Methods of making, formulating and treating aqueous preparations of manganese-containing coordination complexes

Abstract

A method is provided for producing an aqueous formulation of a manganese-containing coordination complex by combining a source of chloride anions and a source of a manganese-containing coordination complex in aqueous solution, either simultaneously or subsequently, providing a dianion source to the aqueous solution to form the aqueous formulation.

Description

[Technical Field]

[0001] This application claims priority to U.S. Provisional Application No. 62 / 913,704, filed October 10, 2019, which is incorporated herein by reference in its entirety.

[0002] The present invention relates generally to methods for making aqueous preparations of manganese-containing coordination complexes, such as manganese-containing pentaaza macrocycle complexes, and methods of formulation and treatment therewith.

[0003] Aqueous formulations of manganese-containing coordination complexes can be prepared for a variety of uses, including as parenteral drug formulations for the treatment of disease states. Parenteral formulations typically must have certain properties to be suitable for administration, such as a physiologically acceptable pH, the ability to maintain the solubility of the parenterally administered compound without substantially degrading the formulation, and acceptable isotonicity. Aqueous formulations for parenteral administration must also generally be sterile and preferably contain no visually discernible particles and a limited amount of particles below the visual threshold.

[0004] Examples of manganese-containing coordination complexes in aqueous formulations that can be administered for treatment include manganese-containing pentaaza macrocyclic complexes having a macrocyclic ring system corresponding to Formula A, which have been shown to be effective in treating a number of animal and cell models of human disease as well as conditions afflicting human patients. [ka] For example, in a rodent model of colitis, one such compound, GC4403, has been reported to significantly reduce colonic damage in rats subjected to an experimental model of colitis (see Cuzzocrea et al., Europ. J. Pharmacol., 432, 79-89 (2001)). [ka] GC4403 has also been reported to reduce radiation damage in both a clinically relevant hamster model of radiation-induced acute oral mucositis (Murphy et al., Clin. Can. Res., 14(13), 4292 (2008)) and lethal total body irradiation in adult mice (Thompson et al., Free Radical Res., 44(5), 529-40 (2010)). Similarly, another such compound, GC4419, has been shown to reduce VEGFr inhibitor-induced lung disease in a rat model (Tuder, et al., Am. J. Respir. Cell Mol. Biol., 29, 88-97 (2003)). Furthermore, another such compound, GC4401, has been shown to provide protective effects in animal models of septic shock (S. Cuzzocrea, et.al., Crit. Care Med., 32(1), 157 (2004)) and pancreatitis (S. Cuzzocrea, et.al., Shock, 22(3), 254-61 (2004)). [ka]

[0005] Some of these compounds have also been shown to have potent anti-inflammatory activity in vivo and prevent oxidative damage. For example, GC4403 has been reported to inhibit inflammation in a rat model of inflammation (Salvemini, et al., Science, 286, 304 (1999)) and prevent joint disease in a rat model of collagen-induced arthritis (Salvemini et al., Arthritis & Rheumatism, 44(12), 2009-2021 (2001)). Still other of these compounds, MdPAM and MnBAM, have shown in vivo activity in inhibiting colonic tissue injury and neutrophil accumulation in colonic tissue (Weiss et al., The Journal of Biological Chemistry, 271(42), 26149-26156 (1996)). Additionally, these compounds have been reported to have analgesic activity in a rat paw carrageenan hyperalgesia model and to reduce inflammation and edema; see, for example, US Pat. No. 6,180,620.

[0006] Compounds in this class have also been shown to be safe and effective in the prevention and treatment of disease in human subjects. For example, GC4419 has been shown to reduce oral mucositis in head and neck cancer patients undergoing chemoradiotherapy (Anderson, C., Phase 1 Trial of Superoxide Dismutase (SOD) Mimetic GC4419 to Reduce Chemoradiotherapy (CRT)-Induced Mucositis (OM) in Patients (pts) with Mouth or Oropharyngeal Carcinoma (OCC), Oral Mucositis Research Workshop, MASCC / ISOO Annual Meeting on Supportive Care in Cancer, Copenhagen, Denmark (June 25, 2015); Anderson, C., Phase 1b / 2a Trial of Superoxide Dismutase Mimetic GC4419 to Reduce Chemoradiotherapy-Induced Oral Mucositis in Patients with Oral Cavity or Oropharyngeal Carcinoma, Int. J. of Radiation Oncol. Biol. Phys., Vol. 100, No. 2, pages 109–110). 427-435 (2018)).

[0007] Furthermore, transition metal-containing pentaaza macrocyclic complexes corresponding to this class have shown efficacy in the treatment of various cancers. For example, some compounds corresponding to this class have been applied in combination with drugs such as paclitaxel and gemcitabine to enhance cancer therapy, such as in the treatment of colorectal cancer and lung cancer (non-small cell lung cancer) (see, e.g., U.S. Patent No. 9,198,893). The 4403 compound has also been used to treat Meth A spindle cell squamous carcinoma and RENCA renal carcinoma in vivo models (Samlowski et al., Nature Medicine, 9(6), 750-755 (2003)), and in spindle cell squamous carcinoma metastases (Samlowski et al., Madame Curie Bioscience Database (Internet), 230-249 (2006)). The 4419 compound has also been used in combination with cancer therapy, for example, in combination with cisplatin and radiation therapy, to enhance treatment in in vivo models (Sishc et al., poster for Radiation Research Society (2015)).

[0008] Thus, there is a need for improved methods of manufacturing aqueous formulations of manganese-containing coordination complexes, including formulations for parenteral administration of manganese-containing pentaaza macrocyclic complexes to treat disease states. There is also a need for improved manganese-containing coordination complexes to provide for parenteral administration and / or other treatments using such formulations while maintaining acceptable stability, isotonicity, pH, and other properties of the formulations.

[0009] Briefly, therefore, one aspect of the present invention relates to a method for preparing an aqueous formulation of a manganese-containing coordination complex comprising a manganese-containing coordination complex, a chloride anion, and a dianion, the method comprising combining a source of the manganese-containing coordination complex with a source of chloride anion in an aqueous solution, and simultaneously or subsequently providing a source of dianion to the aqueous solution to form the aqueous formulation. According to certain embodiments, the source of the manganese-containing coordination complex can comprise a manganese-containing component that includes one or more manganese that is uncoordinated or coordinated to one or more ligands other than one or more ligands of the manganese-containing coordination complex, e.g., uncomplexed manganese remaining as an impurity from the synthesis of the manganese-containing coordination complex. According to certain embodiments, the amount of chloride anion source combined with the manganese-containing coordination complex is sufficient to provide a chloride ion concentration in the aqueous formulation that exceeds the dianion concentration in the aqueous formulation.

[0010] Aspects of the present invention further relate to methods of treating a condition in a patient, comprising parenterally administering a buffered solution comprising an aqueous formulation of a manganese-containing coordination complex described herein.

[0011] A further embodiment of the invention relates to a buffered formulation for parenteral administration of a manganese-containing pentaaza macrocyclic complex, comprising a buffered aqueous solution comprising (i) a manganese-containing pentaaza macrocyclic complex at a concentration of 1 mg / mL to 50 mg / mL; (ii) sodium chloride at a concentration of 130 mM to 160 mM; and (iii) a buffering agent comprising a bicarbonate salt at a concentration sufficient to buffer the aqueous solution to a pH in the range of 7 to 10. The storage stability of the buffered formulation is such that no manganese-containing precipitate is detectable by visual inspection for 9 months after preparation of the buffered formulation.

[0012] Other objects and features of aspects of the present invention are described below. [Brief explanation of the drawings]

[0013] [Figure 1]1 shows the results of manganese measurement by ICP-MS one day after forming the aqueous formulation.

[0014] [Figure 2] A graphical representation of the results is shown in Figure 1.

[0015] [Figure 3] 1 shows the results of manganese measurement by ICP-MS 6 days after formation of the aqueous formulation.

[0016] [Figure 4] A graphical representation of the results is shown in Figure 3.

[0017] [Figure 5] Shown are photographs of MnCO3 crystals formed in an aqueous formulation after 9 months of storage, viewed with plane polarized light (bottom left) and crossed polarized light (top right) (placed in water).

[0018] [Figure 6] (A) Raman spectrum collected from the MnCO crystal of Figure 5, as well as (B) a library reference spectrum of rhodochrosite, (C) a Raman spectrum collected from a sample of MnO, and (D) a Raman spectrum compared to a library reference spectrum of eriochrosite, MnO.

[0019] Abbreviations and Definitions The following definitions and methods are provided to more particularly define the present invention and to guide those of ordinary skill in the art in the practice of the present invention. Unless otherwise noted, terms are to be understood according to conventional usage by those of ordinary skill in the art.

[0020] "Acyl" means a -COR moiety where R is alkyl, haloalkyl, optionally substituted aryl, or optionally substituted heteroaryl, as defined herein, e.g., acetyl, trifluoroacetyl, benzoyl, and the like.

[0021] "Acyloxy" means an -OCOR moiety where R is alkyl, haloalkyl, optionally substituted aryl, or optionally substituted heteroaryl, as defined herein, e.g., acetyl, trifluoroacetyl, benzoyl, and the like.

[0022] "Alkoxy" means an --OR moiety where R is alkyl as defined above, e.g., methoxy, ethoxy, propoxy, or 2-propoxy, n-butoxy, isobutoxy, or tert-butoxy, and the like.

[0023] "Alkyl" means a linear saturated monovalent hydrocarbon moiety, of one to six carbon atoms, or a branched saturated monovalent hydrocarbon moiety, of three to six carbon atoms, e.g., a C1-C6 alkyl group, such as methyl, ethyl, propyl, 2-propyl, butyl (including all isomers), pentyl (including all isomers), etc.

[0024] Furthermore, unless otherwise specified, the term "alkyl," as used herein, is intended to include both "unsubstituted alkyls" and "substituted alkyls," the latter of which refers to alkyl moieties having substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone. Indeed, unless otherwise specified, all groups recited herein are intended to include both substituted and unsubstituted options.

[0025] The term “C x-y " when used in conjunction with chemical moieties such as alkyl and aralkyl, is meant to include groups containing x to y carbons in the chain. For example, the term C x-y Alkyl refers to a substituted or unsubstituted saturated hydrocarbon group, including straight-chain and branched-chain alkyl groups, containing x to y carbons in the chain.

[0026] "Alkylene," unless otherwise indicated, means a linear saturated divalent hydrocarbon moiety of, e.g., one to six carbon atoms, or a branched saturated divalent hydrocarbon moiety of, e.g., three to six carbon atoms, e.g., methylene, ethylene, propylene, 1-methylpropylene, 2-methylpropylene, butylene, pentylene, etc.

[0027] "Alkenyl" means a linear unsaturated monovalent hydrocarbon moiety, such as of two to six carbon atoms, or a branched saturated monovalent hydrocarbon moiety, such as of three to six carbon atoms, e.g., ethenyl (vinyl), propenyl, 2-propenyl, butenyl (including all isomers), pentenyl (including all isomers), and the like.

[0028] "Alkaryl" means a monovalent moiety derived from an aryl moiety by the replacement of one or more hydrogen atoms with an alkyl group.

[0029] "Alkenylcycloalkenyl" means a monovalent moiety derived from an alkenyl moiety by the replacement of one or more hydrogen atoms with a cycloalkenyl group.

[0030] "Alkenylcycloalkyl" means a monovalent moiety derived from a cycloalkyl moiety by the replacement of one or more hydrogen atoms with an alkenyl group.

[0031] "Alkylcycloalkenyl" means a monovalent moiety derived from a cycloalkenyl moiety by the replacement of one or more hydrogen atoms with an alkyl group.

[0032] "Alkylcycloalkyl" means a monovalent moiety derived from a cycloalkyl moiety by the replacement of one or more hydrogen atoms with an alkyl group.

[0033] "Alkynyl" means an unsaturated monovalent hydrocarbon moiety, a linear unsaturated monovalent hydrocarbon moiety, such as of 2 to 6 carbon atoms, or a branched saturated monovalent hydrocarbon moiety, such as of 2 to 6 carbon atoms, e.g., ethynyl, propynyl, butynyl, isobutynyl, hexynyl, and the like.

[0034] "Alkoxy" means a monovalent moiety derived from an alkyl moiety by the replacement of one or more hydrogen atoms with hydroxy groups.

[0035] "Amino" means R a and R b are independently hydrogen, alkyl, or aryl; a R b means a group.

[0036] "Aralkyl" means a monovalent moiety derived from an alkyl moiety by the replacement of one or more hydrogen atoms with an aryl group.

[0037] "Aryl" means a monovalent monocyclic or bicyclic aromatic hydrocarbon moiety of 6 to 10 ring atoms e.g., phenyl or naphthyl.

[0038] "Ring" means a carbocyclic saturated monovalent hydrocarbon moiety of 3 to 10 carbon atoms.

[0039] "Cycloalkyl" means a cyclic saturated monovalent hydrocarbon moiety of three to ten carbon atoms, e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.

[0040] "Cycloalkylalkyl" means a monovalent moiety derived from an alkyl moiety by replacing one or more hydrogen atoms with a cycloalkyl group, such as cyclopropylmethyl, cyclobutylmethyl, cyclopentylethyl, or cyclohexylethyl.

[0041] "Cycloalkylcycloalkyl" means a monovalent moiety derived from a cycloalkyl moiety by replacing one or more hydrogen atoms with a cycloalkyl group.

[0042] "Cycloalkenyl" means a cyclic monounsaturated monovalent hydrocarbon moiety of three to ten carbon atoms, e.g., cyclopropenyl, cyclobutenyl, cyclopentenyl, or cyclohexenyl.

[0043] "Cycloalkenylalkyl" means a monovalent moiety derived from an alkyl moiety by replacing one or more hydrogen atoms with a cycloalkenyl group, such as cyclopropenylmethyl, cyclobutenylmethyl, cyclopentenylethyl, or cyclohexenylethyl.

[0044] "Ether" means a monovalent moiety derived from an alkyl moiety by the replacement of one or more hydrogen atoms with an alkoxy group.

[0045] "Halo" means fluoro, chloro, bromo or iodo, preferably fluoro or chloro.

