Solid state forms of n-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide

Abstract

The invention discloses a solid state form of N-(2-(4-cyano thiazolidine-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide, and a preparation method of the solid state form. A corresponding pharmaceutical composition; their use in the treatment or prevention of prolyl endopeptidase fibroblast activation protein (FAP) mediated conditions; a kit; and a preparation method thereof.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit under 35 U.S.C. §119(e) of U.S. Provisional Application No. 63 / 366,696, filed on June 21, 2022. The above-listed applications are incorporated by reference in their entirety for all purposes. Technical Field

[0003] The present disclosure generally relates to solid state forms of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide, including crystalline forms of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide. The present disclosure also relates to pharmaceutical compositions comprising a crystalline form of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide; uses of pharmaceutical compositions comprising a crystalline form of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide for treating or preventing disorders mediated by prolyl endopeptidase fibroblast activation protein (FAP); kits comprising a pharmaceutical composition comprising a crystalline form of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide; and methods for preparing a crystalline form of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide. Background Art

[0004] FAP (type II transmembrane serine protease) is expressed by fibroblast-like cells involved in tissue remodeling and healing. In the context of non-alcoholic steatohepatitis (NASH), FAP is upregulated on the cell surface of activated hepatic stellate cells involved in fibrosis formation (Hepatology 1999, 29, 1768), which is a major aspect of predicting disease outcomes in NASH (Gastroenterology 2020, 158, 1611). FAP can also exist as a shed plasma protease. Increased circulating FAP levels are associated with NASH disease severity (Diabetes Res Clin Pract 2015, 108, 466).

[0005] FAP has a common cleavage motif after Gly-Pro and exhibits both endopeptidase activity and exopeptidase activity. Known enzyme activities include collagen cleavage (Hepatology 1999, 29, 1768), α2-antiplasmin (α2AP) (Blood 2004 103, 3783) and fibroblast growth factor 21 (FGF21) (Biochem J 2016, 473, 605). FAP activity (including collagen cleavage) at the cell surface of activated fibroblasts generates a pro-fibrotic environment. FAP cleavage of α2AP cross-links α2AP with fibrin more effectively and results in reduced fibrin clearance. FAP cleavage of FGF21 inactivates FGF21 metabolism (Biochem J 2016, 473, 605). All of these activities are associated with the deterioration of NASH disease, and inhibition of FAP has the potential to treat NASH and other conditions by affecting multiple mechanisms.

[0006] Inhibition of FAP activity is a currently undeveloped therapeutic approach for the treatment of NASH and other diseases associated with this activity. There are currently no approved pharmacological agents that inhibit FAP activity generally or specifically. Therefore, there is a need for FAP inhibitors, particularly FAP inhibitors that have pharmacologically suitable selectivity and bioavailability and have physical properties suitable for the manufacture of drug substances and formulation of corresponding drug products.

[0007] The present disclosure addresses this great unmet need by providing solid-state forms of a FAP inhibitor (N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide) that are useful in pharmaceutical compositions and methods for treating FAP-mediated disorders such as NASH. Summary of the invention

[0008] In one aspect, the present disclosure provides a solid state form of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0009] In another aspect, the present disclosure provides a crystalline form of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0010] In another aspect, the present disclosure provides crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Form A.

[0011] In another aspect, the present disclosure provides crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Form B.

[0012] In another aspect, the present disclosure provides a crystalline form of (R,S)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0013] In another aspect, the present disclosure provides crystalline (R,S)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Type 2.

[0014] In another aspect, the present disclosure provides a pharmaceutical composition comprising a crystalline form of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide and one or more pharmaceutically acceptable excipients.

[0015] In another aspect, the present disclosure provides a method of treating or preventing a disorder mediated by the prolyl endopeptidase fibroblast activation protein (FAP) by administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0016] In another aspect, the present disclosure provides use of a pharmaceutical composition comprising a crystalline form of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide for treating or preventing a disorder mediated by prolyl endopeptidase fibroblast activation protein (FAP).

[0017] In another aspect, the present disclosure provides the use of a crystalline form of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide for the manufacture of a medicament for treating or preventing a disorder mediated by prolyl endopeptidase fibroblast activation protein (FAP).

[0018] In another aspect, the present disclosure provides a kit comprising a pharmaceutical composition comprising a crystalline form of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0019] In another aspect, the present disclosure provides a process for preparing a crystalline form of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 : is a representative transmission powder X-ray diffraction pattern of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Form A.

[0021] Figure 2 A representative solid sample of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide is shown. 13 C NMR spectrum, this sample was approximately 95 wt % Form A and 5 wt % Form B.

[0022] Figure 3 A representative solid sample of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide is shown. 13 C NMR spectrum, this sample is approximately 30 wt % Form A and 70 wt % Form B.

[0023] Figure 4 A representative solid sample of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide is shown. 13 C NMR spectrum showed that the sample was Form B with a small amount of Form A.

[0024] Figure 5 is based on Figure 2 , Figure 3 and Figure 4 The solid state spectra of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Form A and Form B are shown. 13 Comparison of C NMR spectra.

[0025] Figure 6 is a representative differential scanning calorimetry curve of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Form A.

[0026] Figure 7 is a representative thermogravimetric analysis thermogram of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Form A.

[0027] Figure 8 is a representative gravimetric vapor sorption curve of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Form A.

[0028] Fig. 9: is a representative transmission powder X-ray diffraction pattern of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Form B.

[0029] Fig.10 is based on Figure 1 and Fig. 9 Comparison of transmission powder X-ray diffraction patterns of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Form A and Form B. The diffraction pattern of Form A is shown in Fig.10 The top of the Form B diffraction pattern is Fig.10 bottom.

[0030] Fig.11 is based on Figure 1 and Fig. 9 Comparison of the transmission powder X-ray diffraction patterns of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Form A and Form B, shown in the diffraction pattern, which is used to cover the region of the unique Form B peak. The Form A diffraction pattern is Fig.11 The top of the Form B diffraction pattern is Fig.11 bottom.

[0031] Fig.12 is a representative differential scanning calorimetry curve of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Form B.

[0032] Fig.13 is a representative thermogravimetric analysis thermogram of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Form B.

[0033] Fig.14 is a representative gravimetric vapor sorption plot of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Form B.

[0034] Fig.15 is a representative reflection powder X-ray diffraction pattern of crystalline (R,S)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Type 2.

[0035] Fig.16 is a representative differential scanning calorimetry curve of crystalline (R,S)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Type 2.

[0036] Fig.17 is a representative gravimetric vapor sorption curve of crystalline (R,S)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Type 2.

[0037] Fig.18 is a representative reflection powder X-ray diffraction pattern of amorphous (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0038] Fig.19 is a representative differential scanning calorimetry curve of amorphous (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0039] Fig. 20 is a representative gravimetric vapor sorption curve for amorphous (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0040] Fig.21 Representative solubility curves of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Form B in acetonitrile and methanol are shown. DETAILED DESCRIPTION

[0041] Many embodiments are described in detail throughout the specification, and these embodiments are obvious to those skilled in the art. Such embodiments are provided by way of example only, and are not intended to limit the scope of the invention in other ways. Various alternatives to the embodiments may be adopted when practicing the present invention.

[0042] I. Definitions

[0043] With respect to the embodiments disclosed in this specification, the following terms have the meanings set forth below:

[0044] Reference to "a" or "an" means "one or more." Throughout, except for numerical indications, plural and singular should be regarded as interchangeable.

[0045] When ranges are used herein to describe, for example, physical or chemical properties, all combinations and subcombinations of ranges and specific embodiments therein are intended to be included. When referring to a number or a numerical range, the use of the term "about" or "approximately" means that the number or numerical range referred to is an approximation within experimental variability (or within statistical experimental error), and therefore the number or numerical range will vary between 1% and 15% of the number or numerical range.

[0046] Unless the context requires otherwise, the words "comprises" or "includes" are used with the clear understanding that they are to be interpreted as inclusive rather than exclusive, and applicant intends that each of these words be so interpreted when interpreting this patent, including the claims below.

[0047] The term "amorphous form" refers to a form of a compound that lacks long range crystalline order.

[0048] As used herein, the terms "co-administered," "administered in combination with," and "administered in conjunction with" encompass administering two or more agents to a subject such that both agents and / or their metabolites are present in the subject at the same time. Co-administration includes simultaneous administration in separate compositions, administration at different times in separate compositions, or administration in a composition in which two or more agents are present.

[0049] The term "crystalline form" is intended to include all crystalline forms of a compound, including, for example, polymorphs, pseudopolymorphs, solvates, hydrates, unsolvated polymorphs (including anhydrates), and conformational polymorphs, and mixtures thereof, unless a specific crystalline form is mentioned.

[0050] The term "therapeutically effective amount" of a pharmacological agent is an amount sufficient to achieve a beneficial or desired result (including a clinical result), and will therefore depend on the context in which it is administered. For example, when a pharmacological agent is administered to treat liver disease, a therapeutically effective amount of the agent is an amount of the agent sufficient, alone or in combination with an additional therapy, to provide an anti-liver disease effect in a subject as compared to the response obtained without administration of the agent.

[0051] The term "pharmaceutically acceptable" is used as an adjective in this specification to mean that the modified noun is suitable for use as a pharmaceutical product or as a part of a pharmaceutical product. For example, the term "pharmaceutical carrier" or "pharmaceutically acceptable excipient" is intended to include any and all carriers or excipients suitable for use in mammals, particularly humans.

[0052] When used in connection with powder X-ray diffraction, the terms "reflection" or "reflection mode" refer to the reflection (also known as Bragg-Brentano) sampling mode.

[0053] The term "prevent" is readily understood by the ordinarily skilled physician and, with respect to treatment of a particular condition, may include primary prevention, which is intended to have its normal meaning, and includes preventing the development of the condition, as well as secondary prevention, whereby the condition has already developed and the patient is temporarily or permanently protected from an exacerbation or worsening of the disease or the development of new symptoms associated with the condition.

[0054] The term "solvate" refers to a crystalline phase of a compound in physical association with one or more molecules of a solvent. A crystalline phase of a compound in physical association with one or more molecules of water is called a "hydrate."

[0055] When used in conjunction with powder X-ray diffraction, the terms "transmission" or "transmission mode" refer to the transmission (also known as Debye-Scherrer) sampling mode.

[0056] The term "treat" is readily understood by an ordinarily skilled physician and, with respect to treatment of a particular condition, may include (1) reducing the extent or cause of the condition being treated, and / or (2) alleviating or ameliorating one or more symptoms associated with the condition. Treatment of liver disease may include, for example, stabilizing (i.e., not worsening), delaying or slowing the spread or progression of liver disease; prolonging survival compared to expected survival without treatment; and / or otherwise ameliorating or alleviating the severity of liver disease in whole or in part.

[0057] As used herein, "enantiomeric purity" refers to the relative amount (expressed as a percentage) of a particular enantiomer present relative to the other enantiomer. For example, if a compound that can potentially have either (R)- or (S)-isomer configurations exists as a racemic mixture, the enantiomeric purity relative to the (R)- or (S)-isomer is about 50%. If the compound has one isomeric form that is superior to the other, e.g., 80% (S)-isomer and 20% (R)-isomer, the enantiomeric purity of the compound relative to the (S)-isomer form is 80%. The enantiomeric purity of a compound can be determined in a variety of ways, including but not limited to chromatography using a chiral support, polarization measurements of polarized light rotation, nuclear magnetic resonance spectroscopy using chiral shift reagents (including but not limited to chiral complexes containing lanthanides or Pirkle reagents), or derivatization of the compound using a chiral compound (such as Mosher's acid) followed by chromatography or nuclear magnetic resonance spectroscopy.

[0058] In some embodiments, an enantiomerically enriched composition has a higher potency per unit mass of therapeutic utility than a racemic mixture of the composition. Enantiomers can be separated from the mixture by methods known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred enantiomers can be prepared by asymmetric synthesis. See, for example, Jacques et al., Enantiomers, Racemates and Resolutions, Wiley Interscience, New York, 1981; Eliel, Stereochemistry of Carbon Compounds, McGraw-Hill, NY, 1962; and Eliel and Wilen, Stereochemistry of Organic Compounds, Wiley-Interscience, New York, 1994.

[0059] As used herein, the terms "enantiomerically enriched" and "non-racemic" refer to compositions in which the weight percentage of one enantiomer is greater than the amount of that one enantiomer in a control mixture of a racemic composition (e.g., greater than 1:1 by weight). For example, an enantiomerically enriched preparation of the (R)-enantiomer means a preparation of a compound having greater than 50% by weight (such as at least 75% by weight, or such as at least 80% by weight) of the (R)-enantiomer relative to the (S)-enantiomer. In some embodiments, the enrichment may be significantly greater than 80% by weight, thereby providing a "substantially enantiomerically enriched" or "substantially non-racemic" preparation, which refers to a preparation of a composition having at least 85% by weight (such as at least 90% by weight, or such as at least 95% by weight) of one enantiomer relative to the other enantiomer. The term "enantiomerically pure" or "substantially enantiomerically pure" refers to a composition comprising at least 98% of a single enantiomer and less than 2% of the opposite enantiomer.

[0060] II. Crystalline Form of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Mode

[0061] Compounds that are active pharmaceutical ingredients in drug products may exist in different solid forms that exhibit different physical properties. These differences in physical properties can affect the manufacture and formulation of drug products. These physical properties may include, but are not limited to: (1) bulk properties, such as molar volume, density, and hygroscopicity, (2) thermodynamic properties, such as melting temperature, vapor pressure, and solubility, (3) kinetic properties, such as dissolution rate and stability (including stability under ambient conditions, particularly to moisture and under storage conditions), (4) surface properties, such as surface area, wettability, interfacial tension, and shape, (5) mechanical properties, such as hardness, tensile strength, compressibility, compactability, handling, flow, and mixing; and (6) filtration properties. Therefore, solid forms of compounds (particularly crystalline forms of compounds) that provide improvements in one or more of these physical properties relative to other solid forms of the compound are desirable. Therefore, the discovery of new solid forms of pharmaceutically useful compounds provides potential opportunities to improve the performance characteristics of corresponding drug products and related manufacturing processes.

[0062] The present disclosure provides a solid form of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide. In one aspect, the solid form is a crystalline form. Each crystalline form described in this specification has one or more of the above-mentioned favorable properties relative to one or more of the other solid forms of the compound. In another aspect, the solid form is a crystalline anhydrous. In another aspect, the crystalline form is substantially pure. As used in this specification, the term "substantially pure" means that the crystalline form of the compound comprises at least about 90% by weight of the desired crystalline form relative to any other solid form of the compound. In one aspect, the crystalline form of the compound comprises at least about 95% by weight of the desired crystalline form. In another aspect, the crystalline form of the compound comprises at least about 96% by weight of the desired crystalline form. In another aspect, the crystalline form of the compound comprises at least about 97% by weight of the desired crystalline form. In another aspect, the crystalline form of the compound comprises at least about 98% by weight of the desired crystalline form. In another aspect, the crystalline form of the compound comprises at least about 99% by weight of the desired crystalline form.

[0063] In some embodiments, the present disclosure provides a crystalline form of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide having the following chemical structure:

[0064]

[0065] In some embodiments, the present disclosure provides crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Form A.

[0066] In some embodiments, Form A is characterized by a transmission X-ray powder diffraction pattern: (i) it comprises at least one peak selected from the group consisting of 12.0 ± 0.2 ° 2θ, 18.4 ± 0.2 ° 2θ, 19.8 ± 0.2 ° 2θ, and 21.7 ° 2θ ± 0.2 ° 2θ, and (ii) it does not comprise peaks with intermediate or stronger relative intensities at 18.7 ± 0.2 ° 2θ and 22.4 ° 2θ ± 0.2 ° 2θ. In one aspect, the transmission X-ray powder diffraction pattern does not comprise peaks at 18.7 ± 0.2 ° 2θ and 22.4 ° 2θ ± 0.2 ° 2θ. In another aspect, the transmission X-ray powder diffraction pattern comprises at least two, three, or four peaks selected from the group consisting of 12.0 ± 0.2 ° 2θ, 18.4 ± 0.2 ° 2θ, 19.8 ± 0.2 ° 2θ, and 21.7 ° 2θ ± 0.2 ° 2θ. In another aspect, the transmission X-ray powder diffraction pattern comprises peaks at 12.0 ± 0.2 ° 2θ, 18.4 ± 0.2 ° 2θ, 19.8 ± 0.2 ° 2θ, and 21.7 ° 2θ ± 0.2 ° 2θ. In another aspect, the transmission X-ray powder diffraction pattern further comprises at least one, two, three or four peaks selected from the group consisting of 13.1 ± 0.2 ° 2θ, 14.4 ± 0.2 ° 2θ, 17.5 ± 0.2 ° 2θ, and 21.1 ° 2θ ± 0.2 ° 2θ. In another aspect, the transmission X-ray powder diffraction pattern further comprises peaks at 13.1 ± 0.2 ° 2θ, 14.4 ± 0.2 ° 2θ, 17.5 ± 0.2 ° 2θ, and 21.1 ° 2θ ± 0.2 ° 2θ. In another aspect, the transmission X-ray powder diffraction pattern comprises peaks at 8.9±0.2°2θ, 12.0±0.2°2θ, 13.1±0.2°2θ, 14.4±0.2°2θ, 17.5±0.2°2θ, 17.9±0.2°2θ, 18.4±0.2°2θ, 19.8±0.2°2θ, 20.6±0.2°2θ, 21.1±0.2°2θ, 21.7±0.2°2θ, 23.6±0.2°2θ, 25.6±0.2°2θ, 27.3±0.2°2θ, 31.2±0.2°2θ, 39.9±0.2°2θ, and 42.0±0.2°2θ. In another aspect, the transmission X-ray powder diffraction pattern comprises peaks at 8.9±0.2°2θ, 12.0±0.2°2θ, 13.1±0.2°2θ, 14.4±0.2°2θ, 17.5±0.2°2θ, 17.9±0.2°2θ, 18.4±0.2°2θ, 19.8±0.2°2θ, Figure 1 The transmission X-ray powder diffraction patterns are essentially the same.

[0067] In some embodiments, Form A is characterized by: solid state 13 C NMR spectrum, which comprises at least one peak selected from the group consisting of 166.1±0.2 ppm, 130.7±0.2 ppm, 117.9±0.2 ppm, 108.6±0.2 ppm and 35.2±0.2 ppm. In one aspect, the solid 13The C NMR spectrum contains peaks at 166.1 ± 0.2 ppm, 117.9 ± 0.2 ppm, and 108.6 ± 0.2 ppm. In another aspect, the solid 13 The C NMR spectrum comprises peaks at 166.1 ± 0.2 ppm, 130.7 ± 0.2 ppm, 117.9 ± 0.2 ppm, 108.6 ± 0.2 ppm, and 35.2 ± 0.2 ppm. In another aspect, the solid 13 The C NMR spectrum comprises peaks at 168.0±0.2 ppm, 166.1±0.2 ppm, 147.5±0.2 ppm, 146.4±0.2 ppm, 143.1±0.2 ppm, 139.5±0.2 ppm, 130.7±0.2 ppm, 125±0.2 ppm, 117.9±0.2 ppm, 108.6±0.2 ppm, 67.1±0.2 ppm, 48.0±0.2 ppm, 42.9±0.2 ppm, and 35.2±0.2 ppm. In another aspect, the solid 13 The C NMR spectrum does not comprise at least one peak selected from the group consisting of 166.9 ± 0.2 ppm, 130.2 ± 0.2 ppm, 118.6 ± 0.2 ppm, 117.4 ± 0.2 ppm, 110.0 ± 0.2 ppm, 49.9 ± 0.2 ppm, 46.6 ± 0.2 ppm, and 34.5 ± 0.2 ppm. 13 The C NMR spectrum does not comprise at least five peaks selected from the group consisting of 166.9 ± 0.2 ppm, 130.2 ± 0.2 ppm, 118.6 ± 0.2 ppm, 117.4 ± 0.2 ppm, 110.0 ± 0.2 ppm, 49.9 ± 0.2 ppm, 46.6 ± 0.2 ppm, and 34.5 ± 0.2 ppm. 13 The C NMR spectrum does not contain peaks at 49.9 ± 0.2 ppm and 46.6 ± 0.2 ppm. In another aspect, the solid 13 The C NMR spectrum does not include peaks at 166.9 ± 0.2 ppm, 118.6 ± 0.2 ppm, 110.0 ± 0.2 ppm, 49.9 ± 0.2 ppm, and 46.6 ± 0.2 ppm. In another aspect, the solid 13 The C NMR spectrum does not include peaks at 166.9±0.2 ppm, 130.2±0.2 ppm, 118.6±0.2 ppm, 117.4±0.2 ppm, 110.0±0.2 ppm, 49.9±0.2 ppm, 46.6±0.2 ppm, and 34.5±0.2 ppm.

