Production of trans-[tetrachlorobis(1H-indazole)ruthenate(III)] and its compositions

Abstract

To provide the manufacture of trans-[tetrachlorobis(1H-indazole)ruthenate (III)] and compositions thereof.SOLUTION: The present invention relates to the preparation of compositions comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)]. Synthesis and formulation preparation are detailed. Impurity profiles are also discussed. Compositions herein are useful for anti-cancer applications. According to an embodiment, the invention provides a composition that comprises sodium trans-[tetrachlorobis(1H-indazole)ruthenate (III)], and cesium. In some embodiments, the compositions can further comprise RuIIICl3(Hind)2(H2O), RuIIICl3(Hind)2(CH3CN), and RuIIICl3(Hind) (HN=C(Me) ind).SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] Cross - Reference to Related Applications This application claims the benefit of U.S. Provisional Patent Application No. 62 / 501,984, filed on May 5, 2017. The entire disclosure of the provisional application is incorporated herein by reference.

[0002] Field of the Invention The present invention generally relates to chemical synthesis, and more particularly to a method for preparing an alkali metal salt of trans - [tetrachlorobis(1H - indazole)ruthenate(III)].

Background Art

[0003] Several methods for the preparation of trans - [tetrachlorobis(1H - indazole)ruthenate(III)] sodium (also known as KP1339, NKP - 1339, IT - 139, and Na[Ru III Cl4(Hind)2]) exist in the literature. For example, W. Peti et al, Eur. J. Inorg. Chem. 1999, 1551 - 1555 disclose the following synthetic scheme.

Chemical Formula

[0004] In this method, due to the limited solubility of tetramethylammonium chloride salt, a large amount of solvent is consequently required. Furthermore, there are concerns about toxicity regarding the use of tetramethylammonium salts. U.S. Patent No. 8,362,266 describes an additional process. This process is a method for preparing the compound M - trans - [tetrachlorobis(1H - indazole)ruthenate(III)] (where M is an alkali metal cation), comprising: (1) reacting trans - [tetrachlorobis(1H - indazole)ruthenate(III)] indazolium in an aqueous solution or in a mixture of water and a water - soluble first organic solvent with an inorganic salt of the alkali metal cation M to form the compound M - trans - [tetrachlorobis(1H - indazole)ruthenate(III)] and an inorganic salt of indazole; and (2) extracting the indazole from the M - trans - [tetrachlorobis(1H - indazole)ruthenate(III)] using a second organic solvent that is substantially water - insoluble. This method is summarized in the following scheme. [Chemical formula]

[0005] Although the above - mentioned method is effective, the extraction step and the associated holding time are required, which may limit the effective batch size. Also, the purity of the compound is directly related to the length of time the compound is in a basic aqueous environment. The overall yield in this method is 20 - 35%. Therefore, a method that does not utilize an extraction process, avoids an aqueous basic environment, has a high yield, and produces the compound at a high purity level is highly desirable. Furthermore, a methodology that avoids extraction and large amounts of organic solvents is also desirable. A methodology that focuses primarily on precipitation and then filtration is considered to meet this requirement. [Prior Art Documents] [Patent Documents]

[0006] [Patent Document 1] U.S. Patent No. 8,362,266 Specification

Non-Patent Literature

[0007]

Non-Patent Literature 1

Brief Description of the Drawings

[0008]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Modes for Carrying Out the Invention

[0009] Detailed Description of Certain Embodiments of the Present Invention 1. General Description As described herein, the present invention relates to trans-[tetrachlorobis(1H-ind Provided is a method for preparing an alkali metal salt of [[dazole)ruthenium(III) acid]]. Such compounds include compounds of formula I. [Chemical formula] [wherein M is an alkali metal cation]

[0010] The present invention provides synthetic intermediates useful for the preparation of such compounds.

[0011] The present invention also provides a method for the preparation of cesium [trans-tetrachlorobis(1H-indazole)ruthenate(III)] shown by the following formula I-a. [Chemical formula]

[0012] The present invention also provides a method for the preparation of sodium [trans-tetrachlorobis(1H-indazole)ruthenate(III)] shown by the following formula I-b. [Chemical formula]

[0013] 2. Definitions trans-[tetrachlorobis(1H-indazole)ruthenate(III)] sodium Lithium, KP1339, NKP-1339, IT-139, and Na[Ru III Cl4(Hind)2] all correspond to the same compound (formula I-b), and it should be understood that these terms can be used interchangeably.

[0014] As used herein, the term amorphous refers to an amorphous solid lacking long-range order.

[0015] The compounds of the present invention include those generally described above, and the compounds of the present invention are further illustrated by the embodiments, sub-embodiments, and species disclosed herein. As used herein, unless otherwise indicated, the following definitions apply. For the purposes of the present invention, chemical elements are identified according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75 th Ed. In addition, general principles of organic chemistry are described in “Organic Chemistry”, Thomas Sorrell, University Science Books, Sausalito: 1999, and “March’s Advanced Organic Chemistry”, 5 th Ed., Ed.: Smith, M.B. and March, J., John Wiley & Sons, New York: 2001.

[0016] As used herein, the term “aliphatic” or “aliphatic group” refers to a hydrocarbon moiety that can be linear (i.e., unbranched), branched, or cyclic (including fused, bridged, and spiro-fused polycyclic), and may be completely saturated or contain one or more unsaturated (but not aromatic) units. Unless otherwise specified, aliphatic groups contain from 1 to 20 carbon atoms. In some embodiments, aliphatic groups contain from 1 to 10 carbon atoms. In other embodiments, aliphatic groups contain from 1 to 8 carbon atoms. In still other embodiments, aliphatic groups contain from 1 to 6 carbon atoms, and in yet other embodiments, aliphatic groups contain from 1 to 4 carbon atoms. Aliphatic groups include, but are not limited to, straight-chain or branched alkyl, alkenyl, and alkynyl groups, and hybrids thereof, such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl, or (cycloalkyl)alkenyl.

[0017] The term "heteroatom" means one or more of oxygen, sulfur, nitrogen, phosphorus, or silicon. This heteroatom includes any oxidation form of nitrogen, sulfur, phosphorus, or silicon; any quaternized form of any basic nitrogen, or a replaceable nitrogen in a heterocyclic ring (such as =N- in 3,4-dihydro-2H-pyrrolyl, -NH- in pyrrolidinyl, or =N(R † ))-.

[0018] As used herein, the term "unsaturated" means that a moiety has one or more unsaturation units.

[0019] As used herein, the term "divalent, saturated or unsaturated, straight-chain or branched C 1-12 hydrocarbon chain" refers to divalent alkylene, alkenylene, and alkynylene chains that are straight-chain or branched as defined herein.

[0020] The term "aryl", used alone or as part of a larger moiety such as in "aralkyl", "aralkoxy", or "aryloxyalkyl", refers to monocyclic, bicyclic, and tricyclic ring systems having a total of 5 to 14 ring members, wherein at least one ring in the system is aromatic and each ring in the system contains 3 to 7 ring members. The term "aryl" may be used interchangeably with the term "aryl ring".

[0021] As described herein, the compounds of the present invention may contain a "optionally substituted" moiety. Generally, the term "substituted", whether or not preceded by the term "optionally", means that one or more hydrogens in the indicated moiety are replaced by suitable substituents. Unless otherwise indicated, an "optionally substituted" group can have suitable substituents at each of its substitutable positions, and when two or more positions in any given structure can be substituted by two or more substituents selected from the designated group, the substituents may be the same or different at each position. The combinations of substituents contemplated by the present invention are preferably combinations that result in the formation of stable or chemically feasible compounds. As used herein, the term "stable" refers to a compound that, when subjected to the conditions that allow for its production, detection, and in certain embodiments its recovery, purification, and use for one or more of the purposes disclosed herein, is not substantially modified.

[0022] Monovalent substituents on a substitutable carbon atom of an "optionally substituted" group are independently halogen; -(CH2) 0-4 R ○ ; -(CH2) 0-4 OR ○ ; -O-(CH2) 0-4 C(O)OR ○ ; -(CH2) 0-4 CH(OR ○ )2; -(CH2) 0-4 SR ○ ; -(CH2) 0-4 Ph (optionally substituted with R ○ ); -(CH2) 0-4 O(CH2) 0-1 Ph (optionally substituted with R ○ ); -CH=CHPh (optionally substituted with R ○ ); -NO2; -CN; -N3; -(CH2) 0-4 N(R ○ )2; -(CH2) 0-4 N(R ○ )C(O)R ○ ; -N(R ○)C(S)R ○ ;-(CH2) 0-4 N(R ○ )C(O)NR ○ 2;-N(R ○ )C(S)NR ○ 2;-(CH2) 0-4 N(R ○ )C(O)OR ○ ;-N(R ○ )N(R ○ )C(O)R ○ ;-N(R ○ )N(R ○ )C(O)NR ○ 2;-N(R ○ )N(R ○ )C(O)OR ○ ;-(CH2) 0-4 C(O)R ○ ;-C(S)R ○ ;-(CH2) 0-4 C(O)OR ○ ;-(CH2) 0-4 C(O)SR ○ ;-(CH2) 0-4 C(O)OSiR ○ 3;-(CH2) 0-4 OC(O)R ○ ;-OC(O)(CH2) 0-4 SR-、SC(S)SR ○ ;-(CH2) 0-4 SC(O)R ○ ;-(CH2) 0-4 C(O)NR ○ 2;-C(S)NR ○ 2;-C(S)SR ○ ;-SC(S)SR°、-(CH2) 0-4 OC(O)NR ○ 2;-C(O)N(OR ○ )R ○ ;-C(O)C(O)R ○ ;-C(O)CH2C(O)R ○ ;-C(NOR ○ )R ○ ;-(CH2) 0-4 SSR ○ ;-(CH2) 0-4 S(O)2R ○ ;-(CH2) 0-4 S(O)2OR○ ;-(CH2) 0-4 OS(O)2R ○ ;-S(O)2NR ○ 2;-(CH2) 0-4 S(O)R ○ ;-N(R ○ )S(O)2NR ○ 2;-N(R ○ )S(O)2R ○ ;-N(OR ○ )R ○ ;-C(NH)NR ○ 2;-P(O)2R ○ ;-P(O)R ○ 2;-OP(O)R ○ 2;-OP(O)(OR ○ )2;SiR ○ 3;-(C 1-4 linear or branched alkylene)O-N(R ○ )2; or -(C 1-4 linear or branched alkylene)C(O)O-N(R ○ )2, and at this time, each R ○ may be substituted as defined below and independently is hydrogen, C 1-6 aliphatic, -CH2Ph, -O(CH2) 0-1 Ph, or a 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, regardless of the above definition, two independent occurrences of R ○ together with intervening atom(s) form a 3- to 12-membered saturated, partially unsaturated, or aryl monocyclic or bicyclic ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, which may be substituted as defined below.

[0023] R ○ (or the ring formed by combining the presence of two independent R ○ with intervening atom) the monovalent substituents on are independently halogen, -(CH2) 0-2 R ● 、-(haloR ● )、-(CH2) 0-2OH, -(CH2) 0-2 OR ● , -(CH2) 0-2 CH(OR ● )2; -O(haloR ● ), -CN, -N3, -(CH2) 0-2 C(O)R ● , -(CH2) 0-2 C(O)OH, -(CH2) 0-2 C(O)OR ● , -(CH2) 0-2 SR ● , -(CH2) 0-2 SH, -(CH2) 0-2 NH2, -(CH2)0 -2 NHR ● , -(CH2) 0-2 NR ● 2, -NO2, -SiR ● 3, -OSiR ● 3, -C(O)SR ● 、 -(C 1-4 linear or branched alkylene)C(O)OR ● , or -SSR ● wherein each R ● is unsubstituted or, when preceded by "halo", substituted with only one or more halogens, and independently is selected from C 1-4 aliphatic, -CH2Ph, -O(CH2) 0-1 Ph, or a 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Divalent substituents on the saturated carbon atoms of such R ○ include =O and =S.

