OA17658A - Crystalline polymorphs of the free base of 2hydroxy-6-((2-(1-isopropyl-1H-pyrazol-5-yl)pyridin3-yl)methoxy)benzaldehyde. - Google Patents

Abstract

Disclosed are crystalline free base ansolvate forms of 2-hydroxy-6-((2-(1-isopropyl-lH-pyrazol5-yl)pyridin-3-yl)methoxy)benzaldehyde (or compound 1), such as the free base form I, form II and material N. Also disclosed are crystalline free base solvates of 2-hydroxy-6-((2-(1-isopropyl-1H-pyrazol-5-yl)pyridin-3-yl)methoxy)benzaldehyde (or compound 1).

Description

that is stable, is manufactured reproducibly, and has physicochemical properties favorable for its use as a therapeutic agent.

[0005] However, the art remains unable to predict which crystalline form of an agent will hâve a combination of the desired properties and will be suitable for human administration, and how to make the agent in such a crystalline form.

Summary

Ansolvates [0006] This invention arises in part out the discovery that an HCl sait of Compound 1 disproportionates or loses HCl, and a disproportionation of the HCl sait of Compound 1 in water generates the free base and disproportionation was facile upon exposure to elevated humidity, with wet milling, and in direct contact with water (e.g. slurry). The sulfate sait of Compound 1 also disproportionates from certain solvents such as dimethyl sulfoxide and methanol when precipitated with water. The volatilization of HCl was évident within hours of exposure to drying conditions. For example, partial conversion to the free base was observed within 12 hours at 30 °C. Accordingly, the free base of Compound 1 provides a stabler chemical entity compared to the corresponding HCl or sulfate and such other sait.

[0007] It has now been discovered that 2-hydroxy-6-((2-(l-isopropyl-lH-pyrazol-5-yl)pyridin-

3-yl)methoxy)benzaldehyde (or Compound 1) i.e., the free base of Compound 1, can be obtained as one or more crystalline ansolvate forms, several of which are referred to here as crystalline Form I, Form II and Material N. In preferred embodiments, the free base of Compound 1 is a crystalline ansolvate, such as a crystalline anhydrous form. The free base of Compound 1, can be obtained from its corresponding sait form, such as the HCl sait of Compound 1.

[0008] Three anhydrous crystalline forms of the free base were identified, termed Free Base Forms I, II, and Material N. It has been discovered that nucléation of Free Base Form I generally occurs first from a slurry. Extending the slurry time can induce the transformation of Free Base Form I to Free Base Form II, a thermodynamically more stable phase relative to Form I. It has further been discovered that Free Base Material N can be stable relative to Forms I and II, at room température.

[0009] Free Base Material N was found to be enantiotropically related to Form II, and will transform reversibly at a spécifie transition température (estimated herein near 40-42 °C). Above the transition température, Free Base Form II appears to be the most stable form, relative to Form I and Material N. Thus, under operating températures below 40 °C, e.g., at 30 °C, the free base of Compound 1 exists primarily as Material N, which may hâve some residual Form IL

Thus, at operating températures above 40 °C, e.g., at 50 °C, the free base of Compound 1 exists primarily as Form II, which may hâve some residual Material N. At 40 °C little appréciable conversion is seen between Material N and Form II. This is contemplated to be true for slurries of the free base in certain solvents and in the solid state. In one embodiment, the one or more crystalline free base forms of Compound 1 do not undergo polymorphie transformation under conditions suitable for manufacturing and storing the crystalline forms.

Form I [0010] In one embodiment, the crystalline free base of Compound 1 comprises crystalline Form I, which is characterized by an endothermie peak at (97±2) C as measured by differential scanning calorimetry. In another embodiment, the crystalline Form I of the free base of crystalline Compound 1 is characterized by the substantial absence of thermal events at températures below the endothermie peak at (97±2) °C as measured by differential scanning calorimetry. In another embodiment, the crystalline Form I of the free base of crystalline Compound 1 is characterized by an X-ray powder diffraction peak (Cu Ka radiation at one or more of 12.82°, 15.74°, 16.03°, 16.63°, 17.60°, 25.14°, 25.82° and 26.44° ±0.2 °2θ. In another embodiment, the crystalline Form I of the free base of crystalline Compound 1 is characterized by an X-ray powder diffraction pattern (Cu Ka radiation) substantially similar to that of FIG. 3.

[0011] In another embodiment, the crystalline Form I of the free base of crystalline Compound 1 is characterized by at least one X-ray powder diffraction peak (Cu Ka radiation) selected from 12.82°, 15.74°, 16.03°, 16.63°, 17.60°, 25.14°, 25.82° and 26.44° (each ±0.2 °29). In another embodiment, the crystalline Form I of the free base of crystalline Compound 1 is characterized by at least two X-ray powder diffraction peaks (Cu Ka radiation) selected from 12.82°, 15.74°, 16.03°, 16.63°, 17.60°, 25.14°, 25.82° and 26.44° (each ±0.2 °2Θ). In another embodiment, the crystalline Form I of the free base of crystalline Compound 1 is characterized by at least three Xray powder diffraction peaks (Cu Ka radiation) selected from 12.82°, 15.74°, 16.03°, 16.63°, 17.60°, 25.14°, 25.82° and 26.44° (each ±0.2 °20).

[0012] In another embodiment, Form I is characterized by 1, 2, 3, 4, or more peaks as tabulated below.

Observed peaks for Form I, XRPD file 609973.

°2θ d space (Â) Intensity (%)

5.52 ± 0.20 16.021 ± 0.602 68

•	12.82 ± 0.20	6.906 ±0.109	74
	15.03 ±0.20	5.897 ± 0.079	38
	15.74 ± 0.20	5.629 ± 0.072	46
	16.03 ± 0.20	5.530 ± 0.069	46
5	16.63 ±0.20	5.331 ±0.064	61
	17.60 ± 0.20	5.040 ± 0.057	100
	18.74 ±0.20	4.736 ± 0.051	24
	19.07 ± 0.20	4.654 ± 0.049	17
	19.35 ± 0.20	4.587 ± 0.047	23
10	20.32 ± 0.20	4.370 ± 0.043	18
	21.64 ±0.20	4.106 ±0.038	23
	22.80 ± 0.20	3.901 ±0.034	26
	23.28 ±0.20	3.821 ±0.033	34
	25.14 ±0.20	3.543 ±0.028	52
15	25.82 ±0.20	3.451 ±0.026	81
	26.44 ± 0.20	3.371 ±0.025	51
	27.91 ±0.20	3.197 ±0.023	17
	28.19 ±0.20	3.165 ±0.022	26

Form II [0013] In another embodiment, the crystalline Compound 1 free base comprises crystalline Form II, which is characterized by an endothermie peak at (97±2) C as measured by differential scanning calorimetry. In another embodiment, the crystalline Form II of the free base of crystalline Compound 1 is characterized by the substantial absence of thermal events at températures below the endothermie peak at (97±2) °C as measured by differential scanning calorimetry. In another embodiment, the crystalline Form II of the free base of crystalline Compound 1 is characterized by an X-ray powder diffraction peak (Cu Ka radiation at one or more of 13.37°, 14.37°, 19.95° or 23.92 °20. In another embodiment, the crystalline Form II of the free base of crystalline Compound 1 is characterized by an X-ray powder diffraction pattern 30 (Cu Ka radiation) substantially similar to that of FIG. 5.

[0014] In another embodiment, the crystalline Form II of the free base of crystalline Compound 1 is characterized by at least one X-ray powder diffraction peak (Cu Ka radiation) selected from 13.37°, 14.37°, 19.95° and23.92°20 (each ±0.2 °20). In another embodiment, the crystalline Form II of the free base of crystalline Compound 1 is characterized by at least two X17658 ray powder diffraction peaks (Cu Ka radiation) selected from 13.37°, 14.37°, 19.95°

and23.92°20 (each ±0.2 °2Θ). In another embodiment, the crystalline Form II of the free base of crystalline Compound 1 is characterized by at least three X-ray powder diffraction peaks (Cu Ka radiation) selected from 13.37°, 14.37°, 19.95° and23.92°20 (each ±0.2 °2Θ).

[0015] In another embodiment, Form II is characterized by 1, 2, 3, 4, or more peaks as tabulated below.

Observed peaks for Form II, XRPD file 613881.

°2θ	d space (Â)	Intensity (%)
10	5.62 ± 0.20	15.735 + 0.581	24
	12.85 ±0.20	6.888 ±0.108	22
	12.97 ± 0.20	6.826 ±0.106	21
	13.37 ±0.20	6.622 ±0.100	100
	14.37 ± 0.20	6.162 ±0.087	56
15	15.31 ±0.20	5.788 ±0.076	21
	16.09 ± 0.20	5.507 ± 0.069	23
	16.45 ± 0.20	5.390 ± 0.066	69
	16.75 ±0.20	5.294 ± 0.064	32
	16.96 ± 0.20	5.227 ± 0.062	53
20	19.95 ±0.20	4.450 ± 0.045	39
	20.22 ± 0.20	4.391 ±0.043	20
	23.18 ±0.20	3.837 ±0.033	38
	23.92 ± 0.20	3.721 ±0.031	41
	24.40 ± 0.20	3.648 ± 0.030	44 .
25	24.73 ± 0.20	3.600 ± 0.029	22
	24.99 ± 0.20	3.564 ± 0.028	50
	25.12 ±0.20	3.545 ±0.028	28
	25.39 ± 0.20	3.509 ±0.027	51
	25.70 ± 0.20	3.466 ±0.027	21
30	26.19 ±0.20	3.403 ±0.026	27
	26.72 ± 0.20	3.336 ±0.025	30
	27.02 ± 0.20	3.300 ±0.024	25
	27.34 ± 0.20	3.262 ±0.024	23
	28.44 ± 0.20	3.138 ±0.022	20

[0016] In some embodiments, the free base of crystalline Compound 1 comprises the crystalline Form II. In some preferred embodiments, the free base of crystalline Compound 1 comprises the crystalline Form II and less than 25 mole %, 10 mole % or 5 mole % of crystalline

Form I, crystalline Material N or amorphous forms of Compound 1.

[0017] In a preferred embodiment, the crystalline Form II is prepared from a slurry comprising the free base of Compound 1 in heptane, from which the crystalline Form II is formed and filtered. Thus, in some embodiments, the crystalline Form II comprises residual (1-500 ppm) heptane. In another preferred embodiment, the crystalline Form II is prepared from a slurry comprising the free base of Compound 1 in water, from which the crystalline Form II is formed and filtered.

[0018] There are several advantages of crystalline Form II relative to crystalline Form I or Material N. For example, the crystalline Form II can be prepared from a slurry comprising the free base of Compound 1 in heptane, which is suitable for good manufacturing practices (GMP) protocols. Further, in a most preferred embodiment, the crystalline Form II can be prepared from a slurry comprising the free base of Compound 1 in water or the HCl sait of Compound 1 in water, thus reducing or eliminating the need for solvent during recrystalization. Thus, in some embodiments, crystalline Form II of Compound 1 comprises less than 500 ppm, 100 ppm, less than 50 ppm or less than 10 ppm organic solvent. Also, Form II has less of a propensity than

Material N to agglomerate upon size réduction, e.g., upon milling. As such, Form II has greater flowability than Material N. Certain illustrative and non-limiting advantages of Form II over Material N (i.e., Form N) are shown in the table below.

DATA/EXPER1MENT	RESULTS/STATUS
Identify suitable solvent for scale-up	Form N: • Limited number of suitable solvents compared to Form II • MTBE identified (suitable for GMP; Class III solvent) • Scale-up results look good Form II: • More solvent options than Form N, including H2O • Current solvent is heptane (suitable for GMP; Class III solvent) • produced on 5 kg scale • Formation time faster than N (could translate to 2-3 day saving in production time) • Better recovery than N
Size/Morphology of N and II	• Acicular morphology observed for form N; material composed of small and large particles • Agglomérâtes are an issue for Form N relative to Form II (less agglomération seen with energy-reduced method)
PK Comparison of N and II	• Oral administrations of GBT440 Forms N and II to rats resulted in comparable exposure at 100 & 500 mg/kg

Material N [0019] In another embodiment, the crystalline Compound 1 free base comprises crystalline

Material N, which is characterized by an endothermie peak at (95±2) C as measured by differential scanning calorimetry. The terms “Material N”, “form N” and “polymorphie form N” are used interchangeably herein. In another embodiment, the crystalline Material N of the free base of crystalline Compound 1 is characterized by the substantial absence of thermal events at températures below the endothermie peak at (95±2) °C as measured by differential scanning calorimetry. In another embodiment, the crystalline Material N of the free base of crystalline Compound 1 is characterized by an X-ray powder diffraction peak (Cu Ka radiation at one or more of 11.65°, 11.85°, 12.08°, 16.70°, 19.65° or 23.48 °20. In another embodiment, the crystalline Material N of the free base of crystalline Compound 1 is characterized by an X-ray powder diffraction pattern (Cu Ka radiation) substantially similar to that of FIG. 7.

[0020] In another embodiment, the crystalline Material N of the free base of crystalline

Compound 1 is characterized by at least one X-ray powder diffraction peak (Cu Ka radiation) selected from 11.65°, 11.85°, 12.08°, 16.70°, 19.65° and 23.48 °20 (each ±0.2 °20). In another embodiment, the crystalline Material N of the free base of crystalline Compound 1 is characterized by at least two X-ray powder diffraction peaks (Cu Ka radiation) selected from 11.65°, 11.85°, 12.08°, 16.70°, 19.65° and 23.48 °2θ (each ±0.2 °20). In another embodiment, the crystalline Material N of the free base of crystalline Compound 1 is characterized by at least three X-ray powder diffraction peaks (Cu Ka radiation) selected from 11.65°, 11.85°, 12.08°, 16.70°, 19.65° and 23.48 °2Θ (each ±0.2 °2Θ).

[0021] In another embodiment, Material N is characterized by 1, 2, 3, 4, or more peaks as tabulated below.

Observed peaks for Material N, XRPD file 615765.

°2θ	d space (Â)	Intensity (%)
5.55 ± 0.20	15.924 ±0.595	54
11.65 ±0.20	7.597 ±0.132	31
11.85 ±0.20	7.468 ±0.128	50
12.08 ± 0.20	7.324 ±0.123	31
12.67 ± 0.20	6.987 ±0.112	29
13.12 ±0.20	6.748 ±0.104	83
14.94 ± 0.20	5.929 ±0.080	34
15.19 ±0.20	5.832 ±0.077	56
15.76 ±0.20	5.623 ± 0.072	20
16.70 ± 0.20	5.310 ±0.064	100
17.35 ±0.20	5.112 ±0.059	52
19.65 ±0.20	4.517 ±0.046	60
23.48 ±0.20	3.789 ±0.032	72
23.68 ±0.20	3.757 ±0.032	29
25.25 ±0.20	3.527 ±0.028	20
25.47 ±0.20	3.497 ±0.027	20
25.70 ± 0.20	3.466 ±0.027	85
26.04 ± 0.20	3.422 ±0.026	35
26.37 ± 0.20	3.380 ± 0.025	55

[0022] In some embodiments, the free base of crystalline Compound 1 comprises the crystalline Material N and less than 25 mole %, 10 mole % or 5 mole % of crystalline Forms I or

II or amorphous forms of Compound 1.

