Processes for preparing AG-10, its intermediates, and salts thereof

Abstract

To propose improved processes of synthesizing AG-10, and provide other forms of AG-10 that offer advantageous pharmacokinetic properties.SOLUTION: Provided herein are: improved processes for the preparation of a compound of Formula IX; pharmaceutically acceptable salts of Formula I and Formula Ib; and crystalline forms of Formula IX (AG-10). The processes described herein provide improved yields and efficiency, while the pharmaceutically acceptable salts and crystalline forms thereof provide unexpected pharmacokinetic properties. Other features and aspects of the present disclosure will be apparent to a person skilled in the art upon reading the remainder of the specification.SELECTED DRAWING: Figure 1

Description

[Background technology]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application is a continuation of U.S. Provisional Application No. 62 / 460,576, filed February 17, 2017, under U.S. 35 U.S.C. § 119 ( Priority is claimed under paragraph e) of the present application, the disclosure of which is incorporated herein by reference in its entirety.

[0002] [Statement regarding rights to inventions obtained through federally sponsored research and development] Not applicable.

[0003] [Reference to a "Sequence Listing," table, or computer program listing on a compact disc, submitted as an appendix] Not applicable.

[0004] BACKGROUND OF THEINVENTION Aberrant protein interactions and aggregation through protein misfolding and excessive activation of signaling pathways are the underlying causes of many human degenerative diseases, therefore targeting protein-protein interactions (PPIs) is of therapeutic importance.

[0005] To date, the PPI inhibitors approved are proteins rather than small molecule inhibitors. For example, therapeutic monoclonal antibodies (mAbs) are used to treat cancer, autoimmune, infectious diseases, and neurodegenerative diseases. Therapeutic mAbs are expensive to produce, require administration by injection, and can elicit an immune response in patients. For these reasons, the development of small molecule inhibitors of PPIs is of interest.

[0006] One example of abnormal protein aggregation is the soluble protein transthyretin (TTR or prealbumin). TTR is a 55 kDa homotetrameric protein present in blood and cerebrospinal fluid. Upon dissociation from its homotetrameric form, TTR dimers can misfold into amyloidogenic monomers. This has been observed with wild-type TTR as well as over 100 different mutants. Previous studies have shown that stabilizing the tetrameric form of TTR suppresses the misfolding of the amyloidogenic monomer and subsequent TTR amyloid formation.

[0007] Recent studies have identified 3-(3-(3,5-dimethyl-1H-pyrazol-4-yl)propoxy)-4-fluorobenzoic acid (AG-10) as a promising candidate for treating TTR amyloid-related diseases, such as TTR amyloid cardiomyopathy. This compound is disclosed in WO 2014 / 100227. Notably, this disclosure does not provide any additional forms of AG-10, and the described synthetic method would not be suitable for industrial production.

[0008] Thus, there exists a need to propose improved methods for the synthesis of AG-10, and to provide other forms of AG-10 that impart advantageous pharmacokinetic properties. The present invention addresses these needs and provides related advantages as well. Summary of the Invention

[0009] 〔overview〕 In one aspect, the present disclosure provides an improved method for preparing a compound of formula IX, comprising:

[0010] [ka]

[0011] (a) a compound of formula II

[0012] [ka]

[0013] with a compound of formula III

[0014] [ka]

[0015] with a first base and a first organic solvent to provide a compound of formula IV

[0016] [ka]

[0017] (where each R 1 are independently a halogen or a sulfonate ester; (b) contacting the compound of formula IV with hydrazine and a second organic solvent to provide a compound of formula V.

[0018] [ka]

[0019] (c) contacting the compound of formula V with a sulfonating or halogenating agent to provide a compound of formula VI.

[0020] [ka]

[0021] (where R 2 is a halogen or a sulfonate ester);

[0022] (d) reacting a compound of formula VI with a compound of formula VII

[0023] [ka]

[0024] with a second base and a third organic solvent to provide a compound of formula VIII.

[0025] [ka]

[0026] (where R 3 is C 1 ~C 12 Alkyl, C 2 ~C 12 Alkenyl, C 1 ~C 12 Alkynyl, C 3 ~C 8 selected from the group consisting of cycloalkyl, heterocycloalkyl, aryl, and heteroaryl, each of which is optionally substituted; (e) contacting the compound of formula VIII with a third base to provide a compound of formula IX. The present invention provides a method comprising:

[0027] In a second aspect, the disclosure provides a pharma- ceutically acceptable salt represented by formula I or Ib:

[0028] [ka]

[0029] where X is a pharma- ceutically acceptable anion of a protic acid and Y is a multiprotic acid.

[0030] In a third aspect, the present disclosure discloses crystalline forms A-K of formula IX.

[0031] Other features, elements and aspects of the present disclosure will be apparent from the accompanying drawings and the detailed description that follows. [Brief description of the drawings]

[0032] [Figure 1] FIG. 1 shows the scheme described herein for the preparation of AG-10 and its intermediates. [Fig. 2A-2F] 2A-2F show pharmacokinetic results demonstrating the high bioavailability of AG-10 in multiple species. [Figure 3A-3F] 3A-3F show the pharmacokinetic results demonstrating the high bioavailability of AG-10 in male and female dogs at different doses. [Figure 4] FIG. 4 shows the X-ray powder diffraction (XRPD) pattern of the mesylate salt of formula IX. [Diagram 5] FIG. 5 shows the thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) plots of the mesylate salt of Formula IX. [Figure 6] FIG. 6 shows the X-ray powder diffraction (XRPD) pattern of the edisylate salt of formula IX. [Figure 7] FIG. 7 shows the thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) plots of the edisylate salt of formula IX. [Figure 8] FIG. 8 shows the X-ray powder diffraction (XRPD) pattern of the besylate salt of Formula IX. [Figure 9] FIG. 9 shows the thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) plots of the besylate salt of Formula IX. [Figure 10] FIG. 10 shows the X-ray powder diffraction (XRPD) pattern of the tosylate salt of Formula IX. [Figure 11] FIG. 11 shows the thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) plots of the tosylate salt of Formula IX. [Figure 12] FIG. 12 shows the X-ray powder diffraction (XRPD) pattern of the esylate salt of Formula IX. [Figure 13] FIG. 13 shows the thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) plots of the esylate salt of Formula IX. [Figure 14] FIG. 14 shows the X-ray powder diffraction (XRPD) pattern of the bromide salt of Formula IX. [Figure 15] FIG. 15 shows the thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) plots of the bromide salt of Formula IX. [Figure 16]FIG. 16 shows the X-ray powder diffraction (XRPD) pattern of Form a of the nitrate salt of Formula IX. [Figure 17] FIG. 17 shows thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) plots of Form a of the nitrate salt of Formula IX. [Figure 18] FIG. 18 shows the X-ray powder diffraction (XRPD) pattern of form b of the nitrate salt of Formula IX. [Figure 19] FIG. 19 shows the X-ray powder diffraction (XRPD) pattern of the sulfate salt of Formula IX. [Figure 20] FIG. 20 shows the thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) plots of the sulfate salt of Formula IX. [Figure 21] FIG. 21 shows the X-ray powder diffraction (XRPD) pattern of the oxalate salt of Formula IX. [Figure 22] FIG. 22 shows the thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) plots of the oxalate salt of Formula IX. [Diagram 23] FIG. 23 shows the X-ray powder diffraction (XRPD) pattern of Form a of the maleate salt of Formula IX. [Figure 24] FIG. 24 shows thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) plots of Form a of the maleate salt of Formula IX. [Diagram 25] FIG. 25 shows the X-ray powder diffraction (XRPD) pattern of Form b of the maleate salt of Formula IX. [Figure 26] FIG. 26 shows thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) plots of Form b of the maleate salt of Formula IX. [Figure 27] FIG. 27 shows the X-ray powder diffraction (XRPD) pattern of the acetate salt of Formula IX. [Figure 28] FIG. 28 shows the thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) plots of the acetate salt of Formula IX. [Figure 29] FIG. 29 shows the X-ray powder diffraction (XRPD) pattern of the L-malate salt of formula IX. [Diagram 30] FIG. 30 shows the thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) plots of the L-malate salt of formula IX. [Diagram 31] FIG. 31 shows the X-ray powder diffraction (XRPD) patterns of crystalline Form A of Formula IX (three different samples). [Diagram 32] FIG. 32 shows the thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) plots of crystalline Form A of Formula IX. [Diagram 33] FIG. 33 shows thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) plots of crystalline Form A of Formula IX. [Diagram 34] FIG. 34 shows thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) plots of crystalline Form A of Formula IX. [Diagram 35] FIG. 35 shows a polarized light microscopy (PLM) image of crystalline form A of formula IX. [Diagram 36] FIG. 36 shows the asymmetric unit structure of crystalline Form A of Formula IX. [Figure 37] FIG. 37 shows the dynamic vapor sorption (DVS) data for crystalline Form A of Formula IX. [Figure 38] FIG. 38 shows the X-ray powder diffraction (XRPD) patterns of crystalline Form A of Formula IX before (bottom) and after (top) DVS. [Figure 39] FIG. 39 shows a diagram summarizing the interconversion between the identified crystalline forms A, B, C, E, G, H, I, and J. [Diagram 40] FIG. 40 shows the X-ray powder diffraction (XRPD) pattern of crystalline form B of Formula IX. [Diagram 41] FIG. 41 shows the thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) plots of crystalline Form B of Formula IX. [Diagram 42] Figure 42 shows X-ray powder diffraction (XRPD) patterns of crystalline Form B before heating (top), heated to 100°C (second from the top), heated to 170°C (second from the bottom), and reference crystalline Form I (bottom). Upon heating, Form B transforms to Form I. [Diagram 43] FIG. 43 shows the X-ray powder diffraction (XRPD) pattern of crystalline Form C of Formula IX. [Diagram 44] FIG. 44 shows thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) plots of crystalline Form C of Formula IX. [Diagram 45] FIG. 45 shows the X-ray powder diffraction (XRPD) patterns of crystalline form D of formula IX (top plot) and crystalline form F of formula IX (bottom plot). [Figure 46] FIG. 46 shows the X-ray powder diffraction (XRPD) pattern of crystalline Form E of Formula IX. [Figure 47] FIG. 47 shows the thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) plots of crystalline Form E of Formula IX. [Figure 48] Figure 48 shows the X-ray powder diffraction (XRPD) patterns of crystalline Form E of Formula IX before heating (top), heated to 195°C (middle) and reference crystalline Form I (bottom). Upon heating, Form E transforms to Form I. [Figure 49] FIG. 49 shows the X-ray powder diffraction (XRPD) pattern of crystalline form Form G of Formula IX. [Figure 50] FIG. 50 shows the thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) plots of crystalline Form G of Formula IX. [Figure 51] FIG. 51 shows the X-ray powder diffraction (XRPD) pattern of crystalline Form H of Formula IX. [Figure 52] FIG. 52 shows thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) plots of crystalline Form H of Formula IX. [Figure 53] FIG. 53 shows the 1H-NMR spectrum of crystalline Form H of Formula IX. [Figure 54] Figure 54 shows the X-ray powder diffraction (XRPD) patterns of crystalline Form H of Formula IX before heating (top), heated to 120°C (middle) and reference crystalline Form I (bottom). Upon heating, Form H transforms to Form I. [Figure 55] FIG. 55 shows the X-ray powder diffraction (XRPD) pattern of crystalline Form I of Formula IX. [Figure 56] FIG. 56 shows the thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) plots of crystalline Form I of Formula IX. [Figure 57]Figure 57 shows powder X-ray diffraction (XRPD) patterns of crystalline form I of formula IX (reference, top), before N2 purging (second from top), after 1.5 hours of N2 purging (second from bottom), and reference crystalline form B (bottom). Upon N2 purging, form I transforms to form B. [Figure 58] FIG. 58 shows a thermogravimetric analysis (TGA) plot of crystalline Form I of Formula IX. [Figure 59] FIG. 59 shows the X-ray powder diffraction (XRPD) pattern of crystalline Form J of Formula IX. [Figure 60] FIG. 56 shows the thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) plots of crystalline Form J of Formula IX. [Figure 61] FIG. 61 shows the 1H NMR spectrum of crystalline form Form J of Formula IX. [Figure 62] Figure 62 shows X-ray powder diffraction (XRPD) patterns of crystalline Form J of Formula IX before heating (top), heated to 130°C (second from the top), reference crystalline Form A (second from the bottom), and reference crystalline Form I (bottom). Upon heating, Form J transforms into a mixture of Forms A and I. [Figure 63] FIG. 63 shows the X-ray powder diffraction (XRPD) pattern of crystalline form K of Formula IX. [Figure 64] FIG. 64 shows the thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC) plots of crystalline Form K of Formula IX. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0033] Detailed Description I. Overview

[0034] The present disclosure provides, in part, improved methods for preparing compound of formula IX (AG-10) and its intermediates. The newly described methods provide high yields and improved potency.

