Crystalline solid forms of 6-carboxy-2-(3,5-dichlorophenyl)-benzoxazole

Abstract

To provide a solid form of 6-carboxy-2-(3,5-dichlorophenyl)-benzoxazole, a pharmaceutical composition, and a method for treating transthyretin amyloid diseases.SOLUTION: A crystalline form of 6-carboxy-2-(3,5-dichlorophenyl)-benzoxazole is provided, wherein the crystalline form has a solid state NMR spectrum comprising 13C chemical shifts (ppm) at 120.8±0.2 and 127.7±0.2.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to the crystalline forms of 6-carboxy-2-(3,5-dichlorophenyl)-benzoxazole and methods for its preparation and use.

[0002] The synthetic route of 6-carboxy-2-(3,5-dichlorophenyl)-benzoxazole (hereinafter, "the compound of formula I") is described in U.S. Patent No. 7,214,695, and the solid forms of the meglumine salt of the compound of formula I are described in U.S. Patent Application No. 14 / 345,111 (the U.S. national stage of International Application No. PCT / IB2012 / 054748), all of which are hereby incorporated by reference in their entirety for all purposes. The compound of formula I has the structure shown below.

[0003]

Chemical Formula

[0004] The compound of formula I stabilizes the protein transthyretin (TTR) whose dissociation is involved in TTR amyloidosis (i.e., the compound of formula I prevents the dissociation of native TTR tetramers into monomers, thereby inhibiting TTR amyloid fibril formation) and has been developed for use in the treatment of transthyretin amyloid diseases.

Prior Art Documents

Patent Documents

[0005]

Patent Document 1

Patent Document 2

Patent Document 3

Patent Document 4

Patent Document 5

[0006] [Non-Patent Document 1] United States Pharmacopeia, USP37, NF32, S1<941>, from 503, 5 / 1 / 2014 [Non-Patent Document 2] http: / / www.ccdc.cam.ac.uk / mercury / [Non-Patent Document 3] Remington: The Science & Practice of Pharmacy, 19th Edition, Williams & Williams, (1995) [Non-Patent Document 4] "Physician’s Desk Reference", 52nd Edition, Medical Economics, Montvale, NJ (1998) [Non-Patent Document 5] "Handbook of Pharmaceutical Excipients", 3rd Edition, edited by A.H. Kibbe, Pharmaceutical Press, 2000 [Non-Patent Document 6] Colon, W.; Kelly, J.W., Biochemistry, 1992, 31, 8654 - 60 [Non-Patent Document 7] Kelly, J.W., Curr. Opin. Struct. Biol., 1996, 6, 11 - 7 [Non-Patent Document 8] Liu, K. et al., Nat. Struct. Biol., 2000, 7, 754 - 7 [Non-Patent Document 9] Westermark, P. et al., Proc. Natl. Acad. Sci. U.S.A., 1990, 87, 2843 - 5

Non-Patent Document 10

Non-Patent Document 11

Non-Patent Document 12

Non-Patent Document 13

Summary of the Invention

Problems to be Solved by the Invention

[0007] Solid forms are of interest to the pharmaceutical industry, particularly those involved in the development of appropriate dosage forms. If the solid form is not consistently maintained during clinical or stability studies, the exact dosage form used or studied may not be comparable from lot to lot. Also, when using a compound in clinical studies or commercial products, there is a risk of undesirable toxic effects due to the impurities present, so it is desirable to have a process for producing the compound with the selected solid form in high purity. Certain solid forms can also exhibit enhanced stability or can be more easily manufactured in high purity and in large quantities, and thus are more suitable for incorporation into pharmaceutical formulations. Certain solid forms may exhibit other favorable physical properties such as lack of hygroscopic tendency, filterability, improved solubility, and enhanced dissolution rate, which are due to various lattice energies.

[0008] The discussion of the background of the invention in this specification is included to explain the context of the invention. This is not an admission that any of the materials mentioned are publicly available, known, or part of common general knowledge in any country at the time of the priority date of any of the claims.

Means for Solving the Problems

[0009] This specification discloses solid forms of the compounds of Formula I, and each solid form can be uniquely identified alone or in combination by several different analytical parameters, including but not limited to powder X-ray diffraction pattern peaks or combinations of two or more peaks; solid-state NMR 13C chemical shifts or combinations of two or more chemical shifts; and Raman shift peaks or combinations of two or more Raman shift peaks.

[0010] Based on the disclosure provided herein, those skilled in the art will understand that the first crystalline form of the compound of Formula I (referred to herein as "Form 1") can be uniquely identified by several different spectral peaks or patterns in various combinations. The following are exemplary combinations of characteristic peak values that can be used for the identification of Form 1, and these exemplary combinations should not be construed as limiting any other combinations of peak values disclosed herein.

[0011] One aspect of the present invention provides a form which is Form 1 and has a powder X-ray diffraction pattern comprising peaks at diffraction angles (2θ) of 15.4 ± 0.2 and 20.2 ± 0.2. In another embodiment, Form 1 has a powder X-ray diffraction pattern comprising peaks at diffraction angles (2θ) of 15.4 ± 0.2, 20.2 ± 0.2, and 28.6 ± 0.2. In another embodiment, Form 1 has a powder X-ray diffraction pattern comprising peaks at diffraction angles (2θ) of 15.4 ± 0.2, 20.2 ± 0.2, 28.6 ± 0.2, and 29.0 ± 0.2. In another embodiment, Form 1 has a powder X-ray diffraction pattern comprising peaks at diffraction angles (2θ) of 15.4 ± 0.2, 20.2 ± 0.2, 23.5 ± 0.2, 28.6 ± 0.2, and 29.0 ± 0.2.

[0012] One aspect of the present invention provides a form in Form 1 having a powder X-ray diffraction pattern including peaks at diffraction angles (2θ) of 16.5 ± 0.2, 26.7 ± 0.2, and 28.6 ± 0.2. In another embodiment, Form 1 has a powder X-ray diffraction pattern including peaks at diffraction angles (2θ) of 16.5 ± 0.2, 26.7 ± 0.2, 28.6 ± 0.2, and 29.0 ± 0.2. In another embodiment, Form 1 has a powder X-ray diffraction pattern including peaks at diffraction angles (2θ) of 15.4 ± 0.2, 16.5 ± 0.2, 26.7 ± 0.2, 28.6 ± 0.2, and 29.0 ± 0.2.

[0013] Another aspect of the present invention provides a form in Form 1 having a powder X-ray diffraction pattern including peaks at diffraction angles (2θ) that are essentially the same as those shown in FIG. 1.

[0014] Another aspect of the present invention provides a form in Form 1 having a powder X-ray diffraction pattern including peaks at diffraction angles (2θ) that are essentially the same as those shown in FIG. 21.

[0015] Another aspect of the present invention provides a form in Form 1 having a Raman spectrum including Raman shift peaks (cm−1) at 287 ± 2, 869 ± 2, and 1292 ± 2. In another embodiment, Form 1 has a Raman spectrum including Raman shift peaks (cm−1) at 213 ± 2, 287 ± 2, 869 ± 2, and 1292 ± 2.

[0016] Another aspect of the present invention provides a form in Form 1 having a Raman spectrum including Raman shift peaks (cm−1) at 994 ± 2, 1273 ± 2, 1292 ± 2, and 1615 ± 2. In another embodiment, Form 1 has a Raman spectrum including Raman shift peaks (cm−1) at 213 ± 2, 994 ± 2, 1273 ± 2, 1292 ± 2, and 1615 ± 2.

[0017] Another aspect of the present invention provides a form in Form 1 having a Raman spectrum including Raman shift peaks (cm−1) at a position that is essentially the same as that shown in FIG. 5.

[0018] Another aspect of the present invention provides a form, Form 1, having a solid NMR spectrum containing 13C chemical shifts (ppm) at 120.8 ± 0.2, 127.7 ± 0.2, and 139.6 ± 0.2. In another embodiment, Form 1 has a solid NMR spectrum containing 13C chemical shifts (ppm) at 127.7 ± 0.2 and 139.6 ± 0.2. In another embodiment, Form 1 has a solid NMR spectrum containing 13C chemical shifts (ppm) at 120.8 ± 0.2 and 139.6 ± 0.2. In another embodiment, Form 1 has a solid NMR spectrum containing 13C chemical shifts (ppm) at 120.8 ± 0.2 and 127.7 ± 0.2.

[0019] Another aspect of the present invention provides a form, Form 1, having a solid NMR spectrum containing 13C chemical shifts (ppm) at 120.8 ± 0.2, 127.7 ± 0.2, and 144.7 ± 0.2. In another embodiment, Form 1 has a solid NMR spectrum containing 13C chemical shifts (ppm) at 127.7 ± 0.2 and 144.7 ± 0.2. In another embodiment, Form 1 has a solid NMR spectrum containing 13C chemical shifts (ppm) at 120.8 ± 0.2 and 144.7 ± 0.2. In another embodiment, Form 1 has a solid NMR spectrum containing 13C chemical shifts (ppm) at 120.8 ± 0.2 and 127.7 ± 0.2.

[0020] Another aspect of the present invention provides a form, Form 1, having a solid NMR spectrum containing 13C chemical shifts (ppm) at essentially the same positions as shown in FIG. 9.

