides a pharmaceutical product containing 64Cu-DOTATATE for positron emission tomography, wherein... 64 Cu-DOTATATE is stored in a container containing 148 MBq. 64 The single-dose vial of Cu-DOTATATE contains a drug product with a radioactivity concentration of approximately 5-15 mCi / mL, and the drug product has a radiochemical purity of ≥96% after dilution. Attached Figure Description

[0023] The foregoing features of the embodiments will be more readily understood with reference to the following detailed description of the invention and the accompanying drawings, wherein:

[0024] Figure 1(A) shows a general radiolabeling and preparation scheme. Figure 1(B) shows the general radiolabeling and preparation scheme of the present invention.

[0025] Figure 2 Representative HPLC chromatograms of DOTATATE and gentianic acid standard solutions are given.

[0026] Figure 3 A representative HPLC chromatogram of the crude Cu-DOTATATE reaction mixture is given after mixing equimolar amounts of DOTATATE and Cu for 5 minutes at room temperature.

[0027] Figure 4Give the recovery rate of DOTATATE in the fractionated loading solution (total 12 mL) and the final 50% EtOH eluent at a flow rate of 12 mL / min.

[0028] Figure 5 Give the recovery rate of DOTATATE in the fractionated loading solution (total 18 mL) and the final 50% EtOH eluent at a flow rate of 18 mL / min. Invention Details

[0030] 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 comprehensive and complete, and will fully convey the scope of the subject matter of this disclosure to those skilled in the art. All publications, patents, and patent applications cited herein, whether mentioned above or below, are incorporated herein by reference in their entirety.

[0031] A. Definition

[0032] Unless otherwise defined, all terms and phrases used herein include their meanings as they have been acquired in the art, unless the context in which the term or phrase is used clearly indicates or obviously indicates the opposite meaning. Although any methods and materials similar to or equivalent to those described herein may be used in the practice or testing of the invention, specific methods and materials are described hereafter.

[0033] Unless otherwise stated, the use of a single numerical value is referred to as an approximation, as if the value were preceded by the word "about" or "approximately". Similarly, unless explicitly stated otherwise, numerical values ​​within the various ranges specified herein are indicated as approximations, as if the minimum and maximum values ​​within the range were preceded by "about" or "approximately". In this way, variations above and below the range can be used to obtain results substantially the same as those within the range. As used herein, when referring to numerical values, the terms "about" and "approximately" should have the simple and general meaning that is most closely related to the disclosed subject matter or to the range or element in question. The amount by which a numerical value is broadened depends on many factors. Some factors that may be considered include the criticality of the element and / or the impact of a variation in a given amount on the performance of the claimed subject matter, as well as other considerations known to those skilled in the art. The use of different significant figures for different numerical values ​​as used herein does not imply any limitation on how the terms "about" or "approximately" broaden a particular numerical value or range. Therefore, in general, "about" or "approximately" broadens the numerical value. Furthermore, the disclosure of the range is intended as a continuous range, including every value between the minimum and maximum, plus the broadening effect provided by the use of the terms "about" or "approximately". Therefore, the description of the range of values ​​in this document is intended only as a shorthand method of individually referring to each individual value falling within that range, and each individual value is incorporated into the specification as if it were described individually in this document.

[0034] The term "drug product" or " 64 "Cu-DOTATATE injection" can be used interchangeably in this article, referring to the final formulation used as a radiodiagnostic agent. 64 Cu-DOTATATE.

[0035] "Optional" or "optionally" means that the element, component, or environment described below may or may not occur, and therefore the description includes instances where the element, component, or environment occurs and instances where it does not occur.

[0036] The terms “subject” or “patient” are used interchangeably in this document and refer to humans or other mammals.

[0037] B. Introduction

[0038] This disclosure relates to the use of 64 Improved radiolabeling and formulations for large-scale preparation of Cu-DOTATATE; Figure 1(A).

[0039] Positron emission tomography (PET) offers high-resolution imaging capabilities that can be used in oncology to help clinicians better understand a patient's disease status and monitor treatment effectiveness, leading to more effective and personalized care. One such PET agent is... 64 Cu-DOTATATE, which targets and images neuroendocrine tumors (NETs) that overexpress somatostatin receptor subtype 2 (SSTR2), can help identify patients who may benefit from receptor-targeted therapy. Imaging capabilities are provided by... 64 Cu is provided. 64 Cu is a radioactive nuclide that emits positrons and can be used for PET imaging. 1 / 2 =12.7h, β + avg =0.28 MeV, I = 17.6% [representing intensity (I), sometimes reported as branching ratio (BR)], while the target portion of the molecule is a modified version of octreotate (DOTA- D -Phe-Cys-Tyr- D -Trp-Lys-Thr-Cys-Thr (a disulfide-cyclized Cys2-Cys7), a cyclic peptide that mimics the natural SSTR2 ligand somatostatin. These two functions are linked by DOTA, a bifunctional chelator that captures... 64 While Cu is present, it also binds to the N-terminus of the peptide (forming DOTATATE). 64 The structure of Cu-DOTATATE (Cu 64DOTATATE) is as follows.

[0040]

[0041] 64 Structure of Cu-DOTATATE (Copper Cu 64DOTATATE)

[0042] Dotatate and radioactive isotopes 64 Radiolabeling of Cu was first performed decades ago. In past studies, small batches... 64 Cu-DOTATATE is prepared only at sites with low total radioactivity for very limited use and a small number of patients. Recently, radiolabeling has been improved to achieve higher levels of radioactivity, allowing for scale-up to commercial production. The final purified product is prepared at much higher initial total radioactivity levels, and improved formulations and purification methods prevent radioactive degradation.

[0043] There is a need to scale up the formulation of radiolabeled and high-volume drugs in order to enable the distribution of said drugs throughout the country. Scaling up brings new problems and challenges, as well as new discoveries and solutions for achieving high-volume drug production. This disclosure (i) explains improvements and changes to past research and results, and (ii) teaches the research conducted and how to scale up to highly active, high-volume production.

[0044] The commonly used radiolabeling and preparation schemes are shown in Figure 1(B).

[0045] Specifically, this public teaching 64 A significant amplification of the total radioactivity of Cu. 64 Cu can be radiolabeled and purified for use 64 Cu-DOTATATE injectable drug product.

[0046] Previous research is 64 The radioactivity of Cu is at low to moderate levels (mCi). This disclosure amplifies the total radioactivity during radiolabeling to >5400 mCi. The challenge lies in achieving radiolabeling without radioactive decomposition and due to dotatate and other factors. 64 The radiolabeled product degrades due to competition from metals other than Cu. It must then be rapidly purified and immediately diluted in a stable solution to prevent degradation caused by radioactive decomposition, thereby maintaining the required high radiochemical purity (RCP).

[0047] Up to 48 hours after labeling, the purified 64 The stability of Cu-DOTATATE in its final formulation (5% ethanol in 45 mg / mL sodium ascorbate) was evaluated and showed... 64 Cu was not significantly degraded or lost from the complex.

[0048] In order to prepare 64 Cu-DOTATATE, in dilute HCl 64 CuCl2 was reacted with DOTATATE in sodium acetate buffer containing gentian acid at a ratio of 2 μg DOTATATE / mCi. The reaction mixture was cultured and then purified into sodium ascorbate (NaOAsc) buffer. The resulting... 64 The Cu-DOTATE solution was aseptically filtered and used as the final formulation. The development work disclosed herein focuses on improving the production design space and scaling up the radiolabeling reaction to prepare ≥2Ci. 64 Cu-DOTATATE. Radiolabeling has been demonstrated to occur within 5 minutes, even at 15°C. The purified product has a concentration up to 10,000 mCi. 64The procedure was performed under Cu-DOTATATE. The purified product was obtained using 50% ethanol in water (previous literature indicated that only pure ethanol was used). In fact, using 50% ethanol in water increased the yield of the purified product compared to using 100% ethanol.

[0049] The purified drug product (2 mL) was immediately diluted to a large volume (>20 mL; but typically >100 mL for 2000–10000 mCi products) to prevent degradation (radioactive decomposition) and to maintain the required RCP >95%. Previous literature diluted the purified product to <20 mL.

[0050] The final drug product was stabilized for up to 48 hours after purification using 28-122 mg / mL sodium ascorbate and 1-5% ethanol, with an RCP > 95%. Previous 48-hour RCP stabilization was achieved using only 45 mg / mL sodium ascorbate / 5% ethanol.

[0051] Initial labeling (radioactive labeling step) can be achieved in the presence of sodium ascorbate.

[0052] Surprisingly, it was found that when DOTATATE was labeled at lower temperatures (i.e., ≤30°C), the chelation of DOTATATE with copper occurred faster than with other metals. This phenomenon could be used to reduce the amount of metal impurities present in the final pharmaceutical product.

[0053] C. 64 Preparation of Cu-DOTATATE Bulk Solution

[0054] i. Ligands

[0055] In one embodiment, the ligand is DOTATATE; 1,4,7,10-tetraazacyclododecane-1,4,7,10-tretraacetic 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.

[0056] In one embodiment, the ligand is added to the reaction mixture in amounts of about 1 μg to about 6000 μg, about 50 μg to 5000 μg, about 100 μg to 4500 μg, about 200 μg to 4000 μg, about 300 μg to 3000 μg, about 400 μg to 2000 μg, and about 500 μg to 1000 μg. In another embodiment, the ligand is added to the reaction mixture in amounts of: 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. Approximately 1900 μg, approximately 2000 μg, approximately 2100 μg, approximately 2200 μg, approximately 2300 μg, approximately 2400 μg, approximately 2500 μg, approximately 2600 μg, approximately 2700 μg, approximately 2800 μg, approximately 2900 μg, approximately 3000 μg, approximately 3100 μg, approximately 3200 μg, approximately 3300 μg, approximately 4400 μg, approximately 4500 μg, approximately 5000 μg, approximately 5500 μg, or approximately 6000 μg. In another embodiment, the ligand is added to the reaction mixture in amounts of 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, 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, and less than Approximately 1900 μg, less than approximately 2000 μg, less than approximately 2100 μg, approximately 2200 μg, less than approximately 2300 μg, less than approximately 2400 μg, less than approximately 2500 μg, less than approximately 2600 μg, less than approximately 2700 μg, less than approximately 2800 μg, less than approximately 2900 μg, less than approximately 3000 μg, less than approximately 3100 μg, less than approximately 3200 μg, less than approximately 3300 μg, less than approximately 4400 μg, less than approximately 4500 μg, less than approximately 5000 μg, less than approximately 5500 μg, or less than approximately 6000 μg.