[0035] A complete synthetic scheme is provided in the Summary of the Invention section and in Scheme 1 (Figure 1), however, one of skill in the art will appreciate that certain steps of the process are novel and may be carried out regardless of the origin of the starting materials or intermediates.

[0036] Pharmaceutically acceptable salts of formula I and formula Ib are also provided. Pharmaceutically acceptable salts of formula I and formula Ib have surprising pharmacokinetic properties that improve the bioavailability of the compound of formula IX. Without being bound by a particular theory, the pharmaceutically acceptable salts of formula I and formula Ib provide a protonated pyrazole on the compound of formula IX that is paired with the anion of a protonic acid or a multi-protonic acid. Unlike the pharmaceutically acceptable salts of formula I and formula Ib, salts prepared from alkali hydroxides such as NaOH, or zwitterions of the compound of formula IX do not provide the beneficial features described herein. In certain embodiments, the compound of formula I is represented by the compound of formula Ia, which is the HCl salt of formula I. II. Definition

[0037] The term "compound of formula IX" refers to 3-(3-(3,5-dimethyl-1H-pyrazol-4-yl)propoxy)-4-fluorobenzoic acid, also known as AG-10, i.e.,

[0038] [ka]

[0039] It refers to a compound having the formula:

[0040] As used herein, the terms "a," "an," or "the" not only include embodiments having a single element, but also encompass embodiments having multiple elements. For example, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. For example, "a cell" includes multiple cells, and reference to "the agent" includes reference to one or more agents known to those of skill in the art.

[0041] The term "alkyl" refers to a straight or branched chain, saturated aliphatic group having the indicated number of carbon atoms. Alkyl can contain any number of carbon atoms, e.g., C 1-2 , C 1-3 , C 1-4 , C 1-5 , C 1-6 , C 1-7 , C 1-8 , C 1-9 , C 1-10 , C 2-3 , C 2-4 , C 2-5 , C 2-6 , C 3-4 , C 3-5 , C 3-6 , C 4-5 , C 4-6 and C 5-6 For example, C 1-6 Alkyl includes, but is not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, hexyl, and the like. Alkyl can also refer to alkyl groups having up to 20 carbon atoms, such as, but not limited to, heptyl, octyl, nonyl, decyl, and the like. Alkyl groups can be substituted or unsubstituted. Particular substituents include hydroxyl, halogen, alkoxy, and amino groups. One skilled in the art will know that various substituents can be added to alkyl groups without departing from the teachings herein.

[0042] The term "alkenyl" refers to a straight or branched chain hydrocarbon having at least two carbon atoms and at least one double bond. Alkenyl can be any number of carbons, e.g., C 2 , C 2-3 , C 2-4 , C 2-5 , C 2-6 , C 2-7 , C 2-8 , C 2-9 , C 2-10 , C 3 , C 3-4 , C 3-5 , C 3-6 , C4 , C 4-5 , C 4-6 , C 5 , C 5-6 and C 6 The alkenyl group can have any suitable number of double bonds, for example, but not limited to, 1, 2, 3, 4, 5 or more double bonds. Examples of alkenyl groups include, but are not limited to, vinyl (ethenyl), propenyl, isopropenyl, 1-butenyl, 2-butenyl, isobutenyl, butadienyl, 1-pentenyl, 2-pentenyl, isopentenyl, 1,3-pentadienyl, 1,4-pentadienyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 1,3-hexadienyl, 1,4-hexadienyl, 1,5-hexadienyl, 2,4-hexadienyl, or 1,3,5-hexatrienyl. The alkenyl group, like the alkyl groups, can be substituted or unsubstituted.

[0043] The term "alkynyl" refers to a straight or branched chain hydrocarbon having at least two carbon atoms and at least one triple bond. Alkynyl can be any number of carbons, e.g., C 2 , C 2-3 , C 2-4 , C 2-5 , C 2-6 , C 2-7 , C 2-8 , C 2-9 , C 2-10 , C 3 , C 3-4 , C 3-5 , C 3-6 , C 4 , C 4-5 , C 4-6 , C 5 , C 5-6 , and C 6Examples of alkynyl groups include, but are not limited to, acetylenyl, propynyl, 1-butynyl, 2-butynyl, isobutynyl, sec-butynyl, butadiynyl, 1-pentynyl, 2-pentynyl, isopentynyl, 1,3-pentadiynyl, 1,4-pentadiynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 1,3-hexadiynyl, 1,4-hexadiynyl, 1,5-hexadiynyl, 2,4-hexadiynyl, or 1,3,5-hexatriynyl. The alkynyl group may be substituted or unsubstituted, similar to the alkyl groups described above.

[0044] The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic, fused bicyclic or bridged polycyclic ring assembly containing 3 to 12 ring atoms or the number of atoms indicated. Cycloalkyl can include any number of carbons, e.g., C 3-6 , C 4-6 , C 5-6 , C 3-8 , C 4-8 , C 5-8 , C 6-8 , C 3-9 , C 3-10 , C 3-11 , and C 3-12 Saturated monocyclic cycloalkyl rings include, for example, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. Cycloalkyl groups can also be partially unsaturated, having one or more double or triple bonds in the ring. Cycloalkyl can include saturated monocyclic C 3-8 When cycloalkyl, exemplary groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Cycloalkyl groups can be substituted or unsubstituted. Those skilled in the art will recognize that a number of different substituents can be included in the cycloalkyl group without departing from the teachings herein.

[0045] The term "heterocycloalkyl" refers to a saturated ring system having 3 to 12 ring members and 1 to 4 heteroatoms such as N, O, and S. Additional heteroatoms are also useful, including, but not limited to, B, Al, Si, and P. The heteroatoms may be oxidized, including, but not limited to, -S(O)- and -S(O)-. 2 -. Heterocycloalkyl groups can contain any number of ring atoms, for example, 3 to 6, 4 to 6, 5 to 6, 3 to 8, 4 to 8, 5 to 8, 6 to 8, 3 to 9, 3 to 10, 3 to 11, or 3 to 12 ring members. Any suitable number of heteroatoms can be included in the heterocycloalkyl group, for example, 1, 2, 3, or 4, or 1 to 2, 1 to 3, 1 to 4, 2 to 3, 2 to 4, or 3 to 4 heteroatoms. Heterocycloalkyl groups can be substituted or unsubstituted, similar to the cycloalkyl groups described above.

[0046] The term "aryl" refers to an aromatic ring system having any suitable number of ring atoms and any suitable number of rings. An aryl group can contain any suitable number of ring atoms, for example, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 ring atoms and 6-10, 6-12, or 6-14 ring members. Aryl groups can be monocyclic or fused to form bicyclic or tricyclic groups, or linked by bonds to form biaryl groups. Exemplary aryl groups include phenyl, naphthyl, and biphenyl. Other aryl groups include benzyl, which has a methylene linking group. Certain aryl groups have 6-12 ring members, such as phenyl, naphthyl, or biphenyl. Other aryl groups can have 6-10 ring members, such as phenyl or naphthyl. Some other aryl groups have 6 ring members, such as phenyl. Aryl groups can be substituted or unsubstituted, similar to the cycloalkyl groups described above.

[0047] The term "heteroaryl" refers to a monocyclic or fused bicyclic or tricyclic aromatic ring system containing 5 to 16 ring atoms, where 1 to 5 of the ring atoms are heteroatoms such as N, O, or S. Additional heteroatoms are also useful, including but not limited to B, Al, Si, and P. The heteroatoms may be oxidized, including but not limited to -S(O)- and -S(O)-. 2 -. Heteroaryl groups can contain any number of ring atoms, for example, 3-6, 4-6, 5-6, 3-8, 4-8, 5-8, 6-8, 3-9, 3-10, 3-11, or 3-12 ring members. Any suitable number of heteroatoms can be contained in the heteroaryl group, for example, 1, 2, 3, 4, or 5, 1-2, 1-3, 1-4, 1-5, 2-3, 2-4, 2-5, 3-4, or 3-5 heteroatoms. Heteroaryl groups can have 5-8 ring members and 1-4 heteroatoms, or 5-8 ring members and 1-3 heteroatoms, or 5-6 ring members and 1-4 heteroatoms, or 5-6 ring members and 1-3 heteroatoms. Heteroaryl groups can include groups such as pyrrole, pyridine, imidazole, pyrazole, triazole, tetrazole, pyrazine, pyrimidine, pyridazine, triazine, thiophene, furan, thiazole, isothiazole, oxazole, and isoxazole. Heteroaryl groups can be substituted or unsubstituted, similar to the cycloalkyl groups described above.

[0048] The term "halogen" refers to fluorine, chlorine, bromine and iodine.

[0049] The term "hydrated" refers to a chemical reagent that includes water. "Hydrated" in reference to the chemical transformation of step (a) refers to a chemical reagent that has a sufficient amount of water to complete the indicated chemical transformation. In certain embodiments, the hydrated reagent includes at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15% or 20% water by weight.

[0050] III. Disclosed Embodiments A. Methods for Preparing Compounds of Formula IX In one aspect, the present disclosure provides an improved method for preparing a compound of formula IX, comprising:

[0051] [ka]

[0052] (a) a compound of formula II

[0053] [ka]

[0054] with a compound of formula III

[0055] [ka]

[0056] a first base, and a first organic solvent to provide a compound of formula IV

[0057] [ka]

[0058] (where each R 1 are independently a halogen or a sulfonate ester; (b) contacting the compound of formula IV with hydrazine and a second organic solvent to produce a compound of formula V

[0059] [ka]

[0060] Provide; (c) contacting the compound of formula V with a sulfonating or halogenating agent to provide a compound of formula VI.

[0061] [ka]

[0062] (where R 2 is a halogen or a sulfonate ester); (d) reacting a compound of formula VI with a compound of formula VII

[0063] [ka]

[0064] with a second base and a third organic solvent to provide a compound of formula VIII.

[0065] [ka]

[0066] (where R 3 is C 1-12 Alkyl, C 2-12 Alkenyl, C 1-12 Alkynyl, C 3-8 selected from the group consisting of cycloalkyl, heterocycloalkyl, aryl and heteroaryl, each of which is optionally substituted; and (e) contacting the compound of formula VIII with a third base to provide a compound of formula IX.

[0067] Step (a) comprises reacting a first base and an organic solvent with a compound of formula II

[0068] [ka]

[0069] and a compound of formula III

[0070] [ka]

[0071] and contacting the compound of formula IV

[0072] [ka]

[0073] This includes providing

[0074] Those skilled in the art will recognize that compounds of formula IV have both nucleophilic and electrophilic sites and are capable of intramolecular transformation depending on the reaction conditions. For example, under certain conditions, the alcohol group of formula IV can add to one of the carbonyl carbons to form a six-membered ring (formula IVa). When the above addition is coupled with a subsequent elimination reaction, compounds of formula IV have the structure of formula IVb. It will be apparent to those skilled in the art that compounds of formula IV can exist as enol tautomers of formula IVc.

[0075] [ka]

[0076] The compounds of formula IV, formula IVa, formula IVb, and formula IVc interconvert and depending on the reaction conditions, varying concentrations of these species are present, in some embodiments, only a single species is present.

[0077] There are many suitable bases that can be used for this conversion. For example, in some embodiments, the first base is an alkali metal carbonate, an alkali metal bicarbonate, or a combination thereof. Alkali metal carbonates include, but are not limited to, Li 2 CO 3 , Na 2 CO 3 and K. 2 CO 3 and alkali metal bicarbonates include, but are not limited to, LiHCO 3 , NaHCO 3 and K.H.C.O. 3 In some embodiments, the alkali metal carbonate is K 2 CO 3It is.

[0078] The organic solvent in step (a) is one that adequately dissolves both the compounds of formula II and formula III in solution and is miscible with the base used. Those skilled in the art will recognize that there are numerous organic solvents that meet these specifications. In some embodiments, the first organic solvent is a polar organic solvent. In some embodiments, the polar organic solvent is selected from the group consisting of acetone, ethyl acetate, dichloromethane, tetrahydrofuran, dimethylformamide, and acetonitrile. In some embodiments, the organic solvent is acetone.

[0079] The conversion in step (a) also includes at least one equivalent of water to generate the hydroxyl-containing compound of formula IV. Often, this equivalent of water is provided by a reagent or solvent in the reaction, such as the first base or an organic solvent, rather than by direct addition of water. The use of a hydrated base in the chemical conversion in step (a) provides a highly efficient conversion. Thus, in some embodiments, the first base in the conversion in step (a) is a hydrated base (aqueous base). In some embodiments, the first base is a hydrated alkali metal carbonate. In some embodiments, the first base is a hydrated K 2 CO 3 It is.

