Formulations and kits for radiation therapy and diagnostic imaging

文档序号：913523 发布日期：2021-02-26 浏览：12次中文

阅读说明：本技术 用于放射疗法和诊断成像的制剂和试剂盒 (Formulations and kits for radiation therapy and diagnostic imaging ) 是由 M·J·哈里斯肖恩·卢姆于 2019-04-11 设计创作，主要内容包括：公开了用稳定剂诸如L-甲硫氨酸和龙胆酸保护免受辐射分解的通式(II)的放射性药物化合物的水性制剂和试剂盒,其中所述化合物基于与放射性同位素诸如64-铜配位并且与四嗪基团连接的sarcophagine配体,所述四嗪基团用于与具有功能反应性基团诸如反式环辛烯的肿瘤靶向抗体反应,制备所述放射性配体制剂的工艺及其用于放射成像、诊断和治疗癌症的用途。(Aqueous formulations and kits of radiopharmaceutical compounds of general formula (II) protected from radiolysis with stabilizers such as L-methionine and gentisic acid are disclosed, wherein the compounds are based on coordination with a radioisotope such as 64-copper and with a tetrazinyl groupGroup-linked sarcophagine ligands, the tetrazine groups for reaction with tumor-targeting antibodies having functional reactive groups such as trans-cyclooctene, processes for preparing the radioligand formulations and their use for radioimaging, diagnosis and treatment of cancer.)

1. An aqueous formulation comprising a compound of formula (I) or a salt thereof,

the formulation further comprises sodium chloride and one or more stabilizers selected from the group consisting of: thiosulfate, ascorbate, ascorbic acid, gentisate, gentisic acid, methionine, maltose, inositol, benzyl alcohol, trehalose, povidone, niacinamide, cysteine and salts thereof.

2. An aqueous formulation comprising a compound of formula (I) or a salt thereof,

the formulation further comprises:

sodium chloride;

one or more stabilizers selected from the group consisting of: thiosulfate, ascorbate, ascorbic acid, gentisate, gentisic acid, methionine, maltose, inositol, benzyl alcohol, trehalose, povidone, niacinamide, cysteine and salts thereof;

ethanol; and is

Wherein the stabilizer is L-methionine or its salt.

3. An aqueous formulation comprising a compound of formula (I) or a salt thereof,

the formulation further comprises:

sodium chloride;

ethanol; and

an acetate salt;

wherein the stabilizer is L-methionine or its salt.

4. The aqueous formulation of any one of claims 1 to 3, wherein the formulation has a pH of between about 4 to about 8.

5. The aqueous formulation of any one of claims 1 to 4, wherein the compound of formula (I) is a hydrochloride salt.

6. The aqueous formulation of any one of claims 1 to 5, wherein the gentisate is sodium gentisate.

7. The aqueous formulation of any one of claims 1 to 6, wherein the compound of formula (I) is complexed with Cu ions.

8. The aqueous formulation of claim 7, wherein the Cu ion is a radioisotope.

9. The aqueous formulation of claim 8, wherein the Cu radioisotope is selected from the group consisting of⁶⁰Cu、⁶¹Cu、⁶⁴Cu and⁶⁷cu.

10. A process for preparing an aqueous formulation comprising a compound of formula (I) or a salt thereof complexed with Cu ions, the process comprising the steps of:

i) preparing a buffered solution of acetate salt, wherein the buffered solution further comprises ethanol and one or more stabilizing agents selected from the group consisting of: thiosulfate, ascorbate, ascorbic acid, gentisate, gentisic acid, methionine, maltose, inositol, benzyl alcohol, trehalose, povidone, niacinamide, cysteine and salts thereof;

ii) dissolving a compound of formula (I) or a salt thereof in the buffer solution obtained from step I);

iii) adding a solution of Cu ions to the solution obtained from step ii);

iv) filtering the solution obtained from step iii) onto a stationary phase; and

v) washing the stationary phase of step iv) with ethanol and brine into a solution of L-methionine;

to recover an aqueous formulation comprising a compound of formula (I) or a salt thereof complexed with Cu ions.

11. The process of claim 10, wherein theCu ions are selected from⁶⁰Cu、⁶¹Cu、⁶⁴Cu and⁶⁷a Cu radioisotope from the group consisting of Cu.

12. An aqueous formulation prepared by the process of claim 10 or 11.

13. A kit comprising a compound of formula (II) or a pharmaceutically acceptable salt thereof complexed with Cu ions:

the kit comprises:

a container comprising a compound of formula (II) or a pharmaceutically acceptable salt thereof,

a container containing a solution of Cu ions; and

instructions for preparing an aqueous formulation of a compound of formula (II) or a salt thereof complexed with Cu ions, the instructions comprising adding a solution of ethanol and one or more stabilizers selected from the group consisting of: thiosulfate, ascorbate, ascorbic acid, gentisate, gentisic acid, methionine, maltose, inositol, benzyl alcohol, trehalose, povidone, niacinamide, cysteine and salts thereof,

wherein:

R¹is H, optionally substituted alkyl or optionally substituted amino, and

R²、R³and R⁴Is a linker group; and is

R⁵Is H, optionally substituted alkyl, optionally substituted aryl or optionally substituted heteroaryl.

14. The kit of claim 13, further comprising the addition of L-methionine or a salt thereof.

15. The kit of claim 13 or 14, further comprising the addition of sodium chloride.

16. The kit of any one of claims 13 to 15, wherein the compound of formula (II) or salt thereof is lyophilized.

17. The kit according to any one of claims 13 to 16, wherein R¹Is CH₃Or NH₂。

18. The kit according to any one of claims 13 to 17, wherein R²Including benzylidene groups.

19. The kit according to any one of claims 13 to 18, wherein R²Including an aminobenzylidene group.

20. The kit according to any one of claims 13 to 19, wherein R³Including a propylene group.

21. The kit according to any one of claims 13 to 20, wherein R²Comprising a benzylidene group, and R³Including a propylene group.

22. The kit according to any one of claims 13 to 21, wherein R⁴Including phenylene groups.

23. The kit according to any one of claims 13 to 22, wherein the compound of formula (II) is:

24. the kit according to any one of claims 13 to 23, wherein the compound of formula (II) is:

25. the kit of any one of claims 13 to 24, wherein the Cu ion is a radioisotope.

26. The kit of claim 25, wherein the Cu ions are selected from the group consisting of⁶⁰Cu、⁶¹Cu、⁶⁴Cu and⁶⁷a radioisotope of the group consisting of Cu.

27. The kit of any one of claims 13 to 26, wherein the kit further comprises a container of a compound capable of binding to a tumor or cancer site and comprising a functional group that reacts with the tetrazine moiety of the compound of formula (II).

28. An aqueous formulation of a compound of formula (II) or a salt thereof complexed with Cu ions, prepared according to the kit of any one of claims 13 to 26.

29. A method for radioimaging, diagnosing or treating cancer, the method comprising administering the aqueous formulation of any one of claims 1 to 9, 12 or 28 to a subject in need thereof.

FIELD

The present invention relates to formulations and kits of compounds for use in radiotherapy and diagnostic imaging.