[0046] A "heterocycle" or "heterocyclyl" is a heterocyclic ring in which one or two ring atoms are N, O, or S(O). nwhere n is an integer from 0 to 2, and the remaining ring atoms are C. The heterocyclyl ring is optionally fused to (one) aryl or heteroaryl ring, as defined herein, provided that the aryl and heteroaryl rings are monocyclic. A heterocyclyl ring fused to a monocyclic aryl or heteroaryl ring is also referred to herein as a "bicyclic heterocyclyl" ring. Furthermore, one or two ring carbon atoms in the heterocyclyl ring can be optionally substituted with a -CO- group. More specifically, the term heterocyclyl includes, but is not limited to, pyrrolidino, piperidino, homopiperidino, 2-oxopyrrolidinyl, 2-oxopiperidinyl, morpholino, piperazino, tetrahydropyranyl, thiomorpholino, and the like. When a heterocyclyl ring is unsaturated, it may contain one or two double bonds, provided that the ring is not aromatic. When a heterocyclyl group is a saturated ring and is not fused to an aryl or heteroaryl ring as described above, it is also referred to herein as a saturated monocyclic heterocyclyl.

[0047] "Heteroaryl" means a monovalent monocyclic or bicyclic aromatic moiety of 5 to 10 ring atoms in which one or more, preferably 1, 2, or 3, ring atoms are heteroatoms selected from N, O, or S, and the remaining atoms are carbon. Representative examples include, but are not limited to, pyrrolyl, pyrazolyl, thienyl, thiazolyl, imidazolyl, furanyl, indolyl, isoindolyl, oxazolyl, isoxazolyl, benzothiazolyl, benzoxazolyl, benzimidazolyl, quinolinyl, isoquinolinyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazolyl, tetrazolyl, and the like.

[0048] "Nitro" means -NO2.

[0049] "Organosulfur" means the monovalent moiety -SR, where R is hydrogen, alkyl, or aryl.

[0050] "Substituted alkyl," "substituted ring," "substituted phenyl," "substituted aryl," "substituted heterocycle," and "substituted nitrogen heterocycle" refer to an alkyl, ring, aryl, phenyl, heterocycle, or nitrogen-containing heterocycle, respectively, optionally substituted with one, two, or three substituents, e.g., substituents independently selected from alkyl, alkoxy, alkoxyalkyl, halo, hydroxy, hydroxyalkyl, or organosulfur. In general, the term "substituted" refers to any of the C 1-4 Alkyl, C 2-4 This includes groups substituted with one or more of alkenyl, halogen, alcohol and / or amine.

[0051] "Thioether" means a monovalent moiety derived from an alkyl moiety by the replacement of one or more hydrogen atoms with an --SR group, where R is alkyl.

[0052] As used herein, (i) compounds referred to in this specification and drawings as compound 401, 4401 or GC4401 are references to the same compound, (ii) compounds referred to in this specification and drawings as compound 403, 4403 or GC4403 are references to the same compound, (iii) compounds referred to in this specification and drawings as compound 419, 4419 or GC4419 are references to the same compound, and (iv) compounds referred to in this specification and drawings as compound 444, 4444 or GC4444 are references to the same compound.

[0053] Furthermore, the use of the term "consisting essentially of" in a method of treatment means that the method does not substantially include providing another therapy and / or another active agent other than the active agent specifically recited in the claim, in an amount and / or under conditions sufficient to provide treatment. Similarly, the use of the term "consisting essentially of" in reference to a kit for treatment means that the kit does not substantially include providing another therapy and / or another active agent other than the active agent specifically recited in the claim, in an amount and / or under conditions sufficient to provide treatment.

[0054] Detailed Description of the Invention In certain embodiments, aspects of the present invention relate to methods for preparing aqueous formulations of manganese-containing coordination complexes, e.g., aqueous formulations for parenteral administration of manganese-containing pentaaza macrocyclic complexes. Specifically, it has been unexpectedly discovered that careful control of aspects of the preparation method can improve the stability of such formulations. Without being bound by any particular theory, providing an appropriate protective anion in the formulation solution at critical points during preparation can minimize the formation of manganese-containing precipitates in the formulation, which could otherwise render the formulation unsuitable or undesirable for parenteral administration. Furthermore, while preparation methods are described herein for aqueous formulations intended for parenteral administration of manganese-containing pentaaza macrocyclic complexes, aspects of the present invention are not limited thereto, as similar principles are believed to apply to the use of these aqueous formulations in areas other than parenteral administration.

[0055] According to certain embodiments, aqueous formulations used for parenteral administration can be formed by combining a manganese-containing coordination complex with a salt, such as sodium chloride, and a buffer, such as sodium bicarbonate, or other buffering agent, to provide a physiologically compatible aqueous solution for administration. However, it has been unexpectedly discovered that manganese-containing coordination complexes in solution can form unwanted precipitates under certain circumstances. Specifically, without being bound by any particular theory, it is believed that if the solution does not provide sufficient protective anions to inhibit the formation of such precipitates, unwanted precipitates can form from "free" manganese or other manganese-containing impurities (described in detail below) present in trace amounts as impurities in the manganese-containing coordination complex. Thus, by providing protective anions at critical points during manufacture, the formation of unwanted precipitates can be inhibited, or even prevented, providing a sufficiently stable composition suitable for parenteral administration.

[0056] According to one embodiment, it has been unexpectedly discovered that the formation of undesired precipitates in aqueous formulations can be inhibited by providing an aqueous solution containing a manganese-containing coordination complex with chloride anions, such as chloride anions formed by dissolving a chloride-containing salt in the aqueous solution. Specifically, without being limited to any theory, it is believed that the chloride anions may provide a protective effect against any "free" manganese or other manganese-containing impurities that may be present in trace amounts as impurities in the manganese-containing coordination complex, thereby inhibiting the interaction of such "free" manganese with other anions that may be added to the solution and thus predispose to the formation of undesired precipitates. Again, without being bound by any particular theory, it is believed that anions that may form undesired precipitates with "free" manganese or other manganese compounds may be dianions, or in other words, anions each having two negative charges, as opposed to the single negative charge of the chloride anion. According to certain embodiments, the formation of undesired precipitates can be inhibited, and even substantially prevented, by providing a source of chloride anions to the aqueous solution containing the source of manganese-containing coordination complex prior to or simultaneously with the addition of the source of dianions. That is, it has been discovered that when dianions are added to the aqueous formulation, they are ideally added simultaneously with or after the protective chloride anions are added to the aqueous formulation, such that "free" manganese or other manganese-containing impurities can be substantially protected from interaction with the dianions, thereby reducing and / or eliminating the formation of precipitates.

[0057] Thus, in certain embodiments, it has been unexpectedly discovered that when bicarbonate anions are provided in an aqueous solution, the formation of undesired precipitates can be inhibited, and even prevented, by adding a chloride anion source prior to or simultaneously with the addition of bicarbonate to the aqueous solution containing the manganese-containing coordination complex. Bicarbonate anions can be provided in aqueous formulations, for example, to provide a buffer system that maintains a physiological pH of the aqueous formulation suitable for parenteral administration of the formulation. However, bicarbonate anions are also provided in aqueous formulations containing the dianion carbonate (CO3 2-), and it has been unexpectedly discovered that this dianion, without being limited to any theory, is believed to undesirably react with "free" manganese and / or other manganese-containing impurities in aqueous solution and, further, to form a precipitate with such manganese (i.e., a manganese carbonate (MnCO) precipitate) over time. Surprisingly, it has been discovered that by providing a chloride anion source prior to or simultaneously with the dianion source (e.g., bicarbonate), the formation of the unwanted precipitate is inhibited and the aqueous formulation remains stable and suitable for parenteral administration. In contrast, it has been unexpectedly observed that aqueous formulations of manganese-containing coordination complexes form an unwanted precipitate when the dianion source (e.g., bicarbonate) is provided prior to the chloride anion source.

[0058] The formation of a precipitate caused by the addition of bicarbonate to an aqueous solution containing a manganese-containing coordination complex in the absence of chloride anion is even more surprising because, following formation of the aqueous formulation, the precipitate is not immediately visually observable; instead, as described in more detail in the Examples herein, visible levels of precipitate formation can only be observed after a significant period of time has passed after preparation of the aqueous formulation, e.g., 9 months. Accordingly, it has been unexpectedly discovered that the addition of a chloride anion source prior to or simultaneously with the dianion source (e.g., bicarbonate) is important to inhibit or prevent precipitate formation caused by the interaction of the dianion with "free" manganese and / or other manganese-containing impurities, and to provide an aqueous solution that is substantially free of precipitate and therefore suitable for parenteral administration.

[0059] Without being limited to any theory, in certain embodiments, the +2 charge (Mn 2+ ) (or more) or other manganese-containing impurities are provided in solution with dianions and without any chloride anions, the manganese species may be converted to manganese by the dianion (-2) (e.g., carbonate anion CO3 2-) and has sufficient binding sites (+2) available to form the unwanted precipitate. Furthermore, again without being limited to any theory, if a sufficient amount of chloride anions with a charge of -1 are provided in solution along with "free" manganese, the manganese-containing species and chloride ions will combine to form MnCl and MnCl + (and Mn 3+ and similar species of higher oxidation states) can form in equilibrium in solution, or in other words, the equilibrium solution is neutral or has a charge of −1, thereby making Mn available for binding to the dianion, such that precipitate formation is reduced or even eliminated. 2+ Thus, when provided in sufficient amounts prior to or simultaneously with the addition of a dianion, such as carbonate anion, chloride anion can provide a protective effect.

[0060] Thus, in one aspect of the present invention, there is provided a method for preparing an aqueous formulation of a manganese-containing coordination complex, the formulation comprising the manganese-containing coordination complex, a chloride anion, and a dianion. The method generally comprises combining a source of the manganese-containing coordination complex and a source of chloride anion in an aqueous solution. The method further comprises providing a dianion source to the aqueous solution simultaneously with or after combining the chloride anion source and the manganese-containing coordination complex source to provide the aqueous formulation. That is, in some embodiments, the dianion source is combined with the manganese-containing coordination complex source only after or simultaneously with combining the chloride anion source and the manganese-containing coordination complex source. In certain aspects, the chloride anion source may thus provide a protective effect, inhibiting the formation of precipitates caused by the interaction of the dianion with trace amounts of "free" manganese or other manganese-containing impurities present in the manganese-containing coordination complex source.

[0061] In certain embodiments, the source of a manganese-containing coordination complex comprises manganese coordinated to a macrocyclic ligand. For example, in certain embodiments, the source of a manganese-containing coordination complex can comprise, among other possible macrocyclic ligands, any one selected from the group consisting of pentaaza macrocyclic ligands, tetraaza macrocyclic ligands, porphyrin macrocyclic ligands, phthalocyanine macrocyclic ligands, and crown ether macrocyclic ligands. Furthermore, in certain embodiments, the manganese-containing coordination complex comprises manganese coordinated to one or more monodentate or polydentate ligands via nitrogen atoms of one or more of the ligands. Furthermore, in certain embodiments, the manganese-containing coordination complex comprises manganese in the +2 or +3 oxidation state (Mn(II) or Mn(III)), and in certain embodiments, the manganese-containing coordination complex comprises manganese in the +2 oxidation state (Mn(II)). In certain embodiments, as described further herein below, the manganese-containing coordination complex comprises a manganese-containing pentaaza macrocycle complex, e.g., any of those further described herein below, including, among others, e.g., the macrocycle complexes designated herein as GC4419, GC4403, GC4711, and GC4702. In certain embodiments, the aqueous formulation comprises a manganese-containing coordination complex, e.g., a pentaaza macrocycle complex, at a concentration of at least 1 mg / mL, at least 3 mg / mL, at least 5 mg / mL, at least 9 mg / mL, at least 15 mg / mL, at least 18 mg / mL, and / or at least 20 mg / mL, but generally not greater than 100 mg / mL, e.g., not greater than 75 mg / mL, not greater than 50 mg / mL, not greater than 30 mg / mL, not greater than 20 mg / mL, and / or not greater than 10 mg / mL. For example, the concentration of a manganese-containing coordination complex, such as a pentaaza macrocycle complex, can range from 1 mg / mL to 50 mg / mL, such as from 5 mg / mL to 15 mg / mL, and even from 3 mg / mL to 10 mg / mL.For example, the aqueous formulation comprises at least 10 mg, at least 25 mg, at least 30 mg, at least 50 mg, at least 65 mg, at least 70 mg, at least 75 mg, at least 80 mg, at least 85 mg, at least 90 mg, at least 95 mg, at least 100 mg, at least 105 mg, at least 110 mg, at least 115 mg, and / or at least 120 mg, but generally an amount of pentaaza macrocyclic complex (e.g., GC4419) of at least 500 mg.

[0062] In certain embodiments, the source of a manganese-containing coordination complex further includes a Mn(II)-containing component that includes one or more Mn(II) atoms, either uncoordinated (i.e., "free" Mn metal not coordinated to any ligand) or coordinated to one or more ligands other than one or more ligands of the manganese-containing coordination complex. For example, the Mn(II)-containing component can include MnCl, or in certain cases, other forms of Mn(II) that are other than the manganese-containing coordination complex. Without being bound by any theory, it is believed that, as described herein, the Mn(II)-containing component can contribute to the formation of precipitates if the manufacturing process is not controlled according to the embodiments described herein. Furthermore, in certain cases, such Mn(II)-containing components can arise during the manufacture and / or synthesis of the manganese-containing coordination complex itself and can be present in the source of the manganese-containing coordination complex as relatively harmless impurities or by-products. In certain embodiments, the source of manganese-containing coordination complex includes a Mn(II)-containing component, e.g., "free" or uncoordinated Mn(II), in a weight ratio of Mn(II)-containing component to manganese-containing coordination complex of at least 1:100,000, e.g., at least 1:50,000, and even at least 1:15,000, and not more than 1:100, e.g., not more than 1:1,000, not more than 1:5,000, and / or not more than 1:8,000. For example, the weight ratio of Mn(II)-containing component to manganese-containing coordination complex in the source of manganese-containing coordination complex can be in the range of 1:100,000 to 1:100, and / or in the range of 1:75,000 to 1:1,000, and / or in the range of 1:50,000 to 1:5,000, and / or in the range of 1:15,000 to 1:8,000. As yet another example, when an aqueous formulation is prepared to provide a single dose of a manganese-containing coordination complex, such as for parenteral administration of a single dose of a pentaaza macrocycle complex, the Mn(II)-containing component (e.g., "free" Mn) can be at least 1 microgram, e.g., at least 10 micrograms, e.g., at least 50 micrograms, and even at least 100 micrograms, but typically less than about 2000 micrograms, e.g., less than 1000 micrograms, and even less than 850 micrograms of the Mn(II)-containing component.