[0057] In another embodiment, the ligand is used in amounts 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 another embodiment, the ligand is used in amounts of approximately 0.1 ug / mCi, approximately 0.25 ug / mCi, approximately 0.4 ug / mCi, approximately 0.5 ug / mCi, approximately 0.6 ug / mCi, approximately 0.75 ug / mCi, approximately 0.8 ug / mCi, approximately 1 ug / mCi, approximately 1.25 ug / mCi, approximately 1.5 ug / mCi, approximately 1.75 ug / mCi, approximately 2 ug / mCi, approximately 2.5 ug / mCi, approximately 3 ug / mCi, approximately 3.5 ug / mCi, or approximately 4 ug / mCi.

[0058] 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.

[0059] In another embodiment, the total labeled ligands are about 200 μg to about 6000 μg, about 500 μg to about 5000 μg, about 1000 μg to 4000 μg, about 1500 μg to 3000 μg, about 2000 μg to 25000 μg, about 2000 μg to about 4000 μg, or about 3000 μg to about 4000 μg. In another embodiment, the total labeled ligands are approximately 200 μg, approximately 300 μg, approximately 400 μg, approximately 500 μg, approximately 600 μg, approximately 700 μg, approximately 800 μg, approximately 900 μg, approximately 1000 μg, approximately 1100 μg, approximately 1200 μg, approximately 1300 μg, approximately 1400 μg, approximately 1500 μg, approximately 1600 μg, approximately 1700 μg, approximately 1800 μg, and approximately 1900 μg. Approximately 2000 μg, approximately 2100 μg, approximately 2200 μg, approximately 2300 μg, approximately 2400 μg, approximately 2500 μg, approximately 2600 μg, approximately 2700 μg, approximately 2800 μg, approximately 2900 μg, approximately 3000 μg, approximately 3100 μg, approximately 3200 μg, approximately 3300 μg, approximately 4000 μg, approximately 4500 μg, approximately 5000 μg, approximately 5500 μg, or approximately 6000 μg. In another embodiment, the total labeled ligands are 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.

[0060] ii. Radionuclides

[0061] In another embodiment, the radionuclides are 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, technetium-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, thallium-201, etc.

[0062] In another implementation scheme, 64 CuCl2 was added to the reaction mixture in the following amounts (as...). 64Cu source: approximately 100 mCi to approximately 5000 mCi, approximately 200 mCi to approximately 4000 mCi, approximately 300 mCi to approximately 3500 mCi, approximately 400 mCi to 3000 mCi, approximately 500 mCi to 2500 mCi, or up to approximately 10000 mCi. In one embodiment, 64 CuCl2 was added to the reaction mixture in the following amounts (as...). 64 Cu source): approximately 100 mCi, approximately 200 mCi, approximately 300 mCi, approximately 400 mCi, approximately 500 mCi, approximately 600 mCi, approximately 700 mCi, approximately 800 mCi, approximately 900 mCi, approximately 1000 mCi, approximately 1500 mCi, approximately 2000 mCi, approximately 2500 mCi, approximately 3000 mCi, approximately 3 500 mCi, approximately 4000 mCi, approximately 4500 mCi, approximately 5000 mCi, approximately 5500 mCi, approximately 6000 mCi, approximately 6500 mCi, approximately 7000 mCi, approximately 7500 mCi, approximately 8000 mCi, approximately 8500 mCi, approximately 9000 mCi, approximately 9500 mCi, or approximately 10000 mCi. In another embodiment, 64 CuCl2 was added to the reaction mixture in the following amounts (as...). 64 Cu source): less than approximately 100 mCi, less than approximately 200 mCi, less than approximately 300 mCi, less than approximately 400 mCi, less than approximately 500 mCi, less than approximately 600 mCi, less than approximately 700 mCi, less than approximately 800 mCi, less than approximately 900 mCi, less than approximately 1000 mCi, less than approximately 1500 mCi, less than approximately 2000 mCi, less than approximately 2500 mCi, less than approximately 3000 mCi, less than Approximately 3500 mCi, less than approximately 4000 mCi, less than approximately 4500 mCi, less than approximately 5000 mCi, less than approximately 5500 mCi, less than approximately 6000 mCi, less than approximately 6500 mCi, less than approximately 7000 mCi, less than approximately 7500 mCi, less than approximately 8000 mCi, less than approximately 8500 mCi, less than approximately 9000 mCi, less than approximately 9500 mCi, or less than approximately 10000 mCi.

[0063] In one embodiment, the radionuclide is added to the reaction mixture in amounts 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.

[0064] In another implementation scheme,64 Cu is added to the reaction mixture in the following amounts: 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, 5 μg, about 6 μg, about 7 μg, about 8 μg, about 9 μg, or 10 μg.

[0065] iii. Buffer solution

[0066] In one embodiment, the buffer solution used to prepare the bulk solution of the pharmaceutical product is a sodium acetate buffer, a sodium acetate / gentianic acid buffer, a sodium ascorbate buffer, a sodium ascorbate / ethanol buffer, an ammonium acetate buffer, an ammonium acetate / gentianic acid buffer, an ammonium ascorbate buffer, or an ammonium ascorbate / ethanol buffer, or any other suitable buffer.

[0067] In one embodiment, the concentration of the buffer is about 0.1M, about 0.2M, about 0.3M, about 0.4M, about 0.5M, about 0.6M, about 0.7M, about 0.8M, about 0.9M, or about 1.0M. In 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 120 mg / mL, or about 70 mg / mL to 100 mg / mL. In another embodiment, the concentration of the buffer solution 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.

[0068] 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 100 mg / mL gentianic 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.

[0069] In one specific implementation, the buffer is a solution of 4 mg / mL gentian acid and 0.4 M sodium acetate.

[0070] In 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, about 100 mg / mL 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 having 45 mg / mL sodium ascorbate and 5% EtOH.

[0071] 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 100 mg / mL gentianic 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.

[0072] iv. Stabilizer

[0073] In one embodiment, the stabilizer is gentianic acid. In another embodiment, the stabilizer is sodium ascorbate. However, any suitable stabilizer may be used.

[0074] In another embodiment, more than one stabilizer is used. In yet another embodiment, one stabilizer, such as gentianic acid, is used during the radiolabeling process, and another stabilizer, such as sodium ascorbate, is used in the final formulated product.

[0075] 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 a specific embodiment, the stabilizer is added to the reaction mixture in an amount of about 4.0 mg.

[0076] In another embodiment, the stabilizer is added in an amount of about 1.0 g to about 9.0 g. In another embodiment, gentic acid is added in an amount of about 2.0 mg to about 8.0 mg. In another embodiment, gentic acid is added in an amount of about 3.0 mg to about 7.0 mg. In another embodiment, gentic acid is added in an amount of about 3.0 mg to about 5.0 mg. In another embodiment, gentic acid is added in an amount of about 4.0 mg to about 6.0 mg. In one specific embodiment, gentic acid is added to the reaction mixture in an amount of about 4.0 mg.

[0077] 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.

[0078] v. Conditions for radioactive labeling

[0079] In one embodiment, radiolabeling is performed at radionuclides ranging from 500 mCi to 15000 mCi. The radioactive concentrations during labeling are ≥250 mCi / mL, ≥300 μg / mL, ≥333 mCi / mL, ≥350 mCi / mL, ≥400 mCi / mL, ≥421 mCi / mL, or ≥460 mCi / mL. The total labeled ligands are 1000-4000 μg or have a concentration ≥333 μg / mL.

[0080] In one embodiment, radiolabeling is performed with a radionuclide at a concentration of 500 mCi to 10000 mCi. The radioactive concentration during labeling is ≥333 mCi / mL. The total labeled ligand is 1000-4000 μg or at a concentration ≥333 μg / mL.

[0081] In another embodiment, radiolabeling is performed at a radionuclide concentration of 500 mCi to 2500 mCi. The radioactive concentration during labeling is ≥250 mCi / mL, ≥300 μg / mL, ≤333 mCi / mL, ≥350 mCi / mL, ≥400 mCi / mL, ≥421 mCi / mL, or ≥460 mCi / mL. The total labeled ligand is 1000-4000 μg or has a concentration ≥333 μg / mL.

[0082] In one embodiment, the radioactive label is in the range of 500 mCi to 15000 mCi. 64 The process is carried out under Cu. The radioactive concentration during labeling is ≥250 mCi / mL, ≥300 mCi / mL, ≥333 mCi / mL, ≥350 μg / mL, ≥400 mCi / mL, ≥421 mCi / mL, or ≥460 mCi / mK. The total amount of labeled DOTATATE is 1000-4000 μg or the concentration is ≥333 μg / mL.

[0083] In one embodiment, the radioactive label is in the range of 500 mCi to 10000 mCi. 64 The procedure is performed under Cu. The radioactive concentration during radiolabeling is ≥333 mCi / mL. The total amount of labeled DOTATATE is 1000-4000 ug or the concentration is ≥333 ug / mL.

[0084] In another embodiment, the radioactive label is between 500 mCi and 2500 mCi. 64 The process is carried out under Cu. The radioactive concentration during labeling 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 amount of labeled DOTATATE is 1000–4000 μg or the concentration is ≥333 μg / mL.

[0085] 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 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.

[0086] Bioconjugated chelates (e.g., DOTA-TATE) are typically complexed at temperatures of 50–95 °C to ensure high radiochemical and radiolabeling yields. However, this disclosure teaches that copper can be labeled at lower temperatures (i.e., room temperature or lower). 64 The purity of Cu-DOTATATE was improved because copper (2+) ions were labeled onto DOTATATE more quickly than other common metal impurities.

[0087] 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 another 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 yet another embodiment, the temperature of the reaction mixture is ambient temperature.