Background

The macrocyclic ligands can be used as radiopharmaceuticals in applications such as radiotherapy, or as diagnostic ligands for tumors or cancers. Particularly useful are radiolabeled ligands, which exhibit some propensity for selectively targeting specific sites in the body (e.g., specific receptors), may retain the radioisotope, and then deliver the radioisotope to the desired site of action. This requires that the ligand comprises a component that complexes the radioisotope and an additional component that targets the desired site. Traditionally, ligands are directly targeted to specific receptors specific to a particular site, such as a tumor or cancer site. Such direct targeting relies on the interaction between ligand and receptor being specific for a particular ligand and receptor, and the binding is sufficiently strong. In the event that the ligand is unable to bind strongly, the ligand (and accompanying radioisotope) may be recycled, causing unwanted damage to other sites by the radioisotope.

A known problem associated with macrocyclic ligands is that they can be complex organic compounds with multiple functional groups and subsequent reaction centers. This may mean that the ligand itself may decompose under specific conditions and become useless. Another problem associated with the ligand itself is that it may have limited solubility in a particular solvent. In addition to solubility problems, ligands may also be unstable and decompose in specific solvents. Ideally, the ligand is sufficiently soluble and stable in a pharmaceutically acceptable solvent in which it is administered for therapeutic purposes. Even if the ligand is sufficiently stable and soluble in a given solvent, the ligand complexed with the metal ion may have different properties and thus exhibit different stability and solubility profiles.

In the case of complexation of macrocyclic ligands with metal ions as radioisotopes, this may lead to radiolysis (radiolysis). This is caused by spontaneous radioactive decay and subsequent release of radiation from the radioisotope, such that the energy released may be sufficient to cause cleavage of the bond and result in destruction of the ligand. In addition to reducing the overall efficacy of the complexed ligand, the release of the radioisotope also results in the delivery of radiation to unwanted sites.

Since many radiopharmaceuticals are designed for parenteral administration, i.e. as injections (injectable) and usually as solutions, the ligand itself must be soluble in a pharmaceutically acceptable solvent or carrier. As is known in the art, the solubility of a particular compound in any given solvent may be unpredictable. While the solubility of a particular compound in a particular solvent may be known, the solubility of analogs of the compound in different solvent systems may vary considerably. This then presents additional difficulties for those seeking to develop formulations of the compounds, and in particular pharmaceutically acceptable injectable formulations.

Pharmaceutical formulations typically include one or more excipients that affect the compound in some way, such as enhancing the solubility of the compound or increasing the stability of the compound when in solution. Alternatively, additional excipients may be used to provide other features to the formulation, such as preservatives, buffers, and the like.

Although thousands of formulations of ligand-radioisotope complexes have been documented, there is no expectation that excipients used in such formulations will provide the solubility and bioavailability required for any newly developed complex. Furthermore, one cannot expect that a particular combination of excipients will further prevent or minimize dissociation of the radioisotope or minimize the occurrence of radiolysis.

Thus, there is a need to tailor the desired formulation of ligand-radioisotope complexes so as to exhibit the necessary stability associated with radiolysis and dissociation of the radioisotope while also being pharmaceutically acceptable. The present invention seeks to address these problems associated with particular ligand complexes.

The time for which the ligand-radioisotope complex is effective is ideally maximized. This would require preparation of a formulation comprising the ligand-radioisotope complex immediately prior to administration to a patient. While this is possible for patients located near synchrotrons or other radioisotope sources, for many other patients this will not be the case. These patients may need to transport the formation (format) over a significant distance, which reduces the time for the ligand-radioisotope complex to be effective. Thus, there is a need for a way to provide ligand-radioisotope complexes to facilities that are located at considerable distances from the source of the radioisotope.

Summary of The Invention

The present invention relates to formulations and kits of compounds that can be used as ligands, however the ligands are not necessarily targeted to the intended tumor or cancer site. The compounds of the formulations and kits disclosed herein do not bind directly to the tumor or cancer site, but rather target an intermediate compound that binds to the tumor or cancer site. The compounds of the kits and formulations comprise functional groups that can selectively react with the intermediate compounds. The advantage of this approach is that the compounds of the formulation and kit can then bind more selectively and more safely to the intermediate compound (as compared to binding directly to the tumor or cancer site), resulting in greater efficacy and less loss of the administered formulation due to diffusion or weak binding to the target site. This then means that the dose administered by the formulation can be optimised so that smaller doses are administered without any loss of overall efficacy. This also results in smaller amounts of compound and radioisotope being administered or lost.

According to a first aspect of the present invention there is provided an aqueous formulation comprising a compound of formula (I) or a salt thereof,

In an embodiment of the first aspect, the aqueous formulation comprises ethanol.

In another embodiment of the first aspect, the aqueous formulation comprises L-methionine or a salt thereof as a stabilizer.

According to a second aspect of the present invention there is provided an aqueous formulation comprising a compound of formula (I) or a salt thereof,

the formulation further comprises:

sodium chloride;

ethanol; and is

Wherein the stabilizer is L-methionine or its salt.

In an embodiment of the second aspect, the aqueous formulation comprises an acetate salt.

According to a third aspect of the present invention there is provided an aqueous formulation comprising a compound of formula (I) or a salt thereof,

the formulation further comprises:

sodium chloride;

ethanol; and

an acetate salt;

wherein the stabilizer is L-methionine or its salt.

According to a fourth aspect of the present invention there is provided a process for the preparation of an aqueous formulation comprising a compound of formula (I) or a salt thereof complexed with Cu ions, the process comprising the steps of:

i) preparing a buffered solution of acetate salt, wherein the buffered solution further comprises ethanol and one or more stabilizers selected from the group consisting of: thiosulfate, ascorbate, ascorbic acid, gentisate, gentisic acid, methionine, maltose, inositol, benzyl alcohol, trehalose, povidone, niacinamide, cysteine and salts thereof;

ii) dissolving the compound of formula (I) or a salt thereof in the buffer solution obtained from step I);

iii) adding a solution of Cu ions to the solution obtained from step ii);

iv) filtering the solution obtained from step iii) onto a stationary phase; and

v) washing the stationary phase of step iv) with ethanol and brine into a solution of L-methionine;

to recover an aqueous formulation comprising a compound of formula (I) or a salt thereof complexed with Cu ions.

According to a fifth aspect of the present invention there is provided an aqueous formulation prepared by the process as defined in the earlier aspect.

According to a sixth aspect of the present invention there is provided a kit comprising a compound of formula (II) or a pharmaceutically acceptable salt thereof complexed with Cu ions:

the kit comprises:

a container comprising a compound of formula (II) or a pharmaceutically acceptable salt thereof,

a container containing a solution of Cu ions; and

wherein:

R¹is H, optionally substituted alkyl or optionally substituted amino;

R²、R³and R⁴Is a linker group; and is

R⁵Is H, optionally substituted alkyl, optionally substituted aryl or optionally substituted heteroaryl.

In an embodiment of the sixth aspect, the kit further comprises a container of a compound that can bind to a tumor or cancer site and comprises a functional group that reacts with the tetrazine moiety of the compound of formula (II).

According to a seventh aspect of the present invention there is provided an aqueous formulation of a compound of formula (II) or a salt thereof complexed with Cu ions prepared according to the kit as defined in the seventh aspect.

According to an eighth aspect of the present invention there is provided a method for radioimaging, diagnosing or treating cancer, the method comprising administering to a subject in need thereof an aqueous formulation according to any one of the first to third, fifth or seventh aspects.

Detailed description of the invention

The present invention relates to stable formulations of macrocyclic ligands and their metal complexes. The present inventors have found that formulations of macrocyclic ligands as disclosed herein are stable relative to free ligands. The present inventors have also found that the formulation of metal complexes of macrocyclic ligands as disclosed herein minimizes dissociation of the complexed metal from the ligand and also minimizes radiolysis of the ligand in the case where the metal is a radioisotope.