[0021] Another aspect of the present invention provides a form, Form 1, having a powder X-ray diffraction pattern containing peaks at diffraction angles (2θ) of (i) 15.4 ± 0.2, 20.2 ± 0.2, and 28.6 ± 0.2, and a Raman spectrum containing Raman shift peaks (cm−1) at (ii) 287 ± 2, 869 ± 2, and 1292 ± 2.

[0022] Another aspect of the present invention is Form 1, which provides a form having (i) a powder X-ray diffraction pattern including a peak at a diffraction angle (2θ) of 28.6 ± 0.2, and (ii) a Raman spectrum including Raman shift peaks (cm-1) at 287 ± 2, 869 ± 2, and 1292 ± 2.

[0023] Another aspect of the present invention is Form 1, which provides a form having (i) a powder X-ray diffraction pattern including peaks at diffraction angles (2θ) of 15.4 ± 0.2, 20.2 ± 0.2, and 28.6 ± 0.2, and (ii) a solid NMR spectrum including 13C chemical shifts (ppm) at 120.8 ± 0.2 and 139.6 ± 0.2.

[0024] Another aspect of the present invention is Form 1, which provides a form having (i) a powder X-ray diffraction pattern including a peak at a diffraction angle (2θ) of 28.6 ± 0.2, and (ii) a solid NMR spectrum including 13C chemical shifts (ppm) at 120.8 ± 0.2 and 139.6 ± 0.2.

[0025] Another aspect of the present invention is Form 1, which provides a form having (i) a powder X-ray diffraction pattern including peaks at diffraction angles (2θ) of 26.7 ± 0.2 and 28.6 ± 0.2, and (ii) a solid NMR spectrum including a 13C chemical shift (ppm) at 127.7 ± 0.2.

[0026] Another aspect of the present invention is Form 1, which provides a form having (i) a Raman spectrum including Raman shift peaks (cm-1) at 287 ± 2, 869 ± 2, and 1292 ± 2, and (ii) a solid NMR spectrum including 13C chemical shifts (ppm) at 120.8 ± 0.2 and 139.6 ± 0.2.

[0027] Another aspect of the present invention is Form 1, which provides a form having (i) a Raman spectrum including Raman shift peaks (cm-1) at 994 ± 2, 1273 ± 2, and 1292 ± 2, and (ii) a solid NMR spectrum including 13C chemical shifts (ppm) at 120.8 ± 0.2 and 127.7 ± 0.2.

[0028] Another aspect of the present invention is Form 1, which provides a form having (i) a Raman spectrum including Raman shift peaks (cm-1) at 1292 ± 2 and 1615 ± 2, and (ii) a solid NMR spectrum including a 13C chemical shift (ppm) at 127.7 ± 0.2.

[0029] In certain embodiments, the present invention relates to Form 1 which is non-hygroscopic and anhydrous.

[0030] In certain embodiments, the present invention relates to Form 1 which comprises a plurality of acicular crystals of the compound of Formula I.

[0031] In a further aspect, the present invention contemplates that Form 1 can be present in the presence of any of the other solid forms thereof (e.g., Form 2, 4, and 6) or mixtures thereof. Thus, in one embodiment, the present invention provides Form 1 present in a solid form comprising less than 95 wt%, less than 90 wt%, less than 80 wt%, less than 70 wt%, less than 60 wt%, less than 50 wt%, less than 40 wt%, less than 30 wt%, less than 20 wt%, less than 10 wt%, less than 5 wt%, less than 3 wt%, or less than 1 wt% of any other physical form of the compound of Formula I. For example, in one embodiment, it is a solid form of the compound of Formula I comprising Form 1 having any one of the above powder X-ray diffraction pattern, Raman spectrum, IR spectrum, and / or NMR spectrum, and the solid form comprises less than 95 wt%, less than 90 wt%, less than 80 wt%, less than 70 wt%, less than 60 wt%, less than 50 wt%, less than 40 wt%, less than 30 wt%, less than 20 wt%, less than 10 wt%, less than 5 wt%, less than 3 wt%, or less than 1 wt% of any other physical form of the compound of Formula I.

[0032] In certain embodiments, the present invention relates to Form 1 which is in a substantially pure crystalline form.

[0033] Furthermore, based on the disclosure provided herein, one of ordinary skill in the art will understand that the second crystalline form of the compound of Formula I (referred to herein as "Form 4") can be uniquely identified by several different spectral peaks or patterns in various combinations. The following are exemplary combinations of characteristic peak values that can be used to identify Form 4, and these exemplary combinations should in no way be construed as limiting other combinations of peak values disclosed herein.

[0034] One aspect of the present invention provides a form that is Form 4 and has a powder X-ray diffraction pattern including peaks at diffraction angles (2θ) of 15.9 ± 0.2 and 16.9 ± 0.2. In another embodiment, Form 4 has a powder X-ray diffraction pattern including peaks at diffraction angles (2θ) of 15.9 ± 0.2, 16.9 ± 0.2, and 18.0 ± 0.2. In another embodiment, Form 4 has a powder X-ray diffraction pattern including peaks at diffraction angles (2θ) of 16.9 ± 0.2, 24.1 ± 0.2, and 27.3 ± 0.2. In another embodiment, Form 4 has a powder X-ray diffraction pattern including peaks at diffraction angles (2θ) of 15.9 ± 0.2, 16.9 ± 0.2, 18.0 ± 0.2, and 27.3 ± 0.2.

[0035] Another aspect of the present invention provides a crystalline form that is Form 4 and has a powder X-ray diffraction pattern including peaks at diffraction angles (2θ) that are essentially the same as those shown in Figure 3.

[0036] Another aspect of the present invention provides a form that is Form 4 and has a Raman spectrum including Raman shift peaks (cm-1) at 266 ± 2, 283 ± 2, and 1297 ± 2. In another embodiment, Form 4 has a Raman spectrum including Raman shift peaks (cm-1) at 201 ± 2, 266 ± 2, 283 ± 2, and 1297 ± 2. In another embodiment, Form 4 has a Raman spectrum including Raman shift peaks (cm-1) at 283 ± 2, 994 ± 2, 1273 ± 2, and 1547 ± 2.

[0037] Another aspect of the present invention is Form 4, which provides a form having a Raman spectrum containing Raman shift peaks (cm-1) at essentially the same positions as shown in FIG. 7.

[0038] Another aspect of the present invention is Form 4, which provides a form having a solid NMR spectrum containing 13C chemical shifts (ppm) at 122.1 ± 0.2, 130.7 ± 0.2, and 140.1 ± 0.2. In another embodiment, Form 4 has a solid NMR spectrum containing 13C chemical shifts (ppm) at 122.1 ± 0.2, 124.4 ± 0.2, and 130.7 ± 0.2. In another embodiment, Form 4 has a solid NMR spectrum containing 13C chemical shifts (ppm) at 130.7 ± 0.2 and 140.1 ± 0.2. In another embodiment, Form 4 has a solid NMR spectrum containing 13C chemical shifts (ppm) at 122.1 ± 0.2 and 140.1 ± 0.2. In another embodiment, Form 4 has a solid NMR spectrum containing 13C chemical shifts (ppm) at 122.1 ± 0.2 and 130.7 ± 0.2. In another embodiment, Form 4 has a solid NMR spectrum containing 13C chemical shifts (ppm) at 124.4 ± 0.2 and 130.7 ± 0.2.

[0039] Another aspect of the present invention is Form 4, which provides a form having a solid NMR spectrum containing 13C chemical shifts (ppm) at essentially the same positions as shown in FIG. 11.

[0040] Another aspect of the present invention is Form 4, which provides a form having (i) a powder X-ray diffraction pattern containing peaks at diffraction angles (2θ) of 15.9 ± 0.2 and 16.9 ± 0.2, and (ii) a Raman spectrum containing Raman shift peaks (cm-1) at 266 ± 2, 283 ± 2, and 1297 ± 2. Another aspect of the present invention is Form 4, which provides a form having (i) a powder X-ray diffraction pattern containing peaks at diffraction angles (2θ) of 15.9 ± 0.2 and 16.9 ± 0.2, and (ii) a solid NMR spectrum containing 13C chemical shifts (ppm) at 122.1 ± 0.2, 130.7 ± 0.2, and 140.1 ± 0.2.

[0041] Another aspect of the present invention is Form 4, which provides a form having a Raman spectrum including Raman shift peaks (cm−1) at 266±2, 283±2, and 1297±2, and a solid-state NMR spectrum including 13C chemical shifts (ppm) at 122.1±0.2, 130.7±0.2, and 140.1±0.2.

[0042] In certain embodiments, the present invention relates to Form 4 which is non-hygroscopic and anhydrous.

[0043] In certain embodiments, the present invention relates to Form 4 which comprises a plurality of acicular crystals of the compound of formula I.