[0088] In another embodiment, the temperature of the reaction mixture is below or equal to 50°C, below 50°C, below or equal to 45°C, below 45°C, below or equal to 40°C, below 40°C, below or equal to 35°C, below 35°C, below or equal to 30°C, below 30°C, below or equal to 25°C, below 25°C, below or equal to 20°C, below 20°C, below or equal to 15°C, below 15°C, below or equal to 10°C, or below 10°C.

[0089] In one specific embodiment, the molar ratio of the ligand to the radionuclide in the reaction mixture is about 125:1, about 120:1, about 115:1, about 110:1, about 105:1, about 100:1, about 95:1, about 90:1, about 85:1, about 80:1, about 75:1, about 70:1, about 65:1, about 60:1, about 55:1, about 50:1, about 45:1, about 40:1, about 35:1, about 30:1, about 25:1, about 20:1, about 15:1, about 10:1, about 5:1, about 4:1, about 3:1, about 2:1, about 2.5:1, or 1:1. In another embodiment, the molar ratio of the ligand to the radionuclide in the reaction mixture is about 125:1 to about 75:1. In another embodiment, the molar ratio of ligand to radionuclide in the reaction mixture is from about 105:1 to about 95:1. In another embodiment, the molar ratio of ligand to radionuclide in the reaction mixture is from about 110:1 to about 90:1. In 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.

[0090] In one specific implementation, DOTATATE and 64 The molar ratio of Cu is about 125:1, about 120:1, about 115:1, about 110:1, about 105:1, about 100:1, about 95:1, about 90:1, about 85:1, about 80:1, about 75:1, about 70:1, about 65:1, about 60:1, about 55:1, about 50:1, about 45:1, about 40:1, about 35:1, about 30:1, about 25:1, about 20:1, about 15:1, about 10:1, about 5:1, about 4:1, about 3:1, about 2:1, about 2.5:1, or about 1:1. In another embodiment, DOTATATE in the reaction mixture is... 64 The molar ratio of Cu is approximately 105:1 to approximately 95:1. In another embodiment, DOTATATE in the reaction mixture... 64 The molar ratio of Cu is from about 102:1 to about 99:1. In another embodiment, DOTATATE in the reaction mixture is... 64The molar ratio of 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.

[0091] In one embodiment, the ratio of the mass (μg) of the ligand to the radioactivity (mCi) of the radionuclide is about 5:1, about 4:1, about 3:1, about 2:1, or about 1:1. In another embodiment, for each reaction, the ratio of the mass (μg) of the ligand to the radioactivity of the radionuclide (mCi) concentration is about 1.0 μg / mCi, about 0.9 μg / mCi, about 0.8 μg / mCi, about 0.7 μg / mCi, about 0.6 μg / mCi, about 0.5 μg / mCi, about 0.4 μg / mCi, about 0.3 μg / mCi, about 0.2 μg / mCi, or 0.1 μg / mCi. In yet another embodiment, for each reaction, the ratio of the mass (μg) of the ligand to the radioactivity (mCi) of the radionuclide is about 0.6 μg / mCi.

[0092] In one implementation, the ligand mass (μg): 64 The radioactivity ratio of Cu(mCi) is approximately 5:1, approximately 4:1, approximately 3:1, approximately 2:1, or approximately 1:1. In another embodiment, for each reaction, the ratio of the mass (μg) of the ligand to the radioactivity (mCi) of the radionuclide is approximately 1.0 μg / mCi, approximately 0.9 μg / mCi, approximately 0.8 μg / mCi, approximately 0.7 μg / mCi, approximately 0.6 μg / mCi, approximately 0.5 μg / mCi, approximately 0.4 μg / mCi, approximately 0.3 μg / mCi, approximately 0.2 μg / mCi, or 0.1 μg / mCi. In another embodiment, for each reaction, the ratio of the mass (μg) of the ligand to the radioactivity (mCi) of the radionuclide is approximately 0.6 μg / mCi.

[0093] In one implementation, the mass (μg) of DOTATATE is related to 64 The radioactivity ratio of Cu(mCi) is approximately 5:1, approximately 4:1, approximately 3:1, approximately 2:1, or approximately 1:1. In another embodiment, for each reaction, the mass (μg) of DOTATATE is... 64The radioactivity ratios of Cu(mCi) concentrations are approximately 1.0 μg / mCi, approximately 0.9 μg / mCi, approximately 0.8 μg / mCi, approximately 0.7 μg / mCi, approximately 0.6 μg / mCi, approximately 0.5 μg / mCi, approximately 0.4 μg / mCi, approximately 0.3 μg / mCi, approximately 0.2 μg / mCi, or approximately 0.1 μg / mCi. In another embodiment, for each reaction, the mass (μg) of DOTATATE is... 64 The radioactivity ratio of Cu(mCi) concentration is approximately 0.6 μg / mCi.

[0094] In one embodiment, the radioactivity of the bulk solution of the drug substance is approximately 1 mCi to approximately 10,000 mCi, approximately 1 mCi to approximately 9,900 mCi, approximately 1 mCi to approximately 9,800 mCi, approximately 1 mCi to approximately 9,700 mCi, approximately 1 mCi to approximately 9,600 mCi, approximately 1 mCi to approximately 9,500 mCi, approximately 1 mCi to approximately 9,400 mCi, approximately 1 mCi to approximately 9,300 mCi, approximately 1 mCi to approximately 9,200 mCi, approximately 1 mCi to approximately 9,100 mCi, approximately 1 mCi to approximately 9,000 mCi, approximately 1 mCi to approximately 8,900 mCi, approximately 1 mCi to approximately 8,800 mCi, and approximately 1 mCi to approximately 8,700 mCi. Approximately 1 mCi to approximately 8,600 mCi, approximately 1 mCi to approximately 8,500 mCi, approximately 1 mCi to approximately 8,400 mCi, approximately 1 mCi to approximately 8,300 mCi, approximately 1 mCi to approximately 8,200 mCi, approximately 1 mCi to approximately 8,100 mCi, approximately 1 mCi to approximately 8,000 mCi, approximately 1 mCi to approximately 7,900 mCi, approximately 1 mCi to approximately 7,800 mCi, approximately 1 mCi to approximately 7,700 mCi, approximately 1 mCi to approximately 7,600 mCi, approximately 1 mCi to approximately 7,500 mCi, approximately 1 mCi to approximately 7,400 mCi, approximately 1 mCi to approximately 7,300 mCi, approximately 1 mCi to approximately 7,200 mCi, approximately 1 m Ci to approximately 7,100 mCi, approximately 1 mCi to approximately 7,000 mCi, approximately 1 mCi to approximately 6,900 mCi, approximately 1 mCi to approximately 6,800 mCi, approximately 1 mCi to approximately 6,700 mCi, approximately 1 mCi to approximately 6,600 mCi, approximately 1 mCi to approximately 6,500 mCi, approximately 1 mCi to approximately 6,400 mCi, approximately 1 mCi to approximately 6,300 mCi, approximately 1 mCi to approximately 6,200 mCi, approximately 1 mCi to approximately 6,100 mCi, approximately 1 mCi to approximately 6,000 mCi, approximately 1 mCi to approximately 5,900 mCi, approximately 1 mCi to approximately 5,800 mCi, approximately 1 mCi to approximately 5,700 mCi, approximately 1 mCi From about 5,600 mCi, about 1 mCi to 5,500 mCi, about 1 mCi to about 5,400 mCi, about 1 mCi to about 5,300 mCi, about 1 mCi to about 5,200 mCi, about 1 mCi to about 5,100 mCi, about 1 mCi to about 5,000 mCi, about 1 mCi to about 4,900 mCi, about 1 mCi to about 4,800 mCi, about 1 mCi to about 4,700 mCi, about 1 mCi to about 4,600 mCi, about 1 mCi to about 4,500 mCi, about 1 mCi to about 4,400 mCi, about 1 mCi to about 4,300 mCi, about 1 mCi to about 4,200 mCi, about 1 mCi to about 4,100 mCi, about 1 mCi to about 4,000 mCi, about 1 mCi to about 3,900 mCi, about 1 mCi to about 3,800 mCi, about 1 mCi to about 3,700 mCi, about 1 mCi to about 3,600 mCi, about 1 mCi to about 3,500 mCi, about 1 mCi to about 3,400 mCi, about 1 mCi to about 3,300 mCi, about 1 mCi to about 3,200 mCi, about 1 mCi to about 3,100 mCi, about 1 mCi to about 3,000 mCi, about 10 mCi to about 2,900 mCi, about 20 mCi to about 2,800 mCi, about 30 mCi to about 2,700 mCi 00 mCi, about 40 mCi to 2,600 mCi, about 50 mCi to 2,500 mCi, about 60 mCi to 2,400 mCi, about 70 mCi to 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.