[0080] The compound of formula III has two R 1 groups, each of which is independently selected from the group consisting of chloride, bromide, tosylate, and mesylate. 1 is the bromide.

[0081] Each R 1 The group acts as a leaving group in the conversion of step (a); however, one of skill in the art will recognize that other leaving groups are useful in the present invention without departing from the teachings herein.

[0082] In some embodiments, the compound of formula IV obtained in step (a) is used directly in the conversion of step (b) without purification.

[0083] Moving to step (b), the described chemical transformations afford a compound of formula IV

[0084] [ka]

[0085] Hydrazine (N 2 H 4 ) and a second organic solvent to provide a compound of formula V.

[0086] [ka]

[0087] One of ordinary skill in the art will appreciate that a wide variety of solvents can be used as the second organic solvent in this transformation. In some embodiments, the second organic solvent is a polar protic organic solvent. In some embodiments, the polar protic organic solvent is a C 1-8 -OH In some embodiments, the polar protic organic solvent is ethanol.

[0088] In some embodiments, the compound of formula V obtained in step (b) is used directly in the conversion of step (c) without purification.

[0089] The chemical transformation of step (c) involves the replacement of the hydroxyl moiety in the compound of formula V with a halogen or conversion of said hydroxyl moiety to a sulfonic acid ester to provide a compound of formula (VI).

[0090] [ka]

[0091] Although sulfonating or halogenating agents are specifically contemplated as starting materials for this transformation, those skilled in the art will recognize that numerous different leaving groups besides halogens and sulfonates are contemplated as suitable for use with R 2Therefore, R 2 Any starting material capable of generating a suitable leaving group at the position falls within the scope of the present invention.

[0092] There are many solvents that are suitable for this transformation; however, one of skill in the art will recognize that the solvent selected for this chemical transformation will depend on the choice of sulfonating or halogenating agent, as a particular solvent may not be suitable for all starting materials. For example, polar organic solvents are particularly useful when contacting the compound of formula V with a halogenating agent. In one embodiment, the polar organic solvent is 1,2-dichloroethane.

[0093] Halogenating agents useful in the conversion of step (c) include, but are not limited to, PBr 3 , PCl 3 , PCl 5 , SOBr 2 , PBr 5 , and SOCl 2 Sulfonating agents for the conversion of step (c) include, but are not limited to, mesyl chloride (MsCl) and tosyl chloride (TsCl). In some embodiments, the halogenating agent is PBr 3 It is.

[0094] R 2 The identity of depends on the starting material selected for the chemical transformation of step (c). For example, if a sulfonating agent is selected, R 2 is the corresponding sulfate. 2 is chloride, bromide, tosylate, and mesylate. 2 is Br. Turning attention to step (d), a compound of formula VI

[0095] [ka]

[0096] and a compound of formula VII

[0097] [ka]

[0098] in the presence of a base and a third organic solvent to provide a compound of formula VIII:

[0099] [ka]

[0100] The chemical conversion of step (d) can be carried out using a wide variety of bases. For example, in some embodiments, the second base is an alkali metal carbonate, an alkali metal bicarbonate, or a combination thereof. Alkali metal carbonates include, but are not limited to, Li 2 CO 3 , Na 2 CO 3 , and K 2 CO 3 and alkali metal bicarbonates include, but are not limited to, LiHCO 3 , NaHCO 3 , and KHCO 3 In some embodiments, the alkali metal carbonate is K 2 CO 3 It is.

[0101] The organic solvent in step (d) is one that adequately dissolves both the compounds of formula VI and formula VII in solution and is miscible with the base used. Those skilled in the art will recognize that there are numerous organic solvents that meet these specifications. In some embodiments, the third organic solvent is a polar aprotic organic solvent. In some embodiments, the polar organic solvent is selected from the group consisting of acetone, ethyl acetate, dichloromethane, tetrahydrofuran, dimethylformamide, dimethylsulfoxide, and acetonitrile. In some embodiments, the third organic solvent is dimethylformamide. In some embodiments, the third organic solvent is dimethylsulfoxide.

[0102] R 3 Suitable substituents for the group include those that do not interfere with the chemical transformations of step (e) described in detail below. Such substituents include, but are not limited to, C 1-8 Alkyl, C 3-8 Cycloalkyl, C 3-12 heterocycloalkyl, aryl, heteroaryl, etc. Those skilled in the art will appreciate that R 3 It will be appreciated that numerous other ester substituents of R are suitable without departing from the teachings herein. 3 C 1-8 In some embodiments, R 3 is methyl.

[0103] In some embodiments, the process of step (d) provides the compound of formula VIII in at least a 70% yield (mol / mol) with respect to the amount of formula VII.

[0104] With regard to step (e), the compound of formula VIII

[0105] [ka]

[0106] is contacted with a third base to provide a compound of formula IX

[0107] [ka]

[0108] The third base used in the chemical transformation of step (e) can be a number of different bases. For example, in some embodiments, the third base is a metal hydroxide. In some embodiments, the metal hydroxide is an alkali metal hydroxide. In some embodiments, the alkali metal hydroxide is selected from the group consisting of LiOH, NaOH, KOH, RbOH, and CsOH. In some embodiments, the alkali metal hydroxide is LiOH. In some embodiments, the alkali metal hydroxide is NaOH.

[0109] One of ordinary skill in the art will know that a wide variety of solvents can be used as the solvent for the conversion in step (e). For example, in some embodiments, the second organic solvent is a polar protic organic solvent or water. In some embodiments, the polar organic solvent is a C 1-8 In some embodiments, the polar protic organic solvent is methanol. In some embodiments, the solvent is water. In some embodiments, the solvent is a combination of methanol and water.

[0110] In some embodiments, the process of step (e) further comprises: (ei) removing the solvent to give a residue; (e-ii) dissolving the residue in water to form a solution; (e-iii) acidifying the solution to form a precipitate; and (e-iv) filtering the solution to provide isolated Formula IX wherein steps (ei) through (e-iv) are performed after step (e).

[0111] Step (ei) can be carried out using any suitable removal step, such as reduced pressure, elevated temperature, or a combination of both. In some implementations, the solvent is removed under reduced pressure. In some embodiments, a solid is produced in step (e) and the solvent is removed by filtration. Additionally, the addition of water in step (e-ii) can be carried out prior to step (ei). In such cases, the removal of the solvent under reduced pressure provides a concentrated aqueous component (i.e., water is not removed). Re-ordering of steps (ei) and (e-ii) does not depart from the scope of the methods described herein.

[0112] Step (e-iii) can be acidified using any suitable acid. In some embodiments, the suitable acid is HCl. In some embodiments, the solution is acidified to a pH of 3 or less, 0-3, or 2-3 or less. In some embodiments, the solution is acidified to a pH of about 2. In some embodiments, the solution is acidified to a pH of 2 or less, 0-2, or 1-2. In some embodiments, the solution is acidified to a pH of about 1.4-1.6.

[0113] The pH of the acidification step (e-iii) determines the predominant species produced. In some embodiments, the pH of the acidification step is in the range of 5-6 to produce the zwitterionic form of formula IX. In some embodiments, acidification with HCl to a pH of less than about 2 or in the range of 1.4-1.6 produces the HCl salt of formula IX (i.e., the compound of formula Ia).

[0114] The process described in step (e) can produce a compound of formula IX in high yield and purity. In some embodiments, the yield of step (e) is greater than 85%, 90%, 93% or 95% (mol / mol) based on the compound of formula VIII. In some embodiments, the purity of the compound of formula IX produced in step (e) is greater than 80%, 85%, 90%, 95% or 97% (mol / mol).

[0115] In another aspect, a method for preparing a compound of formula IX comprises the steps of:

[0116] [ka]

[0117] (a) a compound of formula II

[0118] [ka]

[0119] with a compound of formula III

[0120] [ka]

[0121] contacting the first base with a first organic solvent to provide an adduct; Here, each R 1 are independently a halogen or a sulfonate ester; (b) contacting the adduct with hydrazine and a second organic solvent to produce a compound of formula V Provide

[0122] [ka]

[0123] (c) contacting the compound of formula V with a sulfonating or halogenating agent to provide a compound of formula VI.

[0124] [ka]

[0125] (where R 2 is a halogen or a sulfonate ester); (d) reacting a compound of formula VI with a compound of formula VII

[0126] [ka]

[0127] with a second base and a third organic solvent to provide a compound of formula VIII.

[0128] [ka]

[0129] Here, R 3 is C 1-12 Alkyl, C 2-12 Alkenyl, C 1-12 Alkynyl, C 3-8 selected from the group consisting of cycloalkyl, heterocycloalkyl, aryl, and heteroaryl, each of which is optionally substituted; and (e) contacting the compound of formula VIII with a third base to provide a compound of formula IX. A method is provided that includes:

[0130] In some embodiments, the adduct produced in step (a) is a compound of Formula IV, Formula IVa, Formula IVb, and / or Formula IVc.

[0131] [ka]

[0132] One skilled in the art will appreciate that the compounds set forth above are interchangeable and that the relative amounts of each compound will depend on the experimental conditions.

[0133] As noted above, one of skill in the art will appreciate that certain steps of the method may be performed regardless of the origin of the starting materials or intermediates. B. Pharmaceutically Acceptable Salts of Formula I

[0134] In a second aspect, the present disclosure provides a pharma- ceutically acceptable salt represented by formula I:

[0135] [ka]

[0136] where X is a pharma- ceutically acceptable salt of a protic acid.

[0137] A variety of protic acids are suitable for preparing pharma- ceutically acceptable salts of Formula I. It is understood that the pharma- ceutically acceptable salts of a protic acid will depend on the protic acid used. For example, protic acids useful in the present disclosure include hydrochloric acid, hydrobromic acid, sulfonic acid, tosylic acid (p-toluenesulfonic acid), methanesulfonic acid, nitric acid, or acetic acid. Thus, pharma- ceutically acceptable anions of a protic acid include chloride (Cl), bromide (Br). - ), sulfonate ion (HS(O) 2 O - ), tosylate ion (TsO - ), mesylate ion (MsO - ), besylate ion (BeO - ), ethanesulfonate ion (EtSO 3 - ), nitrate ion (NO 3 - ), acetate ion (CH 3 C(O)O - ), glycolate ion (HO-CH 2 -C(O)O - ), or a combination thereof.

[0138] In some embodiments, the pharma- ceutically acceptable anion of the protic acid is mesylate. In some embodiments, the mesylate salt of Formula IX is characterized by an X-ray powder diffraction pattern substantially in accordance with FIG.

[0139] In some embodiments, the pharma- ceutically acceptable anion of the protic acid is besylate. In some embodiments, the besylate salt of Formula IX is characterized by an X-ray powder diffraction pattern substantially in accordance with FIG.

[0140] In some embodiments, the pharma- ceutically acceptable anion of the protic acid is tosylate. In some embodiments, the tosylate salt of Formula IX is characterized by an X-ray powder diffraction pattern substantially in accordance with FIG.

[0141] In some embodiments, the pharma- ceutically acceptable anion of the protic acid is esylate. In some embodiments, the esylate salt of Formula IX is characterized by an X-ray powder diffraction pattern substantially in accordance with FIG.

[0142] In some embodiments, the pharma- ceutically acceptable anion of the protic acid is bromide. In some embodiments, the bromide salt of formula IX is characterized by an X-ray powder diffraction pattern substantially in accordance with FIG.

[0143] In some embodiments, the pharma- ceutically acceptable anion of the protic acid is a nitrate ion. In some embodiments, the nitrate salt of Formula IX is characterized by an X-ray powder diffraction pattern substantially in accordance with FIG.

[0144] In some embodiments, the pharma- ceutically acceptable anion of the protic acid is chloride, and the pharma- ceutically acceptable salt of Formula I is represented by Formula (Ia):

[0145] [ka]

[0146] In some embodiments, the salt of Formula Ia is crystalline Form A. In some embodiments, crystalline Form A of Formula I is characterized by an X-ray powder diffraction pattern having peaks at 12.0, 21.8, 25.9, 26.7, and 27.9 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the X-ray powder diffraction pattern further comprises one or more peaks at 7.0, 10.3, 13.9, 15.6, and / or 17. Crystalline Form A of Formula Ia is characterized by an X-ray powder diffraction pattern substantially in accordance with FIG.

[0147] In some embodiments, a multiprotic acid, such as a diprotic acid or a triprotic acid, is used to prepare a pharma- ceutically acceptable salt of formula IX. In such embodiments, the pharma- ceutically acceptable salt is represented by formula Ib:

[0148] [ka]

[0149] (where Y is a multiprotic acid).

[0150] In some embodiments, Y is selected from the group consisting of ethane-1,2-disulfonic acid, sulfuric acid, citric acid, maleic acid, malic acid, tartaric acid, and oxalic acid, hi some embodiments, Y is L-malic acid or L-tartaric acid.