[0024] Divalent substituents on the saturated carbon atoms of an optionally substituted group include =O, =S, =NNR * 2, =NNHC(O)R * , =NNHC(O)OR * , =NNHS(O)2R * , =NR * , =NOR * , -O(C(R * 2))2-3 O- or -S(C(R * 2)) 2-3 S- is included, and at this time, each independent existence of R * is selected from hydrogen, C 1-6 aliphatic (which may be substituted as defined below), or an unsubstituted 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. As the divalent substituent bonded to the substitutable carbon adjacent to the "optionally substituted" group, -O(CR * 2) 2-3 O- is included, and at this time, each independent existence of R * is selected from hydrogen, C 1-6 aliphatic (which may be substituted as defined below), or an unsubstituted 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. The tetravalent substituent bonded to the substitutable methylene carbon adjacent to the "optionally substituted" group is a dicobalt hexacarbonyl cluster, and when illustrated together with the methylene having it,

Chemical formula

[0025] R * Suitable substituents on the aliphatic group of are halogen, -R ● , -(haloR ● ), -OH, -OR ● , -O(haloR ● ), -CN, -C(O)OH, -C(O)OR ● , -NH2, -NHR ● , -NR ● 2, or -NO2 is included, and at this time, each R ● is unsubstituted or, when "halo" precedes, is substituted only by one or more halogens, and independently, C 1-4 aliphatic, -CH2Ph, -O(CH2) 0-1It is a 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms independently selected from Ph, or nitrogen, oxygen, or sulfur.

[0026] Suitable substituents on the nitrogen of the "optionally substituted" group include -R † , -NR † 2, -C(O)R † , -C(O)OR † , -C(O)C(O)R † , -C(O)CH2C(O)R † , -S(O)2R † , -S(O)2NR † 2, -C(S)NR † 2, -C(NH)NR † 2, or -N(R † )S(O)2R † , and at this time, each R † is independently hydrogen, C 1-6 aliphatic (optionally substituted as defined below), unsubstituted -OPh, or a 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, regardless of the above definition, two independent occurrences of R † together with intervening atom(s), form a 3- to 12-membered saturated, partially unsaturated, or aryl monocyclic or bicyclic ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0027] Suitable substituents on the aliphatic group of R † are independently halogen, -R ● , -(haloR ● ), -OH, -OR ● , -O(haloR ● ), -CN, -C(O)OH, -C(O)OR ● , -NH2, -NHR ● , -NR ● 2, or -NO2, and at this time, each R ●is unsubstituted or substituted with only one or more halogens when "halo" precedes, and independently, C 1-4 aliphatic, -CH2Ph, -O(CH2) 0-1 Ph, or a 5- to 6-membered saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0028] Protected hydroxyl groups are well known in the art and are described in detail in Protecting Groups in Organic Synthesis, T.W. Greene and P.G.M. Wuts, 3 rdIt is described in the 1999 edition of John Wiley & Sons (the whole of which is incorporated herein by reference). Examples of suitable protecting hydroxyl groups further include, but are not limited to, esters, carbonates, sulfonate allyl ethers, ethers, silyl ethers, alkyl ethers, arylalkyl ethers, and alkoxyalkyl ethers. Examples of suitable esters include formate, acetate, propionate, pentanoate, crotonate, and benzoate. Specific examples of suitable esters include formate, benzoylformate, chloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate, 4,4-(ethylenedithio)pentanoate, pivaloate (trimethylacetate), crotonate, 4-methoxy-crotonate, benzoate, p-benylbenzoate, 2,4,6-trimethylbenzoate. Examples of carbonates include 9-fluorenylmethyl, ethyl, 2,2,2-trichloroethyl, 2-(trimethylsilyl)ethyl, 2-(phenylsulfonyl)ethyl, vinyl, allyl, and p-nitrobenzyl carbonate. Examples of silyl ethers include trimethylsilyl, triethylsilyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, triisopropyl ether, and other trialkylsilyl ethers. Examples of alkyl ethers include methyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, trityl, t-butyl, and allyl ether, or derivatives thereof. Examples of alkoxyalkyl ethers include acetals, such as methoxymethyl, methylthiomethyl, (2-methoxyethoxy)methyl, benzoyloxymethyl, beta-(trimethylsilyl)ethoxymethyl, and tetrahydropyran-2-yl ether.Examples of arylalkyls include benzyl, p-methoxybenzyl (MPM), 3,4-dimethoxybenzyl, O-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, 2- and 4-picolyl ether.

[0029] Protected amines are well known in the art and include those described in detail in Greene (1999). Mono-protected amines further include, but are not limited to, aralkylamines, carbamates, allylamines, amides, etc. Examples of mono-protected amino moieties include t-butyloxycarbonylamino (-NHBOC), ethyloxycarbonylamino, methyloxycarbonylamino, trichloroethyloxycarbonylamino, allyloxycarbonylamino (-NHAlloc), benzyloxycarbonylamino (-NHCBZ), allylamino, benzylamino (-NHBn), fluorenylmethylcarbonyl (-NHFmoc), formamide, acetamide, chloroacetamide, dichloroacetamide, trichloroacetamide, phenylacetamide, trifluoroacetamide, benzamide, t-butyldiphenylsilyl, etc. Di-protected amines include amines substituted with two substituents independently selected from those described above as mono-protected amines, and further include cyclic imides such as phthalimide, maleimide, succinimide, etc. Also, di-protected amines include pyrrole, 2,2,5,5-tetramethyl-[1.2.5]azadisilolidine, etc., and azide.

[0030] ​Protected aldehydes are well known in the art and include those described in detail in Greene (1999). Protected aldehydes further include, but are not limited to, acyclic acetals, cyclic acetals, hydrazones, imines, and the like. Examples of such groups include dimethyl acetal, diethyl acetal, diisopropyl acetal, dibenzyl acetal, bis(2-nitrobenzyl) acetal, 1,3-dioxane, 1,3-dioxolane, semicarbazone, and derivatives thereof.

[0031] Protected carboxylic acids are well known in the art and include those described in detail in Greene (1999). Protected carboxylic acids further include, but are not limited to, optionally substituted C 1-6 aliphatic esters, optionally substituted aryl esters, silyl esters, activated esters, amides, hydrazides, and the like. Examples of such ester groups include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, benzyl, and phenyl esters, where each group is optionally substituted. Additional protected carboxylic acids include oxazoline and ortho esters.

[0032] Protected thiols are well known in the art and include those described in detail in Greene (1999). Protected thiols further include, but are not limited to, disulfides, thioethers, silyl thioethers, thioesters, thiocarbonates, and thiocarbamates. Examples of such groups include, but are not limited to, alkyl thioethers, benzyl and substituted benzyl thioethers, triphenylmethyl thioether, and trichloroethoxycarbonyl thioester.

[0033] Unless otherwise indicated, the structures shown in this specification are intended to include all isomeric forms (e.g., enantiomers, diastereomers, and geometric (or conformational)) in the structure, such as the R and S configurations at each chiral center, Z and E double bond isomers, and Z and E conformational isomers. Therefore, single stereochemical isomers of the compounds of the present invention, as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures, are within the scope of the present invention. Unless otherwise indicated, all tautomeric forms of the compounds of the present invention are within the scope of the present invention. In addition, unless otherwise indicated, the structures shown in this specification are intended to include compounds that differ only in the presence of one or more isotope-enriched atoms. For example, compounds having the structure of the present invention except that hydrogen is replaced by deuterium or tritium, or carbon is replaced by 13 C or 14 C-enriched carbon are within the scope of the present invention. Such compounds are useful, for example, as analytical tools or probes in biological assays, as in neutron scattering experiments.

[0034] As used herein, the expression "unit dosage form" refers to physically discrete units of the pharmaceutical formulation of the present invention appropriate for the subject to be treated. However, it should be understood that the total daily usage amount of the composition of the present invention is determined by the attending physician within the scope of reasonable medical judgment. The specific effective dosage level for any particular subject or organism will depend on a variety of factors, including the disorder being treated and the severity of the disorder, the activity of the specific active agent being used, the specific composition being used, the age, weight, general health, sex, and diet of the subject, the time of administration, the excretion rate of the specific active agent being used, the duration of the treatment, drugs and / or additional therapies used in combination with or simultaneously with the specific compound(s) being used, and similar factors well known in the medical arts.

[0035] The term "about" when referring to a measurable value such as an amount, a temporal period, etc., refers to a variation of ±20% of the specified value, or in some cases ±10%, or in some cases ±5%, or in some cases ±1%, or in some cases ±0.1% since the following variations are appropriate for carrying out the methods of the present disclosure.

[0036] 3. Description of Exemplary Embodiments 3.1 Active Pharmaceutical Ingredient In certain embodiments, the compounds of the invention are generally prepared according to Scheme I shown below. Scheme I

Chemical formula

[0037] In one aspect, the present invention provides a method for preparing a compound of formula I according to the steps shown in Scheme I above. In step S-1, ruthenium(III) chloride acid reacts with indazole to form the indazolium salt of trans-[tetrachlorobis(1H-indazole)ruthenium(III)] acid. This step (S-1) is well known in the art. See Keppler et. al. Inorganic Chemistry, 26, 1987. In step S-2, the indazolium salt is converted to the cesium salt of trans-[tetrachlorobis(1H-indazole)ruthenium(III)] of formula I-a by treatment with cesium chloride. Those skilled in the art recognize this as a salt exchange from the indazolium salt to the cesium salt. In step S-3, the cesium salt of formula I-a is converted to the sodium salt of trans-[tetrachlorobis(1H-indazole)ruthenium(III)] of formula I-b by treatment with sodium aluminum sulfate. Those skilled in the art recognize this as a salt exchange from the cesium salt to the sodium salt.

[0038] In certain embodiments, each of the above-described synthetic steps may be carried out sequentially, with isolation of each intermediate being carried out after each step. Alternatively, each of steps S-1, S-2, and S-3 shown in Scheme I above may be carried out in a manner in which isolation of the intermediate is not carried out.

[0039] One of ordinary skill in the art will recognize that steps S-1, S-2, and S-3 involve the preparation of a first indazolium salt in trans-[tetrachlorobis(1H-indazole)ruthenate(III)], followed by the preparation of the cesium salt, and then the preparation of the sodium salt. Further, U.S. Patent No. 8,362,266 describes the direct preparation of formula I-b from an indazolium salt. One aspect of the present invention involves the preparation of formula I-a as an intermediate in the synthesis of formula I-b. The cesium salt intermediate has been found to be preferred over existing methods because the purity and overall yield of the product can be significantly increased compared to existing methods. The inventors do not wish to be bound by any particular theory, but believe that the reason for this increase in yield and purity is due to the difficulty in isolating the indazolium salt of trans-[tetrachlorobis(1H-indazole)ruthenate(III)]. The inventors have found that for this material, since the filtered material retains residual water and hydrochloric acid, isolation as a solvent-free pure substance is very difficult. One proposed decomposition pathway for this material is shown in Scheme II below. Scheme II

Chemical formula

[0040] Scheme II shows compound A (mer, trans-[Ru IIIShows the preparation of [Cl3(Hind)2(H2O)]. The impurity compound A is also known in the literature as a water complex. The formation of compound A can be restricted by the elimination of water or by maintaining a clearly high concentration of chloride ions. For example, formula I-b is much more stable in a solution of sodium chloride or hydrochloric acid than in pure water. Those skilled in the art will recognize that maintaining the chloride ion concentration reduces the likelihood of the chlorine on the ruthenium complex being replaced by water. Furthermore, it has been found that the rate of aquation (or the preparation of compound A) increases significantly in basic solutions.