[0095] In another embodiment, the radioactivity of the bulk solution of the pharmaceutical substance is approximately 1 mCi, approximately 20 mCi, approximately 40 mCi, approximately 60 mCi, approximately 80 mCi, approximately 100 mCi, approximately 120 mCi, approximately 140 mCi, 160 mCi, approximately 200 mCi, approximately 220 mCi, approximately 240 mCi, approximately 260 mCi, approximately 280 mCi, approximately 300 mCi, approximately 320 mCi, approximately 340 mCi, approximately 360 mCi, approximately 380 mCi, approximately 400 mCi, approximately 420 mCi, approximately 440 mCi, approximately 460 mCi, approximately 480 mCi, approximately 500 mCi, approximately 550 mCi, approximately 600 mCi, approximately 650 mCi, and approximately 700 mCi. Approximately 750 mCi, approximately 800 mCi, approximately 850 mCi, approximately 900 mCi, approximately 950 mCi, approximately 1,000 mCi, approximately 1,100 mCi, approximately 1,200 mCi, approximately 1,300 mCi, approximately 1,400 mCi, approximately 1,500 mCi, approximately 1,600 mCi, approximately 1,700 mCi, approximately 1, 800mCi, approximately 1,900mCi, approximately 2,000mCi, approximately 2,100mCi, approximately 2,200mCi, approximately 2,300mCi, approximately 2,400mCi, approximately 2,500mCi, approximately 2,600mCi, approximately 2,700mCi, approximately 2,800mCi, approximately 2,900mCi, approximately 3,000mCi Ci, approximately 3,100 mCi, approximately 3,200 mCi, approximately 3,300 mCi, approximately 3,400 mCi, approximately 3,500 mCi, approximately 3,600 mCi, approximately 3,700 mCi, approximately 3,800 mCi, approximately 3,900 mCi, approximately 4,000 mCi, approximately 4,100 mCi, approximately 4200 mCi, approximately 4,300 mCi, approximately 4,400 mCi, approximately 4,500 mCi, approximately 4,600 mCi, approximately 4,700 mCi, approximately 4,800 mCi, approximately 4,900 mCi, approximately 5,000 mCi, approximately 5,100 mCi, approximately 5,200 mCi, approximately 5,300 mCi, approximately 5,400 mCi, approximately 5,500 mCi 0 mCi, approximately 5,600 mCi, approximately 5,700 mCi, approximately 5,800 mCi, approximately 5,900 mCi, approximately 6,000 mCi, approximately 6,100 mCi, approximately 6,200 mCi, approximately 6,300 mCi, approximately 64,000 mCi, approximately 6,500 mCi, approximately 6,600 mCi, approximately 6,700 mCi Approximately 6,800 mCi, approximately 6,900 mCi, approximately 7,000 mCi, approximately 7,100 mCi, approximately 7,200 mCi, approximately 7,300 mCi, approximately 7,400 mCi, approximately 7,500 mCi, approximately 7,600 mCi, approximately 7,700 mCi, approximately 7,800 mCi, approximately 7,900 mCi, approximately 8,000mCi, approximately 8,100mCi, approximately 8,200mCi, approximately 8,300mCi, approximately 8,400mCi, approximately 8,500mCi, approximately 8,600mCi, approximately 8,700mCi, approximately 8,800mCi, approximately 8,900mCi, approximately 9,000mCi, approximately 9,100mCi, approximately 9,200mCi, approximately 9,300mCi, approximately 9,400mCi, approximately 9,500mCi, approximately 9,600mCi, approximately 9,700mCi, approximately 9,800mCi, approximately 9,900mCi, or approximately 10,000mCi. In one specific implementation, the radioactivity of the bulk solution of the pharmaceutical substance is approximately 100 mCi, approximately 500 mCi, approximately 1,000 mCi, approximately 2,000 mCi, approximately 3,000 mCi, approximately 4,000 mCi, approximately 5,000 mCi, approximately 6,000 mCi, approximately 7,000 mCi, approximately 8,000 mCi, approximately 9,000 mCi, or approximately 10,000 mCi.

[0096] 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 approximately 0.1 mL to approximately 10 mL, approximately 0.2 mL to approximately 9 mL, approximately 0.3 mL to approximately 8 mL, approximately 0.4 mL to approximately 7 mL, approximately 0.5 mL to approximately 6 mL, approximately 1 mL to approximately 5 mL, and approximately 2 mL to approximately 4 mL. In another embodiment, 64 The volume of Cu solution is approximately 0.1 mL, 0.2 mL, 0.3 mL, 0.4 mL, 0.5 mL, 1 mL, 2 mL, 3 mL, 4 mL, 5 mL, 6 mL, 7 mL, 8 mL, 9 mL, or 10 mL.

[0097] In 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.

[0098] In another embodiment, the response time is approximately 1 minute, approximately 2 minutes, approximately 3 minutes, approximately 4 minutes, approximately 5 minutes, approximately 6 minutes, approximately 7 minutes, approximately 8 minutes, approximately 9 minutes, approximately 10 minutes, approximately 15 minutes, approximately 20 minutes, approximately 25 minutes, approximately 30 minutes, approximately 45 minutes, approximately 1 hour, approximately 2 hours, approximately 3 hours, approximately 4 hours, approximately 5 hours, approximately 6 hours, approximately 7 hours, approximately 8 hours, approximately 9 hours, or approximately 10 hours; the response time is approximately 1 minute to approximately 24 hours, approximately 1 minute to approximately 18 hours, approximately 1 minute to approximately 12 hours, or approximately 1 minute to approximately 6 hours. In another embodiment, the response time is approximately 1 minute to approximately 60 minutes, approximately 2 minutes to approximately 45 minutes, or approximately 5 minutes to approximately 30 minutes.

[0099] In another embodiment, the concentration of the antiradiolytic agent in the final formulation is 29-122 mg / mL + 1-5% ethanol.

[0100] In another embodiment, the amount of non-radioactive copper added to the reaction mixture is 0-30 μg / mL (ppm). In another embodiment, the amount of non-radioactive copper added to the reaction mixture is 0.1-30 μg / mL (ppm).

[0101] D. 64 Purification of Cu-DOTATATE bulk solution

[0102] In another embodiment, the metal ligand bulk solution is purified using C-18 Light Sep Pak or any suitable purification system / column.

[0103] In one embodiment, the elution solvent in the purification is ethanol, about 5% ethanol (95% water), about 10% ethanol (90% water), about 15% ethanol (85% water), about 20% ethanol (80% water), about 25% ethanol (75% water), about 30% ethanol (70% water), about 35% ethanol (65% water), about 40% ethanol (60% water), about 45% ethanol (55% water), about 50% ethanol (50% water), about 55% ethanol (45% water), about 60% ethanol (40% water), about 65% ethanol (35% water), 70% ethanol (30% water), about 75% ethanol (25% water), about 80% ethanol (20% water), about 85% ethanol (15% water), about 90% ethanol (10% water), about 95% ethanol (5% water), or 100% ethanol.

[0104] In 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.

[0105] In another implementation scheme, in the pharmaceutical 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%.

[0106] In another embodiment, the amount of radionuclide impurities in the pharmaceutical product is ≤1%, ≤0.9%, ≤0.8%, ≤0.7%, ≤0.6%, ≤0.5%, ≤0.4%, ≤0.3%, ≤0.2%, or ≤0.1%. In yet another embodiment, the amount of one single radionuclide impurity in the pharmaceutical product is ≤0.1%, ≤0.09%, ≤0.08%, ≤0.07%, ≤0.06%, ≤0.05%, ≤0.04%, ≤0.03%, ≤0.02%, or ≤0.01%.

[0107] In one embodiment, the radiochemical purity of the pharmaceutical 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 Cu 64DOTATATE.

[0108] In one implementation, purity is measured using high-performance liquid chromatography (HPLC) or any other acceptable or suitable technique.

[0109] In another embodiment, gentic acid is present in the pharmaceutical product in amounts of ≤50ppm, ≤40ppm, ≤30ppm, ≤20ppm, ≤10ppm, ≤5ppm or ≤1ppm.

[0110] In one embodiment, the 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 pharmaceutical product.

[0111] In another embodiment, the total impurities are present in amounts 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 pharmaceutical product.

[0112] In another embodiment, the amount of bacterial endotoxin present in the pharmaceutical product is ≤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.

[0113] F. Pharmaceutical products ( 64 Cu-DOTATATE injection)

[0114] The drug product disclosed in this article is suitable for positron emission tomography (PET) for the localization of somatostatin receptor-positive neuroendocrine tumors (NETs) in adult patients.

[0115] i. Chemical properties

[0116] The pharmaceutical products mentioned in this article contain 64 Cu-DOTATATE (copper) is a radiodiagnostic agent used in PET imaging. Chemically, 64 Cu-DOTATATE is described as copper(Cu64)-N-[(4,7,10-tricarboxymethyl-1,4,7,10-tetraazacyclododec-1-yl)acetyl]-Dphenylalanyl-L-cysteinyl-L-tyrosyl-D-tryptophanyl-L-lysyl-L-threoninyl-L-cysteinyl-L-threonine-cyclic(2-7)disulfide. With a molecular weight of 1497.2 Daltons, the following is the structural formula of one isomer:

[0117]

[0118] This drug product is a sterile, clear, colorless to yellow intravenous solution. At the calibration date and time, each 10 mL single-dose vial contains 148 MBq (4 mCi) in a 4 mL solution volume. 64 Cu-DOTATATE. In addition, each mL of solution contains 40 mg ascorbic acid, 0.05 mL anhydrous ethanol, and USP (ethanol) in sterile water for injection. The pH is adjusted with sodium hydroxide and hydrochloric acid to approximately 5.5 to 7.5.

[0119] ii. Physical properties

[0120] Tables 1 and 2 provide 64 Key radiation emission data and physical decay of Cu. 64 Cu half-life t 1 / 2 =12.7 hours, decaying through the following combination of processes: (a) 17.6% of positrons are emitted to 64Ni, which resulted in two 511 keV annihilation photon emissions (35.7%), and (b) 38.5% β decay to 64 Zn, and (c) 43.8% of electrons are captured to 64 The decay of Ni.Cu-64 also results in the emission of characteristic 1346 keV gamma rays with an intensity of approximately 0.48%.

[0121] The gamma emission spectrum of the drug product shows peaks at approximately 511 keV and approximately 1346 keV.

[0122] Table 1. Major Radiated Emission Data (>1%)

[0123]

[0124] Table 2. 64 Physical decay table of Cu

[0125] Hour Number of remaining portions Hour Number of remaining portions 0 1 18 0.374 1. 0.947 24 (1 day) 0.270 3. 0.849 36 (1.5 days) 0.140 6. 0.721 48 (2 days) 0.073 9 0.612 72 (3 days) 0.020 12 0.520 96 (4 days) 0.005

[0126] iii. External radiation

[0127] Gamma constant: 3.6 x 10⁻⁶ at 1 meter -5 mSv / hr / MBq (0.133 mrem / hr / mCi at 1 meter). Table 3 shows the lead shielding. 64 Radiation attenuation of Cu.

[0128] Table 3. Lead Shielding 64 Radiative attenuation of Cu

[0129]

[0130] In one embodiment, the pharmaceutical 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 pharmaceutical 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 pharmaceutical product is stored at a controlled room temperature of about 20°C to about 25°C.

[0131] In another embodiment, the pharmaceutical 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 pharmaceutical 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 another embodiment, the pharmaceutical product is stored at a temperature of about 50°C to about 55°C.