[0063] In some embodiments, the chloride anion source comprises a salt capable of forming chloride anions in aqueous solution. For example, the chloride anion source can comprise at least one selected from the group consisting of sodium chloride, potassium chloride, calcium chloride, and magnesium chloride. In some embodiments, for example, when the aqueous formulation is intended for parenteral administration, the chloride anion source can be provided at a concentration and / or amount compatible with physiological conditions. For example, a chloride anion source (e.g., sodium chloride) can be added in an amount sufficient to provide a chloride anion concentration in the aqueous formulation of at least 100 mM, e.g., at least 110 mM, at least 115 mM, at least 120 mM, at least 130 mM, at least 145 mM, and / or at least 150 mM. For example, the chloride anion source can be provided in an amount sufficient to provide a chloride anion concentration in the aqueous formulation of 1000 mM or less, 200 mM or less, 180 mM or less, 175 mM or less, 160 mM or less, and / or 155 mM or less. For example, a chloride anion source (e.g., sodium chloride) may be provided in an amount to provide a chloride anion concentration in the aqueous formulation that is in the range of 100 mM to 200 mM, e.g., in the range of 130 mM to 160 mM and / or in the range of 145 mM to 158 mM, e.g., about 154 mM.

[0064] In some embodiments, the dianion source comprises at least one selected from the group consisting of a bicarbonate salt (e.g., sodium bicarbonate (NaHCO)) and a phosphate salt (e.g., sodium phosphate). In some embodiments, the dianion source comprises at least 0.1 mM, e.g., at least 0.25 mM, at least 1 mM, and / or at least 2.5 mM, and up to 26 mM of a dianion (e.g., CO). 2-) concentration, e.g., 15 mM or less, 10 mM or less, and / or 5 mM or less, e.g., in the range of 0.1 mM to 15 mM, and even in the range of 1 mM to 10 mM. By way of example, when a bicarbonate salt is provided as the dianion source, the concentration of the bicarbonate salt provided in the aqueous formulation can be at least 5 mM, e.g., at least 10 mM, at least 15 mM, at least 20 mM, and / or at least 25 mM, and 50 mM or less, e.g., 40 mM or less, 35 mM or less, and / or 30 mM or less. For example, the concentration of the bicarbonate salt provided in the aqueous formulation can be in the range of 5 mM to 50 mM, e.g., 15 mM to 40 mM, and even about 20 mM to 30 mM.

[0065] In yet another embodiment, a buffer system comprising one or more buffering agents may be provided in the aqueous formulation. According to certain embodiments herein, the dianion itself may be part of a buffering system, such as, for example, a bicarbonate buffering system and / or a phosphate buffering system, and may include a buffering agent that buffers the pH of the aqueous formulation in conjunction with its conjugate acid and / or conjugate base, which together form a buffering system. In certain embodiments, the buffering system comprises a buffering agent that acts as a dianion source and is provided at a concentration sufficient to buffer the aqueous formulation to a predetermined pH. In some embodiments, the buffering system is provided to buffer the aqueous formulation to a physiologically acceptable pH, for example, a pH within the range of 7 to 10, and even a pH within the range of 7.5 to 9. For example, in certain embodiments, the buffering agent may act as a dianion source and buffer the aqueous formulation to a predetermined pH, thereby providing a dianion concentration in the aqueous formulation that is consistent with buffering at that pH. In one example, when the buffering agent is sodium bicarbonate, an amount of sodium bicarbonate can be added to buffer the aqueous solution to a pH of about 8.3, with a concentration of sodium bicarbonate added to the formulation of about 26 mM.

[0066] Furthermore, according to certain embodiments, the amount of chloride anion source provided in the aqueous formulation is such that the concentration of chloride anions in the aqueous formulation exceeds the concentration of dianions in the aqueous formulation. That is, without being bound by any particular theory, the chloride anion source may be provided in an amount sufficient such that the concentration of chloride anions exceeds the dianions in the aqueous formulation, which is believed to, in certain instances, provide a protective effect to protect "free" Mn or other related Mn(II)-containing components from the dianions, thereby inhibiting and / or preventing the formation of precipitates that may otherwise form upon interaction with the dianions. In certain embodiments, the amount of chloride anion source and the amount of dianion source are provided in relative amounts such that the concentration of chloride anion in the aqueous formulation exceeds the concentration of dianion in the formulation by at least 10:1, at least 100:1, at least 250:1, at least 500:1, at least 750:1, at least 1000:1, at least 5000:1, and / or at least 10,000:1 in terms of mol / L concentration ratio of chloride anion to dianion in the aqueous formulation.

[0067] In some embodiments, the dianion source can be provided in an aqueous solution for combination with the manganese-containing coordination complex source simultaneously with the chloride anion source. For example, in some embodiments, a chloride anion source (e.g., sodium chloride) and a dianion source (e.g., bicarbonate) can be combined to form an aqueous solution. The aqueous solution can then be combined with the manganese-containing coordination complex source, for example, by combining the aqueous solution containing chloride anions and dianions with another aqueous solution of the manganese-containing coordination complex, or by otherwise adding the manganese-containing coordination complex to the aqueous solution containing chloride anions and dianions (e.g., by dissolving the manganese-containing coordination complex source in the aqueous solution containing chloride anions and dianions). For example, in some embodiments, an aqueous solution of the manganese-containing coordination complex is formed by dissolving the manganese-containing coordination complex source in water and optionally adjusting the pH. The separate aqueous solution is prepared by combining the chloride anion source and the dianion source, for example, in amounts sufficient to provide a desired concentration of chloride anions and dianions (e.g., excess chloride ion concentration) in the final aqueous formulation. An aqueous solution of chloride anions and dianions is then added to the aqueous solution of the manganese-containing coordination complex to form an aqueous formulation comprising the manganese-containing coordination complex, chloride anions, and dianions, with the chloride anions present at a concentration greater than the dianion concentration in the formulation. In yet another embodiment, an aqueous solution comprising a chloride anion source and a dianion source is prepared, for example, in an amount sufficient to provide a predetermined chloride anion and dianion concentration in the final aqueous formulation (e.g., having a chloride anion concentration greater than the dianion concentration in the final aqueous formulation). The manganese-containing coordination complex source can then be added directly to and / or dissolved in the aqueous solution to provide the final aqueous formulation.

[0068] According to yet another embodiment, the dianion source is added after combining the chloride anion source with the manganese-containing coordination complex source in aqueous solution. For example, an aqueous solution of a manganese-containing coordination complex can be formed by dissolving the manganese-containing coordination complex source in water and adjusting the pH. The chloride anion source can be added to the aqueous solution containing the manganese-containing coordination complex, for example, by adding the chloride anion source (e.g., a chloride-containing salt) directly to the aqueous solution and dissolving the chloride anion source therein, and / or by providing a separate aqueous solution having the chloride anion source dissolved therein and then combining the separate aqueous solution with the aqueous solution containing the manganese-containing coordination complex. As yet another example, the chloride anion source can be dissolved in an aqueous solution, and the manganese-containing coordination complex source can be added directly thereto, dissolving the manganese-containing coordination complex therein. Once the aqueous solution containing chloride anions and a manganese-containing coordination complex is formed, a dianion source can be added thereto, for example, by adding a separate aqueous solution containing the dianion source to the aqueous solution containing chloride anions and a manganese-containing coordination complex, or by adding the dianion source directly (e.g., in salt form) to the aqueous solution containing chloride anions and a manganese-containing coordination complex. As with the simultaneous addition described above, the amounts of chloride anion source and dianion source provided are such that the chloride anion concentration exceeds the dianion concentration in the final aqueous formulation.

[0069] According to embodiments described herein, the dianion source is added simultaneously with or after combining the chloride anion with the manganese-containing coordination complex. In some embodiments, substantially the entire amount of the dianion source is added simultaneously with or after combining the chloride anion with the manganese-containing coordination complex, such that the dianion would not combine with the manganese-containing coordination complex in the absence of chloride anion. For example, at least 75 mol%, at least 85 mol%, at least 90 mol%, at least 95 mol%, at least 98 mol%, at least 99 mol%, and / or the entire molar amount of the dianion source added to form the aqueous formulation is added simultaneously with or after combining the chloride anion with the manganese-containing coordination complex. According to some embodiments, no amount of dianion source is combined with the manganese-containing coordination complex unless an excess amount of chloride anion is already present in the aqueous solution containing the manganese-containing coordination complex or is added simultaneously. Furthermore, according to certain embodiments, the dianion source is provided after combining the chloride anion with the manganese-containing coordination complex in aqueous solution, and the dianion source (e.g., bicarbonate salt) can be added thereto at least 30 seconds, at least 1 minute, at least 5 minutes, at least 10 minutes, at least 30 minutes, and / or at least 1 hour after combining the manganese-containing coordination complex with the chloride anion source in aqueous solution.

[0070] In one embodiment of the manufacturing method according to the embodiments described herein, the pH of filtered deionized water suitable for injection purposes is adjusted to about 7.5 using NaOH. A manganese-containing coordination complex, such as the pentaazamacrocycle complex corresponding to GC4419 described herein or another pentaazamacrocycle complex, is added to water in an amount of 9 mg / mL to form an aqueous solution thereof. Sodium chloride salt is added to the aqueous solution to provide a 0.9% by weight solution. After the addition of the sodium chloride salt, sodium bicarbonate is added to the aqueous solution in an amount of 26 mM to buffer the solution. As further described in the Examples herein, the resulting aqueous formulation has good stability and shelf life, forming no visible precipitate after 9 months under appropriate storage conditions, including maintaining at a temperature of about 5-8°C.

[0071] According to certain embodiments, the aqueous formulations can be used in methods for treating conditions in patients. For example, the aqueous formulations can be used for parenteral administration of a buffered solution containing an aqueous formulation of a manganese-containing coordination complex. In some instances, the aqueous formulations can be used for intravenous administration of the manganese-containing coordination complex. Additional treatment methods using the aqueous formulations, as well as disease states and conditions that can be treated therewith, are described in more detail below.

[0072] In further embodiments, aspects of the present invention relate to buffered formulations, including aqueous formulations, e.g., buffered formulations of manganese-containing pentaaza macrocyclic complexes. The buffered formulations can include, for example, aqueous formulations prepared by any of the process embodiments described herein. In some embodiments, the buffered aqueous solution can contain (i) the pentaaza macrocyclic complex at a concentration of 2 mM to 100 mM, (ii) sodium chloride at a concentration of 130 mM to 160 mM, and a buffering agent comprising sodium bicarbonate at a concentration sufficient to buffer the aqueous solution to a pH range of 7 to 10, e.g., 20 mM to 30 mM. For example, in some embodiments, the buffered aqueous solution can contain the pentaaza macrocyclic complex at a concentration of at least 2 mM, at least 6 mM, at least 18 mM, at least 20 mM, and / or at least 40 mM, but less than 100 mM.

[0073] For example, in certain embodiments, the buffered aqueous solution can contain the pentaaza macrocycle complex at a concentration of at least 1 mg / mL, at least 3 mg / mL, at least 5 mg / mL, at least 9 mg / mL, at least 15 mg / mL, at least 18 mg / mL, and / or at least 20 mg / mL, but generally not more than 100 mg / mL, e.g., not more than 75 mg / mL, not more than 50 mg / mL, not more than 30 mg / mL, not more than 20 mg / mL, and / or not more than 10 mg / mL. For example, the concentration of the pentaaza macrocycle complex can be in the range of 1 mg / mL to 50 mg / mL, e.g., in the range of 5 mg / mL to 15 mg / mL, and even in the range of 3 mg / mL to 10 mg / mL. For example, the aqueous formulation may contain a pentaaza macrocycle complex (e.g., GC4419) in an amount of at least 10 mg, at least 25 mg, at least 30 mg, at least 50 mg, at least 65 mg, at least 70 mg, at least 75 mg, at least 80 mg, at least 85 mg, at least 90 mg, at least 95 mg, at least 100 mg, at least 105 mg, at least 110 mg, at least 115 mg, and / or at least 120 mg, but generally not more than 500 mg.

[0074] When prepared as described herein (e.g., by combining sodium bicarbonate simultaneously with or after combining sodium chloride with the pentaaza macrocycle complex), the buffered formulation can exhibit good storage stability. For example, in one embodiment, the storage stability of the buffered formulation is such that no manganese-containing precipitate is discernible by visual detection for 9 months after preparation of the buffered formulation. In yet another embodiment, the storage stability is such that no manganese-containing precipitate is discernible by visual detection 1 day and / or 6 days after formation of the buffered formulation. Visual detection can include visual inspection of the buffered solution to determine whether a manganese-containing precipitate has formed in solution. In further embodiments, the buffered formulation can be prepared by any of the methods described herein.

[0075] As yet another example, an ICP-MS (inductively coupled plasma-mass spectrometry) storage stability assay can be performed to determine the amount of manganese-containing precipitate that forms in a buffered formulation over time. According to some embodiments, the ICP-MS storage stability assay can include filtering the buffered formulation through a 0.45 micrometer filter, washing the residue with water at pH 8.0, dissolving the residue with nitric acid, and performing inductively coupled mass spectrometry (ICP-MS) to detect the manganese content of any precipitate. According to embodiments herein, the amount of manganese measured by the ICP-MS storage stability assay after at least 1 day, at least 6 days, and / or at least 9 months is less than 1500 ppm and / or even less than 1200 ppm.

[0076] Further details and / or embodiments of the aqueous formulation are provided below, including further description of the components of the aqueous formulation and optional additives thereto and methods of treatment therewith.