[0041] Since the primary decomposition product is a reactant that is promoted in an aquation reaction, specifically in a basic aqueous solution, it is considered preferable to avoid the reaction step involving the dissolution of the compound of formula I in water.

[0042] One embodiment of the present invention provides a method for preparing formula I-b by preparing trans-[tetrachlorobis(1H-indazole)ruthenate(III)] indazolium, isolating the material by filtration, and drying the material to 200% to 500% by mass of the theoretical yield for use in S-2. In other embodiments, the present invention provides a method for preparing formula I-b by preparing trans-[tetrachlorobis(1H-indazole)ruthenate(III)] indazolium, isolating the material by filtration, and drying the material to 245% to 425% by mass of the theoretical yield for use in S-2.

[0043] Another aspect of the present invention is to introduce step S-2 into the preparation of formula I-b. Step S-2 involves the preparation of the cesium intermediate of formula I-a. Surprisingly, the cesium intermediate can be isolated by precipitation and filtration, dried without inducing decomposition (as observed with indazolium salts), and the dried powder is stable under ambient conditions, so the cesium intermediate has been found to be an important step in the present invention. As described above, trans-[tetrachlorobis(1H-indazole)ruthenate(III)] indazolium is isolated by filtration as a substance that can best be described as a muddy substance. The stability of this compound is improved by the presence of hydrochloric acid (chloride ions). Washing the filtrate with a polar solvent (e.g., methanol) also leads to decomposition. Therefore, the best case is to prepare trans-[tetrachlorobis(1H-indazole)ruthenate(III)] indazolium, isolate it by filtration, and use it directly in S-2 without delay. S-2 consists of mixing trans-[tetrachlorobis(1H-indazole)ruthenate(III)] indazolium and cesium chloride in a suitable solvent. Suitable solvents can be alcohols having 1 to 5 carbon atoms, diols having 2 to 4 carbon atoms, water, ketones having 1 to 6 carbon atoms, cyclic ethers having 4 to 7 carbon atoms, amides having 1 to 4 carbon atoms, DMSO, sulfolane, esters having 4 to 6 carbon atoms, chlorinated hydrocarbons having 1 or 2 carbon atoms, liquid aromatic hydrocarbons, nitriles having 2 to 6 carbon atoms, or mixtures thereof.

[0044] In one embodiment of the present invention, S-2 consists of mixing trans-[tetrachlorobis(1H-indazole)ruthenate(III)] indazolium and cesium chloride in ethanol and methyl ethyl ketone to obtain Formula I-a. The cesium salt intermediate was collected by filtering the reaction mixture and washing with ethanol. In some embodiments, S-2 utilizes 1 to 10 equivalents of cesium chloride present in the reaction mixture. In other embodiments, S-2 utilizes 2 to 4 equivalents of cesium chloride present in the reaction mixture. In a preferred embodiment, the present invention provides a method for preparing Formula I-b, wherein 2.8 equivalents of cesium chloride are used in step S-2. The preferred solvent in S-2 is an ethanol-containing mixture, most preferably a methanol-methyl ethyl ketone (MEK) mixture. This is because the MEK mixture can form a crystalline MEK solvate of the cesium salt that aids in purification. The resulting MEK solvate in this case is then easily converted to a more stable hydrate form of the cesium salt by treatment with aqueous ethanol.

[0045] Another aspect of the present invention is step S-3 of converting a cesium salt intermediate (Formula I-a) to the desired sodium salt, Formula I-b. Previous methodologies for providing Formula I-b as described herein involve treating an aqueous solution of trans-[tetrachlorobis(1H-indazole)ruthenate(III)] with a sodium salt under basic conditions. As described above, the aqueous basic conditions lead to decomposition to Compound A. To address this problem, the inventors developed step S-3 of converting Formula I-a to Formula I-b by mixing sodium aluminum sulfate (NaAl(SO4)2). This salt exchange is carried out by mixing sodium aluminum sulfate and Formula I-a in water. The reaction is carried out at a high concentration such that the reaction mixture becomes heterogeneous. The driving force in the reaction is the differential solubility of sodium aluminum sulfate and cesium aluminum sulfate. Sodium aluminum sulfate is soluble in water and provides a source of sodium ions. Cesium aluminum sulfate is insoluble in water and precipitates from the reaction mixture. Thus, the cesium counterion is continuously removed from the reaction solution, and as a result, Formula I-b is formed. The insoluble cesium aluminum sulfate and Formula I-b are isolated by filtration. Formula I-b is dissolved in a suitable solvent, and cesium aluminum sulfate is removed by filtration. Suitable solvents include low molecular weight alcohols (having 1 to 5 carbon atoms), ketones having 3 to 6 carbon atoms, nitriles having 2 to 5 carbon atoms, esters having 3 to 6 carbon atoms, amides having 1 to 4 carbon atoms, diols having 1 to 4 carbon atoms, DMSO, sulfolane, water, or combinations thereof. The most preferred solvent for solid extraction is acetonitrile. Next, Formula I-b is precipitated using a suitable anti-solvent and recovered by filtration. Suitable anti-solvents include ethers having 3 to 8 carbon atoms, cyclic, acyclic, or aromatic hydrocarbons having 5 to 8 carbon atoms, chlorinated hydrocarbons having 1 to 4 carbon atoms, benzotrifluoride, chlorobenzene, methyl carbonate. The most preferred anti-solvent is methyl tert-butyl ether (MTBE).

[0046] In some embodiments, the present invention provides a method for preparing formula I-b, wherein the concentration of sodium aluminum sulfate in step S-3 is 0.5 M to 1.65 M. In a preferred embodiment, the present invention provides a method for preparing formula I-b, wherein the concentration of sodium aluminum sulfate in step S-3 is 1.1 M.

[0047] In some embodiments, the present invention provides a method for preparing formula I-b, wherein the reaction temperature in step S-3 is -5 °C to 50 °C. In a preferred embodiment, the present invention provides a method for preparing formula I-b, wherein the reaction temperature in step S-3 is 20 °C to 25 °C.

[0048] In some embodiments, the present invention provides a method for preparing formula I-b, wherein the reaction time in step S-3 is 12 hours to 168 hours. In a preferred embodiment, the present invention provides a method for preparing formula I-b, wherein the reaction time in step S-3 is 30 hours.

[0049] Another aspect of the present invention is a purification step in which residual cesium is removed from formula I-b. This process involves stirring formula I-b with methanol in the presence of 4 Å molecular sieves and then precipitating with MTBE. Without wishing to be bound by any particular theory, it is believed that cesium atoms have an affinity for the 4 Å pores present within the molecular sieves. Furthermore, it has been found that by stirring and washing with an MTBE solution saturated with water, trace amounts of solvent impurities can be removed from the desired product. By using this final purification step, the highest purity of formula I-b can be obtained.

[0050] Multiple techniques were required for the characterization of ruthenium containing the target compound. Nuclear magnetic resonance spectroscopy of ruthenium compounds is difficult due to the 5 / 2 nuclear spin state, so alternative characterization methods including HPLC and x-ray diffraction (crystal analysis) were used. To fully characterize the purity of IT-139, the inventors intentionally prepared a plurality of compounds considered to be impurities in the final composition of IT-139, namely compounds A, C, and D. The identities of impurities A, C, and D were confirmed by x-ray diffraction. Compound B is an unstable complex considered to be an intermediate in the formation of compound C. The structures of the impurities are as follows. [Chemical formula]

[0051] Once the impurity compounds were prepared and identified, the retention times of these impurities were analyzed by HPLC so that the identity and percentage of the impurities could be quickly quantified by HPLC analysis. During this process, the inventors observed that compound A, which is a water complex, gave rise to multiple peaks on HPLC and that the chromatographic profile changed as a function of time. It was found that the water complex reacted with acetonitrile in the mobile phase to form compound B, which is an acetonitrile adduct, and subsequently this adduct reacted to form compound C, which is a covalent derivative with acetonitrile (see Inorganic Chemistry, 2008, v47, p6513 - 6523). This reaction is shown in Scheme III below. It was found that it forms compound C, which is a covalent derivative with acetonitrile (see Inorganic Chemistry, 2008, v47, p6513 - 6523). This reaction is shown in Scheme III below. [Chemical formula]

[0052] The relative retention times for each compound are listed in the table below (Table 1). [Table 1]

[0053] In some embodiments, the relative retention time (RRT) described in Table 1 can be defined by a range. For example, the RRT of Compound A can be 1.09 + / - 0.02, the RRT of Compound B can be 1.28 + / - 0.02, the RRT of Compound C can be 1.06 + / - 0.03, and the RRT of Compound D can be 1.59 + / - 0.03.

[0054] Since the water complex (Compound A) rapidly forms Compounds B and C in the mobile phase in HPLC analysis, the amount of Compound A in the sample passed for HPLC analysis is determined to be the sum of the peak areas corresponding to Compounds A, B, and C. One advantage of the synthetic methodology of the present invention over other synthetic methodologies is the high purity level that can be achieved by the present invention. The previous methodologies described above result in a final product (drug substance) containing 4 - 8% of Compound A as impurities. In contrast, in the present invention, less than 2% of Compound A can be easily achieved. One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] and Compound A, wherein the composition has 2.0 wt% or less of Compound A present therein. One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] and Compound A, wherein the composition has 1.0 wt% or less of Compound A present therein. One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] and Compound A, wherein the composition has 1.5 wt% or less of Compound A present therein. One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] and Compound A, wherein the composition has 0.5 wt% or less of Compound A present therein. One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] and Compound A, wherein the composition has 3.0 wt% or less of Compound A present therein.

[0055] Another advantage of the present invention over previous synthetic methodologies is the reduction in the amount of impurities to be purified. The previous synthetic methodologies described above (see, for example, U.S. Patent No. 8,362,266) were often analyzed by an HPLC method (e.g., HPLC method #3 below) that does not decompose impurities (Compound A, Compound B, and Compound C) from the active pharmaceutical ingredient (Formula I-b). Figure 4 shows the active pharmaceutical ingredient prepared using other synthetic methodologies analyzed by HPLC method #3, while Figure 5 shows the same active pharmaceutical ingredient analyzed by an analytical method (HPLC method #2 below) that decomposes Formula I-b from impurity compounds A, B, and C. As a result, a purity of about 99.5% was reported for the active pharmaceutical ingredient synthesized using other methodologies (analysis by HPLC method #3). However, it was shown that the same material analyzed by HPLC method #2 actually contained approximately 76.4% of Formula I-b, approximately 7.3% of Compound A, approximately 11.0% of Compound B, and approximately 0.36% of Compound C. The present invention provides a composition containing Formula I-b with a purity of about 99.9% in an analysis using HPLC method #3. The present invention provides a composition containing about 96.3% of Formula I-b, about 1.1% of Compound A, about 1.7% of Compound B, and about 0.2% of Compound C. The HPLC data is reproduced in the table below (Table 2).

Table 2

[0056] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], Ru III Cl3(Hind)2(H2O), Ru III Cl3(Hind)2(CH3CN), and Ru III Cl3(Hind)(HN=C(Me)ind).

[0057] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], Ru III Cl3(Hind)2(H2O), Ru III Cl3(Hind)2(CH3CN), and RuIII Provide a composition comprising Cl3(Hind)(HN=C(Me)ind) and cesium.