[0132] Radiochemical properties can be confirmed using HPLC. 64The HPLC relative retention time (RRT) of Cu-DOTATATE is related to the relative retention time of the DOTATATE standard. In one embodiment, 64 The HPLC RRT of Cu-DOTATATE is about 1 to about 2 or about 1.15 to about 1.25.

[0133] The properties of radionuclides can be confirmed using gamma emission spectroscopy. The gamma emission spectrum of the pharmaceutical product shows peaks at approximately 511 keV and approximately 1346 keV.

[0134] In one embodiment, the solution volume of the pharmaceutical 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 another embodiment, the solution volume of the pharmaceutical 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.

[0135] In another embodiment, at the calibration date and time, the pharmaceutical product is a sterile, clear, colorless to yellow solution in a single-dose vial containing 148 MBq (4 mCi) (37 MBq (1 mCi) / 1 mL). 64 Cu-DOTATATE. Sealed vials are packaged in shielded (lead) containers for radiation protection. This pharmaceutical product is shipped in Category A packaging.

[0136] In one implementation, the total radioactivity (test) of the vials is about 1.0 mCi / via to about 10 mCi / via, about 1.5 mCi / via to about 9 mCi / via, about 2.0 mCi / via to about 8 mCi / via, about 2.5 mCi / via to about 7 mCi / via, about 3.0 mCi / via to about 6 mCi / via, or about 3.6 mCi / via to about 4.4 mCi / via. In another embodiment, the total radioactivity (test) of the vials is approximately 1.0 mCi / via, approximately 1.5 mCi / via, approximately 2.0 mCi / via, approximately 2.5 mCi / via, approximately 3.0 mCi / via medium, approximately 3.5 mCi / via small, approximately 3.6 mCi / via, approximately 4.0 mCi / via body, approximately 4.4 mCi / via, approximately 4.5 mCi / via, approximately 5.0 mCi / via, approximately 5.5 mCi / via, approximately 6 mCi / via, approximately 7 mCi / via, approximately 8 mCi / via, approximately 9 mCi / via, or approximately 10 mCi / via.

[0137] In another embodiment, the radioactivity concentration of the pharmaceutical product is about 0.5 mCi / mL to about 15 mCi / mL, about 0.5 mCi / mL to about 12.5 mCi / mL, about 0.5 mCi / mL to about 10 mCi / mL, about 0.5 mCi / mL to about 7.5 mCi / mL, about 0.5 mCi / mL to about 5 mCi / mL, about 0.5 mCi / mL to about 3 mCi / mL, about 0.6 mCi / mL to about 2.5 mCi / mL, about 0.7 mCi / mL to about 2.0 mCi / mL, about 0.8 mCi / mL to about 1.5 mCi / mL, or about 0.9 mCi / mL to about 1.1 mCi / mL. In another embodiment, the radioactivity concentration of the pharmaceutical 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 pharmaceutical product is about 5-15 mCi / mL. In another embodiment, the radioactivity concentration of the pharmaceutical product is about 9-14 mCi / mL. In another embodiment, the radioactivity concentration of the pharmaceutical product is about 10-11 mCi / mL. In another embodiment, the radioactivity concentration of the pharmaceutical product is about 11-12 mCi / mL. In another embodiment, the radioactivity concentration of the pharmaceutical product is about 12-13 mCi / mL.

[0138] In another embodiment, the amount of DOTATATE and related substances present in the pharmaceutical product is ≤50ppm, about ≤40ppm, about ≤30ppm, about ≤27ppm, about ≤22.7ppm, about ≤20ppm or ≤10ppm.

[0139] In one implementation, at the calibration time, the apparent specific activity of the drug product is ≥10 mCi / mg, about ≥20 mCi / mg, about ≥30 mCi / mg, about ≥40 mCi / mg, about ≥50 mCi / mg, about ≥60 mCi / mg, about ≥70 mCi / mg, about ≥80 mCi / mg, or ≥90 mCi / mg DOTATATE and related substances.

[0140] In another embodiment, the average specific activity of the pharmaceutical product is about 2.96 MBq / μg. In another embodiment, the average specific activity of the pharmaceutical product is about 1.0 MBq / μg to about 5.0 MBq / μg. In another embodiment, the average specific activity of the pharmaceutical product is about 2.0 MBq / μg to about 4.0 MBq / μg. In another embodiment, the average specific activity of the pharmaceutical product is about 2.5 MBq / μg to about 3.5 MBq / μg. In another embodiment, at the calibration time, the average specific activity of the drug product 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.

[0141] In another embodiment, the filling 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 another embodiment, the filling 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.

[0142] In another embodiment, the pH of the pharmaceutical 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 pharmaceutical 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.

[0143] In one implementation scheme, 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%.

[0144] In another embodiment, ethanol is present in the pharmaceutical product in amounts 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 6%. In another embodiment, ethanol is present in the pharmaceutical product in amounts of about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, or about 10%.

[0145] In one embodiment, the ascorbic acid content in the pharmaceutical 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 pharmaceutical 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.

[0146] In one implementation, the RCP in the pharmaceutical product is ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%.

[0147] In another embodiment, the pharmaceutical product has a separated 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).

[0148] In one implementation, the pharmaceutical product is tested for filter integrity. In another implementation, the pharmaceutical product is tested for sterility.

[0149] iv. Drug product dosage

[0150] Regarding the dosage of the pharmaceutical products used in this article, this disclosure provides effective amounts. 64 Cu-DOTATATE is sufficient to enable positron emission tomography (PET) imaging in subjects who require it.

[0151] In one implementation, on the calibration date and time, the dose of the drug product administered to the subject in need is approximately 20 MBq to approximately 350 MBq, approximately 30 MBq to approximately 340 MBq, approximately 40 MBq to approximately 330 MBq, approximately 50 MBq to approximately 320 MBq, approximately 60 MBq to approximately 310 MBq, approximately 70 MBq to approximately 300 MBq, approximately 80 MBq to approximately 290 MBq, approximately 90 MBq to approximately 280 MBq, approximately 100 MBq to approximately 270 MBq, approximately 110 MBq to approximately 260 MBq, approximately 132 MBq to approximately 163 MBq, or approximately 111 MBq to 185 MBq, or approximately 120 MBq to 250 MBq.

[0152] In another implementation, on the calibration date and time, the dose of the drug product administered to the subjects in need is approximately 20 MBq, approximately 30 MBq, approximately 37 MBq, approximately 40 MBq, approximately 50 MBq, approximately 60 MBq, approximately 70 MBq, approximately 80 MBq, approximately 90 MBq, approximately 100 MBq, approximately 110 MBq, approximately 111 MBq, approximately 120 MBq, approximately 130 MBq, approximately 140 MBq, approximately 148 MBq, approximately 150 MBq, and approximately... 160MBq, approximately 170MBq, approximately 180MBq, approximately 185MBq, approximately 190MBq, approximately 200MBq, approximately 210MBq, approximately 220MBq, approximately 230MBq, approximately 240MBq, approximately 250MBq, approximately 260MBq, approximately 270MBq, approximately 280MBq, approximately 290MBq, approximately 300MBq, approximately 310MBq, approximately 320MBq, approximately 330MBq, approximately 340MBq, or 350MBq.

[0153] In one implementation, on the calibration date and time, the dosage of the drug product administered to the subjects in need is approximately 0.5 mCi to approximately 9.5 mCi, approximately 0.54 mCi to approximately 9.0 mCi, approximately 0.6 mCi to approximately 8.5 mCi, approximately 0.7 mCi to approximately 8.0 mCi, approximately 0.8 mCi to approximately 7.5 mCi, approximately 0.9 mCi to approximately 7 mCi, and approximately 1.0 mCi to approximately 6.5 mCi. i. 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.

[0154] In another implementation, on the calibration date and time, the dose of the drug product administered to the subjects in need is 0.1 mCi, approximately 0.5 mCi, approximately 0.54 mCi, approximately 0.6 mCi, approximately 0.7 mCi, approximately 0.8 mCi, approximately 0.9 mCi, approximately 1.0 mCi, approximately 1.1 mCi, approximately 1.2 mCi, approximately 1.3 mCi, approximately 1.4 mCi, approximately 1.5 mCi, approximately 2.0 mCi, approximately 2.5 mCi, approximately 3.0 mCi, approximately 3.1 mCi, approximately 3.2 mCi, approximately 3.3 mCi, approximately 3.4 mCi, and approximately 3.5 mCi. i, approximately 3.6 mCi, approximately 3.7 mCi, approximately 3.8 mCi, approximately 3.9 mCi, approximately 4.0 mCi, approximately 4.1 mCi, approximately 4.2 mCi, approximately 4.3 mCi, approximately 4.4 mCi, approximately 4.5 mCi, approximately 4.6 mCi, approximately 4.7 mCi, approximately 4.8 mCi, approximately 4.9 mCi, approximately 5.0 mCi, approximately 5.5 mCi, approximately 6.0 mCi, approximately 6.5 mCi, approximately 7.0 mCi, approximately 7.5 mCi, approximately 8.0 mCi, approximately 8.5 mCi, approximately 9.0 mCi, approximately 9.5 mCi, or approximately 10.0 mCi.

[0155] In one embodiment, the dosage of the pharmaceutical product is administered intravenously. In another embodiment, the pharmaceutical product is introduced intravenously into the subject of need in the form of a single dose, two doses, three doses, or multiple doses.

[0156] In one specific implementation, the drug product is administered to the subject via intravenous bolus at a dose of approximately 148 MBq (or approximately 4 mCi), and images are acquired approximately 45 to approximately 90 minutes after administration.

[0157] Dosage selection for elderly patients should be cautious, usually starting at the lower end of the dose range, reflecting decreased liver, kidney, or cardiac function, as well as a higher frequency of comorbidities or other drug treatments.

[0158] In one implementation, the drug product is administered to the subject over time intervals of approximately 15 minutes, approximately 10 minutes, approximately 5 minutes, approximately 4 minutes, approximately 3 minutes, approximately 2 minutes, or approximately 1 minute.