Formulations of macrocyclic ligands and their metal complexes are stable in solution and under physiological conditions for extended periods of time. In the case of formulations comprising metal complexes of macrocyclic ligands, the stability of the formulation is related to the stability of the complex, where the metal is a radioisotope that can undergo dissociation, or where the complex can be decomposed under radiation. The stability of a formulation comprising a radioisotope-ligand complex may be measured by considering the radiochemical purity of the formulation. Radiochemical purity is defined as the amount of radioisotope complexed by the sarcophagine ligand expressed as a percentage of the total amount of radioisotope present in the formulation. The radioisotope may be present in the formulation as a complex with a sarcophagine ligand, as a free radioisotope or as part of a radiolysis product.

It has been found that tetrazine substituted ligands, such as ligands of formula (I), can target the trans-cyclooctene moiety and that tetrazine and trans-cyclooctene functional groups undergo an inverse electron demand Diels-Alder reaction (inverse electron demand Diels-Alder reaction) to produce corresponding click chemistry conjugates. Where the trans-cyclooctene functionality is installed in vivo on a target in a pretargeting strategy, this allows the tetrazine-substituted ligand complexed with the radioisotope to selectively bind to the trans-cyclooctene site and subsequently deliver the radioisotope to the desired site of action. Radioactivity derived from radioisotopes is commonly used in radiotherapy in the treatment of tumors. This pre-targeting strategy requires that antibodies that bind to the surface of the tumor be modified to include a trans-cyclooctene moiety, followed by administration of the modified antibodies to allow them to bind to the tumor site. A tetrazine-substituted ligand bearing a radioisotope is then administered, which selectively binds to the modified antibody, thereby delivering the radioisotope to the desired site of action.

Examples of tetrazine substituted ligands include compounds of formula (I):

the compounds of formula (I) are macrocyclic sarcophagine ligands in which the tetrazine moiety is bound to sarcophagine through a series of linking groups. sarcophagine or 3,6,10,13,16, 19-hexaazabicyclo [6.6.6] eicosane has an amino group at one terminal position wherein successive aminobenzyl, propyl and benzyl groups are linked by an amide bond. It will be understood by those skilled in the art that sarcophagine (or "sar") is a nitrogen-containing hexadentate macrocyclic ligand capable of complexing donor atoms, such as transition metal ions, and in the context of the present invention, Cu ions and radioisotopes thereof.

The compounds of formula (I) may be produced via a series of coupling reactions between diaminosarcophagine and a suitable coupling partner. Synthetic routes to compounds of formula (I) have been previously disclosed in mol. pharm.,2015,12,10, 3375.

The invention also contemplates the use of pharmaceutically acceptable salts of the compounds of formula (I) as part of the claimed formulations. Examples of the pharmaceutically acceptable salts of the compounds of formula (I) may include the corresponding acetate, sodium salt, hydrochloride, potassium salt, magnesium salt, calcium salt or ammonium salt. In embodiments, the compound of formula (I) is provided as an acetate salt.

The administrable formulation of the present invention comprises a complex of a compound of formula (I) or a salt thereof and a radioisotope. Radioisotopes may also be referred to as radionuclides, and may be metals or metal ions. The ligands of the present specification have been found to complex copper ions, particularly Cu²⁺Ionic aspects are particularly successful. Containing a radioisotope of copper ion of formula (I)Complexes have been previously disclosed in mol. pharm.,2015,12,10, 3375. It will also be appreciated by those skilled in the art that the complex of formula (I) and a radioisotope may be achieved by contacting a compound of formula (I) or a salt thereof with the radioisotope to be complexed, such that the compound of formula (I) or a salt thereof is complexed with the radioisotope. This may include mixing the compound of formula (I) or salt thereof and the radioisotope in a suitable solvent system, such as the solvent system specifically described herein.

In embodiments, the ligand is complexed with a Cu ion. The copper ions may be radioactive and thus a radionuclide or radioisotope of copper. In embodiments, the ligand is a ligand of the formula⁶⁰And (4) Cu complexing. In another embodiment, the ligand is a ligand of the formula⁶¹And (4) Cu complexing. In another embodiment, the ligand is a ligand of the formula⁶⁴And (4) Cu complexing. In another embodiment, the ligand is a ligand of the formula⁶⁷And (4) Cu complexing. In a preferred embodiment, the ligand is a ligand of the formula⁶⁴And (4) Cu complexing. In another preferred embodiment, the ligand is a ligand of the formula⁶⁷And (4) Cu complexing.

According to an aspect of the present invention there is provided an aqueous formulation comprising a compound of formula (I) or a salt thereof,

the formulation further comprises sodium chloride and a stabilizer or salt thereof.

In embodiments, sodium chloride is present in the formulation in an amount of about 0.6% (w/v) to 1.2% (w/v). In embodiments, sodium chloride is present in an amount of about 0.6% (w/v). In another embodiment, sodium chloride is present in an amount of about 0.7% (w/v). In another embodiment, sodium chloride is present in an amount of about 0.8% (w/v). In another embodiment, sodium chloride is present in an amount of about 0.9% (w/v). In another embodiment, sodium chloride is present in an amount of about 1.0% (w/v). In another embodiment, sodium chloride is present in an amount of about 1.1% (w/v). In another embodiment, sodium chloride is present in an amount of about 1.2% (w/v). In a preferred embodiment, sodium chloride is present in the formulation in an amount of about 0.9% (w/v). In other embodiments, the present invention also contemplates sodium chloride within a range between the amounts recited above.

The formulations of the present invention comprise one or more stabilizers as a component. The present inventors have determined that the presence of a stabilizer in the formulations of the invention helps to prevent or minimize radiolysis of the radiolabeled complex of formula (I). Examples of stabilizers which may be used in the formulations of the present invention include thiosulfates, ascorbates, ascorbic acid, gentisates, gentisic acid, methionine, maltose, inositol, benzyl alcohol, trehalose, povidone, niacinamide, cysteine and, where appropriate, salts thereof.

In embodiments, the formulations of the present invention comprise one or more stabilizers in a total amount of about 0.02% (w/v) to about 0.1% (w/v). In embodiments, the formulations of the present invention comprise one or more stabilizers in a total amount of about 0.02% (w/v). In another embodiment, the formulations of the present invention comprise one or more stabilizers in a total amount of about 0.025% (w/v). In another embodiment, the formulations of the present invention comprise one or more stabilizers in a total amount of about 0.03% (w/v). In another embodiment, the formulation of the present invention comprises one or more stabilizers in a total amount of about 0.035% (w/v). In another embodiment, the formulations of the present invention comprise one or more stabilizers in a total amount of about 0.04% (w/v). In another embodiment, the formulations of the present invention comprise one or more stabilizers in a total amount of about 0.045% (w/v). In another embodiment, the formulation of the present invention comprises one or more stabilizers in a total amount of about 0.05% (w/v). In another embodiment, the formulations of the present invention comprise one or more stabilizers in a total amount of about 0.055% (w/v). In another embodiment, the formulations of the present invention comprise one or more stabilizers in a total amount of about 0.6% (w/v). In another embodiment, the formulations of the present invention comprise one or more stabilizers in a total amount of about 0.065% (w/v). In another embodiment, the formulations of the present invention comprise one or more stabilizers in a total amount of about 0.07% (w/v). In another embodiment, the formulation of the present invention comprises one or more stabilizers in a total amount of about 0.075% (w/v). In another embodiment, the formulation of the present invention comprises one or more stabilizers in a total amount of about 0.08% (w/v). In another embodiment, the formulations of the present invention comprise one or more stabilizers in a total amount of about 0.085% (w/v). In another embodiment, the formulations of the present invention comprise one or more stabilizers in a total amount of about 0.09% (w/v). In another embodiment, the formulations of the present invention comprise one or more stabilizers in a total amount of about 0.095% (w/v). In another embodiment, the formulations of the present invention comprise one or more stabilizers in a total amount of about 0.1% (w/v). In other embodiments, the present invention also contemplates one or more stabilizers present in a range between the amounts recited above.