[0068] In some embodiments, Form A is characterized by a differential scanning calorimetry curve comprising a melting endotherm with an onset temperature between about 165°C and about 180°C. In one aspect, the endotherm comprises a melting endotherm with an onset temperature between about 165°C and about 177°C. In another aspect, the endotherm starts at 171°C ± 5°C. In another aspect, the differential scanning calorimetry curve is Figure 6 The differential scanning calorimetry curves of

[0069] In some embodiments, Form A is characterized by a thermogravimetric analysis thermogram, wherein the crystalline form exhibits a weight loss of less than about 0.5 wt % from about 25°C to about 110°C. In one aspect, the weight loss is less than about 0.2 wt %. In another aspect, the weight loss is less than about 0.1 wt %. In another aspect, the thermogravimetric analysis thermogram is characterized by a weight loss of less than about 0.5 wt % from about 25°C to about 110°C. Figure 7 The thermogravimetric analysis thermograms of the samples were essentially the same.

[0070] In some embodiments, Form A is characterized by a gravimetric vapor sorption curve, wherein the crystalline form exhibits a reversible moisture uptake of less than about 0.5 wt % from about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C. In one aspect, the reversible moisture uptake is less than about 0.2 wt %. In another aspect, the reversible moisture uptake is less than about 0.1 wt %. In another aspect, the gravimetric vapor sorption curve is less than about 0.5 wt %. Figure 8 The weight vapor adsorption curves are essentially the same.

[0071] In some embodiments, Form A is characterized by the physical characterization embodiments described above (transmission X-ray powder diffraction, solid state 13 The method can be characterized by at least two of C NMR, differential scanning calorimetry, thermogravimetric analysis, and / or gravimetric vapor sorption).

[0072] In some embodiments, Form A is characterized by the physical characterization embodiments described above (transmission X-ray powder diffraction, solid state 13 The method can be characterized by at least three of C NMR, differential scanning calorimetry, thermogravimetric analysis, and / or gravimetric vapor sorption).

[0073] In some embodiments, Form A is characterized by the physical characterization embodiments described above (transmission X-ray powder diffraction, solid state 13 The method can be characterized by at least four of C NMR, differential scanning calorimetry, thermogravimetric analysis, and / or gravimetric vapor sorption).

[0074] In some embodiments, Form A is characterized by:

[0075] a transmission X-ray powder diffraction pattern comprising at least one peak selected from the group consisting of 12.0±0.2°2θ, 18.4±0.2°2θ, 19.8±0.2°2θ, and 21.7°2θ±0.2°2θ, and which does not comprise peaks having medium or stronger relative intensities at 18.7±0.2°2θ and 22.4°2θ±0.2°2θ;

[0076] Solid State 13 C NMR spectrum comprising peaks at 166.1±0.2 ppm, 130.7±0.2 ppm, 117.9±0.2 ppm, 108.6±0.2 ppm, and 35.2±0.2 ppm; and

[0077] A differential scanning calorimetry curve comprising a melting endotherm with an onset temperature between about 165°C and about 180°C.

[0078] In some embodiments, Form A is characterized by:

[0079] a transmission X-ray powder diffraction pattern comprising at least one peak selected from the group consisting of 12.0±0.2°2θ, 18.4±0.2°2θ, 19.8±0.2°2θ, and 21.7°2θ±0.2°2θ, and which does not comprise peaks having medium or stronger relative intensities at 18.7±0.2°2θ and 22.4°2θ±0.2°2θ;

[0080] Solid State 13 C NMR spectrum comprising peaks at 166.1±0.2 ppm, 130.7±0.2 ppm, 117.9±0.2 ppm, 108.6±0.2 ppm, and 35.2±0.2 ppm;

[0081] a differential scanning calorimetry curve comprising a melting endotherm with an onset temperature between about 165°C and about 177°C; and

[0082] A thermogravimetric analysis thermogram wherein the crystalline form exhibits a weight loss of less than about 0.2 weight percent from about 25°C to about 110°C.

[0083] In some embodiments, Form A is characterized by:

[0084] a transmission X-ray powder diffraction pattern comprising at least one peak selected from the group consisting of 12.0±0.2°2θ, 18.4±0.2°2θ, 19.8±0.2°2θ, and 21.7°2θ±0.2°2θ, and which does not comprise peaks having medium or stronger relative intensities at 18.7±0.2°2θ and 22.4°2θ±0.2°2θ;

[0085] Solid State 13C NMR spectrum comprising peaks at 166.1±0.2 ppm, 130.7±0.2 ppm, 117.9±0.2 ppm, 108.6±0.2 ppm, and 35.2±0.2 ppm;

[0086] a differential scanning calorimetry curve comprising a melting endotherm with an onset temperature between about 165°C and about 177°C; and

[0087] A gravimetric vapor sorption plot wherein the crystalline form exhibits a reversible moisture uptake of less than about 0.2 wt % from about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C.

[0088] In some embodiments, Form A is characterized by:

[0089] a transmission X-ray powder diffraction pattern comprising at least one peak selected from the group consisting of 12.0±0.2°2θ, 18.4±0.2°2θ, 19.8±0.2°2θ, and 21.7°2θ±0.2°2θ, and which does not comprise peaks having medium or stronger relative intensities at 18.7±0.2°2θ and 22.4°2θ±0.2°2θ;

[0090] Solid State 13 C NMR spectrum comprising peaks at 166.1±0.2 ppm, 130.7±0.2 ppm, 117.9±0.2 ppm, 108.6±0.2 ppm, and 35.2±0.2 ppm;

[0091] a differential scanning calorimetry curve comprising a melting endotherm with an onset at 171°C ± 5°C;

[0092] a thermogravimetric analysis thermogram, wherein the crystalline form exhibits a weight loss of less than about 0.2 weight percent from about 25°C to about 110°C; and

[0093] A gravimetric vapor sorption plot wherein the crystalline form exhibits a reversible moisture uptake of less than about 0.2 wt % from about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C.

[0094] In some embodiments, Form A is a crystalline anhydrate.

[0095] In some embodiments, Form A has an elongated needle-like morphology.

[0096] In some embodiments, Form A is substantially free of any other crystalline forms of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0097] In some embodiments, Form A contains less than 5% by weight of any other crystalline forms of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0098] In some embodiments, Form A contains less than 10% by weight of any other crystalline forms of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0099] In some embodiments, the present disclosure provides crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Form B.

[0100] In some embodiments, Form B is characterized by a transmission X-ray powder diffraction pattern comprising peaks at 18.7 ± 0.2 ° 2θ and 22.4 ° 2θ ± 0.2 ° 2θ. In one aspect, the peaks at 18.7 ± 0.2 ° 2θ and 22.4 ° 2θ ± 0.2 ° 2θ have at least a medium or stronger relative intensity. In another aspect, the transmission X-ray powder diffraction pattern also comprises at least one, two, three or four peaks selected from the group consisting of 12.0 ± 0.2 ° 2θ, 18.4 ± 0.2 ° 2θ, 19.8 ± 0.2 ° 2θ and 21.7 ° 2θ ± 0.2 ° 2θ. In another aspect, the transmission X-ray powder diffraction pattern also comprises peaks at 12.0 ± 0.2 ° 2θ, 18.4 ± 0.2 ° 2θ, 19.8 ± 0.2 ° 2θ and 21.7 ° 2θ ± 0.2 ° 2θ. In another aspect, the transmission X-ray powder diffraction pattern further comprises at least one, two, three or four peaks selected from the group consisting of 13.1 ± 0.2 ° 2θ, 14.4 ± 0.2 ° 2θ, 17.5 ± 0.2 ° 2θ and 21.1 ° 2θ ± 0.2 ° 2θ. In another aspect, the transmission X-ray powder diffraction pattern further comprises peaks at 13.1 ± 0.2 ° 2θ, 14.4 ± 0.2 ° 2θ, 17.5 ± 0.2 ° 2θ and 21.1 ° 2θ ± 0.2 ° 2θ. In another aspect, the transmission X-ray powder diffraction pattern comprises at 8.9°2θ±0.2°2θ, 12.0°2θ±0.2°2θ, 13.1°2θ±0.2°2θ, 14.4°2θ±0.2°2θ, 17.5°2θ±0.2°2θ, 18.0°2θ±0.2°2θ, 18.4°2θ±0.2°2θ, 18.7°2θ±0.2°2θ, 19.8°2θ±0.2°2θ, 20.3°2θ±0.2°2θ, 20.6°2θ±0.2°2θ, In another aspect, the transmission X-ray powder diffraction pattern comprises peaks at 2θ±0.2°2θ, 21.1°2θ±0.2°2θ, 21.7°2θ±0.2°2θ, 22.4°2θ±0.2°2θ, 22.9°2θ±0.2°2θ, 25.2°2θ±0.2°2θ, 25.6°2θ±0.2°2θ, 26.2°2θ±0.2°2θ, 28.7°2θ±0.2°2θ, 30.4°2θ±0.2°2θ, 30.9°2θ±0.2°2θ and 31.1°2θ±0.2°2θ. In another aspect, the transmission X-ray powder diffraction pattern comprises peaks at 2θ±0.2°2θ, 21.1°2θ±0.2°2θ, 21.7°2θ±0.2°2θ, 22.4°2θ±0.2°2θ, 22.9°2θ±0.2°2θ, 25.2°2θ±0.2°2θ, 25.6°2θ±0.2°2θ, Fig. 9 The transmission X-ray powder diffraction patterns are essentially the same.

[0101] In some embodiments, Form B is characterized by: solid state 13C NMR spectrum comprising at least one peak selected from the group consisting of 166.9 ± 0.2 ppm, 130.2 ± 0.2 ppm, 118.6 ± 0.2 ppm, 117.4 ± 0.2 ppm, 110.0 ± 0.2 ppm, 49.9 ± 0.2 ppm, 46.6 ± 0.2 ppm, and 34.5 ± 0.2 ppm. In one aspect, the solid 13 The C NMR spectrum contains peaks at 49.9 ± 0.2 ppm and 46.6 ± 0.2 ppm. In another aspect, the solid 13 The C NMR spectrum comprises at least five peaks selected from the group consisting of 166.9 ± 0.2 ppm, 130.2 ± 0.2 ppm, 118.6 ± 0.2 ppm, 117.4 ± 0.2 ppm, 110.0 ± 0.2 ppm, 49.9 ± 0.2 ppm, 46.6 ± 0.2 ppm, and 34.5 ± 0.2 ppm. 13 The C NMR spectrum comprises peaks at 166.9±0.2 ppm, 118.6±0.2 ppm, 110.0±0.2 ppm, 49.9±0.2 ppm, and 46.6±0.2 ppm. In another aspect, the solid 13 The C NMR spectrum comprises peaks at 166.9±0.2 ppm, 130.2±0.2 ppm, 118.6±0.2 ppm, 117.4±0.2 ppm, 110.0±0.2 ppm, 49.9±0.2 ppm, 46.6±0.2 ppm, and 34.5±0.2 ppm. In another aspect, the solid 13 The C NMR spectrum comprises peaks at 167.7±0.2 ppm, 166.9±0.2 ppm, 147.3±0.2 ppm, 143.1±0.2 ppm, 139.9±0.2 ppm, 130.2±0.2 ppm, 125±0.2 ppm, 119.5±0.2 ppm, 118.6±0.2 ppm, 117.4±0.2 ppm, 110.0±0.2 ppm, 49.9±0.2 ppm, 46.6±0.2 ppm, and 34.5±0.2 ppm. In another aspect, the solid 13 The C NMR spectrum does not comprise at least one peak selected from the group consisting of 166.1 ± 0.2 ppm, 130.7 ± 0.2 ppm, 117.9 ± 0.2 ppm, 108.6 ± 0.2 ppm, and 35.2 ± 0.2 ppm. 13 The C NMR spectrum does not contain peaks at 166.1 ± 0.2 ppm, 117.9 ± 0.2 ppm, and 108.6 ± 0.2 ppm. In another aspect, the solid 13The C NMR spectrum did not contain peaks at 166.1±0.2 ppm, 130.7±0.2 ppm, 117.9±0.2 ppm, 108.6±0.2 ppm, and 35.2±0.2 ppm.

[0102] In some embodiments, Form B is characterized by a differential scanning calorimetry curve comprising a melting endotherm with an onset between about 185°C and about 200°C. In one aspect, the endotherm starts at about 185°C to about 197°C. In another aspect, the endotherm starts at 191°C ± 5°C. In another aspect, the differential scanning calorimetry curve is Fig.12 The differential scanning calorimetry curves of

[0103] In some embodiments, Form B is characterized by a thermogravimetric analysis thermogram, wherein the crystalline form exhibits a weight loss of less than about 0.5 wt % from about 25°C to about 110°C. In one aspect, the weight loss is less than about 0.2 wt %. In another aspect, the weight loss is less than about 0.1 wt %. In another aspect, the thermogravimetric analysis thermogram is characterized by a weight loss of less than about 0.5 wt % from about 25°C to about 110°C. Fig.13 The thermogravimetric analysis thermograms of the samples were essentially the same.

[0104] In some embodiments, Form B is characterized by a gravimetric vapor sorption curve, wherein the crystalline form exhibits a reversible moisture uptake of less than about 0.5 wt % from about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C. In one aspect, the reversible moisture uptake is less than about 0.2 wt %. In another aspect, the reversible moisture uptake is less than about 0.1 wt %. In another aspect, the gravimetric vapor sorption curve is less than about 0.5 wt %. Fig.14 The weight vapor adsorption curves are essentially the same.

[0105] In some embodiments, Form B is characterized by the physical characterization embodiments described above (transmission X-ray powder diffraction, solid state 13 The method can be characterized by at least two of C NMR, differential scanning calorimetry, thermogravimetric analysis, and / or gravimetric vapor sorption).

[0106] In some embodiments, Form B is characterized by the physical characterization embodiments described above (transmission X-ray powder diffraction, solid state 13 The method can be characterized by at least three of C NMR, differential scanning calorimetry, thermogravimetric analysis, and / or gravimetric vapor sorption).

[0107] In some embodiments, Form B is characterized by the physical characterization embodiments described above (transmission X-ray powder diffraction, solid state 13 The method can be characterized by at least four of C NMR, differential scanning calorimetry, thermogravimetric analysis, and / or gravimetric vapor sorption).

[0108] In some embodiments, Form B is characterized by:

[0109] a transmission X-ray powder diffraction pattern comprising peaks at 18.7 ± 0.2° 2θ and 22.4° 2θ ± 0.2° 2θ;

[0110] Solid State 13 C NMR spectrum comprising at least five peaks selected from the group consisting of 166.9±0.2 ppm, 130.2±0.2 ppm, 118.6±0.2 ppm, 117.4±0.2 ppm, 110.0±0.2 ppm, 49.9±0.2 ppm, 46.6±0.2 ppm, and 34.5±0.2 ppm; and

[0111] A differential scanning calorimetry curve comprising a melting endotherm with an onset between about 185°C and about 200°C.

[0112] In some embodiments, Form B is characterized by:

[0113] a transmission X-ray powder diffraction pattern comprising peaks at 18.7 ± 0.2° 2θ and 22.4° 2θ ± 0.2° 2θ;

[0114] Solid State 13 C NMR spectrum comprising peaks at 166.9±0.2 ppm, 118.6±0.2 ppm, 110.0±0.2 ppm, 49.9±0.2 ppm, and 46.6±0.2 ppm;

[0115] a differential scanning calorimetry curve comprising a melting endotherm with an onset between about 185°C and about 200°C; and

[0116] A thermogravimetric analysis thermogram wherein the crystalline form exhibits a weight loss of less than about 0.2 weight percent from about 25°C to about 110°C.

[0117] In some embodiments, Form B is characterized by:

[0118] a transmission X-ray powder diffraction pattern comprising peaks at 18.7 ± 0.2° 2θ and 22.4° 2θ ± 0.2° 2θ;

[0119] Solid State 13 C NMR spectrum comprising peaks at 166.9±0.2 ppm, 118.6±0.2 ppm, 110.0±0.2 ppm, 49.9±0.2 ppm, and 46.6±0.2 ppm;

[0120] a differential scanning calorimetry curve comprising a melting endotherm with an onset between about 185°C and about 200°C; and

[0121] A gravimetric vapor sorption plot wherein the crystalline form exhibits a reversible moisture uptake of less than about 0.2 wt % from about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C.

[0122] In some embodiments, Form B is characterized by:

[0123] a transmission X-ray powder diffraction pattern comprising peaks at 18.7 ± 0.2° 2θ and 22.4° 2θ ± 0.2° 2θ;

[0124] Solid State 13 C NMR spectrum comprising peaks at 166.9±0.2 ppm, 130.2±0.2 ppm, 118.6±0.2 ppm, 117.4±0.2 ppm, 110.0±0.2 ppm, 49.9±0.2 ppm, 46.6±0.2 ppm, and 34.5±0.2 ppm;

[0125] a differential scanning calorimetry curve comprising a melting endotherm with an onset at 191°C ± 5°C;

[0126] a thermogravimetric analysis thermogram, wherein the crystalline form exhibits a weight loss of less than about 0.1 wt % from about 25°C to about 110°C; and

[0127] A gravimetric vapor sorption plot wherein the crystalline form exhibits a reversible moisture uptake of less than about 0.1 wt % from about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C.

[0128] In some embodiments, Form B is a crystalline anhydrate.

[0129] In some embodiments, Form B has an elongated needle-like morphology.

[0130] In some embodiments, Form B is substantially free of any other crystalline forms of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0131] In some embodiments, Form B contains less than 5% by weight of any other crystalline forms of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0132] In some embodiments, Form B contains less than 10% by weight of any other crystalline forms of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0133] In some embodiments, the present disclosure provides a crystalline form of (R,S)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide (i.e., a crystalline form of racemic N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide).

[0134] In some embodiments, the present disclosure provides crystalline (R,S)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Type 2.

[0135] In some embodiments, Type 2 is characterized by a reflection X-ray powder diffraction pattern comprising at least one peak selected from the group consisting of 10.0±0.2°2θ, 12.9±0.2°2θ, 17.1±0.2°2θ, 22.0±0.2°2θ, and 22.8°2θ±0.2°2θ. In one aspect, the reflection X-ray powder diffraction pattern comprises peaks at 10.0±0.2°2θ, 12.9±0.2°2θ, 17.1±0.2°2θ, 22.0±0.2°2θ, and 22.8°2θ±0.2°2θ. In another aspect, the reflection X-ray powder diffraction pattern further comprises at least one, two, three, four or five peaks selected from the group consisting of 15.8 ± 0.2 ° 2θ, 16.2 ± 0.2 ° 2θ, 26.0 ± 0.2 ° 2θ, 26.5 ± 0.2 ° 2θ and 26.9 ° 2θ ± 0.2 ° 2θ. In another aspect, the reflection X-ray powder diffraction pattern further comprises peaks at 15.8 ± 0.2 ° 2θ, 16.2 ± 0.2 ° 2θ, 26.0 ± 0.2 ° 2θ, 26.5 ± 0.2 ° 2θ and 26.9 ° 2θ ± 0.2 ° 2θ. In another aspect, the reflection X-ray powder diffraction pattern further comprises peaks at 8.1 ± 0.2 ° 2θ, 10.0 ± 0.2 ° 2θ, 12.9 ± 0.2 ° 2θ, 15.6 ± 0.2 ° 2θ,

[0136] 15.8±0.2°2θ, 16.2±0.2°2θ, 17.1±0.2°2θ, 17.6±0.2°2θ, 17.9±0.2°2θ,

[0137] 19.2±0.2°2θ, 19.3±0.2°2θ, 20.7±0.2°2θ, 21.4±0.2°2θ, 22.0±0.2°2θ,

[0138] 22.8±0.2°2θ, 23.5±0.2°2θ, 24.0±0.2°2θ, 24.5±0.2°2θ, 26.1±0.2°2θ,

[0139] 26.5±0.2°2θ, 26.9±0.2°2θ, 29.5±0.2°2θ, 30.3±0.2°2θ, 30.7±0.2°2θ,

[0140] In another aspect, the reflection X-ray powder diffraction pattern is similar to Fig.15 The reflection X-ray powder diffraction patterns are essentially the same.

[0141] In some embodiments, Type 2 is characterized by a differential scanning calorimetry curve comprising a melting endotherm between about 195°C and about 210°C. In one aspect, the endotherm has an onset temperature between about 195°C and about 207°C. In another aspect, the endotherm has an onset temperature at 201°C ± 5°C. In another aspect, the differential scanning calorimetry curve is Fig.16 The differential scanning calorimetry curves of

[0142] In some embodiments, Type 2 is characterized by a gravimetric vapor sorption curve wherein the crystalline form exhibits a reversible moisture uptake of less than about 1.0 wt % from about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C. In one aspect, the reversible moisture uptake is less than about 0.7 wt %. In another aspect, the reversible moisture uptake is about 0.5 wt %. In another aspect, the gravimetric vapor sorption curve is Fig.17 The weight vapor adsorption curves are essentially the same.

[0143] In some embodiments, Type 2 is characterized by at least two of the above physical characterization embodiments (reflection X-ray powder diffraction, differential scanning calorimetry, and / or gravimetric vapor sorption).