[0159] In one specific implementation, the amount of radioactivity used for adult PET imaging is 148 MBq (4 mCi) administered intravenously over a period of approximately 1 minute.

[0160] In one particular implementation, the pharmaceutical product contains 148 MBq (4 mCi) in a single-dose vial during calibration, at a concentration of 37 MBq (1 mCi) / 1 mL. 64 Cu-DOTATATE.

[0161] v. imaging

[0162] Somatostatin analogues and 64 Cu-DOTATATE binds competitively to the same somatostatin receptor and may affect imaging. Imaging should be performed before patients take a somatostatin analogue. For patients taking long-acting somatostatin analogues, a 28-day washout period before imaging is recommended. For patients taking short-acting somatostatin analogues, a 2-day washout period before imaging is recommended.

[0163] For PET imaging of pharmaceutical products, whole-body acquisition from the apex of the skull to the mid-thigh is recommended. Image acquisition should begin approximately 45 to 90 minutes after intravenous administration of the pharmaceutical product. The drug uptake time and scan duration should be adjusted based on the equipment used, as well as the characteristics of the patient and the tumor, to obtain optimal image quality.

[0164] G. Methods of administering pharmaceutical products

[0165] 64 Cu-DOTATATE binds to the somatostatin receptor. Based on signal intensity, it is used... 64 PET images obtained from Cu-DOTATATE injection show the presence and density of somatostatin receptors in the tissue. Uptake can also be seen in various non-NET tumors containing somatostatin receptors or as normal physiological variants. Tumors in NETs that do not carry somatostatin receptors will not be visualized.

[0166] The method of administering a drug product to a patient includes the following steps:

[0167] (a) Calibration 64 Cu-DOTATATE injection

[0168] (b) Use approximately 2 hours after calibration time. 64 Cu-DOTATATE injection

[0169] (c) During extraction and application 64 Aseptic techniques and radiation shielding are used when preparing Cu-DOTATATE injection.

[0170] (d) Visual inspection before application 64 The particulate matter and discoloration of Cu-DOTATATE injection solution should be considered; it can only be used if the solution is free of particulate matter or does not change color.

[0171] (e) Calculate the required application volume based on the measured activity, volume, calibration time, and date.

[0172] (f) Measure the patient's dose immediately before administering the drug product using a dose calibrator.

[0173] (g) Injection 64 Following Cu-DOTATATE injection, the patient was given an intravenous injection of 0.9% sodium chloride injection, USP, and...

[0174] (h) Any unused medications are disposed of safely in accordance with applicable regulations.

[0175] Intravenous injection 64 Table 4 shows the estimated radiation absorbed dose for each injection activity in organs and tissues of adult patients after Cu-DOTATATE injection.

[0176] Table 4. 64 Estimated absorbed radiation dose of Cu-DOTATATE injection per injection activity in selected organs

[0177]

[0178]

[0179] *The average value of 5 patients.

[0180] The effective radiation dose from administering 148 MBq (4 mCi) to an adult is approximately 4.7 mSv. For the active dose of 148 MBq (4 mCi), the typical radiation doses to key organs such as the 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, patients who have undergone splenectomy may experience higher uptakes and radiation doses to other organs or pathological tissues.

[0181] Non-radioactive somatostatin analogues and 64 Cu-DOTATATE competitively binds to the somatostatin receptor (SSTR2). Imaging is performed on patients before they take a somatostatin analogue. For patients taking long-acting somatostatin analogues, a 28-day washout period is recommended before imaging. For patients taking short-acting somatostatin analogues, a 2-day washout period is recommended before imaging.

[0182] 64 Cu-DOTATATE uptake reflects the level of somatostatin receptor density in NETs; however, uptake can also be seen in various other tumors that express the same somatostatin receptor. Increased uptake may also occur in other non-cancerous pathological conditions expressing the somatostatin receptor, including thyroid disease or subacute inflammation, or it may occur as a normal physiological variant (e.g., the uncinate process of the pancreas).

[0183] A negative scan performed on a patient without a history of NET after taking the medication does not rule out the disease.

[0184] Single dose administration 64 Maximum radioactivity was observed in the adrenal glands, kidneys, pituitary gland, spleen, and liver 1 to 3 hours after Cu-DOTATATE injection.

[0185] The single intravenous injection dose (4.15 ± 0.13 mCi) 64 Following the administration of Cu-DOTATATE injection (n=6), 16% to 40% of the injected dose of radioactivity was recovered from the urine within a 6-hour collection period.

[0186] In one implementation, at a single intravenous injection dose of 64 Following Cu-DOTATATE injection, approximately 5%, 10%, 15%, 16%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% of the injected dose of radioactivity are recovered in urine within a 6-hour collection period.

[0187] In another implementation, in a single intravenous injection dose of 64 Following the injection of Cu-DOTATATE, approximately 5%, 10%, 15%, 16%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% of the injected dose of radioactivity were recovered in urine within a 5-hour collection period.

[0188] In another implementation, in a single intravenous injection dose of 64 Following Cu-DOTATATE injection, approximately 5%, 10%, 15%, 16%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% of the injected dose of radioactivity are recovered in urine within a 4-hour collection period.

[0189] In one implementation, at a single intravenous injection dose of 64 Following the injection of Cu-DOTATATE, approximately 5%, 10%, 15%, 16%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% of the injected dose of radioactivity were recovered in urine within a 3-hour collection period.

[0190] In another implementation, in a single intravenous injection dose of 64 Following the injection of Cu-DOTATATE, approximately 5%, 10%, 15%, 16%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% of the injected dose of radioactivity were recovered in urine within a 2-hour collection period.

[0191] In another implementation, in a single intravenous injection dose of 64 Following the injection of Cu-DOTATATE, approximately 5%, 10%, 15%, 16%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% of the injected dose of radioactivity are recovered in urine within a 1-hour collection period. Example

[0192] The following examples provide methods for preparing high-purity 64 An improved process for Cu-labeled DOTATATE. By labeling DOTATATE with copper at low temperatures (i.e., ≤30°C), the chelation of DOTATATE with other metals can be reduced, thereby providing a pharmaceutical product with higher purity. Furthermore, these examples provide a method for expanding… 64 A useful method for producing Cu-DOTATATE while maintaining sufficient chemical stability to distribute the pharmaceutical product to patients. The following buffer solutions are prepared as used in Examples 1-5 described below.

[0193] Sodium acetate / gentianic acid buffer: Dissolve gentianic acid (GA) and sodium acetate (NaOAc) in high resistivity water (HRW), and adjust the pH of the resulting solution using glacial acetic acid or 1M sodium hydroxide. Further dilute the solution with HRW to obtain the desired concentrations of NaOAc and GA.

[0194] Sodium ascorbate buffer: Dissolve sodium ascorbate in HRW. Adjust the pH of the solution to 6.5-7.5 using 1M HCl or 1M NaOH. Further dilute the solution with HRW to obtain the desired concentration of sodium ascorbate.

[0195] Sodium ascorbate / ethanol buffer: Dissolve sodium ascorbate in HRW and anhydrous ethanol. Adjust the pH to 6.5-7.5 with 1M HCl or 1M NaOH, then further dilute with HRW to achieve the desired final concentration of sodium ascorbate and ethanol.

[0196] HPLC was performed using an Agilent 1200 series system equipped with a variable wavelength UV-Vis detector, subsequently connected in series with a sodium iodide detector (Bioscan B-FC-200P). 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% trifluoroacetic 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.

[0197] Example 1: Preparation of non-radioactive Cu-DOTATATE

[0198] Cu-DOTATATE was prepared by an initial non-radioactive reaction involving a solution of CuCl2 in 0.05 M HCl and a solution of DOTATATE peptide in a gentianic acid / sodium acetate buffer solution. The pH of the buffer solution was 6, unless otherwise specified. The formation of Cu-DOTATATE was confirmed by HPLC. The relative retention times of the DOTATATE starting material, the Cu-DOTATATE product peak, and other reactants were determined. Figure 2 The image shows a representative column chromatogram of a standard solution containing gentianic acid and DOTATATE.

[0199] In the initial experiments, approximately equimolar amounts of DOTATATE (0.035 μmol) in NaOAc / GA buffer and copper cations (0.039 μmol) in 0.05 M HCl were mixed in vials at room temperature. The reaction mixture was analyzed by HPLC at multiple time intervals. Figure 3 The HPLC column chromatogram of the sample collected at 5 minutes, shown in the figure, reveals a new product peak identified as Cu-DOTATATE at approximately 7.2 minutes, corresponding to a decrease in the DOTATE precursor peak (retention time approximately 6 minutes). The data indicate that the formation of Cu-DOTATATE is rapid and quantitative in a reaction mixture containing equimolar amounts of starting material at ambient temperature.

[0200] Example 2 – The formation of metal dotate complexes favors copper over other common metals

[0201] Due to the high specific activity of Cu-64, in typical [ 64 Cu]Cu 2+ The solution may contain nano- to microgram amounts of Cu.2+ [ 64 Cu]Cu 2+ Other trace metals that may be present in the solution are usually environmental impurities introduced during the manufacturing process. Common transition metals that may be present include iron, lead, zinc, and nickel. In these experiments, [the following methods were used]. nat A non-radioactive solution of Cu as 64 To evaluate the potential impact of metallic impurities on the preparation of copper Cu 64Dotatate, alternatives to Cu were selected because... nat Cu and 64 Cu is chemically identical.

[0202] At room temperature (~22℃), a 0.05M HCl solution containing 0.44 μg (0.00692 μmol) Cu was mixed with 100 μg (0.0693 μmol) DOTATATE in sodium acetate / gentianic acid buffer to achieve a DOTATATE to Cu molar ratio of 10:1. The reaction was monitored by HPLC. As shown in Table 5, at time points of 5 minutes and 7 hours, the HPLC peak areas of DOTATATE and Cu-DOTATE compounds showed essentially no change, indicating that the formation of Cu-DOTATATE is rapid and complete after 5 minutes at room temperature.

[0203] Table 5. Peak areas of DOTATATE and Cu-DOTATATE as a function of reaction time.