In an embodiment, a formulation comprising a compound of formula (I) comprises L-methionine or a salt thereof as a stabilizer. The term L-methionine as used herein refers to an amino acid with an S-methyl thioether side chain. The addition of L-methionine to the formulations of the present invention also enhances the stability of the formulation by preventing or minimizing radiolysis of the radiolabeled complex of formula (I), thereby increasing the radiochemical purity of the formulation. The inventors have found that the addition of L-methionine to a formulation comprising a compound of formula (I) and Cu ions, wherein the Cu ions are a radioisotope of Cu, allows to obtain a formulation with a higher initial radioactivity. This means that the dose of radioactivity delivered by a formulation comprising a compound of formula (I) and a complexed Cu radioisotope can be higher than the same formulation without the addition of L-methionine.

In embodiments, L-methionine or a salt thereof is present in the formulation in an amount from about 1mg/mL to about 4 mg/mL. In embodiments, L-methionine or a salt thereof is present in the formulation in an amount of about 1.0 mg/mL. In another embodiment, L-methionine or a salt thereof is present in the formulation in an amount of about 1.5 mg/mL. In another embodiment, L-methionine or a salt thereof is present in the formulation in an amount of about 2.0 mg/mL. In another embodiment, L-methionine or a salt thereof is present in the formulation in an amount of about 2.5 mg/mL. In another embodiment, L-methionine or a salt thereof is present in the formulation in an amount of about 3.0 mg/mL. In another embodiment, L-methionine or a salt thereof is present in the formulation in an amount of about 3.5 mg/mL. In another embodiment, L-methionine or a salt thereof is present in the formulation in an amount of about 4.0 mg/mL.

In another embodiment, gentisic acid or a salt thereof is present in the formulation as a stabilizer. Gentisic acid is also known as 2, 5-dihydroxybenzoic acid, 5-hydroxy salicylic acid or hydroquinone carboxylic acid. Salts of gentisic acid may include sodium salts and sodium salt hydrates. Where relevant, any reference to gentisic acid may include a reference to a salt thereof.

In another embodiment, ascorbic acid or a salt thereof is present in the formulation as a stabilizer. Ascorbic acid is also known as L-ascorbic acid or vitamin C. Salts of ascorbic acid include sodium ascorbate, calcium ascorbate, potassium ascorbate, and sodium ascorbyl phosphate. Derivatives of ascorbic acid are also contemplated. These include fatty acid esters of ascorbic acid, such as the palmitate ester of ascorbic acid, i.e. ascorbyl palmitate.

According to another aspect of the present invention there is provided an aqueous formulation comprising a compound of formula (I) or a salt thereof, the formulation further comprising:

sodium chloride;

ethanol; and is

Wherein the stabilizer is L-methionine or its salt.

In another embodiment of the invention, the aqueous formulation comprises an acetate salt. The acetate salt of the aqueous formulation may be an acetate salt, which acts as a buffer salt. The acetate salt may be ammonium acetate or sodium acetate. The acetate salt may be provided as part of a buffer solution used as part of the formulation.

Thus, in yet another aspect of the present invention, there is provided an aqueous formulation comprising a compound of formula (I) or a salt thereof, the formulation further comprising:

sodium chloride;

ethanol;

wherein the stabilizer is L-methionine or a salt thereof; and

an acetate salt.

The formulations of the present invention have a pH of about 4 to about 8. It will be appreciated by those skilled in the art that the pH of the formulation is an inherent property of the formulation due to the combination of the compound of formula (I) or complex thereof with the remaining excipients of the formulation. The present inventors have found that this pH range provides optimal radiolabelling efficiency, as well as stability of the radiolabelled complex in formulation and upon administration in vivo.

In embodiments, the pH of the formulation is from about 4 to about 8. In embodiments, the pH of the formulation is about 4. In another embodiment, the pH of the formulation is about 4.5. In another embodiment, the pH of the formulation is about 5.0. In embodiments, the pH of the formulation is about 5.5. In another embodiment, the pH of the formulation is about 5.6. In another embodiment, the pH of the formulation is about 5.7. In another embodiment, the pH of the formulation is about 5.8. In another embodiment, the pH of the formulation is about 5.9. In another embodiment, the pH of the formulation is about 6.0. In another embodiment, the pH of the formulation is about 6.1. In another embodiment, the pH of the formulation is about 6.2. In another embodiment, the pH of the formulation is about 6.3. In another embodiment, the pH of the formulation is about 6.4. In another embodiment, the pH of the formulation is about 6.5. In another embodiment, the pH of the formulation is about 7.0. In another embodiment, the pH of the formulation is about 7.5. In another embodiment, the pH of the formulation is about 8.0. In a preferred embodiment, the pH of the formulation is about 6.0.

In a further embodiment, an aqueous formulation of the invention comprises a compound of formula (I) or a salt thereof complexed to Cu ions, about 10% (v/v) ethanol, about 0.9% (w/v) sodium chloride, about 0.06% gentisic acid or a salt thereof, and about 2.5mg/mL L-methionine or a salt thereof, wherein the formulation has a pH of about 6.0. In another embodiment, an aqueous formulation of the invention comprises a compound of formula (I) or a salt thereof complexed with Cu ions, about 10% (v/v) ethanol, about 0.9% (w/v) sodium chloride, no more than 0.056% gentisic acid or a salt thereof, and about 2.5mg/mL L-methionine or a salt thereof, wherein the formulation has a pH of about 6.0. In a further embodiment, an aqueous formulation of the invention comprises a compound of formula (I) or a salt thereof complexed to Cu ions, about 10% (v/v) ethanol, about 0.9% (w/v) sodium chloride, 0.056% gentisic acid or a salt thereof and about 2.5mg/mL L-methionine or a salt thereof, wherein the formulation has a pH of about 6.0. The skilled person will appreciate that the amount of the formula (I) -Cu ion complex present in the aqueous formulation may be varied to suit different needs.

In a further embodiment, the aqueous formulation of the invention comprises⁶⁴A Cu ion complexed compound of formula (I) or a salt thereof, about 10% (v/v) ethanol, about 0.9% (w/v) sodium chloride, about 0.06% gentisic acid or a salt thereof, and about 2.5mg/mL L-methionine or a salt thereof, wherein the formulation has a pH of about 6.0. In embodiments, the aqueous formulation of the present invention comprises⁶⁴A Cu ion complexed compound of formula (I) or a salt thereof, about 10% (v/v) ethanol, no more than 0.9% (w/v) sodium chloride, no more than 0.056% gentisic acid or a salt thereof, and about 2.5mg/mL L-methionine or a salt thereof, wherein the formulation has a pH of about 6.0. In a further embodiment, the aqueous formulation of the invention comprises⁶⁴A Cu ion complexed compound of formula (I) or a salt thereof, about 10% (v/v) ethanol, about 0.9% (w/v) sodium chloride, about 0.056% gentisic acid or a salt thereof, and about 2.5mg/mL L-methionine or a salt thereof, wherein the formulation has a pH of about 6.0.