[0144] In some embodiments, Type 2 is characterized by at least three of the above physical characterization embodiments (reflection X-ray powder diffraction, differential scanning calorimetry, and gravimetric vapor sorption).

[0145] In some embodiments, Type 2 is characterized by:

[0146] a reflection X-ray powder diffraction pattern comprising peaks at 10.0±0.2°2θ, 12.9±0.2°2θ, 17.1±0.2°2θ, 22.0±0.2°2θ, and 22.8°2θ±0.2°2θ;

[0147] a differential scanning calorimetry curve comprising a melting endotherm with an onset temperature between about 190°C and about 210°C; and

[0148] A gravimetric vapor sorption plot wherein the crystalline form exhibits a reversible moisture uptake of less than about 1.0 wt % from about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C.

[0149] In some embodiments, Type 2 is characterized by:

[0150] a reflection X-ray powder diffraction pattern comprising peaks at 10.0±0.2°2θ, 12.9±0.2°2θ, 17.1±0.2°2θ, 22.0±0.2°2θ, and 22.8°2θ±0.2°2θ;

[0151] a differential scanning calorimetry curve comprising a melting endotherm with an onset temperature between about 195°C and about 207°C; and

[0152] A gravimetric vapor sorption plot wherein the crystalline form exhibits a reversible moisture uptake of less than about 0.7 wt % from about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C.

[0153] In some embodiments, Type 2 is characterized by:

[0154] a reflection X-ray powder diffraction pattern comprising peaks at 10.0±0.2°2θ, 12.9±0.2°2θ, 17.1±0.2°2θ, 22.0±0.2°2θ, and 22.8°2θ±0.2°2θ;

[0155] a differential scanning calorimetry curve comprising a melting endotherm with an onset at 201°C ± 5°C; and

[0156] A gravimetric vapor sorption graph wherein the crystalline form exhibits a reversible moisture uptake of about 0.5 wt % from about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C.

[0157] In some embodiments, Type 2 is a crystalline anhydrate.

[0158] In some embodiments, Form 2 is substantially free of any other crystalline forms of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0159] In some embodiments, Type 2 contains less than 5% by weight of any other crystalline form of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0160] In some embodiments, Type 2 contains less than 10% by weight of any other crystalline form of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0161] In some embodiments, the present disclosure provides a composition comprising at least two crystalline forms selected from the group consisting of Form A, Form B, and Type 2.

[0162] In some embodiments, the present disclosure provides a composition comprising Form A and Form B. In one aspect, the composition comprises at least 50% by weight of Form B relative to Form A. In another aspect, the composition comprises at least 75% by weight of Form B relative to Form A. In another aspect, the composition comprises at least 90% by weight of Form B relative to Form A. In another aspect, the composition comprises at least 95% by weight of Form B relative to Form A. In another aspect, the composition comprises at least 95% by weight of Form B relative to any other crystalline form.

[0163] Representative Implementation Plans :

[0164] Embodiment 1: A crystalline form of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0165] Embodiment 2: According to the crystalline form of Embodiment 1, which is a crystalline form of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0166] Embodiment 3: The crystalline form of Embodiment 2, characterized by a transmission X-ray powder diffraction pattern comprising peaks at 18.7±0.2° 2θ and 22.4° 2θ±0.2° 2θ.

[0167] Embodiment 4: A crystalline form according to Embodiment 3, wherein the peaks at 18.7±0.2° 2θ and 22.4° 2θ±0.2° 2θ have at least moderate or stronger relative intensities.

[0168] Embodiment 5: The crystalline form of Embodiment 3, wherein the transmission X-ray powder diffraction pattern further comprises at least one peak selected from the group consisting of 12.0±0.2°2θ, 18.4±0.2°2θ, 19.8±0.2°2θ, and 21.7°2θ±0.2°2θ.

[0169] Embodiment 6: The crystalline form of Embodiment 3, wherein the transmission X-ray powder diffraction pattern further comprises peaks at 12.0±0.2°2θ, 18.4±0.2°2θ, 19.8±0.2°2θ, and 21.7°2θ±0.2°2θ.

[0170] Embodiment 7: The crystalline form of Embodiment 6, wherein the transmission X-ray powder diffraction pattern further comprises at least one peak selected from the group consisting of 13.1±0.2°2θ, 14.4±0.2°2θ, 17.5±0.2°2θ, and 21.1°2θ±0.2°2θ.

[0171] Embodiment 8: The crystalline form of Embodiment 6, wherein the transmission X-ray powder diffraction pattern further comprises peaks at 13.1±0.2°2θ, 14.4±0.2°2θ, 17.5±0.2°2θ, and 21.1°2θ±0.2°2θ.

[0172] Embodiment 9: The crystalline form of any one of Embodiments 3 to 8, wherein transmission x-ray powder diffraction is performed using Cu radiation.

[0173] Embodiment 10: The crystalline form according to any one of Embodiments 3 to 9, wherein transmission x-ray powder diffraction is performed using a PANalytical Empyrean diffractometer operating in transmission geometry, with a tube voltage of 45 kV and a filament emission of 40 mA.

[0174] Embodiment 11: A crystalline form according to any one of Embodiments 2 to 10, further characterized by: solid state 13 C NMR spectrum comprising at least one peak selected from the group consisting of 166.9±0.2 ppm, 130.2±0.2 ppm, 118.6±0.2 ppm, 117.4±0.2 ppm, 110.0±0.2 ppm, 49.9±0.2 ppm, 46.6±0.2 ppm, and 34.5±0.2 ppm.

[0175] Embodiment 12: A crystalline form according to embodiment 11, wherein the solid state 13 The C NMR spectrum contained peaks at 49.9±0.2 ppm and 46.6±0.2 ppm.

[0176] Embodiment 13: A crystalline form according to Embodiment 11, wherein the solid state 13 The C NMR spectrum comprises at least five peaks selected from the group consisting of 166.9±0.2 ppm, 130.2±0.2 ppm, 118.6±0.2 ppm, 117.4±0.2 ppm, 110.0±0.2 ppm, 49.9±0.2 ppm, 46.6±0.2 ppm, and 34.5±0.2 ppm.

[0177] Embodiment 14: A crystalline form according to embodiment 11, wherein the solid state 13The C NMR spectrum contained peaks at 166.9±0.2 ppm, 118.6±0.2 ppm, 110.0±0.2 ppm, 49.9±0.2 ppm, and 46.6±0.2 ppm.

[0178] Embodiment 15: A crystalline form according to Embodiment 11, wherein the solid state 13 The C NMR spectrum contained peaks at 166.9±0.2 ppm, 130.2±0.2 ppm, 118.6±0.2 ppm, 117.4±0.2 ppm, 110.0±0.2 ppm, 49.9±0.2 ppm, 46.6±0.2 ppm, and 34.5±0.2 ppm.

[0179] Embodiment 16: A crystalline form according to Embodiment 11, wherein the solid state 13 The C NMR spectrum contained peaks at 167.7±0.2 ppm, 166.9±0.2 ppm, 147.3±0.2 ppm, 143.1±0.2 ppm, 139.9±0.2 ppm, 130.2±0.2 ppm, 125±0.2 ppm, 119.5±0.2 ppm, 118.6±0.2 ppm, 117.4±0.2 ppm, 110.0±0.2 ppm, 49.9±0.2 ppm, 46.6±0.2 ppm, and 34.5±0.2 ppm.

[0180] Embodiment 17: A crystalline form according to any one of Embodiments 11 to 16, wherein the solid state 13 The C NMR spectrum does not include at least one peak selected from the group consisting of 166.1±0.2 ppm, 130.7±0.2 ppm, 117.9±0.2 ppm, 108.6±0.2 ppm, and 35.2±0.2 ppm.

[0181] Embodiment 18: A crystalline form according to any one of Embodiments 11 to 16, wherein the solid state 13 The C NMR spectrum did not contain peaks at 166.1±0.2 ppm, 117.9±0.2 ppm, and 108.6±0.2 ppm.

[0182] Embodiment 19: A crystalline form according to any one of Embodiments 11 to 16, wherein the solid state 13 The C NMR spectrum did not contain peaks at 166.1±0.2 ppm, 130.7±0.2 ppm, 117.9±0.2 ppm, 108.6±0.2 ppm, and 35.2±0.2 ppm.

[0183] Embodiment 20: The crystalline form of any one of Embodiments 2 to 19, further characterized by a differential scanning calorimetry curve comprising a melting endotherm with an onset between about 185°C and about 200°C.

[0184] Embodiment 21: The crystalline form of Embodiment 20, wherein the endotherm has an onset between about 185°C and about 197°C.

[0185] Embodiment 22: The crystalline form of Embodiment 20, wherein the endotherm has an onset at 191°C ± 5°C.

[0186] Embodiment 23: The crystalline form of Embodiment 20, wherein the differential scanning calorimetry curve is Fig.12 The differential scanning calorimetry curves of

[0187] Embodiment 24: A crystalline form according to any one of Embodiments 20 to 23, wherein differential scanning calorimetry is performed on a TA Instruments Differential Scanning Calorimeter (Model Q2000), wherein the sample is placed in an aluminum pan and heated at a rate of 10°C / minute to a temperature of 230°C under nitrogen.

[0188] Embodiment 25: The crystalline form of any one of Embodiments 3 to 24, further characterized by a thermogravimetric analysis thermogram, wherein the crystalline form exhibits a weight loss of less than about 0.5 wt % from about 25°C to about 110°C.

[0189] Embodiment 26: The crystalline form of Embodiment 25, wherein the weight loss is less than about 0.2 wt%.

[0190] Embodiment 27: The crystalline form of Embodiment 25, wherein the weight loss is less than about 0.1 wt%.

[0191] Embodiment 28: The crystalline form of Embodiment 25, wherein the thermogravimetric analysis thermogram is Fig.13 The thermogravimetric analysis thermograms of the samples were essentially the same.

[0192] Embodiment 29: The crystalline form of any one of Embodiments 3 to 28, further characterized by a gravimetric vapor sorption plot, wherein the crystalline form exhibits a reversible moisture uptake of less than about 0.5 wt % from about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C.

[0193] Embodiment 30: The crystalline form of Embodiment 29, wherein the reversible moisture uptake is less than about 0.2 wt%.

[0194] Embodiment 31: The crystalline form of Embodiment 29, wherein the reversible moisture uptake is less than about 0.1 wt%.

[0195] Embodiment 32: A crystalline form according to Embodiment 29, wherein the weight vapor sorption curve is Fig.14 The weight vapor adsorption curves are essentially the same.

[0196] Embodiment 33: The crystalline form according to any one of Embodiments 3 to 10, wherein the crystalline form is further characterized by:

[0197] Solid State 13 C NMR spectrum comprising at least five peaks selected from the group consisting of 166.9±0.2 ppm, 130.2±0.2 ppm, 118.6±0.2 ppm, 117.4±0.2 ppm, 110.0±0.2 ppm, 49.9±0.2 ppm, 46.6±0.2 ppm, and 34.5±0.2 ppm; and

[0198] A differential scanning calorimetry curve comprising a melting endotherm with an onset between about 185°C and about 200°C.

[0199] Embodiment 34: The crystalline form according to any one of Embodiments 3 to 10, wherein the crystalline form is further characterized by:

[0200] Solid State 13 C NMR spectrum comprising peaks at 166.9±0.2 ppm, 118.6±0.2 ppm, 110.0±0.2 ppm, 49.9±0.2 ppm, and 46.6±0.2 ppm;

[0201] A differential scanning calorimetry curve comprising a melting point onset between about 185°C and about 200°C.

[0202] heat of fusion; and

[0203] A thermogravimetric analysis thermogram wherein the crystalline form exhibits a weight loss of less than about 0.2 weight percent from about 25°C to about 110°C.

[0204] Embodiment 35: The crystalline form according to any one of Embodiments 3 to 10, wherein the crystalline form is further characterized by:

[0205] Solid State 13 C NMR spectrum comprising peaks at 166.9±0.2 ppm, 118.6±0.2 ppm, 110.0±0.2 ppm, 49.9±0.2 ppm, and 46.6±0.2 ppm;

[0206] A differential scanning calorimetry curve comprising a melting point onset between about 185°C and about 200°C.

[0207] heat of fusion; and

[0208] A gravimetric vapor sorption plot wherein the crystalline form exhibits a reversible moisture uptake of less than about 0.2 wt % from about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C.

[0209] Embodiment 36: The crystalline form according to any one of Embodiments 3 to 10, wherein the crystalline form is further characterized by:

[0210] Solid State 13 C NMR spectrum comprising peaks at 166.9±0.2 ppm, 130.2±0.2 ppm, 118.6±0.2 ppm, 117.4±0.2 ppm, 110.0±0.2 ppm, 49.9±0.2 ppm, 46.6±0.2 ppm, and 34.5±0.2 ppm;

[0211] a differential scanning calorimetry curve comprising a melting endotherm with an onset at 191°C ± 5°C;

[0212] Thermogravimetric analysis thermogram wherein the crystalline form exhibits less than

[0213] A weight loss of about 0.1 wt %; and

[0214] A gravimetric vapor sorption plot wherein the crystalline form exhibits a reversible moisture uptake of less than about 0.1 wt % from about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C.

[0215] Embodiment 37: The crystalline form according to any one of Embodiments 3 to 36, wherein the crystalline form is a crystalline anhydrate.

[0216] Embodiment 38: A crystalline form according to any one of Embodiments 3 to 37, wherein the crystalline form comprises less than 5 weight % of any other crystalline form of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0217] Embodiment 39: A crystalline form according to any one of Embodiments 3 to 37, wherein the crystalline form is substantially free of any other crystalline form of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0218] Embodiment 40: The crystalline form of Embodiment 2, wherein:

[0219] a transmission X-ray powder diffraction pattern comprising at least one peak selected from the group consisting of 12.0±0.2°2θ, 18.4±0.2°2θ, 19.8±0.2°2θ, and 21.7°2θ±0.2°2θ; and

[0220] The transmission X-ray powder diffraction pattern does not contain the 18.7 ± 0.2 ° 2θ and 22.4 ° 2θ ± 0.2 ° 2θ

[0221] Peaks with medium or stronger relative intensity.

[0222] Embodiment 41: The crystalline form of Embodiment 40, wherein the transmission X-ray powder diffraction pattern does not comprise peaks at 18.7±0.2° 2θ and 22.4° 2θ±0.2° 2θ.

[0223] Embodiment 42: The crystalline form of Embodiment 40, wherein the transmission X-ray powder diffraction pattern further comprises at least two peaks selected from the group consisting of 12.0±0.2°2θ, 18.4±0.2°2θ, 19.8±0.2°2θ, and 21.7°2θ±0.2°2θ.

[0224] Embodiment 43: The crystalline form of Embodiment 40, wherein the transmission X-ray powder diffraction pattern further comprises peaks at 12.0±0.2°2θ, 18.4±0.2°2θ, 19.8±0.2°2θ, and 21.7°2θ±0.2°2θ.

[0225] Embodiment 44: The crystalline form of Embodiment 43, wherein the transmission X-ray powder diffraction pattern further comprises at least one peak selected from the group consisting of 13.1±0.2°2θ, 14.4±0.2°2θ, 17.5±0.2°2θ, and 21.1°2θ±0.2°2θ.

[0226] Embodiment 45: The crystalline form of Embodiment 43, wherein the transmission X-ray powder diffraction pattern further comprises peaks at 13.1±0.2°2θ, 14.4±0.2°2θ, 17.5±0.2°2θ, and 21.1°2θ±0.2°2θ.

[0227] Embodiment 46: The crystalline form of any one of Embodiments 40 to 45, wherein transmission x-ray powder diffraction is performed using Cu radiation.

[0228] Embodiment 47: The crystalline form of any one of Embodiments 40 to 46, wherein transmission x-ray powder diffraction is performed using a PANalytical Empyrean diffractometer operating in transmission geometry, with a tube voltage of 45 kV and a filament emission of 40 mA.

[0229] Embodiment 48: A crystalline form according to any one of Embodiments 2 or 40 to 47, further characterized by: solid 13 C NMR spectrum comprising at least one peak selected from the group consisting of 166.1±0.2 ppm, 130.7±0.2 ppm, 117.9±0.2 ppm, 108.6±0.2 ppm, and 35.2±0.2 ppm.

[0230] Embodiment 49: A crystalline form according to Embodiment 48, wherein the solid state 13 The C NMR spectrum contained peaks at 166.1±0.2 ppm, 117.9±0.2 ppm, and 108.6±0.2 ppm.

[0231] Embodiment 50: A crystalline form according to Embodiment 48, wherein the solid state 13 The C NMR spectrum contained peaks at 166.1±0.2 ppm, 130.7±0.2 ppm, 117.9±0.2 ppm, 108.6±0.2 ppm, and 35.2±0.2 ppm.

[0232] Embodiment 51: A crystalline form according to Embodiment 48, wherein the solid state 13 The C NMR spectrum contained peaks at 168.0±0.2 ppm, 166.1±0.2 ppm, 147.5±0.2 ppm, 146.4±0.2 ppm, 143.1±0.2 ppm, 139.5±0.2 ppm, 130.7±0.2 ppm, 125±0.2 ppm, 117.9±0.2 ppm, 108.6±0.2 ppm, 67.1±0.2 ppm, 48.0±0.2 ppm, 42.9±0.2 ppm, and 35.2±0.2 ppm.

[0233] Embodiment 52: A crystalline form according to any one of Embodiments 48 to 51, wherein the solid state 13 The C NMR spectrum does not include at least one peak selected from the group consisting of 166.9±0.2 ppm, 130.2±0.2 ppm, 118.6±0.2 ppm, 117.4±0.2 ppm, 110.0±0.2 ppm, 49.9±0.2 ppm, 46.6±0.2 ppm, and 34.5±0.2 ppm.

[0234] Embodiment 53: A crystalline form according to any one of Embodiments 48 to 51, wherein the solid state 13The C NMR spectrum does not include at least five peaks selected from the group consisting of 166.9±0.2 ppm, 130.2±0.2 ppm, 118.6±0.2 ppm, 117.4±0.2 ppm, 110.0±0.2 ppm, 49.9±0.2 ppm, 46.6±0.2 ppm, and 34.5±0.2 ppm.

[0235] Embodiment 54: A crystalline form according to any one of Embodiments 48 to 51, wherein the solid state 13 The C NMR spectrum did not contain peaks at 49.9±0.2 ppm and 46.6±0.2 ppm.

[0236] Embodiment 55: A crystalline form according to any one of Embodiments 48 to 51, wherein the solid state 13 The C NMR spectrum did not contain peaks at 166.9±0.2 ppm, 118.6±0.2 ppm, 110.0±0.2 ppm, 49.9±0.2 ppm, and 46.6±0.2 ppm.

[0237] Embodiment 56: A crystalline form according to any one of Embodiments 48 to 51, wherein the solid state 13 The C NMR spectrum does not include peaks at 166.9±0.2 ppm, 130.2±0.2 ppm, 118.6±0.2 ppm, 117.4±0.2 ppm, 110.0±0.2 ppm, 49.9±0.2 ppm, 46.6±0.2 ppm, and 34.5±0.2 ppm.

[0238] Embodiment 57: The crystalline form of any one of Embodiment 2 or Embodiments 40 to 56, further characterized by a differential scanning calorimetry curve comprising a melting endotherm with an onset temperature between about 165°C and about 180°C.

[0239] Embodiment 58: The crystalline form of Embodiment 57, wherein the endotherm comprises a melting endotherm with an onset temperature between about 165°C and about 177°C.

[0240] Embodiment 59: The crystalline form of Embodiment 57, wherein the endotherm has an onset at 171 °C ± 5 °C.

[0241] Embodiment 60: The crystalline form of Embodiment 57, wherein the differential scanning calorimetry curve is Figure 6 The differential scanning calorimetry curves of

[0242] Embodiment 61: The crystalline form according to any one of Embodiments 57 to 60, wherein differential scanning calorimetry is performed on a TA Instruments Differential Scanning Calorimeter (Model Q2000), wherein the sample is placed in an aluminum pan and heated at a rate of 10°C / minute to a temperature of 230°C under nitrogen.

[0243] Embodiment 62: The crystalline form of any one of Embodiments 40 to 61, further characterized by a thermogravimetric analysis thermogram, wherein the crystalline form exhibits a weight loss of less than about 0.5 wt% from about 25°C to about 110°C.

[0244] Embodiment 63: The crystalline form of Embodiment 62, wherein the weight loss is less than about 0.2 wt%.

[0245] Embodiment 64: The crystalline form of Embodiment 62, wherein the weight loss is less than about 0.1 wt%.

[0246] Embodiment 65: The crystalline form of Embodiment 62, wherein the thermogravimetric analysis thermogram is Figure 7 The thermogravimetric analysis thermograms of the samples were essentially the same.

[0247] Embodiment 66: The crystalline form of any one of Embodiments 40 to 65, further characterized by a gravimetric vapor sorption plot, wherein the crystalline form exhibits a reversible moisture uptake of less than about 0.5 wt % from about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C.

[0248] Embodiment 67: The crystalline form of Embodiment 66, wherein the reversible moisture uptake is less than about 0.2 wt%.