[0077] Manganese-containing pentaaza macrocyclic complexes In some embodiments, the pentaaza macrocycle complex corresponds to a complex of formula (I): [ka] [During the ceremony, M is Mn 2+ or Mn 3+ and; R1, R2, R'2, R3, R4, R5, R'5, R6, R'6, R7, R8, R9, R'9 and R 10 are independently hydrogen, hydrocarbyl, substituted hydrocarbyl, heterocyclyl, an amino acid side chain moiety, or -OR 11 , -NR 11 R 12 , -COR 11 , -CO2R 11 , -CONR 11 R 12 , -SR 11 , -SOR 11 , -SO2R 11 , -SO2NR11 R 12 , -N(OR 11 )(R 12 ), -P(O)(OR 11 )(OR 12 ), -P(O)(OR 11 )(R 12 ) and -OP(O)(OR 11 )(OR 12 ), wherein R 11 and R 12 are independently hydrogen or alkyl; U together with adjacent carbon atoms of the macrocyclic ring form a fused substituted or unsubstituted, saturated, partially saturated or unsaturated ring or heterocycle having 3 to 20 ring carbon atoms; V together with adjacent carbon atoms of the macrocyclic ring forms a fused substituted or unsubstituted, saturated, partially saturated or unsaturated ring or heterocycle having 3 to 20 ring carbon atoms; W, together with the nitrogen of the macrocyclic ring to which it is attached and the carbon atoms of the macrocyclic ring, forms an aromatic or alicyclic, substituted or unsubstituted, saturated, partially saturated, or unsaturated nitrogen-containing fused heterocycle having 2 to 20 ring carbon atoms, provided that when W is a fused aromatic heterocycle, the hydrogens attached to the nitrogen that are part of both the heterocyclic ring and the macrocyclic ring and the R and R attached to the carbon atoms that are part of both the heterocyclic ring and the macrocyclic ring are not included. 10 does not exist; X and Y represent suitable ligands or their corresponding anions from any monodentate or polydentate ligand or ligand system; Z is a counterion; n is an integer from 0 to 3; The dashed lines represent the coordinate bonds between the nitrogen atom of the macrocycle and the transition metal, manganese.

[0078] As described above for the pentaaza macrocycle complexes of formula (I), M is Mn 2+ or Mn 3+ In certain embodiments where the pentaaza macrocycle complex corresponds to Formula (I), M is Mn 2+In another particular embodiment where the pentaaza macrocycle complex corresponds to Formula (I), M is Mn 3+ is.

[0079] R1, R2, R'2, R3, R4, R5, R'5, R6, R'6, R7, R8, R9, R'9 and R 10 In embodiments where one or more of is hydrocarbyl, for example, suitable hydrocarbyl moieties include, but are not limited to, alkenyl, alkenylcycloalkenyl, alkenylcycloalkyl, alkyl, alkylcycloalkenyl, alkylcycloalkyl, alkynyl, aralkyl, aryl, cycloalkenyl, cycloalkyl, cycloalkylalkyl, cycloalkylcycloalkyl, cycloalkenylalkyl, and aralkyl. In some embodiments, R, R, R', R, R, R', R, R, and R 10 are independently hydrogen, hydrocarbyl, substituted hydrocarbyl, or heterocyclyl. More preferably, in this embodiment, R, R, R', R, R, R', R, R, R', and R 10 are independently hydrogen or lower alkyl (e.g., C-C alkyl, more typically C-C alkyl). Thus, for example, R, R, R', R, R, R, R', R, R, R', and R 10 may independently be hydrogen, methyl, ethyl, propyl, or butyl (linear, branched, or cyclic). In certain preferred embodiments, R, R, R', R, R, R, R', R, R, R', R, R, R, and R 10 are independently hydrogen or methyl.

[0080] In certain preferred embodiments where the pentaaza macrocycle complex corresponds to formula (I), R, R', R, R, R, R', R, R, R, R' and R 10are each hydrogen, one of R and R' is hydrogen, and the other of R and R' is methyl. In this embodiment, for example, R, R, R', R, R, R, R', R, R, R, R' and R 10 can each be hydrogen and R'6 is methyl. Alternatively, for example, R1, R2, R'2, R3, R4, R5, R'5, R'6, R7, R8, R9, R'9 and R 10 can each be hydrogen and R is methyl. In another preferred embodiment, wherein the pentaaza macrocycle complex corresponds to formula (I), R, R, R, R', R', R, R and R 10 are each hydrogen, one of R and R' is hydrogen, the other of R and R' is methyl, one of R and R' is hydrogen, and the other of R and R' is methyl. In this embodiment, for example, R, R', R, R, R, R', R, R, R, R, and R 10 can each be hydrogen, and R and R' are methyl. Alternatively, for example, R, R, R, R, R, R', R, R, R' and R 10 can each be hydrogen and R'2 and R9 are methyl. In another embodiment where the pentaaza macrocycle complex corresponds to Formula (I), R1, R2, R'2, R3, R4, R5, R'5, R6, R'6, R7, R8, R9, R'9 and R 10 are each hydrogen.

[0081] In certain embodiments, the U and V moieties are independently substituted or unsubstituted fused cycloalkyl moieties having from 3 to 20 ring carbon atoms, more preferably from 4 to 10 ring carbon atoms. In certain embodiments, the U and V moieties are each a trans-cyclohexanyl fused ring.

[0082] In certain embodiments, W moiety is a substituted or unsubstituted fused heteroaromatic moiety.In certain embodiments, W moiety is a substituted or unsubstituted fused pyridino moiety.When W is a substituted fused pyridino moiety, for example, W moiety is typically substituted with hydrocarbyl or substituted hydrocarbyl moiety (e.g., alkyl, substituted alkyl) at the ring carbon atom located at the para position of the nitrogen atom of heterocycle.In some preferred embodiments, W moiety is an unsubstituted fused pyridino moiety.

[0083] As above, X and Y represent suitable ligands or their corresponding anions (e.g., benzoic acid or benzoate anions, phenol or phenoxide anions, alcohol or alkoxide anions) from any monodentate or polydentate coordinated ligand or ligand system. For example, X and Y can be, among other possibilities, halo, oxo, aquo, hydroxo, alcohol, phenol, dioxygen, peroxo, hydroperoxo, alkylperoxo, arylperoxo, ammonia, alkylamino, arylamino, heterocycloalkylamino, heterocycloarylamino, amine oxide, hydrazine, alkylhydrazine, arylhydrazine, nitric oxide, cyanide, cyanate, thiocyanate, isocyanate, isothiocyanate, alkylnitrile, arylnitrile, alkylisonitrile, arylisonitrile, nitrate, nitrite, azide, alkylsulfonic acid, arylsulfonic acid, alkylsulfoxide, arylsulfoxide, alkylarylsulfoxide, alkylsulfenic acid, arylsulfenic acid, alkylsulfinic acid, arylsulfinic acid, alkylthiolcarboxylic acid, arylthiolcarboxylic acid, alkylthiolthiocarboxylic acid, aryl ... carboxylic acids, alkylcarboxylic acids, arylcarboxylic acids, urea, alkylureas, arylureas, alkylarylureas, thioureas, alkylthioureas, arylthioureas, alkylarylthioureas, sulfates, sulfites, bisulfates, bisulfites, thiosulfates, thiosulfites, hydrosulfites, alkylphosphines, arylphosphines, alkylphosphine oxides, arylphosphine oxides, alkylarylphosphine oxides, alkylphosphine sulfides, arylphosphine sulfides, alkylarylphosphine sulfides, alkylphosphonic acids, arylphosphonic acids, alkylphosphinic acids, arylphosphinic acids, alkylphosphinous acids, arylphosphinous acids, phosphates, thiophosphates, phosphites, pyrophosphites, triphosphates, hydrogen phosphates, dihydrogen phosphates, alkylguanidino acids, arylguanidino acids, alkylarylguanidino acids, alkyl carbamates,The anion may be selected from the group consisting of aryl carbamates, alkylaryl carbamates, alkylthiocarbamates, arylthiocarbamates, alkylarylthiocarbamates, alkyldithiocarbamates, aryldithiocarbamates, alkylaryldithiocarbamates, bicarbonates, carbonates, perchlorates, chlorates, chlorites, hypochlorites, perbromates, bromates, bromites, hypobromites, tetrahalomanganates, tetrafluoroborate, hexafluoroantimonate, hypophosphites, iodates, periodates, metaborate, tetraarylborate, tetraalkylborate, tartrates, salicylates, succinates, citrates, ascorbates, saccharinates, amino acids, hydroxamic acids, thiotosylates, and anions of ion exchange resins or their corresponding anions. In some embodiments, if present, X and Y are independently selected from the group consisting of halo, nitrate, and bicarbonate ligands. For example, in this embodiment, if present, X and Y are halo ligands, e.g., chloro ligands.

[0084] Further, in some embodiments, X and Y correspond to -OC(O)-X, where each X is -C(X)(X)(X), and each X is independently substituted or unsubstituted phenyl or -C(-X)(-X)(-X); each X2 is independently substituted or unsubstituted phenyl, methyl, ethyl, or propyl; Each X3 independently represents hydrogen, hydroxyl, methyl, ethyl, propyl, amino, -X5C(=O)R 13 wherein X5 is NH or O, and R 13 is C1-C 18 Alkyl, substituted or unsubstituted aryl or C1-C 18 aralkyl or -OR 14 where R 14 is C1-C 18 Alkyl, substituted or unsubstituted aryl or C1-C 18is aralkyl or together with X4 (=O); Each X4 is independently hydrogen, or together with X3 is (=O).

[0085] In yet another embodiment, X and Y are independently selected from the group consisting of charge-neutralizing anions derived from any monodentate and polydentate ligands and ligand systems and their corresponding anions; or X and Y are independently selected from the group consisting of R, R, R', R, R, R, R', R, R, R', R, R, R, and R 10 binds to one or more of

[0086] In the pentaaza macrocycle complexes corresponding to formula (I), Z is a counterion (e.g., a charge-neutralizing anion) and n is an integer from 0 to 3. In general, Z can correspond to the counterions of the moieties recited for X and Y.

[0087] In combination, among certain preferred embodiments, there are pentaaza macrocyclic complexes corresponding to formula (I): M is Mn 2+ or Mn 3+ and; R1, R2, R'2, R3, R4, R5, R'5, R6, R'6, R7, R8, R9, R'9 and R 10 are independently hydrogen or lower alkyl; U and V are each a trans-cyclohexanyl fused ring; W is a substituted or unsubstituted fused pyridino moiety; X and Y are ligands; Z, if present, is a charge-neutralizing anion; Pentaaza macrocyclic complexes exist.

[0088] More preferably, in these embodiments, M is Mn 2+ and R1, R2, R'2, R3, R4, R5, R'5, R6, R'6, R7, R8, R9, R'9 and R 10are independently hydrogen or methyl; U and V are each a trans-cyclohexanyl fused ring; W is an unsubstituted fused pyridino moiety; X and Y are independently a halo ligand (e.g., fluoro, chloro, bromo, iodo). Z, if present, can be a halide anion (e.g., fluoride, chloride, bromide, iodide).

[0089] In yet another embodiment, the pentaaza macrocycle complex has the following formula (II): [ka] [During the ceremony, X and Y represent suitable ligands or their corresponding anions from any monodentate or polydentate ligand or ligand system; R A , R B , R C and R D are independently hydrogen, hydrocarbyl, substituted hydrocarbyl, heterocyclyl, an amino acid side chain moiety, or -OR 11 , -NR 11 R 12 , -COR 11 , -CO2R 11 , -CONR 11 R 12 , -SR 11 , -SOR 11 , -SO2R 11 , -SO2NR 11 R 12 , -N(OR 11 )(R 12 ), -P(O)(OR 11 )(OR 12 ), -P(O)(OR 11 )(R 12 ) and -OP(O)(OR 11 )(OR 12 ), wherein R 11 and R 12 are independently hydrogen or alkyl. It is expressed by:

[0090] Further, in some embodiments, the pentaaza macrocycle complex has formula (III) or formula (IV): [ka] [During the ceremony, X and Y represent suitable ligands or their corresponding anions from any monodentate or polydentate ligand or ligand system; R A , R B , R C and R D are independently hydrogen, hydrocarbyl, substituted hydrocarbyl, heterocyclyl, an amino acid side chain moiety, or -OR 11 , -NR 11 R 12 , -COR 11 , -CO2R 11 , -CONR 11 R 12 , -SR 11 , -SOR 11 , -SO2R 11 , -SO2NR 11 R 12 , -N(OR 11 )(R 12 ), -P(O)(OR 11 )(OR 12 ), -P(O)(OR 11 )(R 12 ) and -OP(O)(OR 11 )(OR 12 ), wherein R 11 and R 12 are independently hydrogen or alkyl. It is expressed by:

[0091] In yet another embodiment, the pentaaza macrocycle complex is a compound represented by a formula selected from the group consisting of Formulas (V)-(XVI): [ka] [ka] [ka]

[0092] In some embodiments, X and Y in any formula described herein are independently selected from the group consisting of fluoro, chloro, bromo, and iodo anion. In yet other embodiments, X and Y in any formula described herein are independently selected from the group consisting of alkyl carboxylate, aryl carboxylate, and aryl alkyl carboxylate. In yet other embodiments, X and Y in any formula described herein are independently an amino acid.

[0093] In some embodiments, the pentaaza macrocyclic ring complex has the following formula (IA): [ka] [During the ceremony, M is Mn 2+ or Mn 3+ and; R 1A , R 1B , R2, R3, R 4A , R 4B , R5, R6, R 7A , R 7B , R8, R9, R 10A , and R 10B are independently hydrogen, hydrocarbyl, substituted hydrocarbyl, heterocyclyl, an amino acid side chain moiety, or -OR 11 , -NR 11 R 12 , -COR 11 , -CO2R 11 , -C(=O)NR 11 R 12 , -SR 11 , -SOR 11 , -SO2R 11 , -SO2NR 11 R 12 , -N(OR 11 )(R 12 ), -P(=O)(OR 11 )(OR12 ), -P(=O)(OR 11 )(R 12 ) and -OP(=O)(OR 11 )(OR 12 ), wherein said R 11 and R 12 are independently hydrogen or alkyl; U together with adjacent carbon atoms of the macrocyclic ring form a fused substituted or unsubstituted, saturated, partially saturated or unsaturated ring or heterocycle having 3 to 20 ring carbon atoms; V together with adjacent carbon atoms of the macrocyclic ring forms a fused substituted or unsubstituted, saturated, partially saturated or unsaturated ring or heterocycle having 3 to 20 ring carbon atoms; W, together with the nitrogen of the macrocyclic ring to which it is attached and the carbon atoms of the macrocyclic ring, forms an aromatic or alicyclic, substituted or unsubstituted, saturated, partially saturated or unsaturated, nitrogen-containing fused heterocycle having 2 to 20 ring carbon atoms, provided that when W is a fused aromatic heterocycle, the hydrogens attached to the nitrogen that are part of both the heterocyclic ring and the macrocyclic ring and the R and R attached to the carbon atoms that are part of both the heterocyclic ring and the macrocyclic ring are not substituted. 10 does not exist; where each X1 is independently substituted or unsubstituted phenyl or -C(-X2)(-X3)(-X4); each X2 is independently substituted or unsubstituted phenyl or alkyl; Each X3 independently represents hydrogen, hydroxyl, alkyl, amino, or -X5C(=O)R 13 wherein X5 is NH or O, and R 13 is C1-C 18 Alkyl, substituted or unsubstituted aryl or C1-C 18 Aralkyl or -OR 14 where R 14 is C1-C 18 Alkyl, substituted or unsubstituted aryl or C1-C 18 aralkyl, or together with X4 (=O); each X4 independently is hydrogen or together with X3 is (=O); The bond between the transition metal M and the macrocyclic nitrogen atom and the bond between the transition metal M and the oxygen atom of the axial ligand -OC(=O)X1 are coordinate covalent bonds. It has.

