US20220387637A1

US20220387637A1 - Radiopharmaceutical and methods

Info

Publication number: US20220387637A1
Application number: US17/817,542
Authority: US
Inventors: Joe McCann; Justyna Kelly; Paul Billone
Original assignee: Centre for Probe Development and Commercialization
Current assignee: Centre for Probe Development and Commercialization
Priority date: 2020-08-27
Filing date: 2022-08-04
Publication date: 2022-12-08
Also published as: TW202227143A; AU2021330382A1; EP4204011A2; US11439714B2; US20220062447A1; JP2023540240A; WO2022043754A2; CN116528904A; AU2021330382A9; WO2022043754A3; CA3190855A1

Abstract

A radiopharmaceutical ¹⁷⁷Lu-DOTATATE compound, a composition, and a kit are provided. Further provided are methods of synthesis of a ¹⁷⁷Lu-DOTATATE compound and methods of treatment that comprise a ¹⁷⁷Lu-DOTATATE compound.

Description

The present application claims priority of U.S. provisional application No. 63/071,138 filed Aug. 27, 2020, which is incorporated by reference herein in its entirety.

FIELD

The radiopharmaceutical ¹⁷⁷Lu-DOTATATE (¹⁷⁷Lu-octreotate) and a pharmaceutical composition thereof are provided. Further provided are methods of preparing ¹⁷⁷Lu-DOTATATE compound and pharmaceutical compositions that include ¹⁷⁷Lu-DOTATATE.

BACKGROUND

Radiopharmaceuticals have been used for a variety of therapeutic and diagnostic indications. Among others, radiolabeled molecules have been useful to treat various malignant tumors.
Use of these agents presents certain challenges, including with respect to stability and shelf-life. In particular, therapeutic compositions comprising a radionuclide may undergo radiolysis during preparation and storage. During radiolysis, radionuclide emissions may react with other groups of the pharmaceutical agent thereby resulting in decomposition of the agent as well as undesired effects.
It thus would be desirable to have additional and improved radiopharmaceutical agents.

SUMMARY

We now provide, inter alia, radiopharmaceutical ¹⁷⁷Lu-DOTATATE compound, methods of preparing the compound, and pharmaceutical compositions and methods of treatment including ¹⁷⁷Lu-DOTATATE.
We have now found, inter alia, 1) new methods to produce high purity ¹⁷⁷Lu-DOTATATE including with high levels of ¹⁷⁷Lu incorporation, mild reaction temperatures and/or reduced reaction times and 2) new ¹⁷⁷Lu-DOTATATE pharmaceutical compositions that maintain radiochemical purity for extended storage times following preparation (e.g. >90 or 95%, 3, 4 or 5 days or more, 25° C.). See, for instance, the results of the Examples which follow.
The preferred methods and pharmaceutical compositions include relatively high amounts of one or more stabilizers, particularly high amounts of one or more ascorbate compounds and/or one or more gentisate compounds.
More particularly, pharmaceutical compositions are provided that comprise: (a) a complex of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE); and (b) one or more stabilizer compounds in a concentration of at least 25 mg/mL.
In certain aspects, suitably, the complex of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) is present in an amount of at least 5, 10, 15, 20, 25 or 30 mCi/mL.
In certain aspects, the one or more stabilizers comprise one or more ascorbate compounds.
In certain aspects, the one or more stabilizers comprise one or more gentisate compounds.
In certain preferred aspects, the pharmaceutical composition comprises two or more distinct stabilizers. In particular preferred aspects, the pharmaceutical composition comprises stabilizers that include at least one ascorbate compound and at least one gentisate compound.
In certain preferred aspects, one or more stabilizer compounds are present in a pharmaceutical composition in an amount of 30 mg/mL or greater.
In certain preferred aspects, one or more stabilizer compounds are present in a pharmaceutical composition in an amount of 40 mg/mL or greater.
In certain preferred aspects, one or more stabilizer compounds are present in a pharmaceutical composition in an amount of 50 mg/mL or greater.
In certain preferred aspects, one or more stabilizer compounds are present in a pharmaceutical composition in an amount of from about 25, 30, 35 or 40 mg/mL to 45, 50, 55, 60, 65, 70, 75 or 80 mg/mL.
In certain preferred aspects, a pharmaceutical composition comprises two distinct stabilizers, particularly 1) an ascorbate compounds such as an ascorbate salt and/or ascorbic acid and 2) a gentisate compound such as gentisic acid.
Suitably, the pharmaceutical composition is an aqueous formulation. In certain preferred aspects, a pharmaceutical composition does not contain an alcohol such as ethanol or other organic solvent. In certain preferred aspects, a pharmaceutical composition is at least substantially free (i.e. less than 5, 4, 3, 2, 1 or 0.5 weight percent based on total composition weight) of an alcohol such as ethanol or other organic solvent.
The compound tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) has the following structure 1:
¹⁷⁷Lu-DOTATATE is a lutetium-177 complex of the above compound 1 and may be represented by the following structure 2:
In an aspect, methods are provided for preparing ¹⁷⁷Lu-DOTATATE (structure 2 above), which includes admixing lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE, structure 1 above) in the presence of one or more stabilizer compounds and forming a complex of ¹⁷⁷Lu and DOTATATE. This reaction to form a complex of ¹⁷⁷Lu and DOTATATE (e.g. such complex including structure 2 above) that may include admixing ¹⁷⁷Lu and DOTATATE is sometimes referred to herein as an “incorporation reaction”.
The one or more stabilizer compounds are suitably present in an incorporation reaction and/or a pharmaceutical composition in an amount of 7 mg/mL or greater, including 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, 30, 35, 40, 45 or 50 mg/mL or greater.
In preferred aspects, the tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) is admixed with lutetium-177 as an aqueous formulation.
In further preferred aspects, the one or more stabilizer compounds are present in an incorporation reaction and/or a pharmaceutical composition in an amount of 12, 15, 20 or 25 mg/mL or greater. In certain aspects, the one or more stabilizer compounds are present in an incorporation reaction and/or a pharmaceutical composition in an in an amount of 30 mg/mL or greater. In additional aspects, the one or more stabilizers are present in an incorporation reaction and/or a pharmaceutical composition in an in an amount of 40 mg/mL or greater. In still further aspects, the one or more stabilizers are present in an incorporation reaction and/or a pharmaceutical composition in an in an amount of 50 mg/mL or greater.
In certain preferred aspects, each of two or more distinct stabilizer compounds are present in an incorporation reaction and/or a pharmaceutical composition in an amount of 10 mg/mL or greater.
In certain preferred aspects, the one or more stabilizers present in an incorporation reaction and/or a pharmaceutical composition include one or more ascorbate compounds. In certain preferred aspects, the one or more stabilizers present in an incorporation reaction and/or a pharmaceutical composition comprise one or more gentisate compounds.
In certain preferred aspects, the one or more stabilizers present in an incorporation reaction and/or a pharmaceutical composition include one or more gentisate compounds in an amount of at least 7, 8, 9 or 10 mg/mL and one or more ascorbate compounds in an amount of at least 15, 20, 25 or 30 mg/mL, or one or more gentisate compounds in an amount of at least 15 mg/mL and one or more ascorbate compounds in an amount of at least 40 mg/mL.
In certain aspects, at least one of the stabilizers present in an incorporation reaction and/or a pharmaceutical composition is an ascorbate compounds, particularly an ascorbate salt such as sodium ascorbate.
Preferably, prior to the incorporation reaction or addition to ¹⁷⁷Lu-DOTATATE to provide a pharmaceutical composition, the ascorbate compound such as sodium ascorbate is assessed for purity or absence of a material that may inhibit the incorporation reaction of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE). For instance, a sample from a lot of an ascorbate compound such as an ascorbate metal salt e.g. sodium ascorbate can be tested such as in a test-scale (small-scale) incorporation reaction of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) to ensure the ascorbate compound lot does not adversely impact the formation of the complex of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) or the radiochemical stability of the formed complex over time.
Accordingly, methods are provided that comprise determining purity of an ascorbate compound prior to admixing the ascorbate compound with lutetium-177 and/or tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) and/or a complex of lutetium-177 and/or tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE). Suitably, the ascorbate compound is determined to be effective in an incorporation reaction and method comprises admixing the effective ascorbate compound with lutetium-177 and/or tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) and/or a complex of lutetium-177 and/or tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE). In such methods, the ascorbate compound may be an ascorbate salt, such as sodium ascorbate.
In certain preferred aspects, the admixture of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) is heated. In particular aspects, the admixture is heated for 60 minutes or less, for 50 minutes or less, for 40 minutes or less, for 30 minutes or less, for 20 minutes or less, or for 15 minutes or less.
In certain preferred aspects, the admixture of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) is heated at about 80° C.±10° C., or at about 80° C.±5° C.
In certain preferred aspects, the present methods include cooling the admixture following heating.
In certain preferred aspects, following the admixing, the complex of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) is mixed with a formulation composition. Preferably, the formulation composition is an aqueous composition that includes one or more stabilizer compounds. The formulation composition suitably includes one or more stabilizer compounds present in an amount of 5, 6, 7, 8, 9 or 10 mg/mL or greater, in an amount of 20 mg/mL or greater, in an amount of 30 mg/mL or greater, in an amount of 40 mg/mL or greater, or in an amount of 50, 60, 70, 80 or 90 mg/mL or greater.
In certain preferred aspects, a formulation composition further comprises one or more sequestering agents.
In certain preferred aspects, incorporation of lutetium-177 into or with tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) to provide ¹⁷⁷Lu-DOTATATE is preferably greater than 98 mole percent, or greater than 99 mole percent based on the molar equivalent of lutetium-177 used in the incorporation reaction.
In certain preferred aspects, an acidic aqueous formulation of lutetium-177 is admixed with the tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE). Preferably, a hydrogen halide or acid halide aqueous formulation of lutetium-177 is admixed with the tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE). For example, a hydrochloride acid aqueous formulation of lutetium-177 is admixed with the tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE).
In an aspect, provided is ¹⁷⁷Lu-DOTATATE obtainable, or obtained by the methods described herein.
In an aspect, provided is a pharmaceutical composition including ¹⁷⁷Lu-DOTATATE as described herein. Preferably, radiochemical purity of the composition is 95% or greater for 3 days or more at 25° C., or 95% or greater for 4 days or more at 25° C., or 95% or greater for 5 days or more at 25° C. As referred to herein, radiochemical purity is preferably assessed via HPLC analysis.
In an aspect, provided is a single dosage kit including ¹⁷⁷Lu-DOTATATE or pharmaceutical composition described herein. Moreover, provided is a multiple dosage kit including the ¹⁷⁷Lu-DOTATATE or pharmaceutical composition described herein.
In an aspect, provided is a method of treating a subject suffering from cancer. The method includes administering to the subject an effective amount of ¹⁷⁷Lu-DOTATATE or pharmaceutical composition described herein.
In certain aspects, the subject is suffering from a neuroendocrine tumor. In certain aspects, the subject is suffering from neuroendocrine tumors that originate from foregut, hindgut, midgut, lung, ovary, medulla, adrenal medulla, adrenal, kidney, pituitary, thyroid or paraganglia.
In certain aspects, the subject is suffering from a gastroenteropancreatic neuroendocrine tumor (GEP-NET), foregut, midgut and hindgut neuroendocrine tumors.
In certain aspects, the subject is suffering from unresectable or metastatic neuroendocrine tumor(s).
In certain aspects, the methods include identifying a patient for treatment by assessing neuroendocrine tumors of the patient to be somatostatin receptor positive, for example with ⁶⁸Ga-DOTATATE and a positron emission tomography scan.
Use of the ¹⁷⁷Lu-DOTATATE compound and compositions to treat a disease or disorder as disclosed herein is also provided.
Further provided are methods for manufacture of a medicament that comprises the ¹⁷⁷Lu-DOTATATE compound and composition to treat a disease or disorder as disclosed herein.
Other aspects of the invention are disclosed infra.