[0249] Embodiment 68: The crystalline form of Embodiment 66, wherein the reversible moisture uptake is less than about 0.1 wt%.

[0250] Embodiment 69: The crystalline form of Embodiment 66, wherein the weight vapor sorption curve is Figure 8 The weight vapor adsorption curves are essentially the same.

[0251] Embodiment 70: The crystalline form according to any one of Embodiments 40 to 47, wherein the crystalline form is further characterized by:

[0252] Solid State 13 C NMR spectrum comprising peaks at 166.1±0.2 ppm, 130.7±0.2 ppm, 117.9±0.2 ppm, 108.6±0.2 ppm, and 35.2±0.2 ppm; and

[0253] A differential scanning calorimetry curve comprising a melting endotherm with an onset temperature between about 165°C and about 180°C.

[0254] Embodiment 71: The crystalline form according to any one of Embodiments 40 to 47, wherein the crystalline form is further characterized by:

[0255] Solid State 13 C NMR spectrum comprising peaks at 166.1±0.2 ppm, 130.7±0.2 ppm, 117.9±0.2 ppm, 108.6±0.2 ppm, and 35.2±0.2 ppm;

[0256] a differential scanning calorimetry curve comprising a melting endotherm with an onset temperature between about 165°C and about 177°C; and

[0257] A thermogravimetric analysis thermogram wherein the crystalline form exhibits a weight loss of less than about 0.2 weight percent from about 25°C to about 110°C.

[0258] Embodiment 72: The crystalline form according to any one of Embodiments 40 to 47, wherein the crystalline form is further characterized by:

[0259] Solid State 13 C NMR spectrum comprising peaks at 166.1±0.2 ppm, 130.7±0.2 ppm, 117.9±0.2 ppm, 108.6±0.2 ppm, and 35.2±0.2 ppm;

[0260] a differential scanning calorimetry curve comprising a melting endotherm with an onset temperature between about 165°C and about 177°C; and

[0261] A gravimetric vapor sorption plot wherein the crystalline form exhibits a reversible moisture uptake of less than about 0.2 wt % from about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C.

[0262] Embodiment 73: The crystalline form according to any one of Embodiments 40 to 47, wherein the crystalline form is further characterized by:

[0263] Solid State 13 C NMR spectrum, which contains 166.1±0.2ppm, 130.7±0.2ppm,

[0264] peaks at 117.9 ± 0.2 ppm, 108.6 ± 0.2 ppm, and 35.2 ± 0.2 ppm;

[0265] a differential scanning calorimetry curve comprising a melting endotherm with an onset at 171°C ± 5°C;

[0266] Thermogravimetric analysis thermogram wherein the crystalline form exhibits less than

[0267] A weight loss of about 0.2 wt. %; and

[0268] A gravimetric vapor sorption plot wherein the crystalline form exhibits a reversible moisture uptake of less than about 0.2 wt % from about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C.

[0269] Embodiment 74: The crystalline form according to any one of Embodiments 40 to 73, wherein the crystalline form is a crystalline anhydrate.

[0270] Embodiment 75: A crystalline form according to any one of Embodiments 40 to 74, wherein the crystalline form comprises less than 5 weight % of any other crystalline form of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0271] Embodiment 76: A crystalline form according to any one of Embodiments 40 to 74, wherein the crystalline form is substantially free of any other crystalline form of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0272] Embodiment 77: According to the crystalline form of Embodiment 1, which is a crystalline form of (R,S)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0273] Embodiment 78: The crystalline form of Embodiment 77, characterized by a reflection X-ray powder diffraction pattern comprising at least one peak selected from the group consisting of 10.0±0.2°2θ, 12.9±0.2°2θ, 17.1±0.2°2θ, 22.0±0.2°2θ, and 22.8°2θ±0.2°2θ.

[0274] Embodiment 79: The crystalline form of Embodiment 78, wherein the reflection X-ray powder diffraction pattern comprises peaks at 10.0±0.2°2θ, 12.9±0.2°2θ, 17.1±0.2°2θ, 22.0±0.2°2θ, and 22.8°2θ±0.2°2θ.

[0275] Embodiment 80: The crystalline form of Embodiment 79, wherein the reflection X-ray powder diffraction pattern further comprises at least one peak selected from the group consisting of 15.8±0.2°2θ, 16.2±0.2°2θ, 26.0±0.2°2θ, 26.5±0.2°2θ, and 26.9°2θ±0.2°2θ.

[0276] Embodiment 81: The crystalline form of Embodiment 79, wherein the reflection X-ray powder diffraction pattern further comprises peaks at 15.8±0.2°2θ, 16.2±0.2°2θ, 26.0±0.2°2θ, 26.5±0.2°2θ, and 26.9°2θ±0.2°2θ.

[0277] Embodiment 82: The crystalline form of any one of Embodiments 77 to 81, wherein reflection x-ray powder diffraction is performed using Cu radiation.

[0278] Embodiment 83: The crystalline form of any one of Embodiments 77 to 82, wherein reflection x-ray powder diffraction is performed using a PANalytical Empyrean diffractometer operating in reflection geometry, with a tube voltage of 45 kV and a filament emission of 40 mA.

[0279] Embodiment 84: The crystalline form of any one of Embodiments 77 to 83, further characterized by a differential scanning calorimetry curve comprising a melting endotherm between about 195°C and about 210°C.

[0280] Embodiment 85: The crystalline form of Embodiment 84, wherein the endotherm has an onset temperature between about 195°C and about 207°C.

[0281] Embodiment 86: The crystalline form of Embodiment 84, wherein the endotherm has an onset temperature at 201 °C ± 5 °C.

[0282] Embodiment 87: The crystalline form of Embodiment 85, wherein the differential scanning calorimetry curve is Fig.16 The differential scanning calorimetry curves of

[0283] Embodiment 88: A crystalline form according to any one of Embodiments 84 to 87, wherein differential scanning calorimetry is performed on a TA Instruments Differential Scanning Calorimeter (Model Q2000), wherein the sample is placed in an aluminum pan and heated at a rate of 10°C / minute to a temperature of 230°C under nitrogen.

[0284] Embodiment 89: The crystalline form of any one of Embodiments 77 to 88, further characterized by a gravimetric vapor sorption plot, wherein the crystalline form exhibits a reversible moisture uptake of less than about 1.0 wt % from about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C.

[0285] Embodiment 90: The crystalline form of Embodiment 89, wherein the reversible moisture uptake is less than about 0.7 wt%.

[0286] Embodiment 91: The crystalline form of Embodiment 89, wherein the reversible moisture uptake is about 0.5 wt%.

[0287] Embodiment 92: The crystalline form of Embodiment 89, wherein the weight vapor sorption curve is Fig.17 The weight vapor adsorption curves are essentially the same.

[0288] Embodiment 93: The crystalline form according to any one of Embodiments 78 to 92, wherein the crystalline form is further characterized by:

[0289] a differential scanning calorimetry curve comprising a melting endotherm with an onset temperature between about 190°C and about 210°C; and

[0290] A gravimetric vapor sorption plot wherein the crystalline form exhibits a reversible moisture uptake of less than about 1.0 wt % from about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C.

[0291] Embodiment 94: The crystalline form according to any one of Embodiments 78 to 92, wherein the crystalline form is further characterized by:

[0292] a differential scanning calorimetry curve comprising a melting endotherm with an onset temperature between about 195°C and about 207°C; and

[0293] A gravimetric vapor sorption plot wherein the crystalline form exhibits a reversible moisture uptake of less than about 0.7 wt % from about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C.

[0294] Embodiment 95: The crystalline form according to any one of Embodiments 78 to 92, wherein the crystalline form is further characterized by:

[0295] a differential scanning calorimetry curve comprising a melting endotherm with an onset at 201°C ± 5°C;

[0296] as well as

[0297] A gravimetric vapor sorption graph wherein the crystalline form exhibits a reversible moisture uptake of about 0.5 wt % from about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C.

[0298] Embodiment 96: The crystalline form according to any one of Embodiments 78 to 95, wherein the crystalline form is a crystalline anhydrate.

[0299] Embodiment 97: A crystalline form according to any one of Embodiments 78 to 96, wherein the crystalline form comprises less than 5% by weight of any other crystalline form of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0300] Embodiment 98: The crystalline form of any one of Embodiments 78 to 96, wherein the crystalline form is substantially free of any other crystalline form of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0301] Embodiment 99: A composition comprising at least two crystalline forms selected from the group consisting of:

[0302] The crystalline form according to any one of embodiments 3 to 37;

[0303] The crystalline form according to any one of embodiments 40 to 74; and

[0304] A crystalline form according to any one of embodiments 77 to 96.

[0305] Embodiment 100: The composition according to embodiment 99, wherein the composition comprises:

[0306] The crystalline form according to any one of embodiments 3 to 37; and

[0307] The crystalline form according to any one of embodiments 40 to 74.

[0308] Embodiment 101: The composition of embodiment 100, wherein the composition comprises at least 50% by weight of the crystalline form according to any one of embodiments 3 to 37 relative to the crystalline form according to any one of embodiments 40 to 74.

[0309] Embodiment 102: The composition of embodiment 100, wherein the composition comprises at least 75% by weight of the crystalline form according to any one of embodiments 3 to 37 relative to the crystalline form according to any one of embodiments 40 to 74.

[0310] Embodiment 103: The composition of embodiment 100, wherein the composition comprises at least 90% by weight of the crystalline form according to any one of embodiments 3 to 37 relative to the crystalline form according to any one of embodiments 40 to 74.

[0311] Embodiment 104: The composition of embodiment 100, wherein the composition comprises at least 95% by weight of the crystalline form according to any one of embodiments 3 to 37 relative to the crystalline form according to any one of embodiments 40 to 74.

[0312] Embodiment 105: The composition of Embodiment 100, wherein the composition comprises at least 95% by weight of the crystalline form according to any one of Embodiments 3 to 37 relative to any other crystalline form.

[0313] Embodiment 106: A pharmaceutical composition comprising the crystalline form according to any one of Embodiments 1 to 98 and one or more pharmaceutically acceptable excipients.

[0314] Embodiment 107: The pharmaceutical composition according to Embodiment 106, wherein the pharmaceutical composition comprises the crystalline form according to any one of Embodiments 3 to 37.

[0315] Embodiment 108: The pharmaceutical composition according to Embodiment 107, wherein the pharmaceutical composition further comprises the crystalline form according to any one of Embodiments 40 to 74.

[0316] Embodiment 109: The pharmaceutical composition according to Embodiment 106, wherein the pharmaceutical composition comprises the crystalline form according to any one of Embodiments 40 to 70.

[0317] Embodiment 110: The pharmaceutical composition according to any one of Embodiments 106 to 109, wherein the pharmaceutical composition is a solid pharmaceutical composition.

[0318] Embodiment 111: A method of treating or preventing a FAP-mediated disorder in a subject suffering from or susceptible to a FAP-mediated disorder, the method comprising administering to the subject a therapeutically effective amount of a crystalline form according to any one of Embodiments 1 to 98.

[0319] Embodiment 112: The method according to embodiment 111, wherein the FAP-mediated disorder is selected from the group consisting of liver disease, type 2 diabetes, cardiovascular disorders, obesity, obesity-related disorders, fibrosis, keloid disorders, inflammation and cancer.

[0320] Embodiment 113: The method of Embodiment 112, wherein the FAP-mediated disorder is liver disease.

[0321] Embodiment 114: The method of Embodiment 113, wherein the liver disease is non-alcoholic steatohepatitis.

[0322] Embodiment 115: Use of a compound according to any one of Embodiments 1 to 98, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating or preventing a FAP-mediated disorder.

[0323] III. How to use

[0324] (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide is an inhibitor of prolyl endopeptidase fibroblast activation protein (FAP) activity. FAP is an endopeptidase that enzymatically cleaves substrates involved in glucose and lipid metabolism, fibrinolysis, and collagen production.

[0325] FAP is believed to cleave and inactivate human fibroblast growth factor 21 (FGF-21) (Biochem. J. 2016, 473, 605), a protein involved in regulating glucose and lipid metabolism. It is hypothesized that inhibition of FAP increases endogenous FGF-21 levels and signaling, and leads to, for example, reduced steatosis, improved insulin sensitivity, improved glucose tolerance, reduced body weight, and / or reduced cardiovascular mortality.

[0326] FAP is also believed to cut human α2-antiplasmin (α2AP) (Blood 2004 103, 3783), which is a protein involved in regulating fibrosis and fibrinolysis. Tissue repair involves coagulation, resulting in fibrin deposition. When converted from its inactive form (plasminogen) by plasminogen activators, the fibrin of the clot is usually mainly cleaved by plasmin. Fibrinolysis is inhibited by plasminogen activator inhibitor-1 (PAI-1), plasminogen activator inhibitor-2 (PAI-2) and α2AP (Experimental & Molecular Medicine 2020, 52, 367), all of which are induced by tissue trauma. FAP converts α2AP into a form that binds to fibrin more effectively, which reduces the plasmin degradation of fibrin at the site of injury. It is hypothesized that inhibition of FAP increases fibrinolysis and improves tissue regeneration at the site of injury (J. Thromb. Haemost. 2013, 11, 2029; Proteomics Clin. Appl. 2014, 8, 454).

[0327] FAP is also believed to promote collagen production and deposition and play a role in increased fibrosis by altering extracellular matrix (ECM) turnover (J Biol Chem 2016, 8, 291). It is hypothesized that inhibition of FAP leads to a decrease in collagen deposition and a decrease in inflammation (Inflamm. Bowel Dis. 2018, 18, 332).

[0328] In view of the above, it is hypothesized that inhibition of FAP would jointly reduce fibrosis and inflammation by decreasing hepatic stellate cell activity and increasing fibrinolysis, and further provide positive metabolic effects by increasing FGF21 signaling and improving glucose tolerance.

[0329] Thus, in some embodiments, the present disclosure provides a method for treating or preventing a FAP-mediated disorder in a subject in need thereof by administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure.

[0330] In some embodiments, the present disclosure provides a method for treating or preventing a disorder characterized by overexpression of FAP in a subject in need thereof by administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure.

[0331] In some embodiments, the present disclosure provides a method for treating or preventing a liver disease in a subject by administering a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure to a subject in need thereof. In one aspect, the liver disease is fatty liver disease. In another aspect, the liver disease is non-alcoholic fatty liver disease (NAFLD). In another aspect, NAFLD is selected from the group consisting of isolated steatosis, non-alcoholic steatohepatitis (NASH), liver fibrosis and cirrhosis. In another aspect, the liver disease is end-stage liver disease. In another aspect, the subject also suffers from or is susceptible to one or more diseases selected from the group consisting of obesity, dyslipidemia, insulin resistance, type 2 diabetes and renal insufficiency.

[0332] In some embodiments, the present disclosure provides a method for treating liver disease in a subject in need thereof by administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure, wherein the subject has a liver disease of 27 kg / m 2 Up to 40kg / m 2 In one aspect, the subject has a body mass index (BMI) of 30 kg / m 2 Up to 39.9kg / m 2 In another aspect, the subject has a BMI of at least 40 kg / m 2 In another aspect, the subject is overweight. In another aspect, the subject is obese. In another aspect, the liver disease is NAFLD. In another aspect, the liver disease is NASH. In another aspect, the liver disease is liver fibrosis. In another aspect, the liver disease is cirrhosis.

[0333] In some embodiments, the present disclosure provides a method for treating a subject's liver disease by administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure, wherein the subject also suffers from or is susceptible to dyslipidemia. In another aspect, the liver disease is NAFLD. In another aspect, the liver disease is NASH. In another aspect, the liver disease is liver fibrosis. In another aspect, the liver disease is cirrhosis.

[0334] In some embodiments, the present disclosure provides a method for treating a subject's liver disease by administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure, wherein the subject also suffers from or is susceptible to insulin resistance. In another aspect, the liver disease is NAFLD. In another aspect, the liver disease is NASH. In another aspect, the liver disease is liver fibrosis. In another aspect, the liver disease is cirrhosis.

[0335] In some embodiments, the present disclosure provides a method for treating a subject's liver disease by administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure, wherein the subject also suffers from or is susceptible to at least one of type 2 diabetes and renal insufficiency. In another aspect, the liver disease is NAFLD. In another aspect, the liver disease is NASH. In another aspect, the liver disease is liver fibrosis. In another aspect, the liver disease is cirrhosis.

[0336] In some embodiments, the present disclosure provides a method for treating a subject's liver disease by administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure, wherein the subject also suffers from or is susceptible to type 2 diabetes. In another aspect, the liver disease is NAFLD. In another aspect, the liver disease is NASH. In another aspect, the liver disease is liver fibrosis. In another aspect, the liver disease is cirrhosis.

[0337] In some embodiments, the present disclosure provides a method for treating a subject's liver disease by administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure, wherein the subject also suffers from or is susceptible to renal insufficiency. In another aspect, the liver disease is NAFLD. In another aspect, the liver disease is NASH. In another aspect, the liver disease is liver fibrosis. In another aspect, the liver disease is cirrhosis.

[0338] In some embodiments, the present disclosure provides a method for reducing liver fat in a subject by administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure. In one aspect, the subject suffers from or is susceptible to NAFLD. In another aspect, the subject suffers from or is susceptible to NASH. In another aspect, the subject suffers from or is susceptible to liver fibrosis. In another aspect, the subject suffers from or is susceptible to cirrhosis. In another aspect, the subject also suffers from or is susceptible to one or more conditions selected from the group consisting of obesity, dyslipidemia, insulin resistance, type 2 diabetes, and renal insufficiency.

[0339] In some embodiments, the present disclosure provides a method for treating or preventing nonalcoholic fatty liver disease (NAFLD) in a subject by administering to the subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure. In one aspect, NAFLD is stage 1 NAFLD. In another aspect, NAFLD is stage 2 NAFLD. In another aspect, NAFLD is stage 3 NAFLD. In another aspect, NAFLD is stage 4 NAFLD. See, e.g., "The Diagnosis and Management of Nonalcoholic Fatty Liver Disease: Practice Guidance From the American Association for the Study of Liver Diseases," Hepatology, 2018, Vol. 67, No. 1. In another aspect, the subject also suffers from or is susceptible to one or more conditions selected from the group consisting of obesity, dyslipidemia, insulin resistance, type 2 diabetes, and renal insufficiency.

[0340] In some embodiments, the present disclosure provides a method for treating or preventing nonalcoholic steatohepatitis (NASH) in a subject by administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure. In one aspect, the NASH is stage 1 NASH. In another aspect, the NASH is stage 2 NASH. In another aspect, the NASH is stage 3 NASH. In another aspect, the NASH is stage 4 NASH. In another aspect, the subject also suffers from or is susceptible to one or more conditions selected from the group consisting of obesity, dyslipidemia, insulin resistance, type 2 diabetes, and renal insufficiency.

[0341] In some embodiments, the present disclosure provides a method for treating or preventing liver fibrosis in a subject by administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure. In one aspect, the subject suffers from stage 3 liver fibrosis. In another aspect, the subject also suffers from or is susceptible to one or more conditions selected from the group consisting of obesity, dyslipidemia, insulin resistance, type 2 diabetes, and renal insufficiency.

[0342] In some embodiments, the present disclosure provides a method for treating or preventing cirrhosis of the subject by administering to the subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure. In one aspect, the subject suffers from stage F4 cirrhosis. In another aspect, the subject also suffers from or is susceptible to one or more conditions selected from the group consisting of obesity, dyslipidemia, insulin resistance, type 2 diabetes, and renal insufficiency.

[0343] In some embodiments, the present disclosure provides a method for treating or preventing type 2 diabetes in a subject by administering to the subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure. In one aspect, the subject suffers from diabetic nephropathy. In another aspect, the subject suffers from renal insufficiency. In another aspect, administration of the compound is an adjunct to diet and exercise. In another aspect, administration of the compound also reduces body weight and / or treats obesity. In another aspect, the subject has a 27 kg / m 2 Up to 40kg / m 2 In another aspect, the subject has a BMI of 30 kg / m 2 Up to 39.9kg / m 2 In another aspect, the subject has a BMI of at least 40 kg / m 2 In another aspect, the subject is overweight. In another aspect, the subject is obese.

[0344] In some embodiments, the present disclosure provides a method of improving glycemic control in a subject by administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure. In one aspect, the subject suffers from type 2 diabetes. In another aspect, the subject suffers from diabetic nephropathy. In another aspect, the subject suffers from renal insufficiency. In another aspect, administration of the compound is an adjunct to diet and exercise. In another aspect, administration of the compound also reduces body weight and / or treats obesity. In another aspect, the subject has a 27 kg / m 2 Up to 40kg / m 2 In another aspect, the subject has a BMI of 30 kg / m 2 Up to 39.9kg / m 2In another aspect, the subject has a BMI of at least 40 kg / m 2 In another aspect, the subject is overweight. In another aspect, the subject is obese.