In a further embodiment, the aqueous formulation of the invention comprises⁶⁷Chemical combination of formula (I) with Cu ion complexationOr a salt thereof, about 10% (v/v) ethanol, about 0.9% (w/v) sodium chloride, about 0.06% gentisic acid or a salt thereof, and about 2.5mg/mL L-methionine or a salt thereof, wherein the formulation has a pH of about 6.0. In embodiments, the aqueous formulation of the present invention comprises⁶⁷A Cu ion complexed compound of formula (I) or a salt thereof, about 10% (v/v) ethanol, no more than 0.9% (w/v) sodium chloride, no more than 0.056% gentisic acid or a salt thereof, and about 2.5mg/mL L-methionine or a salt thereof, wherein the formulation has a pH of about 6.0. In a further embodiment, the aqueous formulation of the invention comprises⁶⁷A Cu ion complexed compound of formula (I) or a salt thereof, about 10% (v/v) ethanol, about 0.9% (w/v) sodium chloride, about 0.056% gentisic acid or a salt thereof, and about 2.5mg/mL L-methionine or a salt thereof, wherein the formulation has a pH of about 6.0. In a further embodiment, the aqueous formulation of the invention comprises⁶⁷A Cu ion complexed compound of formula (I) or a salt thereof, about 10% (v/v) ethanol, about 0.9% (w/v) sodium chloride, about 0.056% gentisic acid or a salt thereof, and about 2.5mg/mL L-methionine or a salt thereof, wherein the formulation has a pH of about 6.0.

In a further embodiment, the aqueous formulation of the invention comprises⁶⁴A Cu ion complexed compound of formula (I) as an acetate salt, about 10% (v/v) ethanol, about 0.9% sodium chloride (w/v), about 0.06% gentisic acid or a salt thereof and about 2.5mg/mL L-methionine or a salt thereof, wherein the formulation has a pH of about 6.0. In another embodiment, the aqueous formulation of the present invention comprises⁶⁴A Cu ion complexed compound of formula (I) as an acetate salt, about 10% (v/v) ethanol, about 0.9% (w/v) sodium chloride, about 0.056% gentisic acid or a salt thereof and about 2.5mg/mL L-methionine or a salt thereof, wherein the formulation has a pH of about 6.0. In another embodiment, the aqueous formulation of the present invention comprises⁶⁴A Cu ion complexed compound of formula (I) as an acetate salt, about 10% ethanol, about 0.9% (w/v) sodium chloride, no more than 0.056% gentisic acid or a salt thereof and about 2.5mg/mL L-methionine or a salt thereof, wherein the formulation has a pH of about 6.0.

In a further embodiment, the aqueous formulation of the invention comprises⁶⁷A Cu ion complexed compound of formula (I) as an acetate salt, about 10% (v/v) ethanol, about 0.9% (w/v) sodium chloride, about 0.06% gentisic acid or a salt thereof and about 2.5mg/mL L-methionine or a salt thereof, wherein the formulation has a pH of about 6.0. In another embodiment, the aqueous formulation of the present invention comprises⁶⁷A Cu ion complexed compound of formula (I) as an acetate salt, about 10% (v/v) ethanol, about 0.9% (w/v) sodium chloride, about 0.056% gentisic acid or a salt thereof and about 2.5mg/mL L-methionine or a salt thereof, wherein the formulation has a pH of about 6.0. In another embodiment, the aqueous formulation of the present invention comprises⁶⁷A Cu ion complexed compound of formula (I) as an acetate salt, about 10% ethanol, about 0.9% (w/v) sodium chloride, no more than 0.056% gentisic acid or a salt thereof and about 2.5mg/mL L-methionine or a salt thereof, wherein the formulation has a pH of about 6.0.

In another embodiment, the formulation of the present invention may further comprise ascorbic acid or a salt thereof as an additional stabilizer.

The present inventors have found that the formulations provided according to the present invention have increased radiochemical purity. The increased radiochemical purity of the formulation comprising the compound of formula (I) and the Cu radioisotope is due to the use of one or more stabilizers in the formulation.

According to the present invention, there is provided,⁶⁴the preparation of the complex of Cu and the compound of formula (I) may have a radiochemical purity of at least about 90% for a period of at least 45 hours. This means that present in the formulation⁶⁴At least about 90% of the Cu radioisotope is complexed with the compound of formula (I) or salt thereof at least 45 hours after preparation of the formulation. Present in the formulation⁶⁴In the case where the Cu radioisotope is not complexed with the compound of formula (I) or a salt thereof,⁶⁴the Cu radioisotope may be free⁶⁴Cu ions are present either as part of the radiolysis products.

In an embodiment, the invention comprises⁶⁷The radiochemical purity of a preparation of a complex of Cu and a compound of formula (I) or a salt thereof is about 90% in a time of about 11 hours after preparation of the preparation. In another embodimentIn the invention, the invention comprises⁶⁷The radiochemical purity of a formulation of a complex of Cu and a compound of formula (I) or a salt thereof is about 91% in a time of about 11 hours after preparation of the formulation. In another embodiment, the invention comprises⁶⁷The radiochemical purity of a preparation of a complex of Cu and a compound of formula (I) or a salt thereof is about 92% in a time of about 11 hours after preparation of the preparation. In another embodiment, the invention comprises⁶⁷The radiochemical purity of a preparation of a complex of Cu and a compound of formula (I) or a salt thereof is about 93% at a time of about 11 hours after preparation of the preparation. In another embodiment, the invention comprises⁶⁷The radiochemical purity of a preparation of a complex of Cu and a compound of formula (I) or a salt thereof is about 94% in a time of about 11 hours after preparation of the preparation. In another embodiment, the invention comprises⁶⁷The radiochemical purity of a preparation of a complex of Cu and a compound of formula (I) or a salt thereof is about 95% in a time of about 11 hours after preparation of the preparation. In another embodiment, the invention comprises⁶⁷The radiochemical purity of a formulation of a complex of Cu and a compound of formula (I) or a salt thereof is about 96% at a time of about 11 hours after preparation of the formulation. In another embodiment, the invention comprises⁶⁷The radiochemical purity of a preparation of a complex of Cu and a compound of formula (I) or a salt thereof is about 97% in a time of about 11 hours after preparation of the preparation. In another embodiment, the invention comprises⁶⁷The radiochemical purity of a preparation of a complex of Cu and a compound of formula (I) or a salt thereof is about 98% in a time of about 11 hours after preparation of the preparation. In another embodiment, the invention comprises⁶⁷The radiochemical purity of a formulation of a complex of Cu and a compound of formula (I) or a salt thereof is about 99% in a time of about 11 hours after preparation of the formulation.

