Solid state forms of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholino-quinoline-4-carboxamide

Abstract

Solid state forms of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-4-carboxamide; corresponding pharmaceutical compositions; use for treating or preventing prolyl endopeptidase fibroblast activation protein (FAP)-mediated conditions; kits; and methods of preparation.

Description

Technical Field

[0001] (Cross - Reference to Related Applications) 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 application is incorporated by reference in its entirety for all purposes.

[0002] The present disclosure generally relates to solid - state forms of N-(2-(4 - cyanothiazolidin - 3 - yl)-2 - oxoethyl)-6 - morpholinquinoline - 4 - carboxamide, including crystalline forms of (R)-N-(2-(4 - cyanothiazolidin - 3 - yl)-2 - oxoethyl)-6 - morpholinquinoline - 4 - carboxamide. The present disclosure further relates to pharmaceutical compositions comprising crystalline forms of (R)-N-(2-(4 - cyanothiazolidin - 3 - yl)-2 - oxoethyl)-6 - morpholinquinoline - 4 - carboxamide; the use of pharmaceutical compositions comprising crystalline forms of (R)-N-(2-(4 - cyanothiazolidin - 3 - yl)-2 - oxoethyl)-6 - morpholinquinoline - 4 - carboxamide for treating or preventing prolyl endopeptidase fibroblast activation protein (FAP)-mediated conditions; kits comprising pharmaceutical compositions comprising crystalline forms of (R)-N-(2-(4 - cyanothiazolidin - 3 - yl)-2 - oxoethyl)-6 - morpholinquinoline - 4 - carboxamide; and methods for preparing crystalline forms of (R)-N-(2-(4 - cyanothiazolidin - 3 - yl)-2 - oxoethyl)-6 - morpholinquinoline - 4 - carboxamide.

Background Art

[0003] FAP, a 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, which are a major aspect of NASH predicting disease outcome (Hepatology 1999, 29, 1768), and involved in fibrosis formation (Gastroenterology 2020, 158, 1611). FAP can also exist as a shed plasma protease. An increase in circulating FAP levels is associated with the severity of NASH disease (Diabetes Res Clin Pract 2015, 108, 466).

[0004] FAP has a consensus cleavage motif following Gly-Pro and exhibits both endopeptidase and exopeptidase activities. Known enzymatic activities include the cleavage of collagen (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) on the cell surface of activated fibroblasts generates a fibrosis-promoting environment. FAP cleavage of α2AP results in more efficient cross-linking of α2AP to fibrin, leading to a decrease in fibrin clearance. FAP cleavage of FGF21 inactivates the metabolic effects of FGF21 (Biochem J 2016, 473, 605). All of these activities are associated with the worsening of NASH disease, and inhibiting FAP has the potential to treat NASH and other conditions by affecting multiple mechanisms.

[0005] Inhibition of FAP activity is an untapped therapeutic approach for treating NASH and other diseases associated with such activity. Approved pharmacological agents that generally inhibit FAP activity or specifically inhibit FAP activity are not currently available. Accordingly, there is a need for FAP inhibitors, particularly those having suitable pharmacological selectivity and bioavailability and physical properties suitable for the manufacture of the prodrug and the formulation of the corresponding drug product.

[0006] The present disclosure addresses this large unmet need by providing solid state forms of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide that are suitable for use in pharmaceutical compositions and methods for the treatment of FAP-mediated conditions such as NASH. SUMMARY OF THE INVENTION

[0007] In one aspect, the present disclosure provides a solid state form of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide.

[0008] In another aspect, the present disclosure provides a crystalline form of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide.

[0009] In another aspect, the present disclosure provides crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide Form A.

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

[0011] In another aspect, the present disclosure provides a crystalline form of (R,S)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide.

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

[0013] 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.

[0014] In another aspect, the present disclosure provides a method of treating or preventing a prolyl endopeptidase fibroblast activation protein (FAP)-mediated condition, the method comprising 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.

[0015] In another aspect, the present disclosure provides the 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 prolyl endopeptidase fibroblast activation protein (FAP)-mediated condition.

[0016] In another aspect, the present disclosure provides the use of a crystalline form of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide in the manufacture of a medicament for treating or preventing a prolyl endopeptidase fibroblast activation protein (FAP)-mediated condition.