[0023] In another embodiment, the crystalline Material N is prepared from a slurry comprising the free base of Compound 1 in methyl tertiary butyl ether (MTBE), from which the crystalline Material N is formed and filtered. Thus, in some embodiments, the crystalline Material N comprises residual (1-500 ppm) MTBE.

[0024] There are several advantages of crystalline Material N relative to crystalline Forms I or II. For example, the crystalline Material N can be prepared from a slurry comprising the free base of Compound 1 in MTBE, which is suitable for good manufacturing practices (GMP) protocols.

[0025] In some embodiments, the crystalline ansolvate forms are stable to contact with water, heptane, iso propyl ether (IPE), MTBE, and toluene, and such other solvents.

[0026] In another of its composition embodiments, this invention provides for a pharmaceutical composition comprising a pharmaceutically acceptable excipient and a crystalline Compound 1 free base, comprising one or more of Form I, Form II or Material N.

[0027] In one of its method embodiments, this invention provides a method of preparing the solid crystalline free base of Compound 1 comprising, e.g., Form I, Form II and/or Material N.

[0028] In yet another of its method embodiments, there are provided methods for increasing oxygen affinity of hemoglobin S in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of a crystalline free base of Compound 1, comprising, e.g., Form I, Form II and/or Material N.

[0029] In yet another of its method embodiments, there are provided methods for treating oxygen deficiency associated with sickle cell anémia in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of a crystalline free base of Compound 1, comprising, e.g., Form I, Form II and/or Material N.

[0030] In ail of such treatments, the effective amount of free base of Compound 1, comprising e.g., Form I, Form II and/or Material N to the treated patient is already disclosed in the art.

Solvatés [0031] This invention arises in part out of the discovery that ansolvate polymorphs of the free base of Compound 1 form solvaté polymorphs with a variety of solvents, preferably other than certain hydrocarbon solvents, water and ethers.

[0032] Solvatés of the crystalline free base of Compound 1 (e.g., from acetone, acetonitrile, dichloromethane, dioxane, éthanol, ethyl acetate, isopropyl alcohol, methyl ethyl ketone (MEK) and tetrahydrofuran) are also contemplated to be used e.g., as intermediates to regenerate the free base crystalline ansolvate of Compound 1. Such methods can include, without limitation, subjecting the solvaté to vacuum conditions; and/or generating a sait and disproportionating it in water to form the ansolvate ; and/or slurrying or washing the solvaté with a solvent less prone to solvaté formation such as heptane, di-isopropyl ether (IPE), tert-methyl butyl ether (MTBE) and toluene.

[0033] In another of its composition embodiments, this invention provides for a pharmaceutical composition comprising a pharmaceutically acceptable excipient and one or more of the solvated crystal forms provided herein.

[0034] In one of its method embodiments, this invention provides a method of preparing the solvated crystal forms provided herein.

[0035] In yet another of its method embodiments, there are provided methods for increasing oxygen affinity of hemoglobin S in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of one or more of the solvated crystal forms provided herein.

[0036] In yet another of its method embodiments, there are provided methods for treating oxygen deficiency associated with sickle cell anémia in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of one or more of the solvated crystal forms provided herein.

[0037] In ail of such treatments, the effective amount of the free base of Compound 1, to the treated patient is already disclosed in the art.

Brief Description of the Drawings [0038] FIG. 1 is a XRPD profile of the crystalline HCl sait before (top) and after (bottom) 5 minutes slurried in water.

[0039] FIG. 2 is a XRPD profile of the free base Form I (top), Form II (middle), and Material N (bottom).

[0040] FIG. 3 is a XRPD profile and contemplated indexing for free base Form I.

[0041] FIG. 4 is a thermal characterization for free base Form L [0042] FIG. 5 is a XRPD profile and contemplated indexing for free base Form IL [0043] FIG. 6 is a thermal characterization for free base Form II.

[0044]

FIG. 7 is a XRPD profile for free base Material N.

[0045] FIG. 8 is a thermal characterization for free base Material N.

[0046] FIG. 9 depicts an Energy-Temperature Diagram between the Free Base Forms I, II, and Material N. The enthalpy (H) and free energy (G) isobars for each form are depicted as a function of température. ΔΗ/ is the heat of fusion; T is transition température; m is melt température; superscripts I, II, and N refer to the polymorphs. *Under the test conditions, not enough information was available to graphically represent the free energy isobar of Form I below 6 °C and above the estimated transition température T⁷⁷'⁷⁷; the isobar likely intersects G_L at a température below m⁷⁷, allowing the possibility that Form I may be enantiotropic with Form II (where T⁷’⁷⁷ occurs below 6 °C) and/or Material N (where either T¹^ occurs below Z⁷’⁷⁷ or Z⁷⁷⁷ occurs above Z⁷⁷'⁷⁷, but not both). Free energy isobars can only intersect each other once.

[0047] FIG. 10 depicts ¹³C Solid State NMR spectra for Free Base Forms I (bottom), II (middle), and Material N (top). Form I contains one molécule per asymmetric unit. Material N contains four molécules per asymmetric unit. As observed by ¹³C Solid State NMR spectra, Forms II and N did not undergo a transition over 250 K to 340 K. Chemical shifts change slightly with température (not illustrated graphically).

[0048] FIG. 11 depicts ¹⁵N Solid State NMR spectra for Free Base Forms I (bottom), II (middle), and Material N (top).

[0049] FIG. 12 depicts a differential scanning calorimetry (DSC) curve for Free Base Material N.

[0050] FIG. 13 depicts a DSC curve for Free Base Form II.

[0051] FIG. 14 depicts a DSC curve for Free Base Form I.

[0052] FIG. 15 depicts a XRPD profile of maturation experiments for the free base of Compound 1 at multiple températures.

[0053] FIG. 16 depicts a contemplated XRPD profile for solvated Material E.

[0054] FIG. 17 depicts a contemplated XRPD profile for solvated Material F.

[0055] FIG. 18 depicts a contemplated XRPD profile for solvated Material G.

[0056] FIG. 19 depicts a contemplated XRPD profile for solvated Material H.

[0057] FIG. 20 depicts a contemplated XRPD profile for solvated Material J.

[0058] FIG. 21 depicts a contemplated XRPD profile for solvated Material K.

[0059]

FIG. 22 depicts a contemplated XRPD profile for solvated Material L.

[0060] FIG. 23 depicts a contemplated XRPD profile for solvated Material M.

[0061] FIG. 24 depicts a contemplated XRPD profile for solvated Material O.

[0062] FIG. 25 depicts an XRPD profile comparison of contemplated isostructural solvatés of the free base of Compound 1. From top to bottom: Material E from acetone; Material F from ACN; Material G from DCM; Material H from dioxane; Material J from EtOH; Material K from IPA/water (also obtained from IP A); and Material L from THF, Material M from MEK.

Detailed Description [0063] As noted above, this invention is directed, in part, to a stable free base of Compound 1 and, in particular, the free base Form I, Form II or Material N. However, prior to discussing this invention in further detail, the following terms will be defined.

Définitions [0064] As used herein, the following terms hâve the following meanings.

[0065] The singular forms “a,” “an,” and “the” and the like include plural referents unless the context clearly dictâtes otherwise. Thus, for example, reference to “a compound” includes both a single compound and a plurality of different compounds.

[0066] The term “about” when used before a numerical désignation, e.g., température, time, amount, and concentration, including a range, indicates approximations which may vary by ±10 %, ±5 % or ±1 %.

[0067] “Administration” refers to introducing an agent into a patient. A therapeutic amount can be administered, which can be determined by the treating physician or the like. An oral route of administration is preferred. The related terms and phrases administering” and “administration ofwhen used in connection with a compound or pharmaceutical composition (and grammatical équivalents) refer both to direct administration, which may be administration to a patient by a medical professional or by self-administration by the patient, and/or to indirect administration, which may be the act of prescribing a drug. For example, a physician who instructs a patient to self-administer a drug and/or provides a patient with a prescription for a drug is administering the drug to the patient. In any event, administration entails delivery to the patient of the drug.

[0068] The “crystalline ansolvate” of Compound 1 is a crystalline solid form of the free base of 2-hydroxy-6-((2-( 1 -isopropyl-lH-pyrazol-5-yl)pyridin-3-yl)methoxy)benzaldehyde, such as,

e.g,, crystalline Form I, Form II or Material N as disclosed herein. Each of the Form I, Form II or

Material N crystal lattices is substantially free of solvents of crystallization. However, any solvent présent is not included in the crystal lattice and is randomly distributed outside the crystal lattice. Therefore, Form I, Form II or Material N crystals in bulk may contain, outside the 5 crystal lattice, small amounts of one or more solvents, such as the solvents used in its synthesis or crystallization. As used above, “substantially free of ’ and “small amounts,” refers to the presence of solvents preferably less that 10,000 parts per million (ppm), or more preferably, less than 500 ppm.

[0069] The “crystalline solvaté” of Compound 1 is a crystalline solid form of the free base of 10 2-hydroxy-6-((2-(l-isopropyl-lH-pyrazol-5-yl)pyridin-3-yl)methoxy)benzaldehyde, where the crystal lattices comprises one or more solvents of crystallization.

[0070] “Characterization” refers to obtaining data which may be used to identify a solid form of a compound, for example, to identify whether the solid form is amorphous or crystalline and whether it is unsolvated or solvated. The process by which solid forms are characterized involves 15 analyzing data collected on the polymorphie forms so as to allow one of ordinary skill in the art to distinguish one solid form from other solid forms containing the same material. Chemical identity of solid forms can often be determined with solution-state techniques such as ¹³C NMR or 'H NMR. While these may help identify a material, and a solvent molécule for a solvaté, such solution-state techniques themselves may not provide information about the solid state. There 20 are, however, solid-state analytical techniques that can be used to provide information about solid-state structure and differentiate among polymorphie solid forms, such as single crystal X-ray diffraction, X-ray powder diffraction (XRPD), solid state nuclear magnetic résonance (SS-NMR), and infrared and Raman spectroscopy, and thermal techniques such as differential scanning calorimetry (DSC), Solid state ¹³C-NMR, thermogravimetry (TG), melting point, and 25 hot stage microscopy.

[0071] To “characterize” a solid form of a compound, one may, for example, collect XRPD data on solid forms of the compound and compare the XRPD peaks of the forms. For example, when only three solid forms, e.g., Forms I and II and Material N, are compared and the Form I pattern shows a peak at an angle where no peaks appear in the Form II or Material N pattern, then that peak, for that compound, distinguishes Form I from Form II and Material N and further acts to characterize Form I. The collection of peaks which distinguish e.g., Form I from the other known forms is a collection of peaks which may be used to characterize Form I. Those of ordinary skill in the art will recognize that there are often multiple ways, including multiple ways using the same analytical technique, to characterize solid forms. Additional peaks could also be

used, but are not necessary, to characterize the form up to and including an entire diffraction pattern. Although ail the peaks within an entire XRPD pattern may be used to characterize such a form, a subset of that data may, and typically is, used to characterize the form.

[0072] An XRPD pattern is an x-y graph with diffraction angle (typically ° 2Θ) on the x-axis and intensity on the y-axis. The peaks within this pattern may be used to characterize a crystalline solid form. As with any data measurement, there is variability in XRPD data. The data are often represented solely by the diffraction angle of the peaks rather than including the intensity of the peaks because peak intensity can be particularly sensitive to sample préparation 10 (for example, particle size, moisture content, solvent content, and preferred orientation effects influence the sensitivity), so samples of the same material prepared under different conditions may yield slightly different patterns; this variability is usually greater than the variability in diffraction angles. Diffraction angle variability may also be sensitive to sample préparation. Other sources of variability corne from instrument parameters and processing of the raw X-ray 15 data: different X-ray instruments operate using different parameters and these may lead to slightly different XRPD patterns from the same solid form, and similarly different software packages process X-ray data differently and this also leads to variability. These and other sources of variability are known to those of ordinary skill in the pharmaceutical arts. Due to such sources of variability, it is usual to assign a variability of ±0.2° 2Θ to diffraction angles in XRPD patterns.

[0073] “Comprising” or “comprises” is intended to mean that the compositions and methods include the recited éléments, but not exclude others. “Consisting essentially of ’ when used to define compositions and methods, shall mean excluding other éléments of any essential significance to the combination for the stated purpose. Thus, a composition consisting essentially 25 of the éléments as defined herein would not exclude other materials or steps that do not materially affect the basic and novel characteristic(s) of the claimed invention. “Consisting of ’ shall mean excluding more than trace éléments of other ingrédients and substantial method steps. Embodiments defined by each of these transition terms are within the scope of this invention.

[0074] Form II and Material N are enantiotropic at a transition température (of approximately 30 42 °C). Below this transition température, Material N of the free base of Compound 1 is the thermodynamically more stable form relative to Forms I and II. Above this transition température, Form II of the free base of Compound 1 is the thermodynamically more stable form relative to Form I and Material N.

[0075] “Room température” refers to (22±5) °C.

[0076] “Therapeutically effective amount” or “therapeutic amount” refers to an amount of a drug or an agent that when administered to a patient suffering from a condition, will hâve the intended therapeutic effect, e.g., alleviation, amelioration, palliation or élimination of one or more manifestations of the condition in the patient. The therapeutically effective amount will vary depending upon the subject and the condition being treated, the weight and âge of the subject, the severity of the condition, the particular composition or excipient chosen, the dosing regimen to be followed, timing of administration, the manner of administration and the like, ail of which can be determined readily by one of ordinary skill in the art. The full therapeutic effect does not necessarily occur by administration of one dose, and may occur only after administration of a sériés of doses. Thus, a therapeutically effective amount may be administered in one or more administrations. For example, and without limitation, a therapeutically effective amount of an agent, in the context of treating disorders related to hemoglobin S, refers to an amount of the agent that alleviates, améliorâtes, palliâtes, or éliminâtes one or more manifestations of the disorders related to hemoglobin S in the patient.

[0077] “Treatment”, “treating”, and “treat” are defined as acting upon a disease, disorder, or condition with an agent to reduce or ameliorate the harmful or any other undesired effects of the disease, disorder, or condition and/or its symptoms. Treatment, as used herein, covers the treatment of a human patient, and includes: (a) reducing the risk of occurrence of the condition in a patient determined to be predisposed to the disease but not yet diagnosed as having the condition, (b) impeding the development of the condition, and/or (c) relieving the condition, i.e., causing régression of the condition and/or relieving one or more symptoms of the condition. For purposes of this invention, bénéficiai or desired clinical results include, but are not limited to, multilineage hématologie improvement, decrease in the number of required blood transfusions, decrease in infections, decreased bleeding, and the like.