[0151] In some embodiments, Y is ethane-1,2-disulfonic acid. In some embodiments, the edisylate salt of Formula IX is characterized by an XRPD pattern substantially in accordance with FIG.

[0152] In some embodiments, Y is sulfate. In some embodiments, the sulfate salt of Formula IX is characterized by an XPRD pattern substantially in accordance with FIG.

[0153] In some embodiments, Y is oxalic acid. In some embodiments, the oxalate salt of Formula IX is characterized by an XPRD pattern substantially in accordance with FIG.

[0154] In some embodiments, Y is maleic acid. In some embodiments, the maleate salt of formula IX is characterized by an XPRD pattern substantially in accordance with Figure 23. In some embodiments, the maleate salt of formula IX is characterized by an XRPD pattern substantially in accordance with Figure 25.

[0155] In some embodiments, Y is acetate. In some embodiments, the acetate salt of Formula IX is characterized by an XRPD pattern substantially in accordance with FIG.

[0156] In some embodiments, Y is L-malic acid. In some embodiments, the L-malate salt of Formula IX is characterized by an XRPD pattern substantially in accordance with FIG.

[0157] The molar ratio of AG-10 to Y in formula Ib can vary depending on the multiprotic acid used, for example, when Y is maleic acid, the molar ratio of AG-10 to Y is 1:1, when Y is edisylate, the molar ratio of AG-10 to Y is 2:1, and when Y is malic acid, the molar ratio of AG-10 to Y is 1.8:1.

[0158] Pharmaceutically acceptable salts of formula I can be prepared using a number of conventional methods known in the art. For example, the free acid form of the compound of formula I can be contacted with a stoichiometric amount of a suitable acid in water, an organic solvent, or a mixture of the two. In some embodiments, the pharma- ceutically acceptable salts of formula I are formed in a non-aqueous medium such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile. In some embodiments, the pharma- ceutically acceptable salts of formula I are formed by dissolving the compound of formula IX in water, adding an appropriate amount of HX to the mixture, and then adding a non-aqueous solvent, such as the non-aqueous medium described above, to crystallize the salt. In some embodiments, the appropriate amount of HX is a stoichiometric amount. It is understood that HX comprises hydrogen and X is a pharma- ceutically acceptable anion of a protic acid as defined above.

[0159] As with the pharma- ceutically acceptable salts of formula I, pharma- ceutically acceptable salts of formula Ib can be prepared using a number of conventional methods known in the art. As a non-limiting example, the free acid form of the compound of formula Ib can be contacted with a stoichiometric or substoichiometric amount of an appropriate multiprotic acid in water, an organic solvent, or a mixture of the two, to prepare a pharma- ceutically acceptable salt of formula Ib. C. Crystalline Form of Formula IX

[0160] In another embodiment, a crystalline form of formula IX is provided

[0161] [ka]

[0162] The present disclosure describes eleven crystalline forms of Formula IX: six HCl salt forms (Forms A, B, E, H, I and J), three free base forms (Forms K, C and G), and two uncharacterized forms (Forms D and F). A summary of the properties of the identified forms is provided in Tables 1 and 2.

[0163] In some embodiments, the crystalline form of Formula IX provided herein is substantially free of other crystalline forms, the term "substantially free" referring to an amount of another crystalline form of 10% or less, preferably an amount of another crystalline form of 8%, 5%, 4%, 3%, 2%, 1%, 0.5% or less.

[0164] [Table 1]

[0165] [Table 2]

[0166] In some embodiments, crystalline Form A of formula IX is provided. In some embodiments, crystalline Form A of formula IX is characterized by an X-ray powder diffraction pattern including peaks at 7.0, 10.4, 12.0, 13.0, and 13.9 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Form A of formula IX is characterized by an X-ray powder diffraction pattern including peaks at 12.0, 21.8, 25.9, 26.7, and 27.9 degrees 2θ (±0.2 degrees 2θ). In some embodiments, the X-ray powder diffraction pattern further includes one or more peaks at 7.0, 10.3, 13.9, 15.6, and / or 17. In some embodiments, crystalline Form A of formula IX is characterized by an X-ray powder diffraction pattern substantially in accordance with Figure 31. In some embodiments, crystalline Form A of formula IX is substantially free of other crystalline forms.

[0167] In some embodiments, crystalline Form A of Formula IX is characterized by a weight loss, as measured by thermogravimetric analysis, in the range of about 0.7% to about 1.9% upon heating to about 150° C. In some embodiments, the weight loss, as measured by thermogravimetric analysis, is about 1.3%.

[0168] In some embodiments, crystalline Form A of formula IX is characterized by about 1.6% water uptake at 25° C. / 80% RH after undergoing dynamic vapor sorption cycling, including pre-equilibration at 0% relative humidity (RH). In some embodiments, crystalline Form A of formula IX is characterized by less than 2.5% weight gain after undergoing dynamic vapor sorption cycling from about 0% relative humidity (RH) to about 90% RH. In some embodiments, crystalline Form A of formula IX has a dynamic vapor sorption profile substantially as shown in FIG.

[0169] In some embodiments, crystalline Form A of Formula IX is characterized by a differential scanning calorimetry thermogram that includes endothermic peaks near 211-214° C. and 237-239° C. In some embodiments, the differential scanning calorimetry thermogram includes endothermic peaks near 11.7, 212.6, and 237.3° C.

[0170] In some embodiments, crystalline form B of formula IX is provided. In some embodiments, crystalline form B of formula IX is characterized by an X-ray powder diffraction pattern including peaks at 12.0, 13.8, 17.2, 17.7, and 19.8 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline form B of formula IX is characterized by an X-ray powder diffraction pattern including peaks at 12.1, 13.9, 19.8, 23.3, and 24.4 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline form B of formula IX is characterized by an X-ray powder diffraction pattern substantially in accordance with Figure 40. In some embodiments, crystalline form B of formula IX is substantially free of other crystalline forms.

[0171] In some embodiments, crystalline form B of formula IX is characterized by a weight loss in the range of about 0.6% to about 2.0% when heated to about 150° C., as measured by thermogravimetric analysis. Crystalline form B of formula IX is characterized by a weight loss of about 1.2% when heated to about 150° C., as measured by thermogravimetric analysis.

[0172] In some embodiments, crystalline form B of Formula IX is characterized by a differential scanning calorimetry thermogram comprising endothermic peaks at approximately 161.4°C, 232.2°C, and 262.3°C.

[0173] In some embodiments, crystalline Form E of formula IX is provided. In some embodiments, crystalline Form E of formula IX is characterized by an X-ray powder diffraction pattern including peaks at 11.8, 14.0, 15.1, 19.9, and 24.0 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Form E of formula IX is characterized by an X-ray powder diffraction pattern including peaks at 11.9, 14.0, 15.1, and 25.8 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Form E of formula IX is characterized by an X-ray powder diffraction pattern substantially in accordance with Figure 46. In some embodiments, crystalline Form E of formula IX is substantially free of other crystalline forms.

[0174] Crystalline Form E of Formula IX is characterized by a weight loss in the range of about 0.5% to about 2.5% when heated to about 150° C., as measured by thermogravimetric analysis. In some embodiments, crystalline Form E of Formula IX is characterized by a weight loss of about 1.5% when heated to about 150° C., as measured by thermogravimetric analysis.

[0175] In some embodiments, crystalline Form E of Formula IX is characterized by a differential scanning calorimetry thermogram comprising endothermic peaks near 182.0°C and 242.7°C.

[0176] In some embodiments, crystalline Form I of Formula IX is provided. In some embodiments, crystalline Form I of Formula IX is characterized by an X-ray powder diffraction pattern including peaks at 11.4, 12.1, 12.4, 13.6, and 13.9 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Form I of Formula IX is characterized by an X-ray powder diffraction pattern including peaks at 12.5, 17.3, 23.4, 25.0, and 25.4 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Form I of Formula IX is characterized by an X-ray powder diffraction pattern substantially in accordance with Figure 55. In some embodiments, crystalline Form I of Formula IX is substantially free of other crystalline forms.

[0177] In some embodiments, crystalline Form I of Formula IX is characterized by a weight loss in the range of about 2.5% to about 3.5% when heated to about 120° C., as measured by thermogravimetric analysis. In some embodiments, crystalline Form I of Formula IX is characterized by a weight loss of about 3.0% when heated to about 120° C., as measured by thermogravimetric analysis.

[0178] In some embodiments, crystalline Form I of formula IX is characterized by a differential scanning calorimetry thermogram comprising endothermic peaks at approximately 62.0°C, 158.4°C, and 215.7°C.

[0179] In some embodiments, crystalline Form H of formula IX is provided. In some embodiments, crystalline Form H of formula IX is characterized by an X-ray powder diffraction pattern including peaks at 11.8, 12.3, 13.8, 15.7, and 16.9 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Form H of formula IX is characterized by an X-ray powder diffraction pattern including peaks at 11.9, 12.3, 21.7, 23.3, and 25.8 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Form H of formula IX is characterized by an X-ray powder diffraction pattern substantially in accordance with Figure 51. In some embodiments, crystalline Form H of formula IX is substantially free of other crystalline forms.

[0180] In some embodiments, crystalline Form H of Formula IX is characterized by a weight loss in the range of about 3.5% to about 5.5% when heated to about 150° C., as measured by thermogravimetric analysis. In some embodiments, crystalline Form H of Formula IX is characterized by a weight loss of about 4.6% when heated to about 150° C., as measured by thermogravimetric analysis.

[0181] In some embodiments, crystalline Form H of Formula IX is characterized by a differential scanning calorimetry thermogram containing endothermic peaks near 90.4°C, 200.5°C, and 232.3°C.

[0182] In some embodiments, crystalline Form J of formula IX is provided. In some embodiments, crystalline Form J of formula IX is characterized by an X-ray powder diffraction pattern including peaks at 4.6, 11.8, 12.8, 13.8, and 14.6 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Form J of formula IX is characterized by an X-ray powder diffraction pattern including peaks at 13.8, 14.7, 22.9, 26.2, and 27.7 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Form J of formula IX is characterized by an X-ray powder diffraction pattern substantially in accordance with Figure 59. In some embodiments, crystalline Form J of formula IX is substantially free of other crystalline forms.

[0183] Crystalline form J of formula IX is characterized by a weight loss in the range of about 17.5% to about 24% when heated to about 120° C., as measured by thermogravimetric analysis. In some embodiments, crystalline form J of formula IX is characterized by a weight loss of about 21.5% when heated to about 120° C., as measured by thermogravimetric analysis.

[0184] In some embodiments, crystalline Form J of Formula IX is characterized by a differential scanning calorimetry thermogram comprising endothermic peaks at approximately 120.8°C, 197.8°C, and 221.5°C.

[0185] In some embodiments, crystalline form K of formula IX is provided. In some embodiments, crystalline form K of formula IX is characterized by an X-ray powder diffraction pattern including peaks at 7.5, 9.8, 13.9, 15.9, and 19.3 degrees 2θ (± 0.2 degrees 2θ). In some embodiments, crystalline form K of formula IX is characterized by an X-ray powder diffraction pattern including peaks at 7.2, 7.6, 9.9, 14.0, and 19.3 degrees 2θ (± 0.2 degrees 2θ). In some embodiments, crystalline form K of formula IX is characterized by an X-ray powder diffraction pattern substantially in accordance with Figure 59. In some embodiments, crystalline form K of formula IX is substantially free of other crystalline forms.

[0186] In some embodiments, crystalline form K of Formula IX is characterized by a weight loss in the range of about 5.0% to about 7.0% when heated to about 120° C., as measured by thermogravimetric analysis. In some embodiments, crystalline form K of Formula IX is characterized by a weight loss of about 6.1% when heated to about 120° C., as measured by thermogravimetric analysis.

[0187] In some embodiments, crystalline form K of Formula IX is characterized by a differential scanning calorimetry thermogram comprising endothermic peaks at approximately 159.3°C, 176.2°C, and 278.4°C.

[0188] In some embodiments, crystalline Form C of Formula IX is provided. In some embodiments, crystalline Form C of Formula IX is characterized by an X-ray powder diffraction pattern including peaks at 9.5, 11.7, 12.3, 13.4, and 14.6 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Form C of Formula IX is characterized by an X-ray powder diffraction pattern including peaks at 14.6, 16.8, 19.5, 20.7, and 22.5 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline Form C of Formula IX is characterized by an X-ray powder diffraction pattern substantially in accordance with Figure 43. In some embodiments, crystalline Form C of Formula IX is substantially free of other crystalline forms.