In an embodiment, the invention comprises⁶⁷The radiochemical purity of a formulation of a complex of Cu and a compound of formula (I) or a salt thereof is about 99% shortly after preparation of the formulation. In another embodiment, the invention comprises⁶⁷Radiochemical purity of a preparation of a complex of Cu and a compound of formula (I) or a salt thereof, about after preparation of the preparationAfter 1h it was about 99%. In another embodiment, the invention comprises⁶⁷The radiochemical purity of a preparation of a complex of Cu and a compound of formula (I) or a salt thereof is about 99% after about 3h after preparation of the preparation. In another embodiment, the invention comprises⁶⁷The radiochemical purity of a preparation of a complex of Cu and a compound of formula (I) or a salt thereof is about 99% after about 6h after preparation of the preparation. In another embodiment, the invention comprises⁶⁷The radiochemical purity of a preparation of a complex of Cu and a compound of formula (I) or a salt thereof is about 99% after about 9h of preparation. In another embodiment, the invention comprises⁶⁷The radiochemical purity of a preparation of a complex of Cu and a compound of formula (I) or a salt thereof is about 99% after about 12h after preparation of the preparation. In another embodiment, the invention comprises⁶⁷The radiochemical purity of a preparation of a complex of Cu and a compound of formula (I) or a salt thereof is about 99% after about 15h after preparation of the preparation. In another embodiment, the invention comprises⁶⁷The radiochemical purity of a preparation of a complex of Cu and a compound of formula (I) or a salt thereof is about 99% after about 18h after preparation of the preparation. In another embodiment, the invention comprises⁶⁷The radiochemical purity of a preparation of a complex of Cu and a compound of formula (I) or a salt thereof is about 99% after about 21h after preparation of the preparation. In another embodiment, the invention comprises⁶⁷The radiochemical purity of a preparation of a complex of Cu and a compound of formula (I) or a salt thereof is about 99% after about 24h after preparation of the preparation.

The invention also relates to kits of macrocyclic ligands. The kit comprises a container comprising a compound of formula (II) or a salt thereof, and a container comprising a solution of Cu ions. The present inventors have found that the kits disclosed herein allow for the preparation of formulations of ligand-radioisotope complexes, radioisotopes such as Cu radioisotopes, wherein dissociation of the radioisotope from the ligand and radiolysis due to the radioisotope is minimized. The kit, in turn, provides for the formulation of a ligand-radioisotope complex wherein the effective time for imaging or therapy using the complex is maximized. The kits of the invention also allow for the preparation of the formulation immediately prior to administration to a patient. Such in situ preparation increases the time that the formulation can be effective in vivo, which in turn provides improved efficacy of the radiolabeled complex for both therapeutic and diagnostic purposes.

According to another aspect of the present invention, there is provided a kit comprising a compound of formula (II) or a pharmaceutically acceptable salt thereof complexed with Cu ions:

the kit comprises:

a container comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof,

a container containing a solution of Cu ions; and

instructions for preparing an aqueous formulation of a compound of formula (I) or a salt thereof complexed with Cu ions, the instructions comprising adding a solution of ethanol and one or more stabilizers,

wherein:

R¹is H, optionally substituted alkyl or optionally substituted amino;

R²、R³and R⁴Is a linker group; and is

R⁵Is H, optionally substituted alkyl, optionally substituted aryl or optionally substituted heteroaryl.

Like the compound of formula (I), the compound of formula (II) is also a macrocyclic sarcophagine ligand (3,6,10,13,16, 19-hexaazabicyclo [6.6.6] eicosane, "sar") and contains a 1,2,4, 5-tetrazine moiety at a terminal position of the compound. The tetrazine moiety in both compounds of formula (I) and compounds of formula (II) consists of a six-membered ring in which the four atoms at positions 1,2,4 and 5 of the ring are nitrogen. This particular arrangement of nitrogen atoms around the six-membered ring results in an "s-tetrazine" which can participate in a click-type reaction with olefins. In particular, the tetrazine functionality of the compound of formula (I) or the compound of formula (II) may undergo a Diels-Alder cycloaddition with the trans-cyclooctene functionality with the tetrazine acting as a dienophile and the trans-cyclooctene acting as a diene. The anti-electron-requiring Diels-Alder cycloaddition between the tetrazine of formula (I) and the trans-cyclooctene occurs rapidly to provide a cyclized product. The rate at which the cycloaddition reaction between tetrazine and trans-cyclooctene proceeds may be affected by any substituent on each of these groups. The compounds of formula (II) may have additional substituents in the para position of the tetrazine ring. Tetrazines can also undergo click-type reactions with other unsaturated compounds, which can lead to the production of undesirable by-products. Given the many heteroatoms present in tetrazines, the functional groups and subsequently the compounds themselves may undergo decomposition.

As used herein, the term "alkyl" refers to a group or part of a group containing a straight or branched aliphatic hydrocarbon group, preferably C₁-C₁₂Alkyl, more preferably C₁-C₁₀Alkyl, most preferably C₁-C₆Unless otherwise indicated. Suitable straight-chain and branched C₁-C₆Examples of alkyl substituents include methyl, ethyl, n-propyl, 2-propyl, n-butyl, sec-butyl, tert-butyl, hexyl and the like. The group may be a terminal group or a bridging group.

As used herein, the term "alkenyl" refers to a group or part of a group that is an aliphatic hydrocarbon group containing at least one carbon-carbon double bond, and that may be straight or branched, preferably having 2-12 carbon atoms, more preferably 2-10 carbon atoms, most preferably 2-6 carbon atoms in the normal chain (normal chain). The group may contain more than one double bond in the normal chain and is independently E or Z with respect to the orientation of each. Exemplary alkenyl groups include, but are not limited to, ethenyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, and nonenyl. The group may be a terminal group or a bridging group.

As used herein, the term "alkynyl" refers to a group or part of a group that is an aliphatic hydrocarbon group containing a carbon-carbon triple bond, and that may be straight or branched, preferably having from 2 to 12 carbon atoms, more preferably 2 to 10 carbon atoms, more preferably 2 to 6 carbon atoms in the normal chain. Exemplary structures include, but are not limited to, ethynyl and propynyl. The group may be a terminal group or a bridging group.

As used herein, the term "aryl" refers to a group or part of a group that is (i) an optionally substituted monocyclic aromatic carbocyclic ring or fused polycyclic aromatic carbocyclic ring (ring structure having ring atoms that are all carbon), preferably from 5 to 12 atoms per ring, wherein examples of aryl groups include phenyl, naphthyl, and the like; (ii) an optionally substituted partially saturated bicyclic aromatic carbocyclic moiety wherein the phenyl group and C₅-C₇Cycloalkyl radicals or C₅-C₇Cycloalkenyl groups are fused together to form a cyclic structure, such as tetrahydronaphthyl, indenyl, or indanyl. The group may be a terminal group or a bridging group. Typically, the aryl group is C₆-C₁₈An aryl group.

As used herein, the term "cycloalkyl" refers to a saturated monocyclic carbocyclic ring or a fused carbocyclic or spiro polycyclic carbocyclic ring, preferably containing from 3 to 9 carbons per ring, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like, unless otherwise specified. The term "cycloalkyl" includes monocyclic ring systems such as cyclopropyl and cyclohexyl, bicyclic ring systems such as decahydronaphthalene, and polycyclic ring systems such as adamantane. Cycloalkyl radicals are usually C₃-C₉A cycloalkyl group. The group may be a terminal group or a bridging group.

As used herein, the term "halogen" represents chlorine, fluorine, bromine or iodine.

As used herein, the term "heteroalkyl" refers to a straight or branched chain alkyl group, preferably having from 2 to 12 carbons in the chain, more preferably 2 to 6 carbons, in which one or more carbon atoms (and any associated hydrogen atoms) are each independently selected from S, O, P andNR ', wherein R' is selected from the group consisting of: H. optionally substituted C₁-C₁₂Alkyl, optionally substituted C₃-C₁₂Cycloalkyl, optionally substituted C₆-C₁₈Aryl and optionally substituted C₁-C₁₈A heteroaryl group. Exemplary heteroalkyl groups include alkyl ethers, secondary and tertiary alkyl amines, amides, alkyl sulfides, and the like. Examples of heteroalkyl radicals also include hydroxy C₁-C₆Alkyl radical, C₁-C₆Alkoxy radical C₁-C₆Alkyl, amino C₁-C₆Alkyl radical, C₁-C₆Alkylamino radical C₁-C₆Alkyl and di (C)₁-C₆Alkyl) amino C₁-C₆An alkyl group. The group may be a terminal group or a bridging group.