Identifying Forms I, II and Material N [0078] When the HCl sait of Compound 1 was subjected to various stress conditions, disproportionation of the HCl sait in water was observed to generate the free base. At least three anhydrous crystalline forms of the free base were identified, termed Free Base Forms I, II, and

Material N. It was discovered that nucléation of Free Base Form I generally occurs first and that extending the slurry time induces the transformation of Free Base Form I to Free Base Form II, a more thermodynamically stable phase relative to Form I. It was further discovered that Free

Base Material N appears to be most stable form, relative to Forms I and II, at room température.

Free Base Material N was found to be enantiotropically active relative to Form II, and will transform reversibly at a spécifie transition température (estimated herein near 42 °C). Above the transition température, Free Base Form II appears to be the most stable form, relative to Form I and Material N.

[0079] Based in part on solid-state nuclear magnetic résonance data, ail three forms are crystalline and are distinct polymorphie forms. See FIGS 10 and 11. Form I contains one molécule per asymmetric unit, Form II contains two molécules per asymmetric unit and Form N contains four molécules per asymmetric unit. See the ¹⁵N spectra in FIG. 11.

Ansolvates of Forms I, II and Material N [0080] In one embodiment, this invention provides the free base crystalline ansolvate of Compound 1. The free base crystalline ansolvate of Compound 1 may include one or more of Form I, Form II and/or Material N polymorphs. In some embodiments, the free base crystalline ansolvate of Compound 1 may include the Form II polymorph. Preferably, the free base crystalline ansolvate of Compound 1 may include Form II and/or Material N polymorphs. More preferably, the free base crystalline ansolvate of Compound 1 may include the Material N polymorph. Yet more preferably, the free base crystalline ansolvate of Compound 1 is substantially free of a solvated polymorph of Compound 1 free base. Further yet more preferably, the free base crystalline ansolvate of Compound 1 is substantially free of other ansolavte polymorphs of Compound 1 free base. “Substantially free” of a component as used herein refers to contain up to about 5%, more preferably about 3%, and still more preferably about 1% of that coponent. As used herein, solvaté includes a hydrate form as well.

Solvatés of Compound 1 [0081] In one aspect, provided is a crystalline solvaté of Compound 1:

ΌΗ.

Compound 1 [0082] In some embodiments, the crystalline solvaté is substantially free of an ansolvated polymorph of Compound 1.

[0083] Many of the solubility and screen experiments with the free base of Compound 1 resulted in précipitation of solids characterized as solvaté formation with some solvents. Under the conditions, solvatés were not observed from the free base of Compound 1 with four solvents, including heptane, di-isopropyl ether (IPE), tert-methyl butyl ether (MTBE) and toluene. Solvatés were observed from the free base of Compound 1 in nine solvents including acetone (Material E), acetonitrile (Material F), dichloromethane (Material G), dioxane (Material H), éthanol (Material J), isopropyl alcohol or a mixture of water and isopropyl alcohol (Material K), tetrahydrofuran (Material L), methyl ethyl ketone “MEK” (Material M), ethyl acetate (Material O) and dimethyl sulfoxide “DMSO” (Material P). The majority of the solvatés (i.e., Materials EH, J-M, O and P are contemplated to be isostructural. In some embodiments, the crystalline solvaté includes one or more of Material E, Material F, Material G, Material H, Material J, Material K, Material L, Material M, Material O or Material P.

[0084] Material E can be characterized by at least one X-ray powder diffraction peak (Cu Ka radiation) selected from 8.69, 11.73, 12.10, 15.26, 16.11, 17.45, 22.39, 22.55 and 23.70 ± 0.20. Material F can be characterized by at least one X-ray powder diffraction peak (Cu Ka radiation) selected from 8.47, 8.81, 12.75, 13.17, 14.92, 15.63, 17.01 23.73, and 24.07 ± 0.20. Material G can be characterized by at least one X-ray powder diffraction peak (Cu Ka radiation) selected from 8.47, 11.45, 12.62, 14.66, 15.69, 17.01, 18.47, 20.32, 22.61, 23.08, 23.43 and 23.70 ± 0.20. Material H can be characterized by at least one X-ray powder diffraction peak (Cu Ka radiation) selected from 8.61, 11.67, 15.33, 16.28, 17.28,22.58,23.51 and 25.77 ± 0.20. Material J can be characterized by at least one X-ray powder diffraction peak (Cu Ka radiation) selected from 8.52,8.88, 12.79, 15.04, 15.61, 17.11, 22.81, 23.87, 24.17, 24.62 and 26.44 ± 0.20. Material K can be characterized by at least one X-ray powder diffraction peak (Cu Ka radiation) selected from 8.52; 8.83,11.35, 15.04,15.74,17.11, 23.46, 23.58, 24.08 and 25.99 ± 0.20. Material L can be characterized by at least one X-ray powder diffraction peak (Cu Ka radiation) selected from 8.61, 8.78, 11.67, 14.94,15.28,16.14,17.30, 22.75, 23.71 and 26.05 ± 0.20; and Material M can be characterized by at least one X-ray powder diffraction peak (Cu Ka radiation) selected from 7.74, 10.05, 12.82, 15.33, 16.80, 20.82, 21.14, 25.80 and 26.97 ± 0.20.

[0085] The solvatés (such as, of acetone, acetonitrile, dichloromethane, dioxane, éthanol, ethyl acetate, isopropyl alcohol, MEK, tetrahydrofuran or DMSO) could be used e.g., as intermediates to regenerate the free base crystalline ansolvate of Compound 1 by several methods including subjecting the solvaté to vacuum conditions; and/or regenerating the HCl sait and dispiOportionating HCl; and/or washing the solvaté with a solvent less prone to solvaté formation such as heptane, di-isopropyl ether (IPE), tert-methyl butyl ether (MTBE) and toluene.

Table 1. Data Related to Solvatés of the Free Base of Compound 1

Identifier	Crystallization Solvent	Volume (Â3/Cell)	Number of Formula Units per Cell	Estimated Volume per Formula Unit* (A3)	Indexing Resuit
Material E	acetone	968	2	484	FIG 1
Material F	ACN	947	2	473	FIG 2
Material G	DCM	959	2	480	FIG 3
Material H	dioxane	977	2	488	FIG 4
Material J	EtOH	943	2	472	FIG 5
Material K	IPA	963	2	481	FIG 6
Material L	THF	972	2	486	FIG 7
Material M	MEK	3956	8	494	FIG 8
Material O	EtOAc	-	-	-	FIG 9
Material p**	DMSO			-	-

* The value is obtained by dividing the volume of the cell, derived from the tentative indexing solution, by the number of formula units within the cell. ** Material P was observed as a mixture with a “sulfate form I”.

[0086] Certain contemplated peaks of the various solvatés provided herein are tabulated below. Certain peaks, which are preferably non-overlapping, low-angle peaks, with strong intensity, were not identified. The peaks were determined to the extent that the state of preferred orientation in the samples were unknown.

Table 2. Observed peaks for Material E.

°2Θ	d space (Â)	Intensity (%)
8.41 ± 0.20	10.517 ±0.256	13
8.69 ±0.20	10.174 ±0.239	100
11.73 ±0.20	7.543 ±0.130	17
12.10 ±0.20	7.314 ±0.122	20
13.00 ±0.20	6.809 ±0.106	15
14.02 ± 0.20	6.316 ±0.091	5
14.77 ± 0.20	5.996 ± 0.082	16
15.26 ±0.20	5.807 ± 0.077	34
15.81 ±0.20	5.605 ±0.071	7

•	16.11 ±0.20	5.501 ± 0.069	20
	16.48 ± 0.20	5.379 ± 0.066	11
	16.65 ± 0.20	5.326 ± 0.064	11
	16.88 ±0.20	5.253 ±0.063	3
5	17.26 ± 0.20	5.136 ±0.060	9
	17.45 ± 0.20	5.083 ± 0.058	32
	20.02 ± 0.20	4.435 ± 0.044	2
	20.92 ± 0.20	4.246 ± 0.041	13
	21.91 ±0.20	4.057 ± 0.037	20
10	22.39 ± 0.20	3.970 ±0.035	49
	22.55 ± 0.20	3.944 ± 0.035	37
	22.81 ±0.20	3.898 ±0.034	16
	23.36 ±0.20	3.807 ±0.032	12
	23.70 ±0.20	3.755 ±0.032	61
15	24.37 ± 0.20	3.653 ± 0.030	12
	24.85 ± 0.20	3.583 ±0.029	5
	25.42 ± 0.20	3.504 ± 0.027	2
	25.89 ± 0.20	3.442 ±0.026	8
	26.19 ±0.20	3.403 ±0.026	40
20	26.97 ± 0.20	3.306 ±0.024	3
	27.61 ± 0.20	3.231 ±0.023	16
	28.24 ± 0.20	3.160 ±0.022	2
	28.48 ± 0.20	3.134 ±0.022	5
	28.69 ± 0.20	3.111 ±0.021	7
25	29.83 ± 0.20	2.995 ± 0.020	4

Table 3. Observed peaks for Material F.

°20	d space (À)	Intensity (%)
8.47 ± 0.20	10.434 ± 0.252	100
8.81 ±0.20	10.039 ± 0.233	49
11.42 ±0.20	7.752 ±0.138	15
12.75 ± 0.20	6.942 ±0.110	27
13.17 ±0.20	6.723 ±0.103	21
13.87 ±0.20	6.384 ±0.093	7

•	14.61 ±0.20	6.064 ± 0.084	13
	14.92 ±0.20	5.936 ± 0.080	43
	15.51 ±0.20	5.713 ±0.074	24
	15.63 ± 0.20	5.671 ±0.073	43
5	15.96 ±0.20	5.553 ± 0.070	15
	17.01 ±0.20	5.212 ± 0.062	31
	17.26 ±0.20	5.136 ±0.060	4
	17.70 ±0.20	5.011 ±0.057	9
	18.17 ±0.20	4.883 ± 0.054	4
10	18.79 ±0.20	4.724 ± 0.050	10
	19.35 ±0.20	4.587 ± 0.047	4
	19.49 ± 0.20	4.555 ± 0.047	3
	20.02 ± 0.20	4.435 ± 0.044	4
	20.29 ± 0.20	4.377 ± 0.043	9
15	21.06 ±0.20	4.219 ± 0.040	11
	21.33 ±0.20	4.167 ±0.039	4
	22.71 ±0.20	3.915 ±0.034	27
	23.11 ±0.20	3.848 ±0.033	15
	23.73 ± 0.20	3.749 ±0.031	42
20	24.07 ± 0.20	3.698 ±0.031	59
	24.65 ± 0.20	3.612 ±0.029	87
	24.95 ± 0.20	3.569 ±0.028	6
	25.20 ± 0.20	3.534 ±0.028	5
	25.69 ± 0.20	3.468 ± 0.027	15
25	26.52 ± 0.20	3.361 ±0.025	61
	26.79 ±0.20	3.328 ±0.025	10
	27.02 ± 0.20	3.300 ±0.024	9

Table 4. Observed peaks for Material G.

30	°2Θ	d space (Â)	Intensity (%)
	8.47 ±0.20	10.434 ± 0.252	45
	8.76 ± 0.20	10.096 ± 0.235	12
	11.45 ±0.20	7.729 ±0.137	76
	12.62 ±0.20	7.015 ±0.113	36
35	13.09 ±0.20	6.765 ±0.105	10

13.87 ±0.20	6.384 ±0.093	5
14.66 ± 0.20	6.044 ± 0.083	39
14.92 ± 0.20	5.936 ± 0.080	26
15.33 ±0.20	5.782 ± 0.076	7
15.69 ±0.20	5.647 ± 0.072	88
16.01 ±0.20	5.536 ± 0.070	8
16.76 ± 0.20	5.289 ± 0.063	15
17.01 ±0.20	5.212 ±0.062	29
17.50 ±0.20	5.068 ± 0.058	5
17.60 ± 0.20	5.040 ± 0.057	4
18.13 ±0.20	4.892 ± 0.054	5
18.47 ±0.20	4.804 ± 0.052	21
19.55 ± 0.20	4.540 ± 0.046	4
20.01 ±0.20	4.439 ± 0.044	5
20.32 ± 0.20	4.370 ± 0.043	20
21.11 ±0.20	4.209 ± 0.040	15
22.61 ± 0.20	3.932 + 0.035	42
22.88 ±0.20	3.887 ±0.034	9
23.08 ±0.20	3.854 ±0.033	28
23.43 ± 0.20	3.797 ± 0.032	56
23.70 ± 0.20	3.755 ±0.032	48
24.12 ±0.20	3.690 ± 0.030	13
24.42 ± 0.20	3.646 ± 0.030	100
25.05 ± 0.20	3.555 ±0.028	7
25.40 ± 0.20	3.506 ± 0.027	26
26.36 ± 0.20	3.382 ±0.025	50
26.57 ± 0.20	3.355 ±0.025	7
26.82 ± 0.20	3.324 ±0.025	27
27.07 ± 0.20	3.294 ±0.024	10

Table 5. Observed peaks for Material H.
°2Θ	dspace (Â)	Intensity (%)
8.61 ±0.20	10.273 ± 0.244	48
8.81 ±0.20	10.039 ± 0.233	20
11.67 ±0.20	7.586 ±0.132	32

12.79 ± 0.20	6.924 ±0.110	9
14.56 ± 0.20	6.085 ± 0.084	4
14.87 ± 0.20	5.956 ±0.081	22
15.33 ±0.20	5.782 ± 0.076	42
15.76 ±0.20	5.623 ± 0.072	18
16.28 ±0.20	5.445 ± 0.067	51
16.73 ± 0.20	5.299 ± 0.064	9
17.28 ± 0.20	5.132 ±0.060	61
17.68 ± 0.20	5.016 ± 0.057	3
20.47 ± 0.20	4.338 ± 0.042	12
21.38 ±0.20	4.157 ±0.039	7
21.83 ±0.20	4.072 ± 0.037	4
22.23 ± 0.20	3.999 ±0.036	9
22.58 ± 0.20	3.938 ±0.035	100
22.95 ± 0.20	3.876 ±0.034	6
23.11 ±0.20	3.848 ±0.033	14
23.51 ±0.20	3.783 ±0.032	88
24.37 ± 0.20	3.653 ±0.030	13
24.65 ± 0.20	3.612 ±0.029	10
25.77 ± 0.20	3.457 ±0.027	41
26.67 ± 0.20	3.342 ±0.025	7
26.97 ± 0.20	3.306 ± 0.024	5
27.66 ± 0.20	3.225 ±0.023	3
28.11 ±0.20	3.174 ±0.022	4
28.61 ± 0.20	3.120 ±0.022	6
28.96 ± 0.20	3.083 ±0.021	4
29.23 ± 0.20	3.055 ±0.021	3
29.63 ± 0.20	3.015 ±0.020	3

Table 6. Observed peaks for Material J.
°2θ	d space (Â)	Intensity (%)
8.52 ± 0.20	10.373 ± 0.249	100
8.88 ±0.20	9.964 ± 0.229	39
11.33 ±0.20	7.809 ±0.140	22

•	12.79 ± 0.20	6.924 ±0.110	25
	13.12 ±0.20	6.748 ±0.104	24
	13.94 ±0.20	6.354 ± 0.092	4
	14.47 ± 0.20	6.120 ±0.085	14
5	15.04 ±0.20	5.890 ± 0.079	42
	15.61 ±0.20	5.677 ±0.073	56
	15.84 ±0.20	5.594 ±0.071	16
	17.11 ±0.20	5.181 ±0.061	33
	17.40 ± 0.20	5.097 ± 0.059	4
10	17.82 ±0.20	4.979 ± 0.056	8
	18.12 ±0.20	4.897 ±0.054	3
	18.90 ±0.20	4.695 ± 0.050	11
	19.39 ±0.20	4.579 ± 0.047	5
	19.62 ± 0.20	4.525 ± 0.046	4
15	20.16 ±0.20	4.406 ± 0.044	8
	20.96 ± 0.20	4.239 ± 0.040	12
	22.81 ±0.20	3.898 ±0.034	27
	23.15 ±0.20	3.843 ±0.033	9
	23.28 ±0.20	3.821 ±0.033	7
20	23.87 ±0.20	3.729 ±0.031	34
	24.17 ±0.20	3.683 ±0.030	52
	24.62 ± 0.20	3.616 ±0.029	95
	25.20 ± 0.20	3.534 ±0.028	5
	25.77 ± 0.20	3.457 ±0.027	13
25	26.44 ± 0.20	3.371 ±0.025	70
	26.71 ±0.20	3.338 ±0.025	10
	27.21 ± 0.20	3.278 ±0.024	7

Table 7. Observed peaks for GBT000440, Material K.