BRIEF DESCRIPTION OF THE FIGURE

FIG. 1 shows exemplary processes for manufacturing ¹⁷⁷Lu-DOTATATE.

FIG. 2 shows a flow chart for an exemplary fabrication process of Example 1.

FIG. 3 shows effect of reaction time, temperature, and pH on % incorporation of Lu-177 as disclosed in Example 3.

FIG. 4 shows effect of sodium ascorbate and gentisic acid on % incorporation of Lu-177 as disclosed in Example 3.

FIG. 5 . Radio-HPLC chromatogram of sample 2 analysis (Example 3). Overlay of the UV (220 nm) with the radiation detector trace. ¹⁷⁷Lu-DOTATATE eluted at t_R=10.2 minutes and the precursor peptide eluted at t_R=11.4 minutes as disclosed in Example 3.

FIG. 6 (includes FIGS. 6A-6B). FIG. 6A: HPLC analysis of chemical impurities in reactions with various amounts of gentisic acid. FIG. 6B. HPLC analysis of chemical impurities in reaction heated at different temperatures. ¹⁷⁷Lu-DOTATATE eluted at t_R=10.2 minutes and the precursor peptide eluted at t_R=11.4 minutes.

FIG. 7 shows radio-HPLC spectra of day 5 formulation screening samples as disclosed in Example 3 which follows.

DETAILED DESCRIPTION

As discussed above, ¹⁷⁷Lu-DOTATATE is a lutetium-177 (¹⁷⁷Lu³⁺) complex of the compound 1
and may be represented by the following structure 2:
¹⁷⁷Lu-DOTATATE is also represented with a chemical (IUPAC) name of ¹⁷⁷Lu-tetraazacyclododecane tetra-acetic acid-octreotate or ¹⁷⁷Lu-4,7,10-Tricarboxymethyl-1,4,7,10-tetraaza-cyclododecan-1-yl-acetyl-D-Phe-Cys-Tyr-D-Trp-Lys-Thr-Cys-Thr-OH. ¹⁷⁷Lu-DOTATATE has a molecular formula of C₆₅H₉₀[¹⁷⁷Lu]N₁₄O₁₉S₂and a molecular weight (average molecular weight) of 1610.61 g/mol.
The present invention, including compounds, methods, and pharmaceutical compositions/formulations will be described with reference to the following definitions which, for convenience, are set forth below. Unless otherwise specified, the below terms used herein are defined as follows:

Definitions

As used herein, the term “a,” “an,” “the” and similar terms used in the context of the present invention (especially in the context of the claims) are to be construed to cover both the singular and plural unless otherwise indicated herein or clearly contradicted by the context.
The language “and/or” is used herein as a shorthand notation to represent the expression “and,” describing the combination of items, as well as “or,” describing the items in the alternative form.
Unless otherwise stated, structures depicted herein are also meant to include all stereochemical forms of the structure; i.e., the R and S configurations for each asymmetric center. Therefore, single stereochemical isomers as well as enantiomeric and diastereomeric mixtures of the present compounds are within the scope of the invention.
The term “about”, as used herein, means an acceptable margin of error for a particular value, which depends in part on how the value is measured or determined. In certain aspects, “about” as used herein will be understood by persons of ordinary skill in the art to mean up to plus or minus 20% of the particular term. In further aspects, “about” as used herein will be understood by persons of ordinary skill in the art to mean up to plus or minus 10% of the particular term.
As used herein, the term “substantially pure” means sufficiently homogeneous to appear free of readily detectable impurities as determined by standard analytical methods, such as thin layer chromatography (TLC), gel electrophoresis, high performance liquid chromatography (HPLC), nuclear magnetic resonance (NMR), and mass spectrometry (MS); or sufficiently pure such that further purification would not detectably alter the physical and chemical properties, or biological and pharmacological properties, such as enzymatic and biological activities, of the substance. In certain aspects, “substantially pure” refers to a collection of molecules, wherein at least about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 97%, about 98%, about 98.5%, about 99%, about 99.5% or about 99.9% or greater of the molecules are a single compound, including a racemic mixture or a single stereoisomer thereof, as determined by standard analytical methods.
As used herein, and unless otherwise specified, the terms “treat,” “treating” and “treatment” refer to the eradication or amelioration of a disease, disorder, or condition, or of one or more symptoms associated with the disease, disorder or condition. In certain aspects, the terms refer to minimizing the advancement or worsening of the disease, disorder, or condition resulting from the administration of a formulation of the invention to a patient with such a disease, disorder, or condition. In some aspects, the terms refer to the administration of a formulation provided herein, after the onset of symptoms of the particular disease, disorder, or condition. The terms “treat,” “treating”, “treatment”, or the like, as used herein covers the treatment of a disease, disorder, or condition in a subject, e.g., a mammal, and includes at least one of: (i) inhibiting the disease, disorder, or condition, i.e., partially or completely halting its progression; (ii) relieving the disease, disorder, or condition, i.e. causing regression of symptoms of the disease, disorder, or condition, or ameliorating a symptom of the disease, disorder, or condition; and (iii) reversal or regression of the disease, disorder, or condition, preferably eliminating or curing of the disease, disorder, or condition. In a particular embodiment the terms “treat,” “treating”, “treatment”, or the like, covers the treatment of a disease, disorder, or condition in a mammal, e.g., a primate, e.g., a human, and includes at least one of (i), (ii), and (iii) above. As is known in the art, adjustments for age, body weight, general health, sex, diet, time of administration, drug interaction and the severity of the condition may be necessary, and will be ascertainable with routine experimentation by one of ordinary skill in the art based on the invention described herein.
As used herein, the terms “subject”, and “patient” are used interchangeably. The terms “subject” and “patient” refer to an animal such as a mammal including non-primates (e.g., a cow, pig, horse, sheep, rabbit, guinea pig, rat, cat, dog, and mouse) and primates (e.g., a monkey, chimpanzee and a human). In a particular embodiment, the subject is a human.

Syntheses

In an aspect, ¹⁷⁷Lu-DOTATATE (including structure 2) can be prepared by complexing or incorporating ¹⁷⁷Lu (Lutetium 177) or halide thereof such as ¹⁷⁷LuCl₃with DOTATATE (including structure 1 below):
The compound 1 DOTATATE may be suitably formed as described previously such as in Raheem et al. Bioconjugate Chem. 2020. https://doi.org/10.1021/acs.bioconjchem.0c00325. The compound 1 DOTATATE is also commercially available.
It is understood that ¹⁷⁷Lu-DOTATATE as referred to herein includes the above structure 2 as well as other complexes of ¹⁷⁷Lu-DOTATATE. For instance, references herein to ¹⁷⁷Lu-DOTATATE include compounds that generally correspond to structure 2 but where the ¹⁷⁷Lu substantially complexes to other portions or moieties (such as one or more other nitrogens) of the DOTATATE molecule than as depicted in 2 above. References to ¹⁷⁷Lu-DOTATATE also may include other stereoisomers than those shown in 1 and 2 above, although the stereoisomer depicted in 1 and 2 is preferred.
To synthesize ¹⁷⁷Lu-DOTATATE, a salt form of lutetium-177 (¹⁷⁷Lu) can be admixed with DOTATATE in an incorporation reaction. The ¹⁷⁷Lu suitably may be carrier-added or more preferably no-carrier-added (n.c.a.) lutetium-177. To facilitate incorporation (e.g. complexing including chelating) of lutetium-177 with DOTATATE compound, preferably an admixture of the compounds is in the presence of one or more stabilizer compounds.
In an aspect, the method of preparing ¹⁷⁷Lu-DOTATATE includes a step of admixing lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) in the presence of one or more stabilizer compounds; and forming a complex of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE).
Preferably, as discussed, in an incorporation reaction, the one or more stabilizer compounds are present in an amount of 7, 8, 9 or 10 mg/mL or greater. In certain aspects, the one or more stabilizer compounds are present in an incorporation reaction in an amount of 20 mg/mL or greater. In certain aspects, the one or more stabilizer compounds are present in an incorporation reaction in an amount of 30 mg/mL or greater. In certain aspects, the one or more stabilizer compounds are present in an incorporation reaction in an amount of 40 mg/mL or greater. In certain aspects, the one or more stabilizer compounds are present in an incorporation reaction in an amount of 50, 60 or 70 mg/mL or greater.
Preferably, the one or more stabilizers comprise one or more ascorbate compounds. For example, the ascorbate compound may include an ascorbate salt such sodium ascorbate.
Preferably, the one or more stabilizers comprise one or more gentisate compounds. For example, the gentisate compound may include, but not limited to, gentisic acid.
Preferably, the one or more stabilizers in an incorporation reaction comprise one or more gentisate compounds in an amount of at least 5, 6, 7, 8, 9 or 10 mg/mL and one or more ascorbate compounds in an amount of at least 10, 20, 30, 40 or 50 mg/mL. In certain aspects, the one or more stabilizers in an incorporation reaction comprise one or more gentisate compounds in an amount of at least 15 mg/mL and one or more ascorbate compounds in an amount of at least 40 mg/mL.
Preferably, the admixture of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) is heated. In certain aspects, the admixture is heated for 20 minutes or less. In certain aspects, the admixture is heated for 15 minutes or less.
Preferably, the admixture of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) is heated at a temperature of about 80° C.±10° C. In certain aspects, the admixture lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) is heated at a temperature of about 80° C.±5° C.
The method further includes cooling the admixture of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) following heating.
In certain aspects, following the admixing, the complex of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) is mixed with a formulation composition. Preferably, the formulation composition is an aqueous composition that comprises one or more stabilizer compounds. In certain aspects, the formulation composition comprises one or more stabilizer compounds in an amount of 5, 6, 7, 8, 9 or 10 mg/mL or greater. In certain aspects, the formulation composition comprises one or more stabilizer compounds in an amount of 20 mg/mL or greater. In certain aspects, the formulation composition comprises one or more stabilizer compounds present in an amount of 30 mg/mL or greater. In certain aspects, the formulation composition comprises one or more stabilizer compounds present in an amount of 40 mg/mL or greater. In certain aspects, the formulation composition comprises one or more stabilizer compounds present in an amount of 50, 60 or 70 mg/mL or greater.
In certain aspects, the formulation composition further includes one or more sequestering agents.
Preferably, incorporation of lutetium-177 into or with tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) to provide ¹⁷⁷Lu-DOTATATE is greater than 98 mole percent based on the molar equivalent of lutetium-177 used in the incorporation reaction. In certain aspects, incorporation of lutetium-177 into or with tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) to provide ¹⁷⁷Lu-DOTATATE is greater than 99 mole percent based on the molar equivalent of lutetium-177 used in the incorporation reaction.
Preferably, an acidic aqueous formulation of lutetium-177 is admixed with the tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE). In a certain embodiment, a hydrogen halide or acid halide aqueous formulation of lutetium-177 is admixed with tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE).
Preferred preparations of ¹⁷⁷Lu-DOTATATE may include one or more and preferably each of the following steps:
1. Provide lutetium-177 in a vial or other container that can serve as a reaction vessel. The lutetium-177 suitably may be present in an aqueous acidic formulation, for example, an HCl formulation.
2. Admix DOTATATE peptide with an aqueous buffer composition (Reaction Buffer) that contains one or more ascorbate compounds (e.g., sodium ascorbate) and optionally, one or more gentisate compound (e.g., gentisic acid).
3. Admix the DOTATATE peptide from step 2 with the lutetium-177 composition of step 1. For example, the DOTATATE peptide composition from step 2 can be added to the vial that contains the lutetium-177.
4. The admixture of DOTATATE peptide and lutetium-177 then can be heated preferably with agitation, for example shaking with heating at 70-90° C., more typically 75-85° C. for up to 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 minutes.
5. After completion of the heating treatment, the admixture from step 4 above is allowed to cool for about 0.1, 0.25, 0.5, 1, 2, 3, 4, 5, 6 or 7 minutes at or below room temperature (e.g. 25° C.). Cooling may be facilitated by adding an aqueous composition (Formulation Composition) containing one or more ascorbate compounds (e.g. sodium ascorbate) and optionally, one or more gentisate compounds (e.g., gentisic acid) to the vial containing the admixture of step 4. The Formulation Composition added to the admixture may be at or below room temperature (e.g. 25° C.).
6. The admixture from step 5 then may be transferred to a vessel containing an aqueous composition (may be the same Formulation Composition as used in step 5) that comprises one or more ascorbate compounds and, in certain systems, one or more gentisate compounds. The mixture may be sterile filtered such as through one or more 0.22 μm filter and transferred to a container such as a syringe, vial or IV bag. Desired dosages can be dispensed for administration to a patient preferably within or example 5, 4, 3, 2, 1 or 0.5 days from completion of step 6.
In certain aspects, the preparation of ¹⁷⁷Lu-DOTATATE includes the exemplary process shown in FIG. 1 . For example, ¹⁷⁷Lu vial may contain ¹⁷⁷LuCl₃in 0.04 N HCl. A specifically preferred Reaction Buffer may suitably include 19 mg/mL gentisic acid, 57.3 mg/mL sodium ascorbate, and 3.7 mg/mL NaOH. A specifically preferred Formulation Composition may include 18.7 mg/mL gentisic acid, 56.1 mg/mL sodium ascorbate, 5.3 mg/mL NaOH, and 0.4 mg/mL diethylenetriaminepentaacetic acid (DTPA).
Other compositions shown in Examples are incorporated herein.