[0044] In a further aspect, the present invention contemplates that Form 4 can be present in the presence of any of the other solid forms thereof (e.g., Forms 1, 2, and 6) or mixtures thereof. Thus, in one embodiment, the present invention provides Form 4 present in a solid form comprising less than 95 wt%, less than 90 wt%, less than 80 wt%, less than 70 wt%, less than 60 wt%, less than 50 wt%, less than 40 wt%, less than 30 wt%, less than 20 wt%, less than 10 wt%, less than 5 wt%, less than 3 wt%, or less than 1 wt% of any other physical form of the compound of formula I. For example, in one embodiment, it is a solid form of the compound of formula I comprising Form 4 having any one of the above-described powder X-ray diffraction pattern, Raman spectrum, IR spectrum, and / or NMR spectrum, and the solid form comprises less than 95 wt%, less than 90 wt%, less than 80 wt%, less than 70 wt%, less than 60 wt%, less than 50 wt%, less than 40 wt%, less than 30 wt%, less than 20 wt%, less than 10 wt%, less than 5 wt%, less than 3 wt%, or less than 1 wt% of any other physical form of the compound of formula I.

[0045] In certain embodiments, the present invention relates to Form 4 which is in a substantially pure crystalline form.

[0046] Furthermore, based on the disclosure provided herein, one of ordinary skill in the art will understand that the third crystalline form of the compound of Formula I (referred to herein as "Form 6") can be uniquely identified by several different spectral peaks or patterns in various combinations. The following are exemplary combinations of characteristic peak values that can be used to identify Form 6, and these exemplary combinations should in no way be construed as limiting other combinations of peak values disclosed herein.

[0047] One aspect of the present invention provides a form that is Form 6 and has a powder X-ray diffraction pattern including peaks at diffraction angles (2θ) of 23.8 ± 0.2 and 27.5 ± 0.2. In another embodiment, Form 6 has a powder X-ray diffraction pattern including peaks at diffraction angles (2θ) of 13.6 ± 0.2, 23.8 ± 0.2, and 27.5 ± 0.2. In another embodiment, Form 6 has a powder X-ray diffraction pattern including peaks at diffraction angles (2θ) of 13.6 ± 0.2, 23.5 ± 0.2, 23.8 ± 0.2, and 27.5 ± 0.2.

[0048] Another aspect of the present invention provides a crystalline form that is Form 6 and has a powder X-ray diffraction pattern including peaks at diffraction angles (2θ) that are essentially the same as those shown in FIG. 14.

[0049] Another aspect of the present invention provides a form that is Form 6 and has a Raman spectrum including Raman shift peaks (cm-1) at 223 ± 2, 1274 ± 2, and 1434 ± 2. In another embodiment, Form 6 has a Raman spectrum including Raman shift peaks (cm-1) at 223 ± 2, 1274 ± 2, 1434 ± 2, and 1547 ± 2.

[0050] Another aspect of the present invention provides a form that is Form 6 and has a Raman spectrum including Raman shift peaks (cm-1) at a position that is essentially the same as that shown in FIG. 16.

[0051] Another aspect of the present invention is Form 6, which provides a form having a solid-state NMR spectrum including 13C chemical shifts (ppm) at 109.7 ± 0.2, 126.4 ± 0.2, and 131.5 ± 0.2. In another embodiment, Form 6 has a solid-state NMR spectrum including 13C chemical shifts (ppm) at 109.7 ± 0.2 and 126.4 ± 0.2. In another embodiment, Form 6 has a solid-state NMR spectrum including 13C chemical shifts (ppm) at 126.4 ± 0.2 and 131.5 ± 0.2. In another embodiment, Form 6 has a solid-state NMR spectrum including 13C chemical shifts (ppm) at 109.7 ± 0.2 and 131.5 ± 0.2.

[0052] Another aspect of the present invention is Form 6, which provides a form having a solid-state NMR spectrum including 13C chemical shifts (ppm) at essentially the same positions as shown in FIG. 18.

[0053] Another aspect of the present invention is Form 6, which provides a form having (i) a powder X-ray diffraction pattern including peaks at diffraction angles (2θ) of 23.8 ± 0.2 and 27.5 ± 0.2, and (ii) a Raman spectrum including Raman shift peaks (cm−1) at 223 ± 2, 1274 ± 2, and 1547 ± 2.

[0054] Another aspect of the present invention is Form 6, which provides a form having (i) a powder X-ray diffraction pattern including peaks at diffraction angles (2θ) of 23.8 ± 0.2 and 27.5 ± 0.2, and (ii) a solid-state NMR spectrum including 13C chemical shifts (ppm) at 109.7 ± 0.2, 126.4 ± 0.2, and 131.5 ± 0.2.

[0055] Another aspect of the present invention is Form 6, which provides a form having (i) a Raman spectrum including Raman shift peaks (cm−1) at 223 ± 2, 1274 ± 2, and 1547 ± 2, and (ii) a solid-state NMR spectrum including 13C chemical shifts (ppm) at 109.7 ± 0.2, 126.4 ± 0.2, and 131.5 ± 0.2.

[0056] In certain embodiments, the present invention relates to Form 6 which is non-hygroscopic and anhydrous.

[0057] In a further aspect, the present invention contemplates that Form 6 can be present in the presence of any of its other solid forms (e.g., Forms 1, 2, and 4) or mixtures thereof. Thus, in one embodiment, the present invention provides Form 6 present in a solid form comprising less than 95 wt%, less than 90 wt%, less than 80 wt%, less than 70 wt%, less than 60 wt%, less than 50 wt%, less than 40 wt%, less than 30 wt%, less than 20 wt%, less than 10 wt%, less than 5 wt%, less than 3 wt%, or less than 1 wt% of any other physical form of the compound of Formula I. For example, in one embodiment, it is a solid form of the compound of Formula I comprising Form 6 having any one of the above-mentioned powder X-ray diffraction pattern, Raman spectrum, IR spectrum, and / or NMR spectrum, and the solid form comprises less than 95 wt%, less than 90 wt%, less than 80 wt%, less than 70 wt%, less than 60 wt%, less than 50 wt%, less than 40 wt%, less than 30 wt%, less than 20 wt%, less than 10 wt%, less than 5 wt%, less than 3 wt%, or less than 1 wt% of any other physical form of the compound of Formula I.

[0058] In certain embodiments, the present invention relates to Form 6 which is in a substantially pure crystalline form.

[0059] A further aspect of the present invention provides a pharmaceutical composition comprising Form 1, Form 2, Form 4, or Form 6 described herein. In a further aspect, the present invention provides an oral dosage form comprising any one of Form 1, Form 2, Form 4, or Form 6, or the pharmaceutical composition described herein. For example, in one embodiment, the oral dosage form is a tablet, pill, or capsule. For example, in one embodiment, the oral dosage form is a tablet or capsule.

[0060] In one embodiment, the present invention provides a tablet comprising any one of Form 1, Form 2, Form 4, or Form 6 described herein, or a pharmaceutical composition. For example, in one embodiment, the tablet comprises from about 1 to about 100 mg of Form 1, 2, 4, or 6. Further, for example, the tablet comprises about 10 mg of Form 1, 2, 4, or 6. Further, for example, the tablet comprises about 20 mg of Form 1, 2, 4, or 6. Further, for example, the tablet comprises about 30 mg of Form 1, 2, 4, or 6. Further, for example, the tablet comprises about 40 mg of Form 1, 2, 4, or 6. Further, for example, the tablet comprises about 50 mg of Form 1, 2, 4, or 6. Further, for example, the tablet comprises about 60 mg of Form 1, 2, 4, or 6. Further, for example, the tablet comprises about 70 mg of Form 1, 2, 4, or 6. Further, for example, the tablet comprises about 80 mg of Form 1, 2, 4, or 6. Further, for example, the tablet comprises about 90 mg of Form 1, 2, 4, or 6. Further, for example, the tablet comprises about 100 mg of Form 1, 2, 4, or 6.

[0061] In one embodiment, the present invention provides a soft gelatin capsule comprising any one of Form 1, Form 2, Form 4, Form 6, or the pharmaceutical composition described herein. For example, in one embodiment, the soft gelatin capsule comprises about 1 to about 100 mg of Form 1, 2, 4, or 6. Further, for example, the soft gelatin capsule comprises about 10 mg of Form 1, 2, 4, or 6. Further, for example, the soft gelatin capsule comprises about 20 mg of Form 1, 2, 4, or 6. Further, for example, the soft gelatin capsule comprises about 30 mg of Form 1, 2, 4, or 6. Further, for example, the soft gelatin capsule comprises about 40 mg of Form 1, 2, 4, or 6. Further, for example, the soft gelatin capsule comprises about 50 mg of Form 1, 2, 4, or 6. Further, for example, the soft gelatin capsule comprises about 60 mg of Form 1, 2, 4, or 6. Further, for example, the soft gelatin capsule comprises about 70 mg of Form 1, 2, 4, or 6. Further, for example, the soft gelatin capsule comprises about 80 mg of Form 1, 2, 4, or 6. Further, for example, the soft gelatin capsule comprises about 90 mg of Form 1, 2, 4, or 6. Further, for example, the soft gelatin capsule comprises about 100 mg of Form 1, 2, 4, or 6.

[0062] A further aspect of the present invention provides a method for preparing Form 1 as described in Example 1. A further aspect of the present invention provides a method for preparing Form 4, the method comprising heating Form 1 as described in Example 2. A further aspect of the present invention provides a method for preparing Form 2, the method comprising dissolving Form 1 in THF and evaporating the resulting solution as described in Example 3. A further aspect of the present invention provides a method for preparing Form 6, the method comprising heating Form 1 as described in Example 4.