[0345] In some embodiments, the present disclosure provides a method of improving glycemic control in a subject by administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure. In one aspect, administration of the compound is an adjunct to diet and exercise. In another aspect, administration of the compound also reduces body weight and / or treats obesity. In another aspect, the subject has a 27 kg / m 2 Up to 40kg / m 2 In another aspect, the subject has a BMI of 30 kg / m 2 Up to 39.9kg / m 2 In another aspect, the subject has a BMI of at least 40 kg / m 2 In another aspect, the subject is overweight. In another aspect, the subject is obese.

[0346] In some embodiments, the present disclosure provides a method of improving glycemic control in a subject suffering from type 2 diabetes and renal insufficiency by administering a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure to the subject. In one aspect, administration of the compound is an adjunct to diet and exercise. In another aspect, administration of the compound also reduces body weight and / or treats obesity. In another aspect, the subject has 27 kg / m 2 Up to 40kg / m 2 In another aspect, the subject has a BMI of 30 kg / m 2 Up to 39.9kg / m 2 In another aspect, the subject has a BMI of at least 40 kg / m 2 In another aspect, the subject is overweight. In another aspect, the subject is obese.

[0347] In some embodiments, the present disclosure provides a method for treating or preventing insulin resistance of a subject by administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure. In another aspect, the subject suffers from type 2 diabetes. In another aspect, the subject suffers from diabetic nephropathy. In another aspect, the subject suffers from renal insufficiency. For example, insulin resistance can be measured using the homeostatic model assessment (HOMA-IR) and / or the MATSUDA index of insulin resistance. For example, HOMA-IR is explained in Diabetologia 1985, 28, 412, which is incorporated herein by reference in its entirety. For example, the MATSUDA index is explained in Diabetes Care 1999, 22, 1462, which is incorporated herein by reference in its entirety.

[0348] In some embodiments, the present disclosure provides a method of treating or preventing glucose intolerance in a subject by administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure. In one aspect, the subject suffers from type 2 diabetes. In another aspect, the subject suffers from diabetic nephropathy. In another aspect, the subject suffers from renal insufficiency.

[0349] In some embodiments, the present disclosure provides a method of treating a subject's cardiovascular disorder by administering a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure to a subject in need of treatment. In one aspect, the cardiovascular disorder is selected from the group consisting of heart failure, cardiomyopathy, atherosclerosis, venous thromboembolism, and atrial fibrillation. In one aspect, the cardiovascular disorder is heart failure. In another aspect, the cardiovascular disorder is heart failure with preserved ejection fraction (HFpEF). In another aspect, the cardiovascular disorder is cardiomyopathy. In another aspect, the cardiomyopathy is selected from the group consisting of hypertrophic cardiomyopathy, dilated cardiomyopathy, restrictive cardiomyopathy, hypertrophic cardiomyopathy, ischemic cardiomyopathy, ischemic cardiomyopathy, dilated cardiomyopathy, and idiopathic cardiomyopathy. In another aspect, the cardiovascular disorder is atherosclerosis. In another aspect, the cardiovascular disorder is venous thromboembolism. In another aspect, the cardiovascular disorder is atrial fibrillation.

[0350] In some embodiments, the present disclosure provides a method for treating obesity or obesity-related disorders of a subject by administering a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure to a subject in need thereof. In one aspect, obesity-related disorders are obesity-related metabolic disorders. In another aspect, obesity-related disorders are selected from the group consisting of insulin resistance, prediabetes, type 2 diabetes, glucose intolerance, elevated fasting blood sugar, and glucagonoma. In another aspect, obesity-related disorders are dyslipidemia. In another aspect, obesity-related disorders are cardiovascular disorders selected from the group consisting of heart failure, cardiomyopathy, atherosclerosis, venous thromboembolism, and atrial fibrillation. In another aspect, obesity-related disorders are kidney disease.

[0351] In some embodiments, the present disclosure provides a method of reducing the weight of a subject by administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure. In one aspect, the subject suffers from type 2 diabetes. In another aspect, the subject suffers from diabetic nephropathy. In another aspect, the subject suffers from renal insufficiency. In another aspect, administration of the compound is an adjunct to diet and exercise. In another aspect, administration of the compound also reduces weight and / or treats obesity. In another aspect, the subject has a 27 kg / m 2 Up to 40kg / m 2 In another aspect, the subject has a BMI of 30 kg / m 2 Up to 39.9kg / m 2 In another aspect, the subject has a BMI of at least 40 kg / m 2 In another aspect, the subject is overweight. In another aspect, the subject is obese. In another aspect, the subject has a reduction in body weight, e.g., at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, or 40%.

[0352] In some embodiments, the present disclosure provides a method of reducing body fat in a subject by administering to a subject in need of treatment a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure. In another aspect, the subject suffers from type 2 diabetes. In another aspect, the subject suffers from diabetic nephropathy. In another aspect, the subject suffers from renal insufficiency. In another aspect, administration of the compound is an adjunct to diet and exercise. In another aspect, administration of the compound also reduces body weight and / or treats obesity. In another aspect, the subject has 27 kg / m 2 Up to 40kg / m 2 In another aspect, the subject has a BMI of 30 kg / m 2 Up to 39.9kg / m 2In another aspect, the subject has a BMI of at least 40 kg / m 2 In another aspect, the subject is overweight. In another aspect, the subject is obese. In another aspect, the fat is liver fat.

[0353] In some embodiments, the present disclosure provides a method for treating or preventing fibrosis in a subject by administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure. In one aspect, the fibrosis is interstitial lung disease. In another aspect, the fibrosis is interstitial lung disease with progressive fibrosis. In another aspect, the interstitial lung disease is pulmonary fibrosis. In another aspect, the interstitial lung disease is idiopathic pulmonary fibrosis (IPF).

[0354] In some embodiments, the present disclosure provides a method for promoting tissue remodeling in a subject by administering to the subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure. In one aspect, the subject suffers cardiac tissue damage due to myocardial infarction.

[0355] In some embodiments, the present disclosure provides a method of promoting wound healing and / or reducing adhesions in a subject by administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure. In one aspect, administration of the crystalline form promotes wound healing and / or reduces adhesions by increasing fibrinolysis.

[0356] In some embodiments, the present disclosure provides a method for treating or preventing a keloid disorder in a subject in need thereof by administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure. In one aspect, the keloid disorder is selected from the group consisting of scarring, keloid tumors, and keloid scars.

[0357] In some embodiments, the present disclosure provides a method for treating or preventing inflammation in a subject by administering to a subject in need thereof a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure. In one aspect, the inflammation is a chronic inflammation. In one aspect, the chronic inflammation is selected from the group consisting of rheumatoid arthritis, osteoarthritis, and Crohn's disease. In another aspect, the chronic inflammation is rheumatoid arthritis.

[0358] In some embodiments, the present disclosure provides a method of treating cancer in a subject by administering a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure to a subject in need of treatment. In one aspect, the cancer is selected from the group consisting of: breast cancer, pancreatic cancer, small intestine cancer, colon cancer, rectal cancer, lung cancer, head and neck cancer, ovarian cancer, hepatocellular carcinoma, esophageal cancer, hypopharyngeal cancer, nasopharyngeal cancer, laryngeal cancer, myeloma cells, bladder cancer, bile duct cell carcinoma, clear cell renal carcinoma, neuroendocrine tumor, carcinogenic osteomalacia, sarcoma, CUP (unknown primary cancer), thymic carcinoma, ligamentous tumor, glioma, astrocytoma, cervical cancer and prostate cancer. In another aspect, the cancer is hepatocellular carcinoma.

[0359] The subject of treatment is usually a human or non-human mammal, especially a human. Suitable subjects may also include domestic or wild animals; companion animals (including dogs, cats, etc.); livestock (including horses, cattle and other ruminants, pigs, poultry, rabbits, etc.); primates (including monkeys, such as rhesus monkeys, macaques (also known as crab monkeys or long-tailed monkeys), marmosets, silk monkeys, chimpanzees, macaques, etc.); and rodents (including rats, mice, gerbils, guinea pigs, etc.).

[0360] In some embodiments, the present disclosure provides a crystalline form of the present disclosure for use as a medicament.

[0361] In some embodiments, the present disclosure provides for use of a crystalline form of the present disclosure for treating or preventing a FAP-mediated disorder as discussed above.

[0362] In some embodiments, the present disclosure provides for the use of a crystalline form of the present disclosure for the manufacture of a medicament for the treatment or prevention of a FAP-mediated disorder as discussed above.

[0363] IV. Combination Therapies and Fixed-Dose Combinations

[0364] The crystalline forms of the present disclosure can be used in the above methods as a single pharmacological agent or in combination with other pharmacological agents or technologies. Such combination therapies can be achieved by administering the components of the treatment simultaneously, sequentially or separately. These combination therapies (and corresponding combination products) employ the crystalline forms of the present disclosure within the dosage range described in this specification and other pharmacological agents generally within their approved dosage ranges.

[0365] In some embodiments, the present disclosure provides a combination suitable for treating a disease selected from the previously discussed disease, wherein the combination comprises a crystalline form of the present disclosure and a sodium-glucose transporter 2 (SGLT2) inhibitor. In one aspect, the SGLT2 inhibitor is selected from the group consisting of canagliflozin, dapagliflozin, empagliflozin, erepagliptin, ipagliptin, lupagliflozin and remogliflozin. In another aspect, the SGLT2 inhibitor is dapagliflozin.

[0366] In some embodiments, the present disclosure provides a combination useful for treating a condition selected from the conditions previously discussed, wherein the combination comprises a crystalline form of the present disclosure and metformin.

[0367] In some embodiments, the present disclosure provides a combination suitable for treating a condition selected from the previously discussed conditions, wherein the combination comprises a crystalline form of the present disclosure and a glucagon-like peptide-1 receptor (GLP1) agonist. In one aspect, the GLP1 agonist is selected from the group consisting of exenatide, liraglutide, lixisenatide, albiglutide, dulaglutide and semaglutide.

[0368] In some embodiments, the present disclosure provides a combination suitable for treating a condition selected from the previously discussed conditions, wherein the combination comprises a crystalline form of the present disclosure and a dipeptidyl peptidase 4 (DPP4) inhibitor. In one aspect, the DPP4 inhibitor is selected from the group consisting of sitagliptin, vildagliptin, saxagliptin, linagliptin, gemagliptin, alagliptin, teneligliptin, alogliptin, trelagliptin, ogliptin, emagliptin, gogliptin and dulagliptin.

[0369] In some embodiments, the present disclosure provides a combination suitable for treating a disorder selected from the disorders discussed previously, wherein the combination comprises a crystalline form of the present disclosure and a peroxisome proliferator-activated receptor (PPAR) agonist. In one aspect, the PPAR agonist is a PPARα agonist. In another aspect, the PPAR agonist is a PPARγ agonist. In another aspect, the PPAR agonist is a PPARα / γ agonist. In another aspect, the PPAR agonist is selected from the group consisting of clofibrate, gemfibrozil, ciprofibrate, bezafibrate and fenofibrate. In another aspect, the PPAR agonist is a thiazolidinedione. In another aspect, the thiazolidinedione is selected from the group consisting of pioglitazone, rosiglitazone, lobeglitazone and rivoglitazone. In another aspect, the PPAR agonist stimulates the liver expression of FGF21.

[0370] In some embodiments, the present disclosure provides a pharmaceutical composition comprising a crystalline form of the present disclosure; one or more pharmacological agents selected from the group consisting of SGLT2 inhibitors, metformin, GLP1 agonists, DPP4 inhibitors and PPAR agonists; and a pharmaceutically acceptable diluent or carrier. Such a combination can be used to manufacture a medicament for treating a condition selected from the conditions discussed previously. In one aspect, the pharmaceutical composition comprises an SGLT2 inhibitor. In another aspect, the pharmaceutical composition comprises metformin. In another aspect, the pharmaceutical composition comprises a GLP1 agonist. In another aspect, the pharmaceutical composition comprises a DPP4 inhibitor. In another aspect, the pharmaceutical composition comprises a PPAR agonist.

[0371] In some embodiments, the present disclosure provides a combination suitable for treating cancer, wherein the combination comprises a crystalline form of the present disclosure and an immune checkpoint inhibitor. In one aspect, the immune checkpoint inhibitor is selected from the group consisting of anti-PD-1 antibodies, anti-PD-L1 antibodies, anti-CTLA4 antibodies, TLR7 agonists, CD40 agonists, Lag3 antagonists and OX40 agonists. In another aspect, the immune checkpoint inhibitor is an anti-PD-1 antibody (e.g., pembrolizumab (Keytruda), nivolumab (Opdivo), cemiplimab (Libtayo), etc.). In another aspect, the immune checkpoint inhibitor is an anti-PD-L1 antibody (e.g., atezolizumab (Tecentriq), avelumab (Bavencio), durvalumab (Imfinzi), etc.). In another aspect, the immune checkpoint inhibitor is an anti-CTLA4 antibody (e.g., ipilimumab (Yervoy), tremelimumab, etc.). In another aspect, cancer is selected from the group consisting of pancreatic cancer, colon cancer and rectal cancer.

[0372] V. Pharmaceutical Compositions

[0373] The crystalline form of the present disclosure may be administered as a pharmaceutical composition comprising one or more pharmaceutically acceptable excipients. Thus, in some embodiments, the present disclosure provides a pharmaceutical composition comprising the crystalline form of the present disclosure and at least one pharmaceutically acceptable excipient.

[0374] The excipient selected to be included in a particular composition will depend on factors such as the form of the mode of administration and the composition provided. Suitable pharmaceutically acceptable excipients are well known to those skilled in the art, and are described in, for example, Handbook of Pharmaceutical Excipients, the sixth edition, Pharmaceutical Press, edited by Rowe, Ray C, Sheskey, Paul J, Quinn, Marian. Pharmaceutically acceptable excipients can be used as, for example, adjuvants, diluents, carriers, stabilizers, flavoring agents, coloring agents, fillers, binders, disintegrants, lubricants, glidants, thickeners and coating agents. As will be appreciated by those skilled in the art, some pharmaceutically acceptable excipients can play more than one role, and can play a substitution role, depending on how many excipients there are in the composition and which other excipients there are in the composition.

[0375] The composition may be in a form suitable for oral use (e.g. as tablets, lozenges, hard or soft capsules, aqueous or oily suspensions, emulsions, dispersible powders or granules, syrups or elixirs), in a form suitable for topical use (e.g. as creams, ointments, gels, or aqueous or oily solutions or suspensions), in a form suitable for administration by inhalation (e.g. as finely divided powders or liquid aerosols), in a form suitable for administration by insufflation (e.g. as finely divided powders) or in a form suitable for parenteral administration (e.g. as sterile aqueous or oily solutions for intravenous, subcutaneous or intramuscular administration), or as suppositories for rectal administration. The composition may be obtained by conventional procedures using conventional pharmaceutical excipients well known in the art. Thus, compositions intended for oral use may contain, for example, one or more coloring agents, sweeteners, flavoring agents and / or preservatives.

[0376] The total daily dose will necessarily vary depending on the subject being treated, the route of administration, any co-administered therapy, and the severity of the disease being treated, and may include single or multiple doses. The specific dose may be adjusted, for example, according to: the condition being treated; the age, weight, general health, sex, and diet of the subject; the route of administration; the dosage interval; the excretion rate; and other drugs co-administered to the subject. A person of ordinary skill provided with the disclosure of this specification will be able to determine appropriate doses and regimens for administering the therapeutic agent to a subject according to methods well known in the therapeutic art, and adjust such doses and regimens as necessary during the course of treatment. The crystalline form of the present disclosure will generally be administered at 2.5 mg / m 2 Animal body area up to 5000mg / m 2 A unit dosage in the range of about 0.05 mg / kg to 100 mg / kg per animal body area or animal is administered to a warm-blooded animal, and this generally provides a therapeutically effective dose.

[0377] In some embodiments, the present disclosure provides a pharmaceutical composition for use in therapy, comprising a crystalline form of the present disclosure and at least one pharmaceutically acceptable excipient.

[0378] In some embodiments, the present disclosure provides a pharmaceutical composition for use in treating a FAP-mediated disorder, comprising a crystalline form of the present disclosure and at least one pharmaceutically acceptable excipient. In one aspect, the FAP-mediated disorder is selected from the group consisting of liver disease, type 2 diabetes, cardiovascular disorders, obesity, obesity-related disorders, fibrosis, keloid disorders, inflammation, and cancer.

[0379] In some embodiments, the present disclosure provides a pharmaceutical composition comprising Form A and one or more pharmaceutically acceptable excipients. In one aspect, the composition comprises at least 90% by weight of Form A relative to any other crystalline form. In another aspect, the composition comprises at least 95% by weight of Form A relative to any other crystalline form. In another aspect, the composition comprises at least 96% by weight of Form A relative to any other crystalline form. In another aspect, the composition comprises at least 97% by weight of Form A relative to any other crystalline form. In another aspect, the composition comprises at least 98% by weight of Form A relative to any other crystalline form. In another aspect, the composition comprises at least 99% by weight of Form A relative to any other crystalline form.

[0380] In some embodiments, the present disclosure provides a pharmaceutical composition comprising Form B and one or more pharmaceutically acceptable excipients. In one aspect, the composition comprises at least 90% by weight of Form B relative to any other crystalline form. In another aspect, the composition comprises at least 95% by weight of Form B relative to any other crystalline form. In another aspect, the composition comprises at least 96% by weight of Form B relative to any other crystalline form. In another aspect, the composition comprises at least 97% by weight of Form B relative to any other crystalline form. In another aspect, the composition comprises at least 98% by weight of Form B relative to any other crystalline form. In another aspect, the composition comprises at least 99% by weight of Form B relative to any other crystalline form.

[0381] In some embodiments, the present disclosure provides a pharmaceutical composition comprising Form A and Form B and one or more pharmaceutically acceptable excipients. In one aspect, the composition comprises at least 50% by weight of Form B relative to Form A. In another aspect, the composition comprises at least 75% by weight of Form B relative to Form A. In another aspect, the composition comprises at least 90% by weight of Form B relative to Form A. In another aspect, the composition comprises at least 95% by weight of Form B relative to Form A. In another aspect, the composition comprises at least 99% by weight of Form B relative to Form A.

[0382] In some embodiments, the present disclosure provides a pharmaceutical composition comprising Type 2 and one or more pharmaceutically acceptable excipients. In one aspect, the composition comprises at least 90% by weight of Type 2 relative to any other crystalline form. In another aspect, the composition comprises at least 95% by weight of Type 2 relative to any other crystalline form. In another aspect, the composition comprises at least 96% by weight of Type 2 relative to any other crystalline form. In another aspect, the composition comprises at least 97% by weight of Type 2 relative to any other crystalline form. In another aspect, the composition comprises at least 98% by weight of Type 2 relative to any other crystalline form. In another aspect, the composition comprises at least 99% by weight of Type 2 relative to any other crystalline form.

[0383] VI. Kit

[0384] The disclosure also provides kits comprising a unit dosage form including a crystalline form of the disclosure contained within packaging material and a label or package insert indicating that the unit dosage form can be used to treat one or more of the previously described disorders.

[0385] In some embodiments, the kit comprises a unit dosage form including a crystalline form of the disclosure contained within a packaging material and a label or package insert indicating that the pharmaceutical composition can be used to treat a FAP-mediated disorder. In one aspect, the FAP-mediated disorder is a liver disease. In another aspect, the liver disease is selected from the group consisting of fatty liver disease, end-stage liver disease, and cirrhosis. In another aspect, the liver disease is selected from the group consisting of non-alcoholic steatohepatitis (NASH) and non-alcoholic fatty liver disease (NAFLD).

[0386] In some embodiments, the kit comprises: (a) a first unit dosage form comprising a crystalline form of the present disclosure; (b) a second unit dosage form comprising a pharmacological agent selected from the group consisting of an SGLT2 inhibitor, metformin, a GLP1 agonist, a DPP4 inhibitor, and a PPAR agonist; (c) a container device for containing the first dosage form and the second dosage form; and (d) a label or package insert indicating that the first unit dosage form and the second unit dosage form can be used to treat a FAP-mediated disorder.

[0387] VII. Examples

[0388] The following descriptions of experiments, procedures, examples, and intermediates are intended to illustrate embodiments of the present disclosure. They are by no means limiting. Other embodiments of the present disclosure may be prepared using the methods described in these examples, either alone or in combination with techniques generally known in the art.

[0389] Example 1: (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxylate Preparation of amines

[0390]

[0391] (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide can be prepared as follows:

[0392] A. (R)-3-Glycylthiazolidine-4-carbonitrile hydrochloride (Intermediate 1)

[0393]

[0394] (i) (R)-4-Carbamoylthiazolidine-3-carboxylic acid tert-butyl ester (Intermediate 1-A)

[0395]

[0396] Di-tert-butyl dicarbonate (BocO, 18.6mL, 80.2mmol) is added to (R)-3-(tert-butoxycarbonyl) thiazolidine-4-carboxylic acid (17.0g, 72.9mmol) and pyridine (7.07mL, 87.5mmol) in a stirred solution in ethyl acetate (170mL), and the reaction mixture is stirred at room temperature for 3 hours. NH is then added dropwise solution (25% aqueous solution, 6mL), and the mixture is stirred at room temperature overnight. The reaction mixture is diluted with ethyl acetate, each phase is separated, and the organic phase is washed with saturated NaCl, dried, filtered through a silica gel pad, washed with ethyl acetate, and evaporated to obtain the crude (R)-4-carbamoyl thiazolidine-3-carboxylic acid tert-butyl ester (intermediate 1-A, 16.9g, 100%) as a colorless oil, which is directly used in the next step.

