Methods and kits for preparing radionuclide complexes

Abstract

To provide a method for preparing a radioactive gallium complex for use in therapy or diagnosis.SOLUTION: There is provided a method for the preparation of a complex comprising a radioisotope of gallium for use in radiotherapy or medical imaging, wherein a gallium radioisotope solution obtained directly from a gallium radionuclide generator is added to a composition comprising a pharmaceutically acceptable buffer and a basic reagent to adjust the pH to between 3 and 8, wherein the composition further comprises a chelating agent capable of chelating radioactive gallium within said pH range and at moderate temperatures, wherein said chelating agent is optionally associated with a biological targeting agent.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to radioactive gallium ion beams for use in therapy or diagnosis, for example in molecular imaging. The present invention relates to methods for preparing ammonium complexes, kits for use in these methods, and materials for use therein. The present invention relates to novel compositions and molecular imaging and methods performed using the compositions or kits. and treatment. [Background technology]

[0002] Molecular imaging is a well-known and useful technique for in vivo diagnostics. is a molecular mechanism for determining gene expression, blood flow, physiological changes (e.g., pH), immune responses, and cellular transport. It can be used in a wide variety of ways, such as for 3D mapping of processes, detecting disease, to identify and diagnose, to select optimal treatment, and to monitor the effectiveness of treatment This can be used to get an early readout of the effect.

[0003] A number of distinct techniques are available, including positron emission tomography (PET), single photon emission tomography (SPE), (SPET), Optical (O1) Magnetic Resonance Imaging (MRI), X-ray Computed Tomography (C molecular luminescence imaging (MLI), such as T and Cerenkov luminescence imaging (CLI) In principle, a combination of these modalities can be used for imaging. PET / CT and SPET / CT ("multimodal imaging"), for example, are are emerging to offer improved clinical applications.

[0004] The radionuclides imaged by PET and SPET offer extremely high sensitivity and in vivo Contrast agents administered in small amounts (e.g., picomolar in vivo) that do not perturb molecular processes. Furthermore, the targeting principle of radionuclide imaging also has the advantage of It can also be applied to targeted delivery of radionuclide therapy. Typically, it is used in molecular imaging. or isotopes used as radionuclides in therapy must be pharmaceutically acceptable to the subject. The compound is incorporated into a molecule to produce a radioactive tracer.

[0005] Most radiotracers have relatively short half-lives and therefore require in situ administration under sterile conditions. must be produced, for example, in the radiopharmacy department of the hospital concerned. Hospitals have difficulty with this if they do not have specialized radiochemistry laboratories, and therefore A hospital's ability to offer treatments such as PET may be limited.

[0006] It can be difficult to prepare radiotracers with reactive functional groups. For example, Incorporation of radioisotopes into radiotracers disrupts protein structure and is undesirable. Reactive functional groups can involve high temperatures which would add unnecessary complexity to the labeling process. It may be desirable to include the radioactive tracer in It is necessary to provide a radioactive tracer that can be prepared by the labeling process at the time of use. requires the minimum number of manipulations of radioactive materials, the minimum need for expensive equipment to perform the manipulations, It is desirable that the preparation be as simple as possible and in the shortest possible time. Imaging conjugates (c) with improved functionality and enhanced molecular imaging properties onjugate) was created.

[0007] 1,4,7,10-Tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA) is a gallium-ion radioisotope used in molecular imaging and targeted radionuclide therapy. 68 (and others such as Ga-67, In-11, Cu-64, Lu-177, Y-90, etc. However, DOTA is a common chelating agent for radioisotopes of metals. 68 It has a long radiolabeling time of about 30 to 60 minutes (compared to the half-life of Ga, which is about 68 minutes), and Furthermore, chelation of gallium by DOTA derivatives In many cases, this can be damaging to any biological targeting agent associated with the biotracer. , requiring a high labeling temperature of about 95° C. and an acidic pH.

[0008] WO 2012 / 063028 pamphlet describes dissociation in a biological environment. A wide range of bifunctional compounds that can rapidly chelate radionuclides at room temperature while retaining stability. In addition to the metal chelating moiety, the bifunctional molecule is a targeting group that can target, for example, cells, tissues, or biomolecules throughout the body. They have reactive moieties for linking to functional groups such as . Kits containing these bifunctional molecules and radionuclides are also described.

[0009] However, residual problems arise with respect to the radionuclides themselves. These are generally: It is obtained by elution from a generator. 68 Many of these isotopes, such as Ga, However, they will only elute at low pH, e.g., less than 2, such as about 1. However, they were prepared either before or after the addition of a chelating agent to generate the radiotracer. In particular, gallium requires complex pretreatment procedures to raise the pH to a neutral to high pH range. It tends to precipitate out of solution in H and therefore requires special handling. ,for example 68 The eluate from the Ga generator is then added to the chelating agent before it can be contacted with the chelating agent. The sample is first subjected to a purification step by passage through a cation exchange cartridge. The recognition procedure is also complex, for example, adding buffers and acids along with the chelating compounds. and then subjecting the mixture to a relatively high temperature, e.g., 100°C, to achieve labeling. Heating may be necessary. The product is then dissolved in phosphate buffered saline (PBS) solution. The solution was diluted and passed through a sterile filter before being placed on a SEP-Pak C-18 cartridge. This may require further steps of passing the sample through a purification cartridge such as a cartridge. This requires complex, specialized equipment and the time required detracts from the useful life of the radionuclides.

[0010] WO 2012 / 063028 brochure 68 The Ga eluate was treated with a buffer solution. and further treated with an acid before being added to the bifunctional molecule to form the radiotracer. The resulting solution must be passed down an anion exchange column to concentrate it. Such procedures require skilled staff and are not always available. They require complex equipment, including but not limited to: Furthermore, 67 Ga radiolabeling was first performed in citrate (which is at an acidic pH) 67 A step of reacting Ga with a chelating agent complex followed by a subsequent relaxation step again comprising two steps. This was carried out by treating with a buffer solution.

[0011] So-called "cold kits" were previously produced for use in technetium radiolabeling. These are relatively simple to use and allow for the important handling of radionuclides. However, 68 In contrast to Ga, technetium typically has a It is obtained directly from the generator at near neutral pH, ~8, usually around 7.