[0058] One embodiment of the present invention is sodium trans - [tetrachlorobis(1H - indazole)ruthenate(III)], Ru III Cl3(Hind)2(H2O), Ru III Cl3(Hind)2(CH3CN), Ru III A composition comprising Cl3(Hind)(HN=C(Me)ind) and cesium, wherein the sodium trans - [tetrachlorobis(1H - indazole)ruthenate(III)] is at least about 95.5 weight percentage of the composition, the Ru III Cl3(Hind)2(H2O) is at most about 1.0 weight percentage of the composition, the Ru III Cl3(Hind)2(CH3CN) is at most about 2.5 weight percentage of the composition, the Ru III Cl3(Hind)(HN=C(Me)ind) is at most about 2.0 weight percentage of the composition, cesium is at most about 0.5 weight percentage of the composition, Provide a composition.

[0059] One embodiment of the present invention is a composition comprising sodium trans - [tetrachlorobis(1H - indazole)ruthenate(III)], cesium, and optionally Ru III Cl3(Hind)2(H2O), Ru III Cl3(Hind)2(CH3CN), Ru III Cl3(Hind)(HN=C(Me)ind), wherein the sodium trans - [tetrachlorobis(1H - indazole)ruthenate(III)] is from about 95.5 to about 99.9 weight percentage of the composition, the Ru IIICl3(Hind)2(H2O) is from about 0 to about 1.0 weight percentage of the composition, the Ru III Cl3(Hind)2(CH3CN) is from about 0 to about 2.5 weight percentage of the composition, the Ru III Cl3(Hind)(HN=C(Me)ind) is from about 0 to about 2.0 weight percentage of the composition, cesium is from about 0 to about 0.5 weight percentage of the composition, providing a composition.

[0060] One embodiment of the present invention is a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], Ru III Cl3(Hind)2(H2O), Ru III Cl3(Hind)2(CH3CN), Ru III Cl3(Hind)(HN=C(Me)ind), and cesium, wherein the sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] is from about 95.5 to about 99.9 weight percentage of the composition, the Ru III Cl3(Hind)2(H2O) is from about 0.001 to about 1.0 weight percentage of the composition, the Ru III Cl3(Hind)2(CH3CN) is from about 0.001 to about 2.5 weight percentage of the composition, the Ru III Cl3(Hind)(HN=C(Me)ind) is from about 0.001 to about 2.0 weight percentage of the composition, cesium is from about 0.0001 to about 0.5 weight percentage of the composition, providing a composition.

[0061] One embodiment of the present invention is a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], Ru IIICl3(Hind)2(H2O), and Ru III Cl3(Hind)2(CH3CN), and Ru III Cl3(Hind)(HN=C(Me)ind), and cesium, a composition comprising the trans-[tetrachlorobis(1H-indazole)ruthenate(III)] sodium is from about 95.5 to about 99.9 weight percentage of the composition, the Ru III Cl3(Hind)2(H2O) is from about 0.001 to about 0.75 weight percentage of the composition, the Ru III Cl3(Hind)2(CH3CN) is from about 0.001 to about 1.5 weight percentage of the composition, the Ru III Cl3(Hind)(HN=C(Me)ind) is from about 0.001 to about 1.25 weight percentage of the composition, cesium is from about 0.0001 to about 0.25 weight percentage of the composition, to provide a composition.

[0062] One embodiment of the present invention is a composition comprising trans-[tetrachlorobis(1H-indazole)ruthenate(III)] sodium, Ru III Cl3(Hind)2(H2O), Ru III Cl3(Hind)2(CH3CN), Ru III Cl3(Hind)(HN=C(Me)ind), and cesium, a composition comprising the trans-[tetrachlorobis(1H-indazole)ruthenate(III)] sodium is from about 95.5 to about 99.9 weight percentage of the composition, the Ru III Cl3(Hind)2(H2O) is from about 0.001 to about 0.5 weight percentage of the composition, the Ru III Cl3(Hind)2(CH3CN) is from about 0.001 to about 0.5 weight percentage of the composition, the Ru IIICl3(Hind)(HN=C(Me)ind) is from about 0.001 to about 0.5 weight percentage of the composition, cesium is from about 0.0001 to about 0.01 weight percentage of the composition, providing a composition.

[0063] 3.2 Drugs An additional embodiment of the present invention provides a method for preparing a drug containing the sodium salt of trans - [tetrachlorobis(1H - indazole)ruthenate(III)] (i.e., IT - 139).

[0064] One aspect of the present invention provides a method for preparing a sterile lyophilized drug containing sodium trans - [tetrachlorobis(1H - indazole)ruthenate(III)]. This formulation is considered suitable for administration to patients. The formulation is composed of sodium trans - [tetrachlorobis(1H - indazole)ruthenate(III)], a pH buffer, and a cryoprotectant. General methods for providing the aforementioned formulation include the steps of preparing an aqueous buffer solution, preparing an aqueous cryoprotective substance solution, dissolving sodium trans - [tetrachlorobis(1H - indazole)ruthenate(III)] in the buffer solution, adding the cryoprotective substance solution, sterile filtration (e.g., aseptic filtration), filling vials under sterile conditions, and lyophilizing under sterile conditions. Suitable buffers include, but are not limited to, citric acid, TRIS, acetic acid, EDTA, HEPES, tricine, and imidazole. The use of phosphate buffer is possible but not preferred. A preferred embodiment of the present invention is the use of a citric acid / sodium citrate buffer. Suitable cryoprotectants include, but are not limited to, sugars, monosaccharides, disaccharides, polysaccharides, polyalcohols, mannitol, sorbi tol, sucrose, trehalose, dextran, and dextrose. A preferred embodiment of the present invention is the use of mannitol as a cryoprotectant.

[0065] As described above in this specification, sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] can decompose in water to form Compound A (Scheme II). Those skilled in the art will recognize that the limitations of this decomposition reaction will be advantageous in obtaining the highest purity product. It has been found that cooling sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] during the formulation process greatly reduces the amount of Compound A present in the lyophilized product. In one aspect of the present invention, the sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] solution is cooled to 4 °C during the formulation process. In another aspect of the present invention, the sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] solution is cooled to 2 - 8 °C during the formulation process. In another aspect of the present invention, the sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] solution is cooled to 2 - 15 °C during the formulation process.

[0066] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], a suitable buffer, and mannitol. In some embodiments, the suitable buffer comprises a citrate buffer. For example, in some embodiments, the citrate buffer comprises sodium citrate and citric acid.

[0067] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], sodium citrate, citric acid, and mannitol.

[0068] One embodiment of the present invention provides a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], sodium citrate, citric acid, mannitol, and mer, trans-[Ru III Cl3(Hind)2(H2O)].

[0069] One embodiment of the present invention is sodium [trans - tetrachlorobis(1H - indazole)ruthenate(III)], sodium citrate, citric acid, mannitol, mer, trans - [Ru III Cl3(Hind)2(H2O)], and a cesium salt.

[0070] One embodiment of the present invention provides a composition comprising sodium [trans - tetrachlorobis(1H - indazole)ruthenate(III)], sodium citrate, citric acid, and mannitol, wherein the sodium [trans - tetrachlorobis(1H - indazole)ruthenate(III)] is amorphous.

[0071] One embodiment of the present invention is a composition comprising sodium [trans - tetrachlorobis(1H - indazole)ruthenate(III)], sodium citrate, citric acid, mannitol, mer, trans - [Ru III Cl3(Hind)2(H2O)], wherein the sodium [trans - tetrachlorobis(1H - indazole)ruthenate(III)] is amorphous.

[0072] One embodiment of the present invention is a composition comprising sodium [trans - tetrachlorobis(1H - indazole)ruthenate(III)], sodium citrate, citric acid, mannitol, mer, trans - [Ru III Cl3(Hind)2(H2O)] and a cesium salt wherein the sodium [trans - tetrachlorobis(1H - indazole)ruthenate(III)] is amorphous.

[0073] One embodiment of the present invention is a composition comprising sodium [trans - tetrachlorobis(1H - indazole)ruthenate(III)], sodium citrate, citric acid, mannitol, mer, trans - [Ru III Cl3(Hind)2(H2O)] and a cesium salt, mer, trans-[Ru III Cl3(Hind)2(H2O)] is from about 0.01 to about 0.4 weight percent of the composition, cesium is from about 0.00001 to about 0.01 weight percent of the composition, and provides a composition.

[0074] One embodiment of the present invention is a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], sodium citrate, citric acid, mannitol, mer, trans-[Ru III Cl3(Hind)2(H2O)], and a cesium salt, wherein mer, trans-[Ru III Cl3(Hind)2(H2O)] is from about 0.01 to about 0.4 weight percent of the composition, cesium is from about 0.00001 to about 0.01 weight percent of the composition, and provides a composition.

[0075] One embodiment of the present invention is a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], sodium citrate, citric acid, mannitol, mer, trans-[Ru III Cl3(Hind)2(H2O)], and a cesium salt, wherein mer, trans-[Ru III Cl3(Hind)2(H2O)] is from about 0.01 to about 0.2 weight percent of the composition, cesium is from about 0.00001 to about 0.01 weight percent of the composition, and provides a composition.

[0076] One embodiment of the present invention is a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], mer, trans-[Ru III Cl3(Hind)2(H2O)], and a cesium salt, wherein mer, trans-[RuIII [RuCl3(Hind)2(H2O)] is from about 0.01 to about 0.40 weight percent of the composition, cesium is from about 0.00001 to about 0.01 weight percent of the composition, providing a composition.

[0077] One embodiment of the present invention is a composition comprising sodium trans - [tetrachlorobis(1H - indazole)ruthenate(III)], mer,trans - [Ru III Cl3(Hind)2(H2O)], and a cesium salt, wherein the composition is a lyophilized powder, mer,trans - [Ru III Cl3(Hind)2(H2O)] is from about 0.01 to about 0.40 weight percent of the composition, cesium is from about 0.00001 to about 0.01 weight percent of the composition, providing a composition.

[0078] One embodiment of the present invention is a composition comprising sodium trans - [tetrachlorobis(1H - indazole)ruthenate(III)], sodium citrate, citric acid, mannitol, mer,trans - [Ru III Cl3(Hind)2(H2O)], and a cesium salt wherein the composition is a lyophilized powder, mer,trans - [Ru mer,trans - [Ru III Cl3(Hind)2(H2O)] is from about 0.01 to about 0.3 weight percent of the composition, cesium is from about 0.00001 to about 0.1 weight percent of the composition, providing a composition.

[0079] One embodiment of the present invention is a composition comprising sodium trans - [tetrachlorobis(1H - indazole)ruthenate(III)], sodium citrate, citric acid, mannitol, mer,trans - [Ru IIIA composition comprising [RuCl3(Hind)2(H2O)] and a cesium salt, mer, trans-[Ru III [RuCl3(Hind)2(H2O)] is from about 0.01 to about 0.3 weight percent of the composition, cesium is from about 0.00001 to about 0.1 weight percent of the composition, provides a composition.

[0080] One embodiment of the present invention is a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], sodium citrate, citric acid, mannitol, mer, trans-[Ru III [RuCl3(Hind)2(H2O)] and a cesium salt, the composition is a lyophilized powder, sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] is from about 11.5 to about 14.0 weight percent of the composition, citric acid is from about 43.9 to about 53.7 weight percent of the composition, sodium citrate is from about 25.7 to about 23.1 weight percent of the composition, mannitol is from about 11.5 to about 14.0 weight percent of the composition, mer, trans-[Ru III [RuCl3(Hind)2(H2O)] is from about 0.01 and about 0.3 weight percent of the composition, cesium is from about 0.00001 to about 0.1 weight percent of the composition, provides a composition.