[0063] A further aspect of the present invention is a method for treating transthyretin amyloid diseases such as senile systemic amyloidosis (SSA), familial amyloid polyneuropathy (FAP), and familial amyloid cardiomyopathy (FAC) in mammals, the method comprising administering to the mammal a therapeutically effective amount of any one of Form 1, Form 2, Form 4, Form 6, or the pharmaceutical composition described herein.

Brief Description of the Drawings

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Embodiments for Carrying Out the Invention

[0065] Based on the chemical structure, it is not possible to predict with a certain degree of certainty whether a compound will crystallize, under what conditions it will crystallize, how many crystalline solid forms of the compound may exist, or the solid structure of any of those forms. An important characteristic of any crystalline drug is the polymorphic behavior of such materials. Generally, the crystalline form of a drug is preferred over its amorphous form. This is partly due to their excellent stability. For example, in many situations, during storage, an amorphous drug converts to the crystalline form of the drug. Since the amorphous and crystalline forms of a drug typically have different physical and chemical properties, such interconversion may be undesirable for safety reasons in the use of pharmaceuticals. The different physical properties exhibited by different solid forms of a pharmaceutical compound can affect important pharmaceutical parameters, such as storage, stability, compressibility, density (which are important in the manufacture of formulations and products), and dissolution rate (which is important in the determination of bioavailability). Differences in stability can result from changes in chemical reactivity (e.g., hydrolysis or oxidation is different, such that a dosage form containing a certain polymorph may discolor more rapidly than a dosage form containing a different polymorph), mechanical changes (e.g., if the kinetically preferred crystalline form converts to the thermodynamically more stable crystalline form, the tablet may disintegrate during storage), or both (e.g., a tablet of one polymorph may be more susceptible to degradation at high humidity). In extreme situations, differences in solubility between polymorphs can lead to a shift to a crystalline form that lacks efficacy and / or exhibits toxicity. Furthermore, the physical properties of the crystalline form can also be important in the processing of pharmaceuticals. For example, a particular crystalline form may be more likely to form solvates, or it may be more difficult to filter and wash to remove impurities than other crystalline forms (i.e., the particle shape and size distribution may differ between one crystalline form and another).

[0066] There is no single ideal physical form of a drug because different physical forms offer different advantages. Searching for the most stable form and other such forms requires a great deal of effort, and the results are unpredictable. Therefore, it is important to seek diverse and unique drug forms, such as salts, polymorphs, and amorphous forms, that can be used in various formulations. Selecting a drug form for a specific formulation or therapeutic use requires consideration of various properties, and the best form for a particular use can have one specific important favorable property, while other properties may be acceptable or marginally acceptable.

[0067] The success of drug development requires meeting certain general requirements for it to be a therapeutically effective treatment for patients. These requirements are classified into two categories: (1) requirements for successfully manufacturing the dosage form, and (2) requirements for successfully delivering the drug and achieving its pharmacokinetics after administering the drug formulation to the patient.

[0068] Different crystalline solid forms of the same compound often have different solid properties, such as melting point, solubility, dissolution rate, hygroscopicity, powder flowability, mechanical properties, chemical stability, and physical stability. These solid properties can provide advantages in filtration, drying, and the operation of dosage form manufacturing units. Therefore, once different crystalline solid forms of the same compound are identified, the optimal crystalline solid form under any given set of processing and manufacturing conditions, as well as the different solid properties of each crystalline solid form, can be determined.

[0069] Molecular polymorphs can be obtained by several methods known in the art. Such methods include, but are not limited to, melt recrystallization, melt cooling, solvent recrystallization, desolvation, rapid evaporation, rapid cooling, slow cooling, vapor diffusion, and sublimation. Polymorphs can be detected, identified, classified, and characterized using well-known techniques such as, but not limited to, differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), X-ray powder diffraction (PXRD), single crystal X-ray diffraction, solid state nuclear magnetic resonance (NMR), infrared (IR) spectroscopy, Raman spectroscopy, and hot stage microscopy. With respect to drug development, it is important to provide a compound form (commonly known as the active pharmaceutical ingredient) that can be prepared and purified on a large scale with certainty, is stable upon storage, and does not decompose. Furthermore, the active pharmaceutical ingredient must be suitable for formulation into a dosage form selected according to the intended route of administration.

[0070] It has been found that the compounds of formula I can exist in distinct crystalline forms referred to herein as Form 1, Form 2, Form 4, and Form 6. These forms can be used in pharmaceutical products for the treatment of transthyretin amyloidosis. As described above, each form can have advantages over others with respect to properties such as bioavailability, stability, and manufacturability. In one aspect of the invention, crystalline forms of the compounds of formula I, namely Form 1, Form 2, Form 4, and Form 6, have been discovered, and these appear to be more suitable for large-scale preparation and handling than the amorphous forms. Processes for producing Forms 1, 2, 4, and 6 in high purity are described herein. Another object of the invention is to provide a process for preparing each solid form of the compounds of formula I substantially free of other solid forms. Further, an object of the invention is to provide a pharmaceutical formulation comprising the compounds of formula I in various solid forms as described above, and a method of treating transthyretin amyloidosis by administering such a pharmaceutical formulation.

[0071] Definitions As used herein, the term "treat" means, unless otherwise indicated, to restore, alleviate, inhibit the progression of, or prevent a disorder or condition to which such term applies, or one or more symptoms of such disorder or condition. As used herein, the term "treatment" refers to the act of "treating" as defined immediately above, unless otherwise indicated.

[0072] As used herein, transthyretin or TTR is a 55 kDa homotetramer characterized by 2,2,2 symmetry, having two identical funnel-shaped binding sites at the dimer-dimer interface, at which sites thyroid hormone (T4) can bind in plasma and CSF. TTR generally binds to less than 1 equivalent of holo-retinol binding protein. TTR is a 127-residue protein that tetramerizes under physiological conditions. TTR functions as the third transporter of thyroxine in serum and as the first carrier in cerebrospinal fluid. TTR also transports retinol by association with retinol binding protein. TTR forms amyloid at low pH.

[0073] As used herein, the term "substantially pure" in relation to a particular crystalline form means that the crystalline or amorphous form contains less than 10% by weight, preferably less than 5% by weight, preferably less than 3% by weight, preferably less than 1% by weight of any other physical form of that compound.

[0074] As used herein, the term "substantially identical" in relation to the peak positions of X-ray diffraction means that typical peak positions and intensity variations are taken into account. For example, those skilled in the art will understand that the peak position (2θ) will exhibit some variation, generally on the order of 0.1 to 0.2 degrees, even in the apparatus used to measure the diffraction. Further, those skilled in the art will understand that the relative peak intensity exhibits variations due to variations between apparatuses, as well as variations due to crystallinity, preferred orientation, the surface of the prepared sample, and other factors known to those skilled in the art, and this should be regarded simply as a qualitative measure. Similarly, as used herein, "substantially identical" in relation to solid-state NMR spectra and Raman spectra is intended to encompass the variations associated with these analytical techniques known to those skilled in the art. For example, the 13C chemical shift measured by solid-state NMR generally has a variation of up to 0.2 ppm for very sharp peaks and even greater variation for broad lines, while the Raman shift generally has a variation of about 2 cm-1.

[0075] The term "polymorph" refers to the various crystalline forms of the same compound, which includes, but is not limited to, hydrates (e.g., water of crystallization is present in the crystal structure) and solvates (e.g., bound solvents other than water) of the same compound, as well as other solid molecular forms.

[0076] The term "amorphous" refers to any solid material lacking three-dimensional order. In some cases, amorphous solids can be characterized by known techniques including X-ray powder diffraction (PXRD) crystallography, solid-state nuclear magnetic resonance (ssNMR) spectroscopy, differential scanning calorimetry (DSC), or some combination of these techniques.

[0077] The term "crystalline" refers to any solid material exhibiting three-dimensional regularity, which, in contrast to amorphous solid materials, gives rise to a distinct PXRD pattern with sharp, well-defined peaks.

[0078] The term "solvate" refers to a molecular complex that includes the active ingredient and one or more solvent molecules (e.g., ethanol) in stoichiometric or non-stoichiometric amounts. When the solvent is tightly bound to the drug, the resulting complex has a well-defined, humidity-independent stoichiometry. However, when the solvent is weakly bound, as in the case of channel solvates and hygroscopic compounds, the solvent content will depend on humidity and the state of dryness. In such cases, the complex will typically be non-stoichiometric.

[0079] The term "hydrate" refers to a solvate that includes the active ingredient and water in stoichiometric or non-stoichiometric amounts.

[0080] The term "powder X-ray diffraction pattern" or "PXRD pattern" refers to the diffractogram observed experimentally or parameters derived therefrom. The powder X-ray diffraction pattern is characterized by peak position (abscissa) and peak intensity (ordinate).

[0081] The term "2-theta value" or "2θ" refers to the peak position expressed in degrees, based on the experimental setup of the X-ray diffraction experiment, and is the common abscissa unit in the diffraction pattern. The experimental setup requires that when the incident beam forms an angle theta (θ) with a fixed lattice plane and the reflection is diffracted, the reflected beam is recorded at an angle 2-theta (2θ). It should be understood that references in this specification to specific 2θ values for a particular solid form are intended to mean the 2θ values (degrees) measured using the X-ray diffraction experimental conditions described in this specification.