[0204]

[0205] Due to isotope enrichment 64 Ni is typically used in the production of 64 Cu, and therefore Ni, is another potential metallic impurity. However, due to the difficulty in achieving HPLC baseline resolution between Ni-DOATATE and Cu-DOATATE, similar competing experiments were conducted using Ni as a substitute for Cu to evaluate the reaction kinetics of Ni. The non-radioactive labeling reaction was carried out at room temperature (~22 °C) 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 gentian acid / sodium acetate buffer. No copper was added to the solution to better evaluate the chelating behavior of Ni in the presence of Fe, approximately Zn, and Co.

[0206] The reaction mixture was analyzed by HPLC at 5 minutes and 6 hours. Table 6 summarizes the peak area results and indicates that Ni 2+The reaction kinetics of Cu with DOTATATE 2+ Much slower. Therefore, similar to other transition metals (such as Fe, Co, Zn), at ambient temperature, the chelation of Cu by DOTATATE appears to occur faster than that of Ni.

[0207] Table 6. HPLC analysis results of Experiment 9 (peak areas of DOTATATE and metal-DOTATATE compounds)

[0208]

[0209] Example 3 – Formation of Cu-DOTATATE at a lower temperature

[0210] Experiments similar to those described in Example 2 were performed to determine whether Cu-DOTATATE formation occurred similarly at reduced temperatures (i.e., 15°C to 18°C). The reaction mixture was prepared as described in Example 2, but adjusted as needed to meet the conditions listed in Table 7.

[0211] Table 7. Marking conditions tested at reduced temperatures

[0212]

[0213] 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 DOTATATE labeling is essentially complete after 5 minutes, even at lower temperatures and in the presence of Fe.

[0214] Table 8. HPLC analysis of Cu-DOTATATE reaction carried out at reduced temperatures

[0215]

[0216] Example 4 – Preparation of up to 2000 mCi in a single reaction 64 Cu-DOTATATE

[0217] The radiolabeling reaction follows the general procedure described herein. 64 A solution of Cu in 0.05 M HCl was mixed with a solution of DOTATATE in sodium acetate / gentianic acid buffer. The reaction mixture was heated to 30 °C for 5 minutes, and then purified by C-18 solid-phase extraction. The purified... 64 Cu-DOTATATE was collected in 2 mL of 50% EtOH and then diluted with ascorbic acid solution. The activity of the final product was determined, and the radiochemical purity (RCP) was assessed by radio-HPLC analysis. Compared with non-radioactive DOTATATE (t...R =6.3 minutes) and Cu-DOTATATE(t R The standard sample (7.3 minutes) was used for characterization.

[0218] Typically, for a single reaction, scale-up reactions of Cu-64 were achieved from 100 mCi to approximately 7000 mCi. Representative reactions and results for batch sizes from 100 mCi to 7000 mCi are summarized in Table 9. HPLC analysis shows the retention times (t) of the major product peaks. R The reaction time was approximately 7.4 minutes, using non-radioactive Cu-DOTATATE (t R =7.3 minutes) co-eluted. Residual activity eluted (t) R =6.2–7.2 minutes) attributed to degradation products from radioactive decomposition. The chemical stability of the purified reaction solution was monitored by HPLC and demonstrated... 64 Cu-DOTATATE is stable for at least 47 hours, as shown in Table 9.

[0219] Table 9. Representative values ​​of approximately 100-2000 mCi 64 Overview of the Cu-DOTATATE reaction

[0220]

[0221] NP = Not performed

[0222] a Decay of TOS

[0223] b Sep-Pak purification material RCP

[0224] c The average of two samples

[0225] Example 5 – Preparation of over 7500 mCi 64 Cu-DOTATATE

[0226] Higher batch sizes can be produced by combining two sub-batch batches. 64 Cu-DOTATATE. For example, it is performed using 5250 mCi. 64 Cu(R1) and 4800mCi 64 Two radiolabeled reactions consisting of Cu(R2) were used to prepare copper Cu 64dotatate with a total concentration of approximately 9 Ci (without decay correction based on synthesis time). This was achieved by... 64 A solution of Cu in 0.05 M HCl and a solution of DOTATATE in gentian acid / sodium ascorbate buffer were used at a concentration of 1 mCi during synthesis. 64Cu 0.6 μg DOTATATE was mixed in a ratio of 0.6 μg and subjected to two separate 5Ci tests. 64 Cu radiolabeling reaction. The purified drug product solutions from each radiolabeling reaction were combined and diluted to provide a total of approximately 9 Ci at the time of purification. 64 Cu-DOTATATE. Process yield ≥95%. RCP of the final drug product solution at release ≥96%.

[0227] Example 6 – The largest prepared under conditions of reduced DOTATATE concentration 64 Cu-DOTATATE batches

[0228] exist 64 In the previous preparation of Cu-DOTATATE, the DOTATATE ligand was prepared at a concentration of 1 mCi 64 A ratio of 1 μg DOTATATE in Cu (i.e., a molar ratio of approximately 170:1 during synthesis) was added to the reaction mixture. This was done to improve the final... 64 The molar activity of Cu-DOTATATE prompted process improvements to reduce the amount of DOTATATE in the radiolabeling reaction. Two radiolabeling reactions (reaction 1 (R1) and reaction 2 (R2)) were carried out at up to 5250 mCi. 64 Cu and up to 4800mCi 64 The reaction was carried out under Cu. For R1, the total amount of labeled DOTATATE was approximately 3,125 μg or a concentration ≥276 μg / mL. For R2, the total amount of labeled DOTATATE was approximately 3,018 μg or a concentration ≥265 μg / mL. The ligand (i.e., the molar number of DOTATATE) and the radionuclide (i.e., the...) in the reaction mixture... 64 The molar ratios of Cu moles were approximately 102:1 (R1) and approximately 99:1 (R2). For each reaction, the mass (μg) of the ligand was related to... 64 The radioactivity ratio of Cu(mCi) concentration is approximately 0.6 μg / mCi. This will be measured in 0.05 M HCl. 64 The Cu solution was combined with the DOTATATE solution in sodium acetate / gentianic acid buffer. The radioactivity concentrations (RAC) of R1 and R2 at the time of radiosynthesis were ≥460 mCi / mL and ≥421 mCi / mL, respectively. The reaction was maintained at 30 °C for 5 min and at ambient temperature for 5 min prior to purification.

[0229] The crude reaction mixtures of R1 and R2 were purified separately using a C-18 solid-phase extraction (SPE) cartridge, and the eluates containing the purified products were combined to prepare a solution containing approximately 8.7 Ci. 64Bulk solution of Cu-DOTATATE (without decay correction based on synthesis time). Process yield ≥95%, and radiochemical purity (RCP) of the final drug product solution after dilution ≥96%.

[0230] In another experiment, approximately 7 Ci of 64Cu-DOTATATE was prepared in a single reaction using a Cu-64 DOTATATE ratio of 0.6 μg / mCi (see Table 9).

[0231] Example 7 – Gentianic acid and ethanol 64 Impact of Cu-DOTATATE Product Stability

[0232] The effects of ethanol (EtOH) and gentic acid (GA) content in the final dose matrix on the final dose were evaluated. 64 The effect of Cu-dotatate on chemical stability. In these experiments, a 500 mCi reaction was carried out using the general procedure outlined in Example 2, and 2 mL of 50% EtOH(aq) was used to... 64 The Cu-DOTATATE product was eluted from Sep-Pak into 5 mL of 50 mg / mL NaOAsc buffer and used as the purified product. 64 Cu-DOTATATE stock solution. From 64 In Cu-DOTATATE stock solution, transfer 1 mL of aliquots to vials containing the following solutions:

[0233] small bottle solution 1 2mL 66.3mg / mL NaOAsc 2 2mL 132.3mg / mL NaOAsc 3 2mL 66.3mg / mL NaOAsc + 8% EtOH 4 2mL 66.3mg / mL NaOAsc+15mg / mL GA 5 2mL 66.3mg / mL NaOAsc+30mg / mL GA 6 4mL 132.3mg / mL NaOAsc 7 5mL 64.8mg / mL NaOAsc+2.8%EtOH 8 2 mL of 49.5 mg / mL NaOAsc 9 2mL 94.5mg / mL NaOAsc 10 8mL 46.1mg / mL NaOAsc+3.5%EtOH 11 8mL 79.9mg / mL NaOAsc+3.5%EtOH 12 5mL 64.8mg / mL NaOAsc+2.8%EtOH

[0234] The stability of each vial was analyzed by HPLC, and the results are summarized in Table 10. Surprisingly, the highest degradation rates (24%–38%) were observed in those containing large amounts of GA, which is generally considered a radioprotectant. In these samples, at t R =At 6.6 minutes and 7.1 minutes, 64 Reduction and Free Cu-DOTATATE 64 The increase in Cu was associated with two unknown radioactive impurities. Since vials with similar activity concentrations showed almost no degradation, radioactive decomposition is unlikely to have led to degradation; therefore, the mechanism leading to chemical instability remains unknown. [The instability occurred within 48 hours.] 64 The only other condition where Cu-DOTATATE loss exceeded 2% was vial 3, which differed from the control (via vial 1) in that it contained approximately 10% EtOH.

[0235] Table 10. Under various conditions, over time, the complete... 64 Percentage of Cu-DOTATATE.

[0236]

[0237] a Test 1-2 hours after TOS; b Test 24-26 hours after TOS; c Tests were conducted 45-48 hours after TOS.

[0238] The results showed that in solutions containing sodium ascorbate at concentrations of 45-122 mg / mL, ethanol at concentrations of 1.6%-5.2%, and an active concentration of 3.6-16 mCi / mL (during preparation), 64 Cu-DOTATATE remained chemically stable for 2 days at a purity greater than 95%. For solutions containing sodium ascorbate at concentrations up to 98 mg / mL, ethanol at concentrations up to 9.7%, and an activity concentration of approximately 18 mCi / mL (at preparation time), 64 The Cu-dotatate product remains stable at a purity greater than 90%.