30	°20	dspace(Â)	Intensity (%)
	8.52 ± 0.20	10.373 ± 0.249	75
	8.83 ± 0.20	10.020 ± 0.232	33
	11.35 ±0.20	7.797 ±0.139	29
	12.52 ± 0.20	7.071 ±0.114	21
35	12.90 ± 0.20	6.861 ±0.108	24

•
	13.92 ±0.20	6.361 ±0.092	4
	14.49 ± 0.20	6.113 ±0.085	18
	15.04 ±0.20	5.890 ± 0.079	41
	15.34 ±0.20	5.775 ± 0.076	17
5	15.74 ± 0.20	5.629 ± 0.072	57
	15.93 ±0.20	5.564 ± 0.070	13
	16.61 ±0.20	5.336 ± 0.065	7
	17.11 ±0.20	5.181 ±0.061	33
	17.70 ± 0.20	5.011 ±0.057	7
10	18.00 ±0.20	4.928 ± 0.055	4
	18.38 ±0.20	4.826 ± 0.053	13
	19.04 ± 0.20	4.662 ± 0.049	4
	19.74 ± 0.20	4.498 ± 0.046	5
	20.21 ±0.20	4.395 ± 0.043	11
15	20.99 ± 0.20	4.232 ± 0.040	12
	22.70 ± 0.20	3.918 ±0.034	22
	22.90 ±0.20	3.884 ±0.034	17
	23.46 ± 0.20	3.791 ±0.032	45
	23.58 ± 0.20	3.773 ±0.032	70
20	24.08 ± 0.20	3.695 ±0.030	100
	24.75 ± 0.20	3.597 ±0.029	6
	25.19 ±0.20	3.536 ±0.028	21
	25.99 ± 0.20	3.429 ±0.026	71
	26.71 ±0.20	3.338 ±0.025	11
25	27.36 ± 0.20	3.260 ±0.024	9
	28.11 ±0.20	3.174 ±0.022	4
	28.69 ± 0.20	3.111 ±0.021	9

Table 8. Observed peaks for Material L.

30	°2Θ	d space (Â)	Intensity (%)
	8.61 ±0.20	10.273 ± 0.244	79
	8.78 ± 0.20	10.077 ± 0.235	38
	11.67 ±0.20	7.586 ±0.132	35
	12.17 ±0.20	7.274 ±0.121	19
35	12.94 ± 0.20	6.844 ±0.107	14

•	14.07 ± 0.20	6.293 ± 0.090	3
	14.62 ± 0.20	6.057 ± 0.084	5
	14.94 ± 0.20	5.929 ± 0.080	25
	15.28 ± 0.20	5.800 ± 0.076	50
5	15.93 ± 0.20	5.564 ± 0.070	18
	16.14 ±0.20	5.490 ± 0.068	49
	16.33 ±0.20	5.429 ± 0.067	9
	16.70 ±0.20	5.310 ±0.064	9
	16.85 ± 0.20	5.263 ± 0.063	6
10	17.30 ± 0.20	5.127 ±0.060	52
	17.63 ±0.20	5.030 ±0.057	6
	18.37 + 0.20	4.830 ± 0.053	3
	20.14 ±0.20	4.409 ± 0.044	5
	20.59 ± 0.20	4.314 ±0.042	14
15	21.53 ±0.20	4.128 ±0.038	11
	22.01 ± 0.20	4.038 ±0.037	3
	22.44 ± 0.20	3.961 ±0.035	27
	22.75 ± 0.20	3.910 ±0.034	72
	23.10 ±0.20	3.851 ±0.033	20
20	23.31 ±0.20	3.816 ±0.033	19
	23.48 ± 0.20	3.789 ±0.032	12
	23.71 ±0.20	3.752 ±0.031	100
	24.48 ± 0.20	3.636 ±0.029	20
	24.70 ± 0.20	3.604 ±0.029	4
25	24.93 ± 0.20	3.571 ±0.028	3
	25.59 ± 0.20	3.482 ±0.027	5
	25.72 ±0.20	3.464 ±0.027	5
	26.05 ± 0.20	3.420 ±0.026	62
	26.59 ± 0.20	3.352 ±0.025	6
30	27.14 ±0.20	3.286 ±0.024	8
	27.83 ± 0.20	3.206 ±0.023	8
	28.38 ±0.20	3.145 ±0.022	3
	28.78 ± 0.20	3.102 ±0.021	8
	29.05 ± 0.20	3.074 ± 0.021	4
35	29.36 ± 0.20	3.042 ±0.020	3

Table 9. Observed peaks for Material M.

°2Θ 7.74 ± 0.20	o dspace (A) 11.424 ±0.303	Intensity (%) 100
5	8.34 ± 0.20	10.601 ± 0.260	4
	10.05 ± 0.20	8.806 ±0.178	17
	12.82 ± 0.20	6.906 ±0.109	46
	13.05 ±0.20	6.783 ±0.105	4
	14.17 ±0.20	6.249 ± 0.089	2
10	14.54 ± 0.20	6.092 ± 0.085	6
	14.99 ± 0.20	5.910 ± 0.079	16
	15.33+0.20	5.782 ± 0.076	47
	15.53 ±0.20	5.707 ±0.074	21
	16.80 ± 0.20	5.278 ± 0.063	27
15	18.33 ±0.20	4.839 ±0.053	3
	19.17 ±0.20	4.630 ± 0.048	22
	20.19 ±0.20	4.399 ± 0.044	23
	20.82 ± 0.20	4.266 ± 0.041	32
	21.14 ±0.20	4.202 ± 0.040	27
20	21.29 ±0.20	4.173 ±0.039	14
	22.01 ± 0.20	4.038 ±0.037	13
	22.28 ± 0.20	3.991 ±0.036	23
	22.93 ± 0.20	3.879 ±0.034	6
	23.35 ± 0.20	3.810 ±0.032	11
25	24.00 ± 0.20	3.708 ±0.031	14
	24.25 ± 0.20	3.670 ±0.030	3
	24.88 ± 0.20	3.578 ±0.029	11
	25.54 ± 0.20	3.488 ±0.027	9
	25.80 ± 0.20	3.453 ± 0.027	94
30	26.97 ± 0.20	3.306 ± 0.024	27
	27.63 ± 0.20	3.229 ±0.023	2
	28.41 ±0.20	3.142 ±0.022	7
	28.54 ± 0.20	3.127 ±0.022	8
	29.03 ± 0.20	3.076 ±0.021	3
35	29.30 ± 0.20	3.049 ±0.020	7

29.63 ± 0.20

3.015 ±0.020 15

Pharmaceutical Compositions [0087] In another of its composition embodiments, this invention provides for a pharmaceutical composition comprising a pharmaceutically acceptable excipient and crystalline free base ansolvate of Compound 1, preferably including one or more of the Form I, Form II and/or Material N polymorphs.

[0088] Such compositions can be formulated for different routes of administration. Although compositions suitable for oral delivery will probably be used most frequently, other routes that may be used include intravenous, intraarterial, pulmonary, rectal, nasal, vaginal, lingual, intramuscular, intraperitoneal, intracutaneous, intracranial, subcutaneous and transdermal routes. Suitable dosage forms for administering any of the compounds described herein include tablets, capsules, pills, powders, aérosols, suppositories, parenterals, and oral liquids, including suspensions, solutions and émulsions. Sustained release dosage forms may also be used, for example, in a transdermal patch form. Ail dosage forms may be prepared using methods that are standard in the art (see e.g., Remington’s Pharmaceutical Sciences, 16^th ed., A. Oslo editor, EastonPa. 1980).

[0089] Pharmaceutically acceptable excipients are non-toxic, aid administration, and do not adversely affect the therapeutic benefit of the compound of this invention. Such excipients may be any solid, liquid, semi-solid or, in the case of an aérosol composition, gaseous excipient that 20 is generally available to one of skill in the art. Pharmaceutical compositions in accordance with the invention are prepared by conventional means using methods known in the art.

[0090] The compositions disclosed herein may be used in conjunction with any of the vehicles and excipients commonly employed in pharmaceutical préparations, e.g., talc, gum arabic, lactose, starch, magnésium stéarate, cocoa butter, aqueous or non-aqueous solvents, oils, paraffin 25 dérivatives, glycols, etc. Coloring and flavoring agents may also be added to préparations, particularly to those for oral administration. Solutions can be prepared using water or physiologically compatible organic solvents such as éthanol, 1,2-propylene glycol, polyglycols, dimethylsulfoxide, fatty alcohols, triglycérides, partial esters of glycerin and the like.

[0091] Solid pharmaceutical excipients include starch, cellulose, hydroxypropyl cellulose, talc, 30 glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, magnésium stéarate, sodium stéarate, glycerol monostearate, sodium chloride, dried skim milk and the like. Liquid and semisolid excipients may be selected from glycerol, propylene glycol, water, éthanol and various oils, including those of petroleum, animal, vegetable or synthetic origin, e.g., peanut oil, soybean

oil, minerai oil, sesame oil, etc. In certain embodiments, the compositions provided herein comprises one or more of α-tocopherol, gum arabic, and/or hydroxypropyl cellulose.

[0092] In one embodiment, this invention provides sustained release formulations such as drug depots or patches comprising an effective amount of a compound provided herein. In another embodiment, the patch further comprises gum Arabic or hydroxypropyl cellulose separately or in combination, in the presence of alpha-tocopherol. Preferably, the hydroxypropyl cellulose has an average MW of from 10,000 to 100,000. In a more preferred embodiment, the hydroxypropyl cellulose has an average MW of from 5,000 to 50,000.

[0093] Compounds and pharmaceutical compositions of this invention maybe used alone or in combination with other compounds. When administered with another agent, the coadministration can be in any manner in which the pharmacological effects of both are manifest in the patient at the same time. Thus, co-administration does not require that a single pharmaceutical composition, the same dosage form, or even the same route of administration be used for administration of both the compound of this invention and the other agent or that the two agents be administered at precisely the same time. However, co-administration will be accomplished most conveniently by the same dosage form and the same route of administration, at substantially the same time. Obviously, such administration most advantageously proceeds by delivering both active ingrédients simultaneously in a novel pharmaceutical composition in accordance with the présent invention.

Préparative and Treatment Methods

Ansolvate [0094] In another aspect, the présent invention provides a method of preparing the crystalline free base ansolvate of Compound 1. In one embodiment, provided herein is a method of preparing the crystalline free base of Compound 1 comprising slurrying or contacting the HCl sait of the Compound 1 with water and allowing dissociation of HCl to produce the free base of Compound 1. In one embodiment, the crystalline free base ansolavte of Compound 1 prepared comprises one or more of Form I, Form II and Material N.

[0095] In yet another of its method embodiments, there are provided methods for increasing oxygen affinity of hemoglobin S in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of a crystalline free base of Compound 1. In some embodiments, the crystalline free base of Compound 1 is an ansolvate. In one embodiment, the crystalline free base of Compound 1 comprises one or more of Form I, Form II and Material N.

[0096] In yet another of its method embodiments, there are provided methods for treating oxygen deficiency associated with sickle cell anémia in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of a crystalline free base of Compound 1. In some embodiments, the crystalline free base of Compound 1 is an ansolvate. In one embodiment, the crystalline free base of Compound 1 comprises one or more of Form I, Form II and Material N.

[0097] In further aspects of the invention, a method is provided for treating sickle cell disease, the method comprising administering to a subject in need thereof a therapeutically effective amount of a crystalline free base of Compound 1. In some embodiments, the crystalline free base of Compound 1 is an ansolvate. In one embodiment, the crystalline free base of Compound 1 comprises one or more of Form I, Form II and Material N. In still further aspects of the invention, a method is provided for treating cancer, a pulmonary disorder, stroke, high altitude sickness, an ulcer, a pressure sore, Alzheimer’s disease, acute respiratory disease syndrome, and a wound, the method comprising administering to a subject in need thereof a therapeutically effective amount of a crystalline free base of Compound 1. In some embodiments, the crystalline free base of Compound 1 is an ansolvate. In one embodiment, the crystalline free base of Compound 1 comprises one or more of Form I, Form II and Material N.

[0098] In such treatments, the dosing of the crystalline free base of Compound 1 to the treated patient is already disclosed in the art.

Solvatés [0099] In another aspect, the présent invention provides a method of preparing the crystalline free base solvatés of Compound 1. In some embodiments, a free base ansolvate, as described herein (e.g, as obtained by slurrying an HCl sait of Compound 1 in water) of Compound 1 is contacted with a solvent as provided herein, including a mixture of solvents, to préparé the solvaté, the solvent or the mixture of solvents. Thus, a solvent can be a single solvent or substantially a single solvent or a mixture of solvents. When a mixture of solvents is used, a solvaté can be produced having one or more of the individual constituent solvents of the solvent mixture. In some embodiments, the solvent includes alcoholic solvents such as mono di or higher alcohols or alkanols. In some embodiments, the solvent includes chlorinated solvents such as dichloromethane chloroform, et cetera. In some embodiments, the solvent includes ketone solvents such as alkanones and cycloalkanones. Certain solvents include without limitation, methanol, éthanol, 2-propanol, 2-methyl-1-propanol, 1-butanol, acetonitrile, acetone, dichloromethane, dioxane, or tetrahydrofuran, or combinations thereof, optionally including water.

[0100] In another aspect, a method is provided for increasing oxygen affinity of hemoglobin S in a subject, the method comprising administering to a subject in need thereof a therapeutically effective amount of a crystalline solvaté of Compound 1.