[0094] In some embodiments, within Formula (IA) and groups contained therein, in one group of compounds, X1 is -C(-X2)(-X3)(-X4), and each combination of X2, X3, and X4 corresponds to one of the combinations shown in the following table: [Table 1]

[0095] Furthermore, within formula (IA) and the groups contained therein, in one group of compounds, X1 is C(-X2)(-X3)(-X4) and X3 is -X5C(=O)R 13 and the combination of X2, X3 and X4 includes any of the combinations shown in the following table: [Table 2] where R 13 is C1-C 18 Alkyl, substituted or unsubstituted aryl or C1-C 18 Aralkyl or -OR 14 where R 14 is C1-C 18 Alkyl, substituted or unsubstituted aryl or C1-C 18 It is aralkyl.

[0096] In some embodiments, the pentaaza macrocyclic complex corresponding to formula (IA) is one of the complex formulas (IE), e.g., formula (IE R1 ), expression (IE S1 ), expression (IE R2 ), expression (IE S2 ), expression (IE R3 ) or expression (IES3 ) corresponds to [ka] [During the ceremony, M is Mn 2+ or Mn 3+ and; each X1 is independently substituted or unsubstituted phenyl or -C(X2)(X3)(X4); each X2 is independently substituted or unsubstituted phenyl, methyl, ethyl, or propyl; each X3 independently is hydrogen, hydroxyl, methyl, ethyl, propyl, amino, or together with X4 is =O; each X4 independently is hydrogen or together with X3 is =O; The bond between manganese and the macrocyclic nitrogen atom and the bond between manganese and the oxygen atom of the axial ligand, -OC(O)X1, are coordinate covalent bonds.

[0097] In some embodiments, each X1 is -C(X2)(X3)(X4), where each -C(X2)(X3)(X4) corresponds to any of combinations 1-9 found in the table for formula (IA) above.

[0098] In yet another embodiment, X and Y in the pentaaza macrocycle complex of Formula (I) correspond to the ligands in Formula (IA) or Formula (IE). For example, X and Y in the complex of Formula (I) can correspond to -OC(O)-X, where X is as defined above for the complexes of Formula (IA) and Formula (IE).

[0099] In certain embodiments, the pentaaza macrocyclic ring complex corresponding to Formula (I) (e.g., Formula (I) or any of the subgroups of Formula (I) corresponding to Formulas (II)-(XIV), Formula (IA) and Formula (IE)) may comprise any of the following structures: [ka] [ka] [ka] [ka] [ka]

[0100] In certain embodiments, pentaaza macrocyclic ring complexes for use in the methods and compositions described herein include pentaaza macrocyclic ring complexes corresponding to Formula (2), Formula (3), Formula (4), Formula (5), Formula (6), and Formula (7). [ka] wherein X and Y in each of formulas (2), (3), (4), (5), (6), and (7) are independently a ligand. For example, according to certain embodiments, the pentaaza macrocycle complexes described herein and for use in the compositions include pentaaza macrocycle complexes corresponding to formulas (2), (3), (4), (5), (6), and (7), wherein X and Y in each of these formulas are halo, e.g., chloro. Alternatively, X and Y are ligands other than chloro, e.g., any of the ligands described above.

[0101] In another embodiment, the pentaaza macrocycle complex corresponds to formula (6) or formula (7). [ka]

[0102] The chemical structures of 6 (e.g., the dichloro complex form described in Riley, DP, Schall, OF, 2007, Advances in Inorganic Chemistry, 59: 233-263) and 7 (e.g., the dichloro complex form of 7) described herein are identical except that they possess mirror image chirality; i.e., the enantiomeric structures are not superimposable.

[0103] For example, the pentaaza macrocycle complex may correspond to at least one of the following complexes: [ka]

[0104] In yet another embodiment, the pentaaza macrocycle complex may correspond to at least one of the following complexes and / or enantiomers thereof: [ka]

[0105] In certain embodiments, the enantiomeric purity of the pentaaza macrocyclic ring complex is greater than 95%, more preferably greater than 98%, more preferably greater than 99%, and most preferably greater than 99.5%. As used herein, the term "enantiomeric purity" refers to the amount of a compound having a designated absolute stereochemistry expressed as a percentage of the designated compound and its total amount. In certain embodiments, the diastereomeric purity of the pentaaza macrocyclic ring complex is greater than 98%, more preferably greater than 99%, and most preferably greater than 99.5%. As used herein, the term "diastereomeric purity" refers to the amount of a compound having a designated absolute stereochemistry expressed as a percentage of the designated compound and its total amount of diastereomers. Methods for determining diastereomeric and enantiomeric purity are known in the art. Diastereomeric purity can be determined by any analytical method capable of quantitatively distinguishing between a compound and its diastereomers, such as high-performance liquid chromatography (HPLC). Similarly, enantiomeric purity can be determined by any analytical method capable of quantitatively distinguishing between a compound and its enantiomers. Examples of suitable analytical methods for determining proper enantiomeric purity include, but are not limited to, optical rotation of plane polarized light using a polarimeter and HPLC using chiral column packing materials.

[0106] In certain embodiments, a therapeutically effective amount of a pentaaza macrocyclic ring complex can be an amount sufficient to provide a peak plasma concentration of at least 0.1 μM when administered to a patient. For example, in certain embodiments, a pentaaza macrocyclic ring complex can be administered in an amount sufficient to provide a peak plasma concentration of at least 0.1 μM when administered to a patient. In yet other embodiments, a pentaaza macrocyclic ring complex can be administered in an amount sufficient to provide a peak plasma concentration of at least 10 μM when administered to a patient. Generally, a pentaaza macrocyclic ring complex is not administered in an amount that would provide a peak plasma concentration of greater than 40 μM when administered to a patient. For example, a pentaaza macrocyclic ring complex can be administered in an amount sufficient to provide a peak plasma concentration in the range of 0.1 μM to 40 μM in a patient. As another example, a pentaaza macrocyclic ring complex can be administered in an amount sufficient to provide a peak plasma concentration in the range of 0.5 μM to 20 μM in a patient. As another example, the pentaaza macrocyclic complex may be administered in an amount sufficient to provide a peak plasma concentration in the range of 1 μM to 10 μM in a patient.

[0107] In yet another embodiment, the dose of the pentaazamacrocycle complex administered per kg of patient body weight can be at least 0.1 mg / kg, e.g., at least 0.2 mg / kg. For example, the dose of the pentaazamacrocycle complex administered per kg of patient body weight can be at least 0.5 mg / kg. As another example, the dose of the pentaazamacrocycle complex administered per kg of patient body weight can be at least 1 mg / kg. In another example, the dose of the pentaazamacrocycle administered per kg of patient body weight can be at least 2 mg / kg, e.g., at least 3 mg / kg, and even at least about 15 mg / kg, e.g., at least 24 mg / kg, and even at least 40 mg / kg. Generally, the dose of the pentaazamacrocycle administered per kg of patient body weight will not exceed 1000 mg / kg. For example, the dose of the pentaazamacrocycle compound administered per kg of patient body weight can be in the range of 0.1 to 1000 mg / kg, e.g., 0.2 mg / kg to 40 mg / kg, e.g., 0.2 mg / kg to 24 mg / kg, and even 0.2 mg / kg to 10 mg / kg. As another example, the dose of the pentaazamacrocycle complex administered per kg of patient body weight can be in the range of 1 mg / kg to 1000 mg / kg, e.g., 3 mg / kg to 1000 mg / kg, and even 5 mg / kg to 1000 mg / kg, e.g., 10 mg / kg to 1000 mg / kg. As another example, the dose of the pentaazamacrocycle complex administered per kg of patient body weight can be in the range of 2 mg / kg to 15 mg / kg. As another example, the dose of the pentaazamacrocycle complex administered per kg of patient body weight can be in the range of 3 mg / kg to 10 mg / kg. As another example, the dose of the pentaazamacrocycle administered per kg of patient body weight can be in the range of 0.5 to 5 mg / kg. As a further example, the dose of the pentaazamacrocycle administered per kg of patient body weight can be in the range of 1 to 5 mg / kg.

[0108] In certain embodiments, the dose of the pentaaza macrocycle complex can be at least 15 mg, at least 30 mg, at least 50 mg, at least 75 mg, at least 90 mg, at least 100 mg, and / or at least 112 mg. The dose of the pentaaza macrocycle complex can also be administered over a defined infusion period, such as at a rate corresponding to a 15-minute, 30-minute, 45-minute, 60-minute infusion period, and / or longer infusion period. According to certain embodiments, a pentaaza macrocycle complex, such as GC4419, can be administered over a 1-hour period at an infusion rate corresponding to at least 75 mg and / or at least 90 mg.

[0109] In certain embodiments, the above dosages and / or plasma concentrations are particularly appropriate for pentaaza macrocyclic complexes corresponding to GC4419, but they are also appropriate for other pentaaza macrocyclic complexes. Furthermore, one skilled in the art will recognize how to adjust the dosage and / or plasma concentration based on factors such as the molecular weight and / or activity of the particular compound used. For example, for pentaaza macrocyclic complexes with twice the activity of GC4419, the dosage and / or plasma concentration can be halved, or for pentaaza macrocyclic complexes with molecular weights greater than that of GC4419, correspondingly higher dosages can be used.

[0110] The administration schedule of the pentaaza macrocyclic complex can also be selected depending on the intended treatment. For example, in some embodiments, a suitable administration schedule can include administering to the patient at least one day per week, e.g., at least two, three, four, five, six, or seven days per week (e.g., daily) during treatment. As another example, in some embodiments, administration can be at least once per day (qd), or even at least twice per day (bid).

[0111] Treatment method While the treatment of conditions, including oral mucositis, cancer, or other conditions described herein, includes achieving a therapeutic benefit, therapeutic agents can also be administered to achieve a preventative benefit. Therapeutic benefit generally refers to at least partial eradication or amelioration of the existing disorder being treated. For example, in a cancer patient, therapeutic benefit includes the (partial or complete) disappearance of the existing cancer. Therapeutic benefit can also be achieved with at least partial or complete eradication or amelioration of one or more physiological symptoms associated with the existing disorder, such that improvement is observed in the patient despite the fact that the patient may still suffer from the existing disorder. For preventative benefit, the methods of the present invention can be performed on cancer patients, or the compositions of the present invention can be administered to patients at risk of progression or who report one or more physiological symptoms of such a condition, even if a diagnosis of the condition has not been made.

[0112] Generally, any subject who has or is thought to have a condition or disorder can be treated using the compositions and methods of the present invention. The subject who is treated by the methods described herein is a mammalian subject, and typically a human patient. Other mammals that can be treated by the present invention include companion animals such as dogs and cats, livestock such as cows, horses and pigs, as well as birds and more exotic animals (e.g., animals found in zoos or nature reserves).

[0113] According to one aspect of the present invention, described herein are methods for treating tissue damage resulting from cancer treatment (e.g., radiation therapy or chemotherapy) delivered to a subject in need of treatment. According to another aspect of the present invention, described herein are methods for treating a human patient for tissue damage resulting from exposure to radiation. Thus, in various embodiments, for example, exposure to radiation can be accidental, unintentional, or intentional. As noted above, the treatment of tissue damage described herein can include both inhibition (i.e., prevention) and amelioration of any tissue damage resulting from development or activity. Generally, the method comprises administering a therapeutically effective amount of a pentaaza macrocycle complex to a subject. In a preferred embodiment, the complex is a dichloro complex of formula (GC4419), although other pentaaza macrocycle complexes described herein can also be used.

[0114] Treatment of tissue damage resulting from cancer treatment or other radiation exposure using the methods described herein involves administering a therapeutically effective amount of a pentaaza macrocyclic complex, such as, but not limited to, GC4419. Generally, a range of therapeutically effective amounts can be used, depending, for example, on the compound selected and its safety and efficacy, the type, location, and severity of the tissue damage, among other factors. Examples of tissue damage that can be treated include oral mucositis and other forms of tissue damage, including tissue damage affecting the mucosal lining of the upper and lower digestive tract.

[0115] In another embodiment, the preparation can be used for the treatment of cancer and / or tumor.Cancer and tumor generally refer to and describe physiological conditions in mammals that are typically characterized by uncontrolled cell proliferation.The pharmaceutical preparations described herein can treat a variety of tumors, such as tumors of the breast, heart, lung, small intestine, colon, spleen, kidney, bladder, head and neck, ovary, prostate, brain, pancreas, skin, bone, bone marrow, blood, thymus, uterus, testicle, cervix and liver.