[0239] Example 8: Preparation of Cu-DOTATATE in the presence of increased gentianic acid or sodium ascorbate

[0240] The general reaction protocol used in previous experiments was repeated, except that the concentration of gentianic acid in the reaction mixture was increased fourfold. After the reaction, the mixture was sampled and purified using a C-18 solid-phase extraction (SPE) cartridge, and the purified product was analyzed by HPLC to determine the reaction yield. The results of the HPLC analysis are summarized in Table 11. Quantitative recovery of DOTATATE and Cu-DOTATATE was almost achieved, and the labeling efficiency and purification were unaffected by the large excess of gentianic acid in the reaction mixture.

[0241] Table 11. Summary of HPLC analysis of Cu-DOTATATE formulations

[0242]

[0243] Gentian acid in the reaction mixture acts as a radioprotectant and helps reduce radioactive degradation. To assess the possibility of using another radioprotectant, a reaction was carried out in which sodium ascorbate was added to the reaction mixture (pH = 6.8). Dotate (0.0693 μmol) in the sodium acetate / gentian acid buffer was reacted with Cu... 2+ The solutions were mixed in 0.05 M HCl (0.0069 μmol) and diluted with sodium ascorbate to allow the DOTATATE to react with Cu. 2+The ratio was 10:1. The reaction mixture was mixed at room temperature, and samples were taken at 5 minutes and 51 minutes to monitor the formation of Cu-DOTATATE by HPLC analysis. The HPLC peak area of ​​Cu-DOTATATE was 2.33 mV / min at 5 minutes and 1.36 mV / min at 51 minutes, indicating that the reaction was completed before 5 minutes.

[0244] Example 9—Non-radioactive copper pair 64 Effect of Cu-DOTATATE on Radiochemical Purity

[0245] Three reactions were performed, each using approximately 5 Ci. R1 was a control reaction without the addition of non-radioactive copper. Copper was not detected in R1. Non-radioactive copper was added to R2 and R3 to investigate its effect on RCP. The total copper content in R2 was approximately 139 μg in 12.6 mL (11.0 μg / mL). The total copper content in R3 was approximately 476 μg in 15.3 mL (31.1 μg / mL). The total DOTATATE labeled in each reaction was approximately 3000 μg (R1), approximately 3000 μg (R2), and approximately 3600 μg (R3), or concentrations ≥250 μg / mL, approximately ≥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 then purified and diluted to its final bulk solution.

[0246] The reactivity ratios (RACs) of the final drug product solutions R1, R2, and R3 were 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 for R1, R2, and R3 were approximately 95.4%, approximately 98.7%, and approximately 95.6%, respectively. The relative productivity (RCP) of each diluted final drug product solution was ≥95.5% (R1), approximately ≥97.3% (R2), and ≥97.9% (R3).

[0247] Example 10 – Recovery of DOTATATE from SPE cartridges at higher flow rates

[0248] Typically, the flow rate through the SPE cartridge is kept low (i.e., 1-5 mL / min) to ensure sufficient desired product loading on the cartridge and to ensure a high reflux rate of the purified product eluent. 64In the case of Cu-DOTATATE, concentrating the product on an SPE column can lead to higher radioactive damage, especially for high-activity batches. Therefore, the regeneration rate of DOTATATE was evaluated to reduce purification time and achieve higher flow rates. Since C-18 SPE chemistry is primarily driven by the interaction between DOTATATE and the cartridge, experiments were conducted using a non-radioactive solution of DOTATATE, as Cu-DOTATATE or other metal DOTATATE species behave very similarly.

[0249] A solution of DOTATATE in sodium acetate / gentianic acid buffer was prepared and loaded onto a C-18 SPE column at a flow rate of 12 mL / min or 18 mL / min. The column was rinsed with water and DOTATATE was eluted in 50% EtOH. The amount of DOTATATE in the loading solution and the eluent of the purified product was evaluated by HPLC. At a flow rate of 12 mL / min, 5.3% of DOTATATE permeated the SPE column during loading, while 97.1% of DOTATATE was recovered in the eluent (total recovery rate 102%). When purification was performed at 18 mL / min, 28.4% of DOTATATE permeated during loading, and 68.7% was recovered in the eluent (total recovery rate 97%). The results indicate that a flow rate of at least 12 mL / min can be used for purification. 64 Cu-DOTATATE was produced while maintaining a near-quantitative recovery rate. The loading solution was collected in fractions, and the DOTATATE recovery results for each fraction are shown below. Figure 4 and Figure 5 As shown.

[0250] Example 11 – Improving the purification yield of Cu-DOTATATE using 50% ethanol eluent

[0251] Radiolabeled copper Cu-64 Dotatate is typically purified using a 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 usually eluted from the cartridge using 100% ethanol. 64 Cu-DOTATATE compounds. We have found that using 50% EtOH can improve the purification yield of Cu-DOTATATE. Several experiments were conducted to support this observation.

[0252] For each condition, prepare three portions of the reaction mixture containing copper (2+) ions, transition metal ion impurities, and a bioconjugated chelating agent (DOTATATE). Incubate the reaction mixture at room temperature (approximately 20°C) for 5 minutes, then purify using a C-18 SPE column and elute with either 100% EtOH (n=3) or 50% EtOH (n=3). The ratio of DOTATATE to metal in the reaction mixture is provided in Table 12.

[0253] Table 12. Molar ratio of DOTA-TATE to metal ions used for labeling

[0254] metal ions DOTA-TATE: Metal Molar Ratio Cu(2+) 4:1 Fe(3+) 4:1 Co(3+) 8:1 Pb(2+) 8:1 Total metal content 1.3:1

[0255] Approximately 10 minutes after mixing, a sample of the crude reaction mixture was analyzed by HPLC to obtain the in-situ reaction yield. Each reaction mixture was then purified using a C-18 SPE column, with the product eluted with 100% EtOH or 50% EtOH. The separation yield of the purified product solution was determined by HPLC analysis. The results are shown in Table 13. The reaction yield was determined by comparison with DOTATATE standards.

[0256] Table 13. Comparison of Cu-DOTATATE separation yields using 50% EtOH or 100% EtOH eluent

[0257]

[0258] 1 The average value of preparations with n=3. 2 The Fe / Co-DOTATATE peak was not baseline resolved and integrated into a single peak.

[0259] Example 12 – The Efficacy of Drug Products

[0260] The efficacy of the drug product was determined in two single-center, open-label studies. Study 1 prospectively evaluated a total of 63 participants, including 42 patients with known or suspected NET based on routine histological imaging or clinical assessment and 21 healthy volunteers. Of the 42 patients, 37 (88%) had a history of NET at the time of imaging with the drug product. In the total study population of 63 participants, 28 (44%) were male and 35 (56%) were female, with the majority being Caucasian (86%). The mean age of the participants was 54 years (range 25 to 82 years).

[0261] Drug product images from each subject were interpreted as NET positive or negative by three independent interpreters who were blinded to clinical information and other imaging results. PET imaging results were compared to a comprehensive reference standard, which was blinded by a single oncologist's assessment of the subject's diagnosis based on existing histopathological findings, and reported on routine imaging (MRI, contrast CT, bone scintigraphy, etc.) performed within 8 weeks prior to drug imaging. 18 F] Fluorodeoxyglucose PET / CT, [ 18 F] Sodium fluoride PET / CT, [ 111 In] Pentapeptide Indium SPECT / CT, [ 68 Ga-dotatate PET / CT was used, along with reports including clinical and laboratory data such as chromogranin A and serotonin levels. The proportion of disease-positive subjects in each composite reference identified as positive by imaging with the drug product was used to quantify the percentage of positive concordance. In each composite reference, the proportion of disease-free subjects identified as negative by imaging with the drug product was used to quantify the percentage of negative concordance. Table 14 shows the performance of the drug product used in Study 1 in detecting NET.

[0262] Table 14. Performance of drug products in the detection NET by readers in Study 1

[0263]

[0264] n: Number of patients, CI: Confidence interval, *Reader 1 interprets one of the 63 PET scans as "not evaluable", **Wilson score interval with continuity correction.

[0265] Study 2 showed similar results through a retrospective analysis of published data from 112 patients with a known history of NET (63 men, 49 women; mean age 62 years, ranging from 30 to 84 years).

[0266] Example 13—Safety and Efficacy of Pharmaceutical Products

[0267] In the safety and efficacy trial, 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 in <2% of participants: (a) gastrointestinal disorders: nausea and vomiting; and (b) vascular disorders: flushing.

[0268] 126 patients with a known history of NET received a single dose 64 Cu-DOTATATE injection. It has been reported that four patients experienced nausea immediately after the injection.

[0269] The embodiments described herein are intended to be exemplary only. Those skilled in the art will understand that variations and modifications can be made without departing from the scope of the invention as covered by the following claims.

Claims

1. A method for positron emission tomography (PET) imaging comprising... 64 Cu-DOTATATE 64 Cu-N-[(4,7,10-tricarboxymethyl-1,4,7,10-tetraazacyclododecyl-1-yl)acetyl]-(D)-phenylalanyl-(L)-cysteyl-(L)-tyrosinyl-(D)-tryptophanyl-(L)-lysine-(L)-threonyl-(L)-cysteyl-(L)-threonyl-cyclic disulfide (Cys 2 -Cys 7 ) pharmaceutical products, among which, The drug product contains a total DOTATATE ratio of 125:1 to 1:1 and a total DOTATATE ratio of 125:1 to 1:

1. 64 The molar ratio of Cu, and on the calibration date and time, is 4 mL of solution containing 148 MBq (4 mCi). 64 Single-dose vials of Cu-DOTATATE, wherein the radiochemical purity of the drug product is ≥96%.

2. The pharmaceutical product according to claim 1, wherein, The radiochemical purity of the drug product is ≥97%.

3. The pharmaceutical product according to claim 1, wherein, The radiochemical purity of the drug product is ≥98%.

4. The pharmaceutical product according to claim 1, wherein, The radiochemical purity of the drug product is ≥99%.

5. The pharmaceutical product according to claim 1, wherein, The drug product is stable for 48 hours after preparation.

6. The pharmaceutical product according to claim 1, wherein, The drug product is stable for 24 hours after preparation.

7. The pharmaceutical product according to claim 1, wherein, Total DOTATATE and Total 64 The molar ratio of Cu is 125:1 to 75:1.

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