[0012] Applicant has developed certain types of radionuclides which may optionally be attached to a targeting group. Species chelators are either specifically at low pH or neutral pH, making them radioactive. Can be formulated in a way that allows it to be used directly with gallium solutions such as nuclide eluates As a result, the composition provides a long-lasting therapeutic effect that can be used in clinical settings, such as in hospitals. To provide a portable, versatile, and easy-to-use "kit" can be done. Summary of the Invention [Means for solving the problem]

[0013] According to a first aspect of the present invention, a method for radiotherapy or for medical imaging is provided. 1. A method for preparing a complex containing a radioactive isotope of gallium, said method comprising the steps of: The gallium radioisotope solution obtained directly from the condenser was mixed with a pharmaceutically acceptable buffer. , optionally in an amount sufficient to raise the pH to a level in the range of 3 to 8, and a basic reagent acceptable therefor, wherein the composition has a pH of A chelating agent capable of chelating radioactive gallium within a range and at moderate temperatures is provided. and wherein said chelating agent is optionally linked to a biological targeting agent. The law is provided. [Brief explanation of the drawings]

[0014] [Figure 1] FIG. 1 is a graph showing the results of a comparison of the chelation efficiency of a range of chelating agents using the method of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0015] Applicant has demonstrated that effective gallium radiolabeling can be achieved by using gallium solutions, particularly those containing gallium at a pH generally below 2. at a pH of, for example, 1 68 Highly acidic solutions obtained from Ga radionuclide generators, etc. It has been found that this can be achieved directly by contact with an acidic solution of gallium. As a result, the gallium labeling procedure can be carried out using, for example, an ion exchange column. or simplification by avoiding additional steps such as membrane-based purification or concentration steps In certain embodiments, the gallium radioisotope solution may be 68 In the case of a Ga solution, it is not necessary to subject it to an initial concentration step at all, and the solution There is no need to pass it through an ion exchange medium.

[0016] A suitable gallium-68 gemstone can be used to provide a gallium radioisotope solution. The injector is Eckert & Ziegler's GallaPharm 9,1 RE-Elite Galli Eo™ and Parsisotope Gall Includes uGEN.

[0017] The method also involves the use of gallium citrate, which may be produced from a cyclotron. 67 Ga salt The resulting radiolabeled product can be used for radiotherapy or molecular imaging. It may be of sufficient purity that it can be used directly in medical treatments such as rinsing. The cyclotron product is a "gallium dimer" as required by the method of the present invention. The radioactive gallium solution obtained directly from the generator.

[0018] The acidic solution, such as the elution solution, may contain pharmaceutically acceptable buffers and chelating agents, and also If desired or necessary, a pharmaceutically acceptable basic agent may be added to the composition. As used herein, the term "basic reagent" refers to a compound that is contacted with an acidic substance. "Anti-inflammatory" refers to a compound that can produce a neutralizing effect when combined with other compounds.

[0019] Therefore, the chelating agent may be combined with a pharmaceutically acceptable buffer as a "premix." and a commercially acceptable basic agent. The acidic gallium solution is added directly to the premix composition so that provides a simple "single step" procedure.

[0020] Chelating agents for radionuclides are preferably used at moderate temperatures, for example, 10-30°C. at ambient temperature, at moderate pH, e.g., 3-8, and at low concentrations (e.g., 1-10 μM). Any chelate that is effective and achieves acceptable yields in a short time (e.g., 1-5 minutes) In this case, acceptable yield of the complex is achieved by using a small amount of the administered radiolabel. It may be at least 60%, for example at least 70%, 80%, 90% or 95%.

[0021] Chelation is carried out at moderate temperatures, particularly ambient temperatures, so that heating steps or stages can be avoided. This can be achieved in a short time, thus simplifying the procedure and improving the radioactivity of gallium. This type is effective at neutral pH as well as low pH. Various types of universal chelating agents are known in the art.

[0022] For example, suitable chelating agents include HBED, DFO, DTPA, DOTA, TRAP, NOTA, NOPO, NODAGA, MPO, 6SS, B6SS, PLED, TAME, NTP, and BAPEN.

[0023] In particular, the radionuclide chelator is of formula (I) [ka] or a salt thereof, wherein one of X and Y is C=O and the other is NR; and p is independently selected from 0 to 6; R 1 can chelate radionuclides is a chelating group capable of [ka] (In the formula, R, R 2 , R 3 and R 4 are independently hydrogen or an optionally substituted C ~7 alkyl group) Selected from; and Z is hydrogen or a group of formula -B'-H, -B'-A, or a group -B'-A * -T Here, T is a targeting group capable of binding to a target of interest in a subject; A is a reactive group that allows coupling to group T, A * is the reacted reactive group A; B' is a linker group for connecting the chelating group to the reactive group A and has the formula: [ka] wherein each Q independently represents -NR 5 -, -C(O)NR 5 -, -C(O)O, -NR 5 C (O)NR 5 -, -NR 5 C(S)NR 5 - and -O-, and each R 5 are independently hydrogen or optionally substituted C 1~7 is an alkyl group, and each q and and s are independently selected from 0 to 6, and each r is independently selected from 1 to 6. represented by] is a compound of

[0024] The chelating agents of formula (I) are highly efficient in the pH range of 3 to 8 and exhibit short-term activity at low concentrations. Able to chelate radionuclides, such as gallium radionuclides, at moderate temperatures and for a period of time This general type of chelating agent only works at low pH, and is therefore suitable for pharmaceutical applications. This provides a useful advance over many of the previously known chelating agents, resulting in compositions that are This type of chelating agent, together with a neutralizing alkaline salt and a buffering agent, is particularly suitable for use in a single unit. By combining them from the beginning in a tally composition, the applicant believes that the composition obtained from a cyclotron without the need for complex preparation or purification steps; 67 G a salt, e.g., citric acid 67 Gallium solutions with an acidic pH range, such as solutions of Ga and even at low pH, e.g., below 2. 68 Ga radionuclide It has been found that the eluate from the condenser can be used directly. It simplifies the process and allows for minimal manipulation, especially for use with gallium radionuclides, by providing a "cold This gives the possibility of forming a "net".