[0189] In some embodiments, crystalline Form C of Formula IX is characterized by a weight loss in the range of about 2.0% to about 4.0% when heated to about 150° C., as measured by thermogravimetric analysis. In some embodiments, crystalline Form C of Formula IX is characterized by a weight loss of about 3.1% when heated to about 150° C., as measured by thermogravimetric analysis.

[0190] In some embodiments, crystalline form C of formula IX is characterized by a differential scanning calorimetry thermogram comprising endothermic peaks near 91.2°C and 173.0°C.

[0191] In some embodiments, crystalline form G of formula IX is provided. In some embodiments, crystalline form G of formula IX is characterized by an X-ray powder diffraction pattern including peaks at 9.8, 12.2, 13.1, 13.4, and 14.6 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline form G of formula IX is characterized by an X-ray powder diffraction pattern including peaks at 12.3, 13.2, 13.4, 17.8, and 26.6 degrees 2θ (±0.2 degrees 2θ). In some embodiments, crystalline form G of formula IX is characterized by an X-ray powder diffraction pattern substantially in accordance with Figure 43. In some embodiments, crystalline form G of formula IX is substantially free of other crystalline forms.

[0192] In some embodiments, crystalline form G of formula IX is characterized by a weight loss in the range of about 1.7% to about 2.7% when heated to about 200° C., as measured by thermogravimetric analysis. In some embodiments, crystalline form G of formula IX is characterized by a weight loss of about 3.7% when heated to about 200° C., as measured by thermogravimetric analysis.

[0193] In some embodiments, crystalline form Form G of Formula IX is characterized by a differential scanning calorimetry thermogram comprising an endothermic peak at about 231.1°C.

[0194] In some embodiments, crystalline Form D of Formula IX is provided. In some embodiments, crystalline Form D of Formula IX is characterized by a powder X-ray diffraction pattern substantially in accordance with Figure 45 (top plot). In some embodiments, crystalline Form D of Formula IX is substantially free of other crystalline forms.

[0195] In some embodiments, crystalline Form F of formula IX is provided. In some embodiments, crystalline Form F of formula IX is characterized by a powder X-ray diffraction pattern substantially in accordance with Figure 45 (lower plot). In some embodiments, crystalline Form F of formula IX is substantially free of other crystalline forms.

[0196] Methods for producing the described crystalline forms are described in further detail in the Examples herein. Crystallization conditions used to produce Forms A-K include anti-solvent addition, slow evaporation, slow cooling, slurry conversion at room temperature, slurry conversion at 50°C, solid-phase vapor diffusion, and liquid-phase vapor diffusion. EXAMPLES

[0197] IV. Working Examples Example 1: Preparation of 3-(3-hydroxypropyl)pentane-2,4-dione (compound of formula IV)

[0198] [ka]

[0199] The compound of formula IIIa (100 g, 495 mmol, 1.0 equiv.) was dissolved in acetone (1 L). To the solution was added the compound of formula II (49.59 g, 495 mmol, 1.0 equiv.), followed by addition of K 2 CO 3 (82.14 g, 594.38 mmol, 1.2 equiv.) and KI (41.11 g, 247 mmol, 0.5 equiv.) were added with stirring at room temperature. The reaction mixture was heated to 60±5° C. and stirred at this temperature for 40 h. The reaction mixture was filtered and then concentrated under reduced pressure to give the compound of formula IV (102 g) as a viscous orange liquid.

[0200] Example 2: Preparation of 3-(3,5-dimethyl-1H-pyrazol-4-yl)propan-1-ol (compound of formula V).

[0201] [ka]

[0202] The compound of formula IV (100 g, 632 mmol, 1.0 equiv) was dissolved in ethanol (1 L). To the solution was added hydrazine hydrate (87 g, 1738 mmol, 2.75 equiv) and concentrated HCl (4.6 mL, 0.2 equiv) at room temperature. The reaction mixture was heated to 75±5° C. and stirred at this temperature for 3 h. After completion of the reaction was confirmed by TLC (70% ethyl acetate:n-hexane, visible in iodine) and observation of the product peak in the mass spectrum, the reaction mixture was concentrated under reduced pressure to give the compound of formula V (70 g) as a colorless liquid syrup, which was used directly in the next step.

[0203] Example 3: Preparation of 4-(3-bromopropyl)-3,5-dimethyl-1H-pyrazole (compound of formula VIa)

[0204] [ka]

[0205] The compound of formula V (35 g, 227 mmol, 1.0 equiv.) was dissolved in 1,2-dichloroethane (525 mL). 3 (64.67 mL, 681 mmol, 3 equiv.) was added in small portions at room temperature over 30 min. The reaction mixture was heated to 75±5°C and stirred at this temperature for 3 h. After completion of the reaction as determined by TLC (50% ethyl acetate:n-hexane, visible in iodine) and observation of the product peak in the mass spectrum, the reaction mixture was diluted with dichloromethane (350 mL) and saturated NaHCO 3 The mixture was quenched with a solution of 0.1% MgSO 4 until pH = 7-8. The organic and aqueous phases were separated and both were collected. The organic phase was diluted with MgSO 4 4 The mixture was dried over water and filtered, and the filtrate was concentrated under reduced pressure to give the compound of formula VIa (38 g) as a viscous orange liquid.

[0206] Example 4: Preparation of 3-[3-(3,5-dimethyl-1H-pyrazol-4-yl)propoxy]-4-fluorobenzoic acid methyl ester (compound of formula VIIIa)

[0207] [ka]

[0208] The compound of formula VIIa (19 g, 111 mmol, 1.0 equiv.) was dissolved in DMF (190 mL). The compound of formula VIa (31.5 g, 145.14 mmol, 1.3 equiv.) was then added to K 2 CO 3 (38.6 g, 279.18 mmol, 2.5 equiv) was added at room temperature under stirring conditions. The reaction mixture was stirred at room temperature for 16-18 h. After completion of the reaction was confirmed by TLC (50% ethyl acetate: n-hexane), the reaction mixture was diluted with water (190 mL) and ethyl acetate (95 mL). The organic and aqueous phases were separated and both were collected. The aqueous phase was extracted with ethyl acetate (190 mL). The combined organic extracts were washed with water (95 mL), brine (95 mL) and diluted with Na 2 SO 4The mixture was dried over hexane and filtered. The filtered organic phase was concentrated under reduced pressure to give a crude viscous orange liquid (40 g). The crude was further purified by column chromatography on silica gel eluted with varying amounts of ethyl acetate in hexane to give the pure product, compound of formula VIIIa (25 g), as an off-white solid.

[0209] Example 5: Preparation of 3-[3-(3,5-dimethyl-1H-pyrazol-4-yl)propoxy]-4-fluorobenzoic acid methyl ester (compound of formula VIIIa)

[0210] [ka]

[0211] 4-(3-Bromopropyl)-3,5-dimethyl-1H-pyrazole hydrobromide (VIa) and DMSO were charged to a vessel and stirred at 20±10° C. for 10 minutes. The mixture was then heated to 55±5° C. with stirring. To this mixture was transferred a stirred solution containing 4-fluoro-3-hydroxybenzoic acid methyl ester (VIIa), potassium carbonate, and anhydrous DMSO. The DMSO solution of the alkyl bromide was charged slowly to maintain an internal temperature of 55.0±5° C. The addition was complete after 6 hours and the mixture was stirred for an additional hour at a temperature of 55.0±5° C. The mixture was cooled to 25±5° C. over 30 minutes and water was added while maintaining the temperature below 25° C. The mixture was extracted with ethyl acetate and the aqueous phase was back extracted with ethyl acetate. The combined ethyl acetate solution was washed with brine. The combined ethyl acetate washes were concentrated under reduced pressure to a minimum volume and heptane was added to precipitate VIIIa. The mixture was heated to 75±5° C. and aged with stirring for 1 hour. The mixture was cooled to 25±5° C. over 2 hours and the resulting solid was collected by filtration. The filter cake was washed with ethyl acetate (30%) in heptane. The isolated solid was dried under a stream of nitrogen. The solid was placed in a container and mixed with ethyl acetate and heptane. The resulting mixture was heated to 75±5° C. to dissolve the solid. The solution was cooled to 25±5° C. over 2 hours and the resulting solid was collected by filtration. The solid was washed with a 30% ethyl acetate / heptane solvent mixture and dried in a vacuum oven at 55° C. to give VIIIa of >99.5% purity.

[0212] Example 6: Preparation of 3-[3-(3,5-dimethyl-1H-pyrazol-4-yl)propoxy]-4-fluorobenzoic acid (compound of formula IX)

[0213] [ka]

[0214] The compound of formula VIIIa (19 g, 62 mmol, 1 equiv.) was dissolved in methanol (95 mL, 5 vol.) at room temperature. LiOH·H in water (57 mL) was added. 2A solution of 0 (6.5 g, 155 mmol, 2.5 equiv.) was added in portions over 10-15 min at room temperature. The reaction mixture was stirred at room temperature for 2 h. After completion of the reaction was confirmed by TLC (70% ethyl acetate:n-hexane), the reaction mixture was concentrated under reduced pressure below 45° C. to give a solid residue of formula IX.

[0215] Example 7: Preparation of Pharmacologically Acceptable Salts of Formula I The solid residue of formula IX was dissolved in water (57 mL), stirred for 10 min, and cooled to 0±5° C. The aqueous solution was acidified to pH=2 with concentrated HCl (20-25 mL) and stirred for 30 min at 0±5° C. A precipitate was observed, which was filtered and dried at room temperature to give the pure product compound of formula Ia (17.5 g) as an off-white solid.

[0216] Example 8: Preparation of additional pharma- ceutically acceptable salts of formula I

[0217] [ka]

[0218] Water and concentrated HCl were charged to a vessel and cooled to 10±5° C. with stirring. A compound of formula IX and water were placed in a second vessel and cooled to 10±5° C. with stirring. The HCl solution in vessel 1 was transferred into the vessel containing the mixture of compound of formula IX over a period of up to 15 minutes while maintaining the temperature at ≦25° C. The resulting slurry was aged with stirring at 20±5° C. for 44 hours. The solid was collected by filtration, washed with 0.2 N HCl (3×) and dried under vacuum at ≧55° C. to give Ia as a white solid in >99.8% purity.

[0219] Example 9: Preparation of 3-[3-(3,5-dimethyl-1H-pyrazol-4-yl)propoxy]-4-fluorobenzoic acid hydrochloride (compound Ia) from VIIIa

[0220] [ka]

[0221] A jacketed glass vessel was charged with the compound of formula VIIIa (1.0 equiv.) and methanol. The mixture was cooled to 10±5° C. with stirring and an aqueous solution of sodium hydroxide (3 equiv.) was added over 20 min. The mixture was aged with stirring at 20±5° C. for at least (NLT) 2 h, at which point the reaction was complete. Stirring was stopped and water was added. Methanol was then removed by vacuum distillation at an internal temperature not exceeding (NMT) 35° C. The resulting concentrated clear aqueous solution was cooled to 10° C. and concentrated HCl was added until the pH was reduced to between 1.4 and 1.6 (pH meter value) to precipitate the HCl salt. The solid was collected by filtration, washed with 0.2 N HCl, and dried under vacuum at 50° C. to give the compound of formula Ia in at least (NLT) 99.5% purity.

[0222] Example 10: Preparation of 3-[3-(3,5-dimethyl-1H-pyrazol-4-yl)propoxy]-4-fluorobenzoic acid (compound of formula IX) from VIIIa

[0223] [ka]

[0224] Methyl 3-[3-(3,5-dimethyl-1H-pyrazol-4-yl)propoxy]-4-fluorobenzoate (compound of formula VIIIa) and methanol were charged to a vessel and the resulting mixture was stirred at 20±5° C. until dissolved. The solution was cooled to 10±5° C. and sodium hydroxide solution was added over 20 minutes while maintaining the temperature at ≦25° C. The temperature of the mixture was adjusted to 25±5° C. and aged with stirring for 18 hours. The reaction mixture was then filtered. Water was added to the filtrate and the mixture was concentrated under reduced pressure until the mixture volume was at a minimum. Water was again added and the resulting mixture was concentrated under reduced pressure until the mixture volume was at a minimum. The pH of the aqueous mixture was adjusted to 5.5±0.5 by the addition of concentrated hydrochloric acid followed by 0.5 N HCl. The temperature of the mixture was adjusted to 7±5° C. and aged with stirring for an additional hour. The solid was collected by filtration, washed with water, and partially dried under vacuum at ≧55° C. to give the compound of formula IX as a white solid in >99.5% HPLC purity.