[0082] Solid forms of the compound of formula I The solid forms of the compounds of formula I disclosed herein can be characterized by one or more of the following: powder X-ray diffraction patterns (i.e., X-ray diffraction peaks at various diffraction angles (2θ)), solid state nuclear magnetic resonance (NMR) spectral patterns, Raman spectral diagram patterns, infrared spectral patterns, water solubility, photostability under the high-intensity light conditions of the International Conference on Harmonization (ICH), and physical and chemical storage stability. For example, the solid forms of the compounds of formula I were characterized by the positions and relative intensities of the peaks in their powder X-ray diffraction patterns, respectively.

[0083] The powder X-ray diffraction patterns of the solid forms of the compounds of formula I were collected on a PANalytical X’Pert PRO MPD diffractometer using an incident beam of Cu radiation generated using an Optix long high-precision focus source. An elliptical graded multilayer mirror was used to focus the Cu Kα X-rays onto the detector through the specimen. Prior to analysis, a silicon specimen (NIST SRM 640d) was analyzed to confirm that the observed Si111 peak position was in agreement with the NIST-certified position. The specimen of the sample was sandwiched between 3-μm thick films and analyzed in transmission geometry. A beam stop, a short scatter-line removal extension, and a knife edge for scatter-line removal were used to minimize the background generated by the atmosphere. Solar slits for the incident and diffracted beams were used to minimize the spread from axial divergence. The diffraction patterns were collected using a scanning position-sensitive detector (X’Celerator) positioned 240 mm from the specimen, and Data Collector version 2.2b software. The data acquisition parameters were as shown in Table 1 below.

[0084] [Table 1]

[0085] More generally, to perform X-ray diffraction measurements with a transmission instrument such as the PANalytical system used for the measurements reported herein, a specimen of the sample was sandwiched between 3-μm thick films and analyzed in a transmission geometry. The incident X-ray beam was first directed at the sample at a small angle with respect to the plane of the holder, and then moved through an arc that continuously increased the angle between the incident beam and the plane of the holder. Measurement errors associated with such X-ray powder analysis arise from various factors, including (a) errors in sample preparation; (b) instrument errors; (c) calibration errors; (d) operator errors (including errors present in determining peak positions); and (e) properties of the material (e.g., errors in preferred orientation and transmissibility). Calibration errors and sample height errors generally shift all peaks in the same direction. These shifts can be identified from the X-ray diffractogram and removed by compensating for the shifts (applying systematic correction factors to all peak position values) or by recalibrating the instrument. Usually, this correction factor brings the measured peak positions into agreement with the predicted peak positions, which need only be within the range of ±0.2° 2θ.

[0086] Those skilled in the art will understand that peak positions (2θ) will generally exhibit a variation between instruments of ±0.2° 2θ. Thus, when peak positions (2θ) are reported, those skilled in the art will recognize that such numbers are intended to subsume such variation between instruments. Further, when the crystalline forms of the present invention are described as having powder X-ray diffraction peak positions that are essentially the same as those shown in a given drawing, the term "essentially the same" is also intended to subsume such variation between instruments in the diffraction peak positions. Further, those skilled in the art will understand that relative peak intensities exhibit variation due to variation between instruments as well as variation due to crystallinity, preferred orientation, the surface of the sample prepared, and other factors known to those skilled in the art, and that this should be regarded simply as a qualitative measure.

[0087] The identification of PXRD peaks was performed as follows. The PXRD patterns were analyzed for Forms 1 and 4, and the effects of preferred orientation and particle statistics were not evaluated. In most situations, peaks within the range up to about 30° 2θ were selected. Peaks having an intensity of 2% or more of the strongest peak were used for peak selection. The peak positions were rounded to the nearest 0.1° 2θ. The peak positions (° 2θ) along the X-axis were determined using TRIADS (trademark) v2.0 software. The TRIADS (trademark) algorithm is described in U.S. Patent No. 8,576,985, which is hereby incorporated by reference in its entirety. As described above, the variation in peak positions is shown within a range of ±0.2° 2θ based on the recommendations outlined in the USP's consideration of the variation in X-ray powder diffraction (see U.S. Pharmacopeia, USP37, NF32, S1 <941>, 503, 5 / 1 / 2014).

[0088] The solid forms of the compounds of Formula I can also be characterized by Raman spectroscopy. Raman spectra were collected using an NXR FT-Raman module connected to a Nexus 670 FT-IR spectrophotometer (Thermo Nicolet) equipped with an InGaAs detector. Sulfur and cyclohexane were used for wavelength verification. For analysis, the sample material was loaded into a pellet holder to prepare each sample. An Nd:YVO4 laser output of about 0.5 W (excitation wavelength 1064 nm) was used for irradiation of each sample. Each spectrum represents 256 simultaneous scans collected at a spectral resolution of 2 cm-1 at ambient temperature. The peak positions were picked at the maximum of the peaks. The relative intensity values were classified as strong (S), medium (M), and weak (W) using the following criteria: strong (1.00 - 0.75), medium (0.74 - 0.30), and weak (0.29 and below).

[0089] The solid forms of the compounds of Formula I can also be characterized using solid-state NMR spectroscopy. The 13C solid spectra for the solid forms of Formula 1 were collected as follows. Solid-state NMR (ssNMR) analysis was performed using a Bruker-Biospin CPMAS probe placed inside a Bruker-Biospin Avance III 500 MHz (1H frequency) NMR spectrometer at ambient temperature and pressure. The filled rotor was oriented at the magic angle and rotated at 15.0 kHz. The carbon ssNMR spectra were collected using a proton decoupling cross-polarization magic angle spinning (CPMAS) experiment at ambient temperature. During acquisition of the spectra, a phase-modulated proton decoupling field of 80 - 100 kHz was applied. The cross-polarization contact time was set to 2.0 milliseconds. The recycle delays were set to 180 seconds for Form 1, 50 seconds for Form 4, and 5 seconds for Form 6. The number of scans was adjusted to obtain an appropriate signal-to-noise ratio. The crystalline adamantane of the external standard was used, and its upfield resonance was set to 29.5 ppm (determined from neat tetramethylsilane), and its carbon spectrum was used as a reference. Automatic peak picking was performed using Bruker-BioSpin TopSpin version 3.1 software. Usually, a threshold of 5% relative intensity was used to preselect peaks. The output of the automatic peak picking was visually confirmed to ensure validity, and manual adjustment was made if necessary. Specific 13C solid NMR peak values are reported herein, but these peak values have a range due to differences in equipment, sample, and sample preparation. Since there is inherent variability in the peak values, this is conventional in the field of solid-state NMR. The typical variability for the X-axis values of 13C chemical shifts is about ±0.2 ppm for crystalline solids. The solid NMR peak heights reported herein are relative intensities. The solid NMR intensities may vary depending on the actual settings of the CPMAS experimental parameters and the thermal history of the sample.

[0090] One of ordinary skill in the art will recognize that the crystalline form of a given compound may exist in a substantially pure form of a single polymorph, but may also exist in a crystalline form that includes two or more different polymorphic or amorphous forms. When the solid form includes two or more polymorphs, the X-ray diffraction pattern will have peaks characteristic of each of the individual polymorphs of the invention. For example, a solid form that includes two polymorphs will have a powder X-ray diffraction pattern that is a superposition of the two X-ray diffraction patterns corresponding to the substantially pure solid forms. For example, a solid form of a compound of Formula I can contain a first and a second solid form, where the solid form contains at least 10% by weight of the first form. In a further example, the solid form contains at least 20% by weight of the first form. Yet further examples contain at least 30% by weight, at least 40% by weight, or at least 50% by weight of the first form. One of ordinary skill in the art will recognize that many such combinations of several individual forms in various amounts are possible.

[0091] Form 1 Form 1 is a crystalline, non-hygroscopic, anhydrous form of the compound of Formula I that can be produced as described in Example 1.

[0092] Form 1 was characterized by the PXRD pattern shown in Figure 1, measured on a PANalytical X’Pert PRO MPD using an incident beam of Cu radiation generated using an Optix long high-precision focal source. The PXRD pattern of Form 1, represented by degrees (2θ) and relative intensity with a relative intensity of ≥2.0%, is shown in Figure 2. The relative intensity can vary depending on the crystal size and form.

[0093] In this specification, Form 1 was taken as the neat substance in order to identify the characteristic peaks of Form 1 using appropriate analytical methods. From these analytical methods, peak values that are characteristics of Form 1 are provided that have a defined range within an acceptable variation. However, when Form 1 is mixed with any additional components, such as components utilized in a formulation, it is expected that the relative intensities of these characteristic peaks will change. Therefore, it will be understood by those skilled in the art of instrumental analysis that when Form 1 is mixed with or diluted by additional components within the scope of a pharmaceutical formulation, the analytical parameters of a particular method may require another optimization to enable the detection of these characteristic peaks. For example, as described in the following paragraphs, the PXRD method can be further optimized to enable the detection of characteristic Form 1 peaks when Form 1 is mixed with additional components. It will be understood by those skilled in the art of PXRD analysis that the peak values associated with the characteristic peaks of Form 1 will not be changed as a result of the optimization of this method.