[0025] In certain embodiments, the reagents used in the method (chelating agents, buffers and basic reagents) ) in solid form, especially in lyophilized or freeze-dried form. This means that they can be used at any time to produce radiotracers in situ. This allows for the formation of a stable mixture that can be stored or transported in a ready state. In an embodiment, the buffer and basic components are added to the eluate from the gallium radionuclide generator. may be added thereto. The solution may then simply be added to a second vial or container containing the solid chelating agent. In another particular embodiment, all of the reagents are combined in a single unitary composition. Preferably, the unitary composition contains sufficient chelating agent for a single imaging run. In this case, the generator is divided into units that contain the unitary composition. The compound may be eluted directly into a container such as a vial.

[0026] Pharmaceutically acceptable buffers and, if necessary, any basic reagents used in the method. The amount of medicine is 68 Addition of acidic gallium solutions, such as eluates from Ga radionuclide generators The pH of the mixture formed during addition is adjusted to a pharmaceutically acceptable level, e.g., 3 to 8, e.g., , 4 to 7, such as 5.5 to 7, and especially 6 to 7.5 or pH 6.0 to 7.0 upon reconstitution. and a chelating agent chelates the gallium radionuclide. Applicant has determined that the amount of oxidative stress should be adequate to maintain the concentration of oxidative stress at a level that would be effective in preventing the release of oxidative stress. Under these circumstances, we found that the activity of the chelating agent was not significantly reduced by direct exposure to low pH. Furthermore, a problem previously encountered, particularly in relation to handling gallium solutions, No undesired precipitation of gallium occurs as a result of exposure to high pH. Compositions in this pH range can be applied directly to the patient without the undue discomfort caused by high acidity. can be administered.

[0027] The amount of chelating agent used in the method and so present in the composition is determined by the The exact nature of the chelate, the nature of the radionuclide to be chelated, and the treatment or The results will depend heavily on factors such as the nature of the imaging process. Typically, the required components in a composition for performing a single therapeutic treatment or imaging procedure are The amount of the chelating agent used is in the range of 0.1 to 10 μmol. manufactured for lyophilization to form a formulation or after reconstitution for administration The concentration of the chelating agent is preferably greater than 5 μM, for example, 10 to 100 μM.

[0028] Suitable pharmaceutically acceptable buffers include inorganic and organic buffers. Examples include sodium phosphate, dibasic sodium phosphate, potassium phosphate, and phosphate buffers, such as ammonium phosphate; bicarbonate or carbonate buffers; disodium succinate Succinic acid buffers such as sodium thorium hexahydrate; boric acid buffers such as sodium borate; cacodylate citrate buffers such as sodium citrate or potassium citrate; Examples of organic buffers include sodium, zinc chloride, and zwitterionic buffers. Tris(hydroxymethyl)aminomethane (TRIS) buffers, e.g., Tris HCl, T Tris EDTA, Tris acetate, Tris phosphate or Tris glycine such as morpholinepropanesulfonic acid (MOPS), and N-(2-hydroxyethyl ) Piperazine-N'(2-ethanesulfonic acid) (HEPES), dextrose, lactose ammonium, sodium or potassium acetate, etc. In certain embodiments, the buffer is other than acetate buffer, and acetate Other than sodium buffers.

[0029] Preferably, the buffer is a phosphate buffer, such as a sodium phosphate buffer. , may contain one or more phosphates, particularly monobasic and dibasic sodium phosphates. For example, a suitable buffering agent may be sodium phosphate monobasic anhydrous, and dibasic sodium phosphate heptahydrate in a ratio of about 1.5:1 to 2.5:1.

[0030] The total amount of buffer present will depend on the specific characteristics of the buffer and the characteristics of the complex and the procedure to be followed. The availability of molecular imaging modalities will depend on factors such as the specific molecular imaging modality being used. In the present invention, the buffer is present in the dry composition in an amount of 5 to 95 mole percent. Liquid compositions, e.g., those prepared for lyophilization to form solid compositions, or After reconstitution for administration, the concentration of the buffer reagent is preferably 0.01 to 0.6M, e.g. For example, it is in the range of 0.1 to 0.5 M, for example, about 0.2 M (20 mM).

[0031] In some embodiments, the buffer is a particularly "strong" buffer, such as ammonium acetate. In some cases, it may not be necessary to include a pharmaceutically acceptable basic agent. However, in certain embodiments, the pharmaceutically acceptable basic reagent is used to neutralize the eluate. Suitable pharmaceutically acceptable basic agents include sodium hydroxide, sodium phosphate, sodium phosphate phosphate, sodium phosphate phosphate phosphate, sodium phosphate phosphate phosphate phosphate. Alkali or alkaline, such as sodium, potassium, calcium or magnesium Alkaline salts such as hydroxides, carbonates, bicarbonates or oxides of earth metals, or Suitable reagents include ammonium salts or basic organic reagents. For example, suitable reagents include sodium hydroxide, ammonium hydroxide, potassium hydroxide, magnesium oxide, calcium carbonate, magnesium carbonate Magnesium, magnesium aluminum silicate, sodium carbonate, sodium bicarbonate, In particular, the drug may be selected from the group consisting of ethanolamine, thiamin mononitrate ... A physiologically acceptable basic agent is an alkali metal hydroxide, particularly sodium hydroxide or In an alternative embodiment, the basic agent is sodium bicarbonate. It is thorium.

[0032] The amount of pharmaceutically acceptable basic reagent present in the composition will depend on the exact nature of the reagent, the kit and thus the particular radioactive It will depend heavily on factors such as the pH of the eluate from the nuclide generator. Typically, such reagents are used in a composition for use in a single therapeutic or imaging procedure. The amount of the drug is 0.5 to 0.75 millimoles. Therefore, the liquid composition, e.g., the solid composition, Prepared for lyophilization to form a composition or after reconstitution for administration In this case, the concentration of the basic reagent is preferably 0.01 to 0.6M, for example 0.1 to 0.15M. It's in range.

[0033] In certain embodiments, the chelator of formula (I) comprises a targeting moiety as defined above. May or has the potential to become bound to T.

[0034] The complexes used in the compositions of the present invention are preferably covalently bonded, in particular as described above, and a biological targeting agent linked to a chelating agent, wherein the compound has the formula (I): The compound contains the group "T", but otherwise the biological targeting agent is not bound by other means. In particular, the biological targeting The tropic agent is covalently attached to a chelating agent, the chelating agent having the formula (II) B'-A * -T) [ka] or a salt thereof; * , B', X, Y, R 1 , m and p are as defined in

[0035] In another embodiment, the chelator is a compound with which a targeting group can be reacted. Therefore, formula (III) [ka] or a salt thereof; wherein T, A, B', X, Y, R 1 , m and p are As defined.