[0225] Example 11: Conversion of the hydrochloride salt to the free base 3-[3-(3,5-dimethyl-1H-pyrazol-4-yl)propoxy]-4-fluorobenzoic acid hydrochloride (10.0 g, 30.4 mmol, 1.0 equiv.) was taken in deionized water (30.0 mL) at room temperature and cooled to 10±5° C. Saturated sodium bicarbonate was added to the mixture until pH ≈ 6-7 and stirred at this temperature for 30 min. The resulting off-white precipitate was filtered and washed with deionized water (20 mL). The solid compound was dried at room temperature to give 3-[3-(3,5-dimethyl-1H-pyrazol-4-yl)propoxy]-4-fluorobenzoic acid (compound of formula IX) (7.40 g, 83.2%) as an off-white solid.

[0226] Example 12: Oral Administration Using Pharmaceutically Acceptable Salts of Formula I The following example describes the pharmacokinetic measurements of various salt and zwitterionic forms of the compound of formula IX. The results shown here demonstrate that the compound of formula I has a significantly enhanced pharmacokinetic profile.

[0227] Rats or dogs were orally administered the zwitterion, Na salt or HCl salt of AG-10. The form and dose of AG-10 used are as indicated in Table 3. Plasma samples from each rat / dog were measured between 0 and 96 hours after administration of the particular form of AG-10, as appropriate. After isolation from the animals, proteins were precipitated by adding 500 μL of 0.1% formamide in acetonitrile to each sample (50 μL). After addition of the formamide solution, the samples were vordex mixed and centrifuged at 1400 rpm for 15 minutes at 4°C. 100 μL of the supernatant was taken and diluted with 100 μL of water. 5 μL of the diluted sample was injected for LC-MS / MS analysis. Pharmacokinetic data were reported as shown in Table 3. max The values ​​and exposure dose (0-24 h, ng.h / mL) were used for calculations.

[0228] [Table 3]

[0229] As can be seen from Table 3, the HCl salt of Formula I exhibited a significantly lower C max Compare row 3 with row 1 and row 6 with row 4 of the table. Thus, to reach the same level of bioavailability, the HCl salt of Formula I requires a significantly lower dosage.

[0230] Example 13: Intravenous Administration of a Pharmaceutically Acceptable Salt of Formula I The following examples describe the pharmacokinetic measurements of various salt and zwitterionic forms of the compound of formula IX when administered intravenously to rats and dogs. The results presented here demonstrate that the compound of formula I has an unexpectedly high pharmacokinetic profile when administered orally and intravenously.

[0231] Mice, rats or dogs were administered AG-10 zwitterion, Na salt or HCl salt intravenously. The form and dose of AG-10 are shown in Table 4. Plasma samples from each mouse / rat / dog were measured at 0 and 24 hours after administration of the particular form of AG-10, as appropriate. After isolation from the animals, proteins were precipitated by adding 500 μL of 0.1% formamide in acetonitrile to each sample (50 μL). After addition of the formamide solution, the samples were vordex mixed and centrifuged at 1400 rpm for 15 minutes at 4°C. 100 μL of the supernatant was taken and diluted with 100 μL of water. 5 μL of the diluted sample was injected for LC-MS / MS analysis. Pharmacokinetic data were reported at the C-values ​​shown in Table 4. max The values ​​and exposure dose (0-24 h, ng.h / mL) were used for calculations.

[0232] [Table 4]

[0233] Example 14: High bioavailability of AG-10 in multiple species Figure 2 shows the pharmacokinetic results demonstrating the high bioavailability of AG-10 in dogs, rats and mice. Mean plasma concentrations of AG-10 were measured 0-24 hours after intravenous administration of 1 mg / kg AG-10 and oral administration of 5 mg / kg AG-10. The calculated pharmacokinetic data are shown in Figure 2.

[0234] Example 15: High bioavailability of AG-10 in dogs Figure 3 shows the pharmacokinetic results demonstrating the high bioavailability of AG-10 in male / female dogs at different dose levels. Mean plasma concentrations of AG-10 were measured 2-24 hours after oral administration of 5 mg / kg or 20 mg / kg of AG-10. The calculated pharmacokinetic data are shown in Figure 3.

[0235] Example 16: Salt and co-crystal screening A number of salts and co-crystal forms were tested for various targeted pharma- ceutically acceptable salts. Experimental details can be found in Tables 5 and 6. Experiments were performed using various crystallization techniques such as cooling, evaporation, slurrying, and grinding with solvents. Solids obtained from salt and co-crystal screening experiments were observed by polarized optical microscopy (PLM) and analyzed by XRPD. XRPD patterns of isolated solids were compared to those of known forms of AG-10 and counterions / coformers.

[0236] The confirmed salts of AG10 were identified from experiments targeting salt formation with strong acids, specifically methanesulfonic acid, ethane-1,2-disulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, ethanesulfonic acid, sulfuric acid, hydrogen bromide, and nitric acid.

[0237] Salts or co-crystals of AG10 were also isolated from experiments targeting salt / co-crystal formation with weak organic acids such as citric acid, acetic acid, maleic acid, oxalic acid and malic acid.

[0238] Experiments targeting L-tartaric, glycolic and fumaric acids were also designed; however, experiments performed aimed at isolating these materials as single crystalline phases were unsuccessful and produced physical mixtures of the starting material or the starting material with unique additional peaks.

[0239] Attempts to produce salts / co-crystals of AG10 with L-aspartic acid, benzoic acid, gentisic acid and succinic acid were unsuccessful.

[0240] [Table 5]

[0241] [Table 6]

[0242] [Table 7]

[0243] [Table 8]

[0244] [Table 9]

[0245] [Table 10]

[0246] Example 17: Preparation of the mesylate salt of formula IX The mesylate salt of formula IX was prepared by adding 1 molar equivalent of methanesulfonic acid to a solution of AG-10 in MEK:DMF=2:0.3 (v / v) at elevated temperature. The suspension was kept at elevated temperature for about 20 minutes, cooled to room temperature and the solid was isolated.

[0247] The XRPD pattern is shown in Figure 4 and shows the separation of the peaks indicative of a crystalline material. Indexing of the XRPD pattern was attempted, however no indexing solution was found, possibly due to the sample containing a mixture of crystalline phases or poor peak resolution.

[0248] 1 The H NMR spectrum was consistent with AG-10 mesylate at a 1:1 molar ratio based on the peak at 2.37 ppm. Traces of DMF and additional unknown peaks were also observed in the spectrum.

[0249] The DSC thermogram (Figure 5) shows a single endotherm at approximately 233°C (peak maximum) that can be attributed to melting. No significant weight loss is observed in the TGA (Figure 5) up to approximately 200°C, suggesting that the material is likely unsolvated / anhydrous.

[0250] Example 18: Preparation of the edisylate salt of formula IX The edisylate salt of formula IX was prepared by adding one molar equivalent of 1,2-ethanedisulfonic acid to a solution of AG10 in acetone:DMA at elevated temperature. The suspension was cooled to ambient temperature and the solid was isolated.

[0251] By XRPD, the edisylate salt of formula IX consists of crystalline material (Figure 6). 1 The H NMR spectrum is consistent with a 2:1 molar ratio of AG-10 edisylate based on the peak at 2.7 ppm. Approximately 1 molar of DMA was also observed, suggesting an AG-10 edisylate DMA (2:1:1) solvate.

[0252] The DSC thermogram (Figure 7) shows a broad feature at ~139 °C associated with a weight loss of 11% based on the TGA (Figure 7) data, likely due to desolvation. A sharp endotherm is observed at 313 °C (peak maximum) likely due to melting / decomposition of the desolvated material. To further understand the behavior of the material upon heating, hot stage microscopy is recommended.

[0253] A sample of the edisylate salt of Formula IX was dried at 180° C. for 10 minutes, which did not result in any change in physical form based on XRPD.

[0254] Example 19: Preparation of the besylate salt of formula IX The besylate salt of Formula IX was prepared from cooling a THF solution containing equimolar equivalents of AG-10 and benzenesulfonic acid.

[0255] The besylate salt of formula IX consists of crystalline material and the XRPD pattern is shown in FIG. 1 The H NMR spectrum is generally consistent with AG-10 besylate salt in approximately a 1:1 ratio. Traces of THF were also observed in the spectrum referenced to a peak at 3.6 ppm.

[0256] Two endotherms are observed in the DSC thermogram with peak maxima at approximately 158° C. and 177° C. (FIG. 9). A weight loss of 0.2% is observed between 42° C. and 127° C. (FIG. 9).

[0257] Example 20: Preparation of the tosylate salt of formula IX The tosylate salt of formula IX was prepared by adding 1 molar equivalent of p-toluenesulfonic acid to a solution of AG-10 in acetonitrile at elevated temperature.

[0258] By XRPD, the tosylate salt of formula IX is composed of crystalline material (Figure 10). The pattern was successfully indexed, indicating that the material consists primarily or exclusively of a single crystalline phase. The unit cell volume obtained from the indexing solution is consistent with the AG10 tosylate (1:1) salt, taking into account the molecular volumes.

[0259] 1 The 1 H NMR spectrum is generally consistent with AG10 tosylate salt in about a 1:1 molar ratio based on the peak at 2.28 ppm.

[0260] The DSC thermogram shows a single endotherm at about 205° C. (peak maximum) that is likely due to melting (FIG. 11). No significant weight loss was observed in the TGA until about 160° C., suggesting that the material is likely unsolvated / anhydrous (FIG. 11).

[0261] Example 21: Preparation of the esylate salt of formula IX The esylate salt of formula IX was precipitated from a THF solution containing AG-10 and ethanesulfonic acid (1:1 molar ratio) at 50° C. The suspension was cooled and the solid was isolated.

[0262] The esylate salt of formula IX consists of crystalline material as confirmed by XRPD (Figure 12). 1 The H NMR spectrum is consistent with a 1:1 molar ratio of AG-10 esylate salt based on the peak at 2.4 ppm. Approximately 0.1 moles of THF was observed in the spectrum.

[0263] In the DSC thermogram, one endotherm is observed at 199° C. (peak maximum), likely due to melting (FIG. 13). There is no significant weight loss upon heating to melting, suggesting that the material is unsolvated or anhydrous (FIG. 13).

[0264] Example 22: Preparation of the bromide salt of formula IX The bromide salt of formula IX was prepared by adding an equimolar amount of hydrogen bromide to a solution of AG-10 in MIBK:DMSO 2:0.4 (v / v) at ~60°C. This resulted in the formation of a yellow solution and oil. The sample was placed in a vacuum oven at room temperature for 3 days, resulting in an oil with some solids present. MEK was added to the sample with sonication, heated to 60°C, and then cooled twice. The solids remaining in the resulting suspension were isolated and analyzed.

[0265] The bromide salt of formula IX consists of crystalline material (Figure 14). 1 The H NMR spectrum is consistent with the chemical structure of AG-10. Approximately 1 molar DMSO was also observed, based on the peak at 2.54 ppm.

[0266] The bromide content was found to be 17.7% by weight based on IC, which is consistent with the calculated bromide content of the AG-10 bromide DMSO (1:1:1) solvate (17.7%).

[0267] An endotherm at -105°C (peak maximum) followed by an exotherm at 155°C (peak maximum) and an endotherm at -214°C are observed in the DSC data (Figure 15). Upon heating to approximately 182°C, a weight loss of 19.9% ​​is observed, possibly related to solvent loss and possible recrystallization to the unsolvated form (Figure 15).

[0268] Example 23: Preparation of the nitrate salt of formula IX Two nitrate forms of formula IX have been identified, designated form a and form b. Nitrate, a-form Form a of the nitrate salt of Formula IX was precipitated from a DMSO solution containing an equimolar ratio of AG-10 and nitric acid. The AG-10 nitrate material A consists of a single crystalline phase based on successful indexing of the XRPD pattern (FIG. 16).

[0269] AG10 Nitrate Form A Solution 1 The H NMR spectrum is consistent with the chemical structure of AG-10. Approximately 0.8 moles of DMSO are present based on the peak at 2.54 ppm. Water and slight additional peaks are also observed.

[0270] The DSC thermogram shows a broad endotherm around 117° C., which is associated with a 2.5% weight loss likely due to loss of volatiles (FIG. 17). This broad endotherm is followed by an exotherm with a peak maximum at ∼173° C., which is associated with a ∼16% weight loss likely due to melting / decomposition (FIG. 17).

[0271] The nitrate content was found to be 7.5 wt.% based on IC, a value that does not match the calculated nitrate content predicted for the unsolvated 1:1 nitrate (theoretical nitrate content: 17.5%) or even the calculated nitrate content predicted for the 1:1:1 AG-10 nitrate DMSO solvate (theoretical nitrate content: 14.3%).

[0272] Nitrate, b-type Form b of the nitrate salt of formula IX was prepared by evaporating a THF solution containing equimolar ratios of AG-10 and nitric acid, and the XRPD pattern of this solid is shown in Figure 18. AG-10 Solution of Nitrate Substance B 1 The 1 H NMR spectrum is consistent with the chemical structure of AG-10.