As used herein, the term "heteroaryl" refers to a group comprising an aromatic ring (preferably a 5-or 6-membered aromatic ring) having one or more heteroatoms as ring atoms in the aromatic ring, wherein the remaining ring atoms are carbon atoms. Suitable heteroatoms include nitrogen, oxygen, and sulfur. Examples of heteroaryl groups include thiophene, benzothiophene, benzofuran, benzimidazole, benzoxazole, benzothiazole, benzisothiazole, naphtho [2,3-b]Thiophene, furan, isoindoline, xanthone (xanthene), phenoxazine (phenoxatine), pyrrole, imidazole, pyrazole, pyridine, pyrazine, pyrimidine, pyridazine, tetrazole, indole, isoindole, 1H-indazole, purine, quinoline, isoquinoline, phthalazine, naphthyridine, quinoxaline, cinnoline, carbazole, phenanthridine, acridine, phenazine, thiazole, isothiazole, phenothiazine, oxazole, isoxazole, furazan, phenoxazine, 2-pyridyl, 3-pyridyl or 4-pyridyl, 2-quinolyl, 3-quinolyl, 4-quinolyl, 5-quinolyl or 8-quinolyl, 1-isoquinolyl, 3-isoquinolyl, 4-isoquinolyl or 5-isoquinolyl, 1-indolyl, 2-indolyl or 3-indolyl, and 2-thienyl or 3-thienyl. The heteroaryl group is typically C₅-C₁₈A heteroaryl group. The group may be a terminal group or a bridging group.

The term "optionally substituted" as used throughout the specification means that the group may or may not be further substituted or fused (so as to form a fused polycyclic system) with one or more non-hydrogen substituent groups. In certain embodiments, a substituent group is one or more groups independently selected from the group consisting of: halogen, ═ O, ═ S, -CN, -NO₂、-CF₃、-OCF₃Alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl, cycloalkylalkyl, heterocycloalkylalkyl, heteroarylalkyl, arylalkyl, cycloalkylalkenyl, heterocycloalkylalkenyl, arylalkenyl, heteroarylalkenyl, cycloalkylheteroalkyl, heterocycloalkylheteroalkyl, arylheteroalkyl, heteroarylheteroalkyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, alkoxycycloalkyl, alkoxyheterocycloalkyl, alkoxyaryl, alkoxyheteroaryl, alkoxycarbonyl, alkylaminocarbonyl, alkenyloxy, alkynyloxy, cycloalkoxy, cycloalkenyloxy, heterocycloalkoxy, heterocycloalkenyloxy, aryloxy, phenoxy, benzyloxy, heteroaryloxy, arylalkoxy, amino, alkylamino, amido, aminoalkyl, amino, and the like, Arylamino, sulfonylamino, sulfinylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, sulfinyl, alkylsulfinyl, arylsulfinyl, aminosulfinylaminoalkyl, -C (═ O) OH, -C (═ O) Ra, -C (═ O) ORa, -C (═ O) NR^aR^b、-C(＝NOH)R^a、-C(＝NR^a)NR^bR^c、-NR^aR^b、-NR^aC(＝O)R^b、-NR^aC(＝O)OR^b、-NR^aC(＝O)NR^bR^c、-NR^aC(＝NR^b)NR^cR^d、-NR^aSO₂R^b、-SR^a、-SO₂NR^aR^b、-OR^a、-OC(＝O)NR^aR^b、-OC(＝O)R^aAnd acyl, wherein R^a、R^b、R^cAnd R^dEach is independentThe locus is selected from the group consisting of: H. c₁-C₁₂Alkyl radical, C₁-C₁₂Haloalkyl, C₂-C₁₂Alkenyl radical, C₂-C₁₂Alkynyl, C₂-C₁₀Heteroalkyl group, C₃-C₁₂Cycloalkyl radical, C₃-C₁₂Cycloalkenyl radical, C₂-C₁₂Heterocycloalkyl radical, C₂-C₁₂Heterocycloalkenyl, C₆-C₁₈Aryl radical, C₁-C₁₈Heteroaryl and acyl, or R^a、R^b、R^cAnd R^dAny two or more of which, when taken together with the atoms to which they are attached, form a heterocyclic ring system having from 3 to 12 ring atoms.

In some embodiments, each optional substituent is independently selected from the group consisting of: halogen, ═ O, ═ S, -CN, -NO₂、-CF₃、-OCF₃Alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, alkoxyaryl, alkoxyheteroaryl, alkenyloxy, alkynyloxy, cycloalkoxy, cycloalkenyloxy, heterocycloalkoxy, heterocycloalkenyloxy, aryloxy, heteroaryloxy, arylalkyl, heteroarylalkyl, arylalkoxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, aminoalkyl, -COOH, -SH and acyl.

Examples of particularly suitable optional substituents include F, Cl, Br, I, CH₃、CH₂CH₃、OH、OCH₃、CF₃、OCF₃、NO₂、NH₂And CN.

As used herein, the term "C₁-C₁₂Alkenyl "refers to a divalent straight or branched aliphatic hydrocarbon group wherein the group has 1 to 12 carbon atoms in the chain.

As used herein, the term "arylene" refers to a divalent aromatic hydrocarbon group.

The term "pharmaceutically acceptable salts" refers to salts that retain the desired biological activity of the compounds identified above and includes pharmaceutically acceptable acid addition salts and base addition salts. Suitable pharmaceutically acceptable acid addition salts of the compounds of formula (I) or of the compounds of formula (II) may be prepared from inorganic acids or from organic acids. Examples of such inorganic acids are hydrochloric acid, sulfuric acid and phosphoric acid. Suitable organic acids may be selected from the organic acids of the aliphatic, cycloaliphatic, aromatic, heterocyclic carboxylic and sulfonic classes, examples of which are formic, acetic, propionic, succinic, glycolic, gluconic, lactic, malic, tartaric, citric, fumaric, maleic, alkyl sulfonic and aryl sulfonic acids. Additional information on pharmaceutically acceptable salts can be found in Remington's Pharmaceutical Sciences, 19 th edition, Mack Publishing co., Easton, PA 1995. In the case of solid agents, those skilled in the art will appreciate that the compounds, agents and salts of the present invention may exist in different crystalline or polymorphic forms, all of which are intended to be within the scope of the present invention and the indicated formula.

The compound of formula (II) may comprise a substituent at a terminal position of the cage, i.e. R in the compound of formula (II)¹. In some embodiments, R¹May be optionally substituted C₁-C₁₂An alkyl group. In other embodiments, R¹May be an optionally substituted amino group.

The compound of formula (II) comprises R²、R³And R⁴A plurality of linker groups as indicated.

In some embodiments, the linker R in the compound of formula (II)²Is absent. In some embodiments, R²Including arylene groups. In some embodiments, R²Also included are amino groups. In some embodiments, R²Including arylene groups and amino groups separated by alkylene groups. In some embodiments, R²Comprising one or more alkylene groups connecting the amino group and the arylene group to formula(I) The remainder of the compound of (1). In some embodiments, R²Is an aminobenzylidene group.