[0036] Particularly preferred examples of formula (I) are those described in WO 2012 / 063028 It is listed.

[0037] In certain embodiments, R 1 is a group of sub-formula (i) or (ii) as defined above , such as a group of sub-formula (i):

[0038] Preferably, R 3 and R 4 is hydrogen or a lower C 1~4 Selected from alkyl groups In certain embodiments, R 3 is hydrogen. In another particular embodiment, R 4 Hame It's chill.

[0039] In certain embodiments, each X, Y, m, p, Q, s, r, and q are similar.

[0040] In certain embodiments, X is C(O) and Y is NR. Suitably, R is hydrogen or or C 1~4 It is an alkyl group, especially hydrogen.

[0041] In certain embodiments, p is 1. In another particular embodiment, m is 2.

[0042] In certain embodiments, q is 0.

[0043] Preferably, within group B, Q is a group —C(O)NR 5 In particular, R 5 is hydrogen, such as hydrogen and C 1~4 The alkyl group is selected from the group consisting of:

[0044] Suitable biological targeting moieties T in formula (I) for use in the compositions of the invention are It will be a group that can direct molecules to different targets of interest in the biological system in question. Generally, these are therefore molecules in which the targeting moiety and the target of interest are specifically linked to each other. To have specificity, they will form a "specific binding pair" with the target of interest, and Under normal conditions, they bind to each other rather than to other molecules. Examples of conjugation pairs are well known in the art, e.g., receptors and ligands, enzymes, etc. and substrates, as well as immunoglobulins such as antibodies and antigens. The tropic moiety T may be a peptide, protein or other biomolecule, such as an aptamer, or The class of compounds of interest may also be small molecule ligands that bind to specific in vivo molecular targets. The targets of the drug include ligands or receptors expressed on diseased cells or tissues. may be a transporter, a cell surface antigen associated with a disease state, or a tumor marker, e.g., a cancer-specific marker. or tissue-specific markers.

[0045] In certain embodiments, the targeting moiety T is a target of a prostate specific membrane antigen (PSMA), such as These ligands target cancer-specific markers. A-11(Eder M. et al.,Bioconjugate Chem.2012,688 ) is included.

[0046] Alternatively, the targeting moiety T can target C cells, such as cells of the myelomonocytic lineage and leukemia cells. antibodies, such as anti-CD33 antibodies, for imaging cancer cells that express CD33; or a binding fragment thereof (Emberson et al., J. Immunol. Met. hods.305(2):135-51, 2005) or glycoproteins Binding to the glycoprotein carcinoembryonic antigen (CEA), a member of this family of proteins The antibodies that can be produced are found in colon cancer cells, stomach cancer cells, pancreatic cancer cells, lung cancer cells, and thyroid medullary cancer cells. on cancer cells and breast cancer cells, as well as anti-PSMA antibodies and their binding fragments Other suitable antibodies may exhibit affinity for cell adhesion molecules. These include macrophages, macrophages, and macrophages. It contains a monoclonal antibody, SER4, which binds to the phage adhesion molecule, sialoadhesin. Sialoadhesin is found on the surface of macrophages, e.g., in large amounts in the spleen, liver, It is found on macrophages in lymph nodes, bone marrow, colon and lungs.

[0047] Further examples of suitable T groups include T-terminal amino acids such as TMP-2, TMP-3, TMP-4, TMP-5, TMP-6, TMP-7, TMP-8, TMP-9, TMP-10, TMP-11, TMP-12, TMP-13, TMP-14, TMP-15, TMP-16, TMP-17, TMP-18, TMP-19, TMP-20, TMP-21, T (TIMP), which indicates that the expression of metalloproteinases is involved in the metastatic process. This allows imaging matrices of metalloproteinase expression (Gier See Sing et al., Bioconjug Chem. 12(6):964-71, 2001. In yet a further embodiment, the targeting moiety T is a target of complement receptor 2 ( CR2). Yet further examples include tumor, atherosclerotic undergoing angiogenesis, as commonly seen in atherosclerotic plaques, and The infarct derivatives can be linked to the bifunctional compounds by a suitable reactive group A. It is highly expressed in the endothelium of repairing diseased tissues such as damaged myocardium [alpha][ni]. Affinity for [beta]3 integrin or peptide sequence: arginine -glycine-aspartic acid (RGD). Other T groups are used, e.g., carcinoids, Somatosin is highly expressed on the surface of cancer cells in medullary thyroid carcinoma and other neuroendocrine tumors Somatostatin peptide octreotide or related compounds with affinity for the somatostatin receptor It may also include related analogs.

[0048] In another embodiment, the targeting group T may be used to target the resulting complex to apoptosis or cytotoxicity. Binding to phosphatidylserine (PS) for use in cell death imaging studies Examples of such polypeptides include Ann Examples include exin (annexin) V and the C2 domain of synaptotagmin. Polypeptides containing one or more C2 domains are well known in the art. Some polypeptides have only one C2 domain, while others have two or more. These domains generally have C2 domains at the end of their names (alphabetical order: It is represented by adding a letter (e.g., C2A, C2B, etc.) to the For proteins containing a C2 domain, the domain is simply referred to as the C2 domain Specific examples include the C2A domain of rat synaptotagmin I or the C2A domain of synaptotagmin I from other species. Examples of proteins containing C2 domains include the C2A domain of putotagmin. Examples include synaptotagmin 1-13, a serine / threonine kinase protein Kinase C family members, phospholipase A2, phospholipase 51, factors V and VII Cofactors in the coagulation cascade, such as I, and members of the copine family Human synaptotagmins include, but are not limited to, synaptotagmin Includes 1-7, 12 and 13.

[0049] Other suitable targeting moieties T include bombesin, gastrin, or VCAM targeting moieties. It is a peptide.

[0050] The targeting moiety T may be linked to a suitable linker group A * The chelating agent molecule is bound to the I) to form a compound of formula I. * the nature of which depends on the nature of the targeting moiety T, This may be determined using conventional chemistry.