[0101] In another aspect, a method is provided for treating oxygen deficiency associated with sickle cell anémia, the method comprising administering to a subject in need thereof a therapeutically effective amount of a crystalline solvaté of Compound 1.

EXAMPLES [0102] The following examples describe the préparation, characterization, and properties of the free base of Compound 1 Form I ansolvate. Unless otherwise stated, ail températures are in degrees Celcius (°C) and the following abbreviations hâve the following définitions:

DSC	Differential scanning calorimetry
DVS	Dynamic vapor sorption
HPLC	High performance liquid chromatography
NA	Not applicable
ND	Not determined
Q	Percent dissolved per unit time
RH	Relative humidity

Residual standard déviation

RRT Relative rétention time

SS-NMR Solid state nuclear magnetic résonance

TGA Thermogravimetric analysis

TG-IR Thermogravimetric infra red analysis

XRPD X-ray powder diffraction

VT-XRPD V ariable température X-ray powder diffraction

Synthetic Routes for Preparing Compound 1 [0103] The compound of formula (I) was synthesized as schematically described below and elaborated thereafter.

1.1 HDMLOA. R:BuU.5:)Hl .2 h.-10'3 ? 1h,rt!:aw

O'S ctt - '('Οι, 2516 2516; 2012

OH	haH	OMOV
A	MOMCl -	cS
	ather/ÜMI
11	9 2'K	12

MOMCl

QMQt/ .X_.Br

J i i k:imlda, r:b-li. s Meic-u^Me π ? -lire i ? -'trc: 7 h. -o-γ.: -rfc ?

1.3 Q'C: I h. 0^rC

1/ R:FCI,S:H2O

34¹½

Ju,ii yl uf Oiuanii. Clu-mifliy, 74(11), 4-311-4317, 2009

OH

F>U- i, DfVF	OMOM .X --CHO il T	125 HCl	OH .L. CHO (T
73 ta β ÜC	T -Ά ΌΜΟΜ	THF
94¾	13	81%	16

HCl

K2CO3

Example 1: Synthesis of Compound 15

DMF, 65C

81%

DIPEA

MOMCl

90%

OMOM

[0104] To a solution of 2-bromobenzene-l,3-diol (5 g, 26.45 mmol) in DCM (50 ml) at 0 °C was added DIPEA (11.54 mL, 66.13 mmol) and MOMCl (4.42 mL, 58.19 mmol). The mixture was stirred at 0 °C for 1.5 h, and then warmed to room température. The solution was diluted with DCM, washed with sat. NaHCC>3, brine, dried and concentrated to give crude product, which was purified by column (hexanes/EtOAc=4:l) to give desired product 15.58 g (90%).

OMOM

vjiviUM

Example 2: Synthesis of Compound 13 from 15

OMOM

Br BuLi, DMF

OMOM -78 to 0 °C

94% [0105] To a solution of 2-bromo-l,3-bis(methoxymethoxy)benzene (15) (19.9g, 71.8 mmol) in THF (150 mL) at -78 °C was added BuLi (2.5 M, 31.6 mL, 79.0 mmol) dropwise. The solution was stirred at -78 °C for 25 min (resulting white cloudy mixture), then it was warmed to 0 °C and stirred for 25 min. The reaction mixture slowly turns homogenous. To the solution was added DMF at 0 °C. After 25 min, HPLC showed reaction completed. The mixture was quenched with sat. NH4C1 (150 mL), diluted with ether (300 mL). The organic layer was separated, aq layer was further extracted with ether (2X200 mL), and organic layer was combined, washed with brine, dried and concentrated to give crude product, which was triturated to give 14.6 g desired product. The filtrate was then concentrated and purified by column to give additional 0.7 g, total mass is 15.3 g.

Example 3: Synthesis of Compound 13 from resorcinol 11

1.1 R.TMEDA, R:BuLi, S:THF, 2 h, -10’C

M 1.2 1 h, 0C; 83% OMOM

OrgLett 14(10), 2516-2519; 2012 ₀R: Λ-CHO

OMOM 1.1 RiTMEDA, R:BuLi, S:Me(CH2)4Me, rt ?-10°C ^<0MOM

1.2 -10°C; 2 h,-10”C;-10”C ? 0°C

1.3 0°C; 1 h, 0°C 13

1.4 R:HCI, S:H2O

84%

Journal of Organic Chemistry, 74(11), 4311-4317; 2009 [0106] A three-necked round-bottom flask equipped with mechanical stirrer was charged with 0.22 mol of NaH (50 % suspension in minerai oil) under nitrogen atmosphère. NaH was washed with 2 portions (100 mL) of n-hexane and then with 300 mL of dry diethyl ether; then 80 mL of anhydrous DMF was added. Then 0.09 mol of resorcinol 11, dissolved in 100 mL of diethyl ether was added dropwise and the mixture was left under stirring at rt for 30 min. Then 0.18 mol of MOMC1 was slowly added. After 1 h under stirring at rt, 250 mL of water was added and the organic layer was extracted with diethyl ether. The extracts were washed with brine, dried (Na₂SO4), then concentrated to give the crude product that was purified by silica gel chromatography to give compound 12 (93 % yield).

[0107] A three-necked round-bottom flask was charged with 110 mL of n-hexane, 0.79 mol of

BuLi and 9.4 mL of tetramethylethylendiamine (TMEDA) under nitrogen atmosphère. The mixture was cooled at -10 °C and 0.079 mol of bis-phenyl ether 12 was slowly added. The resulting mixture was left under magnetic stirring at -10 °C for 2 h. Then the température was raised to 0 °C and 0.067 mol of DMF was added dropwise. After 1 h, aqueous HCl was added until the pH was acidic; the mixture was then extracted with ethyl ether. The combined extracts were washed with brine, dried (Na₂SO4), and concentrated to give aldéhyde 13 (84%).

[0108] 2,6-bis(methoxymethoxy)benzaldehyde (13): mp 58-59 °C (n-hexane) ; IR (KBr) n: 1685 (C=O) cm¹; ’H-NMR (400 MHz, CDC1₃) δ 3.51 (s, 6H, 2 OCH₃), 5.28 (s, 4H, 2 OCH₂O), 6.84 (d, 2H, J = 8.40 Hz, H-3, H-5), 7.41 (t, IH, J = 8.40 Hz, H-4), 10.55 (s, IH, CHO); MS, m/e (relative intensity) 226 (M+, 3), 180 (4), 164 (14), 122 (2), 92 (2), 45 (100); Anal. Calc’d. for C11H14O5: C,58.40; H, 6.24. Found: C, 57.98; H, 6.20.

Example 4: The Synthesis of Compound 16

OMOM OH

[0109] To a solution of 2,6-bis(methoxymethoxy)benzaldehyde (13) (15.3 g, 67.6 mmol) in THF (105 mL) (solvent was purged with N₂) was added conc. HCl (12N, 7 mL) under N₂, then it was further stirred under N₂ for 1.5 h. To the solution was added brine (100 mL) and ether (150 ml). The organic layer was separated and the aqueous layer was further extracted with ether (2x200 mL). The organic layer was combined, washed with brine, dried and concentrated to give crude product, which was purified by column (300g, hexanes/EtOAc=85:15) to give desired product 16 (9.9 g) as yellow liquid.

Example 5: Synthesis of Compound 17

OH

CHO

OMOM

K2CO3

DMF °C

81%

[0110] To a solution of 2-hydroxy-6-(methoxymethoxy)benzaldehyde (16) (10.88 g, 59.72 mmol) in DMF (120 mL) (DMF solution was purged with N₂ for 10 min) was added K₂CO₃ (32.05 g, 231.92 mmol) and 3-(chloromethyl)-2-(l-isopropyl-lH-pyrazol-5-yl)pyridine hydrochloride (10) (15.78 g, 57.98 mmol). The mixture was heated at 65 °C for 1.5 h, cooled to rt, poured mto ice water (800 mL). The precipitated solids were isolated by filtration, dried and concentrated to give desired product (17, 18 g).

Example 6: Synthesis of Compound (I)

[OUI] To a solution of 2-((2-(l-isopropyl-lH-pyrazol-5-yl)pyridin-3-yl)methoxy)-6(methoxymethoxy)benzaldehyde (17) (18 g, 47.19 mmol) in THF (135 mL, solution was purged with N₂) was added conc. HCl (12N, 20 mL). The solution was stirred at rt for 3 h when HPLC showed the reaction complété. The mixture was added to a solution of NaHCCL (15 g) in water (1.2 L), and the resulting precipitate was collected by filtration, dried to give crude solid, which was further purified by column (DCM/EtOAc=60:40) to give pure product (15.3 g).

Example 7: Synthesis of Compound I (free base) and its HCl sait form [0112] Compound (I) free base (40g) was obtained from the coupling of the alcohol intermediate 7 and 2,6-dihydroxybenzaldedhye 9 under Mitsunobu conditions. A procedure is also provided below:

Example 8: Synthesis of Compound (I) by Mitsunobu coupling [0113] Into a 2000-mL three neck round-bottom flask, which was purged and maintained with an inert atmosphère of nitrogen, was placed a solution of [2-[l-(propan-2-yl)-lH-pyrazol-5yl]pyridin-3-yl]methanol (7) (70 g, 322.18 mmol, 1.00 equiv) in tetrahydrofuran (1000 mL). 2,6Dihydroxybenzaldehyde (9) (49.2 g, 356.21 mmol, 1.10 equiv) and PPI13 (101 g, 385.07 mmol, 1.20 equiv) were added to the reaction mixture. This was followed by the addition of a solution of DIAD (78.1 g, 386.23 mmol, 1.20 equiv) in tetrahydrofuran (200 ml) dropwise with stirring. The resulting solution was stirred ovemight at room température. The resulting solution was diluted with 500 ml of H₂O. The resulting solution was extracted with 3x500 ml of dichloromethane and the combined organic layers were dried over sodium sulfate and concentrated under vacuum. The residue was applied onto a silica gel column with EA:PE (1:501:3) as eluent to yield the crude product. The crude product was re-crystallized from ipropanol/H₂O in the ratio of 1/1.5. This resulted in 40 g (37%) of 2-hydroxy-6-([2-[l-(propan-2yl)-lH-pyrazol-5-yl]pyridin-3-yl]methoxy)benzaldehyde as a light yellow solid. The compound exhibited a melting point of 80-82 °C. MS (ES, m/z): 338.1 [M+l]. Ή NMR (300 MHz, DMSO-d6) δ 11.72(s, IH), 10.21(s, IH), 8.76(d, J=3.6Hz, IH), 8.24(d, J=2.7Hz, lH),7.55(m, 3H), 6.55(m,3H) ,5.21 (s, 2H), 4.65 (m, IH), 1.37 (d, J=5.1Hz, 6H). Ή NMR (400 MHz, CDC1₃) δ 11.96 (s, IH), 10.40 (s, IH), 8.77 (dd, 7 = 4.8,1.5 Hz, IH), 8.00 (d, 7= 7.8 Hz, IH),

7.63 (d, 7= 1.8 Hz, IH), 7.49 - 7.34 (m, 2H), 6.59 (d, 7= 8.5 Hz, IH), 6.37 (d, 7= 1.8 Hz, IH), 6.29 (d, 7= 8.2 Hz, IH), 5.10 (s, 2H), 4.67 (sep, 7= 6.7 Hz, IH), 1.50 (d, 7= 6.6 Hz, 6H).

[0114] In another approach, multiple batches of Compound (I) free base are prepared in multi gram quantifies (20g). The advantage of this route is the use of mono-protected 2,6dihydroxybenzaldehyde (16), which effectively éliminâtes the possibility of bis-alkylation side product. The mono-MOM ether of 2,6-dihydroxybenzaldehyde (16) can be obtained from two starting points, bromoresorcinol (14) or resorcinol (11) [procedures described in the Journal of Organic Chemistry, 74(11), 4311-4317; 2009 ]. Ail steps and procedures are provided below. Due to the presence of phenolic aldéhyde group, précautions (i.e., carry out ail reactions under inert gas such as nitrogen) should be taken to avoid oxidation of the phénol and/or aldéhyde group.

Préparation of compound I HCl sait: A solution of compound I (55.79 g, 165.55 mmol) in acetonitrile (275 mL) was flushed with nitrogen for 10 min, then to this solution was added 3N aqueous HCl (62 mL) at room température. The mixture was stirred for additional 10 min after the addition, most of the acetonitrile (about 200 mL) was then removed by évaporation on a rotary evaporator at around 32 °C, the remaining solution was frozen by cooling in an acetonedry ice bath and lyophilized to afford compound I HCl sait (59.4 g).

Example 9: Characterization of the HCl sait of Compound 1

Technique

Details

Resuit

XRPD	indexed	HCl sait of Compound 1
Microscope	-	pale yellow solids, thin blades/tablets, biréfringent
‘H NMR	DMSO-d6	consistent with structure, <0.01 moles MEK

XRPD	-	HCl sait of Compound 1
DVS	-	0.03% gain upon équilibration at 5% RH 0.10% gain from 5 to 95% RH 0.09% loss from 95 to 5%RH
post XRPD	HCl I + Free Base Form I

Example 10: Physical Stability of the HCl sait of Compound 1 Exposed to Water

Condition	Time (ail times are approximated)	Observation	XRPD Resuit
contacted w/ water	-	sheet formation after 5 min	-
water slurry	about 5 min	Floating yellow solids convert to white solids upon isolation	Free base (FB) I (indexed)
vacuum dried	about 1 day	Remain	FB I
water slurry	about 6 days	white, thin blades, biréfringent (B)	FB I + FB II

Example 11: Physical Stability of the HCl sait of Compound 1 with Grinding

Condition	Time	Observation	XRPD Resuit
grinding, dry	30 min	off white/pale yellow	HC1I
grinding, wet	30 min	off white/pale yellow paste	HC1I + FB I

Example 12: Physical Stability of the HCl sait of Compound 1 Exposed to Elevated

Température and/or Vacuum

Condition	Time	Observation	XRPD Resuit
RT vacuum	6 days	pale yellow, blades/plates, B	HCl I + FB I
30 °C	6 hrs	pale yellow, blades/tablets, B	HC1I
12 hrs	pale yellow, blades/tablets, B	HCl I + FB I
24 hrs	pale yellow, blades/tablets, B	HCl I + FB I
40 °C	6 hrs	pale yellow, blades/tablets, B	HCl I + FB I
12 hrs	pale yellow, blades/tablets, B	HCl I + FB I
24 hrs	pale yellow, blades/tablets, B	HCl I + FB I
40 °C vacuum	6 hrs	pale yellow, blades/tablets, B	HCl I + FB I
12 hrs	pale yellow, blades/tablets, B	HCl I + FB I
24 hrs	pale yellow, blades/tablets, B	HCl I + FB I
60 °C	6 days	pale yellow blades, B	HCl I + FB I
60 °C vacuum	6 days	pale yellow, blades, B; irregular residue	HCl I + FB I + other free base form
100 to 125 °C	20 min	pH paper above sample indicate acidic volatiles	HCl I + FB I + other free base form

Example 13: Génération of the Free Base of Compound 1 from the Disproportionation of the HCl sait of Compound 1 in Water (The starting material is the HCl sait of Compound 1).