[0017] 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-morpholinquinoline-4-carboxamide.

[0018] In another aspect, the present disclosure provides a method for preparing a crystalline form of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide. BRIEF DESCRIPTION OF THE DRAWINGS

[0019]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Figure 7

Figure 8

Figure 9

Figure 10

Figure 11

Figure 12

Figure 13

Figure 14

Figure 15

Figure 16

Figure 17

Figure 18

Figure 19

Figure 20

Figure 21

Best Mode for Carrying Out the Invention

[0020] Numerous embodiments are detailed throughout this specification and will be apparent 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. Various alternatives to the described embodiments may be used in practicing the invention.

[0021] I. Definitions With respect to the embodiments disclosed herein, the following terms have the meanings set forth below.

[0022] References to "a" or "an" mean "one or more". Throughout, the plural and singular should be treated as interchangeable unless otherwise indicated by the number of the reference.

[0023] When ranges are used herein, for example, to describe physical or chemical properties, all combinations and sub - combinations of the range and specific embodiments therein are intended to be included. The use of the term "about" or "approximately" when referring to a number or numerical range means that the number or numerical range so referred to is an approximation within experimental variability (or within statistical experimental error), and thus the number or numerical range may vary, for example, from 1% to 15% of the recited number or numerical range.

[0024] Unless the context requires otherwise, the words "comprise", "comprises" or "comprising" should be construed inclusively rather than exclusively, and based on and for the clear understanding that the Applicant intends that each of these words should be so construed when interpreting this patent including the following claims.

[0025] The term "amorphous form" refers to a form of a compound lacking long - range crystalline order.

[0026] As used herein, the terms “co-administered,” “co-administering,” “administered in combination with,” and “administering in combination with” include 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.

[0027] The term “crystalline form” is intended to include all crystalline forms of a compound, including, for example, polymorphs, pseudopolymorphs, solvates, hydrates, non-solvated polymorphs (including anhydrates), and conformational polymorphs, as well as mixtures thereof, unless a particular crystalline form is recited.

[0028] The term “therapeutically effective amount” of a pharmacological agent is an amount sufficient to produce a beneficial or desired result, including clinical results, and thus varies depending on the circumstances in which it is administered. For example, when an agent is administered to treat a liver disease, the therapeutically effective amount of the pharmacological agent is an amount of the agent sufficient to provide an anti-liver disease effect in a subject as compared to the outcome obtained without administration of the agent, either alone or in combination with additional therapies.

[0029] The term “pharmaceutically acceptable” is used adjectivally herein to mean that the modified noun is suitable for use as a pharmaceutical or part of a pharmaceutical. For example, the terms “pharmaceutical carrier” or “pharmaceutically acceptable excipient” are intended to include any and all carriers or excipients suitable for use in mammals, particularly humans.

[0030] The term “reflection” or “reflection mode,” when used in conjunction with powder X-ray diffraction, refers to the reflection (also known as the Bragg-Brentano) sampling mode.

[0031] The term "prevent" is readily understood by a normally skilled physician and is intended to have its ordinary meaning with respect to the treatment of a particular condition, including primary prevention to prevent the onset of the condition, and secondary prevention where the condition has already occurred and the patient is protected temporarily or permanently from disease progression or worsening or the development of new symptoms associated with the condition.

[0032] The term "solvate" refers to the crystalline phase of a compound that is physically associated with one or more molecules of a solvent. The crystalline phase of a compound that is physically associated with one or more molecules of water is called a "hydrate".

[0033] The term "transmission" or "transmission mode", when used in conjunction with powder X-ray diffraction, refers to the transmission (also known as the Debye-Scherrer) sampling mode.

[0034] The term "treat" is readily understood by a normally skilled physician and is intended to have its ordinary meaning with respect to the treatment of a particular condition, and may include (1) reducing the degree or cause of the condition being treated, and / or (2) alleviating or improving one or more symptoms associated with the condition. Treatment of a liver disease may include, for example, stabilizing (i.e., not worsening), delaying, or retarding the spread or progression of the liver disease; extending the survival period compared to what would be expected in the absence of treatment; and / or improving or alleviating, in whole or in part, the severity of the liver disease.