[0094] Powder X-ray diffraction analysis of Form 1 was also performed using a Bruker AXS D8 ADVANCE diffractometer equipped with a Cu radiation source (K-α average). This system is equipped with a 2.5-axis Soller slit on the primary side. On the secondary side, a 2.5-axis Soller slit and an electric slit are utilized. The diffracted radiation was detected by a Lynx Eye XE detector. The X-ray tube voltage and amperage were set at 40 kV and 40 mA, respectively. Data were collected with a theta-theta goniometer up to 2 theta of 3.0 to 40.0 degrees at a Cu wavelength using a step size of 0.037 degrees and a step time of 10 seconds. The sample was prepared in a low-background holder and rotated during collection. The resulting powder pattern of Form 1 is shown in Figure 21.

[0095] Form 1 was also characterized by the pattern of the Raman spectrum shown in Figure 5, which was performed with an NXR FT-Raman module connected to a Nexus 670 FT-IR spectrometer (Thermo Nicolet) equipped with an InGaAs detector. The peaks of the Raman spectrum of Form 1 are shown in Figure 6.

[0096] Also, Form 1 was characterized by the pattern of the solid-state NMR spectrum shown in FIG. 9, which was performed with a Bruker-Biospin 4 mm CPMAS probe placed in a Bruker-Biospin Avance III 500 MHz NMR spectrometer. The 13C chemical shifts of Form 1 are shown in FIG. 10.

[0097] Form 1 was analyzed by isothermal vapor sorption analysis, a gravimetric method that measures how fast and how much a sample absorbs a solvent such that the dry powder absorbs water. This analysis is performed by changing the vapor concentration around the sample and measuring the resulting mass change. The isothermal vapor sorption analysis of Form 1 indicates that Form 1 is anhydrous and has a reversible weight gain of less than 0.25% at ambient temperature up to 90% relative humidity.

[0098] Form 4 Form 4 is a crystalline, non-hygroscopic, anhydrous form of the compound of Formula I that can be produced as described in Example 2.

[0099] Form 4 was characterized by the pattern of PXRD shown in FIG. 3, which was measured with a PANalytical X’Pert PRO MPD using an incident beam of Cu radiation generated using an Optix long high-precision focused source. The PXRD pattern of Form 4, represented by degree (2θ) and relative intensity with a relative intensity of ≧2.0%, is shown in FIG. 4. The relative intensity may vary depending on the crystal size and form.

[0100] Form 4 was also characterized by the pattern of the Raman spectrum shown in FIG. 7, which was performed with an NXR FT-Raman module connected to a Nexus 670 FT-IR spectrophotometer (Thermo Nicolet) equipped with an InGaAs detector. The peaks of the Raman spectrum of Form 4 are shown in FIG. 8.

[0101] Also, Form 4 was characterized by the pattern of the solid-state NMR spectrum shown in FIG. 11, which was performed with a Bruker-Biospin 4 mm CPMAS probe placed in a Bruker-Biospin Avance III 500 MHz NMR spectrometer. The 13C chemical shifts of Form 4 are shown in FIG. 12.

[0102] Form 2 Form 2 is a crystalline THF solvate of the compound of formula I that can be produced as described in Example 3.

[0103] The calculated powder pattern of Form 2 shown in FIG. 13 was prepared using Mercury v.3.1 (http: / / www.ccdc.cam.ac.uk / mercury / ).

[0104] Form 6 Form 6 is a crystalline, non-hygroscopic, anhydrous form of the compound of formula I that can be produced as described in Example 4.

[0105] Form 6 was characterized by the pattern of the PXRD shown in FIG. 14, which was measured with a PANalytical X’Pert PRO MPD using an incident beam of Cu radiation generated using an Optix long high-precision focused source. The PXRD pattern of Form 6, represented by degrees (2θ) and relative intensities with a relative intensity of ≧ 2.0%, is shown in FIG. 15. The relative intensity can vary depending on the crystal size and form.

[0106] Form 6 was also characterized by the pattern of the Raman spectrum shown in FIG. 16, which was performed with an NXR FT-Raman module connected to a Nexus 670 FT-IR spectrophotometer (Thermo Nicolet) equipped with an InGaAs detector. The peaks of the Raman spectrum of Form 6 are shown in FIG. 17.

[0107] In addition, Form 6 was characterized by the pattern of the solid-state NMR spectrum shown in Fig. 18, which was performed using a Bruker-Biospin 4mm CPMAS probe placed in a Bruker-Biospin Avance III 500 MHz NMR spectrometer. The 13C chemical shifts of Form 6 are shown in Fig. 19.

[0108] Pharmaceutical composition The active agent of the present invention (i.e., the solid form of the compound of formula I described herein) can be formulated into a pharmaceutical composition suitable for mammalian medical use. Using any suitable route of administration, an effective dose of any of the solid forms of the compound of formula I described herein can be provided to a patient. For example, oral or parenteral formulations can be used. Dosage forms include capsules, tablets, dispersions, suspensions, etc., such as enteric-coated capsules and / or tablets, capsules and / or tablets containing enteric-coated pellets of the solid form of the compound of formula I described herein. In all dosage forms, the solid form of the compound of formula I described herein can be mixed with other suitable constituent substances. The composition is conveniently provided in unit dosage form and can be prepared by any method known in the pharmaceutical art. The pharmaceutical composition of the present invention comprises a therapeutically effective amount of the active agent and one or more inert pharmaceutically acceptable carriers, and optionally any other therapeutic components, stabilizers, etc. The carrier must be pharmaceutically acceptable in the sense that it is compatible with the other components of the formulation and not unduly harmful to its recipient. The composition can further comprise diluents, buffers, binders, disintegrants, thickeners, lubricants, preservatives (including antioxidants), flavoring agents, flavor masking agents, inorganic salts (e.g., sodium chloride), antibacterial agents (e.g., benzalkonium chloride), sweetening agents, antistatic agents, surfactants (e.g., polysorbates such as "TWEEN20 (trademark)" and "TWEEN80 (trademark)", and Pluronic (registered trademark) F68 and F88 available from BASF), sorbitan esters, lipids (e.g., phospholipids such as lecithin and other phosphatidylcholines, phosphatidylethanolamine, fatty acids and fatty acid esters, steroids (e.g., cholesterol)), and chelating agents (e.g., EDTA, zinc, and other such suitable cations).Other pharmaceutical additives and / or adjuncts suitable for use in the compositions according to the invention are listed in Remington: The Science & Practice of Pharmacy, 19th Edition, Williams & Williams, (1995), and "Physician's Desk Reference", 52nd Edition, Medical Economics, Montvale, NJ (1998), and "Handbook of Pharmaceutical Excipients", 3rd Edition, edited by A.H. Kibbe, Pharmaceutical Press, 2000. The active agents of the present invention can be formulated into compositions including those suitable for oral, rectal, topical, nasal, ophthalmic, or parenteral (including intraperitoneal, intravenous, subcutaneous, or intramuscular injection) administration.

[0109] The amount of the active agent in the formulation varies depending on various factors including the dosage form, the condition being treated, the target patient population, and other considerations, and is usually readily determined by those skilled in the art. A therapeutically effective amount is the amount necessary to inhibit transthyretin (TTR) dissociation (i.e., prevent dissociation of the native TTR tetramer into monomers). The composition usually contains an active agent in the range of about 0.001 wt% to about 99 wt%, preferably about 0.01 wt% to about 5 wt%, more preferably about 0.01 wt% to 2 wt%, and also depends on the relative amounts of the additives / adjuncts contained in the composition.

[0110] The pharmaceutical compositions of the present invention are administered in conventional dosage forms prepared by combining a therapeutically effective amount of the active agent as an active ingredient with one or more suitable pharmaceutical carriers according to conventional procedures. These procedures may require mixing, granulating, and compressing, or dissolving the ingredients as appropriate for the desired preparation.

[0111] The pharmaceutical carrier to be used may be solid or liquid. Exemplary solid carriers include lactose, sucrose, talc, gelatin, agar, pectin, gum arabic, magnesium stearate (magnesiun), stearic acid, etc. Exemplary liquid carriers include syrup, peanut oil, olive oil, water, etc. Similarly, the carrier can contain alone or together with wax, ethyl cellulose, hydroxypropyl methyl cellulose, methyl methacrylate, etc. materials known in the art such as glyceryl monostearate or glyceryl distearate with time delay or time release properties.

[0112] Various pharmaceutical forms can be used. Thus, when using a solid carrier, the preparation can be tableted, put into hard gelatin capsules in powder or pellet form, or in the form of troches or lozenges. The amount of the solid carrier may vary, but is usually about 25 mg to about 1 g. When using a liquid carrier, the preparation can be in the form of syrup, emulsion, soft gelatin capsules, sterile injection or suspension in ampoules or vials, or non-aqueous liquid suspensions.

[0113] The actual dosage of the solid form of the compound of formula I described herein for use in the compositions of the present invention will be understood to vary depending on the particular solid form used, the particular composition being formulated, the mode of administration, and the particular site, host, and disease being treated. Taking into account the experimental data regarding the agent and using conventional dosage determination tests, one of ordinary skill in the art can determine the optimal dosage for a given set of circumstances. For oral administration, exemplary daily dosages commonly used are about 0.001 to about 1000 mg / kg body weight, more preferably about 0.001 to about 50 mg / kg body weight, with the treatment course being repeated at appropriate intervals. Administration of prodrugs is generally carried out at weight levels that are chemically equivalent to the weight levels of the fully active form. In the practice of the present invention, the most appropriate route of administration, as well as the scale of the therapeutic dose, will depend on the nature and severity of the disease to be treated. Dosage and frequency of administration may also vary according to the age, weight, and response of the individual patient. Typically, suitable oral dosage forms can cover a dosage range of 0.5 mg to 100 mg of the total daily dose of the active ingredient administered in a single dose or divided into equal portions. The preferred amount of the solid form of the compound of formula I described herein in such a formulation is from about 0.5 mg to about 20 mg, for example, from about 1 mg to about 10 mg or from about 1 mg to about 5 mg, etc.