Method	Observation	XRPD Resuit
1. contacted with water 2. sonicated 3. filtered and rinsed with water 4. dried under N2 for 10 minutes 5. vacuum RT, ovemight	1. pale yellow, wets poorly 2. white 3. - 4. - 5. -	FB I
1. contacted with water 2. sonicated for 5 minutes 3. slurried for 10 minutes 4. filtered, rinsed with water	1,- 2. pale yellow, turned white 3, -	FB I + other free base form

5. dried under N2 for 10 minutes 6. vacuum RT, ovemight 7. stored in freezer	4. - 5. white 6. - 7,-
1. slurry in water, RT, 8 days; seeded with FB II	1. thick white slurry
2. filtered, rinsed with water	2,-	FB II
3. vacuum RT, ovemight	3,-
2. sub sample of slurry	2,-	FB II (indexed)
3. rinsed with water	3.-

Example 14: Characterization Form I of the Free Base of Compound 1

Technique	Details	Resuit
XRPD	indexed	Free Base Form I

XRPD	-	Free Base Form I
TGA	25 to 350 °C	0.2% weight loss up to 100 °C
DSC	25 to 350 °C	endothermie event with onset near 97 °C
	22.7 °C	initial, fines, biréfringent
	91.2 °C	increase in particle size and biréfringence
	94.2 °C	increase in particle size and biréfringence
	95.7 °C	melt onset, larger particles from initial heating
	96.1 °C	melt continuation
	96.3 °C	melt complété, no crystallization upon melting
Hot Stage Microscopy	68.7 °C	fresh préparation, larger magnification
91.1 °C	increase in biréfringence
	94.8 °C	melt onset, larger particles, biréfringent
	95.4 °C	melt continuation
	95.9 °C	only few crystals remain, cooled to 92.6 °C
	92.6 °C	held for 2 to 3 minutes crystal growth to larger blades,began heating
	96.3 °C	complété melt

Ή NMR	DMSO-d6	consistent with structure
DVS	-	0.02% loss upon équilibration at 5% RH 0.22% gain from 5 to 95% RH 0.22% loss from 95 to 5%RH
post XRPD	Free Base Form I + other Free Base Material

Example 15: Characterization of Form II of the Free Base of Compound 1

Technique	Details	Resuit
XRPD	indexed	Free Base Form II

XRPD	initial	Free Base Form II
after 7 days	Free Base Form II
TGA	25 to 350 °C	0.1% weight loss up to 100 °C
DSC	25 to 350 °C	endothermie event with onset near 97 °C
‘H NMR	DMSO-d6	consistent with structure

Example 16: Characterization of Material N of the Free Base of Compound 1

Technique	Details	Resuit
XRPD	-	Free Base Material N
TGA	25 to 350 °C	0.2% weight loss up to 100 °C
DSC	25 to 350 °C	endothermie event with onset near 94 °C
‘H NMR	DMSO-d6	consistent with structure, no residual reaction solvent observed

Example 17: Compétitive Interconversion Slurries Between Free Base Forms I and II

Conditions

Solvent

Observation

XRPD Resuit

6 °C, 6 days	water	white	FBII
6 °C, 6 days	heptane	white	FB II
6 °C, 6 days	IPE	faint pale yellow	FBN

RT, 6 days	water	white	FB II
RT, 6 days	heptane	off white	FBII
RT, 6 days	IPE	pale yellow	FB N ’'rror‘ ^°°kmark not detïned.
RT, 6 days	toluene	pale yellow	FBN

57 °C, 2 days	water	fines, off white, B	FB II + FB I
57 °C, ovemight	heptane	blades and tablets, B	FB II
57 °C, ovemight	IPE	blades, laminated, pale yellow, B	FB II

Example 18: Compétitive Interconversion Slurries between Free Base Form II and Material N

Conditions

35 °C, 3 days

heptane

pale yellow fines, B

FB N

57 °C, 3 days

heptane

larger blades, and rosettes of blades, B

FB II

Example 19: Selected Experimental Methods [0115] Indexing: XRPD patterns are indexed by using proprietary SSCI software. Agreement between the allowed peak positions, marked with red bars within the figures, and the observed 10 peaks indicates a consistent unit cell détermination. Indexing and structure refinement are computational studies which are performed under the Procedures for SSCI Non-cGMP Activities. To confirm the tentative indexing solution, the molecular packing motifs within the crystallographic unit cells must be determined. No attempts at molecular packing were performed.

[0116] Differential Scanning Calorimetry (DSC): DSC was performed using a TA Instruments

Q2000 differential scanning calorimeter. Température calibration was performed using NISTtraceable indium métal. The sample was placed into an aluminum DSC pan, covered with a lid, and the weight was accurately recorded. A weighed aluminum pan configured as the sample pan was placed on the reference side of the cell. The data acquisition parameters and pan configuration for each thermogram are displayed in the image in the Data section of this report. The method code on the thermogram is an abbreviation for the start and end température as well as the heating rate; e.g., -30-250-10 means from -30 °C to 250 °C, at 10 °C/min. The following summarizes the abbreviations used in each image for pan configurations: Tzero crimped pan (TOC); and Lid not crimped (NC).

[0117] Dynamic Vapor Sorption (DVS): Dynamic vapor sorption (DVS) data were collected on a VTI SGA-100 Vapor Sorption Analyzer. NaCl and PVP were used as calibration standards. Samples were not dried prior to analysis. Adsorption and desorption data were collected over a range from 5 to 95% RH at 10% RH incréments under a nitrogen purge. The equilibrium criterion used for analysis was less than 0.0100% weight change in 5 minutes with a maximum équilibration time of 3 hours. Data were not corrected for the initial moisture content of the samples.

Microscopy [0118] Hot Stage Microscopy: Hot stage microscopy was performed using a Linkam hot stage (FTIR 600) mounted on a Leica DM LP microscope equipped with a SPOT Insight™ color digital caméra. Température calibrations were performed using USP melting point standards. Samples were placed on a cover glass, and a second cover glass was placed on top of the sample. As the stage was heated, each sample was visually observed using a 20x 0.40 N. A. long working distance objective with crossed polarizers and a first order red compensator. Images were captured using SPOT software (v. 4.5.9).

[0119] Polarized Light Microscopy: During the course of expérimentation generated samples were viewed utilizing a microscope with cross polarized light to observe morphology and biréfringence. Samples were visually observed at 40x magnification.

'H Solution Nuclear Magnetic Résonance (*H NMR) [0120] SSCI: Samples were prepared for NMR spectroscopy as -5-50 mg solutions in the appropriate deuterated solvent. The spécifie acquisition parameters are listed on the plot of the first full spectrum of each sample in the data section for samples run at SSCI.

[0121] Spectral Data Solutions: For samples run using Spectral Data Solutions (subcontractor), the solution Ή NMR spectra were acquired at ambient température on a Varian ^yA,/rrINOVA-400 spectrometer ^H Larmor Frequency = 399.8 MHz). The spécifie acquisition parameters are listed on the spectral data sheet and on each data plot of the spectrum of the sample.

Thermogravimetric Analysis (TGA) [0122] TG analyses were performed using a TA Instruments 2950 thermogravimetric analyzer. Température calibration was performed using nickel and Alumel™. Each sample was placed in an aluminum pan and inserted into the TG fumace. The fumace was heated under a nitrogen purge. The data acquisition parameters are displayed above each thermogram in the Data section 15 of this report. The method code on the thermogram is an abbreviation for the start and end température as well as the heating rate; e.g., 25-350-10 means from 25 °C to 350 °C, at 10 °C/min. The use of 0 as the initial température indicates sample run initiated from ambient.

XRPD Analysis [0123] TNEL: XRPD patterns were collected with an Inel XRG-3000 diffractometer. An incident beam of Cu Ka radiation was produced using a fine-focus tube and a parabolically graded multilayer mirror. Prior to the analysis, a silicon standard (NIST SRM 640d) was analyzed to verify the Si 111 peak position. A specimen of the sample was packed into a thinwalled glass capillary, and a beam-stop was used to minimize the background from air. Diffraction patterns were collected in transmission geometry using Windif v. 6.6 software and a curved position-sensitive Equinox detector with a 2Θ range of 120°. The data-acquisition parameters for each pattern are displayed above the image in the Data section of this report. [0124] PANalytical Transmission: XRPD patterns were collected with a PANalytical X'Pert PRO MPD diffractometer using an incident beam of Cu radiation produced using an Optix long, fine-focus source. An elliptically graded multilayer mirror was used to focus Cu Ka X-rays through the specimen and onto the detector. Prior to the analysis, a silicon specimen (NIST SRM 640d) was analyzed to verify the Si 111 peak position. A specimen of the sample was sandwiched between 3 pm thick films and analyzed in transmission geometry. A beam-stop, short antiscatter extension, and an antiscatter knife edge were used to minimize the background

generated by air. Soller slits for the incident and diffracted beams were used to minimize broadening from axial divergence. Diffraction patterns were collected using a scanning positionsensitive detector (X'Celerator) located 240 mm from the specimen and Data Collecter software 5 v. 2.2b. The data-acquisition parameters for each pattern are displayed above the image in the

Data section of this report including the divergence sût (DS) before the mirror and the incidentbeam antiscatter slit (SS).

[0125] PANalytical Reflection: XRPD patterns were collected with a PANalytical X'Pert PRO MPD diffractometer using an incident beam of Cu Kot radiation produced using a long, fine10 focus source and a nickel filter. The diffractometer was configured using the symmetric BraggBrentano geometry. Prior to the analysis, a silicon specimen (NIST SRM 640d) was analyzed to verify the observed position of the Si 111 peak is consistent with the NIST-certified position. A specimen of the sample was prepared as a thin, circular layer centered on a silicon zerobackground substrate. Antiscatter slits (SS) were used to minimize the background generated by 15 air. Soller slits for the incident and diffracted beams were used to minimize broadening from axial divergence. Diffraction patterns were collected using a scanning position-sensitive detector (X'Celerator) located 240 mm from the sample and Data Collecter software v. 2.2b. The data acquisition parameters for each pattern are displayed above the image in the Data section of this report including the divergence slit (DS) and the incident-beam SS.

[0126] Approximate Solubility: A weighed sample was treated with aliquots of the test solvent at room température. The mixture was sonicated between additions to facilitate dissolution. Complété dissolution of the test material was determined by Visual inspection. Solubility was estimated based on the total solvent used to provide complété dissolution. Some samples were then heated and observed visually for complété dissolution. The actual solubility may be greater than the value calculated because of the use of solvent aliquots that were too large or due to a slow rate of dissolution. The solubility is expressed as “less than” if dissolution did not occur during the experiment. If complété dissolution was achieved as a resuit of only one aliquot addition, the solubility is expressed as “greater than”.

[0127] Anti-Solvent Additions: Compound 1/organic solvent solutions were contacted with 30 solvents that Compound 1 was determined to be poorly soluble or insoluble in. These anti solvent additions were added to help lower the solubility of the solvent System and induce crystallization.

[0128]

Cooling and Slow Cools: Solutions were prepared in the selected solvent or solvent/anti-solvent system. These solutions were chilled below room température within a refrigerator for varying lengths of time in an attempt to induce nucléation. The presence or absence of solids was noted. Upon observation of solids, in quantities sufficient for analysis, isolation of material was conduction. If insufficient quantities were présent further cooling was performed in a freezer. Samples were either isolated for analysis wet or as dry powders.

[0129] Compression: Selected samples were compressed utilizing a KBr die and a Carver hydraulic press. An applied load of 10000 Ibs was applied to the die shaft for approximately 20 minutes.

[0130] Crystallization from Solution: Saturated solutions were generated at ambient and then capped. Nucléation was observed to occur from these Systems during évaluation of the Free Base of Compound 1.

[0131] Fast Evaporation: Solutions were prepared in selected solvents and agitated between aliquot additions to assist in dissolution. Once a mixture reached complété dissolution, as judged 15 by visual observation, the solution was allowed to evaporate at ambient température in an uncapped vial or at ambient under nitrogen. The solids that formed were isolated for évaluation.

[0132] Milling: Selected material was milled utilizing a Reitch Mill. The material was loaded into an agate lined milling vessel followed by the addition of an agate bail. The vessel was then placed on to the mill and milled for approximately 30 minutes at frequency of 1/30 seconds. The 20 milling was stopped approximately every 10 minutes and material scraped from the wall before further milling.

[0133] Slurry: Solutions were prepared by adding enough solids to a given solvent so that excess solids were présent. The mixture was then agitated in a sealed vial at either ambient or an elevated température. After a given amount of time, the solids were isolated for analysis.

[0134] Température and Relative Humidity Stress: Selected materials were stressed at elevated related humidity and/or température. Relative humidity jars (saturated sait solutions used to generate desired relative humidity) were utilized to store selected samples. The following relative humidity jars were utilized during évaluation: 75% RH (NaCI) and 60% (NaBr), to investigate the effects of humidity. Températures utilized were ambient, 30,40, 60, and 100-125 °C.

[0135] Vacuum: Selected materials were stressed under reduced pressure for a set time period. Initial stressing was conducted with the in-house vacuum system with absolute pressure readings <500 mTorr, typically 30 to 50 mTorr (0.030 to 0.05 mm Hg). Additional vacuum stressing was conducted at 48 mmHg utilizing a portable lab vacuum and bleed to simulate conditions similar to those expected during process.

Example 20: Disproportionation of the HCl sait [0136] The disproportionation of the HCl sait in water was utilized to generate free base. The nucléation of Free Base Form I occurs first. Extending the slurry time induces the transformation to a more thermodynamically stable phase relative to Form I, Free Base Form II.

[0137] Three anhydrous materials of the free base were identified; Free Base Forms I, II, and Material N. Free Base Material N appears to be most stable form, relative to Forms I and II, at room température. Free Base Material N is enantiotropic relative to Form II, and will transform reversibly at a spécifie transition température (estimated near 42 °C). Above the transition température, Free Base Form II appears to be the most stable form, relative to Form I and Material N.

[0138] The HCl sait (termed “HCl Form I”) was subjected to various stress conditions and monitored by XRPD to evaluate physical stability. As discussed, disproportionation occurred during the DVS experiment of the HCl sait, indicating instability upon exposure to elevated humidity. Disproportionation is further évident with wet milling or in direct contact with water (e.g. slurry) as shown by the presence of Free Base Forms I or II, identified by XRPD. The volatilization and loss of HCl upon heating and/or vacuum is shown by the presence of Free Base Form I, identified by XRPD, and also indicates instability at these conditions.

• Contact with water resulted in a visual color change of the material from pale yellow to white; physical changes were also observed microscopically. Immédiate disproportionation occurs. XRPD analysis identified the resulting material from a water slurry (~5 minutes) as Free Base Form I. Free Base Form II also becomes évident if the amount of time in the slurry is extended.