[0397] (ii) (R)-4-cyanothiazolidine-3-carboxylic acid tert-butyl ester (Intermediate 1-B)

[0398]

[0399] At room temperature, trifluoroacetic anhydride (TFAA, 12.4mL, 87.5mmol) solution in ethyl acetate (20mL) is added to crude (R)-4-carbamoyl-thiazolidine-3-carboxylic acid tert-butyl ester (intermediate 1-A, 16.9g, 72.9mmol) and pyridine (14.7mL, 182mmol) in ethyl acetate (150mL).Mixture is stirred at room temperature 4 hours, and then diluted with ethyl acetate, with HCl aqueous solution (1M) and saturated NaHCO washing.Organic phase is dried, filtered through silica gel pad, washed with ethyl acetate, and evaporation obtains light yellow oil, which solidifies when standing.Crude solid matter is suspended in heptane: in ethyl acetate (4:1,50mL), and at room temperature stirred overnight. The solid was filtered off, washed with heptane:ethyl acetate (4:1), and dried to give (R)-tert-butyl 4-cyanothiazolidine-3-carboxylate (Intermediate 1-B, 12.0 g, 83%) as a colorless solid; 1 H NMR (400MHz, CDCl3) δ5.20–4.79(m,1H),4.60–4.53(m,1H),4.53–4.36(m,1H),3.40-3.18(m,2H),1.51(s,9H).

[0400] (iii) (R)-Thiazolidine-4-carbonitrile hydrochloride (Intermediate 1-C)

[0401]

[0402] At room temperature, a solution of aqueous HCl (12M, 11 mL) in methanol (140 mL) was slowly added to a solution of (R)-4-cyanothiazolidine-3-carboxylic acid tert-butyl ester (Intermediate 1-B, 6.0 g, 28 mmol) in methanol (140 mL). The clear colorless solution was stirred at room temperature for 2 hours. The solvent was evaporated to give (R)-thiazolidine-4-carbonitrile hydrochloride (Intermediate 1-C, 4.22 g, 100%) as a colorless solid; 1 H NMR (400MHz, CD3OD) δ4.90(dd,1H), 4.35–4.24(m,2H), 3.37–3.24(m,2H).

[0403] (iv) tert-Butyl (R)-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)carbamate (Intermediate 1-D)

[0404]

[0405] N-ethyl-N-isopropyl-propan-2-amine (DIPEA, 19.6 mL, 112 mmol) was added to a suspension of (R)-thiazolidine-4-carbonitrile hydrochloride (Intermediate 1-C, 4.22 g, 28 mmol), (tert-butoxycarbonyl)glycine (6.13 g, 35.0 mmol) and propanephosphonic anhydride (T3P, 41.6 mL, 70.0 mmol, 50% ethyl acetate solution) in ethyl acetate (120 mL). The mixture was heated at 60° C. for 4 hours. The mixture was diluted with ethyl acetate and washed sequentially with water, aqueous HCl solution (1 M) and saturated NaHCO3. The organic phase was dried, filtered and evaporated. The residue was filtered through a pad of silica gel, washed with heptane:ethyl acetate (1:1) and evaporated to give an oil, which was triturated with heptane:DCM to give tert-butyl (R)-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)carbamate (Intermediate 1-D, 7.60 g, 100%) as an almost colorless solid; 1 H NMR (400MHz, CDCl3) δ5.36–5.25(m,2H),4.59–4.52(m,2H),4.14–3.90(m,2H),3.29(d,2H),1.45(s,9H).

[0406] (v) (R)-3-Glycylthiazolidine-4-carbonitrile hydrochloride (Intermediate 1)

[0407]

[0408] A solution of aqueous HCl (12M, 5.6 mL) in methanol (140 mL) was slowly added to a solution of (R)-tert-butyl (2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)carbamate (Intermediate 1-D, 7.60 g, 28.0 mmol) in methanol (140 mL), and the solution was stirred at room temperature overnight. The solvent was evaporated to give (R)-3-glycylthiazolidine-4-carbonitrile hydrochloride (Intermediate 1, 5.80 g, 100%) as a colorless solid; 1 H NMR (400MHz, CD3OD) δ5.34(t,1H),4.72(d,1H),4.62(d,1H),4.11–3.94(m,2H),3.41–3.36(m,2H).

[0409] B. 6-Morpholinoquinoline-4-carboxylic acid (Intermediate 2)

[0410]

[0411] (i) Ethyl 6-morpholinoquinoline-4-carboxylate (Intermediate 2-A)

[0412]

[0413] Morpholine (0.22 mL, 2.5 mmol) was added to a mixture of ethyl 6-bromoquinoline-4-carboxylate (0.355 g, 1.27 mmol), bis(dibenzylideneacetone)palladium (Pd(dba)2, 36 mg, 0.06 mmol), dicyclohexyl(2',6'-diisopropoxy-[1,1'-biphenyl]-2-yl)phosphane (RuPhos, 59 mg, 0.13 mmol) and K3PO4 (0.538 g, 2.53 mmol) in 2-methylpropan-2-ol (2.3 mL). The flask was sealed, purged with N2(g), and heated at 90°C overnight. The reaction mixture was diluted with ethyl acetate and washed sequentially with water and brine. The organic layer was dried by a phase separator and concentrated under reduced pressure to give ethyl 6-morpholinoquinoline-4-carboxylate (Intermediate 2-A, 110 mg, 30%); MS m / z (ESI), [M+H]+ 287.2.

[0414] (ii) 6-Morpholinoquinoline-4-carboxylic acid (Intermediate 2)

[0415]

[0416] To a solution of ethyl 6-morpholinoquinoline-4-carboxylate (Intermediate 2-A, 110 mg, 0.38 mmol) in methanol (4 mL) was added NaOH (31 mg, 0.77 mmol) and heated at 60 ° C for 2 hours. The reaction mixture was cooled to room temperature and aqueous HCl (0.023 mL) was added. The reaction mixture was concentrated under reduced pressure to give 6-morpholinoquinoline-4-carboxylic acid (Intermediate 2, 95 mg, 96%); MS m / z (ESI), [M+H]+259.1.

[0417] C. (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide

[0418]

[0419] N-ethyl-N-isopropyl-propan-2-amine (DIPEA, 0.15mL, 0.87mmol) is added to a suspension of 6-morpholinoquinoline-4-carboxylic acid (intermediate 2, 75mg, 0.29mmol), (R)-3-glycylthiazolidine-4-carbonitrile hydrochloride (intermediate 1, 121mg, 0.58mmol), 1-hydroxybenzotriazole hydrate (HOBt, 53mg, 0.35mmol) and 3-(ethyliminomethyleneamino)-N,N-dimethyl-propan-1-amine hydrochloride (EDC, 84mg, 0.44mmol) in ethyl acetate (1mL) and acetonitrile (1mL). A clear yellow solution is obtained, which is stirred at room temperature overnight. The mixture is diluted with ethyl acetate and washed with saturated NaHCO and brine. The organic phase is dried, filtered and evaporated. The residue was purified by preparative HPLC on a Kromasil C8 column (10 μm, 250×20 mm ID) using a gradient of acetonitrile in H 2 O / acetonitrile / formic acid (95 / 5 / 0.2) (5%-65%) as the mobile phase to give (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide (26 mg, 21%); HRMS (ESI) m / z: for C 20 H 22 N5O3S[M+H]+ Calculated value: 412.1438 Found: 412.1437; 1 H NMR(400MHz,CD3CN)δ8.71(d,1H),8.05(s,1H),7.96(d,1H),7.69(d,1H),7.60(dd,1H),7.43 (d,1H),5.24(dd,1H),4.79–4.60(m,2H),4.30(d,2H),3.88–3.79(m,4H),3.35–3.28(m,6H).

[0420] Example 2: Analytical Methods

[0421] Unless otherwise stated, the following analytical methods were used to characterize the crystalline forms described in the following examples:

[0422] A. Reflection Powder X-ray Diffraction (Amorphous PXRD)

[0423] X-ray diffraction analysis was performed according to standard procedures, which can be found in, for example, Kitaigorodsky, AI (1973), Molecular Crystals and Molecules, Academic Press, New York; Bunn, CW (1948), Chemical Crystallography, Clarendon Press, London; or Klug, HP and Alexander, LE (1974), X-ray Diffraction Procedures, John Wiley & Sons, New York.

[0424] Powder X-ray diffraction (PXRD) patterns were determined by mounting the sample on a zero-background holder, single-crystal silicon, and developing the sample into thin layers. PXRD (X-ray wavelength -nickel filtered Cu radiation, voltage 45 kV, filament emission 40 mA). Variable divergence and anti-scatter slits and incident and diffraction Soller slits of 0.04° were used. The sample was rotated during the measurement. The sample was scanned from 2.4-50° 2θ using a 0.013° step size and 115.770 s counting time with a PIXcel 1D detector (effective length 3.35° 2θ). The PXRD pattern was obtained in Bragg-Brentano geometry.

[0425] Those skilled in the art will recognize that, depending on the measurement conditions, such as the equipment or machine used, a PXRD pattern with one or more measurement errors may be obtained (Jenkins, R & Snyder, RL 'Introduction to X-Ray Powder Diffractometry' John Wiley & Sons 1996; Bunn, CW (1948), Chemical Crystallography, Clarendon Press, London; Klug, HP & Alexander, LE (1974), X-Ray Diffraction Procedures). Those skilled in the art of X-ray powder diffraction will further recognize that the relative intensity of the peaks may be affected by, for example, grains with a size greater than 30 microns and non-single aspect ratios that may affect sample analysis. In addition, it should be understood that the intensity may fluctuate depending on the experimental conditions and sample preparation (e.g., preferred orientation). The following definitions are used for relative intensity (%): 25%-100%, vs (very strong); 10%-25%, s (strong); 3%-10%, m (medium); 1%-3%, w (weak).

[0426] Those skilled in the art will also recognize that the position of the reflections may be affected by the precise height at which the sample is located in the diffractometer and the zero calibration of the diffractometer. The surface flatness of the sample may also have a small effect. Therefore, the diffraction pattern data presented are not to be considered as absolute values. Typically, the measurement error of the diffraction angles in an X-ray powder diffraction pattern can be about plus or minus 0.2° 2θ, and this degree of measurement error should be taken into account when considering PXRD data.

[0427] Reflection mode PXRD patterns can be compared to transmission mode PXRD patterns, but one skilled in the art will recognize that the diffraction patterns may vary, particularly with respect to peak intensity.

[0428] B. Reflection Powder X-ray Diffraction (Type 2 PXRD)

[0429] Powder X-ray diffraction (PXRD) patterns were determined by mounting the sample on a zero-background holder, single-crystal silicon, and developing the sample into thin layers. Powder X-ray diffraction (X-ray wavelength -nickel-filtered Cu radiation, voltage 45 kV, filament emission 40 mA). Variable divergence and anti-scatter slits as well as entrance and diffraction Soller slits of 0.04° were used. The sample was rotated during the measurement. The sample was scanned from 2.4–50° 2θ using a 0.013° step size and 234.345 s counting time with a PIXcel3D-Medipix3 detector (effective length 3.35° 2θ). The following definitions are used for relative intensities (%): 25%-100%, vs (very strong); 10%-25%, s (strong); 3%-10%, m (medium); 1%-3%, w (weak).

[0430] C. Transmission Powder X-ray Diffraction (PXRD of Form A and Form B)

[0431] Powder X-ray diffraction data were measured using corundum as an internal reference. Powder X-ray diffraction (PXRD) patterns were obtained by mounting the sample on two The polyimide films are formed between the films to determine the thin layer. PXRD (X-ray wavelength) was recorded using a transmission PANalytical Empyrean -nickel filtered Cu radiation, voltage 45 kV, filament emission 40 mA). Fixed divergence and anti-scatter slits were used, and the sample was rotated during the measurement. The sample was scanned from 2.5-50° 2θ using a 0.013° step size and 938.145 s counting time with a PIXcel3D-Medipix3 detector (effective length 3.35°θ). The following definitions are used for relative intensity (%): 25%-100%, vs (very strong); 10%-25%, s (strong); 3%-10%, m (medium); 1%-3%, w (weak).

[0432] D. Solid 13 C NMR spectroscopy

[0433] Approximately 100 mg of material was loaded into a 4 mm zirconium dioxide rotor sealed with a Kel-F cap. 13 C cross-polarization magic angle spinning spectrum, the rotor was rotated at 15 kHz (to remove chemical shift anisotropy) and recorded using cross-polarization from hydrogen 13 C spectra (to improve sensitivity and reduce experimental time). The contact time for magnetization transfer was 2 milliseconds and the inter-pulse delay (to allow nuclear relaxation) was 15 seconds. Using signal averaging, enough scans were recorded to make all major peaks distinguishable from the noise (typically 1 k scans). Glycine was used as a secondary chemical shift reference (major peak at 176.03 ppm).

[0434] E. Ramp Differential Scanning Calorimetry (DSC)

[0435] The melting point onset (T) was determined by differential scanning calorimetry using a TA Instruments DSC (Model Q2000). m ). The sample (about 1 mg-3 mg) was weighed into an aluminum sample pan. The sample was filled to the bottom of the sample pan and a cover with a pinhole was used. The instrument was purged with nitrogen at 50 mL / min and data were collected between 25°C and 210°C-250°C using a heating rate of 10°C / min.

[0436] F. MDSC Differential Scanning Calorimetry (DSC)

[0437] The glass transition temperature (T) was determined by MSC differential scanning calorimetry using a TA Instruments DSC (Model Q2000). g ). The sample (about 5 mg) was weighed into an aluminum sample pan. The sample was filled to the bottom of the sample pan and a cover with a pinhole was used. The instrument was purged with nitrogen at 50 mL / min and data were collected between 25°C and 230°C using a heat-only modulation method with a base heating rate of 5°C / min and an adjustment amplitude of 0.53°C every 40 seconds.

[0438] G. Thermogravimetric analysis (TGA)

[0439] Thermogravimetric analysis was performed using a TA Instruments TGA (model Q500). The sample (approximately 10 mg) was transferred to a tared sample holder. The instrument was purged with nitrogen, oven 60 mL / min, and balance 40 mL / min, and data were collected between room temperature and 300° C. using a heating rate of 10° C. / min. During heating, buoyancy effects will result in the observed weight increase. This effect can be reduced by using more than 15 mg of material or performing a baseline subtraction on the sample curve.

[0440] H. Gravimetric Vapor Sorption (GVS)

[0441] Gravimetric vapor sorption analysis was performed using a TA Instruments TGA (Model Q5000SA). The sample (approximately 5 mg-10 mg) was transferred to a tared sample holder. The instrument was purged with nitrogen at 25°C, 200 mL / min in the chamber, and 10 mL / min on the balance, and data were collected at a unique peak relative humidity (%RH). Start at 20%RH and step up to 80%RH, step down to 0%RH, and finally a second cycle up to 90%RH and back to 0%RH. When the drift standard (dm / dt) was below 0.002 for 10 minutes, the equilibrium standard for moving to the next %RH was reached.

[0442] Hygroscopicity can be assessed, for example, according to the European Pharmacopoeia (EP) classification: non-hygroscopic: <0.2%; slightly hygroscopic: ≥0.2% and <2%; hygroscopic: ≥2% and <15%; very hygroscopic: ≥15%; deliquescent: absorbs enough water to form a liquid; all values ​​are measured as weight gain at 80% RH and 25°C.

[0443] Example 3: Form A / Form B Mixture

[0444] Initial crystallization efforts to separate pure Form A and pure Form B typically produce a mixture of Form A and Form B. Crystallization parameters such as temperature, time, and solvent composition affect the type and quality of the final crystalline product obtained.

[0445] A.50 / 50 mixture

[0446] In this study, when (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide was separated from a mixture of isopropanol and ethyl acetate, a mixture of Form A and Form B (approximately 50 / 50) was obtained. Specifically, 6-morpholinoquinoline-4-carboxylic acid (Intermediate 2 of Example 1, 177 g, 0.757 mol, 1.0 equiv) was added to a 5 L jacketed vessel. (R)-3-glycylthiazolidine-4-carboxamide (159 g, 1.2 equiv) and ethyl acetate (3500 mL) were charged to the reactor. A 50% wt / wt solution of propylphosphonic anhydride in ethyl acetate (1310 g, 3 equiv) was then charged to the reactor, followed by DIPEA (443 g, 5.002 equiv). The temperature was adjusted to 65 °C and the reaction mixture was stirred for 48 hours then cooled to 20 °C.

[0447] The reaction mixture was combined with the second reaction mixture (465 g in total) prepared in a similar manner, and loaded into a 50 L container for post-processing. Ethyl acetate (20 L) was loaded into the container, and the pH was adjusted to about 9 using 10 L saturated NaHCO aqueous solution. The resulting two-phase mixture was stirred at 20 ° C for 15 minutes, followed by layer separation. The aqueous phase was extracted with 10 L ethyl acetate, and stirred at 20 ° C for 15 minutes, followed by phase separation. The organic phase was washed with 10 L saturated NaHCO aqueous solution and stirred at 20 ° C for 15 minutes, followed by layer separation. All organic phases were combined and concentrated to about 5 volumes using a rotary evaporator at 45 ° C. The organic phase was passed through a 5 kg silica pad using isopropyl alcohol (15 L) as an eluent. The organic phase was then concentrated to about 5 volumes using a rotary evaporator at 45 ° C, the concentrated organic phase was exchanged with 10 L ethyl acetate and concentrated to about 8 volumes. The suspension was filtered, and the wet cake was washed with 2 L ethyl acetate. The wet cake was dried at 50°C for 18 hours to give a light yellow solid which was an approximately 50 / 50 mixture of Form A and Form B of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0448] B. Additional Studies

[0449] Small-scale crystallization studies were performed. Amorphous (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide was suspended or dissolved in each of the solvents listed in Table 1 below (20 relative volumes) and stirred at 20°C for 3 days. Each solid obtained was then analyzed by PXRD and DSC. The results are reported in Table 1 below.

[0450] Table 1

[0451]

[0452] The use of ethyl acetate or methyl isobutyl ketone as crystallization solvents appears to favor the formation of Form A, while the use of acetone, methyl ethyl ketone or acetonitrile appears to favor the formation of Form B. However, the other crystallization parameters selected also appear to affect the resulting crystalline form. In Example 3A above, a solvent system comprising isopropanol and ethyl acetate produced a 50 / 50 mixture of Form A and Form B. However, the low temperatures and limited time used during the concentration process may not be sufficient to dissolve Form A and convert it to Form B. In Example 5A(v) below, pure Form B was obtained starting from a 50 / 50 mixture of Form A and Form B using a solvent system comprising acetone and ethyl acetate. The suspension was stirred at elevated temperature for several hours, which may dissolve most of the lower melting Form A, and then enrich the suspension with the higher melting Form B over time, leaving Form B present to act as seed crystals when the suspension is cooled to 20°C.

[0453] Example 4: Crystalline Form A

[0454] A. Preparation of Crystalline Form A

[0455] Amorphous (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide (19 g) was suspended in ethyl acetate (about 450 mL) and the resulting fine suspension was heated to 75° C. A fine yellow suspension was obtained, which was slowly cooled to 20° C. over 5 hours and then stirred at 20° C. for another 12 hours. The fine suspension was filtered and the yellow solid was washed with ethyl acetate (2×50 mL). The collected solid was first relatively dry by suction at 20° C. under vacuum for 10 minutes and then at 40° C. under vacuum for 2 hours and finally at 35° C. under vacuum over the weekend. PXRD and DSC analysis showed mainly pure Form A.

[0456] B. Physical Characterization of Crystalline Form A

[0457] Form A was characterized using various techniques including PXRD ( Figure 1 ), solid state 13 C NMR spectroscopy ( Figures 2 to 5 )、Differential Scanning Calorimetry (DSC) ( Figure 6 )、Thermogravimetric analysis (TGA) ( Figure 7 ) and gravimetric vapor sorption (GVS) ( Figure 8 ).

[0458] Figure 1 The PXRD pattern confirmed that Form A is crystalline. Figure 1 The PXRD pattern of Form A measured using transmission geometry is shown in Table 2 below. Figure 1 Selected peaks identified in the PXRD pattern.