[0035] As used herein, "enantiomeric purity" refers to the relative amount of the presence of a particular enantiomer expressed as a percentage relative to the other enantiomer. For example, when a compound that can potentially have an (R)- or (S)-isomer configuration is present as a racemic mixture, the enantiomeric purity is about 50% with respect to either the (R)- or (S)-isomer. When the compound has one enantiomeric form that is more dominant than the other, for example, 80% of the (S)-isomer and 20% of the (R)-isomer, the enantiomeric purity with respect to the (S)-enantiomeric form of the compound is 80%. The enantiomeric purity of a compound can be determined by several methods, including chromatography using a chiral support, polarimetry of the rotation of polarized light, nuclear magnetic resonance spectroscopy using a chiral shift reagent including but not limited to a lanthanide-containing chiral complex or a Pirkle reagent, or derivatization of the compound using a chiral compound such as Mosher acid followed by chromatography or nuclear magnetic resonance spectroscopy, including but not limited to these.

[0036] In some embodiments, an enantiomerically enriched composition has a higher potency with respect to the therapeutic utility per unit mass than the racemic mixture of the composition. Enantiomers can be isolated from the mixture by methods known to those skilled in the art, including chiral high performance liquid chromatography (HPLC) and the formation and crystallization of chiral salts. Alternatively, the preferred enantiomer 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.

[0037] The terms "enantiomerically enriched" and "non-racemic", as used herein, refer to a composition in which the weight percentage of one enantiomer is greater than the amount of that one enantiomer in the control mixture of the racemic composition (e.g., greater than 1:1 on a weight basis). For example, an enantiomerically enriched preparation of the (R)-enantiomer means a preparation of a compound having greater than 50% by weight of the (R)-enantiomer, e.g., at least 75% by weight, or e.g., at least 80% by weight, relative to the (S)-enantiomer. In some embodiments, the enrichment can significantly exceed 80% by weight, providing a "substantially enantiomerically enriched" or "substantially non-racemic" preparation, which refers to a preparation of a composition having at least 85% by weight, e.g., at least 90% by weight, or e.g., at least 95% by weight of one enantiomer relative to the other enantiomer. The terms "enantiomerically pure" or "substantially enantiomerically pure" refer to a composition containing at least 98% of a single enantiomer and less than 2% of the opposite enantiomer.

[0038] II. Crystal Forms of N-(2-(4-Cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-4-carboxamide The compound, which is an active pharmaceutical ingredient in a pharmaceutical product, can potentially exist in different solid state forms exhibiting different physical properties. These differences in physical properties can affect the manufacture and formulation of the pharmaceutical product. Such physical properties can include, but are not limited to, (1) packing 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 stability to moisture and stability under storage conditions), (4) surface properties such as surface area, wettability, interfacial tension, and shape, (5) mechanical properties such as hardness, tensile strength, compressibility, formability, handleability, flowability, and blend, and (6) filtration properties. Therefore, a solid state form of the compound, particularly a crystalline form of the compound, that provides an improvement in one or more of these physical properties compared to other solid state forms of the compound is desirable. Thus, the discovery of new solid state forms of pharmaceutically useful compounds provides potential opportunities to improve the performance characteristics of the corresponding pharmaceutical products and the related manufacturing processes.

[0039] The present disclosure provides solid state forms of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-4-carboxamide. In one aspect, the solid state form is a crystalline form. Each crystalline form described herein has one or more of the above advantageous properties compared to one or more of the other solid state forms of the compound. In another aspect, the solid state form is a crystalline anhydrate. In a further aspect, the crystalline form is substantially pure. As used herein, 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 state 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.

[0040] In some embodiments, the present disclosure provides a crystalline form of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-4-carboxamide having the following chemical structure.

[0041]

Chemical formula

[0042] In some embodiments, the present disclosure provides crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-4-carboxamide Form A.