Pharmaceutical Compositions

In a further aspect, pharmaceutical compositions are provided.
Preferred pharmaceutical compositions may include an aqueous composition including 1) a complex of lutetium-177 and DOTATATE and 2) one or more ascorbate compounds.
Additional preferred pharmaceutical compositions may include an aqueous composition including 1) a complex of lutetium-177 and DOTATATE and 2) one or more gentisate compounds.
Still further preferred pharmaceutical compositions may include an aqueous composition including 1) a complex of lutetium-177 and DOTATATE; 2) one or more gentisate compounds; and 3) one or more ascorbate compounds.
Yet further preferred pharmaceutical compositions may include an aqueous composition including 1) a complex of lutetium-177 and DOTATATE; 2) one or more gentisate compound; 3) one or more ascorbate compounds; and 4) one or more sequestering agents.
Preferably, the radiochemical purity of a pharmaceutical composition is at least 95% where the composition is maintained at 25° C. or less and for 3, 4, or 5 days or more following preparation of the composition.
In certain preferred aspects, the pharmaceutical composition is free of unchelated lutetium-177 in an amount of not more than 2, 1.5, 1.0 or 0.5 weight % based on total weight of the pharmaceutical composition, such as may be determined by radiometric detection (including HPLC radiometric detection), where the composition is maintained at 25° C. or less and such purity levels are exhibited for 3 days or more following preparation of the composition.
In additional preferred aspects, the pharmaceutical composition is free of radiochemical impurities in an amount of not more than 5, 4, 3.5, 3, 2.5, 2, 1.5, 1 or 0.5 weight % based on total weight of the pharmaceutical composition, such as may be determined by radiometric detection (including HPLC radiometric detection), where the pharmaceutical composition is maintained at 25° C. or less and such purity levels of the pharmaceutical composition are exhibited for 3 days or more following preparation of the pharmaceutical composition.
In yet still additional preferred aspects, the pharmaceutical composition is free of chemical impurities in an amount of not more than 5, 4, 3, 2, 1 or 0.5 weight % based on total weight of the pharmaceutical composition, such as may be determined by HPLC/UV analysis, where the composition is maintained at 25° C. or less and such purity levels are exhibited for 3, 4 or 5 days or more following preparation of the pharmaceutical composition.
In yet still additional preferred aspects, the pharmaceutical composition is 1) free of unchelated lutetium-177 in an amount of not more than 2, 1.5, 1.0 or 0.5 weight % (such as may be determined by radiometric detection (including HPLC radiometric detection)); 2) free of radiochemical impurities in an amount of not more than 5, 4, 3.5, 3, 2.5, 2, 1.5, 1 or 0.5 weight % (such as may be determined by radiometric detection (including HPLC radiometric detection); and 3) free of chemical impurities in an amount of not more than 5, 4, 3, 2, 1 or 0.5 weight % (such as may be determined by HPLC/UV analysis), with all weight % based on total weight of the pharmaceutical composition, and where the pharmaceutical composition is maintained at 25° C. or less and such purity levels are exhibited for 3 days or more following preparation of the pharmaceutical composition.
In certain preferred aspects, purity levels, including radiochemical purities as referred to herein are determined by HPLC analysis, including radio-HPLC.
In certain aspects, the pharmaceutical composition is formulated for parenteral administration, such as intravenous, intramuscular, intradermal, subcutaneous, intrathecal or intraperitoneal administration. For example, the pharmaceutical composition is formulated for intravenous, intramuscular, subcutaneous or intradermal injection. In preferred aspects, the pharmaceutical composition is formulated for intravenous administration. In typical aspects, the pharmaceutical composition may be administered in a form of a pharmaceutical aqueous solution.
In certain aspects, the pharmaceutical composition is an aqueous solution, dispersion or other admixture such as for injection and comprises ¹⁷⁷Lu-DOTATATE and preferably 1) one or more ascorbate compounds; and/or 2) one or more gentisate compounds.
In certain preferred aspects, a pharmaceutical aqueous solution, dispersion or admixture is provided that includes: 1) a complex of lutetium-177 and DOTATATE; and 2) at least one stabilizer compound that preferably can inhibit radiolytic degradation of the composition during storage following preparation of the complex. ¹⁷⁷Lu-DOTATATE is suitably present in a concentration that it provides a volumetric radioactivity of at least 100 MBq/mL, preferably of at least 250 MBq/mL, or at least 300 or 400 MBq/mL within 1, 2, 3 or 4 days following preparation. In certain aspects, ¹⁷⁷Lu-DOTATATE is present in a concentration that it provides a volumetric radioactivity of from 100 to 1000 MBq/mL, preferably from 250 to 800 MBq/mL within 1, 2, 3 or 4 days following preparation.
In certain aspects, the one or more stabilizer compounds may be present in a total concentration of at least 5 mg/mL, preferably at least 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100 mg/mL of an aqueous pharmaceutical composition.
In certain aspects, the one or more stabilizer compounds may be present in a total concentration of at least 10 mg/mL, or at least 20, 30, 40, 50, 60 or 70 mg/mL of an aqueous pharmaceutical composition.
In additional certain aspects, the one or more stabilizer compounds may be present in a total concentration of at least 20 or 25 mg/mL, or at least 30, 40, 50, 60 or 70 mg/mL of an aqueous pharmaceutical composition.
In certain aspects, the one or more stabilizer compounds are one or more of gentisic acid (2,5-dihydroxybenzoic acid) or salts thereof, ascorbic acid (L-ascorbic acid) or salts thereof (e.g. sodium ascorbate), methionine, N-acetyl-L-methionine, N-acetyl-L-cysteine, histidine, melatonin, ethanol, or Se-methionine, preferably ascorbic acid or salts thereof.
In certain aspects, the one or more stabilizer compounds are one or more of 1) gentisic acid (2,5-dihydroxybenzoic acid) or salts thereof or 2) ascorbic acid (L-ascorbic acid) or salts thereof (e.g. sodium ascorbate).
In certain aspects, the pharmaceutical aqueous formulation has a shelf life of at least 24 hours at about 25° C. or less, at least 48 hours at about 25° C. or less, at least 72 hours at about 25° C. or less, or from 24 hours to 120 hours at about 25° C. or less, from 24 hours to 96 hours at about 25° C. or less, from 24 hours to 84 hours at about 25° C. or less, from 24 hours to 72 hours at about 25° C. or less, in particular a shelf life of 72 hours at about 25° C. or less.
In certain aspects, one, two or three total distinct stabilizer compounds are present during the complex formation of lutetium-177 and DOTATATE, preferably in an amount to result in a concentration of from 5 mg/mL or more of the one, two or three or more stabilizer compounds. As discussed, preferably at least one of the stabilizer compounds will be an ascorbate compound.
In certain aspects, as discussed, one or more stabilizer compounds may be added after formation of the complex of lutetium-177 and DOTATATE, for example upon completion of heating of an admixture of lutetium-177 and DOTATATE. As discussed, preferably at least one of the stabilizer compounds added after formation of ¹⁷⁷Lu-DOTATATE will be an ascorbate compound, for example where such stabilizer compound(s) are added upon temperature reduction at the conclusion of a heating step. In certain aspects, a gentisate compound also will be added after formation of ¹⁷⁷Lu DOTATATE, for example where such stabilizer compound(s) are added upon temperature reduction at the conclusion of a heating step. In that regard, the addition of an aqueous formulation containing one or more stabilizers promptly after heating is terminated can act to cool the ¹⁷⁷Lu DOTATATE reaction mixture.
In certain aspects, a pharmaceutical aqueous solution may further include a sequestering agent, for example added after formation of a complex of lutetium-177 and DOTATATE, suitably to remove uncomplexed lutetium-177. Suitable sequestering agents may include for example one or more aminopolycarboxylic acids, e.g. ethylenediaminetetraacetic acid (EDTA) and diethylenetriaminepentaacetic acid (DTPA) or a salt thereof, suitably in an amount to result in a concentration of from 0.01 to 0.50 mg/mL of the aqueous ¹⁷⁷Lu-DOTATATE composition.
In a particularly preferred aspect, ¹⁷⁷Lu-DOTATATE is provided as a sterile solution for intravenous use. A single-dose vial suitably will contain a dose of from 3.6±10% GBq to 11.1±10% GBq ¹⁷⁷Lu-DOTATATE. Four treatment cycles, represented by one injection per treatment cycle results in a cumulative dose of from 14.4±10% GBq to 44.4±10% GBq.
Preferably, the pH range of the ¹⁷⁷Lu-DOTATATE solution is 4.5 to 8.5. In a particularly preferred aspect, the pH range of the ¹⁷⁷Lu-DOTATATE solution is 5 to 7.