[0114] The compositions of the present invention can be manufactured in a manner commonly known for preparing pharmaceutical compositions, using conventional techniques such as, for example, mixing, dissolving, granulating, emulsifying, encapsulating, entrapping, or lyophilizing. The pharmaceutical compositions can be formulated in a conventional manner using one or more physiologically acceptable carriers selected from additives and adjuvants that facilitate the processing of the active compound into a pharmaceutically usable preparation.

[0115] For oral administration, the solid forms of the compounds of Formula I described herein can be formulated by combining them with pharmaceutically acceptable carriers known in the art and an active agent. Such carriers can formulate the compounds of the present invention into tablets, pills, capsules, gels, syrups, slurries, suspensions, etc. for oral ingestion by the patient to be treated. Solid additives can be used in combination with the active agent, and optionally, the resulting mixture can be pulverized, and after adding appropriate adjuvants, the mixture of granules can be processed to obtain a pharmaceutical preparation for oral use. Suitable additives include fillers such as sugars including lactose, sucrose, mannitol, or sorbitol; and cellulose preparations such as corn starch, wheat starch, rice starch, potato starch, gelatin, gum, methylcellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, or polyvinylpyrrolidone (PVP). Optionally, disintegrants such as cross-linked polyvinylpyrrolidone, agar, or salts thereof such as alginic acid or sodium alginate can be added.

[0116] Pharmaceutical preparations that can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer such as glycerin or sorbitol. Push-fit capsules can contain an active ingredient mixed with a filler such as lactose, a binder such as starch, and / or a lubricant such as talc or magnesium stearate, and optionally a stabilizer. In soft capsules, the active agent can be dissolved or suspended in a suitable liquid such as fatty oil, liquid paraffin, or liquid polyethylene glycol. Additionally, a stabilizer can be added. All formulations for oral administration should be in dosages suitable for such administration. For buccal administration, the composition can take the form of tablets or lozenges formulated in a conventional manner.

[0117] For ocular administration, the solid form of the compounds of Formula I described herein can be delivered in a pharmaceutically acceptable ocular vehicle, whereby the compound is in contact with the ocular surface for a sufficient period of time to penetrate into the corneal and internal regions of the eye, including, for example, the anterior chamber, posterior chamber, vitreous, aqueous humor, vitreous humor, cornea, iris / ciliary body, lens, choroid / retina, and sclera. The pharmaceutically acceptable ocular vehicle can be, for example, an ointment, vegetable oil, or encapsulating material. The active agent of the present invention can also be injected directly into the vitreous humor, aqueous humor, or subtenon.

[0118] Alternatively, the active ingredient can be in the form of a powder for constitution with a suitable vehicle, for example, sterile pyrogen-free water, before use. The solid forms of the compounds of Formula I described herein can also be formulated into rectal or vaginal compositions such as suppositories or retention enemas containing conventional suppository bases such as cocoa butter or other glycerides.

[0119] In addition to the above-described formulations, the solid forms can also be formulated as depot preparations. Such long-acting formulations can be administered by implantation (e.g., subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the solid forms can be formulated with a suitable polymer or hydrophobic material (e.g., as an emulsion in an acceptable oil) or ion exchange resin or as a slightly insoluble derivative, for example, as a slightly insoluble salt.

[0120] Furthermore, the solid forms of the compounds of Formula I described herein can be delivered using sustained release systems such as semipermeable matrices of solid hydrophobic polymers containing a therapeutic agent. A variety of sustained release materials are established and are known to those of skill in the art.

[0121] The pharmaceutical compositions can also contain suitable solid or gel phase carriers or additives. Examples of such carriers or additives include polymers such as calcium carbonate, calcium phosphate, sugars, starches, cellulose derivatives, gelatin, and polyethylene glycol.

[0122] In certain embodiments, the present invention relates to any of the foregoing pharmaceutical compositions, wherein the solid form is Form 1. In certain embodiments, the present invention relates to any of the foregoing pharmaceutical compositions, wherein the solid form is Form 4.

[0123] Manufactured article The solid forms of the compounds of Formula I described herein can be packaged as a manufactured article containing a packaging material and a solid form of a compound of Formula I provided herein that is effective for modulating TTR folding or for treating, preventing, or ameliorating one or more symptoms of a TTR-mediated disease or disorder, or a disease or disorder in which TTR misfolding is involved, and a label indicating that this solid form is used for modulating TTR folding or for treating, preventing, or ameliorating one or more symptoms of a TTR-mediated disease or disorder, or a disease or disorder in which TTR misfolding is involved.

[0124] The manufactured articles provided in the present invention contain a packaging material. Packaging materials used in the packaging of pharmaceutical products are well known to those skilled in the art. See, for example, U.S. Pat. Nos. 5,323,907, 5,052,558, and 5,033,252. Examples of pharmaceutical packaging materials include, but are not limited to, blister packs, bottles, tubes, inhalers, pumps, bags, vials, containers, syringes, bottles, and any packaging material suitable for the selected formulation and the intended mode of administration and treatment, which treatment includes various treatments for any disease or disorder in which TTR misfolding is involved as an intervening or causative agent in the symptoms or cause.

[0125] In certain embodiments, the present invention relates to any of the aforementioned articles of manufacture, wherein the solid form is Form 1. In certain embodiments, the present invention relates to any of the aforementioned articles of manufacture, wherein the solid form is Form 2. In certain embodiments, the present invention relates to any of the aforementioned articles of manufacture, wherein the solid form is Form 4. In certain embodiments, the present invention relates to any of the aforementioned articles of manufacture, wherein the solid form is Form 6.

[0126] in vitro biological assay Using several in vitro tests, the solid forms can be evaluated for their ability to stabilize transthyretin tetramers or to prevent fibril formation. These tests can include fibril formation assays, plasma selectivity assays, determination of the three-dimensional structure of the transthyretin compound complex (e.g., by X-ray crystallography), kinetics of transthyretin tetramer dissociation or fibril formation, and determination of the stoichiometry and energy of the transthyretin compound interaction, e.g., by centrifugation or calorimetry. Details of exemplary in vitro assays are provided in U.S. Patent No. 7,214,695, which is hereby incorporated by reference in its entirety.

[0127] Methods of using the solid forms of the present invention The compounds of Formula I described herein are useful for the stabilization of the protein transthyretin (TTR), the dissociation of which is involved in TTR amyloidosis (i.e., preventing the dissociation of native TTR tetramers into monomers, thereby inhibiting TTR amyloid fibril formation), and thus provide for the treatment of transthyretin amyloid diseases in mammals, including humans.

[0128] At least some amyloid diseases appear to be caused by the deposition of any one of more than 20 non-homologous proteins or protein fragments, ultimately resulting in a protofibrillar cross-β-sheet quaternary structure. The formation of amyloid fibrils from a normally folded protein such as transthyretin requires protein misfolding that generates assembly-competent intermediates. The process of transthyretin (TTR) amyloid formation appears to cause senile systemic amyloidosis (SSA), familial amyloidotic polyneuropathy (FAP), and familial amyloid cardiomyopathy (FAC). SSA is associated with the deposition of wild-type TTR, while FAP and FAC are caused by amyloid formation of one of more than 80 TTR variants. See, for example, Colon, W.; Kelly, J.W., Biochemistry, 1992, 31, 8654-60; Kelly, J.W., Curr. Opin. Struct. Biol., 1996, 6, 11-7; Liu, K. et al., Nat. Struct. Biol., 2000, 7, 754-7; Westermark, P. et al., Proc. Natl. Acad. Sci. U.S.A., 1990, 87, 2843-5; Saraiva, M.J. et al., J. Clin. Invest., 1985, 76, 2171-7; Jacobson, D.R. et al., N. Engl. J. Med., 1997, 336, 466-73; Buxbaum, J.N., Tagoe, C.E., Ann. Rev. Med., 2000, 51, 543-569; and Saraiva, M.J., Hum. Mutat., 1995, 5, 191-6, each of which is incorporated by reference in its entirety. Another TTR amyloid disease includes cardiac amyloidosis after liver transplantation, peripheral nerve amyloidosis after liver transplantation, leptomeningeal amyloidosis, carpal tunnel syndrome associated with a transthyretin mutant, vitreous deposition, and skin amyloidosis associated with a transthyretin mutant.