[0043] In some embodiments, Form A is characterized by a transmission X-ray powder diffraction pattern that (i) includes 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°, and (ii) does not include peaks with relative intensities above medium intensity at 18.7 ± 0.2° 2θ and 22.4° 2θ ± 0.2°. In one aspect, the transmission X-ray powder diffraction pattern does not include peaks at 18.7 ± 0.2° 2θ and 22.4° 2θ ± 0.2°. In another aspect, the transmission X-ray powder diffraction pattern includes at least 2, 3, or 4 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°. In another aspect, the transmission X-ray powder diffraction pattern includes peaks at 12.0 ± 0.2° 2θ, 18.4 ± 0.2° 2θ, 19.8 ± 0.2° 2θ, and 21.7° 2θ ± 0.2°. In another aspect, the transmission X-ray powder diffraction pattern further includes at least 1, 2, 3, or 4 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°. In another aspect, the transmission X-ray powder diffraction pattern further includes peaks at 13.1 ± 0.2° 2θ, 14.4 ± 0.2° 2θ, 17.5 ± 0.2° 2θ, and 21.1° 2θ ± 0.2°. In another aspect, the transmission X-ray powder diffraction pattern includes 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°. In another aspect, the transmission X-ray powder diffraction pattern is substantially the same as the transmission X-ray powder diffraction pattern of FIG. 1.

[0044] In some embodiments, Form A is characterized by a solid state NMR spectrum that includes 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 state 13 13C NMR spectrum includes peaks at 166.1 ± 0.2 ppm, 117.9 ± 0.2 ppm and 108.6 ± 0.2 ppm. In another aspect, the solid state 13 13C NMR spectrum includes 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 state 13 13C NMR spectrum includes 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 state 13 13C 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. In another aspect, the solid state 13 13C 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. In another aspect, the solid state 13 13C NMR spectrum does not include peaks at 49.9 ± 0.2 ppm and 46.6 ± 0.2 ppm. In another aspect, the solid state 13 13C NMR spectrum does not include peaks at 49.9 ± 0.2 ppm and 46.6 ± 0.2 ppm. In another aspect, the solid state 13The 13C 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, in the solid state 13 The 13C 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.

[0045] In some embodiments, Form A is characterized by a differential scanning calorimetry curve that includes an endotherm with an onset temperature of about 165°C to about 180°C. In one aspect, the endotherm includes an endotherm with an onset temperature of about 165°C to about 177°C. In another aspect, the endotherm starts at 171°C ± 5°C. In another aspect, the differential scanning calorimetry curve is substantially the same as the differential scanning calorimetry curve of FIG. 6.

[0046] In some embodiments, Form A is further characterized by a thermogravimetric analysis thermogram in which the crystalline form exhibits a weight loss of less than about 0.5 wt% at 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 substantially the same as the thermogravimetric analysis thermogram of FIG. 7.

[0047] In some embodiments, Form A is characterized by a weight vapor sorption plot in which the crystalline form exhibits a reversible moisture uptake of less than about 0.5 wt% at 25°C ± 0.1°C and from about 0% relative humidity to about 80% relative humidity. 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 weight vapor sorption plot is substantially the same as the weight vapor sorption plot of FIG. 8.

[0048] In some embodiments, Form A has the above physical characterization embodiments (transmission X-ray powder diffraction, solid state 13characterized by at least two of (13C NMR, differential scanning calorimetry, thermogravimetric analysis, and / or gravimetric vapor sorption).

[0049] In some embodiments, Form A is the above physical characterization embodiments (powder X-ray diffraction, solid state 13 characterized by at least three of 13C NMR, differential scanning calorimetry, thermogravimetric analysis, and / or gravimetric vapor sorption).

[0050] In some embodiments, Form A is the above physical characterization embodiments (powder X-ray diffraction, solid state 13 characterized by at least four of 13C NMR, differential scanning calorimetry, thermogravimetric analysis, and / or gravimetric vapor sorption).

[0051] In some embodiments, Form A is characterized by: a powder X-ray 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 not comprising peaks with relative intensities above medium intensity at 18.7 ± 0.2° 2θ and 22.4° 2θ ± 0.2° 2θ; a 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; and 13 a differential scanning calorimetry curve comprising an endotherm for melting having an onset temperature of about 165°C to about 180°C.

[0052] In some embodiments, Form A is characterized by: a powder X-ray 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 not comprising peaks with relative intensities above medium intensity at 18.7 ± 0.2° 2θ and 22.4° 2θ ± 0.2° 2θ; ​A solid state including 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 13 13C NMR spectrum; A differential scanning calorimetry curve including an endothermic melting peak having an onset temperature of about 165°C to about 177°C; and A thermogravimetric analysis thermogram in which the crystalline form shows a weight loss of less than about 0.2 wt% at about 25°C to about 110°C.