Methods of Treatment

As discussed, use of ¹⁷⁷Lu-DOTATATE (including 2 above) is provided to treat cancers, including cancers originated from foregut (e.g., stomach or duodenum), hindgut (e.g., left colon or rectum), midgut (e.g., jejunum, ileum, right colon, or appendiceal), lung, pancreas, and other organs (e.g., ovary, medulla, adrenal medulla, adrenal (pheochromocytoma), kidney, pituitary, thyroid or paraganglia).
In particular, ¹⁷⁷Lu-DOTATATE (including 2 above) may be used to treat neuroendocrine tumors, including to reduce neuroendocrine tumor size.
In such methods, ¹⁷⁷Lu-DOTATATE (including 2 above) can be administered to a subject such as a human in an amount effective to treat the cancer (e.g., reduction of tumor size), such as at a dose of about 1 or 2 GBq to about 15 GBq or 20 GBq or more per treatment cycle, and in certain aspects from 3.6 GBq to about 11.1 GBq per treatment cycle, and can be suitably administered from a unit dose in a vial or a syringe or as a bulk solution in a vial or a syringe prepared from a cold-kit prepared with lutetium-177 at a local or central radiopharmacy or through cGMP central manufacturing. Total dose administered for therapy including 4 treatment cycles is about 14.4 GBq to about 40.8 GBq or 44.4 GBq.
The effective amount of the ¹⁷⁷Lu-DOTATATE radiopharmaceutical administered to a patient will generally be determined by considering the patient record. However, the effective amount suitably may be within a range of about at a dose of about 1 or 2 GBq to about 15 GBq or 20 GBq or more per dose, more typically about 3.6 GBq to 11.1 GBq per dose, for example, about 3.6, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.4, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.1 GBq per dose, or any range between two of the above values. The dose can be administered from a unit dose in a vial or a syringe or as a bulk solution in a vial or a syringe prepared from a cold-kit prepared with lutetium-177 at a local or central radiopharmacy or through cGMP central manufacturing.
If necessary or desirable, the treatment may involve more than one administration of an effective amount of ¹⁷⁷Lu-DOTATATE. It is generally beneficial to repeat the administration of ¹⁷⁷Lu-DOTATATE to the subject after 7 to 56 days, such as at a 4 to 8 week interval.
In a particularly preferred protocol, the ¹⁷⁷Lu-DOTATATE dosage form is a sterile aqueous solution that is administered by intravenous injection. The dosing regimen may include multiple infusions such as 4 infusions at effective dosages such as from 3.6 GBq±10% to 11.1 GBq±10% each, administered about 4, 5, 6, 7 or 8 weeks apart.
In certain aspects, the methods include assessing neuroendocrine tumors of a patient to be somatostatin receptor positive, for example with ⁶⁸Ga-DOTATATE and a positron emission tomography scan.

Personalized Dosing

In further aspects, dosing protocols may be utilized to provide dosing amounts for a specified patient based on one or more of the patient's characteristics.
In particular, ¹⁷⁷Lu-DOTATATE may be administered in dosage amounts based on dosimetry assessments. For example, following administration of ¹⁷⁷Lu-DOTATATE, the patient may be assessed by SPECT (e.g. 3D SPECT-CT imaging) and planar scans or other analysis to allow individualized dosimetry. Multiple scans may be performed, for example at approximately 4, 24 and 72 hours following dosing, or at a single time point or other schedule. The scans can be used to determine doses absorbed such as by tumors, kidneys and bone marrow of the patient. Based on that assessment, additional dosing of the patient can be modified, in particular either increased or decreased to enhance efficacy or safety.
Additionally, a patient's glomerular filtration rate (GFR) also may be assessed typically by a determination of creatinine (Cr) clearance.
Preferably, a patient receiving ¹⁷⁷Lu-DOTATATE will be assessed both by dosimetry and Cr clearance/GFR. By such a combined assessment, patients may be dosed with ¹⁷⁷Lu-DOTATATE at a GFR of 40 mL/min and as low as 30 mL/min.

Combination Therapy

¹⁷⁷Lu-DOTATATE (including 2 above) suitably may be administered to a subject in conjunction or combination with one or more other therapeutic agents, particularly one or more other chemotherapeutic agents.
In one aspect, a subject may receive treatment with ¹⁷⁷Lu-DOTATATE in combination with a regime of one or more somatostatin compounds, such as octreotide (e.g. Sandostatin) or lanreotide (e.g. Somatuline Autogel).
A subject also may receive treatment with ¹⁷⁷Lu-DOTATATE in combination with a regime of one or more anticancer agents, for example capecitabine, temozolomide, steptozotocin, 5-fluroouracil, cisplatin, carboplatin, etoposide, and/or doxorubicin.
As used herein, the term “in combination” in the context of the administration of a therapy to a subject refers to the use of more than one therapy for therapeutic benefit. The term “in combination” in the context of the administration can also refer to the prophylactic use of a therapy to a subject when used with at least one additional therapy. The use of the term “in combination” does not restrict the order in which the therapies (e.g., a first and second therapy) are administered to a subject. A therapy can be administered prior to (e.g., 1 minute, 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks before), concomitantly with, or subsequent to (e.g., 1 minute, 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks after) the administration of a second therapy to a subject in need of treatment as disclosed herein. The therapies are administered to a subject in a sequence and within a time interval such that the therapies can act together. In a particular embodiment, the therapies are administered to a subject in a sequence and within a time interval such that they provide an increased benefit than if they were administered otherwise. Any additional therapy can be administered in any order with the other additional therapy.

Packaged ¹⁷⁷Lu-DOTATATE and Kits

As discussed above, treatment kits are also provided, including cold kits where the ¹⁷⁷Lu-DOTATATE can be prepared shortly before administration such as in a medical facility, for example a hospital laboratory or radiopharmacy. In such a kit, DOTATATE may be provided in a vial or other container in lyophilized or other form separate from lutetium-177. The DOTATATE and lutetium-177 are reacted as disclosed herein at the medical facility to provide ¹⁷⁷Lu-DOTATATE which then can be promptly administered to a patient.
The kit may be a single dosage kit including ¹⁷⁷Lu-DOTATATE or pharmaceutical composition as described herein. Alternatively, the kit may be a multiple dosage kit comprising ¹⁷⁷Lu-DOTATATE or pharmaceutical composition as described herein.
In a further aspect, packaged preparations or products of ¹⁷⁷Lu-DOTATATE are also provided. A packaged preparation may comprise 1) ¹⁷⁷Lu-DOTATATE and optionally 2) instructions for using ¹⁷⁷Lu-DOTATATE for treating a cancer such as a neuroendocrine tumor. Preferably, the packaged preparation will comprise a therapeutically effective amount of ¹⁷⁷Lu-DOTATATE.
In certain exemplary packaged preparations or products, ¹⁷⁷Lu-DOTATATE can be packaged in suitable containers labeled, for example, for use as a therapy to treat a subject suffering from cancer e.g. a neuroendocrine tumor. The containers can include ¹⁷⁷Lu-DOTATATE and one or more of a suitable stabilizer compounds as disclosed herein. A product can include a container (e.g., a vial or the like) containing ¹⁷⁷Lu-DOTATATE. In addition, an article of manufacture or kit further may include, for example, packaging materials, instructions for use, syringes, delivery devices, for treating the targeted condition, such as a neuroendocrine tumor or other cancer.
A packaged system or product may also include a legend (e.g., a printed label or insert or other medium (e.g., an audio or video file) describing the product's use). The legend can be associated with the container (e.g., affixed to the container) and can describe the manner in which the compositions therein should be administered (e.g., the frequency and route of administration), indications therefor, and other uses. The compositions can be ready for administration (e.g., present in dose-appropriate units), and may include one or more additional pharmaceutically acceptable adjuvants, carriers or other diluents.
The following non-limiting examples are illustrative.

Example 1: ¹⁷⁷Lu-DOTATATE Manufacturing

Table 1 describes all input materials into the manufacturing process

TABLE 1

Ingredient	Quantity per batch

¹⁷⁷LuCl₃(in 0.04N HCl)	≤88.4 GBq¹
DOTATATE peptide	0.2-0.8 mg²

0.04N HCl	1N HCl	<1 mL
solution	Sterile Water for Injection, USP

Reaction Buffer³

(19 mg/mL gentisic acid, 57.3 mg/mL sodium ascorbate,

3.7 mg/mL NaOH)

Gentisic acid	0.6-2.4 mL, particularly
Sodium ascorbate, USP/EP	1.2 mL
Sterile Water for Injection, USP
2.0 N NaOH

Formulation Buffer

(18.7 mg/mL gentisic acid, 56.1 mg/mL sodium ascorbate,

5.3 mg/mL NaOH, 0.4 mg/mL DTP A)

Reaction buffer	6.95-115.6 mL
2.0N NaOH
Diethylenetriamine pentaacetic (DTPA) acid

¹At start of synthesis
²Depending on number of doses/batch
³Also a component of the Formulation Buffer

Table 2 below describes the ingredients found in the final formulated drug product.

TABLE 2

Ingredient	Quantity^{1, 2}	Role in formulation

¹⁷⁷Lu-DOTATATE	~80.3	GBq³	Drag substance
	≤0.96	GBq/mL³
Gentisic acid	16-19	mg/mL	Radioprotectant and buffer
Sodium ascorbate,	49-56	mg/mL	Radioprotectant and buffer
USP/EP
Sterile Water for	15.5-118	mL	Diluent
Injection, USP
DTPA	0.3-0.4	mg/mL	Chelator of free [Lu-177]
			to minimize physiological
			uptake
Sodium Hydroxide,	4.6-5.1	mg/mL	Adjustment of buffer pH
USP-NF

¹Total batch volume = 16-120 mL
²Ranges depend on batch size and volume required to yield target radioactivity concentration
³At time of calibration (TOC), defined as 07:00 ET on day of manufacture

Fabrication Process

Step 1 described in this section takes place in a Grade A biological safety cabinet. All subsequent steps described in this section take place within a Grade A shielded isolator. All solutions, including Reaction Buffer and Formulation Buffer, are prepared in a Grade C clean room. In summary, in the incorporation reaction, the DOTATATE peptide is mixed with reaction buffer and ¹⁷⁷LuCl₃, followed by heating and shaking to form the drug complex substance.
The heater/shaker undergoes installation, operation and performance qualification prior to use. Qualification has confirmed that the required temperature and shaking conditions for the incorporation reaction (radiolabeling) can be maintained for the specified time period, as described in Step 4 below.
The amount of DOTATATE peptide used (Step 2) is based on the total amount of ¹⁷⁷Lu-DOTATATE activity (itself based on the number and timing of patient doses) and the volume required to yield a radioactive concentration of <0.96 GBq/mL at time of calibration (TOC). The TOC is defined as 07:00 ET on the day of manufacture (DOM).