[0459] Table 2

[0460] Peak position (°2θ) Relative strength (%) definition 8.9 15 s 12.0 72 vs 13.1 12 s 14.4 10 m 17.5 22 s 17.9 17 s 18.4 36 vs 19.8 24 s 20.6 17 s 21.1 23 s 21.7 100 vs 23.6 10 m 25.6 6 m 27.3 6 m 31.2 7 m 39.9 6 m 42.0 6 m

[0461] Form A shows unique peaks at 12.0 ± 0.2 ° 2θ, 18.4 ± 0.2 ° 2θ, 19.8 ± 0.2 ° 2θ, and 21.7 ° 2θ ± 0.2 ° 2θ (relative to the other forms except Form B). Form A also shows characteristic peaks at 13.1 ± 0.2 ° 2θ, 14.4 ± 0.2 ° 2θ, 17.5 ± 0.2 ° 2θ, and 21.1 ° 2θ ± 0.2 ° 2θ. Form A lacks unique peaks at 18.7 ± 0.2 ° 2θ and 22.4 ° 2θ ± 0.2 ° 2θ relative to Form B (i.e., medium or stronger relative intensity peaks).

[0462] Figure 2 A representative solid state image showing a sample of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide 13 C NMR spectrum, this sample was approximately 95 wt % Form A and 5 wt % Form B. Figure 3 Representative solid state image showing a sample of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide 13 C NMR spectrum, this sample is approximately 30 wt % Form A and 70 wt % Form B. Figure 4 Representative solid state image showing a sample of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide 13 C NMR spectrum showed that the sample was Form B with a small amount of Form A. Figure 5 is based on Figure 2 , Figure 3 and Figure 4Comparison of solid state C NMR spectra of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Form A and Form B based on combined spectrum analysis of . Form A shows characteristic peaks at 168.0±0.2 ppm, 166.1±0.2 ppm, 147.5±0.2 ppm, 146.4±0.2 ppm, 143.1±0.2 ppm, 139.5±0.2 ppm, 130.7±0.2 ppm, 125±0.2 ppm, 117.9±0.2 ppm, 108.6±0.2 ppm, 67.1±0.2 ppm, 48.0±0.2 ppm, 42.9±0.2 ppm, and 35.2±0.2 ppm. Form A shows unique peaks relative to Form B at 166.1±0.2 ppm, 130.7±0.2 ppm, 117.9±0.2 ppm, 108.6±0.2 ppm, and 35.2±0.2 ppm.

[0463] Figure 6 Shown is a representative ramp DSC thermogram of Form A. Exothermic events are plotted in an upward direction. Figure 6 The melting endotherm shown has an onset temperature of about 171°C and an enthalpy of the melting endotherm of about 68 J / g. The DSC values ​​obtained can vary by as much as ±5°C, depending on the instrument used, how the sample was prepared, and batch-to-batch variations.

[0464] Figure 7 Shown is a representative TGA thermogram of Form A. Form A exhibits less than about 0.1% weight loss upon heating from about 25°C to 110°C, confirming that Form A is an anhydrate.

[0465] Figure 8 A representative GVS graph of Form A is shown. Form A exhibits a reversible moisture uptake of about 0.05 wt % between 0% relative humidity and 80% relative humidity at 25°C ± 0.1°C. The desorption curve shows that Form A loses moisture at a rate similar to that gained during adsorption, with limited hysteresis. No form change was observed by PXRD after the GVS experiment. Form A is non-hygroscopic (i.e., <0.2% weight gain) according to the European Pharmacopoeia (EP) classification.

[0466] Example 5: Crystalline Form B

[0467] A. Preparation of Crystalline Form B

[0468] (i) Unseeded crystals (acetone)

[0469] Amorphous (R)-N-(2-(4-cyanothiazolidine-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide (30 mg) was dissolved in 0.4 mL acetone. After 30 minutes, when the solution gradually thickened, more acetone (0.2 mL) was added to obtain a fine suspension. The suspension was stirred for three days. The fine suspension was filtered, and the solid was washed with acetone (0.6 mL). The solid was identified as Form B by DSC analysis.

[0470] (ii) Unseeded crystals (methyl ethyl ketone)

[0471] Amorphous (R)-N-(2-(4-cyanothiazolidine-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide (30 mg) was dissolved in 0.4 mL of methyl ethyl ketone. After 15 minutes, when the solution gradually thickened, more methyl ethyl ketone (0.2 mL) was added to obtain a fine suspension. The suspension was stirred for three days. The fine suspension was filtered, and the solid was washed with methyl ethyl ketone (0.6 mL). The solid was identified as Form B by DSC analysis.

[0472] (iii) Unseeded crystals (acetonitrile)

[0473] Amorphous (R)-N-(2-(4-cyanothiazolidine-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide (30 mg) was dissolved in 0.4 mL acetonitrile. After 45 minutes, when the solution gradually thickened, and after 2 hours, more acetonitrile (0.2 mL) was added to obtain a fine suspension. The suspension was stirred for three days. The fine suspension was filtered, and the solid was washed with acetonitrile (0.6 mL). The solid was identified as Form B by DSC analysis.

[0474] (iv) Seeded crystallization (acetonitrile)

[0475] (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide ("quinoline", 6.0 g) was dissolved in 90 mL of acetonitrile at 70°C, and the resulting solution was filtered through a glass microfiber filter. The filter was washed with 3.0 mL of acetonitrile, and the combined filtrates were cooled to 40°C at a rate of no more than 1.5°C / min. The solution was seeded with crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Form B (150 mg / 0.025 eq., pre-ground in a pestle and mortar), and then stirred at 40°C for at least 14 hours. The resulting slurry was cooled to 15°C at a rate of 5°C / hour, and the product was then isolated by vacuum filtration. The filter cake was washed successively with 6 mL of acetonitrile and 12 mL of tert-butyl methyl ether, and then dried under vacuum at 40°C to give the crystalline Form B final product (4.55 g).

[0476] The above procedure also worked well when the solution was seeded instead with crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Form B (90 mg / 0.015 equiv, pre-ground in pestle and mortar).

[0477] As described above, inoculation with 0.015 to 0.025 equivalents of ground Form B seeds worked well for acetonitrile solutions. Further modifications to the inoculation procedure were evaluated as described in Table 3 below:

[0478] Table 3

[0479]

[0480]

[0481] 1 The seeds were ground in portions and therefore may not be consistent across these experiments.

[0482] (v) Form A / Form B interconversion

[0483] Crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide (50 / 50 mixture of Form A and Form B, 20 g, 48.61 mmol) was charged to a reactor equipped with an overhead stirrer, followed by the addition of acetone (225 mL) and then ethyl acetate (75 mL). The resulting yellow suspension was slowly heated to reflux (T 夹套 =70°C) and stirred at this temperature (300 rpm) for 5 hours, then cooled to 20°C within 5 hours. The suspension was then stirred at 20°C for a further 15 hours. A very fine, easily stirred suspension was obtained with no solids attached to the glass walls of the reactor. The mixture was filtered and the solids were washed with 25% ethyl acetate in acetone (80 mL). The filter cake was then suction dried for 5 minutes. Further drying under vacuum at 40°C overnight gave 15 g of a yellow product. PXRD and DSC analysis (melting point 194°C) were consistent with Form B.

[0484] (vi) Seeded crystallization (methanol)

[0485] (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide (4.5 g) was dissolved in 100 mL of methanol at 60° C. The resulting solution was cooled to 45° C. at a rate of 5.0° C. / min. The solution was then seeded with crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Form B (0.0008 equivalents, pre-ground in a pestle and mortar) and stirred at 45° C. for 2 days. The solution was then cooled to 20° C. and maintained at 20° C. The most significant crystallization was observed approximately 2 days after cooling to 20° C. PXRD and DSC analysis were consistent with Form B, but additional studies (see Example 7) indicated that (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide may be chemically unstable in methanol.

[0486] (vii) Seeded crystallization (acetonitrile / 1-butanol)

[0487] The risk of racemization increases with the retention time of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide in acetonitrile solution at 70°C. To further reduce this risk, (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide (12.6 g) was dissolved in a mixture of acetonitrile (31.5 mL) and 1-butanol (58.3 mL) at 70°C, and the resulting solution was filtered through a glass microfiber filter. The filter was washed with a mixture of acetonitrile (1.1 mL) and 1-butanol (2.0 mL), and the combined filtrate was cooled to 44°C at a rate of 1.73°C / min. The solution was seeded with crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Form B (4.3 mg, 0.00035 equiv., pre-ground in a pestle and mortar) and then stirred at 44°C for at least 12 hours. The resulting slurry was cooled to 10°C at a rate of 4.9°C / hour and the product was then isolated by vacuum filtration. The filter cake was washed sequentially with 1-butanol (12.4 mL) and tert-butyl methyl ether (24.8 mL) and then dried under vacuum at 40°C. PXRD and DSC analysis were consistent with Form B.

[0488] B. Physical Characterization of Crystalline Form B

[0489] Form B was characterized using various techniques including PXRD ( Figures 9 to 11 ), solid state 13 C NMR spectroscopy ( Figures 2 to 5 )、Differential Scanning Calorimetry (DSC) ( Fig.12 )、Thermogravimetric analysis (TGA) ( Fig.13 ) and gravimetric vapor sorption (GVS) ( Fig.14 ).

[0490] Fig. 9 The PXRD pattern confirmed that Form B is crystalline. Fig. 9 The PXRD pattern of Form B measured using transmission mode is shown in Table 4 below. Fig. 9 Selected peaks identified in the PXRD pattern.

[0491] Table 4

[0492] Peak position (°2θ) Relative strength (%) definition 8.9 6 m 12.0 39 vs 13.1 7 m 14.4 6 m 17.5 14 s 18.0 7 m 18.4 22 s 18.7 19 s 19.8 34 vs 20.3 10 m 20.6 13 s 21.1 9.6 m 21.7 100 vs 22.4 30 vs 22.9 8 m 25.2 5 m 25.6 6 m 26.2 5 m 28.7 5 m 30.4 5 m 30.9 5 m 31.1 5 m

[0493] Form B shows unique peaks at 12.0 ± 0.2° 2θ, 18.4 ± 0.2° 2θ, 18.7 ± 0.2° 2θ, 19.8 ± 0.2° 2θ, 21.7 ± 0.2° 2θ, and 22.4° 2θ ± 0.2° 2θ (relative to the other forms except Form A). Form B also shows characteristic peaks at 13.1 ± 0.2° 2θ, 14.4 ± 0.2° 2θ, 17.5 ± 0.2° 2θ, 20.3 ± 0.2° 2θ, and 21.1° 2θ ± 0.2° 2θ.

[0494] Form B shows unique peaks relative to Form A at 18.7±0.2° 2θ and 22.4° 2θ±0.2° 2θ (ie, peaks of moderate or stronger relative intensity). Fig.10 is based on Figure 1 and Fig. 9 The diffraction patterns shown are comparisons of the transmission powder X-ray diffraction patterns of Form A and Form B. Fig.11 is a comparison of the transmission powder X-ray diffraction patterns of Form A and Form B covering the region of the unique Form B peak. The Form A diffraction pattern is Fig.10 and Fig.11 The top of the Form B diffraction pattern is Fig.10 and Fig.11 bottom.

[0495] As discussed previously for Form A, Figure 2 Representative solid state image showing a sample of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide 13 C NMR spectrum, this sample was approximately 95 wt % Form A and 5 wt % Form B. Figure 3 Representative solid state image showing a sample of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide 13C NMR spectrum, this sample is approximately 30 wt % Form A and 70 wt % Form B. Figure 4 Representative solid state image showing a sample of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide 13 C NMR spectrum showed that the sample was Form B with a small amount of Form A. Figure 5 is based on Figure 2 , Figure 3 and Figure 4 Combinatorial spectrum of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Form A and Form B in the solid state 13 Analysis of C NMR spectrum: Form B showed characteristic peaks at 167.7±0.2 ppm, 166.9±0.2 ppm, 147.3±0.2 ppm, 143.1±0.2 ppm, 139.9±0.2 ppm, 130.2±0.2 ppm, 125±0.2 ppm, 119.5±0.2 ppm, 118.6±0.2 ppm, 117.4±0.2 ppm, 110.0±0.2 ppm, 49.9±0.2 ppm, 46.6±0.2 ppm, and 34.5±0.2 ppm. Form B shows unique peaks relative to Form A at 166.9±0.2 ppm, 130.2±0.2 ppm, 118.6±0.2 ppm, 117.4±0.2 ppm, 110.0±0.2 ppm, 49.9±0.2 ppm, 46.6±0.2 ppm, and 34.5±0.2 ppm.

[0496] A representative ramp DSC thermogram of Form B is shown in Fig.12 Exothermic events are plotted in an upward direction. Fig.12 The melting endotherm shown has an onset temperature of about 191°C and a thermal enthalpy of the melting endotherm of about 87 J / g. As previously mentioned, the DSC values ​​obtained can vary by up to ±5°C depending on the instrument used, how the sample was prepared, and batch-to-batch variations. Generated using the DSC Q2000 module Fig.10 DSC thermogram shown. The instrument was equilibrated at 25°C and samples (1 mg to 3 mg) were heated to 230°C at a rate of 10°C / min.

[0497] Fig.13 Shown is a representative TGA thermogram of Form B. Form B exhibits less than about 0.1% weight loss upon heating from about 25°C to 110°C, confirming that Form B is an anhydrate.

[0498] Fig.14A representative GVS graph of Form B is shown. Form B exhibits a reversible moisture uptake of less than about 0.03 wt % between 0% relative humidity and 80% relative humidity at 25°C ± 0.1°C. The desorption curve shows that Form B loses moisture at a rate similar to that gained during adsorption, with limited hysteresis. No form change was observed by PXRD after the GVS experiment. Form B is non-hygroscopic (i.e., <0.2% weight gain) according to the European Pharmacopoeia (EP) classification.

[0499] Example 6: Crystallization Type 2

[0500] Type 2 is a crystalline form of racemic N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

[0501] A. (R,S)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Preparation of crystal type 2

[0502] (R,S)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide type 2 can be isolated by slurrying (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide in methanol or ethanol water mixture at high temperature for a long time or by performing several temperature cycles in chloroform, dioxane or tetrahydrofuran. These methods are shown below.

[0503] (i) Slurry at 50°C

[0504] (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide (30 mg-40 mg) Form B was slurried in a mixture of 0.25 mL of water and methanol at a volume ratio of 84:16 (methanol:water) at 50°C for 7 days or longer. The remaining solid material was isolated and dried prior to analysis. Analysis confirmed that the isolated crystalline form was Type 2.

[0505] (ii) Temperature Cycle

[0506] About 50 mg of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide amorphous was slurried in 0.3 mL of solvent (chloroform, dioxane or tetrahydrofuran). The slurry was exposed to 10 temperature cycles between 5°C and up to 3°C below the boiling point of the solvent, respectively. The temperature cycle used a ramp of 1°C / min when rising and falling. The suspension was centrifuged, the supernatant was decanted, and the solid material was dried before analysis. The analysis confirmed that the isolated crystalline form was type 2.

[0507] B. Physical Characterization of Crystalline Type 2

[0508] Characterization of Type 2 was performed using various techniques including PXRD ( Fig.15 )、Differential Scanning Calorimetry (DSC) ( Fig.16 ) and gravimetric vapor sorption (GVS) ( Fig.17 ).

[0509] Fig.15 The PXRD pattern confirmed that Type 2 is crystalline. Fig.15 The PXRD pattern of type 2 measured using reflection geometry is shown in Table 5 below. Fig.15 Selected peaks identified in the PXRD pattern.

[0510] Table 5

[0511] Peak position (°2θ) Relative strength (%) definition 8.1 4 m 10.0 100 vs 12.9 49 vs 15.6 19 s 15.8 34 vs 16.2 46 vs 17.1 52 vs 17.6 13 s 17.9 22 s 19.2 9 m 19.3 10 m 20.7 21 s 21.4 65 vs 22.0 75 vs 22.8 75 vs 23.5 20 s 24.0 39 vs 24.5 14 s 26.1 61 vs 26.5 57 vs 26.9 35 vs 29.5 9 m 30.3 12 s 30.7 11 s 31.1 8 m 31.7 9 m 32.7 5 m 34.0 7 m 37.7 13 m

[0512] Type 2 shows unique peaks at 10.0±0.2°2θ, 12.9±0.2°2θ, 17.1±0.2°2θ, 22.0±0.2°2θ, and 22.8°2θ±0.2°2θ (relative to other crystalline anhydrates). Type 2 also shows characteristic peaks at 15.8±0.2°2θ, 16.2±0.2°2θ, 26.0±0.2°2θ, 26.5±0.2°2θ, and 26.9°2θ±0.2°2θ.

[0513] Fig.16 Shown is a representative ramp DSC thermogram of Type 2. Exothermic events are plotted in the upward direction. Fig.19 The melting endotherm shown has an onset temperature of about 201°C and a heat flow of about 92 J / g for the melting endotherm. As previously mentioned, the DSC values ​​obtained can vary by as much as ±5°C depending on the instrument used, how the sample is prepared, and batch-to-batch variations.

[0514] Fig.17 A representative GVS graph for Type 2 is shown. Type 2 exhibits a reversible moisture uptake of less than about 0.5 wt % between 0% relative humidity and 80% relative humidity at 25°C ± 0.1°C. The desorption curve shows that Type 2 loses moisture at a rate similar to that gained during adsorption, with limited hysteresis. According to the European Pharmacopoeia (EP) classification, Type 2 is slightly hygroscopic (i.e., ≥ 0.2% and < 2% weight gain).

[0515] Example 7: Crystallization Solvent (Degradation / Epimerization)

[0516] Solutions of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide ("quinoline", 1.5 mg / mL) in methanol, ethanol, and acetonitrile were maintained at ambient temperature and analyzed by supercritical fluid chromatography-mass spectrometry for periods of up to 14 days. New small peaks corresponding to masses of quinoline +32 and quinoline +46 were observed in methanol and ethanol, respectively. It is believed that these peaks may come from compounds generated by adding methanol or ethanol to quinoline nitrile, namely Compound 1 and Compound 2 and their corresponding enantiomers. The structures of Compound 1 and Compound 2 are shown below:

[0517]

[0518] Small new peaks corresponding to the masses of the proposed degradants, Compound 3 and Compound 4, were also observed in methanol and ethanol. In addition, some epimerization of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide was observed. The structures of Compound 3 and Compound 4 are shown below:

[0519]

[0520] Table 6 below reports the area % data measured over time for solutions maintained in methanol.

[0521] Table 6

[0522]

[0523]

[0524] Table 7 below reports the area % data measured over time for solutions maintained in ethanol.

[0525] Table 7

[0526]

[0527] Table 8 below reports the area % data measured over time for solutions maintained in acetonitrile. Epimerization of formamide was observed in acetonitrile, but no chemical degradation.

[0528] Table 8

[0529] t0 3d 10d Quinoline enantiomers 0.87 1.34 1.73 Quinoline 99.13 98.66 98.27

[0530] Example 8: Solubility studies

[0531] A. Form B Solubility Studies

[0532] Solubility studies were performed on Form B. A small aliquot of each test solvent was added to an accurately weighed sample of Form B (about 20 mg) at ambient temperature. The aliquot volume was typically 20 μL-200 μL, until the total volume was 1.0 mL. Complete dissolution of Form B was determined by visual inspection. For solvents with a solubility of less than 20 mg / mL, the experiment was repeated using about 2 mg of Form B.

[0533] The solubility estimated in each test solvent is based on the total solvent used to provide complete dissolution. It should be noted that the actual solubility may differ to a certain extent from the estimated (i.e., visually detected) solubility due to the use of excessively large solvent aliquots, slow dissolution rates, or other factors. The estimated solubility is reported in Table 9 below.

[0534] Table 9

[0535] Solvents Solubility at 20℃(mg / mL) Acetic acid >430 acetone 20.3-22.6 Acetonitrile 20.6-22.9 Anisole <2 Butyl acetate <2 Tert-Butanol <2 Chloroform 30.9-36 Cyclohexane <2 Dichloromethane 36.2-43.4 Diisopropyl ether <2 dimethylacetamide 426-852 dimethylformamide 402-804 Dimethyl sulfoxide 424-848 Dioxane 26.6-30.4 Ethanol 2.1-2.6 Ethyl acetate <2 Heptane <2 Isopropyl acetate <2 Methanol 9.3-11.2 Methyl Ethyl Ketone 9.3-14 Methyl isobutyl ketone <2 2-Methyltetrahydrofuran <2 Methyl tert-butyl ether <2 N-Methylpyrrolidone 424-848 1-Propanol <2 Isopropyl alcohol <2 Tetrahydrofuran 24.4-27.5 Toluene <2 water <2

[0536] Suitable solvents for cooling crystallization should have high solubility for the solute and high potential recovery, i.e., the solvent should generally have high solubility for the solute at high temperatures and relatively low solubility for the solute at low temperatures (i.e., high temperature solubility coefficient). Regarding their solubility at low temperatures, solvents suitable for cooling crystallization generally have solubility in the range of about 5 mg / mL to about 20 mg / mL at 20°C.

[0537] B. FaSSIF Blank Dissolution Medium (Form A and Form B)

[0538] The solubility of Form A and Form B in phosphate buffer, pH 6.5 (FAsSIF blank), dissolution medium was determined after the crystalline form was balanced in the medium at 25°C for 1 day. High performance liquid chromatography (HPLC) was used to determine the concentration in the filtered solution. The solid residue was analyzed by PXRD to confirm the crystalline form retained. The solubility measured for Form A and Form B was 0.50 mg / ml and 0.30 mg / ml, respectively.