[0081] One embodiment of the present invention is a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], sodium citrate, citric acid, mannitol, mer, trans-[Ru III [RuCl3(Hind)2(H2O)] and a cesium salt, the composition is a lyophilized powder, Sodium trans - [tetrachlorobis(1H - indazole)ruthenate(III)] is from about 10.2 to about 15.3 weight percent of the composition, citric acid is from about 39.0 to about 58.5 weight percent of the composition, sodium citrate is from about 20.5 to about 30.8 weight percent of the composition, mannitol is from about 10.2 to about 15.3 weight percent of the composition, mer, trans - [Ru III Cl3(Hind)2(H2O)] is from about 0.01 and about 0.3 weight percent of the composition, cesium is from about 0.00001 to about 0.1 weight percent of the composition, and provides a composition.

[0082] One embodiment of the present invention is a composition comprising sodium trans - [tetrachlorobis(1H - indazole)ruthenate(III)], sodium citrate, citric acid, mannitol, mer, trans - [Ru III Cl3(Hind)2(H2O)], and a cesium salt, wherein the composition is a lyophilized powder, sodium trans - [tetrachlorobis(1H - indazole)ruthenate(III)] is from about 10.2 to about 15.3 weight percent of the composition, mer, trans - [Ru III Cl3(Hind)2(H2O)] is from about 0.01 and about 0.3 weight percent of the composition, cesium is from about 0.00001 to about 0.1 weight percent of the composition, and provides a composition.

[0083] One embodiment of the present invention is a composition comprising sodium trans - [tetrachlorobis(1H - indazole)ruthenate(III)], mannitol, citric acid, and sodium citrate, Sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] is about 49.86 weight percent of the composition, mannitol is about 49.86 weight percent of the composition, citric acid is about 0.187 weight percent of the composition, sodium citrate is about 0.093 weight percentage of the composition, providing a composition. In some such embodiments, the composition is a lyophilized powder.

[0084] One embodiment of the present invention is a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], mannitol, citric acid, and sodium citrate, wherein sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] is about 40 to about 60 weight percent of the composition, mannitol is about 40 to about 60 weight percent of the composition, citric acid is about 0.01 to about 0.5 weight percent of the composition, and sodium citrate is about 0.001 to about 0.25 weight percentage of the composition, providing a composition. In some such embodiments, the composition is a lyophilized powder.

[0085] One embodiment of the present invention is a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], mannitol, citric acid, and sodium citrate, wherein sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] is about 30 to about 70 weight percent of the composition, mannitol is about 30 to about 70 weight percent of the composition, citric acid is about 0.001 to about 1 weight percent of the composition, Sodium citrate is from about 0.0001 to about 1 weight percentage of the composition, and provides a composition. In some such embodiments, the composition is a lyophilized powder.

[0086] One embodiment of the present invention is a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], mannitol, citric acid, sodium citrate, and Ru III Cl3(Hind)2(H2O), wherein sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] is about 49.86 weight percent of the composition, mannitol is about 49.86 weight percent of the composition, citric acid is about 0.187 weight percent of the composition, sodium citrate is about 0.093 weight percentage of the composition, Ru III Cl3(Hind)2(H2O) is 0.5 weight percent or less of the composition, and provides a composition. In some such embodiments, the composition is a lyophilized powder.

[0087] One embodiment of the present invention is a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], mannitol, citric acid, sodium citrate, and Ru III Cl3(Hind)2(H2O), wherein sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] is from about 40 to about 60 weight percent of the composition, mannitol is from about 40 to about 60 weight percent of the composition, citric acid is from about 0.01 to about 0.5 weight percent of the composition, sodium citrate is from about 0.001 to about 0.25 weight percentage of the composition, RuIII Cl3(Hind)2(H2O) is from about 0 to about 0.5 weight percentage of the composition, A composition is provided. In some such embodiments, the composition is a lyophilized powder.

[0088] One embodiment of the present invention is a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], mannitol, citric acid, sodium citrate, and Ru III Cl3(Hind)2(H2O), and cesium, wherein sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] is from about 30 to about 70 weight percentage of the composition, mannitol is from about 30 to about 70 weight percentage of the composition, citric acid is from about 0.001 to about 1 weight percentage of the composition, sodium citrate is from about 0.0001 to about 1 weight percentage of the composition, Ru III Cl3(Hind)2(H2O) is 0.5 weight percentage or less of the composition, cesium is 0.25 weight percentage or less of the composition, A composition is provided. In some such embodiments, the composition is a lyophilized powder.

[0089] One embodiment of the present invention is a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], mannitol, citric acid, sodium citrate, and cesium, wherein sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] is about 49.61 weight percentage of the composition, mannitol is about 49.86 weight percentage of the composition, citric acid is about 0.187 weight percentage of the composition, Sodium citrate is about 0.093 weight percentage of the composition, cesium is about 0.25 weight percentage of the composition, providing a composition. In some such embodiments, the composition is a lyophilized powder.

[0090] One embodiment of the present invention is a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], mannitol, citric acid, sodium citrate, and cesium, sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] is about 40 to about 60 weight percent of the composition, mannitol is about 40 to about 60 weight percent of the composition, citric acid is about 0.01 to about 0.5 weight percent of the composition, sodium citrate is about 0.001 to about 0.25 weight percentage of the composition, cesium is about 0.1 to about 0.5 weight percentage of the composition, providing a composition. In some such embodiments, the composition is a lyophilized powder.

[0091] One embodiment of the present invention is a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], mannitol, citric acid, sodium citrate, and cesium, sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] is about 30 to about 70 weight percent of the composition, mannitol is about 30 to about 70 weight percent of the composition, citric acid is about 0.001 to about 1 weight percent of the composition, sodium citrate is about 0.0001 to about 1 weight percentage of the composition, cesium is about 0.01 to about 1 weight percentage of the composition, A composition is provided. In some such embodiments, the composition is a lyophilized powder.

[0092] One embodiment of the present invention is a composition comprising sodium trans - [tetrachlorobis(1H - indazole)ruthenate(III)], mannitol, citric acid, sodium citrate, Ru III Cl3(Hind)2(H2O), Ru III Cl3(Hind)2(CH3CN), Ru III Cl3(Hind)(HN = C(Me)ind), and cesium, wherein sodium trans - [tetrachlorobis(1H - indazole)ruthenate(III)] is about 46.61 weight percent of the composition, mannitol is about 49.86 weight percent of the composition, citric acid is about 0.187 weight percent of the composition, sodium citrate is about 0.093 weight percentage of the composition, Ru III Cl3(Hind)2(H2O) is 0.5 weight percentage or less of the composition, Ru III Cl3(Hind)2(CH3CN) is 1.25 weight percentage or less of the composition, Ru III Cl3(Hind)(HN = C(Me)ind) is 1.0 weight percentage or less of the composition, cesium is 0.25 weight percentage or less of the composition. A composition is provided. In some such embodiments, the composition is a lyophilized powder.

[0093] One embodiment of the present invention is a composition comprising sodium trans - [tetrachlorobis(1H - indazole)ruthenate(III)], mannitol, citric acid, sodium citrate, Ru III Cl3(Hind)2(H2O), Ru IIICl3(Hind)2(CH3CN) and Ru III A composition comprising Cl3(Hind)(HN=C(Me)ind) and cesium, where sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] is about 46.61 weight percent of the composition, where mannitol is about 49.86 weight percent of the composition, where citric acid is about 0.187 weight percent of the composition, where sodium citrate is about 0.093 weight percentage of the composition, Ru III where RuCl3(Hind)2(H2O) is 0.5 weight percentage or less of the composition, Ru III where RuCl3(Hind)2(CH3CN) is 1.25 weight percentage or less of the composition, Ru III where RuCl3(Hind)(HN=C(Me)ind) is 1.0 weight percentage or less of the composition, and cesium is 0.25 weight percentage or less of the composition. Provided is a composition. In some such embodiments, the composition is a lyophilized powder.

[0094] One embodiment of the present invention is a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], mannitol, citric acid, sodium citrate, Ru III RuCl3(Hind)2(H2O), Ru III RuCl3(Hind)2(CH3CN), Ru III RuCl3(Hind)(HN=C(Me)ind), and cesium, where sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] is about 40 to about 60 weight percent of the composition, where mannitol is about 40 to about 60 weight percent of the composition, Citric acid is from about 0.01 to about 0.5 weight percent of the composition, sodium citrate is from about 0.001 to about 0.25 weight percentage of the composition, Ru III Cl3(Hind)2(H2O) is at most about 0.5 weight percentage of the composition, Ru III Cl3(Hind)2(CH3CN) is at most about 1.25 weight percentage of the composition, Ru III Cl3(Hind)(HN=C(Me)ind) is at most about 1.0 weight percentage of the composition, cesium is at most 0.25 percentage of the composition, and provides a composition. In some such embodiments, the composition is a lyophilized powder.

[0095] One embodiment of the present invention is a composition comprising sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], mannitol, citric acid, sodium citrate, Ru III Cl3(Hind)2(H2O), Ru III Cl3(Hind)2(CH3CN), Ru III Cl3(Hind)(HN=C(Me)ind), and cesium, wherein sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] is from about 30 to about 70 weight percent of the composition, mannitol is from about 30 to about 70 weight percent of the composition, citric acid is from about 0.001 to about 1 weight percent of the composition, sodium citrate is from about 0.0001 to about 1 weight percentage of the composition, Ru III Cl3(Hind)2(H2O) is at most about 0.5 weight percentage of the composition, Ru IIICl3(Hind)2(CH3CN) is at most about 1.25 weight percentage of the composition, Ru III Cl3(Hind)(HN=C(Me)ind) is at most about 1.0 weight percentage of the composition, Cesium is at most 0.25 percentage of the composition, A composition is provided. In some such embodiments, the composition is a lyophilized powder.

[0096] One embodiment of the present invention is a composition comprising sodium trans - [tetrachlorobis(1H - indazole)ruthenate(III)], mannitol, citric acid, sodium citrate, Ru III Cl3(Hind)2(H2O), Ru III Cl3(Hind)2(CH3CN), Ru III Cl3(Hind)(HN=C(Me)ind), and cesium, wherein sodium trans - [tetrachlorobis(1H - indazole)ruthenate(III)] is about 20 to about 80 weight percent of the composition, mannitol is about 20 to about 80 weight percent of the composition, citric acid is about 0.0001 to about 5 weight percent of the composition, sodium citrate is about 0.00001 to about 5 weight percentage of the composition, Ru III Cl3(Hind)2(H2O) is at most about 0.5 weight percentage of the composition, Ru III Cl3(Hind)2(CH3CN) is at most about 1.25 weight percentage of the composition, Ru III Cl3(Hind)(HN=C(Me)ind) is at most about 1.0 weight percentage of the composition, cesium is at most 0.25 percentage of the composition, A composition is provided. In some such embodiments, the composition is a lyophilized powder.

[0097] 3.3 Unit Dosage Forms In some embodiments, the present invention provides unit dosage forms comprising the formulations or compositions described herein. As used herein, the expression "unit dosage form" refers to physically discrete units of a formulation provided as appropriate for the subject to be treated, it being understood that the total daily usage of the formulation provided will be determined within the scope of sound medical judgment by the attending physician. It should be understood that the specific effective dosage level for any particular subject or organism will depend upon a variety of factors including the disorder being treated and the severity of the disorder, the activity of the specific active agent being used, the specific formulation being used, the age, weight, general health, sex, and diet of the subject, the time of administration, the excretion rate of the specific active agent being used, the duration of the treatment, drugs and / or additional therapies used in combination with or simultaneously with the specific compound(s) being used, and like factors well known in the medical arts.