- 1. Depending on the amount of ¹⁷⁷Lu starting activity required, the appropriate volume of 0.04 N HCl is added to the supplier vial containing ¹⁷⁷LuCl₃in 0.04 N HCl to bring the total volume to
- a. 1000 μL (if 160-320 μs of DOTATATE peptide are used)
- b. 2000 μL (if 480-640 μs of DOTATATE peptide are used)
- 2. 1-4×750 μL volume(s) of DOTATATE peptide are prepared in reaction buffer. The solution is gently agitated to ensure complete dissolution of the peptide. Each 750 μL volume of DOTATATE peptide solution contains 200 μg DOTATATE.
- 3. If only 1× or 3×750 μL aliquots of DOTATATE solution are required, an additional 1×600 μL volume of reaction buffer is prepared.
- 4. The reaction solution is prepared by adding 600 μL volumes of DOTATATE solution from Step 2 (and if applicable reaction buffer from Step 3) to the supplier vial from Step 1 (containing diluted ¹⁷⁷LuCl₃). The total reaction solution volume is 2.2 mL (160-320 μg of peptide) or 4.4 mL (480-640 μg of peptide).
- 5. Using the qualified and pre-heated heater/shaker, the reaction solution is heated at 80±5° C. for 15±1 min with shaking at 350 rpm.
- 6. The reaction solution vial containing the drug substance is allowed to cool at room temperature for at least 5 min.

Subsequent steps consist of transferring the drug substance (in reaction buffer) through a tandem 2×0.22 μm sterile filter chain, to a 100 mL sterile bulk vial containing formulation buffer, and aseptic dispensing of the final drug product into the final container closure. These steps are accomplished using a dispensing apparatus consisting of a series of stopcocks to which various tubing, syringes, vials and needles are attached to facilitate transfer and dispensing of the drug product. Steps 7-10 take place within a Grade A shielded isolator unit.

- 7. The ¹⁷⁷Lu-DOTATATE drug substance is immediately transferred from the reaction vial, through a tandem 2×0.22 μm sterile filter chain, to a 100 mL bulk vial that contains (post-reaction vial rinsing) 6.95-115.6 mL of formulation buffer. This step forms the drug product and represents end of synthesis (EOS).
- 8. Samples are removed from this vial for sterility and QC testing.
- 9. Patient doses are dispensed into final 30 mL container closures.

The total synthesis time, from Step 3 to Step 7 described above, is approximately 120 min.
FIG. 2 shows a flow chart for a preferred exemplary fabrication process.

Example 2: Treatment Protocol

A human patient with neuroendocrine tumors is selected for treatment after their tumors have been shown to be somatostatin receptor positive with ⁶⁸Ga-DOTATATE and a positron emission tomography scan.
¹⁷⁷Lu-DOTATATE in a sterile aqueous solution as set forth in Table 2 of Example 1 above is administered by intravenous injection to the patient. Single-dose vials are used that contain from 3.6±10% GBq to 11.1±10% GBq ¹⁷⁷Lu-DOTATATE. The ¹⁷⁷Lu-DOTATATE is administered over four treatment cycles, with one injection per treatment cycle that provides a cumulative dose of from 14.4 GBq to 44.4 GBq.

Example 3

Materials and Methods

An Eppendorf Thermomixer C, 96-well Eppendorf Microplates and ThermoFisher 96-well plate sealing tape were used for small scale reaction screening. A VWR heating block was used for scaled up reactions. A Waters Acquity Arc UHPLC with a radioactivity detector and an Acquity QDa mass detector were used for analysis. A Thermo Scientific Acclaim 120 C18 column (150×3 mm, 120 Å, 3 μm) was used for chromatographic separation. An Eckert & Ziegler AR-2000 Radio-TLC Imaging Scanner was used for radio-iTLC analysis.
The ^natLu-DOTATATE reference standard was purchased from ABX advanced biochemical compounds GmbH (Radeberg Germany) and the DOTATATE precursor was obtained from Auspep Clinical Peptides.

Radio-iTLC Method

Agilent iTLC-SA chromatography paper was used for radio-iTLC analysis. The mobile phase used for chromatographic separation was 20% CH₃OH in 0.1M citrate buffer. The sample volume spotted on the paper was 1 μL. The migration distance was 9 cm.

Instrument Settings


Column temperature	40° C.
Mobile phase	A: H₂O + 0.1% TFA
	B: CH₃CN + 0.1% TFA
System wash & seal/needle wash	50% CHCN/H₂O
Flow rate	0.6 mL/minute
Autosampler tray temperature	Ambient temperature
UV Detector parameters	Single λ = 220 nm
	Sampling Rate: 1 point/sec
Radiometric Detector parameters	eSatin: 60 Hz, Sampling Rate:
	2 points/second, Scale Factor 1000.
	Bioscan FC: Detector PMT,
	Integration Time 1 sec.
	ERM: 12 CPM

HPLC Mobile Phase Gradient


Time	% A	% B	Gradient Curve

0	85	15	6 (Linear)
17.0	68	32	6 (Linear)
17.5	40	60	6 (Linear)
20.5	40	60	6 (Linear)
21.0	85	15	Re-equilibration
28.0	85	15

Buffer Development

The volume of 1M NaOH or 1M HCl to reach the desired pH in the incorporation reaction admixture or the final drug complex product mixture was determined experimentally.

Incorporation Reaction Condition Screening Experiment

Non-Variable Parameters

For the incorporation reaction screening experiments, the following components were added to each reaction well:


		Volume
	Component	(μL)

	^175/177LuCl₃in 0.04N HCl	45.5
	Reaction buffer (see below) + precursor	54.5
	peptide (DOTATATE)

The peptide concentration in the reaction mixture was 90.9 μg/mL. The radioactive concentration (RAC) of each incorporation (radiolabelling) reaction was 1 μCi/mL at the start of the experiment. ¹⁷⁵LuCl₃was spiked into the reaction to reach the Lu/precursor molar ratio in a comparable manufacturing process with ¹⁷⁷LuCl₃at a specific activity of 3.8 GBq/μg, using a 7.8 GBq input activity.

Variable Parameters

Screening of incorporation reaction conditions was performed with the following reaction variables of the Reaction buffer as referred to above and used in 54.5μ:


	Reaction buffer components	Value

	Sodium ascorbate (mg/mL)**	100, 50, 10
	Gentisic acid (mg/mL)**	15, 7, 0

	Reaction buffer and
	incorporation reaction
	parameters	Value

	pH*	4, 5, 6
	Incorporation reaction	45, 60
	temperature (° C.)
	Incorporation reaction time	10, 15, 30
	(minutes)

	*pH specified as tested in final incorporation reaction (radiolabeling) mixture.
	**Sodium ascorbate and gentisic acid concentrations specified for incorporation reaction buffer (before addition of ^175/177LuCl₃in 0.04N HCl).

Variable Parameters for the Second Round of Incorporation Reaction Condition Screening

The second round of incorporation reaction condition screening was performed with the following reaction variables. Not all permutations of these variables were tested.


	Reaction parameter	Value

	pH*	4.5, 5
	Sodium ascorbate (mg/mL)**	100, 75
	Gentisic acid (mg/mL)**	15, 7, 0
	Reaction temperature (° C.)	45, 60
	Reaction time (minutes)	30, 60

	*pH specified as tested in final incorporation reaction (radiolabeling) mixture.
	**Sodium ascorbate and gentisic acid concentrations specified for incorporation reaction (radiolabeling) buffer (before addition of ^175/177LuCl₃in 0.04N HCl).

Evaluation of Incorporation Reaction Condition Screening Experiments

For the initial incorporation reaction condition screening experiment, each experiment was evaluated by radio-iTLC immediately after the reaction. The various time-points were sampled from the same reaction volume and spotted onto the iTLC paper to quench the reaction. For the second round of incorporation reaction experiments, radio-iTLC and radio-HPLC analysis was performed.

Formulation Condition Screening Experiment

Incorporation Reaction

A single incorporation reaction was performed and aliquoted into different formulation buffers for the formulation condition screening experiment. The following parameters were used for the incorporation reaction:


	Incorporation reaction mixture component	Value

Precursor peptide (DOTATATE)	90.9	μg/mL
concentration*
Radioactive (¹⁷⁷Lu) concentration*	3.36	GBq/mL
Sodium ascorbate**	100	mg/mL
Gentisic acid**	0	mg/mL

	Incorporation reaction parameter	Value

	pH
	5

Reaction mixture volume	0.416	mL
Reaction temperature
60°	C.
Reaction time
30	minutes

*Parameter specified in final incorporation reaction mixture.
**Parameter specified for incorporation reaction (radiolabeling) buffer (before addition of ¹⁷⁷LuCl₃in 0.04N HCl to reach the final incorporation reaction mixture).

Non-Variable Formulation Parameters

For the formulation condition screening experiments, the following components were added to each reaction well:
Volume

Component (μL)

Incorporation reaction mixture 22.0

Formulation buffer 59.0

The following parameters were held constant for each formulation condition test:


	Parameter	Value

	DTPA*	0.29 mg/mL
	Radioactive concentration*	0.912 GBq/mL
	Reaction mixture dilution ratio	1:3.68 (Reaction mixture
		to final product volume)

	*Parameter specified in final product mixture.

Variable Formulation Parameters

Screening of formulation conditions (to provide ¹⁷⁷Lu-DOTATATE pharmaceutical compositions) was performed with the following reaction variables:


	Reaction parameter*	Value

	pH

	5, 6

	¹⁷⁷Lu-DOTATATE
	pharmaceutical composition
	components	Value

	Sodium ascorbate (mg/mL)	100, 75, 50
	Gentisic acid (mg/mL)	18, 11, 0

	*Parameter specified as tested in final product mixture

Evaluation of Formulation Screen Experiments

The formulation screen experiments were analyzed by radio-iTLC on day 0, 1, 3 and 5. The sample was analyzed by radio-HPLC on day 5.

Scale Up Experiments

Incorporation Reaction Parameters

Two incorporation reactions were performed. The following components were added to each reaction vessel:


		Volume
	Component	(μL)

	¹⁷⁷LuCl₃in 0.04N HCl	500
	Reaction buffer (see incorporation reaction #1	600
	(L1) and (L2) #2 values below) + 100 μg
	precursor peptide (DOTATATE)

Formulation Parameters

The following components parameters were used for the two incorporation reactions:


		Incorporation	Incorporation
		reaction #
1	reaction #2
	Component	(L1) Value	(L2) Value

	Radioactive concentration	3.36 GBq/mL	3.36 GBq/mL
	Sodium ascorbate*	100 mg/mL	100 mg/mL
	Gentisic acid*	0 mg/mL

		Incorporation	Incorporation
		reaction #
1	reaction #2
	Parameter	(L1) Value	(L2) Value

	pH
	5	5
	Reaction time	30 minutes	60 minutes
	Reaction temperature
	60° C.	45° C.

	*Parameter specified for incorporation reaction buffer (before addition of ¹⁷⁷LuCl₃in 0.04N HCl to reach the final incorporation reaction mixture). 100 mg/mL sodium ascorbate in the buffer corresponds to 54 mg/mL in the incorporation reaction.