[0129] A therapeutically effective amount of the compound of formula 1 is generally administered in the form of a pharmaceutical composition to treat diseases mediated by modulation or regulation of TTR dissociation. "Effective amount" is intended to mean an amount of the agent sufficient to effect treatment of a disease mediated by TTR dissociation when administered to a mammal in need of such treatment. Thus, a therapeutically effective amount of Compound 1 is an amount sufficient to modulate, regulate, or inhibit the dissociation of TTR and reduce or alleviate the disease state mediated by its activity. "Treat" is intended to mean at least alleviation of a disease state in a mammal such as a human, which includes preventing the occurrence of a disease state in the mammal, particularly when it has been found that the mammal is prone to having the disease state but has not yet been diagnosed as having the disease state; modulating and / or inhibiting the disease state; and / or reducing the disease state. Exemplary disease states include senile systemic amyloidosis (SSA), familial amyloid polyneuropathy (FAP), familial amyloid cardiomyopathy (FAC), cardiac amyloidosis after liver transplantation, peripheral nerve amyloidosis after liver transplantation, leptomeningeal amyloidosis, carpal tunnel syndrome associated with a transthyretin mutant, vitreous deposition, and skin amyloidosis associated with a transthyretin mutant.

Example

[0130] The following examples illustrate separate forms of the invention, namely Forms 1 and 4, which do not limit the scope of the invention as defined herein or claimed below.

[0131] (Example 1) Preparation of Form 1 4-Amino-3-hydroxybenzoic acid (1.0 equivalent, LR) was dissolved at 20 °C in a mixture of tetrahydrofuran (19 L / kg) and water (1.9 L / Kg). 3,5-Dichlorobenzoyl chloride (1.3 equivalents) was added as a tetrahydrofuran solution (1.9 L / kg), and the mixture was stirred at 20 °C for at least 30 minutes. When the completion of the reaction was judged by HPLC (residual 4-amino-3-hydroxybenzoic acid < 5%), triethylamine (1.2 equivalents) was added, the mixture was heated to 35 °C, and stirred for at least 90 minutes. The solvent was partially replaced with ethanol by a certain level of distillation until the remaining THF was 5 - 15%. The slurry was cooled to 20 °C, stirred for at least 60 minutes, and then the slurry was filtered. The solid was washed with ethanol (3 × 4 L / kg) and then vacuum dried at 65 °C for at least 16 hours to obtain pure 4-[(3,5-dichlorobenzoyl)amino]-3-hydroxybenzoic acid in a yield of 88 - 92%.

[0132] To a slurry of tetrahydrofuran (10 L / kg) containing 4-[(3,5-dichlorobenzoyl)amino]-3-hydroxybenzoic acid (1.0 equivalent), water (4 equivalents) was added following triethylamine (1.1 equivalents). The mixture was held at 20 - 25 °C for 1 hour, and then the mixture was filtered to remove all remaining insoluble materials. Methanesulfonic acid (1.6 equivalents) was added to form a slurry. The THF / water was replaced with toluene at a certain level until the reaction temperature reached at least 107 °C, the replacement was stopped at 107 °C, and then the reaction was refluxed for at least 15 hours. When the completion of the reaction was judged by UPLC, i.e., when the purity reached > 95%, it was cooled to 20 °C and 2-propanol (5 L / kg) was added. The slurry was granulated for at least 60 minutes, then filtered, washed twice with 2-propanol (4 L / kg for each wash), and vacuum dried at 60 - 70 °C for at least 18 hours to obtain Form 1 in a yield of 82 - 89%.

[0133] (Example 2) Preparation of Form 4 Form 1 (187 mg) was suspended in tetrahydrofuran (7.5 mL), and the suspension was heated at 75 °C. The clear solution was filtered while warm through a pre-warmed 0.2-μm nylon filter and placed into a container containing toluene (25 mL) cooled in an ice / water bath. The sample was stored overnight in a freezer (-10 to -25 °C). Form 4 was collected by vacuum filtration while cooling.

[0134] (Example 3) Preparation of Form 2 A 3 mg / mL THF solution of Form 1 was evaporated under ambient conditions in a hood to obtain crystals. The following results were shown from single crystal analysis: Empirical formula C 14 H7NO3Cl2 Formula weight 308.12 Temperature Ambient Wavelength 1.54178 Å Crystal system Triclinic Space group P-1 Unit cell dimensions a = 3.7740(2) Å α = 80.668(3)° b = 13.6536(8) Å β = 89.381(4)° c = 15.5098(9) Å γ = 89.520(3)° Volume 788.56(8) Å 3 Z 4 Density (calculated) 1.365 Mg / m 3 F 2 goodness-of-fit 1.112 Final R indices [I>2sigma(I)] R1 = 0.0776, wR2 = 0.2360 R indices (all data) R1 = 0.1026, wR2 = 0.2561

[0135] (Example 4) Preparation of Form 6 Form 1 (4168 mg) was suspended in tetrahydrofuran (100 mL) and heated with stirring at 60 °C. Dimethylacetamide (5 mL) was added. The resulting solution was filtered through a pre-warmed 0.2 μm nylon filter while keeping warm and placed into a container containing dichloromethane cooled in an ice / water bath. The observed solid was isolated by vacuum filtration and air-dried at ambient temperature.

[0136] (Example 5) Preparation of amorphous 6-carboxy-2-(3,5-dichlorophenyl)-benzoxazole Form 1 (79.7 mg) was suspended in 5 mL of dioxane / water (80 / 20) and heated at about 80 °C. The resulting clear solution was filtered through a pre-warmed 0.2 μm nylon filter while keeping warm and placed into a pre-warmed receiving vial. The sample was then frozen in a dry ice / IPA bath and transferred to a lyophilizer and left for 2 days. The solid was collected.

Claims

1. A crystalline form of 6-carboxy-2-(3,5-dichlorophenyl)-benzoxazole, having a solid NMR spectrum containing 13C chemical shifts (ppm) at 120.8 ± 0.2 and 127.7 ± 0.

2.

2. The crystalline form of 6-carboxy-2-(3,5-dichlorophenyl)-benzoxazole according to Claim 1, wherein the solid NMR spectrum further contains a 13C chemical shift (ppm) at 139.6 ± 0.

2.

3. The crystalline form of 6-carboxy-2-(3,5-dichlorophenyl)-benzoxazole according to Claim 1, wherein the solid NMR spectrum further contains a 13C chemical shift (ppm) at 144.7 ± 0.

2.

4. A crystalline form of 6-carboxy-2-(3,5-dichlorophenyl)-benzoxazole, having a powder X-ray diffraction pattern containing a peak at a diffraction angle (2θ) of 28.6 ± 0.

2.

5. The crystalline form of 6-carboxy-2-(3,5-dichlorophenyl)-benzoxazole according to Claim 4, wherein the powder X-ray diffraction pattern further contains peaks at diffraction angles (2θ) of 16.5 ± 0.2 and 26.7 ± 0.

2.

6. The crystalline form of 6-carboxy-2-(3,5-dichlorophenyl)-benzoxazole according to Claim 4, wherein the powder X-ray diffraction pattern further contains peaks at diffraction angles (2θ) of 15.4 ± 0.2 and 20.2 ± 0.

2.

7. The crystalline form of 6-carboxy-2-(3,5-dichlorophenyl)-benzoxazole according to Claim 6, wherein the powder X-ray diffraction pattern further contains a peak at a diffraction angle (2θ) of 29.0 ± 0.

2.

8. The crystalline form of 6-carboxy-2-(3,5-dichlorophenyl)-benzoxazole according to Claim 7, wherein the powder X-ray diffraction pattern further contains a peak at a diffraction angle (2θ) of 23.5 ± 0.

2.

9. A crystalline form of 6-carboxy-2-(3,5-dichlorophenyl)-benzoxazole, having a Raman spectrum containing a Raman shift peak (cm-1) at 1292 ± 2.

10. The crystalline form of 6-carboxy-2-(3,5-dichlorophenyl)-benzoxazole according to claim 9, having a Raman spectrum further comprising Raman shift peaks (cm-1) at 994±2, 1273±2, and 1615±2.

11. The crystalline form of 6-carboxy-2-(3,5-dichlorophenyl)-benzoxazole according to claim 6, wherein the Raman spectrum further comprises Raman shift peaks (cm-1) at 287±2 and 869±2.

12. The crystalline form of 6-carboxy-2-(3,5-dichlorophenyl)-benzoxazole according to claim 11, wherein the Raman spectrum further comprises a Raman shift peak (cm-1) at 213±2.

13. A crystalline form of 6-carboxy-2-(3,5-dichlorophenyl)-benzoxazole, having a powder X-ray diffraction pattern comprising peaks at diffraction angles (2θ) of 26.7±0.2 and 28.6±0.2, and a solid NMR spectrum comprising a 13C chemical shift (ppm) at 127.7±0.

2.

14. A crystalline form of 6-carboxy-2-(3,5-dichlorophenyl)-benzoxazole, having a Raman spectrum comprising Raman shift peaks (cm-1) at 1292±2 and 1615±2, and a solid NMR spectrum comprising a 13C chemical shift (ppm) at 127.7±0.

2.

15. The crystalline form according to any one of claims 1 to 14, which is non-hygroscopic and anhydrous.

16. The crystalline form according to any one of claims 1 to 15, which is substantially pure.

17. A pharmaceutical composition comprising a therapeutically effective amount of the crystalline form according to any one of claims 1 to 16, mixed with at least one pharmaceutically acceptable additive.

18. A method for treating transthyretin amyloidosis in a mammal, comprising administering to the mammal a therapeutically effective amount of the crystalline form according to any one of claims 1 to 16 or the pharmaceutical composition according to claim 17.

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