[0053] In some embodiments, Form A is characterized by: An X-ray powder diffraction pattern including 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°, and not including peaks with relative intensities above medium intensity at 18.7 ± 0.2° 2θ and 22.4° 2θ ± 0.2°; A solid state including 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 13 13C NMR spectrum; A differential scanning calorimetry curve including an endothermic melting peak having an onset temperature of about 165°C to about 177°C; and A gravimetric vapor sorption plot in which the crystalline form shows a reversible moisture uptake of less than about 0.2 wt% at about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C.

[0054] In some embodiments, Form A is characterized by: An X-ray powder diffraction pattern including 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°, and not including peaks at 18.7 ± 0.2° 2θ and 22.4° 2θ ± 0.2° with relative intensities above medium intensity; A solid state including 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 ppm13 13C NMR spectrum; Differential scanning calorimetry curve including an endothermic melting starting at 171 °C ± 5 °C; Thermogravimetric analysis thermogram in which the crystalline form shows a weight loss of less than about 0.2 wt% at about 25 °C to about 110 °C; and Weight vapor sorption plot in which the crystalline form shows a reversible water uptake of less than about 0.2 wt% at about 0% relative humidity to about 80% relative humidity at 25 °C ± 0.1 °C.

[0055] In some embodiments, Form A is a crystalline anhydrate.

[0056] In some embodiments, Form A has a long needle-like form.

[0057] In some embodiments, Form A is substantially free of any other crystalline forms of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-4-carboxamide.

[0058] In some embodiments, Form A contains less than 5 wt% of any other crystalline forms of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-4-carboxamide.

[0059] In some embodiments, Form A contains less than 10 wt% of any other crystalline forms of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-4-carboxamide.

[0060] In some embodiments, the present disclosure provides crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-4-carboxamide Form B.

[0061] In some embodiments, Form B is characterized by a transmission X-ray powder diffraction pattern that includes 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 a relative intensity of at least medium intensity or greater. In another aspect, the transmission X-ray powder diffraction pattern further includes at least 1, 2, 3, or 4 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 further includes 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 includes at least 1, 2, 3, or 4 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 includes 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 includes peaks 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, 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 is substantially the same as the transmission X-ray powder diffraction pattern of FIG. 9.

[0062] In some embodiments, Form B comprises 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 a solid state 13 and is further characterized by a solid-state 13 13C NMR spectrum. In one aspect, the solid-state 13 13C NMR spectrum comprises peaks at 49.9±0.2 ppm and 46.6±0.2 ppm. In another aspect, the solid-state 13 13C 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. In another aspect, the solid-state 13 13C 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-state 13 13C 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-state 13The 13C 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. In another aspect, in the solid state 13 The 13C NMR spectrum does not include peaks at 166.1 ± 0.2 ppm, 117.9 ± 0.2 ppm, and 108.6 ± 0.2 ppm. In another aspect, in the solid state 13 The 13C NMR spectrum does not include 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.

[0063] In some embodiments, Form B is characterized by a differential scanning calorimetry curve that includes an endothermic melting starting at about 185 °C to 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 substantially the same as the differential scanning calorimetry curve of FIG. 12.

[0064] In some embodiments, Form B is further characterized by a thermogravimetric analysis thermogram in which the crystalline form exhibits a weight loss of less than about 0.5 wt% at 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 substantially the same as the thermogravimetric analysis thermogram of FIG. 13.

[0065] In some embodiments, Form B is characterized by a weight vapor sorption plot in which the crystalline form exhibits a reversible moisture uptake of less than about 0.5 wt% at 25 °C ± 0.1 °C from about 0% relative humidity to about 80% relative humidity. 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 weight vapor sorption plot is substantially the same as the weight vapor sorption plot of FIG. 14.

[0066] In some embodiments, Form B is characterized by at least two of the above physical characterization embodiments (transmission X-ray powder diffraction, solid state 13 C NMR, differential scanning calorimetry, thermogravimetric analysis, and / or gravimetric vapor sorption).