In some embodiments, the linker R in the compound of formula (II)³Is optionally substituted C₁-C₁₂An alkylene group. In embodiments, R³Is optionally substituted C₃An alkylene group. In embodiments, R³Is unsubstituted C₃An alkylene group.

In some embodiments, the linker R in the compound of formula (II)⁴Is an optionally substituted arylene group. In some embodiments, R⁴Is an optionally substituted benzylidene group.

The compound of formula (II) may also contain a substituent group at the 4-or para-position of the tetrazine ring, i.e. R in the compound of formula (II)⁵. In some embodiments, R⁵Is H. In some embodiments, R⁵Is an optionally substituted alkyl group. In some embodiments, R⁵Is an optionally substituted aryl group. In some embodiments, R⁵Is an optionally substituted heteroaryl group. In some embodiments, R⁵Is a haloalkyl group.

In embodiments, the compound of formula (II) is:

it is a methyl-substituted sarcophagine having a series of linkages between the sarcophagine and the tetrazine. This compound is also known as N1- (4- (1,2,4, 5-tetrazin-3-yl) benzyl) -N5- (8-methyl-3, 6,10,13,16, 19-hexaazabicyclo [6.6.6] eicosan-1-yl) glutaramide and is abbreviated MeCOSar-Tz.

The compound of formula (II) in the kit of the invention may be provided as a solid. In some embodiments, the compound of formula (I) may be provided in a container as a lyophilized solid. In some embodiments, the kits of the invention may comprise the compound of formula (II) in a container as a lyophilized solid. In some embodiments, the kits of the invention may comprise the compound of formula (II) as a lyophilized solid in a container, wherein additional additives are contained in the same container.

The kit of the present invention includes a container of copper (Cu) ions. In an embodiment, the kit of the invention comprises a container providing a solution of Cu ions. In some embodiments, the Cu ions are provided as a Cu salt. In some embodiments, the Cu salt is Cu^IAnd (3) salt. In other embodiments, the Cu salt is Cu^IIAnd (3) salt. In further embodiments, the Cu salt is Cu^IIIAnd (3) salt. In embodiments, the kits of the invention comprise a container that provides a solution of a Cu radionuclide. In embodiments, the kits of the invention comprise a container having a solution of a Cu radioisotope. In embodiments, the Cu radioisotope may be⁶⁰And (3) Cu. In another embodiment, the Cu radioisotope may be⁶¹And (3) Cu. In another embodiment, the Cu radioisotope may be⁶⁴And (3) Cu. In another embodiment, the Cu radioisotope may be⁶⁴And (3) Cu. In some embodiments, the Cu radioisotope is provided in a solution of hydrochloric acid. The concentration of the hydrochloric acid solution in which the Cu radioisotope is provided may be from 0.01M to 0.1M. In some embodiments, wherein the hydrochloric acid solution providing the Cu radioisotope is at a concentration of about 0.05M.

The kit of the invention further comprises instructions for preparing an aqueous formulation of the compound of formula (II) complexed with Cu ions from a container comprising the compound of formula (II) and a container comprising a solution of Cu ions. The Cu ions in the kit of the present invention may be provided as a solution. In some embodiments, the Cu ions are provided in the vessel as a solution of the corresponding chloride salt of the Cu ions. In some embodiments, the Cu ions are provided in the vessel as a hydrochloric acid solution of the corresponding chloride salt of the Cu ions. In some embodiments, the Cu ions are radioisotopes and are provided in a container that isolates radioactivity from the environment. In some embodiments, the Cu ions are radioisotopes provided in a container obtained from a synchrotron.

The kit of the present invention may further comprise a container consisting of a solution of ethanol and a stabilizer. Alternatively, the kit may provide instructions for adding a solution of ethanol and stabilizer. The present inventors have found that the presence of one or more stabilizers helps to prevent or minimize radiolysis of the complex of formula (II) and Cu in the aqueous formulation obtained from the kit of the present invention. With regard to the formulation of the compound of formula (I), examples of stabilizers that may be used in the kit of the present invention include thiosulfate, ascorbate, ascorbic acid, gentisate, gentisic acid, L-methionine, maltose, inositol, benzyl alcohol, trehalose, povidone, nicotinamide, cysteine and, where appropriate, salts thereof. In embodiments, the kit comprises a container comprising a solution of ethanol and a stabilizer. In another embodiment, a kit includes a container containing a solution of ethanol and one or more stabilizers. In embodiments, the stabilizing agent is gentisic acid or a salt thereof. In another embodiment, the stabilizer is L-methionine or a salt thereof.

The kit of the present invention may further comprise additional additives. Examples of additives include, but are not limited to, salts such as sodium chloride, buffers such as ammonium acetate, and similar compounds. In some embodiments, the kits of the invention comprise one or more additives, which are provided in separate containers. In some embodiments, the kits of the invention include one or more additives provided in containers intended for other components of the kit.

According to certain aspects of the present invention, the formulations and kits defined herein comprise ethanol. The ethanol used in the formulation may be anhydrous ethanol. Alternatively, the ethanol used in the formulation may not have undergone a drying process and may be hydrated. The ethanol is preferably pharmaceutical grade ethanol. The ethanol present in the formulations of the present invention may also help prevent radiolysis of the radiolabeled complex of formula (I) when combined with one or more stabilizers in the formulation.

In embodiments, ethanol is present in the formulation in an amount from about 7% (v/v) to about 13% (v/v). In embodiments, ethanol is present in the formulation in an amount of about 7% (v/v). In another embodiment, ethanol is present in the formulation in an amount of about 8% (v/v). In another embodiment, ethanol is present in the formulation in an amount of about 9% (v/v). In another embodiment, ethanol is present in the formulation in an amount of about 10% (v/v). In another embodiment, ethanol is present in the formulation in an amount of about 11% (v/v). In another embodiment, ethanol is present in the formulation in an amount of about 12% (v/v). In another embodiment, ethanol is present in the formulation in an amount of about 13% (v/v). In a preferred embodiment, ethanol is present in the formulation in an amount of about 10% (v/v). In other embodiments, the invention also contemplates ethanol within a range between the amounts recited above.

The kit of the invention further comprises ethanol. In some embodiments, the ethanol may be provided in a container of the kit. In some embodiments, the ethanol may be provided in a container with the stabilizer of the kit. In some embodiments, the ethanol may be provided in a container with one or more additional additives of the kit. In some embodiments, the ethanol may be provided in a container with the stabilizer and one or more additives of the kit. In some embodiments, the ethanol is not provided by the kit, but rather by the end user of the kit.

The kit of the invention may comprise further containers with further additives. Examples of such additives include organic and inorganic salts or solutions thereof. The container containing the further additive of the kit may provide the further additive as part of a solid or a solution. In embodiments, the kits of the invention further comprise sodium chloride or a solution thereof. In embodiments, the kits of the invention further comprise ammonium acetate or a solution thereof.

In embodiments, the kits of the invention comprise a container comprising a compound of formula (II) or a salt thereof, a container comprising a solution of Cu ions, and instructions for preparing an aqueous formulation of the compound of formula (II) or a salt thereof complexed with Cu ions. In embodiments, the kits of the invention comprise a container comprising a compound of formula (II) or a salt thereof, a container comprising a solution of Cu ions, a container comprising a solution of ethanol and one or more stabilizers, and instructions for preparing an aqueous formulation of the compound of formula (II) or a salt thereof complexed with Cu ions.