[0098] The compositions of the present invention can be provided as unit dosage forms. In some embodiments, a vial containing sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], mannitol, citric acid, and sodium citrate is a unit dosage form.

[0099] In some embodiments, a vial containing sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], mannitol, citric acid, sodium citrate, and cesium of the present invention is a unit dosage form.

[0100] In some embodiments, sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], mannitol, citric acid, sodium citrate, and Ru III Cl3(Hind)2(H2O) and Ru III Cl3(Hin d)2(CH3CN) and Ru IIIThe vial containing Cl3(Hind)(HN=C(Me)ind) and cesium is a unit dosage form.

[0101] Even further included in the present invention are pharmaceutical packs and / or kits containing the compositions described herein, or unit dosage forms comprising the compositions provided and a container (e.g., a foil or plastic package, or other suitable container). Optionally, instructions for use are additionally provided in such kits.

[0102] In some embodiments, the present invention may be provided as a unit dosage form. In fact, the vial containing sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)], mannitol, citric acid, and sodium citrate is a unit dosage form as shown in Table 3. [Table 3]

[0103] In some embodiments, the pharmaceutical components described in Table 3 further contain cesium, At this time, cesium is 0.25 weight percentage or less of the composition.

[0104] In some embodiments, the pharmaceutical components described in Table 3 contain cesium, Ru III Cl3(Hind)2(H2O), Ru III Cl3(Hind)2(CH3CN), Ru III Cl3(Hind)(HN=C(Me)ind) further, At this time, cesium is about 0.25 weight percentage or less of the composition, Ru III Cl3(Hind)2(H2O) is about 0.5 weight percentage or less of the composition, Ru III Cl3(Hind)2(CH3CN) is about 1.25 weight percentage or less of the composition, Ru III Cl3(Hind)(HN=C(Me)ind) is at most about 1.0 weight percentage of the composition.

[0105] In some embodiments, the pharmaceutical composition is selected from those in Table 4.

Table 4

[0106] In some embodiments, the pharmaceutical components described in Table 4 further include cesium, At this time, cesium is at most 0.25 weight percentage of the composition.

[0107] In some embodiments, the pharmaceutical components described in Table 4 include cesium, Ru III Cl3(Hind)2(H2O), Ru III Cl3(Hind)2(CH3CN), and Ru III Cl3(Hind)(HN=C(Me)ind), At this time, cesium is at most about 0.25 weight percentage of the composition, Ru III Cl3(Hind)2(H2O) is at most about 0.5 weight percentage of the composition, Ru III Cl3(Hind)2(CH3CN) is at most about 1.25 weight percentage of the composition, Ru III Cl3(Hind)(HN=C(Me)ind) is at most about 1.0 weight percentage of the composition.

[0108] In some embodiments, the present invention can be provided in unit dosage form. In fact, a vial containing sodium [tetrachlorobis(1H-indazole)ruthenate(III)], mannitol, citric acid, and sodium citrate is a unit dosage form shown in Table 5.

Table 5

[0109] In some embodiments, the pharmaceutical components described in Table 5 further include cesium, At this time, cesium is 0.25 weight percentage or less of the composition.

[0110] In some embodiments, the pharmaceutical components described in Table 5 include cesium, Ru III Cl3(Hind)2(H2O), Ru III Cl3(Hind)2(CH3CN), and Ru III Cl3(Hind)(HN=C(Me)ind), At this time, cesium is about 0.25 weight percentage or less of the composition, Ru III Cl3(Hind)2(H2O) is about 0.5 weight percentage or less of the composition, Ru III Cl3(Hind)2(CH3CN) is about 1.25 weight percentage or less of the composition, Ru III Cl3(Hind)(HN=C(Me)ind) is about 1.0 weight percentage or less of the composition.

[0111] In some embodiments, the pharmaceutical composition is selected from those in Table 6.

Table 6

[0112] In some embodiments, the pharmaceutical components described in Table 6 further include cesium, At this time, cesium is 0.25 weight percentage or less of the composition.

[0113] In some embodiments, the pharmaceutical components listed in Table 6 are cesium and Ru III Cl3(Hind)2(H2O) and Ru III Cl3(Hind)2(CH3CN) and Ru III Cl3(Hind)(HN=C(Me)ind), and at this time, cesium is 0.25 weight percentage or less of the composition, Ru III Cl3(Hind)2(H2O) is 0.5 weight percentage or less of the composition, Ru III Cl3(Hind)2(CH3CN) is 1.25 weight percentage or less of the composition, Ru III Cl3(Hind)(HN=C(Me)ind) is about 1.0 weight percentage or less of the composition.

[0114] In some embodiments, the pharmaceutical components are as described in any of Tables 3-6 and further contain cesium. In some embodiments, cesium is present in an amount of about 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.010, 0.015, 0.020, 0.025, 0.030, 0.035, 0.040, 0.045, 0.050, 0.055, 0.060, 0.065, 0.070, 0.075, 0.080, 0.085, 0.090, 0.095, 0.10, 0.15, 0.20, 0.25, 0.30, 0.35, 0.40, 0.45, 0.50, 0.55, 0.60, 0.65, 0.70, 0.75, 0.80, 0.85, 0.90, 0.95, or 1.0 weight percentage of the composition.

[0115] 3.4 Treatment methods In some embodiments, the present invention provides a method for treating cancer in a subject in need of cancer treatment, the method comprising administering to the subject the composition of IT-139 provided above and described herein. In some such embodiments, the subject is a human patient.

[0116] In some embodiments, the present invention provides a method for treating cancer in a subject in need of cancer treatment, the method comprising administering the composition of IT-139 provided above and described herein in combination with a chemotherapeutic agent.

[0117] In some embodiments, the present invention provides a method for treating cancer in a subject in need of cancer treatment, the method comprising administering the composition of IT-139 provided above and described herein in combination with a cancer immunotherapeutic agent.

[0118] According to another embodiment, the present invention relates to a method for treating cancer selected from breast, ovarian, cervical, prostate, testicular, urogenital, esophageal, laryngeal, glioblastoma, neuroblastoma, gastric, skin, keratoacanthoma, lung, epidermoid carcinoma, large cell carcinoma, small cell carcinoma, lung adenocarcinoma, bone, colon, adenoma, pancreas, adenocarcinoma, thyroid, follicular carcinoma, undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma, sarcoma, bladder cancer, liver and biliary tract, kidney cancer, myeloid disorders, lymphatic disorders, Hodgkin, hairy cell, buccal cavity and pharynx (oral cavity), lip, tongue, mouth, pharynx, small intestine, colorectal, large intestine, rectum, brain and central nervous system, and leukemia, the method comprising administering IT-139 or a pharmaceutically acceptable composition thereof.

[0119] According to another embodiment, the present invention relates to a method for treating cancer selected from breast, ovary, cervix, prostate, testis, urogenital tract, esophagus, larynx, glioblastoma, neuroblastoma, stomach, skin, keratoacanthoma, lung, epidermoid carcinoma, large cell carcinoma, small cell carcinoma, lung adenocarcinoma, bone, colon, adenoma, pancreas, adenocarcinoma, thyroid, follicular carcinoma, undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma, sarcoma, bladder cancer, liver cancer and biliary tract, kidney cancer, myeloid disorders, lymphoid disorders, Hodgkin, hairy cell, buccal cavity and pharynx (oral cavity), lip, tongue, mouth, pharynx, small intestine, colorectal, large intestine, rectum, brain and central nervous system, and leukemia, the method comprising administering IT-139 or a pharmaceutically acceptable composition thereof in combination with a chemotherapeutic agent.

[0120] According to another embodiment, the present invention relates to cancer selected from breast, ovary, cervix, prostate, testis, urogenital tract, esophagus, larynx, glioblastoma, neuroblastoma, stomach, skin, keratoacanthoma, lung, epidermoid carcinoma, large cell carcinoma, small cell carcinoma, lung adenocarcinoma, bone, colon, adenoma, pancreas, adenocarcinoma, thyroid, follicular carcinoma, undifferentiated carcinoma, papillary carcinoma, seminoma, melanoma, sarcoma, bladder cancer, liver cancer and biliary tract, kidney cancer, myeloid disorders, lymphoid disorders, Hodgkin, hairy cell, buccal cavity and pharynx (oral cavity), lip, tongue, mouth, pharynx, small intestine, colorectal, large intestine, rectum, brain and central nervous system, and leukemia A method for treating the same, the method comprising administering IT-139 or a pharmaceutically acceptable composition thereof in combination with a cancer immunotherapeutic agent.

[0121] Another embodiment provides a method for treating cancer by reducing the amount of GRP78 in cancer cells after administration of IT-139.

[0122] According to another embodiment, the present invention relates to a method for treating cancer by reducing the amount of GRP78 in cancer cells after administration of IT-139 in combination with a chemotherapeutic agent, wherein administration of IT-139 or a pharmaceutically acceptable composition thereof results in a greater reduction in the amount of GRP78 compared to administration of the chemotherapeutic agent.

[0123] According to another embodiment, the present invention provides a method for treating cancer by reducing the amount of GRP78 in cancer cells after administration of IT-139 in combination with a cancer immunotherapeutic agent, wherein administration of IT-139, or a pharmaceutically acceptable composition thereof, results in a greater reduction in the amount of GRP78 compared to administration of the cancer immunotherapeutic agent alone.

[0124] The order of administration of the therapeutic agents should be carefully considered. Without wishing to be bound by any particular theory, due to the mechanism of action and downregulation of GRP78, for maximum therapeutic effect, it is required to first administer any chemotherapeutic agent and then administer IT-139. As described above, treatment with a series of chemotherapeutic agents results in ER stress, which induces the production of GRP78. This process is a cell survival mechanism. Administration of IT-139 reduces the stress-induced levels of GRP78 that remove the cell survival pathway. The final result is an increase in cancer cell death and an increase in the antitumor effect.

[0125] According to one embodiment of the present invention, there is provided a method for treating cancer in a patient in need of cancer treatment, comprising: 1) administering a chemotherapeutic agent to the patient; 2) subsequently administering IT-139 or a pharmaceutically acceptable composition thereof to the patient; 3) optionally repeating steps 1 and 2.

[0126] In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered 1 day after the chemotherapeutic agent. In other embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered to the patient 1 day after the chemotherapeutic agent. In yet other embodiments, IT-139 is administered to the patient 1 to 7 days after the chemotherapeutic agent.

[0127] In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered concurrently with a chemotherapeutic agent. In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof and the chemotherapeutic agent are administered within about 20 to 28 hours of each other, or within about 22 to 26 hours of each other, or within about 24 hours of each other.

[0128] In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered prior to the chemotherapeutic agent. In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered at least about 8 to 16 hours before the chemotherapeutic agent, or at least about 10 to 14 hours before the chemotherapeutic agent, or at least about 12 hours before the chemotherapeutic agent.

[0129] In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered at least about 20 to 28 hours before the chemotherapeutic agent, or at least about 22 to 26 hours before the chemotherapeutic agent, or at least about 24 hours before the chemotherapeutic agent.

[0130] In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered at least about 44 to 52 hours before the chemotherapeutic agent, or at least about 46 to 50 hours before the chemotherapeutic agent, or at least about 48 hours before the chemotherapeutic agent.

[0131] In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered at least about 64 to 80 hours before the chemotherapeutic agent, or at least about 70 to 74 hours before the chemotherapeutic agent, or at least about 72 hours before the chemotherapeutic agent.

[0132] In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered before a chemotherapeutic agent. In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered at least about 8 to 16 hours after the chemotherapeutic agent, or at least about 10 to 14 hours after the chemotherapeutic agent, or at least about 12 hours after the chemotherapeutic agent.