[0067] In some embodiments, Form B is characterized by at least three of the above physical characterization embodiments (transmission X-ray powder diffraction, solid state 13 C NMR, differential scanning calorimetry, thermogravimetric analysis, and / or gravimetric vapor sorption).

[0068] In some embodiments, Form B is characterized by at least four of the above physical characterization embodiments (transmission X-ray powder diffraction, solid state 13 C NMR, differential scanning calorimetry, thermogravimetric analysis, and / or gravimetric vapor sorption).

[0069] In some embodiments, Form B is characterized by: A transmission X-ray powder diffraction pattern including peaks at 18.7 ± 0.2° 2θ and 22.4° 2θ ± 0.2° 2θ; 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 in solid state 13 C NMR spectrum; and A differential scanning calorimetry curve including an endothermic melting starting at about 185°C to about 200°C.

[0070] In some embodiments, Form B is characterized by: A transmission X-ray powder diffraction pattern including peaks at 18.7 ± 0.2° 2θ and 22.4° 2θ ± 0.2° 2θ; 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 solid state13 13C NMR spectrum; Differential scanning calorimetry curve including an endothermic melting starting at about 185 °C to about 200 °C; and Thermogravimetric analysis thermogram in which the crystalline form shows a weight loss of less than about 0.2 wt% at about 25 °C to about 110 °C.

[0071] In some embodiments, Form B is characterized by: A transmission X-ray powder diffraction pattern including peaks at 18.7 ± 0.2° 2θ and 22.4° 2θ ± 0.2° 2θ; Solid state including 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 13 13C NMR spectrum; Differential scanning calorimetry curve including an endothermic melting starting at about 185 °C to about 200 °C; and A weight vapor sorption plot in which the crystalline form shows a reversible water uptake of less than about 0.2 wt% at about 0% relative humidity to about 80% relative humidity at 25 °C ± 0.1 °C.

[0072] In some embodiments, Form B is characterized by: A transmission X-ray powder diffraction pattern including peaks at 18.7 ± 0.2° 2θ and 22.4° 2θ ± 0.2° 2θ; Solid state including 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 13 13C NMR spectrum; Differential scanning calorimetry curve including an endothermic melting starting at 191 °C ± 5 °C; Thermogravimetric analysis thermogram in which the crystalline form shows a weight loss of less than about 0.1 wt% at about 25 °C to about 110 °C; and A weight vapor sorption plot in which the crystalline form shows a reversible water uptake of less than about 0.1 wt% at about 0% relative humidity to about 80% relative humidity at 25 °C ± 0.1 °C.

[0073] In some embodiments, Form B is a crystalline anhydrate.

[0074] In some embodiments, Form B has an elongated needle-like morphology.

[0075] In some embodiments, Form B is substantially free of any other crystalline forms of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-4-carboxamide.

[0076] 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-morpholin-4-ylquinoline-4-carboxamide.

[0077] 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-morpholin-4-ylquinoline-4-carboxamide.

[0078] In some embodiments, the present disclosure provides a crystalline form of (R,S)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-4-carboxamide (i.e., a crystalline form of racemic N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholino-quinoline-4-carboxamide).

[0079] In some embodiments, the present disclosure provides crystalline (R,S)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-4-carboxamide Form 2.

[0080] In some embodiments, the Type 2 is characterized by a reflected 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°. In one aspect, the reflected 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°. In another aspect, the reflected X-ray powder diffraction pattern further comprises at least 1, 2, 3, 4, or 5 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°. In another aspect, the reflected 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°. In another aspect, the reflected X-ray powder diffraction pattern comprises peaks at 8.1 ± 0.2° 2θ, 10.0 ± 0.2° 2θ, 12.9 ± 0.2° 2θ, 15.6 ± 0.2° 2θ, 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, 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, 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, 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, 31.1 ± 0.2° 2θ, 31.7 ± 0.2° 2θ, 32.7 ± 0.2° 2θ, 34.0 ± 0.2° 2θ, and 37.7 ± 0.2°. In another aspect, the reflected X-ray powder diffraction pattern is substantially the same as the reflected X-ray powder diffraction pattern of FIG. 15.