[0051] Alternatively, they may be compounds such as bisphosphonate derivatives that target bone, particularly osteosarcoma. The calcium chelating group may be

[0052] In an alternative embodiment, the chelator may not contain a targeting group, but may be used to inhibit renal function. As a metal chelator for general radiochemical monitoring, such as in radioisotope monitoring Simply work.

[0053] Typically, the linker group A * is, in certain embodiments, a protein-reactive functional group The protein reactive group is formed from reactive group A. The protein reactive group is formed from a protein or modified protein. Alternatively, it may react with a peptide or other vehicle derivatized for this purpose. Alternatively, the protein reactive group A may be a maleimide group, an alkyl or aryl isothiocyanate group, or isothiocyanate groups such as aldehydes, esters, or alkynes, azides, and amides alkene, hydrazine, hydrazine derivatives, alkoxyamine, alkoxyamine derivatives, "Click" reagents such as aminoxy, thiol groups, etc. Amido, isothiocyanate, aldehyde and ester groups are peptide thiol- or Reacts efficiently with amine-containing residues (cysteine, lysine), so conjugation is easily achieved Other bioorthogonal functional groups can be added by converting them to alkynes, azides, alkenes, and hydroxyls. engineered peptides for coupling with hydroxyl, aminoxy, or thiol groups. It can be incorporated into amides and proteins.

[0054] As used herein, the term "alkyl" means an alkyl group having 1 to 10 carbon atoms, unless otherwise specified. It preferably means a linear or branched group containing 1 to 7 carbon atoms. "Aromatic group" refers to an aromatic group, including, for example, a phenyl group, optionally linked to an alkyl group. Taste.

[0055] The compound of formula (I) is described in WO 2012 / 063028. The compounds can be prepared using methods known in the art.

[0056] For example, compounds of formula (II) above may generally be prepared by replacing group A with a targeting moiety T or a chiral moiety. This is achieved by linking the molecule to either the ionizing agent or the ionizing agent molecule, which then connects to these other molecules. Thus, for example, they are reacted with the compound of formula (III) above. , formula (IV) TH (IV) where T is as defined above. Suitable reaction conditions depend on the exact nature of the groups A, T, etc. The exact nature of the compound will depend on factors such as the compound's molecular weight and specific characteristics, and will be determinable by a skilled chemist.

[0057] The compounds of formula (III) can be prepared by themselves, for example as disclosed in WO 2012 / 063028. As shown in Example 5 of FRET, a compound of formula (V) [ka] with a reactive group such as a maleimide group.

[0058] Certain compounds of formula (V) where B is the group HN(CH)C(O)NH are , Zhou T. et al., J. Med Chem. 2006, 49, 4171-4182 (See compound (I) in the scheme below.) This compound has the formula [ka] CP256 Tris(hydroxypyridinone) chelating agent C (also known as THP) It is a derivative of P256.

[0059] This is due to the fact that, as described in WO 2012 / 063028, It may be derivatized to form a compound of formula (III) above, where A is a maleimide group. Thus, specific compounds of formula (III) are as follows: [ka] of formula YM103 or a salt thereof.

[0060] An alternative compound of formula III is a compound of formula (I) where A is capable of binding to a primary amine. It is a compound containing an isothiocyanate group.

[0061] An example of such a compound is shown as 1 and H3THP 2 Compounds of or salts thereof. [ka]

[0062] These compounds can be synthesized, for example, by the following reaction scheme: [ka] The same as that described in International Publication No. 2012 / 063028 as shown in It can be prepared using methods similar to those described above.

[0063] In the above scheme, compounds such as those described in Zhou et al. (see above) (1 ) is reacted with triethylamine and carbon disulfide in ethanol to give, upon addition of water, This gives a precipitated dithiocarbamate intermediate (Munch et al., Tetrahedron Letters (2008), 49, 3117. The precipitated intermediate is carbon disulfide / ethanol. The mixture was resuspended in a solution of di-tert-butyl dicarbonate and a catalytic amount of 4-dimethylaminobenzoate. Addition of methylaminopyridine resulted in the formation of (2). Subsequent removal of the benzyl group followed by the addition of trifluoroethanol yields H3THP 1 Also This was purified using reverse-phase semi-preparative HPLC to give the product as a trifluoroacetate salt. And give.

[0064] H3THP 2 To synthesize A solution of (1) was added with diisopropylethylamine and then mixed with reversed-phase semi-preparative HPMC. Isolation of (3) by LC follows. Similar to (2), the benzyl group of (3) can be purified by HPLC using HCl in DCM. BCl3, followed by the addition of methanol, H3THP 2 This gives the chloride salt of

[0065] Preferred salts are pharmaceutically acceptable salts such as halide salts, especially chloride salts. Such compounds are at room temperature and physiological pH. 68 Fast radiolabeling with Ga (less than 5 min) It was found that such a pH 68 Material directly from the Ga generator This can be achieved with the compositions of the present invention using:

[0066] The compositions of the present invention may be prepared in the presence of excipients or pharmaceutically acceptable carriers as would be understood in the art. Carriers or fillers, as well as stabilizers, antimicrobials, cryoprotectants, antioxidants, free radicals Calculants, solubilizers, tonicifying agents, surfactants And, it may further contain reagents such as collapse temperature adjusters used in lyophilization.

[0067] Suitable fillers for use in the present formulation include, for example, mannitol, lactose, Sucrose, trehalose, sorbitol, glucose, or raffinose Sugars or amino acids such as arginine, glycine or histidine, as well as dextromethorphan. Polymers such as tolan or polyethylene glycol (PEG) are included.

[0068] Suitable free radical scavengers are self-radicalizing agents such as ascorbic acid or gentisic acid. Amount of free radical scavenger that may be added to the composition will depend on factors such as the nature of the capture agent used and the nature of the composition. In one embodiment, the kit may contain 1-4% w / w of a free radical scavenger. stomach.

[0069] Tonicity agents include, for example, sodium chloride, sucrose, mannitol, or dextromethorphan. The source may be selected from:

[0070] Antibacterial agents include benzyl alcohol, phenol, m-cresol, methylparaben, or It may be selected from ethylparaben.

[0071] Surfactants may include polysorbates, such as polysorbate 80.

[0072] Examples of the collapse temperature adjuster include dextran, hydroxyethyl starch, and Ficoll. It may be selected from Ficoll or Gelatin.