Two different formulation conditions were used for each of the two incorporation reactions (L1 and L2). The following components were combined to obtain four different final products:


		Volume
	Component	(μL)

	Incorporation reaction mixture (L1 and L2)	500
	Formulation buffer (F1 and F2)	1360

The following parameters were used for the two different product formulations:


		Formulation #1	Formulation #2
	Component	(F1) Value	(F2) Value

	Radioactive concentration	0.913 GBq/mL	0.913 GBq/mL
	Sodium ascorbate*	75 mg/mL	50 mg/mL
	Gentisic acid*	15 mg/mL	15 g/mL

		Formulation #
1	Formulation #2
	Parameter	(F1) Value	(F2) Value

	pH
	6	6
	DTPA	0.29 mg/mL	0.29 mg/mL
	Reaction mixture dilution	1:3.72	1:3.72

	ratio

	*Parameter specified for radiolabeling buffer (before addition of ¹⁷⁷LuCl₃in 0.04N HCl to reach the final incorporation reaction mixture)

Evaluation of Scale Up Experiments

Each of the two incorporation reaction mixtures were formulated with the two formulation specifications to generate four samples, L1F1, L2F2, L2F1, and L2F2. The four products were tested by radio-iTLC and radio-HPLC on day 0, 1, 2 and 5. The products were stored at room temperature over the course of the 5-day study.

Results and Discussion

Incorporation Reaction Optimization

Incorporation reaction screening was conducted to evaluate the effect of pH, sodium ascorbate concentration, gentisic acid concentration, reaction temperature and reaction time on radiometal (¹⁷⁷Lu) incorporation (Table 3).

TABLE 3

Incorporation condition screening variables.

	Incorporation reaction
	component	Value

	Sodium ascorbate (mg/mL)	100, 50, 10
	Gentisic acid (mg/mL)	15, 7, 0

	Reaction parameter	Value

	pH


	4, 5, 6
	Reaction temperature (° C.)	45, 60
	Reaction time (minutes)	10, 15, 30

Optimal Incorporation Reaction Achieved at

pH

5, 30 Min Reaction Time, and 60° C. Reaction Temperature

Analysis of the reactions by radio-iTLC indicated that optimal incorporation reaction (radiolabeling) was achieved at pH 5. The reaction was worse at pH 4 and did not notably proceed at pH 6 (FIG. 3 ). Performing the reaction at higher temperature and for a longer duration allowed for more efficient incorporation of Lu-177.

Sodium Ascorbate Improves Incorporation Reaction

The presence of sodium ascorbate had a significant beneficial effect on radiometal (¹⁷⁷Lu) incorporation in the test reactions. The reactions with 100 mg/mL of sodium ascorbate in the reaction buffer (corresponding to 54 mg/mL in the radiolabeling mixture) had the highest % ¹⁷⁷Lu incorporation as measured by radio-iTLC. Gentisic acid was well tolerated in the reaction at all concentrations tested. See results set forth in FIG. 4 .

Second Round of Incorporation Reaction (Radiolabeling) Screening Experiments

Eight incorporation reactions were performed and evaluated by radio-iTLC and radio-HPLC (Table 4 below).

TABLE 4

Incorporation reaction conditions for reactions tested in second round of radiolabeling
screening experiments.

				Sodium	Gentisic	Radio-iTLC (%
Sample		Reaction temp	Reaction time	ascorbate	acid	incorporation ±
#	pH	(° C.)	(min)	(mg/mL)	(mg/mL)	SD)

1	5	60	30	100	15	96.9 ± 0.4
2	5	60	30	100	7	99.1 ± 0.1
3	5	60	30	100	0	98.9 ± 0.1
4	4.5	60	30	100	7	98.8 ± 0.2
5	5	60	30	75	7	98.9 ± 0.1
6	4.5	60	30	100	0	96.1 ± 4.2
7	5	60	30	75	0	99.0 ± 0.2
8	5	45	60	100	7	98.1 ± 0.9

For all samples, the only radioactive peak that was observed was ¹⁷⁷Lu-DOTATATE (FIG. 5 ), although due to the trace amount of activity used in the reaction (1 μCi/mL), the radiation detector signal was very low. Significant tailing of the product peak was also observed with an elevating baseline after each injection, which prevented the injection of more material per sample injection. This problem carried into subsequent experiments and was ultimately resolved by replacing the fluid path line (Waters) in the radiation detector.
Comparison of the UV trace between samples #1, #2 and #3 indicated that increased chemical impurities are detected in samples containing more gentisic acid (see FIG. 6A). Analysis of gentisic acid by HPLC indicated that some of these peaks are impurities that are present in the gentisic acid material as received. Comparison of the UV traces between samples #2 and #8 show increased amounts of chemical impurities when a higher reaction temperature is used. Control experiments showed that new impurities form from gentisic acid and/or its initial impurities when heated in solution.
Due to the increase chemical purity with higher concentrations of sodium ascorbate (100 mg/mL) and lower concentrations of gentisic acid (0 mg/mL), sample #3 labeling conditions was selected for further investigation in formulation screening tests.

Formulation Evaluation

Formulation screening was conducted to evaluate the effect of pH, sodium ascorbate concentration and gentisic acid concentration on the stability of ¹⁷⁷Lu-DOTATATE. Each sample was tested by radio-iTLC on days 0, 1, 3 and 5 (Tables 5-7 below).

TABLE 5

Formulation screening iTLC results for samples with 100 mg/mL of sodium ascorbate.

Sodium ascorbate-100 mg/mL

Gentisic acid

Radio-iTLC % Lu labeling

pH	(mg/mL)	Day 0	Day 1	Day 3	Day 5

5	25	97.9	98.2	95.6*	20.7*
5	15	97.4	98	99.1*	98.4*
5	0	99	97.5	98.8	97.5
6	25	98.8	98.3	98.7	98
6	15	98.4	98.6	98.5	97.1
6	0	98.6	98.7	98.7	98.7

*On Day 3, limited buffer observed in wells A1, A2, B1. Possibly due to evaporation from incorrect sealing of plate in top left corner.

TABLE 6

Formulation screening iTLC results for samples with 75 mg/mL of sodium ascorbate.

Sodium ascorbate-75 mg/mL

Gentisic acid

Radio-iTLC % Lu labeling

pH	(mg/mL)	Day 0	Day 1	Day 3	Day 5

5	25	96.9	98.2	97.5*	98.4*
5	15	99.1	98.2	98.1	97.9
5	0	98.1	98	98.1	97.4
6	25	98.6	98.2	98.4	98.1
6	15	98.7	98.2	98.9	97.9
6	0	98.5	98.3	99	98.2

*On Day 3, limited buffer observed in wells A1, A2, B1.

TABLE 7

Formulation screening iTLC results for samples with 50 mg/mL of sodium ascorbate.

Sodium ascorbate-50 mg/mL

Gentisic acid

Radio-iTLC % Lu labeling

pH	(mg/mL)	Day 0	Day 1	Day 3	Day 5

5	25	97.3	98.5	98.5	98.5
5	15	98.1	98.7	98	98
5	0	98.2	98.8	98.4	97.9
6	25	98.7	98.6	98.4	98.2
6	15	98.3	98.3	98.4	97.3
6	0	98.9	98.8	98.4	98.6

All samples were analyzed by radio-HPLC on day 5. Tailing of the product was observed which resulted in an elevated radiation detector baseline.
While the tailing and elevated baseline of the product peak complicated interpretation of the sample spectra, some general conclusions could be made from the experiment (FIG. 7 ). Notably, fewer impurities were present at day 5 when the product was at pH 6. Comparison of the UV detector spectra was completed but no major differences were observed. Two formulation conditions were selected for scale up testing: F1—pH 6, sodium ascorbate 75 mg/mL, gentisic acid 15 mg/mL, and F2—pH 6, sodium ascorbate 50 mg/mL, gentisic acid 15 mg/mL.

Scale Up and Stability Testing

Two labeling conditions and two formulation conditions to generate four different products were chosen for scale up and stability testing (Tables 8-7). All samples were analyzed by radio-iTLC (Tables 10-11) and radio-HPLC on day 0, 1, 2 and 5. Tailing of the product peak was not observed after replacing the fluid path to the radiation detector. All products showed a prominent peak at the expected retention time for ¹⁷⁷Lu-DOTATATE with several impurities eluting both before and after the product peak. These impurities were visualized by decreasing the scale on the y-axis (FIG. 7 ).

TABLE 8

Labeling conditions for scale up and stability test.

		Reaction	Reaction	Sodium	Gentisic
		temp	time	ascorbate	acid
Sample #	pH	(° C.)	(minutes)	(mg/mL)	(mg/mL)

L1	5	60	30	100	0
L2	5	45	60	100	0

TABLE 9

Formulation conditions for scale up and stability test.

		Sodium	Gentisic
		ascorbate	acid
Sample #	pH	(mg/mL)	(mg/mL)

F1	6	75	15
F2	6	50	15

TABLE 10

Radio-iTLC results for L1 scale up and stability tests.

		L1
		Radio-ITLC % Lu labeling

Sample	Day	0	Day 1	Day 2	Day 5

F1	98.2	98.9	98.1	98.8
F2	97.9	98.8	99.3	98.6

TABLE 11

Radio-iTLC results for L2 scale up and stability tests.

			L2
			Radio-ITLC % Lu labeling

	Sample	Day	0	Day 1	Day 2	Day 5

F1	96.7	97	97.9	96.3
F2	94.8	97.3	98.1	97.8

The above results show that the synthesis of ¹⁷⁷Lu-DOTATATE proceeds favorably at pH 5, and that using the highest incorporation reaction temperature tested (60° C.) for 30 minutes allowed for the highest degree of Lu-177 incorporation. The ascorbate compound concentration in the incorporation reaction mixture positively correlated with radiometal incorporation. The highest concentration tested (100 mg/mL in the reaction buffer=54 mg/mL in the radiolabeling mixture) led to the best results by radio-iTLC (see FIG. 4 ).
Formulation screening indicated that the product was stable at all conditions tested, but that a product at pH 6 results in fewer impurities than pH 5. Scale up and stability testing showed several impurities are formed in the labeling mixture and that while most of them are stable once the produce is formulated, some increase over 5 days stored at room temperature. In general, fewer impurities were observed when the product was labeled using a lower reaction temperature.

Example 4: Composition of Single Dose Batch of 177Lu-DOTATATE (97mCi/3.59 GBq Input Lu-177)

The following compositions are prepared in accordance with procedures set forth in Examples 1 and 3 above. For all solutions, 1 g/mL density is assumed. As generally referred to herein, unless otherwise indicated, a Reaction Buffer (or similar term) is used in an incorporation reaction. As generally referred to herein, unless otherwise indicated, a Formulation Buffer (or similar term) is used to prepare ¹⁷⁷Lu-DOTATATE pharmaceutical composition from incorporation reaction mixture.

Part 1:

Quantities

Ingredients	Amount	Unit	Conc.