[0133] In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered at least about 20 to 28 hours after the chemotherapeutic agent, or at least about 22 to 26 hours after the chemotherapeutic agent, or at least about 24 hours after the chemotherapeutic agent.

[0134] In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered at least about 44 to 52 hours after the chemotherapeutic agent, or at least about 46 to 50 hours after the chemotherapeutic agent, or at least about 48 hours after the chemotherapeutic agent.

[0135] In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered at least about 64 to 80 hours after the chemotherapeutic agent, or at least about 70 to 74 hours after the chemotherapeutic agent, or at least about 72 hours after the chemotherapeutic agent.

[0136] In certain embodiments, the chemotherapeutic agent is selected from the group consisting of gemcitabine, nanoparticle albumin paclitaxel, paclitaxel, docetaxel, cabazitaxel, oxaliplatin, cisplatin, carboplatin, doxorubicin, daunorubicin, sorafenib, everolimus, and vemurafenib. In certain embodiments, the chemotherapeutic agent is gemcitabine.

[0137] According to one embodiment of the present invention, there is provided a method for treating pancreatic cancer in a patient in need thereof, comprising: 1) administering gemcitabine and albumin nanoparticle paclitaxel; 2) Subsequently, administering IT-139 or a pharmaceutically acceptable composition thereof; and 3) Optionally, repeating steps 1 and 2 A method is provided that includes the above steps.

[0138] In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered concurrently with gemcitabine. In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof and gemcitabine are administered within about 20 to 28 hours of each other, or within about 22 to 26 hours of each other, or within about 24 hours of each other.

[0139] In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered prior to gemcitabine. In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered at least about 8 to 16 hours before gemcitabine, or at least about 10 to 14 hours before gemcitabine, or at least about 12 hours before gemcitabine.

[0140] In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered at least about 20 to 28 hours before gemcitabine, or at least about 22 to 26 hours before gemcitabine, or at least about 24 hours before gemcitabine.

[0141] In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered at least about 44 to 52 hours before gemcitabine, or at least about 46 to 50 hours before gemcitabine, or at least about 48 hours before gemcitabine.

[0142] According to one embodiment of the present invention, there is provided a method for treating cancer in a subject in need of cancer treatment, the method comprising administering IT-139 or a pharmaceutically acceptable composition thereof in combination with a cancer immunotherapeutic agent. In certain embodiments, the cancer immunotherapeutic agent is administered to the patient prior to the administration of IT-139 or a pharmaceutically acceptable composition thereof.

[0143] In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered simultaneously with the cancer immunotherapeutic agent. In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof, and the cancer immunotherapeutic agent are administered within about 20 to 28 hours of each other, or within about 22 to 26 hours of each other, or within about 24 hours of each other.

[0144] In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered prior to the cancer immunotherapeutic agent. In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered at least about 8 to 16 hours before the cancer immunotherapeutic agent, or at least about 10 to 14 hours before the cancer immunotherapeutic agent, or at least about 12 hours before the cancer immunotherapeutic agent.

[0145] In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered at least about 20 to 28 hours before the cancer immunotherapeutic agent, or at least about 22 to 26 hours before the cancer immunotherapeutic agent, or at least about 24 hours before the cancer immunotherapeutic agent.

[0146] In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered at least about 44 to 52 hours before the cancer immunotherapeutic agent, or at least about 46 to 50 hours before the cancer immunotherapeutic agent, or at least about 48 hours before the cancer immunotherapeutic agent.

[0147] In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered at least about 64 to 80 hours before, or at least about 70 to 74 hours before, or at least about 72 hours before a cancer immunotherapeutic agent.

[0148] In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered after a cancer immunotherapeutic agent. In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered at least about 8 to 16 hours after, or at least about 10 to 14 hours after, or at least about 12 hours after a cancer immunotherapeutic agent.

[0149] In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered at least about 20 to 28 hours after, or at least about 22 to 26 hours after, or at least about 24 hours after a cancer immunotherapeutic agent.

[0150] In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered at least about 44 to 52 hours after, or at least about 46 to 50 hours after, or at least about 48 hours after a cancer immunotherapeutic agent.

[0151] In certain embodiments, IT-139 or a pharmaceutically acceptable composition thereof is administered at least about 64 to 80 hours after, or at least about 70 to 74 hours after, or at least about 72 hours after a cancer immunotherapeutic agent.

[0152] In certain embodiments, the cancer immunotherapeutic agent is selected from the group consisting of cytokines, checkpoint inhibitors, and antibodies other than PD-1 antibodies. In certain embodiments, the cancer immunotherapeutic agent is selected from the group consisting of interferon, interleukin, PD-L1 antibody, alemtuzumab, ipilimumab, ofatumumab, atezolizumab, and rituximab.

[0153] According to one embodiment of the present invention, there is provided a method for treating cancer in a subject in need of cancer treatment, the method comprising administering IT-139 or a pharmaceutically acceptable composition thereof in combination with a PD-1 antibody. In certain embodiments, the PD-1 antibody is administered prior to the administration of IT-139 or a pharmaceutically acceptable formulation thereof.

[0154] According to one embodiment of the present invention, there is provided a method for treating cancer in a subject in need of cancer treatment, the method comprising administering IT-139 or a pharmaceutically acceptable composition thereof in combination with a PD-L1 antibody. In certain embodiments, the PD-L1 antibody is administered prior to the administration of IT-139 or a pharmaceutically acceptable formulation thereof.

[0155] According to one embodiment of the present invention, there is provided a method for treating cancer in a subject in need of cancer treatment, the method comprising administering IT-139 or a pharmaceutically acceptable composition thereof in combination with a cancer immunotherapeutic agent other than a PD-1 antibody. In certain embodiments, the cancer immunotherapeutic agent other than a PD-1 antibody is administered prior to the administration of IT-139 or a pharmaceutically acceptable formulation thereof.

Examples

[0156] The following examples are provided to enable a more thorough understanding of the invention described herein. These examples are for illustrative purposes only and should not be construed as limiting the invention in any way.

[0157] Analysis methods The following analysis methods were utilized to characterize the compounds of the present invention.

[0158] HPLC Method 1: The quantification and identity of sodium [trans - tetrachlorobis(1H - indazole)ruthenate(III)] were determined by high - pressure liquid chromatography with UV detection at 292 nm. The IT - 139 drug was dissolved in water at a concentration of 1 mg / mL, and 10 μL was injected into an Agilent Zorbax SB - C18 (3 μm, 4.6×150 mm) HPLC column. Mobile phase A consisted of 0.1% trifluoroacetic acid in water, and mobile phase B consisted of 0.1% trifluoroacetic acid in acetonitrile. Separation was achieved by a gradient flow of 1.0 mL / min such that mobile phase A changed from 90% to 10% from zero time to 12 minutes, and mobile phase B changed from 10% to 90% over 12 minutes. Then, from 12 minutes to 13 minutes, the gradient was reversed from 10% mobile phase A and 90% mobile phase B to 90% A and 10% B. This was continued until the end of the 15 - minute run. The analyte retention time was 7.7 minutes. The sample temperature was maintained at 5 °C and the column temperature was maintained at 25 °C.

[0159] HPLC Method 2: Since two different impurities co - elute when using HPLC Method 1, two HPLC methods are used. The related substances in the IT - 139 drug were determined by high - pressure liquid chromatography with UV detection at 292 nm. The IT - 139 drug was dissolved in water at a concentration of 1 mg / mL, and 10 μL was injected into an Agilent Zorbax SB - C18 (3 μm, 4.6×150 mm) HPLC column. Mobile phase A consisted of 0.1% trifluoroacetic acid in water, and mobile phase B consisted of 0.1% trifluoroacetic acid in acetonitrile. Separation was achieved by a gradient flow of 1.0 mL / min such that mobile phase A changed from 90% to 10% from zero time to 15 minutes, and mobile phase B changed from 10% to 90% over 15 minutes. The gradient was held at 10% A and 90% B from 15 minutes to 19.9 minutes. Then, from 19.9 minutes to 20 minutes, the gradient was reversed to 90% A and 10% B and held until the end of the 26 - minute run. The sample temperature was maintained at 5 °C and the column temperature was maintained at 25 °C.

[0160] HPLC Method 3: Analysis without decomposing Formula I-b from Compound A, Compound B, and Compound C uses high-performance liquid chromatography with UV detection at 297 nm. The IT-139 drug substance was dissolved in water at a concentration of 0.5 mg / mL, and 10 μL was injected into a Phenomenex Luna, Phenyl-Hexyl (150×3 mm×3 μm) HPLC column. The mobile phase consisted of 20 mM ammonium acetate buffer and 25% (volume / volume) methanol in 4 mM acetic acid. The separation was achieved by maintaining the column temperature at 25 °C and performing an isocratic flow of 1.0 mL / min for a total run time of 27 minutes.

[0161] Elemental Analysis: Galbraith Laboratories (Knoxville, TN) performed all elemental analysis measurements. Analysis of carbon, hydrogen, and nitrogen was carried out using standard procedure ME-14 (requiring weighing 1 - 5 mg and placing it in a tin capsule, then burning at 920 - 980 °C in a PerkinElmer 2400 Series II CHNS / O analyzer). Analysis of sodium and ruthenium was performed by inductively coupled plasma atomic emission spectrometry using standard procedures ME-70 and an ICP-OES Optima 5300 instrument. Analysis of cesium was carried out by inductively coupled plasma atomic emission spectrometry using standard procedure ME-30.

[0162] X-ray Diffraction: X-ray data was collected on a Bruker D8 Venture single-crystal diffractometer equipped with a PHOTON 100 CMOS detector, an IμS copper MX source, and an Oxford Cryostream Plus cryogenic device, or on a Bruker Smart Apex2 single-crystal diffractometer using copper radiation for room-temperature data collection.

[0163] Example 1 Purification of Ruthenium Chloride RuCl3.xH2O (100.0 g) was added to 600 mL of concentrated HCl and 99% ethanol It was combined with 600 mL. The mixture was distilled under normal pressure air to reduce the volume of the mixture to less than 400 mL. Then, the resulting concentrated ruthenium chloride solution remaining in the distillation flask was cooled to ambient temperature, filtered through a medium-porosity glass Buchner funnel, the Buchner funnel and the flask were rinsed with concentrated HCl, and the combined filtrate was diluted with additional concentrated HCl to a total volume of about 500 mL.

[0164] Example 2 Preparation of Indazolium Salt

Chemical formula

[0165] Example 3 Preparation of the Cesium Salt

Chemical formula

[0166] Example 4 Preparation of Sodium Salt

Chemical formula

[0167] Example 5 Removal of Residual Cesium 190.4 g of the material from Example 4 was transferred to a dry 10 L flask. Activated 4A molecular sieve powder (191 g) of equal weight was added. [Aldrich 688363-1KG, sodium aluminosilicate, "SYLOSIV A4" (manufactured by Grace Davidson)]. Methyl ethyl ketone (4.2 L) was added to the flask and the mixture was mechanically stirred. Methanol (600 mL) was added to the stirred slurry in 5 portions It was added slowly over time. Stirring (800 rpm) was continued for 30 minutes, during which time almost all of the dark brown lumps of material dissolved. The resulting orange slurry was filtered through a Whatman glass fiber GF - B filter disk placed on top of a fine - pore glass Buchner funnel. The spent molecular sieves were rinsed with an additional 0.4 L (2×200 mL) of MEK and discarded. The combined filtrate was precipitated by slowly adding 10 L of MTBE with mechanical stirring. Stirring was stopped and the mixture was left to settle for 30 minutes. The precipitated product was collected by filtration (3 L Buchner funnel), thoroughly rinsed with 1 L (2×0.5 L) of MTBE, and dried by suction for about 2 hours until the Buchner funnel became cold. This gave 184 g of the purified sodium salt. To remove traces of the solvent, the purified material was treated with wet MTBE. 184 g of the purified sodium salt was combined with 3.3 L of wet MTBE (water - saturated MTBE) in a 5 L wide - mouth Erlenmeyer flask and mechanically stirred for 40 minutes (200 rpm). The resulting brown solid was collected by filtration. The solid was rinsed with wet MTBE, dried by suction, and then thoroughly dried in vacuo overnight (15 hours). The product obtained was 176.11 g of a heavy, granular, coffee - brown solid with a purity of 98.7% as determined by HPLC (method 1). This corresponds to 85% of the total yield from RuCl3.xH2O (starting from Example 1). Elemental analysis determined that 35 - 750 ppm of cesium remained.