[0116] In certain embodiments, the tumor or cancer is selected from adenoma, angiosarcoma, astrocytoma, epithelial carcinoma, germinoma, glioblastoma, glioma, hamartoma, hemangioendothelioma, angiosarcoma, hematoma, hepatoblastoma, leukemia, lymphoma, medulloblastoma, melanoma, neuroblastoma, osteosarcoma, retinoblastoma, rhabdomyosarcoma, sarcoma, and teratoma. Tumors include acral lentiginous melanoma, actinic keratosis, adenocarcinoma, adenoid cystic carcinoma, adenoma, adenosarcoma, adenosquamous carcinoma, astrocytic tumor, Bartholin's gland carcinoma, basal cell carcinoma, bronchial adenocarcinoma, capillary carcinoma, carcinoid, carcinoma, carcinosarcoma, cavernous carcinoma, cholangiocarcinoma, chondrosarcoma, choroid plexus papilloma / carcinoma, clear cell, cystadenoma, yolk sac tumor, endometrial hyperplasia, endometrial stromal sarcoma, endometrioid adenocarcinoma, ependymal tumor, epithelioid cell carcinoma, Ewing's sarcoma, fibrolamellar carcinoma, focal nodular hyperplasia, gastrinoma, germ cell tumor, glioblastoma, glucagonoma, hemangioblastoma, hemangioendothelioma, hemangioma, liver adenoma, liver adenomatosis, hepatocellular carcinoma, insulinoma, intraepithelial neoplastic lesion, and intraepithelial squamous cell carcinoma. The tumor may be selected from: invasive squamous cell carcinoma, large cell carcinoma, leiomyosarcoma, lentigo maligna melanoma, malignant melanoma, malignant mesothelioma, medulloblastoma, medulloepithelioma, melanoma, meningioma, mesothelioma, metastatic carcinoma, mucoepidermoid carcinoma, neuroblastoma, neuroepithelial adenocarcinoma, nodular melanoma, oat cell carcinoma, oligodendroglioma, osteosarcoma, pancreatic carcinoma, papillary serous adenocarcinoma, pineal cell carcinoma, pituitary tumor, plasmacytoma, pseudosarcoma, pulmonary blastoma, renal cell carcinoma, retinoblastoma, rhabdomyosarcoma, sarcoma, serous carcinoma, small cell carcinoma, soft tissue carcinoma, somatostatinoma, squamous cell carcinoma, squamous cell carcinoma, submesothelial carcinoma, superficial spreading melanoma, undifferentiated carcinoma, uveal melanoma, verrucous carcinoma, VIPoma, well-differentiated carcinoma, and Wilms' tumor.

[0117] Thus, for example, the present invention is directed to the treatment of, but not limited to, cancers of the bladder (including accelerated metastatic bladder cancer), breast, rectum (including colorectal cancer), kidney, liver, lung (including small cell and non-small cell lung cancer and lung adenocarcinoma), ovary, prostate, testis, genitourinary tract, lymphatic system, rectum, larynx, pancreas (including exocrine pancreatic cancer), esophagus, stomach, gallbladder, cervix, thyroid gland, and skin (including squamous cell carcinoma); leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma, and thyroid cancer. and other tumors including melanoma, xeroderma pigmentosum, keratoacanthoma, seminoma, follicular thyroid carcinoma, and teratocarcinoma.

[0118] Specific leukemias that may be treated using the formulations and methods described herein include, but are not limited to, acute nonlymphocytic leukemia, chronic lymphocytic leukemia, acute granulocytic leukemia, chronic granulocytic leukemia, acute promyelocytic leukemia, adult T-cell leukemia, aleukemic leukemia, leukemic leukemia, basophilic leukemia, blastic cell leukemia, bovine leukemia, chronic myeloid leukemia, leukemia cutis, fetal leukemia, eosinophilic leukemia, Gross' leukemia, hairy cell leukemia, hemoblastic leukemia, hemoblastic leukemia, histiocytic leukemia, stem cell leukemia, and leukemia. These include cell leukemia, acute monocytic leukemia, leukopenic leukemia, lymphocytic leukemia, lymphoblastic leukemia, lymphocytic leukemia, lymphoid leukemia, lymphosarcoma cell leukemia, mast cell leukemia, megakaryocytic leukemia, micromyeloblastic leukemia, monocytic leukemia, myeloblastic leukemia, myelocytic leukemia, myelogranulocytic leukemia, myelomonocytic leukemia, Naegeli leukemia, plasma cell leukemia, plasma cell leukemia, promyelocytic leukemia, Leader cell leukemia, Schilling leukemia, stem cell leukemia, subleukemic leukemia and undifferentiated cell leukemia.

[0119] Lymphomas can also be treated using the formulations and methods described herein. Lymphomas are generally neoplastic transformations of cells primarily present in lymphoid tissues. Lymphomas are tumors of the immune system and generally exist as both T-cell-related and B-cell-related diseases. There are two main distinct groups of lymphomas: non-Hodgkin's lymphoma (NHL) and Hodgkin's disease. These may include bone marrow, lymph nodes, spleen, and circulating cells. Treatment protocols involve removing bone marrow from patients, often using antibodies against antigens present in tumor cell types, and then preserving tumor cells. The patient is then given toxic doses of radiation or chemotherapy, and the purged bone marrow is then reinfused to repopulate the patient's hematopoietic system.

[0120] Other hematological tumors that can be treated using the combinations and methods described herein include myelodysplastic syndrome (MDS), myeloproliferative syndrome (MPS) and myeloma, such as solitary myeloma and multiple myeloma.Multiple myeloma (also called plasma cell myeloma) involves the skeletal system and is characterized by multiple masses of neoplastic plasma cells scattered throughout the system.It can also spread to lymph nodes and other parts, such as the skin.Solitary myeloma includes solitary lesions that tend to occur in the same location as multiple myeloma.

[0121] In some embodiments, the methods and formulations described herein are used to treat any of the following cancers: breast cancer, melanoma, oral squamous cell carcinoma, lung cancer including non-small cell lung cancer, renal cell carcinoma, colorectal cancer, prostate cancer, brain cancer, spindle cell carcinoma, urothelial carcinoma, bladder cancer, colorectal cancer, head and neck cancer such as squamous cell carcinoma, and pancreatic cancer. In yet another embodiment, the methods and formulations described herein are used to treat any of the following cancers: head and neck cancer and lung cancer.

[0122] As noted above, the disease or condition treated by the methods described herein can be any disease or condition treatable using a pentaaza macrocyclic complex. In some embodiments, for example, the disease or condition is selected from cancer, cardiovascular disorders, cerebrovascular disorders, skin disorders, fibrotic disorders, gastrointestinal disorders, immunological disorders, inflammatory disorders, metabolic disorders, neurological disorders, ophthalmic disorders, pulmonary disorders, infectious diseases, and combinations thereof. By way of example, uses include the treatment of inflammatory and hyperproliferative skin diseases and cutaneous manifestations of immunologically mediated diseases, such as psoriasis, atopic dermatitis, contact dermatitis and further eczematous dermatitis, seborrheic dermatitis, lichen planus, pemphigus, pemphigoid, epidermolysis bullosa, urticaria, angioedema, vasculitis, erythema, cutaneous eosinophilia, lupus erythematosus, comedones and alopecia areata; various ocular diseases (autoimmune and otherwise) such as keratoconjunctivitis, vernal conjunctivitis, uveitis associated with Behcet's disease, keratitis, herpetic keratitis, keratoconus, corneal epithelial dystrophy, corneal leukoplakia and ocular pemphigus. Additionally, reversible obstructive airway diseases, including asthma (e.g., bronchial asthma, allergic asthma, intrinsic asthma, extrinsic asthma, and dust asthma), particularly chronic or refractory asthma (e.g., late-onset asthma and airway hyperresponsiveness), bronchitis, allergic rhinitis, and the like, may be treated, prevented, and / or ameliorated by the methods described herein. Other treatable diseases and conditions include mucosal and vascular inflammation, such as gastric ulcers, vascular injury caused by ischemic disease, and thrombosis. Additionally, hyperproliferative vascular diseases, such as intimal smooth muscle cell hyperplasia, restenosis, and vascular obstruction, particularly following biologically or mechanically mediated vascular injury, may be treated by the compounds described herein.

[0123] Still other treatable diseases and conditions include, but are not limited to, cardiac diseases such as post-operative myocardial infarction, pulmonary diseases such as pulmonary muscle changes or remodeling and chronic obstructive pulmonary disease (COPD); intestinal inflammation / allergies such as ischemic bowel disease, inflammatory bowel disease, necrotizing enterocolitis, celiac disease, proctitis, eosinophilic gastroenteritis, mastocytosis, Crohn's disease, and ulcerative colitis; polymyositis, Guillain-Barré syndrome, Meniere's disease, polyneuritis, multiple myocardial infarction, and ulcerative colitis. neurological disorders such as septic shock and associated refractory hypotension; endocrine disorders such as hyperthyroidism and Graves' disease; arthritis (e.g., rheumatoid arthritis, chronic progressive arthritis, and osteoarthritis) and rheumatic diseases; pure red cell aplasia, aplastic anemia, aplastic anemia, idiopathic thrombocytopenic purpura, autoimmune hemolytic anemia, agranulocytosis, pernicious anemia, megaloblastoma blood disorders such as chronic anemia and red blood cell hypoplasia; bone diseases such as osteoporosis; respiratory diseases such as sarcoidosis, fibroid lung, and idiopathic interstitial pneumonia; skin diseases such as dermatomyositis, vitiligo, ichthyosis vulgaris, photoallergic sensitivity, and cutaneous T-cell lymphoma; cardiovascular diseases such as arteriosclerosis, atherosclerosis, aortitis syndrome, polyarteritis nodosa, and myocarditis; scleroderma, Wegener's granulomatosis, and Sjögren's disease collagen diseases such as leukemia, adiposity, eosinophilic fasciitis, periodontal diseases such as lesions of the gums, periodontium, alveolar bone and dental cementum, nephrotic syndromes such as glomerulonephritis, male pattern baldness or age-related alopecia by preventing hair loss or by providing hair growth and / or promoting hair growth and development, pyoderma and Sézary syndrome, Addison's disease, reactive oxygen-mediated diseases such as ischemia-reperfusion injury of organs (e.g., heart, liver, kidneys and gastrointestinal tract) resulting from protection, transplantation, organ failure (single or multiple) or ischemic diseases (e.g., thrombosis and myocardial infarction), movement disorders such as Parkinson's disease, nerve blockade-induced parkinsonism and tardive dyskinesia, intestinal diseases such as endotoxic shock, pseudomembranous colitis and colitis induced by drugs or radiation, renal diseases such as ischemic acute renal failure and chronic renal failure,These include pulmonary diseases such as intoxication caused by pulmonary oxygen or drugs (e.g., paraquat and bleomycin), lung cancer, and emphysema; eye diseases such as siderosis, retinitis, prurigo pigmentosa, senile macular degeneration, vitreous scarring, and corneal alkali burns; dermatitis such as erythema multiforme, linear IgA bullous dermatitis, and cementum dermatitis; and other diseases such as gingivitis, periodontitis, sepsis, pancreatitis, diseases caused by environmental pollution (e.g., air pollution), aging, carcinogenesis, cancer metastasis, and liver damage; diseases caused by histamine or leukotriene-C4 release; and Behcet's disease, such as intestinal, vascular, or neuro-Behcet's disease, and Behcet's disease affecting the mouth, skin, eyes, vulva, joints, epididymis, lungs, kidneys, etc. Furthermore, the compounds of the invention are useful for the treatment and prevention of liver diseases, such as immunogenic diseases (e.g., chronic autoimmune liver diseases such as autoimmune hepatitis, primary biliary cirrhosis and sclerosing cholangitis), partial hepatectomy, acute liver necrosis (e.g., necrosis caused by toxins, viral hepatitis, shock or anoxia), B viral hepatitis, non-A / non-B hepatitis, cirrhosis (e.g., alcoholic cirrhosis), and liver failure, such as fulminant liver failure, delayed liver failure and "acute exacerbation" liver failure (liver failure in the acute exacerbation of chronic liver disease), and They are useful for the treatment of bacterial or viral infections, particularly influenza or HIV infection, and further useful activities such as enhancing the effectiveness of chemotherapy, cytomegalovirus infection, particularly HCMV infection, anti-inflammatory activity, sclerosing and fibrotic diseases, such as nephrosis, scleroderma, fibrosis (e.g., idiopathic interstitial pneumonia, idiopathic pulmonary fibrosis, idiopathic mediastinal fibrosis, fibrosis complicating anti-tumor therapy, pulmonary fibrosis and lung fibrosis, including idiopathic interstitial pneumonia, idiopathic pulmonary fibrosis, idiopathic mediastinal fibrosis, fibrosis complicating anti-tumor therapy, radiation therapy, and chronic infections, including tuberculosis and aspergillosis and other fungal infections), arteriosclerosis, congestive heart failure, ventricular hypertrophy, post-operative adhesions and scarring, stroke, myocardial infarction, and injuries associated with ischemia and reperfusion;

[0124] Pharmaceutical preparations In certain embodiments, the aqueous solution may be administered by a parenteral route (e.g., intravenous, intraarterial, subcutaneous, rectal, subcutaneous, intramuscular, intraorbital, intravesical, intrathecal, intraperitoneal, or intrasternal). However, other routes of administration may also be possible, including oral, topical (nasal, transdermal, intraocular), intravesical, intrathecal, enteral, pulmonary, intralymphatic, intracavity, vaginal, transurethral, ​​intradermal, otic, intramammary, buccal, orthotopic, intratracheal, intralesional, transdermal, endoscopic, transmucosal, sublingual, and enteral administration.

[0125] Pharmaceutically acceptable additives and / or excipients for use in combination with the compositions of the present invention are known to those of skill in the art and are selected based on a number of factors: the particular compound and drug used and its(their) concentration, stability and intended bioavailability; the subject, its age, size and general condition; and the route of administration.Pharmaceutically acceptable excipients for use in the pharmaceutical compositions described herein are known to those skilled in the art and are described in The Chemotherapy Source Book (Williams & Wilkens Publishing), The Handbook of Pharmaceutical Excipients, (American Pharmaceutical Association, Washington, DC and The Pharmaceutical Society of Great Britain, London, England, 1968), Modern Pharmaceutics, (G. Banker et al., eds., 3rd ed.) (Marcel Dekker, Inc., New York, NY, 1995), The Pharmacological Basis of Therapeutics, (Goodman & Gilman, McGraw Hill Publishing), Pharmaceutical Dosage Forms, (H. Lieberman et al., eds.) (Marcel Dekker, Inc., New York, NY, 1980), Remington's Pharmaceutical Sciences (A. Gennaro, ed., 19th ed.) (Mack Publishing, Easton, PA, 1995), The United States Pharmacopeia 24, The National Formulary 19, (National Publishing, Philadelphia, PA, 2000) and AJ Spiegel et al., Use of Nonaqueous Solvents in Parenteral Products, Journal of Pharmaceutical Sciences, Vol. 52, No. 10, pp. 917-927 (1963).

[0126] Formulations for certain pentaaza macrocyclic ring complexes are described, for example, in U.S. Patent Nos. 5,610,293, 5,637,578, 5,874,421, 5,976,498, 6,084,093, 6,180,620, 6,204,259, 6,214,817, 6,245,758, 6,395,725, and 6,525,041, each of which is incorporated herein by reference in its entirety.