2N NaOH (pH 12-13)

Sterile Water for Injection (SWFI)	93.30	ml	N/A
NaOH	7.60	g	81.46	mg/mL
			2.04	N

Reaction Buffer

Sterile Water for Injection (SWFI)	130.30	g	N/A
Sodium L-Ascorbate	7.67	g	56.15	mg/mL
Gentisic acid	2.57	g	18.81	mg/mL
2N NaOH	6.30	g	3.76	mg/mL

Formulation Buffer

Reaction buffer	120.90	g	N/A
DTPA	46.30	mg	0.38	mg/mL
2N NaOH	2.50	g	5.33	mg/mL
Sodium L-Ascorbate		g	55.01	mg/mL
Gentisic Acid		g	18.43	mg/mL

Part 2:


			mg/mL in
			Total
			Incorporation
			Reaction
Composition of Incorporation Reaction	volume	mg	Volume

160 ug Precursor (DOTATATE)	0.60 mL
dissolved in Reaction Buffer*
Amount Ascorbate in Reaction Buffer		33.69	21.06
Amount GentisicAcid in Reaction Buffer		11.29	7.06
Amount NaOH in Reaction Buffer		2.25	1.41
Volume of Lu-177 in 0.04M HCl	1.00 mL
Total Volume=	1.60 mL

*Reaction Buffer from Part 1

Part 3:


			mg/mL in
Composition of Pharmaceutical			Pharmaceutical
Composition Prepared from Part 2	volume	mg	Composition

Formulation Buffer*	6.95 mL
Amount Ascorbate in Formulation		382.33	44.72
Buffer
Amount Gentisic acid in Formulation		128.11	14.98
Buffer
Amount DTPA in Formulation Buffer		2.61	0.30
Amount NaOH in Formulation Buffer		37.05	4.33
Incorporation Reaction Solution**	1.60 mL
Amount Gentisic acid in Incorporation		11.29	1.32
Reaction Solution
Amount Ascorbate in Incorporation		33.69	3.94
Reaction Solution
Amount NaOH from in Incorporation		2.25	0.26
Reaction Solution
Total Volume=	8.55 mL

	Pharmaceutical Composition
	48.66 mg/mL total ascorbate
	16.30 mg/mL total gentisic acid
	0.30 mg/mL DTPA
	4.60 mg/mL NaOH

*From Part 1
**From Part 2

Example 5: Composition of Multi-Dose Batch of 177Lu-DOTATATE (2389.2 mCi/88.4 GBq Input Lu-177)

The following compositions are prepared in accordance with procedures set forth in Examples 1, 3 and 4 above. For all solutions, 1 g/mL density is assumed.

Part 1:

Quantities

Ingredients	Amount	Unit	Conc.

2N NaOH (pH 12-13)

Sterile Water for Injection (SWFI)	102.70	ml	N/A
NaOH	8.40	g	81.79	mg/mL
			2.04	N

Reaction Buffer

Sterile Water for Injection (SWFI)	129.70	g	N/A
Sodium L-Ascorbate	7.93	g	58.36	mg/mL
Gentisic acid	2.63	g	19.36	mg/mL
2N NaOH	6.18	g	3.72	mg/mL

Formulation Buffer

Reaction buffer	119.10	g	N/A
DTPA	48.00	mg	0.40	mg/mL
2N NaOH	2.30	g	5.20	mg/mL
Sodium L-Ascorbate		g	57.25	mg/mL
Gentisic Acid		g	18.99	mg/mL

Part 2:

640 ug Precursor (DOTATATE)	2.40 mL
dissolved in Reaction Buffer*
Amount Ascorbate in Reaction Buffer		140.06	30.38
Amount Gentisic Acid in Reaction		46.45	10.08
Buffer
Amount NaOH in Reaction Buffer		8.93	1.94
Volume of Lu-177 in 0.04M HCl	2.21 mL
Total Volume=	4.61 mL

*Reaction Buffer from Part 1

Part 3:

Formulation Buffer*	105.30 mL
Amount Ascorbate in Formulation		6028.91	54.85
Buffer
Amount Gentistic Acid in Formulation		1999.50	18.19
Buffer
Amount DTPA in Formulation Buffer		41.63	0.38
Amount NaOH in Formulation Buffer		547.47	4.98
NaOH
Incorporation Reaction Solution**	4.61 mL
Amount Gentisic acid in Incorporation		46.45	0.42
Reaction solution
Amount Ascorbate in Incorporation		140.06	1.27
Reaction solution
Amount NaOH in Incorporation		8.93	0.08
Reaction solution
Total Volume=	109.91 mL

	Pharmaceutical Composition
	56.13 mg/mL total ascorbate
	18.61 mg/mL total gentisic acid
	0.38 mg/mL DTPA
	5.06 mg/mL NaOH

*From Part 1
**From Part 2

Example 6: Additional Compositions

The following compositions are prepared in accordance with procedures set forth in Examples 1, 3, 4 and 5 above. For all solutions, 1 g/mL density is assumed.

Quantities

2N NaOH (pH 12-13)

Ingredients	Amount	Unit	Conc.

Sterile Water for Injection (SWFI)	98.00	ml	N/A
NaOH	8.00	g	81.63	mg/mL
			2.04	N

Reaction Buffer (used in incorporation reaction)

Ingredients	Amount	Unit	Conc.

Sterile Water for Injection (SWFI)	130.00	g	N/A
Sodium L-Ascorbate	7.80	g	57.25	mg/mL
Gentisic acid	2.60	g	19.08	mg/mL
2N NaOH	6.24	g	3.74	mg/mL

Formulation Buffer (used to prepare ¹⁷⁷Lu-DOTATATE

pharmaceutical composition from incorporation reaction mixture)

Ingredients	Amount	Unit	Conc.

Reaction buffer	120.00	g	N/A
DTPA	47.10	mg	0.38	mg/mL
2N NaOH	2.40	g	5.27	mg/mL
Sodium L-Ascorbate		g	56.13	mg/mL
Gentisic Acid		g	18.71	mg/mL

INCORPORATION BY REFERENCE

The entire contents of all patents, published patent applications and other disclosures cited herein are hereby expressly incorporated herein in their entireties by reference.

Claims

What is claimed is:

1. A pharmaceutical composition comprising:

(a) a complex of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE);

(b) one or more stabilizer compounds in a concentration of at least 25 mg/mL.

2. The composition of claim 1 wherein the complex of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) is present in an amount of at least 5 mCi/mL.

3. The composition of claim 1 wherein the complex of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) is present in an amount of at least 10 mCi/mL.

4. The composition of any one of claims 1 through 3 wherein the composition comprises two or more stabilizers.

5. The method of any one of claims 1 through 4 wherein the one or more stabilizers comprise one or more ascorbate compounds.

6. The method of any one of claims 1 through 5 wherein the one or more stabilizers comprise one or more gentisate compounds.

7. The composition of any one of claims 1 through 6 wherein the composition comprises at least one ascorbate compound and at least one gentisate compound.

8. The composition of any one of claims 1 through 7 wherein the one or more stabilizer compounds are present in an amount of 30 mg/mL or greater.

9. The composition of any one of claims 1 through 7 wherein the one or more stabilizer compounds are present in an amount of 40 mg/mL or greater.

10. The composition of any one of claims 1 through 7 wherein the one or more stabilizer compounds are present in an amount of 50 mg/mL or greater.

11. The composition of any one of claims 1 through 10 wherein the composition is an aqueous formulation.

12. The composition of any one of claims 1 through 11 wherein the complex of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) corresponds to the following structure:

13. The pharmaceutical composition of any one of claims 1 through 12 wherein the radiochemical purity of the composition is 95% or greater for 3 days by HPLC analysis.

14. The pharmaceutical composition of any one of claims 1 through 12 wherein the radiochemical purity of the composition is 95% or greater for 5 days by HPLC analysis.

15. A method for preparing ¹⁷⁷Lu-DOTATATE, comprising:

admixing lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) in the presence of one or more stabilizer compounds,

wherein the one or more stabilizer compounds are present in an amount of 10 mg/mL or greater,

thereby forming a complex of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE).

16. The method of claim 15 wherein an aqueous formulation comprising tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) is admixed with lutetium-177.

17. The method of claim 15 or 16 wherein the one or more stabilizer compounds are present in an amount of 20 mg/mL or greater.

18. The method of any one of claims 15 through 17 wherein the one or more stabilizer compounds are present in an amount of 30 mg/mL or greater.

19. The method of any one of claims 15 through 17 wherein the one or more stabilizer compounds are present in an amount of 40 mg/mL or greater.

20. The method of any one of claims 15 through 17 wherein the one or more stabilizer compounds are present in an amount of 50 mg/mL or greater.

21. The method of any one of claims 15 through 20 wherein each of the one or more stabilizer compounds are present in an amount of 10 mg/mL or greater.

22. The method of any one of claims 15 through 21 wherein the one or more stabilizers comprise one or more ascorbate compounds.

23. The method of any one of claims 15 through 22 wherein the one or more stabilizers comprise one or more gentisate compounds.

24. The method of any one of claims 15 through 23 wherein the one or more stabilizers comprise one or more gentisate compounds in an amount of at least 10 mg/mL and one or more ascorbate compounds in an amount of at least 30 mg/mL.

25. The method of any one of claims 15 through 23 wherein the one or more stabilizers comprise one or more gentisate compounds in an amount of at least 15 mg/mL and one or more ascorbate compounds in an amount of at least 40 mg/mL.

26. The method of any one of claims 15 through 25 wherein the admixture of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) is heated.

27. The method of claim 26 wherein the admixture is heated for 60 minutes or less.

28. The method of claim 26 wherein the admixture is heated for 50 minutes or less.

29. The method of claim 26 wherein the admixture is heated for 40 minutes or less.

30. The method of claim 26 wherein the admixture is heated for 30 minutes or less.

31. The method of claim 26 wherein the admixture is heated for 20 minutes or less.

32. The method of claim 26 wherein the admixture is heated for 15 minutes or less.

33. The method of any one of claims 15 through 32 wherein the admixture of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) is heated at about 80° C.±10° C.

34. The method of any one of claims 15 through 32 wherein the admixture lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) is heated at about 80° C.±5° C.

35. The method of any one of claims 15 through 34 further comprising cooling the admixture following heating.

36. The method of any one of claims 15 through 35 wherein following the admixing, the complex of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) is mixed with a formulation composition.

37. The method of claim 36 wherein the formulation composition is an aqueous composition that comprises one or more stabilizer compounds.

38. The method of claim 37 wherein the formulation composition comprises the one or more stabilizer compounds in an amount of 10 mg/mL or greater.

39. The method of claim 37 wherein the formulation composition comprises the one or more stabilizer compounds in an amount of 20 mg/mL or greater.

40. The method of claim 37 wherein the formulation composition comprises the one or more stabilizer compounds in an amount of 30 mg/mL or greater.

41. The method of claim 37 wherein the formulation composition comprises the one or more stabilizer compounds in an amount of 40 mg/mL or greater.

42. The method of claim 37 wherein the formulation composition comprises the one or more stabilizer compounds in an amount of 50 mg/mL or greater.

43. The method of any one of claims 36 through 42 wherein the formulation composition further comprises one or more sequestering agents.

44. The method of any one of claims 15 through 43 wherein incorporation of lutetium-177 with tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) provides ¹⁷⁷Lu-DOTATATE in greater than 98 mole percent.

45. The method of any one of claims 15 through 43 wherein incorporation of lutetium-177 with tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) provides ¹⁷⁷Lu-DOTATATE in greater than 99 mole percent.

46. The method of any one of claims 15 through 43 wherein an acidic aqueous formulation of lutetium-177 is admixed with the tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE).

47. The method of any one of claims 15 through 43 wherein a hydrogen halide or acid halide aqueous formulation of lutetium-177 is admixed with the tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE).

48. The method of any one of claims 1 through 47 wherein a hydrochloride acid aqueous formulation of lutetium-177 is admixed with the tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE).