[0273] The nitrate content was found to be 16.9 wt % based on IC, which is generally consistent with AG-10 nitrate at about 1:1.

[0274] Example 24: Preparation of the sulfate salt of formula IX The sulfate salt of formula IX is prepared by evaporating an ethanol solution containing equimolar amounts of AG-10 and sulfuric acid and cooling (from 60° C. to 2-8° C.) The sulfate salt of formula IX consists of a crystalline material (FIG. 19).

[0275] 1 The H NMR spectrum confirmed the presence of AG-10 and showed that it contained approximately 1 mole of ethanol based on the peaks at 1.06 ppm and 3.4 ppm. Additional unknown peaks were also observed in the spectrum.

[0276] The sulfate content was found to be 15.9 wt % based on IC, which corresponds to an AG-10:sulfate ratio of 1:0.58. A 6.4% weight loss was observed in the TGA thermogram between ~30°C and 96°C, equivalent to 1 mole of ethanol, estimated to be the AG10 sulfate 2:1 salt (Figure 20). A broad feature was observed by DSC (Figure 20).

[0277] Example 25: Preparation of the citrate salt of formula IX After evaporation at room temperature of the IPA solution saturated with citric acid and containing AG-10, a single crystal of the citrate salt of formula IX was obtained. After recovery of a single crystal suitable for SCXRD, the sample was further evaporated and the collected solid consisted of a mixture of AG-10 citrate salt and citric acid based on XRPD. The structure of AG-10 citrate has been successfully determined. The crystal system is triclinic and the space group is

[0278]

number

[0279] The unit cell parameters and the calculation volume are

[0280] [ka]

[0281] The formula weight is 484.43 g mol -1 and Z = 2, resulting in a calculated density of 1.435 g cm -3 It is.

[0282] A second experiment was performed with the aim of obtaining a bulk solid of AG10 citrate as a single crystalline phase for further characterization, which also resulted in a physical mixture of AG10 citrate and citric acid.

[0283] Example 26: Preparation of the oxalate salt of formula IX The oxalate salt of formula IX was precipitated from a DMA solution containing AG-10 and oxalic acid (1:1 molar ratio) at 50° C. The sample was cooled to room temperature and the solid was isolated for characterization.

[0284] By XRPD, the oxalate salt of Formula IX consists of crystalline material (Figure 21). The XRPD pattern of this sample was successfully indexed, indicating that the sample consists primarily or exclusively of a single crystalline phase. The indexed volume is consistent with the AG-10 hemi-oxalate salt based on molecular volume considerations.

[0285] 1 The H NMR spectrum is consistent with the chemical structure of AG-10. Approximately 0.1 molar DMA and water were also present in the spectrum.

[0286] The oxalate content of the sample was determined by IC to be 13.7%, confirming the approximately 2:1 stoichiometry of AG-10 hemi-oxalate.

[0287] A single endotherm is observed in the DSC data at ~225 °C (peak maximum) that is likely due to melting / decomposition based on the TGA data (Figure 22). The TGA thermogram likely indicates that 1 It shows an initial weight loss of 0.9% upon heating between 33 °C and 169 °C, due to loss of residual surface solvent as observed by DMA by H NMR (Figure 22).

[0288] Example 27: Preparation of the maleate salt of formula IX Two maleate forms of formula IX have been identified, designated form a and form b. Maleate, a-form Addition of a solution of maleic acid in nitromethane (2.2 molar equivalents) to AG-10 at 70° C. produced a suspension, which was cooled to room temperature and reheated to 60° C. twice before isolating the solid.

[0289] Form a of the maleate salt of formula IX consists of crystalline material based on XRPD (Figure 23). The XRPD pattern cannot be indexed, suggesting that the material is not composed of a single crystalline phase and is a mixture of possible forms. XRPD analysis suggests that form a of the maleate salt of formula IX was isolated as a mixture with form b of the maleate salt of formula IX.

[0290] Sample 1 The H NMR spectrum contained approximately a 1:1 molar ratio of AG-10:maleic acid based on the peak at 6.23 ppm. Approximately 1.3 moles of nitromethane are observed for every mole of AG-10 based on the presence of a peak at 4.42 ppm. Additional unknown minor peaks were also observed in the spectrum.

[0291] An endotherm at .about.160.degree. C. (peak maximum) is observed in the DSC data (FIG. 24). Upon heating to 110.degree. C., a weight loss of 8.4% is observed, likely due to loss of solvent (FIG. 24). A sample of form a of the maleate salt of Formula IX was dried at 110.degree. C. for approximately 7 minutes to yield a disordered material with peaks consistent with form b of the maleate salt of Formula IX based on XRPD.

[0292] The sample was analyzed by XRPD and 1 Based on 1 H NMR data, it appears to consist of a mixture of Form b of the maleate salt of Formula IX, as well as a possible nitromethane solvate.

[0293] Maleate, b-form Form b of the maleate salt of formula IX was prepared from a temperature-elevated slurry experiment containing AG-10 and maleic acid (1:1) in p-dioxane. The XRPD pattern of form b of the maleate salt of formula IX (Figure 25) was successfully indexed, indicating that the material consisted primarily or exclusively of a single crystalline phase. The indexed amounts were consistent with a 1:1 AG-10 maleate salt.

[0294] Sample 1 The H NMR spectrum is consistent with a 1:1 molar ratio of AG-10 and maleic acid. Approximately 0.3 moles of p-dioxane was also observed in the spectrum.

[0295] A single endotherm is observed in the DSC thermogram at approximately 171° C. (peak maximum) (FIG. 26). No significant weight loss is observed upon heating the sample between 33° C. and 120° C. (FIG. 26).

[0296] Example 28: Preparation of acetate salt of formula IX The acetate salt of formula IX was prepared by directly grinding AG-10 with acetic acid in a 1:1 molar ratio.

[0297] By XRPD, the acetate salt of Formula IX was composed of crystalline material and is shown in Figure 27. The XRPD pattern was successfully indexed, indicating that the sample consisted primarily or exclusively of a single crystalline phase. 1 The 1 H NMR spectrum is consistent with the chemical structure of AG-10, with approximately 0.9 moles of acetic acid present.

[0298] The DSC thermogram showed a broad endothermic peak at ~113 °C associated with a ~16% weight loss that could be attributed to the loss of acetic acid (Figure 28). This was followed by endothermic peaks at 186 °C and 192 °C (peak maximum) likely due to the melting of the free form of AG-10 (Figure 28).

[0299] Example 29: Preparation of the L-malate salt of formula IX A saturated solution of L-malic acid in nitromethane was added to AG-10 at 60° C., and the resulting solution was cooled to produce a solid at subambient temperatures. The XRPD pattern consisted of a unique crystalline material designated as the L-malate salt of formula IX ( FIG. 29 ).

[0300] 1 The 1 H NMR spectrum contained 1.8 moles of malic acid per mole of AG-10 based on the peak at 4.2 ppm. Small amounts of ACN and water were also observed in the spectrum.

[0301] Two broad endotherms are observed in the DSC data with peak maxima at 89° C. and 199° C. (FIG. 30). A weight loss of 0.2% is observed between 33° C. and 107° C. (FIG. 30).

[0302] Example 30: Crystalline Form A of Formula IX The material of formula Ia (HCl salt of formula IX) prepared in Example 7 was characterized by X-ray powder diffraction (XRPD) (Figure 31), thermogravimetric analysis (TGA) (Figures 32-34), differential scanning calorimetry (DSC) (Figures 32-34) and polarized light microscopy (PLM) (Figure 35). This material was designated as crystalline Form A of formula IX. Three different XRPD plots representing three different preparations obtained according to Example 7 are overlaid in Figure 31. Representative peak values ​​for the XRPD plot shown in Figure 31 are provided in Table 7 below.

[0303] [Table 11]

[0304] Three individual TGA / DSC plots of crystalline Form A of Formula IX are shown in Figures 32-35. Thermogravimetric analysis measured a weight loss of about 0.7%-1.9% when heated near about 150°C, and further characterization using differential scanning calorimetry shows at least two endothermic peaks at about 211-214°C and 237-239°C. The HPLC purity of crystalline Form A of Formula IX was determined to be 98.76 area %.

[0305] The asymmetric unit of crystalline Form A of Formula IX is shown in Figure 36. It contains one cation of compound AG10 free base and one chloride ion (an HCl molecule has transferred a proton to the N1 atom of the free base), indicating that Form A is in the form of anhydrous mono-HCl salt.

[0306] To evaluate the hygroscopicity and physical stability of crystalline Form A of Formula IX under different humidities, Dynamic Vapor Sorption (DVS) data was collected at 25° C. after pre-equilibrating the sample at 0% RH to remove unbound water (free water). The DVS results (FIG. 37) showed a water absorption rate of 1.6% at 25° C. / 80% RH, suggesting that crystalline Form A of Formula IX is slightly hygroscopic. Furthermore, the XRPD results (FIG. 38) showed no morphological changes before and after the DVS study.

[0307] Example 31: Polymorphism screening of AG-10 Using crystalline form A of formula IX as the starting material, polymorph screening experiments were performed under 98 conditions by vapor diffusion, antisolvent addition, slurry conversion, slow evaporation, and slow cooling methods. A total of 10 additional crystalline forms were obtained from polymorph screening and follow-up studies, including six HCl salt forms (forms A / B / E / H / I / J), two free base forms (forms C / G), and two currently unidentified forms (forms D / F). The crystalline forms A / B / E were identified as anhydrous. Form I was identified as a hydrate. Forms H and J were identified as MeOH and DMAc solvates, respectively. The methods used and the crystalline forms identified are summarized in Table 8.

[0308] [Table 12]

[0309] Antisolvent addition A total of 24 anti-solvent addition experiments were performed. For each experiment, approximately 15 mg of crystalline form A of formula IX was weighed into a 20 mL glass vial, followed by the addition of 0.125-0.63 mL of the corresponding solvent. The mixture was then magnetically stirred at a speed of 750 RPM at room temperature to obtain a clear solution. The corresponding anti-solvent was then added to the solution to induce precipitation or until the total amount of anti-solvent reached 10.0 mL. The clear solution was slurried at 5° C. If no precipitation occurred, the solution was transferred to fast evaporation at RT or vacuum drying at RT. The solid was isolated for XRPD analysis. The results summarized in Table 9 show that Form A, Form C, Form D, Form E, and Form A with additional peaks were obtained.

[0310] [Table 13]

[0311] Slow Evaporation Slow evaporation experiments were carried out under seven conditions. For each experiment, approximately 15 mg of crystalline form A of formula IX was weighed into a 3 mL glass vial, followed by addition of the corresponding solvent or solvent mixture to obtain a clear solution. The vial was then covered with parafilm with 3-4 pin holes and the solution was kept at room temperature to allow the solution to slowly evaporate. The isolated solid was examined by XRPD. Forms A and H were produced, as summarized in Table 10.

[0312] [Table 14]

[0313] Slow cooling Slow cooling experiments were carried out with eight solvent systems. For each experiment, about 15-35 mg of crystalline form A of formula IX was suspended in 0.8-2.0 mL of the corresponding solvent in a 3 mL glass vial at room temperature. The suspension was slurried at 50° C. with a magnetic stirrer at a speed of 750 RPM. The sample was equilibrated at 50° C. for 1 hour and filtered through a 0.45 μm PTFE membrane. The filtrate was then slowly cooled from 50° C. to 5° C. at a rate of 0.1° C. / min. If no precipitation occurred, the solution was transferred to fast evaporation at room temperature or vacuum drying at room temperature. The results summarized in Table 11 show that Forms A, E and J were obtained.

[0314] [Table 15]

[0315] Slurry conversion at RT Slurry conversion experiments were carried out at room temperature in different solvent systems. For each experiment, approximately 15-35 mg of crystalline form A of formula IX was suspended in 0.3-2.0 mL of the corresponding solvent in a 1.5 mL glass vial. The suspension was magnetically stirred at room temperature for 4 days, after which the remaining solid was isolated for XRPD analysis. The results summarized in Table 12 show that Forms A, C and G were obtained.

[0316] [Table 16]

[0317] Slurry conversion at 50°C Slurry conversion experiments were carried out in different solvent systems at 50° C. For each experiment, approximately 15 mg of crystalline form A of formula IX was suspended in 1.0 mL of the corresponding solvent in a 1.5 mL glass vial. The suspension was magnetically stirred at 50° C. for 4 days, after which the remaining solid was isolated for XRPD analysis. The results summarized in Table 13 show that Forms A and F were obtained.

[0318] [Table 17]

[0319] Solid-phase vapor diffusion Solid-phase vapor diffusion experiments were carried out with 13 solvents. For each experiment, approximately 15 mg of crystalline form A of formula IX was weighed into a 3 mL vial and placed in a 20 mL vial with 4 mL of the corresponding solvent. The 20 mL vial was sealed with a cap and kept at room temperature for 39 days to allow the solvent vapor to interact with the solid sample. The isolated solid was examined by XRPD. The results summarized in Table 14 show that crystalline form A and form A with an extra peak were obtained.