[0127] The pharmaceutical compositions comprising the pentaazamacrocycles may further comprise one or more additional pharmaceutically active ingredients. Suitable pharmaceutically active agents that may be included in the compositions according to the present invention include, for example, antiemetics, anesthetics, antihypertensives, antianxiety agents, anticoagulants, anticonvulsants, hypoglycemic agents, decongestants, antihistamines, antitussives, antineoplastic agents, beta-blockers, anti-inflammatory agents, antipsychotics, nootropics, cholesterol-lowering agents, antiobesity agents, autoimmune disorders, antiapathy agents, antibacterial and antifungal agents, hypnotics, antiparkinsonian agents, antialzheimer's agents, antibiotics, antidepressants, and antiviral agents. The individual components of such combinations may be administered separately or in combination with pharmaceutical formulations, sequentially or simultaneously.

[0128] In yet another embodiment, a kit containing a pentaazamacrocyclic ring complex for treating a condition can be provided. For example, the kit can include a first vessel or container containing therein a formulation comprising the pentaazamacrocyclic ring complex in aqueous solution, e.g., an oral or injectable formulation of the pentaazamacrocyclic ring complex. The kit can further include a label or other instructions for administration of the active agent, recommended dosage, duration and administration regimen, precautions, a list of potential drug-drug interactions, and other relevant instructions, such as a label indicating a treatment regimen (e.g., dosage, frequency of administration, etc.) corresponding to any of the treatment regimens described herein.

[0129] Combination treatment with cancer therapy In certain embodiments, the aqueous solution containing the pentaaza macrocyclic complex can be administered in combination with another cancer therapy to provide therapeutic treatment, for example, the pentaaza macrocyclic complex can be administered as part of a radiation or chemotherapy regimen.

[0130] In general, the temporal mode of administration of the pentaaza macrocyclic ring complex may depend, for example, on the type, nature, and / or duration of the specific radiation therapy selected. Other considerations may include factors such as the disease or disorder being treated and the severity of the disease or disorder; the activity of the specific compound used; the specific composition used; the age, weight, general health, sex, and diet of the subject; the time of administration, the route of administration, and the excretion rate of the specific compound used; the duration of treatment; and drugs used in combination with or simultaneously with the specific compound used. For example, in various embodiments, the compound may be administered before, during, and / or after administration of radiation therapy (e.g., before, during, or after exposure to radiation therapy including multiple exposures and / or doses and / or before, during, or after treatment with a series of radiation therapy including multiple exposures and / or doses). As another example, in various embodiments, the compound may be administered before, during, and / or after radiation exposure. If desired, the effective amount may be divided into multiple doses for administration purposes; such that a single-dose composition may contain such amounts or submultiples thereof to constitute the administered dose.

[0131] In some embodiments, for example, the pentaaza macrocyclic complex can be administered to a patient before or simultaneously with radiation exposure and / or chemotherapy dose. In other embodiments, for example, the compound is administered to a patient before, rather than after, radiation exposure and / or chemotherapy dose. In yet other embodiments, the pentaaza macrocyclic complex is administered to a patient at least 15 minutes, 30 minutes, 45 minutes, 60 minutes, 90 minutes, 180 minutes, 0.5 days, 1 day, 3 days, 5 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks or longer before radiation exposure and / or chemotherapy dose, for example, an initial radiation exposure in a radiation treatment series, or before another dose or dose fraction in a series of multiple doses or dose fractions of radiation. In yet other embodiments, for example, the pentaaza macrocycle complex is administered to the patient after radiation exposure and / or a chemotherapy dose; thus, for example, the compound is administered for up to 15 minutes, 30 minutes, 45 minutes, 60 minutes, 90 minutes, 180 minutes, 0.5 days, 1 day, 3 days, 5 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks or longer after radiation exposure, which may be a dose or dose fraction in multiple doses of radiation therapy, or may be a single or final dose or dose fraction in radiation therapy, or after a chemotherapy dose.

[0132] In some embodiments, a course of radiation therapy comprises multiple radiation doses or dose fractions administered over a predetermined period of time, such as hours, weeks, days, and even months, where the multiple radiation doses or dose fractions are similar or vary. That is, a course of radiation therapy can comprise the administration of a series of multiple radiation doses or dose fractions. In some embodiments, the pentaaza macrocyclic complex can be administered before one or more radiation doses or dose fractions in the course of radiation, for example, before each radiation dose or dose fraction, or before some radiation doses or dose fractions. Furthermore, the administration of the pentaaza macrocyclic complex during the course of radiation therapy can be selected to enhance the cancer treatment effect of radiation therapy. In some embodiments, the pentaaza macrocyclic complex can be administered within a predetermined period of time, such as within the above-mentioned predetermined period, before or after each dose or dose fraction. In another embodiment, the pentaaza macrocyclic complex is administered within a predetermined period of time only before or after a dose or dose fraction.

[0133] The appropriate total dose to be provided during a course of treatment can be determined by the treatment provided, the patient's physical characteristics and other factors, and the dose fractions provided can be determined as well. In some embodiments, the dose fraction of radiation administered to a patient can be at least 1.8 Gy, for example, at least 2 Gy, and even at least 3 Gy, for example, at least 5 Gy, and even at least 6 Gy. In yet another embodiment, the dose fraction of radiation administered to a patient can be at least 10 Gy, for example, at least 12 Gy, and even at least 15 Gy, for example, at least 18 Gy, and even at least 20 Gy, for example, at least 24 Gy. Generally, the dose fraction of radiation administered to a patient does not exceed 54 Gy. The total dose (i.e., the sum of all dose fractions) administered during a course of treatment can be at least 20 Gy, at least 30 Gy, at least 40 Gy, at least 50 Gy, at least 60 Gy, and / or at least 70 Gy. For example, the total dose can be in the range of 50 Gy to 75 Gy, e.g., in the range of 60 Gy to 72 Gy. Furthermore, in certain embodiments, the dose fraction delivered to a subject can refer to the amount delivered to a particular target region of the subject, e.g., a tumor region, although it should be noted that other tumor regions or surrounding tissues may be exposed to more or less radiation than specified by the nominal dose fraction.

[0134] For example, in some embodiments, the total dose of radiation provided during a course of treatment can be provided by a hypofractionated radiation therapy method, which typically involves providing a relatively high dose fraction administered over a relatively small number of sessions compared to a low dose fractionation method.Examples of such radiation therapy can include, but are not limited to, stereotactic radiosurgery (SRS), which typically refers to a single-fraction treatment that targets intracranial and spinal cord targets, and stereotactic body radiotherapy (SBRT), which typically refers to a multi-fraction treatment of targets such as intracranial and spinal cord targets and extracranial targets such as lung, liver, head and neck, pancreas and prostate.For example, in some embodiments of a hypofractionated radiation therapy method, the total dose of radiation provided during a course of treatment can be divided into less than 10 fractions, for example, less than 8 fractions, less than 6 fractions, less than 5 fractions, less than 4 fractions, less than 3 fractions, less than 2 fractions, and even can be provided in only one administration (single fraction). For example, in certain embodiments, the total dose of radiation provided during a course of treatment may be fractionated into 1 to 10 fractions, e.g., 1 to 6 fractions, and even 1 to 5 fractions, e.g., 2 to 5 fractions or even 2 to 4 fractions. As yet another example, a hypofractionated radiation therapy regimen may include fractionating the total dose of radiation provided during a course of treatment into dose fractions that are at least 10% (1 / 10) of the total dose, e.g., at least 12.5% ​​(1 / 8) of the total dose, at least 16% (about 1 / 6) of the total dose, at least 20% (1 / 5) of the total dose, at least 25% (1 / 4) of the total dose, at least 30% (1 / 3) of the total dose, at least 50% of the total dose, and / or at least 100% of the total dose may be administered in a single administration (single fraction). For example, in certain embodiments, the total dose of radiation provided during a course of treatment can be divided into fractions that provide 10% to 100% of the total dose in each fraction, e.g., 16% to 100% of the total dose, and even 20% to 100% of the total dose, e.g., 20% to 50% of the total dose or even 25% to 50% of the total dose.For example, the dose fraction size can be at least 5 Gy, e.g., at least 6 Gy, at least 8 Gy, at least 10 Gy, at least 12 Gy, and even at least 15 Gy, e.g., at least 18 Gy, and even at least 20 Gy, e.g., at least 24 Gy, and typically not more than 54 Gy, e.g., less than 40 Gy and even less than 30 Gy. In certain embodiments, the dose fraction size can range from 5 Gy to 30 Gy, e.g., 6 Gy to 28 Gy, and even 8 Gy to 25 Gy. Furthermore, in certain embodiments, the dose fractions can be administered no more than three times per day, and even no more than twice per day, e.g., no more than once per day, on consecutive or non-consecutive days and / or some combination thereof, over a period of several days and up to several weeks, e.g., 1 to 15 days, 1 to 12 days, 1 to 10 days, 1 to 5 days, and even 1 to 3 days. Typically, the dose fractions making up the entire course of treatment are administered within 20 days, 15 days, 10 days, 5 days, and even 3 days.

[0135] As yet another example, in some embodiments, the total dose of radiation provided during a course of treatment can be provided by a radiotherapy regimen that provides a relatively low dose fraction administered over a relatively large number of sessions, for example, compared to a hypofractionated radiation regimen. Examples of such low-fractionated radiation therapy can include, but are not limited to, intensity-modulated radiation therapy (IMRT) and image-guided radiation therapy (IGRT), which typically include three-dimensional conformal radiation therapy (3D-CRT) to adapt the administered radiation to the target volume. For example, in some embodiments of such a radiotherapy regimen, the total dose of radiation provided during a course of treatment can be divided into at least 15 fractions, for example, at least 18 fractions, at least 20 fractions, at least 22 fractions, at least 25 fractions, at least 28 fractions, at least 30 fractions, at least 32 fractions, at least 35 fractions, and even at least 38 fractions, while the total number of fractions is typically less than 50, for example, less than 45, and even less than 42. For example, in certain embodiments, the total dose of radiation provided during a course of treatment can be fractionated into 15 to 38 fractions, e.g., 20 to 38 fractions, and even 20 to 35 fractions, e.g., 25 to 35 fractions. As yet another example, a radiation therapy regimen can include fractionating the total dose of radiation provided during a course of treatment into dose fractions that are 7% (1 / 15) or less of the total dose provided during a course of treatment, e.g., 6% (1 / 18) or less of the total dose, 5% (1 / 20) or less of the total dose, 4.5% (1 / 22) or less of the total dose, 4% (1 / 25) or less of the total dose, 3.6% (1 / 28) or less of the total dose, 3.3% (1 / 30) or less of the total dose, 3.1% (1 / 32) or less of the total dose, 2.8% (1 / 35) or less of the total dose, and even 2.6% (1 / 38) or less of the total dose. For example, in certain embodiments, the total dose of radiation provided during a course of treatment may include fractionation into fractions that provide 2.5% to 8% of the total dose in each fraction, e.g., 2.8% to 5%, and even 2.8% to 4% of the total dose.For example, the dose fraction size can be less than 5 Gy, e.g., less than 4 Gy, less than 3.5 Gy, less than 3 Gy, less than 2.8 Gy, and even less than 2.5 Gy, e.g., less than 2.3 Gy, and even less than 2 Gy, e.g., less than 1.8 Gy, and typically is at least 0.5 Gy, e.g., at least 1 Gy and even at least 1.5 Gy. In some embodiments, the dose fraction size can be administered in the range of 1.5 Gy to 4.5 Gy, e.g., 1.8 Gy to 3 Gy, and even 2 Gy to 2.5 Gy. Further, in some embodiments, dose fractions can be administered no more than three times per day, and even no more than two times per day, e.g., no more than once per day, on consecutive days or non-consecutive days and / or some combination thereof (e.g., consecutive weekdays), and in some embodiments over a period of days to weeks and even months, e.g., up to 3 weeks, up to 5 weeks, up to 6 weeks, up to 8 weeks, and even up to 10 weeks, e.g., in the range of 3 weeks to 10 weeks, or even in the range of 5 weeks to 8 weeks. For example, the dose fractions making up the entire course of treatment can be administered within 12 weeks, e.g., within 10 weeks, and even within 8 weeks.

[0136] In yet another embodiment, the total dose of radiation provided by the irradiation method is selected to provide appropriate treatment for cancer, regardless of whether it is a relatively high dose fractionation method or a relatively low dose fractionation method or other method as described above.The total dose can also be provided by the specific dose fractionation fraction administered, along with other factors.For example, in certain embodiments, a relatively high total dose can be administered as a relatively low individual dose fraction.In some embodiments, the total dose provided over the course of treatment (i.e., the sum of the dose fractions administered) is at least 50Gy, for example, at least 55Gy, at least 58Gy, at least 60Gy, at least 65Gy, at least 68Gy, at least 70Gy, at least 72Gy, and even at least 75Gy.In certain embodiments, the total dose does not exceed 80Gy, for example, does not exceed 78Gy, and even does not exceed 75Gy.For example, the total dose is in the range of 50Gy to 75Gy, for example, 55Gy to 75Gy, and even 60Gy to 70Gy.

[0137] In yet another embodiment, the pentaaza macrocyclic complexes can be administered as part of a course of treatment that includes, for example, administration of a platinum-based chemotherapy agent (e.g., cisplatin). In chemotherapy, chemotherapeutic agents are administered to a patient to kill cancer cells or control the growth of cancer cells. A typical course of chemotherapy includes one or more doses of one or more chemotherapeutic agents, which can be administered over the course of days, weeks, and even months. Chemotherapeutic agents include antineoplastic alkylating agents such as nitrogen mustards (e.g., cyclophosphamide, chlorambucil), nitrosoureas (e.g., n-nitroso-n-methylurea, carmustine, semustine), tetrazines (e.g., dacarbazine, mitozolimide), aziridines (e.g., thiotepa, mitomycin); antimetabolites such as antifolates (e.g., methotrexate and pemetrexed), fluoropyrimidines (e.g., fluorouracil, capecitabine), anthracyclines (e.g., doxorubicin, daunorubicin), The therapeutic agent may include at least one of: deoxynucleoside analogs (e.g., cytarabine, gemcitabine, decitabine) and thiopurines (e.g., thioguanine, mercaptopurine); anti-microtubule agents, such as taxanes (e.g., paclitaxel, docetaxel); topoisomerase inhibitors (e.g., etoposide, doxorubicin, mitoxantrone, teniposide); antitumor antibiotics (e.g., bleomycin, mitomycin); and platins (e.g., cisplatin, carboplatin, oxaliplatin). For example, the chemotherapeutic agent may be selected from the group consisting of retinoic acid, arsenic trioxide, azacitidine, azathioprine, bleomycin, carboplatin, capecitabine, cisplatin, chlorambucil, cyclophosphamide, cytarabine, daunorubicin, docetaxel, doxifluridine, doxorubicin, epirubicin, epothilone, etoposide, fluorouracil, gemcitabine, hydroxyurea, idarubicin, imatinib, mechlorethamine, mercaptopurine, methotrexate, mitoxantrone, oxaliplatin, paclitaxel, pemetrexed, teniposide, tiguanine, valrubicin, vinblastine, vincristine, vindesine, and vinorelbine.The administration of many chemotherapeutic agents is described in the "Physicians' Desk Reference" (PDR), for example, 1996 edition (Medical Economics Company, Montvale, NJ 07645-1742, USA).