[0073] In certain embodiments, the composition is prepared according to the method of Belgian Patent No. 1021191, International Patent No. International Publication No. 2016030103 Brochure or International Publication No. 2016030104 It does not contain any agents that may inhibit metals other than gallium, as described in FRET. Such agents are sugars, such as monosaccharides, disaccharides or polysaccharides and their derivatives. Specific examples are glucose, fructose, β-cyclodextrin, mannose and Applicants have found that such agents are not required in the compositions of the present invention. I found that.

[0074] These compositions may be in the form of kits for use in medical imaging procedures, Such kits form a further aspect of the present invention. A kit for use in the method as described above, said kit comprising a pharmaceutically acceptable carrier. and optionally a pharmaceutically acceptable basic agent, within a pH range of 3 to 8. and a chelating agent capable of chelating radioactive gallium at moderate temperatures (the chelating agent The targeting agent includes a composition comprising: Kits are further provided, in which the chelating agent is in admixture with the buffer composition. Although they may be provided in the form of a solution, especially a sterile solution, the components of the kit are preferably , in solid form, in particular in lyophilized or freeze-dried form. , a sample held in a container sufficient to perform one or more molecular imaging techniques. The container is a sterile, sealed container that may be filled with an inert atmosphere such as nitrogen gas. Such kits may further include elements such as instructions for use and outer packaging. or in situ, using a supply of gallium radiolabel present in a suitable generator. It may be supplied to a hospital or clinic for reconstitution.

[0075] Certain compositions used in these kits are novel and are further embodiments of the present invention. The present invention therefore provides a unit for use in the method described above. A pharmaceutical composition, the composition comprising: (i) a biologically active ingredient at a pH of 3 to 8 and at a moderate temperature; Chelating a gallium radionuclide, optionally linked to a specific targeting agent and (ii) a chelating agent capable of reacting with the chelating agent; and (ii) a pharmaceutically acceptable buffer; and optionally i) a pharmaceutically acceptable basic reagent, wherein (ii) and optionally (i i) In the case where the eluate directly from the gallium generator is added to the composition, Further provided is a composition present in an amount sufficient to provide a pH in the range of 3-8.

[0076] In particular, the gallium radionuclide chelator is a compound of formula (I) as described above. The composition may be in a solution, e.g., a sterile solution, but is preferably in solid form, e.g., It is in lyophilized or freeze-dried form.

[0077] The composition of the present invention is obtained after the addition of an acidic gallium solution obtained directly from a gallium generator. In solution, it has a pH in the range of 3 to 8. It is preferably and a pharmaceutically acceptable buffer, also as described above. and a basic reagent that is tolerable thereto.

[0078] In a further aspect, the present invention provides a method for producing the composition of the present invention as described above. The method further comprises the step of: adding a chelating agent as defined above to a suitable amount of a pharmaceutically acceptable salt thereof; mixing with a basic buffer and optionally a pharmaceutically acceptable basic reagent; and optionally freeze-drying the resulting mixture.

[0079] In certain embodiments, the composition comprises mixing together the components as described above in an aqueous solution. The solution is preferably prepared by adding a filter as described above, if necessary. Formula (I) at a concentration of more than 5 μM, e.g., 10-100 μM, together with hydroxybenzoates and other excipients. ) chelating agent, base at a concentration of 0.1 to 0.6M, and and a buffering agent. The composition is then preferably dried using a method known in the art to produce a dry composition. The mixture is subjected to a freeze-drying procedure as can be seen in the art.

[0080] The amount of composition subjected to the freeze-drying procedure may be adjusted to suit one or two therapeutic procedures or imaging procedures. It may be sufficient to generate something sufficient for the mapping operation. It may be preferable to lyophilize the composition in vials, particularly glass vials. Alternatively, if a larger amount of the composition is subjected to drying, the dried composition may be added to the individual doses. It may then be divided into dosage units.

[0081] Once prepared in this manner, the composition is then in situ radiolabeled from a radiolabel generator. et al. 68 For distribution, the solution is ready to be reconstituted with an acidic gallium solution, such as Ga eluate. They may be packaged and stored for storage.

[0082] In such a generator, 68 The Ga radionuclide is fed onto the column. for use in imaging methods 68 To obtain Ga radionuclides, acids, especially At a concentration of 0.05M to 1M, for example 0.05 to 0.6M HCl, in particular about 0.1M It is eluted with inorganic acids such as hydrochloric acid.

[0083] may be used in molecular imaging or therapy, 67 Ga radionuclides are generally cyclohexane Manufactured by the Thoron procedure and citric acid 67 It is supplied in the form of an acid salt such as Ga. may be used as the acidic gallium solution in the method of the present invention.

[0084] According to the present invention, the obtained product is obtained by changing the pH of the eluate to a base and a hydroxyl group simultaneously with the labeling process. and buffering agents, so that physiological The method is rapid and requires few handling steps. This allows for minimal radiation exposure to the operator and minimal opportunity for microbial contamination. with minimal need for complex and costly equipment and Ensure that the reagent has a good useful life before it is spoiled.

[0085] The amount of eluent added to the reagent depends on the exact nature of the eluent and its composition, as well as the imaging method. the required amounts of reagents required for the composition, the size and characteristics of the patient to whom the composition is to be administered, etc. However, typically, the amount will vary depending on factors such as: 5 ml of dissolution fluid will be added to produce the appropriate dosage unit.

[0086] If desired, the dry composition of the present invention may be diluted with sterile water or saline prior to the addition of the elution fluid. In certain embodiments, the eluate is added directly to the dry reagents. .

[0087] In a further aspect, the present invention provides a method for the treatment of cancer using molecular imaging or radionuclide therapy. The present invention provides a radiolabeled product obtained by the method as described above. Electron imaging techniques are well known in the art and include PET and SPECT techniques, as well as X-ray computed tomography (CT) and Cerenkov luminescence imaging (CLI) Includes:

[0088] Thus, in a further aspect, the present invention provides a method for obtaining a molecular image of a patient, said method comprising: carrying out a process to produce a radiolabeled product as described above; administering the product to a patient in need thereof and using molecular imaging techniques. and monitoring the results.

[0089] Yet a further aspect is a method of treating a patient with a radionuclide, said method comprising the steps of: As described 67 conducting a process to produce a Ga radiolabeled product; A method is provided which comprises administering this product to a patient in need thereof.