• The volatilization of HCl was évident within hours of exposure to drying conditions. Conversion to Free Base Form I was observed by XRPD at 30 °C (after 12 hrs), 40 °C (after 6 hrs), and at 40 °C/48 mmHg (after 6 hrs).

• Free Base Material C becomes évident at more extreme conditions involving elevated températures. Heating HCl Form I up to 125 °C induces the loss of acidic volatiles (judged visually by use of pH paper held above sample). XRPD analysis identifies the resulting sample as a mixture of HCl Form I, Free Base Form I, and Free Base Material

C. Exposing the HCl sait to 60 °C under vacuum for 6 days provides the same resuit. The nature of Material C is not established [0139] The HCl sait was shown to disproportionate immediately in water. This phenomenon was utilized to generate free base. The nucléation of Free Base Form I occurs first. Extending the slurry time induces the transformation to a more thermodynamically stable phase relative to Form I, Free Base Form IL • A 20 ml vial was charged with 266.4 mg of HCl Form I and contacted with 10 ml of water. The sample was sonicated until the pale yellow material changed color to white. The resulting solids were collected by filtration (water aspirated) and rinsed with 10 ml of water. A nitrogen purge was blown over the sample for approximately 10 minutes prior to exposure to vacuum at ambient température to dry ovemight. The resulting material was analyzed by XRPD and determined to be Free Base Form I.

• A 250 ml Erlenmeyer flask was charged with 6.0250 grams of HCl Form I and contacted with 220 mL of water. The sample was sonicated for approximately 5 minutes to disperse the material. The yellow material changed color to white during sonication. A stir bar was added and the sample was stirred at 700 RPM for approximately 10 minutes. The solids were collected by filtration and rinsed with 220 ml of water followed by a nitrogen purge over the sample for approximately 10 minutes prior to exposure to vacuum at ambient température. The sample was dried at this condition for approximately 24 hours yielding 5.1834 grams of material. The resulting material was analyzed by XRPD and determined to be a mixture of Free Base Form I and Free Base Material D. (The nature of Material D is not established.) [0140] The procedure used to generate Free Base Form II is described below.

• A 20 ml vial was charged with 477.5 mg of HCl Form I lot 20 and contacted with 20 ml of water. The sample was sonicated until the pale yellow material changed color to white. A small amount of sample (mixture of Free Base Forms I and II) was added as seeds. A stir bar was added and the sample was stirred at 200 RPM for 8 days. The resulting solids were collected by filtration (water aspirated) and rinsed with 15 ml of water. The sample was exposed to vacuum at ambient température to dry ovemight. The resulting material was analyzed by XRPD and determined to be Free Base Form IL

Example 21: Additional Procedures for the Préparation of the free base of Form I, Form

II, and From N

Conversion of the free base of Compound 1 to the HCl sait [0141] General Procedure: Slowly treat a solution of the free base of Compound 1 in MEK (5 5 vol) with conc HCl (1.5 eq). Cool the resulting slurry to 0-5 °C for 1 h and filter. Wash solids with MEK (1 vol). Dry under vacuum at 30-35 °C.

[0142] Préparation A: Following the general procedure above, 35 g of crude Compound 1 was processed to provide the HCl sait as a pale yellow solid (32.4 g, 82% yield, 99.8% purity by HPLC).

Préparation of the free base Form Ifrom the HCl Sait of Compound 1 [0143] General Procedure: Vigorously stir a slurry of the HCl sait of Compound 1 in DIW (10 vol) for 5 min to 2 h. Filter the slurry, wash with DIW (2x1 vol), dry on funnel, then further dry under vacuum at 30-35 °C.

[0144] Préparation A: Following the general procedure above, after stirring for lh, 32 g of the 15 HCl sait of Compound 1 was processed to provide the free base as a.pale yellow solid (27.3 g,

95% yield, 99.8% purity by HPLC; DSC indicates Form I).

[0145] Préparation B: Following the general procedure above, after stirring for lh, 39 g of the HCl sait of Compound 1 was processed to provide the free base as a pale yellow solid (31.8 g, 90% yield, >99.9% purity by HPLC)).

[0146] Préparation C: Thus, the HCl sait of Compound 1 (134 g) was vigorously stirred in water (10 vol) until the material appeared as a finely dispersed white slurry. After filtration and drying, a white crystalline solid (116 g, 96% recovery, >99.9% purity by HPLC) was isolated.

[0147] Préparation D: The purpose of this experiment was to préparé the free base from Compound 1, HCL Thus, the HCl sait of Compound 1 (65.3 g) was vigorously stirred in water 25 (10 vol) until the material appeared as a finely dispersed white slurry. After filtration and drying, a white crystalline solid (57.5 g, 97.6% recovery, >99.9% purity by HPLC) was isolated.

Préparation of GBT000440free base Form IIfrom GBT000440free base Form I [0148] General Procedure: Stir a slurry of the free base of Compound 1 Form I in an appreciate solvent (e.g. heptane or water) (10 vol) for 1-7 days. Filter the slurry, wash with DIW 30 (2x1 vol), dry on funnel, then further dry under vacuum at 30-35 °C.

[0149] Préparation A: Thus, the free base of Compound 1, Form I (114 g) was stirred in nheptane (10 vol) at 35 °C. After 4 days, XRPD indicated the material was Form II. The slurry was filtered and dried to provide 110 g off white solid.

[0150] Préparation B: the free base of Compound 1 (5 g) was slurried in heptanes (10 vol 50mlL) at room température. After 4 days, the slurry was filtered to provide an off-white solid.

Préparation C: the free base of Compound 1 (5.8 kg) was slurried in heptanes (10 vol) at room température. After 2 days, the slurry was filtered and washed with 2x2 vol n-heptane to provide 4.745 kg of Form II as an off-white solid.

[0151] Préparation D: the free base of Compound 1 (5 g) was slurried in water. After 4 days, the slurry was filtered to provide an off-white solïà.Preparation of GBT000440 free base Form Nfrom GBT000440free base Form I or Form II [0152] General Procedure: Stir a slurry of the free base of Compound 1, Form I in MTBE (4 vol) at room température for at least 4 days. After 4 days, filter the slurry to provide an off-white solid. Obtain XRPD to confirm polymorph as Material N.

[0153] Préparation A: Following the general procedure above, 27 g of the free base of Compound 1, Form I (48TRS079) was stirred in MTBE at 18-23 °C for 4 days. DSC indicated it should be Material N. Isolated 22.2 g cream colored solid (82% recovery, 99.9 purity by HPLC). XRPD analysis planned.

[0154] Préparation B: Following the general procedure above, 31 g of the free base of Compound 1, Form I was stirred in 3 vol MTBE at 18-23 °C for 4 days.

[0155] Préparation C: the free base of Compound 1, Form I (13KWBO23, 1 g) was slurried in MTBE (5 vol) at room température. Slurry was seeded with Material N (50 mg). After 4 days, the slurry was filtered to provide a off-white solid. DSC indicated the melting point was the same as Material N.

[0156] Préparation D: The purpose of this experiment was to couvert the the free base of Compound 1, Form II to Material N. Thus, the free base of Compound 1 (0.5 g) was stirred in 5 vol of di-n-propyl ether at 18-23 °C. After 2 days, DSC corresponded to the pattern observed for Material N. XRPD analysis confirmed Material N had been formed.

[0157] Préparation E: To the free base of Compound 1, Form II (5 g) was added diisopropyl ether (5 vol, 25 mL) at room température. After 4 days, the slurry was filtered to provide a offwhite solid. DSC indicates Material N.

Example 22: Preliminary Solvent-based Screens [0158] Rapid, solvent-based screens were conducted m an attempt to détermine the most stable form of the free base of Compound 1. The study also provides a preliminary assessment of the propensity of these materials to exist in various crystal forms. Generated solids were observed by polarized light microscopy (PLM) and/or analyzed by X-ray powder diffraction (XRPD), comparing the resulting XRPD patterns to known patterns of Compound 1.

[0159] If possible, XRPD patterns were indexed. Indexing is the process of determining the size and shape of the crystallographic unit cell given the peak positions in a diffraction pattern. The term gets its name from the assignaient of Miller index labels to individual peaks. XRPD indexing serves several purposes. If ail of the peaks in a pattern are indexed by a single unit cell, this is strong evidence that the sample contains a single crystalline phase. Given the indexing solution, the unit cell volume may be calculated directly and can be useful to détermine their solvation states. Indexing is also a robust description of a crystalline form and provides a concise summary of ail available peak positions for that phase at a particular thermodynamic state point.

[0160] Materials exhibiting unique crystalline XRPD patterns, based on visual inspection of peaks associated with these materials, were given letter désignations. The letter désignation is tentatively associated with the word 'Material' if insufficient characterization data is available. The nomenclature is used only to aid in the identification of unique XRPD patterns and does not imply that the stoichiometry, crystalline phase purity, or chemical purity of the material is known. Materials are further designated as forms with Roman numéral désignations (i.e., Free Base Material A = Free Base Form I), when phase purity (obtained through indexing of the XRPD pattern or single crystal structure élucidation) and chemical identity/purity (obtained through proton NMR spectroscopy) of the material is determined.

[0161] Three anhydrous materials were identified: Forms I, II, and Material N. Material N appears to be most stable form, relative to Forms I and II, at room température. Material N is enantiotropic relative to Form II, and will transform reversibly at a spécifie transition température (estimated near 42 °C). Above the transition température, Form II appears to be the most stable form, relative to Form I and Material N.

[0162] Materials C and D are used to identify a few additional, low intensity peaks observed in XRPD patterns which were predominantly composed of the Free Base Form I of Compound 1 or mixtures of the HCl Form I and Free Base Form I of Compound 1.

Example 23: Anhydrous Ansolvate Forms

Form I [0163] Free Base Form I is a metastable, anhydrous phase of the free base that is formed immediately from the disproportionation of the HCl sait in water. A représentative XRPD pattern of Form I was successfully indexed and the unit cell volume is consistent with anhydrous free base. Visual comparison of the XRPD pattern to the historical pattern of the free base provided indicates the material may be similar; however, the historical pattern appears to exhibit additional peaks from a potential mixture.

[0164] The ^lH NMR spectrum is consistent with the chemical structure of Compound 1. The chemical shift at approximately 2.5 ppm is assigned to DMSO (due to residual protons in the NMR solvent). Peaks that could be associated with residual solvents were not visible, consistent with the anhydrous unit cell volume determined from the indexing solution above and the negligible wt% loss observed by TGA discussed below.

[0165] Thermograms (TG) data shows negligible weight loss, 0.2%, up to 100 °C, consistent with an anhydrous form. The DSC exhibits a single endotherm with an onset near 97 °C (similar to what is observed for Form II). The endotherm is consistent with a melt by hot stage microscopy. However, changes in particle size and biréfringence were évident prior to the melt; a possible phase change occurred. Consequently, if a phase change occurred and an endotherm similar to that of Free Base Form II was observed, it can be inferred that the observed melt is truly not of Form I but of the resulting phase, most likely Form II.

[0166] The DVS isotherm indicates Form I is not hygroscopic. Negligible weight gain and loss, 0.2%, was observed through sorption/desorption. By XRPD, the material recovered from the DVS experiment was predominately Free Base Form I with a few additional peaks. The additional peaks were termed Free Base Material D. The nature of Material D is unknown; however, the appearance of another phase(s) indicates that Form I is not likely physically stable at elevated humidity conditions (at ambient température).

Form II [0167] Free Base Form II is an anhydrous phase of the free base. Form II is enantiotropically related to Material N, where it is the thermodynamically stable form above an estimated transition température of 42 °C. Form II can be generated in solvents that do not form known solvatés; such as heptane, IPE, MTBE, or toluene; through short-term slurry conversions of

Form I (where the crystallization kinetics delay the nucléation of the more stable form) or elevated température slurries (above 42 °C). A représentative XRPD pattern of Form II was successfully indexed and the unit cell volume is consistent with anhydrous free base of Compound 1.

[0168] The ’H NMR spectrum is consistent with the chemical structure of Compound 1. The chemical shift at approximately 2.5 ppm is assigned to DMSO (due to residual protons in the NMR solvent). Peaks that could be associated with residual solvents were not visible, consistent with the anhydrous unit cell volume determined from the indexing solution above and the negligible wt% loss observed by TGA discussed below.

[0169] Thermograms (TG) data show negligible weight loss, 0.1%, up to 100 °C, consistent with an anhydrous form. The DSC exhibits a single endotherm (80.1 J/g) with an onset near 97 °C.

[0170] Form II remained unchanged after 7 days at ambient storage, through reanalysis by XRPD. The form is known to be thermodynamically metastable, relative to Material N, at this condition; however, the kinetics of polymorph conversion may be slow at ambient conditions in the solid state.

Material N [0171] Free Base Material N is an anhydrous phase of the free base. Material N is enantiotropically related to Form II, where it is the thermodynamically stable form below an estimated transition température of 42 °C. Given the opportunity, Material N can be generated through slurries in solvents that do not form known solvatés; such as heptane, IPE, MTBE, or toluene; at températures below 42 °C. The following is an example of a laboratory scale procedure used to generate Free Base Material N.

• 53.0 mg of Free Base Form I was contacted with 2 ml of an IPE/free base solution (concentration 13 mg/ml). A stir bar was added and the sample was slurried for 7 days at ambient. The solution was decanted from the sample and the remaining solids briefly dried under nitrogen. Characterization Data indicates Material N is a unique crystalline phase.

[0172] The *H NMR spectrum is consistent with the chemical structure of Compound 1. The chemical shift at approximately 2.5 ppm is assigned to DMSO (due to residual protons in the

NMR solvent). Peaks that could be associated with residual solvents were not visible, consistent with the negligible wt% loss observed by TGA discussed below.

[0173]

Thermograms (TG) data show negligible weight loss, 0.2%, up to 100 °C, consistent

with an anhydrous form. The DSC exhibits a single endotherm (82.8 J/g) with an onset at 94 °C.

[0174] Tentative Détermination of the Thermodynamic Relationship between Free Base Forms

I, II, and Material N [0175] Characterization data indicates that Forms I, II, and Material N are unique crystalline phases; however, only the XRPD patterns of Forms I and II were successfully indexed to confirm phase purity. Therefore, any proposed thermodynamic relationship between these materials is a working hypothesis, where the phase purity of Material N is assumed.

[0176] Phase transitions of solids can be thermodynamically réversible or irréversible.

Crystalline forms which transform reversibly at a spécifie transition température (T_p) are called enantiotropic polymorphs. If the crystalline forms are not interconvertable under these conditions, the system is monotropic (one thermodynamically stable form). Several rules hâve been developed to predict the relative thermodynamic stability of polymorphs and whether the relationship between the polymorphs is enantiotropic or monotropic. The heat of fusion rule is applied within this study. The heat of fusion rule states that if the higher melting form has the lower heat of fusion then the two forms are enantiotropic, otherwise they are monotropic.