[0138] In some embodiments, the chemotherapeutic agent comprises any one selected from the group consisting of platinum-based anticancer agents, such as cisplatin, carboplatin, oxaliplatin, nedaplatin, lobaplatin, heptaplatin, dicycloplatin, lipoplatin, LA-12 ((OC-6-43)-bis(acetato)(1-adamantylamine)aminedichloroplatinum(IV)), phosphaplatin, phenanthriplatin, ProLindac (AP5346), triplatin tetranitrate, picoplatin, satraplatin, piriplatin, and / or pharmaceutically acceptable salts thereof. An example of a suitable dose of a platinum-based anticancer agent is 10 mg / m 2 ~200mg / m 2 , e.g., 20 mg / m 2 ~100mg / m 2 The administration schedule of platinum-based anticancer drugs can be selected according to the intended treatment and the platinum-based anticancer drug provided.For example, in some embodiments, suitable administration schedules can include administering to patients once or twice a day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, or 1 month.

[0139] According to yet another embodiment, a method of treatment may involve the combination of a pentaaza macrocyclic complex with an immunotherapeutic agent, such as an immune checkpoint inhibitor, adoptive T cell transfer therapy, and / or a cancer vaccine, which may be administered for the treatment of cancer, and optionally as part of a course of treatment that also includes chemotherapeutic agents and / or radiation therapy. [Example]

[0140] The following non-limiting examples are provided to further illustrate aspects of the present invention. It should be understood by those skilled in the art that the techniques disclosed in the following examples represent approaches that the inventors have found to work well in practicing the invention, and therefore can be considered to constitute examples of modes for carrying out the invention. However, those skilled in the art should understand in light of the present invention that many changes can be made in the specific embodiments disclosed and still obtain the same or similar results without departing from the concept and scope of the invention.

[0141] Effect of addition order on precipitate formation in aqueous formulations Example 1 In this example, a manganese-containing coordination complex corresponding to the structural formula of GC4419 disclosed herein was combined with sodium chloride and sodium bicarbonate (as a buffering agent) to form aqueous formulations by adding these components in different orders, and the formulations were monitored to visually observe whether particles (precipitates) formed therein.

[0142] Specifically, a first set of formulations (Formulations A-1 and A-2) was prepared by adding 9 mg / mL of GC4419 to water (WFI) adjusted to a pH of 7.4-7.8, followed by (2) sodium chloride (0.9% NaCl), and finally (3) sodium bicarbonate (26 mM), optionally with water to achieve the desired concentration. A second set of formulations (Formulations B-1 and B-2) was prepared by adding 9 mg / mL of GC4419 to water (WFI) adjusted to a pH of 7.4-7.8, followed by (2) sodium bicarbonate (26 mM), and finally (3) sodium chloride (0.9% NaCl), optionally with water to achieve the desired concentration. A third set of formulations (Formulations C-1, C-2, and C-3) was then prepared in a manner similar to A-1 and A-2. That is, Formulations "A" and "C" differ from Formulation "B" in the order of addition of sodium bicarbonate relative to the sodium chloride added to the solution. Table 1 below shows the results for visible particles (precipitation) observed for each of the formulations. <788> Results for subvisible particles, assayed as described in and reported as particle counts >10 μM and >25 μM in diameter, are also shown. [Table 3]

[0143] Thus, as can be seen from the above, formulations "A" and "C," in which sodium chloride was added before sodium bicarbonate, showed no visible precipitation and very few invisible particles even after 9 and 12 months. However, simply reversing the order of addition, formulation "B," in which sodium bicarbonate was added before sodium chloride, resulted in observable precipitation and many invisible particles after 9 months, even though such precipitation was not observed immediately after preparation of the formulation (e.g., at 0 months).

[0144] The precipitate formed after 9 months in the "B" formulation was further analyzed to determine the chemical composition and physical characteristics of the precipitate, including polarized light microscopy (PLM), elemental analysis using a field-emission scanning electron microscope (FESEM) equipped with an energy-dispersive X-ray spectrometer (EDS), electron backscatter diffraction (EBSD), X-ray fluorescence (XRF), and Raman microspectroscopy. Microscopic analysis determined that the precipitate contained crystals consistent in appearance with rhodochrosite (manganese carbonate, MnCO), and analysis of these crystals by Raman microspectroscopy confirmed this identity. Figure 5 is a photograph showing MnCO crystals obtained from the "B" formulation, as they appear in both simple polarized light (bottom left) and crossed polarized light (top right). Figure 6 shows the Raman spectrum of MnCO3 crystals from the "B" formulation (top spectrum-A) compared to a reference spectrum for rhodochrosite (second spectrum-B), a Raman spectrum collected from a sample of MnO2 (third spectrum-B), and a library reference spectrum of manganese sieve, Mn3O4 (bottom spectrum-D).

[0145] Furthermore, formulations manufactured in a manner similar to formulations "A" and "C," but with GC4419 concentrations of 3 mg / mL and 10 mg / mL (rather than 9 mg / mL in A-1, A-2, C-1, C-2, and C-3 above), also showed no precipitate upon visual inspection 9 or 12 months after their manufacture.

[0146] Example 2 In this example, MnCl2 was combined with sodium chloride and sodium bicarbonate (as a buffering agent) to form aqueous formulations with different orders of addition of these components to evaluate the effect of addition order on the precipitation of manganese from solution over time.

[0147] Specifically, a first set of formulations (Formulation 1—"Order, NaCl 1") was prepared by adding 0.026 mM, 0.26 mM, 2.6 mM, and 26 mM MnCl to water (WFI) adjusted to a pH of 7.4–7.8, followed by (2) sodium chloride (0.9% NaCl), and finally (3) sodium bicarbonate (26 mM). st A second set of formulations (Formulation 2 - "Order, NaHCO3 1") was prepared by adding 0.026 mM, 0.26 mM, 2.6 mM, and 26 mM MnCl2 to water (WFI) adjusted to a pH of 7.4-7.8, followed by (2) sodium bicarbonate (26 mM), and finally (3) sodium chloride (0.9% NaCl). st A third set of formulations (Formulation 3—"Vehicle") was prepared by adding 0.026 mM, 0.26 mM, 2.6 mM, and 26 mM MnCl to water (WFI) adjusted to a pH of 7.4–7.8, followed by (2) a solution containing sodium bicarbonate (26 mM) and sodium chloride (0.9% NaCl). Formulation 2 differs from Formulations 1 and 3 in that sodium bicarbonate is added before sodium chloride in Formulation 2, as opposed to being added simultaneously (Formulation 3) or after (Formulation 1) with sodium chloride.

[0148] The amount of manganese precipitate formed 1 and 6 days after formulation of formulations (1) to (3) was assessed by an ICP-MS storage stability assay, which involved filtering the formulations through a 0.45 micrometer filter, washing the filter with water at pH 8.0, dissolving the filter with nitric acid, and performing inductively coupled mass spectrometry (ICP-MS) to detect the manganese content of any precipitates.

[0149] 1-2, the results for Formulations (1)-(3) can be seen one day after preparation. Specifically, it can be seen that very low concentrations of MnCl2 (0.026 ppm or 0.26 mM MnCl2) resulted in very little or negligible amounts of manganese precipitation (2.20 ppm or 1 ppm detected Mn) at one day, while Formulations (1)-(3) solutions with 2.6 mM or 26 mM MnCl2 provided significantly different amounts of precipitation depending on the order of addition of the aqueous solution components. In particular, formulations (1) and (3) showed 179.84 ppm Mn and 104.48 ppm Mn for the 2.6 mM MnCl2 solution, and 1179.05 ppm Mn and 974.93 ppm Mn for the 26 mM MnCl2 solution at 1 day, while formulation (2), which added sodium bicarbonate first, showed an approximately 177% increase in measured Mn at 1 day, showing 319.14 ppm Mn for the 2.6 mM MnCl2 solution and 2250 ppm Mn for the 26 mM MnCl2 solution. These results are shown in chart form in Figure 1 and graphically in Figure 2.

[0150] Similarly, Figures 3-4 show the results for Formulations (1)-(3) 6 days after preparation. As at Day 1, it can be seen that extremely low concentrations of MnCl2 (0.026 ppm or 0.26 mM MnCl2) resulted in very little or negligible amounts of manganese precipitation (1 ppm of detected Mn) at Day 1. However, Formulations (1)-(3) solutions with 2.6 mM or 26 mM MnCl2 provided significantly different amounts of precipitation depending on the order of addition of the aqueous solution components. In particular, formulations (1) and (3) showed 36.08 ppm Mn and 44.02 ppm Mn for the 2.6 mM MnCl2 solution, and 1162 ppm Mn and 926.31 ppm Mn for the 26 mM MnCl2 solution at 6 days, while formulation (2), to which sodium bicarbonate was added first, showed an approximately 750% increase in measured Mn at 6 days, showing 270.63 ppm Mn for the 2.6 mM MnCl2 solution and 2250 ppm Mn for the 26 mM MnCl2 solution. These results are shown in chart form in Figure 3 and graphically in Figure 4.

[0151] Thus, the results indicate that a relatively small amount of Mn-containing component in an aqueous formulation can result in the formation of a significant amount of precipitate when sodium bicarbonate is added to the Mn-containing component before the addition of sodium chloride. That is, sodium chloride provides a protective effect to reduce precipitate formation when added to a solution with the Mn-containing component before or simultaneously with sodium bicarbonate to provide an excess of chloride ions compared to the dianion produced by sodium bicarbonate.

Claims

1. 1. A method for preparing an aqueous formulation of a manganese-containing coordination complex, the formulation comprising a manganese-containing coordination complex, a chloride anion, and a dianion, the method comprising: combining a source of a manganese-containing coordination complex with a source of chloride anions in an aqueous solution; and providing a dianion source in aqueous solution simultaneously or subsequently to combining a chloride anion source and a manganese-containing coordination complex source in aqueous solution to form an aqueous formulation. wherein the amount of chloride anion source combined with the manganese-containing coordination complex is sufficient to provide a chloride ion concentration in the aqueous formulation that exceeds the dianion concentration in the aqueous formulation; The manganese-containing coordination complex has the formula: 【Chemistry 1】 is a compound represented by The method.

2. 10. The method of claim 1, wherein the Mn(II)-containing component is present in the source of manganese-containing coordination complex in a weight ratio of Mn(II)-containing component to manganese-containing coordination complex that is in the range of 1:100,000 to 1:100, and / or in the range of 1:75,000 to 1:1,000, and / or in the range of 1:50,000 to 1:5,000, and / or in the range of 1:15,000 to 1:8,000.

3. 3. The method of claim 1, wherein the chloride anion source comprises a salt capable of forming chloride anions in aqueous solution.

4. 4. The method of claim 1, wherein the chloride anion source comprises at least one selected from the group consisting of sodium chloride, potassium chloride, calcium chloride, and magnesium chloride.

5. 5. The method of any one of claims 1 to 4, comprising adding a chloride anion source in an amount sufficient to provide a chloride anion concentration in the aqueous formulation of at least 100 mM, at least 110 mM, at least 115 mM, at least 120 mM, at least 130 mM, at least 145 mM and / or at least 150 mM, and not more than 1000 mM, not more than 200 mM, not more than 180 mM, not more than 175 mM, not more than 160 mM and / or not more than 155 mM.

6. The method of any one of claims 1 to 5, wherein the dianion source comprises a bicarbonate salt.

7. 7. The method of any one of claims 1 to 6, comprising adding a dianion source in an amount sufficient to provide a dianion concentration in the aqueous formulation of at least 0.1 mM, at least 0.25 mM, at least 1 mM, and / or at least 2.5 mM, and no more than 26 mM, no more than 15 mM, and / or no more than 10 mM.

8. 8. The method according to any one of claims 1 to 7, wherein the dianion source is added in an amount sufficient for a pH range within a pH range of 7 to 10 and / or within a pH range of 7.5 to 9.

9. 9. The method of any one of claims 1 to 8, wherein the concentration of chloride anion in the aqueous formulation exceeds the concentration of dianion in the formulation by a concentration ratio of chloride anion to dianion in mol / L of at least 10:1, at least 100:1, at least 250:1, at least 500:1, at least 750:1, at least 1000:1, at least 5000:1, and / or at least 10,000:

1.

10. 10. The method of any one of claims 1 to 9, wherein at least 75 mol%, at least 85 mol%, at least 90 mol%, at least 95 mol%, at least 98 mol%, at least 99 mol% and / or the total molar amount of the dianion source added to form the aqueous formulation is added simultaneously or subsequently to combining the chloride anion source and the manganese-containing coordination complex.

11. 11. The method of any one of claims 1 to 10, wherein the manganese-containing coordination complex is combined with the chloride anion source in aqueous solution, and then the dianion source is added to the aqueous solution containing the chloride anions and manganese-containing coordination complex for at least 30 seconds, at least 1 minute, at least 5 minutes, at least 10 minutes, at least 30 minutes, and / or at least 1 hour.

12. 12. The method of any one of claims 1 to 11, further comprising adjusting the aqueous solution to a pH of at least 8 before combining therewith a manganese-containing coordination complex and a chloride anion source.

13. 13. The method of any one of claims 1 to 12, wherein the aqueous formulation comprises a buffered formulation for parenteral administration of a manganese-containing coordination complex, said buffered formulation having a physiological level of sodium chloride.

14. 14. The method of any one of claims 1 to 13, wherein the concentration of the manganese-containing coordination complex in the aqueous formulation is at least 2 mM, at least 6 mM, at least 18 mM, at least 20 mM and / or at least 40 mM.

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