[0090] The amount of radiolabeled product administered will depend on the patient and the targeting moiety in the composition. The nature of the organ or tissue being imaged, the radiolabel and the specific imaging technique or techniques used The exact dosage will vary depending on factors such as the nature of the treatment and the type of medication. will be determined in accordance with

[0091] The present invention therefore provides gallium, in particular 68 Used in a wide range of clinical situations where Ga is utilized The physiologically acceptable product provides an effective "cold kit" for the preparation of can be produced rapidly and easily, so the usable half-life of the radiolabel is The level of labeling using such chelating agents, in particular the compounds of formula (I), The radiochemical purity is typically greater than 95% and is therefore particularly effective, requiring significant purification. The procedure can be avoided.

[0092] The invention will now be particularly described by way of example with reference to the accompanying drawings, in which: FIG. 1 shows the results of a comparison of the chelation efficiency of a range of chelating agents using the method of the present invention. This is a graph.

[0093] However, it will be apparent to one skilled in the art that specific details are not required to practice the present invention. It will be apparent that the following descriptions of specific embodiments of the present invention are presented for purposes of illustration and description. They are presented for the purpose of being exhaustive of the invention or for the purposes of the disclosed techniques. It is not intended to limit the invention to the precise form. Many modifications and variations are possible in light of the above teachings. The embodiments are presented to best explain the principles of the invention and its practical application. and thereby allowing others skilled in the art to easily and conveniently use the present invention and various modifications thereof. Allows the embodiments to be best utilized as suited to the particular use envisaged. are shown and described for the purpose. [Example]

[0094] Example 1 68 Preparation of Ga-labeling reagents Various concentrations of pharmaceutically acceptable buffer (sodium phosphate) were added as shown in Table 1 below. A chelate containing a pharmaceutically acceptable base reagent (sodium hydroxide) and a buffering agent. A range of compositions containing the anti-inflammatory drug CP256 were prepared. The mixtures were lyophilized under vacuum overnight. .

[0095] Eckert and Zeigler 68 The Ga generator was dissolved in 0.1M HCl. The eluate was extracted to produce 5 ml of eluate containing 300 MBq per elution. was added to each of the compositions at room temperature.

[0096] The pH of the resulting solution was measured. The % radiolabeling of CP256(THP) was determined by the T Further investigation was carried out using LC, and the results are also shown in Table 1 below.

[0097] [Table 1]

[0098] The results show that radiolabeled CP256 was obtained with high efficiency in 2 minutes. The purity of the gallium may, in some cases, require further purification of the gallium before administration to patients. It would mean that you don't want to.

[0099] Example 2 68 Preparation of Ga-labeling reagents The methodology of Example 1 was repeated using 0.13 M sodium bicarbonate and 0.1 M phosphate buffer (PBS ) and a wide range of CP256 concentrations as listed in the table below. High efficiency of labeling was observed with concentrations of chiral amine as exemplified in Tables 2 and 2a. This was achieved with respect to the rate agent.

[0100] [Table 2]

[0101] [Table 3]

[0102] Example 3 Comparison of radiolabeling using different chelating agents The method of Example 1 was carried out using a range of different chelating agents (DOTA, NOTA, T The experiment was repeated using phosphate buffered saline (RAP, NOTP, HBED, DFO and THP). The amount of reagent and sodium hydroxide was adjusted to achieve a pH of either 4 or 7 upon addition of 0.1M eluent. The solution was incubated at room temperature for 10 minutes.

[0103] The results at pH 7 are shown in Figure 1. Acceptable labeling efficiencies of over 95% were observed for THP and All other chelators were found to be 95% CI at pH 7.0. Furthermore, most other chelator concentrations resulted in >90% labeling. had to be very high to achieve.

[0104] Example 4 Freeze-Drying Kit Prepared as described above and bound to the PSMA targeting agent (30 nmoles). CP256 (THP) (40 μg), sodium bicarbonate (42 mg), and monosodium chloride Sodium phosphate dibasic anhydrous (8.2 mg) and sodium phosphate dibasic heptahydrate (8.5 mg) A vial containing a lyophilized reagent mixture containing 10 mg of lyophilized acetone (Eckert and Zeigler 68 0.1M HCl eluate (5 ml) and can be used for therapy or molecular imaging, pH 6. Solutions with pH values ​​of 5 to 7.0 could be produced.

[0105] Example 5 Alternative freeze-dried kits Frozen ice cream as described in Example 4, but also containing 1-2 mg of ascorbic acid Vials containing the dry reagent mixture may also be prepared. This kit also includes Eckert and and Zeigler 68 The 0.1M HCl eluate (5 ml) obtained from the Ga generator ) and can be used for therapy or molecular imaging, pH 6.5- A solution of 7.0 can be produced.

Claims

1. Radioactive isotopes of gallium for use in radiotherapy or in medical imaging 1. A method for preparing a complex comprising: The resulting gallium radioisotope solution is diluted with a pharmaceutically acceptable buffer to a pH of 3-8. and optionally a pharmaceutically acceptable basic reagent in an amount sufficient to raise the level of the and a composition containing the agent, wherein the composition is within the pH range. and further comprising a chelating agent capable of chelating radioactive gallium at moderate temperatures, The method wherein said chelating agent is optionally linked to a biological targeting agent.

2. The gallium solution is an eluate obtained directly from a gallium-68 radionuclide generator. The method according to claim 1, wherein

3. 3. The method of claim 2, wherein the eluate is at a pH of less than 2.

4. The composition according to any one of claims 1 to 3, wherein the composition comprises a pharmaceutically acceptable basic agent. How to post.

5. The pharmaceutically acceptable basic agent is an alkali metal hydroxide, carbonate or bicarbonate. Salts, specifically sodium or potassium hydroxides, carbonates or bicarbonates The method according to claim 4, wherein

6. 6. The method of claim 1, wherein the chelating agent is linked to a biological targeting agent. The method described in paragraph .