49. A method for preparing ¹⁷⁷Lu-DOTATATE, comprising:

admixing lutetium-177 and an aqueous formulation comprising tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) in the presence of one or more ascorbate compounds,

wherein the one or more ascorbate compounds are present in an amount of 30 mg/mL or greater,

50. A method for preparing ¹⁷⁷Lu-DOTATATE, comprising:

admixing lutetium-177 and an aqueous formulation comprising tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) in the presence of one or more gentisate compounds,

wherein the one or more gentisate compounds are present in an amount of 10 mg/mL or greater,

51. A method for preparing ¹⁷⁷Lu-DOTATATE, comprising:

admixing lutetium-177 and an aqueous formulation comprising tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) in the presence of one or more stabilizer compounds that are selected from one or more ascorbate compounds and one or more gentisate compounds,

52. The method of claim 51 wherein the one or more stabilizer compounds are present in an amount of 20 mg/mL or greater.

53. The method of claim 51 wherein the one or more stabilizer compounds are present in an amount of 25 mg/mL or greater.

54. The method of claim 51 wherein the one or more stabilizer compounds are present in an amount of 30 mg/mL or greater.

55. The method of claim 51 wherein the one or more stabilizer compounds are present in an amount of 40 mg/mL or greater.

56. The method of claim 51 wherein the one or more stabilizer compounds are present in an amount of 50 mg/mL or greater.

57. The method of any one of claims 51 through 56 wherein the one more gentisate compounds are present in an amount of 10 mg/mL or greater.

58. The method of any one of claims 51 through 57 wherein the one or more stabilizers comprise one or more gentisate compounds in an amount of at least 10 mg/mL and one or more ascorbate compounds in an amount of at least 30 mg/mL.

59. The method of any one of claims 51 through 57 wherein the one or more stabilizers comprise one or more gentisate compounds in an amount of at least 15 mg/mL and one or more ascorbate compounds in an amount of at least 40 mg/mL.

60. The method of any one of claims 51 through 57 wherein the admixture of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) is heated.

61. The method of claim 60 wherein the admixture is heated for 40 minutes or less.

62. The method of claim 60 wherein the admixture is heated for 30 minutes or less.

63. The method of claim 60 wherein the admixture is heated for 20 minutes or less.

64. The method of any one of claims 51 through 63 wherein the admixture of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) is heated at about 80° C.±10° C.

65. The method of any one of claims 51 through 64 further comprising cooling the admixture following heating.

66. The method of any one of claims 51 through 65 wherein following the admixing, the complex of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) is mixed with a formulation composition.

67. The method of claim 66 wherein the formulation composition is an aqueous composition that comprises one or more stabilizer compounds.

68. The method of claim 67 wherein the formulation composition comprises the one or more stabilizer compounds in an amount of 10 mg/mL or greater.

69. The method of claim 67 wherein the formulation composition comprises the one or more stabilizer compounds in an amount of 20 mg/mL or greater.

70. The method of claim 67 wherein the formulation composition comprises the one or more stabilizer compounds in an amount of 30 mg/mL or greater.

71. The method of claim 67 wherein the formulation composition comprises the one or more stabilizer compounds in an amount of 40 mg/mL or greater.

72. The method of claim 67 wherein the formulation composition comprises the one or more stabilizer compounds in an amount of 50 mg/mL or greater.

73. The method of any one of claims 66 through 72 wherein the formulation composition further comprises one or more sequestering agents.

74. The method of any one of claims 51 through 73 wherein incorporation of lutetium-177 with tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) provides ¹⁷⁷Lu-DOTATATE in greater than 98 mole percent.

75. The method of any one of claims 51 through 73 wherein incorporation of lutetium-177 with tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) provides ¹⁷⁷Lu-DOTATATE in greater than 99 mole percent.

76. The method of any one of claims 51 through 75 wherein a hydrogen halide or acid halide aqueous formulation of lutetium-177 is admixed with the tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE).

77. The method of any one of claims 51 through 75 wherein a hydrochloride acid aqueous formulation of lutetium-177 is admixed with the tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE).

78. The method of any one of claims 51 through 77 wherein a composition is provided that comprises 1) a complex of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) and 2) one or more stabilizer compounds in a concentration of at least 25 mg/mL.

79. The method of claim 78 wherein the composition comprises the one or more stabilizer compounds in an amount of 30 mg/mL or greater.

80. The method of claim 78 wherein the composition comprises the one or more stabilizer compounds in an amount of 40 mg/mL or greater.

81. The method of claim 78 wherein the composition comprises the one or more stabilizer compounds in an amount of 50 mg/mL or greater.

82. The method of any one of claims 51 through 81 wherein composition comprises the complex of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) present in an amount of at least 5 mCi/mL.

83. The method of any one of claims 51 through 81 wherein composition comprises the complex of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) present in an amount of at least 10 mCi/mL.

84. The method of any one of claims 51 through 83 wherein the composition comprises two or more stabilizers.

85. The method of any one of claims 51 through 84 wherein the one or more stabilizers comprise one or more ascorbate compounds.

86. The method of any one of claims 51 through 85 wherein the one or more stabilizers comprise one or more gentisate compounds.

87. A method of any one of claims 15 through 86 further comprising administering a composition comprising a complex of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) to a patient in need thereof.

88. A method for treating a patient suffering from cancer, comprising:

(a) admixing lutetium-177 and an aqueous formulation comprising tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) in the presence of one or more stabilizer compounds that are selected from one or more ascorbate compounds and one or more gentisate compounds, thereby forming a complex of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE), and

wherein a composition is provided that comprises 1) a complex of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) and 2) one or more stabilizer compounds in a concentration of at least 25 mg/mL; and

b) administering the composition to the patient.

89. The method of claim 88 wherein the composition comprises the one or more stabilizer compounds in an amount of 30 mg/mL or greater.

90. The method of claim 88 wherein the composition comprises the one or more stabilizer compounds in an amount of 40 mg/mL or greater.

91. The method of claim 88 wherein the composition comprises the one or more stabilizer compounds in an amount of 50 mg/mL or greater.

92. The method of any one of claims 88 through 91 wherein composition comprises the complex of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) present in an amount of at least 5 mCi/mL.

93. The method of any one of claims 88 through 91 wherein composition comprises the complex of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) present in an amount of at least 10 mCi/mL.

94. The method of any one of claims 88 through 93 wherein the composition comprises two or more stabilizers.

95. The method of any one of claims 88 through 94 wherein the one or more stabilizers comprise one or more ascorbate compounds.

96. The method of any one of claims 88 through 95 wherein the one or more stabilizers comprise one or more gentisate compounds.

97. The method of any one of claims 15 through 96 further comprising determining purity of an ascorbate compound prior to admixing the ascorbate compound with lutetium-177 and/or tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) and/or a complex of lutetium-177 and/or tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE).

98. The method of claim 97 comprising determining the ascorbate compound is effective in an incorporation reaction and admixing the effective ascorbate compound with lutetium-177 and/or tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) and/or a complex of lutetium-177 and/or tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE).

99. The method of claim 97 or 98 wherein the ascorbate compound is an ascorbate salt.

100. The method of any one of claims claim 97 through 99 wherein the ascorbate compound is sodium ascorbate.

101. The method of any one of claims 15 through 100 wherein the complex of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) corresponds to the following structure:

102. The composition or method of any one of claims 1 through 101 wherein the complex of lutetium-177 and tetraazacyclododecane tetra-acetic acid-octreotate (DOTATATE) is ¹⁷⁷Lu-DOTATATE (¹⁷⁷Lu-octreotate).

103. ¹⁷⁷Lu-DOTATATE obtainable by a method of any one of claims 15-102.

104. ¹⁷⁷Lu-DOTATATE obtained by a method of any one of claims 15-102.

105. A pharmaceutical composition comprising ¹⁷⁷Lu-DOTATATE of claim 103 or 104.

106. The pharmaceutical composition of claim 105 wherein the radiochemical purity of the composition is 95% or greater for 3 days or more at 25° C.

107. The pharmaceutical composition of claim 105 wherein the radiochemical purity of the composition is 95% or greater for 5 days or more at 25° C.

108. The pharmaceutical composition of any one of claims 105 through 107 wherein the composition comprises one or more stabilizer compounds in an amount of 10 mg/mL or greater.

109. The pharmaceutical composition of any one of claims 105 through 107 wherein the composition comprises one or more stabilizer compounds in an amount of 20 mg/mL or greater.

110. The pharmaceutical composition of any one of claims 105 through 107 wherein the composition comprises one or more stabilizer compounds in an amount of 25 mg/mL or greater.

111. The pharmaceutical composition of any one of claims 105 through 107 wherein the composition comprises one or more stabilizer compounds in an amount of 30 mg/mL or greater.

112. The pharmaceutical composition of any one of claims 105 through 107 wherein the composition comprises one or more stabilizer compounds in an amount of 40 mg/mL or greater.

113. The pharmaceutical composition of any one of claims 105 through 107 wherein the composition comprises one or more stabilizer compounds in an amount of 50 mg/mL or greater.

114. The pharmaceutical composition of any one of claims 108 through 113 wherein the one or more stabilizers comprise one or more ascorbate compounds.

113. The pharmaceutical composition of any one of claims 108 through 114 wherein the one or more stabilizers comprise one or more gentisate compounds.

114. The pharmaceutical composition of any one of claims 108 through 113 wherein the one or more stabilizers comprise one or more gentisate compounds in an amount of at least 10 mg/mL and one or more ascorbate compounds in an amount of at least 30 mg/mL.

115. The pharmaceutical composition of any one of claims 108 through 113 wherein the one or more stabilizers comprise one or more gentisate compounds in an amount of at least 15 mg/mL and one or more ascorbate compounds in an amount of at least 40 mg/mL.

116. The pharmaceutical composition of any one of claims 108 through 113 wherein the composition comprises one or more ascorbate compounds are present in an amount of 30 mg/mL or greater.

117. The pharmaceutical composition of any one of claims 108 through 113 wherein the composition comprises one or more ascorbate compounds are present in an amount of 40 mg/mL or greater

118. The pharmaceutical composition of any one of claims 108 through 117 wherein the composition comprises one or more gentisate compounds are present in an amount of 10 mg/mL or greater.

119. A single dosage kit comprising ¹⁷⁷Lu-DOTATATE or pharmaceutical composition of any one of claims 1 through 14 or any one of claims 103 through 118.

120. A multiple dosage kit comprising ¹⁷⁷Lu-DOTATATE or pharmaceutical composition of any one of claims 1 through 14 or any one of claims 103 through 118.

121. A sealed container comprising ¹⁷⁷Lu-DOTATATE or pharmaceutical composition of any one of claims 1 through 14 or any one of claims 103 through 118.

122. A method of treating a subject suffering from cancer, comprising administering to the subject an effective amount of ¹⁷⁷Lu-DOTATATE or pharmaceutical composition of any one of claims 1 through 14 or any one of claims 103 through 121.

123. The method of claim 122 wherein the subject is suffering from neuroendocrine tumors.

124. The method of claim 122 wherein the subject is suffering from neuroendocrine tumors that originate from foregut, hindgut, midgut, lung, ovary, medulla, adrenal medulla, adrenal, kidney, pituitary, thyroid or paraganglia.

125. The method of claim 123 or 124 further comprising assessing the neuroendocrine tumors to be somatostatin receptor positive.

126. The method of 125 wherein the neuroendocrine tumors are demonstrated to be somatostatin receptor positive with ⁶⁸Ga-DOTATATE and a positron emission tomography scan.