[0539] C. Acetonitrile solubility curve

[0540] Crystal 16 instrument is used to generate a solubility curve with temperature rise in acetonitrile. At least three accurately measured samples of (R)-N-(2-(4-cyanothiazolidine-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Form B are loaded into Crystal 16 vials, acetonitrile (1.0 mL) is loaded via a pipette, and the contents are stirred at 700 rpm. The resulting slurry is heated from 25 ° C to 70 ° C at a constant rate of 0.075 ° C / min, and the clearing point of each concentration is determined by measuring turbidity.

[0541] D. Solubility curves of acetonitrile and methanol

[0542] As the temperature rises, the solubility of Form B in acetonitrile and methanol is evaluated. (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Form B (between 14.35 mg and 74.1 mg of each sample) and solvent (1.0 mL, acetonitrile or methanol) are loaded into a Crystal 16 vial. The contents are stirred at 700 rpm, and the resulting slurry is heated from 25 ° C to 70 ° C at a constant rate of 0.075 ° C / min. The clearing point of each concentration of Form B is determined by measuring the turbidity. The solubility curve generated based on the measured data is shown in Fig.21 middle.

[0543] Example 9: Amorphous (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline- 4-Formamide

[0544] A. Amorphous (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxylate Preparation of amines

[0545] Amorphous (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide can be isolated by various methods, including freeze drying, antisolvent precipitation, flash evaporation, melt quenching, collision precipitation, and vapor diffusion. Several of these methods are shown below.

[0546] (i) Flash Distillation

[0547] (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide (25 mg-30 mg) was dissolved in hot acetone, chloroform, tetrahydrofuran or dichloromethane (400 μL-700 μL). The solution was filtered through a 0.2 μm filter directly into a heated vial on a hot plate at about 120°C, and the solvent was evaporated to give a glassy solid. The in situ generated glass residue was used without further manipulation.

[0548] (ii) Collision precipitation

[0549] A saturated solution of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide was prepared in the selected solvent and filtered through a 0.2 μm PTFE filter directly into a round bottom flask containing antisolvent (approximately 10 volumes) at ambient temperature.

[0550] (iii) Freeze drying

[0551] A solution of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide in acetonitrile / water 1:1 was frozen in liquid nitrogen and dried in a freeze dryer (Christ Alpha 2-4LDplus) overnight.

[0552] (iv) Melt quenching

[0553] (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide (20 mg-100 mg) was added to an HPLC vial and flushed with nitrogen. The vial was heated to 200° C. for five minutes and then quickly immersed in an ice / water bath to form an amorphous material.

[0554] B. Amorphous (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxylate Physical Characterization of Amines

[0555] Amorphous (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide was characterized by PXRD ( Fig.18 )、Differential Scanning Calorimetry (DSC) ( Fig.19 ) and gravimetric vapor sorption (GVS) ( Fig. 20 ) characterization. Fig.18 The PXRD pattern shows that the amorphous compound does not have a regular crystalline order.

[0556] Example 10: FAP inhibition and adhesion assays

[0557] The hFAP proteins used in the examples were either commercially available or produced in insect cells as recombinant hFAP (Gp67-6HN-TEV-FAP (M39-A757), MW 89086.7Da or cd33-FAP (27-757)-6His, MW 85926Da). Recombinant hFAP proteins were secreted from Sf21 cells in culture medium and purified by affinity chromatography (batch mode, Ni excel resin, GE Healthcare) and size exclusion chromatography (Superdex200, GE Healthcare), concentrated to 19.5 mg / mL, snap-frozen in liquid nitrogen and stored at -80°C.

[0558] A. hFAP inhibition assay

[0559] (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide was tested in a biochemical inhibition assay using hFAP enzyme (Proteros, 38-760 (PR-0071)) at 0.24nM FAC and substrate Ala-Pro-AMC (ARI-3144) at 20μM FAC. 384 low volume black plates (Greiner #784076) were used. 4μL, 0.48nM enzyme solution (100mM Tris HCl, 100mM NaCl, 0.05% Chaps, pH 7.4) was added to 40nL compound (in DMSO) in a 10CR, 3-fold dilution series from 50μM FAC. The plates were incubated in the dark at room temperature for 15 minutes. 4 μL of 40 μM substrate solution (100 mM Tris HCl, 100 mM NaCl, 0.05% Chaps, pH 7.4) was added to each well. The plate was centrifuged at 1000 rpm and incubated at room temperature in the dark for 30 minutes. The plate was read on a reader with excitation at 340 nm and emission at 460 nm. IC 50 The values ​​were determined by plotting the inhibition % versus the logarithm of the compound concentration and using a one-site dose-response model. The raw data signals were normalized using 0.5% DMSO as a 0% control and reference compound A (i.e., (S)-N-(2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)quinoline-4-carboxamide, as reported in J. Med. Chem. 2014, 57, 3053) at 50 μM as a 100% inhibitor control. The data are reported in Table 10.

[0560] B. hFAP inhibition assay (tight binder)

[0561] (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide was tested in a biochemical inhibition assay using human fibroblast activation protein alpha (hFAP) enzyme (Proteros, 38-760 (PR-0071)) at 2.4 pM FAC and substrate Ala-Pro-AMC (ARI-3144) at 20 μM FAC. 384 low volume black plates (Greiner #784076) were used. 4 μL, 4.8 pM enzyme solution (100 mM Tris HCl, 100 mM NaCl, 0.05% Chaps, pH 7.4) was added to 40 nL compound (in DMSO) in a 10CR, 3-fold dilution series from 50 nM FAC. The plates were incubated in the dark at room temperature for 15 minutes. 4 μL of 40 μM substrate solution (100 mM Tris HCl, 100 mM NaCl, 0.05% Chaps, pH 7.4) was added to each well. The plate was centrifuged at 1000 rpm and incubated at room temperature in the dark for 2.5 hours. The plate was read on a reader with excitation at 340 nm and emission at 460 nm. IC 50 The values ​​were determined by plotting the inhibition % versus the logarithm of the compound concentration and using a one-site dose-response model. The raw data signals were normalized using 0.5% DMSO as a 0% control and reference compound A (i.e., (S)-N-(2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)quinoline-4-carboxamide, as reported in J. Med. Chem. 2014, 57, 3053) at 50 μM as a 100% inhibitor control. The data are reported in Table 10.

[0562] C. hFAP binding assay

[0563] (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide was tested in a direct binding assay using an 8K surface plasmon resonance biosensor (GE Healthcare) at 20°C. Immobilization of hFAP (M39-A757) on a CMD200M sensor chip (Xantec) was performed using a standard amine coupling procedure in an immobilization buffer (10 mM HEPES, 150 mM NaCl, 0.05% Tween 20, pH 7.4). The surface was washed with 10 mM NaOH, 1 M NaCl, then activated with EDC / NHS (GE Healthcare) followed by immobilization of hFAP (in 10 mM acetate pH 5.0). Finally, the surface was inactivated with ethanolamine. The immobilization level of hFAP was approximately 4000RU-6000RU. The reference point was treated as described, omitting the injection of hFAP. Compound concentration series were injected over the immobilized protein at increasing concentrations (2 nM-500 nM) using single cycle kinetics in running buffer (20 mM TRIS, 150 mM NaCl, 0.05% Tween 20, 1% DMSO, pH 7.4). The interaction model was fully fitted to the experimental traces, enabling the determination of k on , k off and K d The data are reported in Table 10.

[0564] Table 10

[0565] Determination result <![CDATA[hFAP Inhibition Assay 1 > <h2 style=";text-align:left;direction:ltr"><![CDATA[IC <h2 style=";text-align:left;direction:ltr"> 50 <h2 style=";text-align:left;direction:ltr"> <0.37nM]]><h2 style=";text-align:left;direction:ltr"> <![CDATA[hFAP Inhibition Assay (Tight Binder) 2 > <![CDATA[IC 50 =0.084nM]]> <![CDATA[hFAP Bonding Assay 3 > <![CDATA[K d =0.012nM]]> <![CDATA[hFAP Bonding Assay 3 > <![CDATA[k (on) =4900000M -1 s -1 ]]> <![CDATA[hFAP Bonding Assay 3 > <![CDATA[k (off) =0.000046 1 / s 3 ]]>

[0566] 1 IC 50 Reported after a single measurement (n=1) or as the geometric mean of multiple measurements (n=2-3).

[0567] 2 IC 50 Reported after a single measurement (n=1) or as the geometric mean of multiple measurements (n=2-6).

[0568] 3 K d Reported after a single measurement (n=1) or as the geometric mean of multiple measurements (n=2-4). (on) and k (off) Reported after a single measurement (n=1) or as an average of multiple measurements (n=2-4). NV is invalid.

[0569] D. FAP plasma inhibition assay

[0570] Plasma (anticoagulant K2EDTA) was used as enzyme source: human plasma (pooled from AZ Biobank), mouse plasma (AZ AST Biobank), and cynomolgus monkey plasma (BioIVT,

[0571] #NHP00PLK2FNN, batch number CYN222895). Use 384-well black fluotrack PS plates (Greiner781076). 20 μL of diluted plasma (1:40 dilution of cynomolgus monkey and human plasma, 1:67 dilution of mouse plasma) in buffer (PBS, 0.1% BSA) was added to 0.6 μL of compound (in DMSO). Compounds were tested using a 10CR, 3-fold dilution series from 500 nM FAC. Two replicates were performed for each assay point on the same plate. Fluorescence blank readings were read before substrate addition. The substrate Ala-Pro-AMC (ARI-3144) stock solution (20 mM in DMSO) was diluted to a concentration of 150 μM in buffer (PBS, 0.1% BSA) and 20 μL was added to give 75 μM FAC. The plate was incubated in the dark at room temperature for 40 minutes. In a Beckman The plate was read on a reader with excitation at 360 nm and emission at 465 nm. Data were analyzed in Excel (IDBS XLfit Add-In) using a one site dose response model (4-parameter logistic fit). IC was determined by plotting the % inhibition versus the logarithm of compound concentration. 50 The raw data signals were normalized using 1.5% DMSO in diluted plasma as 0% control and 1.5% DMSO in buffer (no plasma) as 100% inhibitor control. The data are reported in Table 11.

[0572] Table 11

[0573] Determination <![CDATA[IC 50 (nM)]]> <![CDATA[FAP human plasma 1 > 0.36 <![CDATA[FAP mouse plasma 2 > 0.52 <![CDATA[FAP Cynomolgus monkey plasma 3 > 0.40

[0574] 1 IC 50 Reported after a single measurement (n=1) or as the geometric mean of multiple measurements (n=4).

[0575] 2 IC 50 Reported after a single measurement (n=1) or as the geometric mean of multiple measurements (n=4).

[0576] 3 IC 50 Reported after a single measurement (n=1).

[0577] Example 11: hPrep inhibition assay

[0578] (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide was tested in a biochemical inhibition assay using prolyl endopeptidase, prolyl oligopeptidase (hPREP) enzyme (R&D Systems, 4308-SE) at 0.6 nM FAC and substrate Z-Gly-Pro-amino-methylcoumarin (Bachem, 1-1145) at 50 μM FAC. 384 low volume black plates (Greiner #784076) were used. 4 μL, 1.2 nM enzyme solution (25 mM Tris HCl, 250 mM NaCl, 0.01% Triton X-100, 5 mM glutathione, pH 7.5) was added to 40 nL compound (in DMSO) in a 10CR, 3-fold dilution series from 50 μM FAC. The plate was incubated at room temperature in the dark for 15 minutes. 4 μL of 100 μM substrate solution (25 mM Tris HCl, 250 mM NaCl, 0.01% Triton X-100, 5 mM glutathione, pH 7.5) was added to each well. The plate was centrifuged at 1000 rpm and incubated at room temperature in the dark for 20 minutes. The plate was read on a reader with excitation at 340 nm and emission at 460 nm. IC 50 The values ​​were determined by plotting the inhibition % versus the logarithm of the compound concentration and using a one site dose response model. The raw data signals were normalized using 0.5% DMSO as a 0% control and reference compound B (i.e., (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-7-methylquinoline-4-carboxamide) at 50 μM as a 100% inhibitor control. The data are reported in Table 12.

[0579] Table 12

[0580]

[0581] 1 IC 50 Reported after a single measurement (n=1) or as the geometric mean of multiple measurements (n=2-9).

[0582] 2 Reference compound B: (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-7-methylquinoline-4-carboxamide.

[0583] 3 Reference compound C: (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-quinoline-4-carboxamide.

[0584] Example 12: hDPP inhibition assay

[0585] A. hDPP7 inhibition assay

[0586] (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide was tested in a biochemical inhibition assay using human dipeptidyl peptidase 7 (hDPP7) enzyme (BPS Bioscience, #80070) at 15 nM FAC and substrate Ala-Pro-amino-methylcoumarin (BPS Bioscience, #80305) at 5 μM FAC. Enzymatic reactions were performed in duplicate in 50 μL DPP assay buffer (BPS Bioscience, #80300) at room temperature for 30 minutes. Compound solutions (in DMSO) were prepared in 10CR, 3-fold dilution series in assay buffer at ten times higher than the final concentration, and 5 μL of the dilutions were added to 50 μL of the reaction, resulting in a maximum compound concentration of 100 μM FAC and a 1% DMSO concentration in all wells. Plates were read on a Tecan Infinite M1000 microplate reader at excitation 340 nm and emission 460 nm. Data were analyzed in Graph Pad Prism. IC 50 Values ​​were determined by plotting the % inhibition versus the logarithm of compound concentration and using a one site dose response model. Raw data signals were normalized using 1% DMSO as the 0% control and no enzyme as the 100% inhibitor control. Data are reported in Table 13.

[0587] B. hDPP8 inhibition assay

[0588] (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide was tested in a biochemical inhibition assay using human dipeptidyl peptidase 8 (hDPP8) enzyme (BPS Bioscience, #80080) at 1.5 nM FAC and substrate Ala-Pro-amino-methylcoumarin (BPS Bioscience, #80305) at 5 μM FAC. Enzymatic reactions were performed in duplicate in 50 μL DPP assay buffer (BPS Bioscience, #80300) at room temperature for 30 minutes. Compound solutions (in DMSO) were prepared in 10CR, 3-fold dilution series in assay buffer at ten times higher than the final concentration, and 5 μL of the dilutions were added to 50 μL of the reaction, resulting in a maximum compound concentration of 100 μM FAC and a 1% DMSO concentration in all wells. Plates were read on a Tecan Infinite M1000 microplate reader at excitation 340 nm and emission 460 nm. Data were analyzed in Graph Pad Prism. IC 50 Values ​​were determined by plotting the % inhibition versus the logarithm of compound concentration and using a one site dose response model. Raw data signals were normalized using 1% DMSO as the 0% control and no enzyme as the 100% inhibitor control. Data are reported in Table 13.

[0589] C. hDPP9 inhibition assay

[0590] (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide was tested in a biochemical inhibition assay using human dipeptidyl peptidase 9 (hDPP9) enzyme (BPS Bioscience, #80090) at 0.4 nM FAC and substrate Ala-Pro-amino-methylcoumarin (BPS Bioscience, #80305) at 5 μM FAC. Enzymatic reactions were performed in duplicate in 50 μL DPP assay buffer (BPS Bioscience, #80300) at room temperature for 30 minutes. Compound solutions (in DMSO) were prepared in 10CR, 3-fold dilution series in assay buffer ten times higher than the final concentration, and 5 μL of the dilutions were added to 50 μL of the reaction, resulting in a maximum compound concentration of 100 μM FAC and a 1% DMSO concentration in all wells. Plates were read on a Tecan Infinite M1000 microplate reader at excitation 340 nm and emission 460 nm. Data were analyzed in Graph Pad Prism. IC 50Values ​​were determined by plotting the % inhibition versus the logarithm of compound concentration and using a one site dose response model. Raw data signals were normalized using 1% DMSO as the 0% control and no enzyme as the 100% inhibitor control. Data are reported in Table 13.

[0591] Table 13

[0592] Determination <![CDATA[IC 50 (nM) 1 ]]> hDPP7 33000 hDPP8 13000 hDPP9 1900

[0593] 1 IC 50 Reported after a single measurement (n=1).

[0594] ***

[0595] Although specific embodiments and examples have been described above, these embodiments and examples are merely illustrative and do not limit the scope of the present disclosure. Changes and modifications may be made according to one of ordinary skill in the art without departing from the disclosure of the broader aspects as defined in the following claims. For example, any embodiment described herein may be combined with any other suitable embodiment described herein to provide additional embodiments.

Claims

1. A crystalline form of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

2. The crystalline form according to claim 1, which is a crystalline form of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

3. The crystalline form of claim 2, characterized by a transmission X-ray powder diffraction pattern comprising peaks at 18.7 ± 0.2° 2θ and 22.4° 2θ ± 0.2° 2θ.

4. The crystalline form of claim 3, wherein the transmission X-ray powder diffraction pattern further comprises at least one peak selected from the group consisting of 12.0±0.2°2θ, 18.4±0.2°2θ, 19.8±0.2°2θ, and 21.7°2θ±0.2°2θ.

5. The crystalline form according to any one of claims 2 to 4, further characterized by: solid state 13 C NMR spectrum comprising at least one peak selected from the group consisting of 166.9±0.2 ppm, 130.2±0.2 ppm, 118.6±0.2 ppm, 117.4±0.2 ppm, 110.0±0.2 ppm, 49.9±0.2 ppm, 46.6±0.2 ppm, and 34.5±0.2 ppm.

6. The crystalline form of claim 5, wherein the solid 13 The C NMR spectrum contained peaks at 49.9±0.2 ppm and 46.6±0.2 ppm.

7. The crystalline form of any one of claims 2 to 6, further characterized by a differential scanning calorimetry curve comprising a melting endotherm with an onset between about 185°C and about 200°C.

8. The crystalline form of any one of claims 3 to 7, further characterized by a thermogravimetric analysis thermogram, wherein the crystalline form exhibits a weight loss of less than about 0.5 wt% from about 25°C to about 110°C.

9. The crystalline form of any one of claims 3 to 8, further characterized by a gravimetric vapor sorption plot, wherein the crystalline form exhibits a reversible moisture uptake of less than about 0.5 wt % from about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C.

10. The crystalline form according to any one of claims 3 to 9, wherein the crystalline form is a crystalline anhydrate.

11. The crystalline form of claim 2, wherein: The transmission X-ray powder diffraction pattern comprises a 2θ selected from 12.0±0.2°, at least one peak from the group consisting of 18.4±0.2°2θ, 19.8±0.2°2θ, and 21.7°2θ±0.2°2θ; and The transmission X-ray powder diffraction pattern does not include peaks with moderate or stronger relative intensities at 18.7±0.2°2θ and 22.4°2θ±0.2°2θ.

12. The crystalline form of claim 11, wherein the transmission X-ray powder diffraction pattern further comprises at least two peaks selected from the group consisting of 12.0±0.2°2θ, 18.4±0.2°2θ, 19.8±0.2°2θ, and 21.7°2θ±0.2°2θ.

13. The crystalline form of any one of claims 2, 11 or 12, further characterized by: solid state 13 C NMR spectrum comprising at least one peak selected from the group consisting of 166.1±0.2 ppm, 130.7±0.2 ppm, 117.9±0.2 ppm, 108.6±0.2 ppm, and 35.2±0.2 ppm.

14. The crystalline form of claim 13, wherein the solid 13 The C NMR spectrum contained peaks at 166.1±0.2 ppm, 117.9±0.2 ppm, and 108.6±0.2 ppm.

15. The crystalline form of claim 2 or any one of claims 11 to 14, further characterized by a differential scanning calorimetry curve comprising a melting endotherm with an onset temperature between about 165°C and about 180°C.

16. The crystalline form of any one of claims 11 to 15, further characterized by a thermogravimetric analysis thermogram, wherein the crystalline form exhibits a weight loss of less than about 0.5 wt% from about 25°C to about 110°C.

17. The crystalline form of any one of claims 11 to 16, further characterized by a gravimetric vapor sorption plot, wherein the crystalline form exhibits a reversible moisture uptake of less than about 0.5 wt % from about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C.

18. The crystalline form according to any one of claims 11 to 17, wherein the crystalline form is a crystalline anhydrate.

19. The crystalline form of claim 1, which is a crystalline form of (R,S)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide.

20. A composition comprising at least two crystalline forms selected from the group consisting of: The crystalline form according to any one of claims 3 to 10; The crystalline form according to any one of claims 11 to 18; and The crystalline form according to claim 19.

21. A pharmaceutical composition comprising the crystalline form according to any one of claims 1 to 19, and one or more pharmaceutically acceptable excipients.

22. A method of treating or preventing a FAP-mediated disorder in a subject suffering from or susceptible to a FAP-mediated disorder, the method comprising administering to the subject a therapeutically effective amount of a crystalline form according to any one of claims 1 to 19.

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