7. The chelating agent is a compound represented by formula (I) 【Chemistry 1】 or a salt thereof, wherein one of X and Y is C=O and the other is NR; and p is independently selected from 0 to 6; R １ can chelate radionuclides is a chelating group capable of 【Chemistry 2】 (In the formula, R, R ２ , R ３ and R ４ are independently hydrogen or optionally substituted C １ ～７ alkyl group) Selected from: and Z is hydrogen or a group of formula -B'-H, -B'-A, or a group -B'-A ＊ -T Here, T is a targeting group capable of binding to a target of interest in a subject; A is a reactive group that allows coupling to group T, A ＊ is the reacted reactive group A; B' is a linker group for connecting the chelating group to the reactive group A and has the formula: 【Transformation 3】 wherein each Q is independently —NR ５ -, -C(O)NR ５ -, -C(O)O, -NR ５ C (O)NR ５ -, -NR ５ C(S)NR ５ - and -O-, and each R ５ are independently hydrogen or optionally substituted C １～７ is an alkyl group, and each q and and s are independently selected from 0 to 6, and each r is independently selected from 1 to 6. represented by] The method according to any one of claims 1 to 6, wherein the compound is

8. In the compound of formula (I), R １ is group (iii) as defined in claim 7 The method according to claim 7,

9. The compound of formula (I) is a compound of formula (II) 【Chemistry 4】 (In the formula, T, A ＊ , B, X, Y, R １ , m and p are as defined in claim 7 ） 9. The method according to claim 7 or 8, wherein the compound is:

10. The group A ＊ is a maleimide group or contains an isothiocyanate group. How to post.

11. The pharmaceutically acceptable buffer may be a phosphate buffer, a bicarbonate or carbonate buffer, Succinic acid buffer, boric acid buffer, cacodylic acid buffer, citrate buffer, sodium chloride, salt Zinc chloride, zwitterionic buffer, tris(hydroxymethyl)aminomethane (TRIS) buffer agent, morpholinepropanesulfonic acid (MOPS), N-(2-hydroxyethyl)piperane Di-N'(2-ethanesulfonic acid) (HEPES), dextrose, lactose, alcohol The method according to any one of claims 1 to 10, wherein the buffer is tartaric acid, arginine or acetate.

12. The chelating agent is a biological targeting ligand, such as a ligand that targets a cancer-specific marker. The method according to any one of claims 1 to 11, further comprising a desiccant.

13. 10. The method of claim 1, wherein the biological targeting agent is specific for prostate-specific membrane antigen (PSMA).

13. The method according to claim 12.

14. The acidic gallium solution ６８ By eluting the Ga radionuclide column with inorganic acid A method according to any one of claims 1 to 13 obtained.

15. 15. The method of claim 14, wherein the inorganic acid is hydrochloric acid.

16. A kit for use in the method according to any one of claims 1 to 15, comprising: The solution is radioactive in a pharmaceutically acceptable buffer within a pH range of 3-8 and at moderate temperatures. a chelating agent capable of chelating gallium (the chelating agent is a biological targeting agent); and optionally a pharmaceutically acceptable basic and a reagent, wherein the composition is When added to a solution obtained directly from Kit.

17. The kit of claim 16, wherein the composition is in liquid form.

18. 17. The method of claim 16, wherein the components of the kit are in lyophilized or freeze-dried form. Kit included.

19. The component is sufficient to perform one or more therapeutic procedures or molecular imaging procedures.

19. The kit according to any one of claims 16 to 18, wherein the components are present in an amount of 0.1 to 0.2g and are held in a container.

20. The chelating agent is a compound as defined in any one of claims 7 to 9. Item 20. The kit according to any one of Items 16 to 19.

21. A unitary composition for use in the method according to any one of claims 1 to 15. and (i) at a pH of 3-8 and at moderate temperatures, depending on the biological targeting agent. a chelating agent capable of chelating a gallium radionuclide, optionally bound to the and (ii) a pharmaceutically acceptable buffer, and optionally (iii) a pharmaceutically acceptable and a basic reagent, wherein (ii) and (iii) are gallium radionuclide gemini. When the solution obtained directly from the enerator is added to it, it produces a pH in the range of 3 to 8. The composition is present in the composition in an amount sufficient to

22. The gallium radionuclide chelator is as defined in any one of claims 7 to 10.

22. The composition of claim 21, wherein the compound is of formula (I).

23. 23. The composition of claim 21 or 22, in lyophilized or freeze-dried form. composition.

24. 24. The method according to claim 21, wherein the solution has a pH range of 3.0 to 8. The composition described above.

25. The composition according to any one of claims 21 to 24, which comprises a pharmaceutically acceptable basic agent. 。

26. The pharmaceutically acceptable basic agent is an alkali metal hydroxide, in particular sodium hydroxide. or potassium.

27. The pharmaceutically acceptable buffer may be a phosphate buffer, a bicarbonate or carbonate buffer, Succinic acid buffer, boric acid buffer, cacodylic acid buffer, citrate buffer, sodium chloride, salt Zinc chloride, zwitterionic buffer, tris(hydroxymethyl)aminomethane (TRIS) buffer agent, morpholinepropanesulfonic acid (MOPS), N-(2-hydroxyethyl)piperane Di-N'(2-ethanesulfonic acid) (HEPES), dextrose, lactose, alcohol 27. The composition of any one of claims 21 to 26, wherein the buffer is tartaric acid, arginine, or acetate. thing.

28. 2. The method of claim 1, wherein the biological targeting agent is a ligand that targets a cancer-specific marker.

28. The composition according to any one of claims 1 to 27.

29. 10. The method of claim 1, wherein the biological targeting agent is specific for prostate-specific membrane antigen (PSMA).

29. The composition described in 28.

30. The composition of any one of claims 21 to 28, further comprising a free radical scavenger.

31. A method for producing a composition according to any one of claims 21 to 30, said method comprising the steps of:

22. A chelating agent as defined in claim 21 in a suitable amount of a pharmaceutically acceptable basic buffer. and optionally also mixing with a pharmaceutically acceptable basic reagent, and optionally freeze-drying the mixture.

32. A method according to any one of claims 1 to 15 for use in molecular imaging or in therapy. The product obtained by the method described above.

33. 33. A method for obtaining a molecular image of a patient, said method comprising administering to the patient a product according to claim 32. and monitoring the results using molecular imaging techniques.

34. Patients ６７ A method of treating with a Ga radionuclide, said method comprising administering to said gallium radionuclide The nuclide ６７ 32. An effective amount of the product obtained by the method of claim 31, wherein the product is Ga. to a patient in need thereof.

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