[0177] Material N appears to be most stable form, relative to Forms I and II, at room température. Based on the heats of fusion and melts determined by DSC, Material N is enantiotropic relative to Form II, and will transform reversibly at a spécifie transition température (T^77-77). Due to a possible phase change of Form I to Form II that occurred prior to the observed endotherm in the DSC, the relationship of Form I with either Material N or Form II cannot be conclusively determined through the heat of fusion rule. However, through various interconversion slurries, it was shown that Form I is the least thermodynamically stable form between 6 °C and A⁷⁷. In addition, assuming that Form I spontaneously converted to Form II in the DSC at elevated températures (prior to the observed melt), it must follow that Form II is also more stable than Form I above A⁷⁷.

Example 24: Estimated Transition Température [0178] The estimated transition température between two enantiotropically related forms can be calculated from their melt onsets and heats of fusion based on the équation shown below.

AHf 2 -AHf.i ⁺(Cp.Uq ~Cp.l)'(^Tf,l ~Tf,2)

ΔΗ_Γ,2

Tf,2

AHf.i

Tf,l ⁺ (Cp.liq £p.lVⁿ fjh (Tf.2

Where, (Cp.liq ~Cp.l) ⁼ and k = 0.005 [0179] Between Material N and Form II, the équation estimâtes a transition température of 42 °C. To summarize, the relative stability of the forms from most to least stable is shown below.

Température Range*	Relative Stability	Commente
Below 6 °C	N>II	Relationships to Form I are not established below this temp
Between 6 °C and jN-II	N > II > I
Above Y'11	(II > N) and (II >D	Relationship between Form I and Material N is not established above this temp

*Ί^Ν'^Π is estimated to be near 42 °C

Example 25: Energy - Température Diagram [0180] The Energy - Température Diagram of FIG. 17 is a semi-quantitative graphical solution of the Gibbs - Helmholtz équation, where the enthalpy (H) and free energy (G) isobars for each form are depicted as a function of température.

Example 26: Compétitive Interconversion Slurry Expérimente [0181] Interconversion experiments were performed to support the thermodynamic relationship between polymorphs illustrated by the Energy - Température Diagram above. Interconversion or compétitive slurry experiments are a solution-mediated process that provides a pathway for the less soluble (more stable) crystal to grow at the expense of the more soluble crystal form. Outside the formation of a solvaté or dégradation, the resulting more stable polymorph from an interconversion experiment is contemplated to be independent of the solvent used because the more thermodynamically stable polymorph has a lower energy and therefore lower solubility. The choice of solvent affects the kinetics of polymorph conversion and not the thermodynamic relationship between polymorphie forms.

[0182] The results of the interconversion studies are consistent with the tentative Energy Température Diagram shown herein. Binary slurries were prepared at ambient, 6, and 57 °C using Forms I and II. Form II resulted from the majority of these experiments, confirming that Form II is more stable relative to Form I within this température range.

[0183] A few of the experiments conducted at ambient and 6 °C resulted in Material N. Although this does not provide spécifie clarification between Forms I and II, it does provide evidence that Material N is the most stable form relative to both Forms I and II at these températures (which were conducted below the estimated transition température of 42 °C). Additional interconversion slurries between Form II and Material N were conducted at températures which bracket this estimated transition température and confirm that Form II and Material N are enantiotropically related.

Example 27: Solid-state Nuclear Magnetic Résonance [0184] ¹³C and ¹⁵N spectra acquired for the three polymorphie forms I, II and Material N. See

FIGS. 10 and 11. Spectra were acquired at 253K to prevent any low température transitions occurring during measurement and acquisition parameters optimised for each polymorphie form. [0185] Based on solid-state nuclear magnetic résonance, ail three forms are crystalline and are distinct polymorphie forms. Form I contains one molécule per asymmetric unit, Form II contains two molécules per asymmetric unit and Form N contains four molécules per asymmetric unit. See the ¹⁵N spectra in FIG. 11.

Example 28: Chemical and Physical Stability Evaluation of the Free Base Form I of Compound 1 [0186] A mixture predominately composed of Free Base Form I (with Free Base Material D) were exposed to stability conditions to assess physical and chemical stability. Three conditions were used; open to 25 °C / 60% RH, open to 40 °C / 75% RH, and closed to 60 °C. Physical stability was evaluated by XRPD. Chemical stability was determined through UPLC and ^JH NMR, when applicable. Materials were tested after 1, 7, and 14 days of exposure.

Chemical Stability of Free Base Form I [0187] For the free base stability sample, UPLC showed very low levels of impurities présent.

The level of impurities did not rise significantly after 14 days of âge. This would seem to indicate good chemical stability against the conditions used for stability assessment. The *H

NMR spectra of samples exposed to 60 °C (14 days) were also consistent with this conclusion.

Physical Stability of Free Base Form I [0188] The free base of Compound 1 remained unchanged, by XRPD, at 25 °C / 60% RH. However, différences were observed at the other two conditions. The few, minor peaks attributed to Free Base Material D were no longer observed, indicating that Material D is metastable and is not sustained at elevated températures. In addition, Free Base Form II was observed after 7 days of âge. This is consistent with the conclusions discussed herein, where Free Base Form II is more stable relative to Free Base Form I at these températures.

Example 29: Physical Stability Evaluation of the Free Base Form II and Material N (Form N) of Compound 1 [0189] DSC was modulated at low underlying heating rate, followed by X-ray powder diffraction. A low underlying heating rate was used of of 0.02 °C min'¹. The température was 80 °C for form N and 90 °C for form II. Exposure was essentially isothermal, covering a température range with sensitivity to detect changes in physical form. The résultant materials were examined by X-ray powder diffraction. No changes in physical form were observed for either polymorphie form II or polymorphie form N (i.e., material N).

[0190] Forms II and N were exposed to 40 °C/75% relative humidity (RH), 80 °C, 80 °C /80%RH for 9 days followed by X-ray powder diffraction. No changes in physical form were observed for either polymorphie form II or polymorphie form N.

[0191] The thermodynamic barrier for inter-conversion between polymorphie form II and form 20 N is high, and physical stability is good for both forms. Thermally induced inter-conversion between form II and form N is unlikely to occur.

Example 30: The Relative Thermodynamic Stability of Polymorphie Forms II and N.

[0192] Extended solvent mediated maturation studies were conducted with 1:1 w/w mixtures of polymorphie form II and form N. Hexane provided a good medium for solvent assessments.

The températures used include -20 °C, -10 °C, 0 °C, 10 °C, 20 °C, 30 °C, 40 °C and 50 °C. Significantly increased solubility was observed at 30 °C, 40 °C and 50 °C. Solids derived from maturation at -20 °C, -10 °C, 0 °C, 10 °C, 20 °C were analyzed by X-ray powder diffraction. In each case, significant conversion to Form N was observed.

[0193] Form N is thermodynamically more stable than form II at températures of 20 °C and lower. An enantiotropic relationship between the two forms is likely to exhibit équivalence in thermodynamic stability at ca. 30-40 °C .

Example 31: Morphology of Form N [0194] Initial examination of a batch of polymorphie form N indicates an acicular morphology.

[0195] While this invention has been described in conjunction with spécifie embodiments and examples, it will be apparent to a person of ordinary skill in the art, having regard to that skill 5 and this disclosure, that équivalents of the specifically disclosed materials and methods will also be applicable to this invention; and such équivalents are intended to be included within the following claims.

Claims (19)

1. Claims

   1. A crystalline ansolvate of Compound 1:
2. 2. The crystalline ansolvate of claim 1, which is substantially free of a solvated polymorph of Compound 1.
3. 3. A composition comprising the crystalline ansolvate of claim 1.
4. 4. The crystalline ansolvate of claim 1, wherein the crystalline ansolvate comprises one or more of Form I, Form II or Material N, wherein

   Form I is characterized by at least one X-ray powder diffraction peak (Cu Ka radiation) selected from 12.82°, 15.74°, 16.03°, 16.63°, 17.60°, 25.14°, 25.82° and 26.44° (each ±0.2 °20);

   Form II is characterized by at least one X-ray powder diffraction peak (Cu Ka radiation) selected from 13.37°, 14.37°, 19.95° and 23.92°2θ (each ±0.2 °2Q); and Material N is characterized by at least one X-ray powder diffraction peak (Cu Ka radiation) selected from 11.65°, 11.85°, 12.08°, 16.70°, 19.65° and 23.48 °20 (each ±0.2 °20).
5. 5. The crystalline ansolvate of claim 4, wherein the crystalline ansolvate comprises Form IL
6. 6. The crystalline ansolvate of claim 4, wherein the crystalline ansolvate comprises Material N.
7. 7. A method of preparing a crystalline ansolvate of claim 1 comprising slurrying a sait of Compound 1 in a solvent and disproportionating the Compound 1 sait to form the crystalline ansolave.
8. 8. Use of a therapeutically effective amount of a crystalline ansolvate of claim 1 for increasing oxygen affinity of hemoglobin S.
9. 9.

   Use of a therapeutically effective amount of a composition of claim 3 for increasing oxygen affînity of hemoglobin S.
10. 10. Use of a therapeutically effective amount of a crystalline ansolvate of claim 1 for treatment of oxygen deficiency associated with sickle cell anémia..
11. 11. Use of a therapeutically effective amount of a composition of claim 3 for treatment of oxygen deficiency associated with sickle cell anémia, the method comprising administering to a subject in need thereof.
12. 12. A crystalline solvaté of Compound 1 :

    ΌΗ

    Compound 1
13. 13. The crystalline solvaté of claim 12, which is substantially free of an ansolvated polymorph of Compound 1.
14. 14. A composition comprising the crystalline solvaté of claim 12 or claim 13.
15. 15. The crystalline solvaté of claim 12, comprising one or more of Material E, Material F, Material G, Material H, Material J, Material K, Material L, Material M, Material O or Material P.
16. 16. The crystalline solvaté of claim 15, wherein:

    Material E is characterized by at least one, two, or three X-ray powder diffraction peak (Cu Ka radiation) selected from 8.69,11.73, 12.10,15.26, 16.11, 17.45, 22.39, 22.55 and 23.70 ± 0.20;

    Material F is characterized by at least one, two, or three X-ray powder diffraction peak (Cu Ka radiation) selected from 8.47, 8.81, 12.75,13.17, 14.92,15.63,17.01 23.73, and 24.07 ± 0.20;

    ^Claims

    1. A crystalline ansolvate of Compound 1:

    2. The crystalline ansolvate of claim 1, which is substantially free of a solvated polymorph of Compound 1.

    3. A composition comprising the crystalline ansolvate of claim 1.

    4. The crystalline ansolvate of claim 1, wherein the crystalline ansolvate comprises one or more of Form I, Form II or Material N, wherein

    Form I is characterized by at least one X-ray powder diffraction peak (Cu Ka radiation) selected from 12.82°, 15.74°, 16.03°, 16.63°, 17.60°, 25.14°, 25.82° and 26.44° (each ±0.2 °20);

    Form II is characterized by at least one X-ray powder diffraction peak (Cu Ka radiation) selected from 13.37°, 14.37°, 19.95° and 23.92°20 (each ±0.2 °20); and Material N is characterized by at least one X-ray powder diffraction peak (Cu Ka radiation) selected from 11.65°, 11.85°, 12.08°, 16.70°, 19.65° and 23.48 °20 (each ±0.2 °20).

    5. The crystalline ansolvate of claim 4, wherein the crystalline ansolvate comprises Form IL

    6. The crystalline ansolvate of claim 4, wherein the crystalline ansolvate comprises Material N.

    7. A method of preparing a crystalline ansolvate of claim 1 comprising slurrying a sait of Compound 1 in a solvent and disproportionating the Compound 1 sait to form the crystalline ansolave.

    8. Use of a crystalline ansolvate of claim 1 for the manufacture of a pharmaceutical composition, for increasing oxygen affinity of hemoglobin S.

    Use of a composition of claim 3 for the manufacture of a pharmaceutical composition, for increasing oxygen affinity of hemoglobin S.

    10. Use of a crystalline ansolvate of claim 1 for the manufacture of a pharmaceutical composition, for treatment of oxygen deficiency associated with sickle cell anémia..

    11. Use of a composition of claim 3 for the manufacture of a pharmaceutical composition, for treatment of oxygen deficiency associated with sickle cell anémia.

    12. A crystalline solvaté of Compound 1:

    13. The crystalline solvaté of claim 12, which is substantially free of an ansolvated polymorph of Compound 1.

    14. A composition comprising the crystalline solvaté of claim 12 or claim 13.

    15. The crystalline solvaté of claim 12, comprising one or more of Material E, Material F, Material G, Material H, Material J, Material K, Material L, Material M, Material O or Material P.

    16. The crystalline solvaté of claim 15, wherein:

    Material E is characterized by at least one, two, or three X-ray powder diffraction peak (Cu Ka radiation) selected from 8.69, 11.73, 12.10, 15.26,16.11,17.45, 22.39, 22.55 and 23.70 ± 0.20;

    Material F is characterized by at least one, two, or three X-ray powder diffraction peak (Cu Ka radiation) selected from 8.47, 8.81, 12.75, 13.17,14.92, 15.63, 17.01 23.73, and 24.07 ±0.20;

    Material G is characterized by at least one, two, or three X-ray powder diffraction peak (Cu Ka radiation) selected from 8.47, 11.45, 12.62, 14.66, 15.69, 17.01, 18.47, 20.32, 22.61, 23.08, 23.43 and 23.70 ± 0.20;

    Material H is characterized by at least one, two, or three X-ray powder diffraction peak (Cu Ka radiation) selected from 8.61, 11.67, 15.33,16.28, 17.28, 22.58, 23.51 and 25.77 ±0.20;

    Material J is characterized by at least one, two, or three X-ray powder diffraction peak (Cu Ka radiation) selected from 8.52, 8.88, 12.79, 15.04, 15.61, 17.11, 22.81, 23.87, 24.17, 24.62 and 26.44 ± 0.20;

    Material K is characterized by at least one, two, or three X-ray powder diffraction peak (Cu Ka radiation) selected from 8.52; 8.83, 11.35, 15.04, 15.74, 17.11, 23.46, 23.58, 24.08 and 25.99 ± 0.20;

    Material L is characterized by at least one, two, or three X-ray powder diffraction peak (Cu Ka radiation) selected from 8.61, 8.78, 11.67, 14.94, 15.28, 16.14, 17.30, 22.75, 23.71 and 26.05 ± 0.20; and

    Material M is characterized by at least one, two, or three X-ray powder diffraction peak (Cu Ka radiation) selected from 7.74, 10.05, 12.82, 15.33, 16.80, 20.82, 21.14, 25.80 and 26.97 ± 0.20.
17. 17. A method of preparing a crystalline solvaté of claim 12 comprising contacting a free base ansolvate of Compound 1 with a solvent.
18. 18. Use of a crystalline solvaté of claim 12 for the manufacture of a pharmaceutical composition, for increasing oxygen affinity of hemoglobin S.
19. 19. Use of a crystalline solvaté of claim 12 for the manufacture of a pharmaceutical composition, for treatment of oxygen deficiency associated with sickle cell anémia.