[0168] Example 6 Solution Stability Test The compound from Example 5 was prepared at room temperature (20 °C) using a room temperature solution and refrigerated (2 - 8 °C) using a cold (2 - 8 °C) solution. A total volume of 500 mL was used for each. A citric acid solution was prepared by dissolving 19.2 grams of citric acid in 1 L of water. A sodium citrate solution was prepared by dissolving 29.4 grams of sodium citrate in 1 L of water. The sodium citrate solution was added to the citric acid solution until the pH increased from 2.0 to 3.4. A mannitol solution was prepared by dissolving 13.3 g of mannitol in 200 mL of water. A solution of Formula I-b was prepared by adding 16.6 mL of citric acid buffer to 400 mL of water. 3.33 g of sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] was added to the solution and stirred for 10 minutes. 50 mL of the mannitol solution was added, followed by 33.4 mL of water to give a final volume of 500 mL of the IT-139 bulk solution. The room temperature (18 - 22 °C) samples were stirred using a magnetic stir plate on a laboratory workbench and the refrigerated samples were stirred using a magnetic stir plate in a refrigerator (2 - 8 °C). 100 μL aliquots were taken from each sample immediately after dissolution (T = 0) and at 0.5, 1, 2, 3, 4, 5, 6, 18, 24, 32, and 48 hours. Each sample was added to 1.9 mL of methanol in an HPLC vial and mixed by vortexing. The purity of sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] in the bulk solution stored at room temperature (18 - 22 °C) decreased from 96.8% to 12.1% after 18 hours. The purity of sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] in the bulk solution stored refrigerated (2 - 8 °C) decreased from 97.05% to 95.4% after 18 hours and to 89.8% after 48 hours. Figure 1 shows the percentage of sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] over 18 hours at room temperature (18 - 22 °C) and over 48 hours refrigerated (2 - 8 °C).Table 7 shows the percentages of sodium [tetrachlorobis(1H-indazole)ruthenate(III)] (RT 7.7 minutes) and impurities (total of RRT 0.7, 1.9, and unspecified RRT) in each sample based on HPLC peak area. The HPLC chromatograms of the IT-139 samples stored at refrigerated or room temperature at the 18-hour time point are shown in Figures 2 and 3, respectively.

Table 7

[0169] Example 7 Preparation of Compound A

Chemical formula

[0170] Example 8 Preparation of Compound C

Chemical formula

[0171] Example 9 Preparation of Compound D

Chemical formula

[0172] Example 10 IT-139 Formulation Process IT-139 was prepared in a cooled state using a cold (2 - 8 °C) solution to make a chemical with a total volume of 1 L. A citrate buffer was prepared by adding a sodium citrate solution (29.4 g / L in water) to a citric acid solution (19.2 g / L in water) until the pH increased from 2.0 to 3.4. Subsequently, 33 mL of the citrate buffer was added to 767 mL of water to prepare a working citrate buffer solution. 6.66 grams of sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] was added to 800 mL of the cold (2 - 8 °C) working citrate buffer solution, and the solution was stirred for 20 minutes while cooling to 2 - 8 °C using a magnetic stirring plate and a stir bar. A working solution of mannitol was prepared by dissolving 13.3 g of mannitol in 200 mL of water. 100 mL of the cold (2 - 8 °C) mannitol solution was added to the 800 mL of sodium trans-[tetrachlorobis(1H-indazole)ruthenate(III)] dissolved in the working citrate buffer, and the solution was stirred for 5 minutes while cooling to 2 - 8 °C. 100 mL of cold (2 - 8 °C) water was added to the chemical solution to make a final volume of 1 L. The IT-139 bulk chemical solution was filtered through a 0.22 μm syringe filter, aseptically filled into lyophilization vials, and made to a final fill volume of 30 mL in 50 mL clear glass vials. The vials were partially stoppered and loaded into a lyophilizer with the shelf pre-cooled to -5 °C. The lyophilization cycle consisted of freezing at -40 °C for 3 hours, primary drying at 0.1 mbar and -10 °C for 15 hours, then at 0.1 mbar and -5 °C for 10 hours, and secondary drying at 0.05 mbar and 5 °C for 2 hours, then at 0.05 mbar and 10 °C for 2 hours, then at 0.05 mbar and 15 °C for 2 hours, then at 0.05 mbar and 20 °C for 2 hours, for a total drying time of 36 hours. The vials were fully stoppered, sealed, and stored at -20 °C.

[0173] Example 11 IT-139 Formulation Process 2 IT-139 was prepared in a cooled state using a cold (2 - 8 °C) solution and made into a chemical with a total volume of 2.8 L. A citrate buffer was prepared by adding a sodium citrate solution (29.4 g / L in water) to a citric acid solution (19.2 g / L in water) until the pH increased from 2.0 to 3.4. Subsequently, 9.3 g of the citrate buffer was added to 1960 g of water to prepare a working citrate buffer solution. 36.3 g of sodium trans - [tetrachlorobis(1H - indazole)ruthenate(III)] was added to 1969.3 grams of the cold (2 - 8 °C) working citrate buffer solution, and the solution was stirred for 20 minutes while cooling to 2 - 8 °C using a magnetic stirring plate and a stir bar. A working solution of mannitol was prepared by dissolving 35 g of mannitol in 525 mL of water. 525 mL of the cold (2 - 8 °C) mannitol solution was added to the solution of sodium trans - [tetrachlorobis(1H - indazole)ruthenate(III)] in the working citrate buffer, and the solution was stirred for 5 minutes while cooling to 2 - 8 °C. 300 mL of cold (2 - 8 °C) water was added to the chemical solution to obtain a final volume of 2.8 L. The IT - 139 chemical bulk solution was filtered through a 0.22 μm syringe filter, aseptically filled into lyophilization vials, and had a final fill volume of 25.1 grams in 50 mL clear glass vials. The vials were partially stoppered and loaded into a lyophilizer with the shelf pre - cooled to - 5 °C. The lyophilization cycle consisted of freezing at - 40 °C for 6 hours, primary drying at 0.2 mbar and - 10 °C for 50 hours, and secondary drying at 0.2 mbar and 30 °C for 33 hours. The vials were backfilled with nitrogen, fully stoppered, sealed, and stored at 4 °C.

[0174] Example 12 Batch Analysis of IT - 139 API Sodium trans - [tetrachlorobis(1H - indazole)ruthenate(III)] and Ru III Cl3(Hind)2(H2O) and Ru III Cl3(Hind)2(CH3CN) and Ru IIITable 8 reproduces the batch analysis data in the drug substance containing Cl3(Hind)(HN=C(Me)ind) and cesium.

Table 8-1

Table 8-2

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Claims

1. A compound of formula Ia: 【Chemistry 21】 A method for preparing The method includes reacting trans-[tetrachlorobis(1H-indazole)ruthenate(III)]indazolium with cesium chloride in a solvent; isolating said compound of formula I-a by precipitation and filtration to obtain an isolated compound of formula I-a; and drying said isolated compound of formula I-a to obtain a dry powdered compound of formula I-a; wherein said compound of formula Ia is prepared by reacting trans-[tetrachlorobis(1H-indazole)ruthenate(III)]indazolium with cesium chloride in said solvent to obtain the synthesized compound of formula Ia; wherein the solvent is an alcohol having 1 to 5 carbon atoms, a diol having 2 to 4 carbon atoms, water, a ketone having 1 to 6 carbon atoms, a cyclic ether having 4 to 7 carbon atoms, an amide having 1 to 4 carbon atoms, DMSO, sulfolane, an ester having 4 to 6 carbon atoms, a chlorinated hydrocarbon having 1 or 2 carbon atoms, a liquid aromatic hydrocarbon, a nitrile having 2 to 6 carbon atoms, or a mixture thereof; method.

2. The method of claim 1, wherein the solvent comprises ethanol and methyl ethyl ketone (MEK) in an ethanol-methyl ethyl ketone (MEK) mixture, and the step of reacting trans-[tetrachlorobis(1H-indazole)ruthenate(III)]indazolium with cesium chloride forms a crystalline MEK solvate of the compound of formula I-a, and further comprises treating the crystalline MEK solvate of the compound of formula I-a with aqueous ethanol to obtain a hydrate form of the compound of formula I-a.

3. The method according to claim 1 or 2, wherein the step of reacting trans-[tetrachlorobis(1H-indazole)ruthenate(III)]indazolium with cesium chloride in a solvent includes reacting trans-[tetrachlorobis(1H-indazole)ruthenate(III)]indazolium with 1 to 10 equivalents of cesium chloride.

4. The method of claim 1, comprising reacting a compound of formula Ib: 【Chemical 22】 The method further comprises the step of preparing The method of claim 1, wherein the preparing step comprises mixing the compound of formula Ia with sodium aluminum sulfate (NaAl(SO 4 ) 2 ) in water and reacting the synthesized compound of formula Ia under conditions which result in a salt exchange, wherein the concentration of sodium aluminum sulfate in water is between 0.5M and 1.65M.

5. The method of claim 4, further comprising the steps of isolating the insoluble aluminum cesium sulfate and the compound of formula I-b by filtration to obtain an isolated compound of formula I-b, and further comprising the steps of dissolving the isolated compound of formula I-b in a dissolution solvent and removing the aluminum cesium sulfate by filtration to obtain the dissolved compound of formula I-b, wherein the dissolution solvent comprises an alcohol having 1 to 5 carbon atoms, a ketone having 3 to 6 carbon atoms, a nitrile having 2 to 5 carbon atoms, an ester having 3 to 6 carbon atoms, an amide having 1 to 4 carbon atoms, water, a diol having 1 to 4 carbon atoms, DMSO, sulfolane, or a combination thereof.

6. The method of claim 5, wherein the dissolution solvent comprises acetonitrile.

7. The method of claim 6, further comprising precipitating the dissolved compound of formula I-b with an anti-solvent to obtain a precipitated compound of formula I-b, wherein the anti-solvent comprises an ether having 3 to 8 carbon atoms, a cyclic, acyclic, or aromatic hydrocarbon having 5 to 8 carbon atoms, a chlorinated hydrocarbon having 1 to 4 carbon atoms, benzotrifluoride, chlorobenzene, methyl carbonate, or a mixture thereof.

8. The method of claim 7, wherein the anti-solvent comprises methyl tert-butyl ether (MTBE).

9. The method of claim 4, wherein the mixing is performed at 20°C to 25°C.

10. The method of claim 9, wherein the mixing is carried out for 12 hours to 168 hours.

11. The method of claim 4, further comprising removing residual cesium from the compound of formula I-b by stirring the compound of formula I-b with methanol in the presence of 4 Å molecular sieves, followed by precipitation with methyl tert-butyl ether (MTBE).

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