[0320] [Table 18]

[0321] Liquid-phase vapor diffusion 21 liquid phase vapor diffusion experiments were performed. For each experiment, approximately 15 mg of crystalline form A of formula IX was dissolved in 0.125-0.6 mL of the corresponding solvent in a 3 mL vial to obtain a clear solution. The solution was then placed in a 20 mL vial along with 4 mL of the corresponding anti-solvent. The 20 mL vial was sealed with a cap and kept at room temperature to allow sufficient time for the solvent vapor to interact with the solution. If no precipitate formed, the solution was then transferred to a flash evaporation at room temperature. The solid was isolated for XRPD analysis. The results summarized in Table 15 indicate that Form A was obtained.

[0322] [Table 19]

[0323] A chart summarizing the interchangeability between the identified crystal forms is shown in FIG. Solvent Abbreviations

[0324] [Table 20]

[0325] Apparatus and method XRPD A PANalytical X-ray powder diffractometer was used for XRPD analysis. The XRPD parameters used are listed in Table 17.

[0326] [Table 21]

[0327] TGA and DSC TGA data was collected using a TA Q500 / Q5000 TGA from TA Instruments. DSC was performed using a TA Q200 / Q2000 DSC from TA Instruments. The detailed parameters used are listed in Table 18.

[0328] [Table 22]

[0329] 1 H NMR 1 H NMR data is for DMSO-d 6 The NMR spectra were collected on a Bruker 400M NMR spectrometer using a NMR spectrometry.

[0330] DVS DVS was measured by DVS Intrinsic from SMS (Surface Measurement Systems, Inc.). The parameters for the DVS test are listed in Table 19.

[0331] [Table 23]

[0332] HPLC An Agilent 1100 / 1260 HPLC was used to analyze purity and solubility, detailed methods are listed in Table 20.

[0333] [Table 24]

[0334] I C Cl - The IC methods for content determination are listed in Table 21.

[0335] [Table 25]

[0336] Example 32: Preparation of Crystalline Form B of Formula IX Crystalline form B of formula IX was obtained by heating a sample of crystalline form A to 212°C, cooling to 30°C under nitrogen protection and exposing to air conditioning. The HPLC purity and stoichiometry (acid:free base (FB)) of crystalline form B were determined to be 97.86 area% and 0.86, respectively. The XRPD pattern is shown in Figure 40 and the TGA / DSC curve is shown in Figure 41. The results showed that form B was crystalline with a 1.2% weight loss before 150°C in TGA and three endothermic peaks at 161.4, 232.2 and 262.3°C (peaks) in DSC. Due to the limited TGA weight loss before 150°C and neat DSC, form B was assumed to be anhydrous. Heating experiments were performed to investigate the thermal signals. As shown in FIG. 42, after heating to 100° C. or 170° C., cooling to 30° C. under nitrogen protection, and then exposure to air, Form B transformed into Form I (Form I is described in more detail in Example 38). The peak values ​​of the XRPD plot shown in Figure 40 are provided in Table 22 below.

[0337] [Table 26]

[0338] Example 33: Preparation of Crystalline Form C of Formula IX Crystalline Form C of formula IX can be dissolved in DMSO / H at room temperature. 2 The XRPD was obtained by anti-solvent addition in 200 and is shown in Figure 43. The TGA and DSC results shown in Figure 44 showed a 3.1% weight loss up to 150°C and two endothermic peaks at 91.2°C and 173.0°C. - The content was 0.17% (theoretical Cl for mono-HCl salt). - The C form was confirmed to be the free base form, since the C form contained 10.8% of the active ingredient. The peak values ​​of the XRPD plot shown in Figure 43 are provided in Table 23 below.

[0339] [Table 27]

[0340] Example 34: Preparation of crystalline forms D and F of formula IX Crystalline form Form D of Formula IX was obtained by adding anti-solvent to MeOH / IPAc system at room temperature. Form F of Formula IX was obtained via slurrying Form A in toluene at 50° C. Their XRPD patterns are shown in Figure 45 (Form D, top plot; Form F, bottom plot). The peak values ​​for the XRPD plots (Forms D and F) shown in Figure 45 are provided in Tables 24 and 25 below.

[0341] [Table 28]

[0342] [Table 29]

[0343] Example 35: Preparation of crystalline form E of formula IX Crystalline Form E of formula IX can be dissolved in CHCl at room temperature. 3 The crystalline form E of formula IX was obtained by slow evaporation in 1000 mL / EtOH. The HPLC purity and stoichiometry (acid:FB) of crystalline form E of formula IX were determined to be 98.60 area % and 0.91, respectively. The XRPD pattern is shown in FIG. 46 and the TGA / DSC curve is depicted in FIG. 47. The results showed that crystalline form E had a 1.5% weight loss by 130° C. in TGA and two endothermic peaks at 182.0° C. and 242.7° C. in DSC (peaks). Due to the limited TGA weight loss and neat (smooth) DSC before 170° C., form E was assumed to be anhydrous. A heating experiment was performed to investigate the thermal signal at 182.0° C. (peak) in DSC. As shown in FIG. 48, form E was converted to the hydrate form, form I, after heating to 195° C. and cooling to 30° C. under the protection of nitrogen, followed by exposure to air. Based on the thermal data and heating experiments, the anhydrous Form E was transformed into a novel anhydrous form (the endothermic signal in DSC at ∼180°C could be a form transition signal), which then transformed into the hydrated form Form I upon interaction with moisture upon exposure to ambient conditions. The peak values ​​for the XRPD plot shown in Figure 46 are provided in Table 26 below.

[0344] [Table 30]

[0345] Example 36: Preparation of crystalline form G of formula IX Form G of formula IX can be dissolved in DMAc / H at room temperature. 2 The XRPD was obtained via slurry in 200° C. (v:v, 1:3) and is shown in FIG. 49. The TGA and DSC results shown in FIG. 50 showed a 3.7% weight loss up to 200° C. and one sharp endothermic signal at 231.1° C. (peak). - The content was 0.14% (theoretical Cl of mono-HCl salt). - The G form was confirmed to be in the free base form, since the G form contained 10.8% of the active ingredient. The peak values ​​for the XRPD plot shown in Figure 49 are provided in Table 27 below.

[0346] [Table 31]

[0347] [Table 32]

[0348] Example 37: Preparation of crystalline form H of formula IX Crystalline form Form H of formula IX was obtained by slow evaporation in acetone / MeOH system at RT and its XRPD is shown in Figure 51. The HPLC purity and stoichiometry (acid:FB) of Form H (810119-11-A4) were determined to be 98.47 area % and 0.91, respectively. The TGA and DSC curves (Figure 52) showed a weight loss of 4.6% before 120°C and three endothermic peaks at 90.4, 200.5 and 232.3°C (peaks). 1As shown in the H NMR spectrum (Figure 53), 0.36 equivalents of MeOH (~3.40 wt%) were detected. Combined with the fact that after heating Form H to 120°C under the protection of nitrogen, cooling to 30°C, and exposure to ambient conditions, a crystal form change to Form I was observed (Figure 54), leading to the prediction that Form H is a MeOH solvate. The peak values ​​of the XRPD plot shown in Figure 51 are provided in Table 28 below.

[0349] [Table 33]

[0350] Example 38: Preparation of Crystalline Form I of Formula IX Crystalline Form I of formula IX was obtained by heating crystalline Form B of formula IX to 100° C., cooling to 30° C. under the protection of nitrogen, and then exposing to air. Its XRPD is shown in FIG. 55. The HPLC purity and stoichiometry (acid:FB) of crystalline Form I of formula IX were determined to be 97.94 area % and 0.86, respectively. Crystalline Form I was obtained via a solid-state transition of anhydrous Form B, and a weight loss of 3.0% (equivalent to 0.5 moles of water) was observed with an endothermic peak at 62.0° C. (Peak, FIG. 56), so Form I was presumed to be a hydrate. The peak values ​​of the XRPD plot shown in Figure 55 are provided in Table 29 below.

[0351] [Table 34]

[0352] [Table 35]

[0353] To further identify the crystal form I and to investigate its dehydration behavior, 2In-situ XRPD analysis was performed using N flow to observe the dehydrated form of Form I, and a KF (Karl Fischer) test was performed to confirm whether the TGA weight loss was caused by moisture. 2 For Form I with purging (30°C / 16% RH), a morphological change to anhydrous Form B was observed. As shown in Figure 58, Form I showed a weight loss of 2.6% up to 120°C. Based on the KF results, a moisture content of about 3.48% was observed in the Form I sample. 2 Combined with its morphology change to anhydrous Form B under flow, Form I was identified as a hydrate.

[0354] Example 39: Preparation of Crystalline Form J of Formula IX Crystalline form J of formula IX was obtained by slow evaporation in MEK / DMAc system followed by vacuum drying at 50° C. Its XRPD is shown in FIG. 59. The HPLC purity and stoichiometry (acid:FB) of crystalline form J of formula IX were determined to be 91.69 area % and 0.90, respectively. The TGA and DSC results in FIG. 60 showed a weight loss of 21.5% up to 120° C. and three endothermic peaks at 120.8, 197.8 and 221.5° C. (peaks). 1 As shown in the H NMR spectrum (Figure 61), 4.9 equivalents of DMAc (~56.51 wt%) were detected. Combined with the fact that after heating Form J to 130 °C, cooling to 30 °C under nitrogen protection, and exposure to ambient conditions, a morphology change (highlighted) to a mixture of Form I and Form A was observed (Figure 62), leading to the speculation that Form J is a DMAc solvate.

[0355] The peak values ​​of the XRPD plot shown in Figure 59 are provided in Table 30 below. [Table 36]

[0356] [Table 37]

[0357] Example 40: Preparation of Crystalline Form K of Formula IX The free base material prepared in Example 11 was characterized by XRPD (Figure 63), TGA (Figure 64) and DSC (Figure 64). This material was designated as crystalline Form K of Formula IX. A 6.1% weight loss was observed in TGA up to 150°C, and DSC results showed endothermic peaks at 159.3, 176.2 and 278.4°C (peaks). The HPLC purity of crystalline Form K of Formula IX was determined to be 99.12 area %. The peak values ​​of the XRPD plot shown in Figure 63 are provided in Table 31 below.

[0358] [Table 38]

[0359] Although the above has been described in some detail by way of illustration and example for purposes of clarity and understanding, those skilled in the art will appreciate that certain changes and modifications can be practiced within the scope of the appended claims. Additionally, each reference provided herein is incorporated by reference in its entirety to the same extent as if each reference was individually incorporated by reference.

Claims

1. A crystalline form of 3-(3-(3,5-dimethyl-1H-pyrazol-4-yl)propoxy)-4-fluorobenzoic acid characterized by a powder X-ray diffraction pattern with peaks at 13.1, 15.1, 17.1, 17.8, and 24.7 degrees 2θ (±0.2 degrees 2θ).

2. ０１９１ The crystalline form of claim 1 further having peaks at one or more of 12.2, 13.4, 21.7, 24.3, 24.9, 26.6, and 28.3°2θ (±0.2°2θ).

3. The crystalline form of claim 1 further having peaks at two or more of 12.2, 13.4, 21.7, 24.3, 24.9, 26.6, and 28.3°2θ (±0.2°2θ).

4. The crystalline form of claim 1 further having peaks at three or more of 12.2, 13.4, 21.7, 24.3, 24.9, 26.6, and 28.3°2θ (±0.2°2θ).

5. The crystalline form of claim 1 further having peaks at four or more of 12.2, 13.4, 21.7, 24.3, 24.9, 26.6, and 28.3°2θ (±0.2°2θ).

6. The crystalline form of claim 1 further having peaks at 12.2, 13.4, 21.7, 24.3, 24.9, 26.6, and 28.3°2θ (±0.2°2θ).

7. 2. The crystalline form of claim 1, characterized by a powder X-ray diffraction pattern consistent with Figure 49 below. 【Chemistry 1】

8. A composition comprising the crystalline form of any one of claims 1 to 7.

9. 9. The composition of claim 8, comprising no more than 10% of another crystalline or amorphous form of 3-(3-(3,5-dimethyl-1H-pyrazol-4-yl)propoxy)-4-fluorobenzoic acid or its salt.

10. A pharmaceutical composition for treating a transthyretin (TTR) amyloid-related disease, comprising the crystalline form of any one of claims 1 to 7 or the composition of claim 8 or 9.

11. A pharmaceutical composition for treating transthyretin (TTR) amyloid cardiomyopathy, comprising the crystalline form of any one of claims 1 to 7 or the composition of claim 8 or 9.

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