[0081] In some embodiments, Form 2 is characterized by a differential scanning calorimetry curve that includes an endotherm from about 195 °C to about 210 °C. In one aspect, the endotherm has an onset temperature from about 195 °C to about 207 °C. In another aspect, the endotherm has an onset temperature of 201 °C ± 5 °C. In another aspect, the differential scanning calorimetry curve is substantially the same as the differential scanning calorimetry curve of FIG. 16.

[0082] In some embodiments, Form 2 is characterized by a gravimetric vapor sorption plot in which the crystalline form exhibits reversible water uptake of less than about 1.0 wt% at about 0% relative humidity to about 80% relative humidity at 25 °C ± 0.1 °C. In one aspect, the reversible water uptake is less than about 0.7 wt%. In another aspect, the reversible water uptake is about 0.5 wt%. In another aspect, the gravimetric vapor sorption plot is substantially the same as the gravimetric vapor sorption plot of FIG. 17.

[0083] In some embodiments, Form 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).

[0084] In some embodiments, Form 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).

[0085] In some embodiments, Form 2 is characterized by: a reflection X-ray powder diffraction pattern that includes 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; a differential scanning calorimetry curve that includes an endotherm with an onset temperature from about 190 °C to about 210 °C; and a gravimetric vapor sorption plot in which the crystalline form exhibits reversible water uptake of less than about 1.0 wt% at about 0% relative humidity to about 80% relative humidity at 25 °C ± 0.1 °C.

[0086] In some embodiments, Form 2 is characterized by: a powder X-ray diffraction pattern that includes 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°; a differential scanning calorimetry curve that includes an endotherm for melting having an onset temperature of about 195°C to about 207°C; and a gravimetric vapor sorption plot in which the crystalline form exhibits less than about 0.7 weight % reversible moisture uptake at 25°C ± 0.1°C and from about 0% relative humidity to about 80% relative humidity.

[0087] In some embodiments, Form 2 is characterized by: a powder X-ray diffraction pattern that includes 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°; a differential scanning calorimetry curve that includes an endotherm for melting that begins at 201°C ± 5°C; and a gravimetric vapor sorption plot in which the crystalline form exhibits about 0.5 weight % reversible moisture uptake at 25°C ± 0.1°C and from about 0% relative humidity to about 80% relative humidity.

[0088] In some embodiments, Form 2 is a crystalline anhydrate.

[0089] In some embodiments, Form 2 is substantially free of any other crystalline forms of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-4-carboxamide.

[0090] In some embodiments, Form 2 includes less than 5 weight % of any other crystalline forms of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-4-carboxamide.

[0091] In some embodiments, Form 2 comprises less than 10% by weight of any other crystalline form of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-4-carboxamide.

[0092] 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 Form 2.

[0093] 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.

[0094] Exemplary embodiments: Embodiment 1: A crystalline form of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-4-carboxamide.

[0095] Embodiment 2: The crystalline form according to Embodiment 1, which is a crystalline form of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-4-carboxamide.

[0096] Embodiment 3: The crystalline form according to Embodiment 2, which 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.

[0097] Embodiment 4: The crystalline form according to Embodiment 3, wherein the peaks at 18.7±0.2° 2θ and 22.4° 2θ±0.2° 2θ have a relative intensity of at least medium intensity or more.

[0098] Embodiment 5: The crystalline form according to Embodiment 3, wherein the transmission X-ray powder diffraction pattern further includes 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°.

[0099] Embodiment 6: The crystalline form according to Embodiment 3, wherein the transmission X-ray powder diffraction pattern further includes peaks at 12.0 ± 0.2° 2θ, 18.4 ± 0.2° 2θ, 19.8 ± 0.2° 2θ, and 21.7° 2θ ± 0.2°.

[0100] Embodiment 7: The crystalline form according to Embodiment 6, wherein the transmission X-ray powder diffraction pattern further includes 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°.

[0101] Embodiment 8: The crystalline form according to Embodiment 6, wherein the transmission X-ray powder diffraction pattern further includes peaks at 13.1 ± 0.2° 2θ, 14.4 ± 0.2° 2θ, 17.5 ± 0.2° 2θ, and 21.1° 2θ ± 0.2°.

[0102] Embodiment 9: The crystalline form according to any one of Embodiments 3 to 8, wherein the transmission X-ray powder diffraction is performed using Cu radiation.

[0103] Embodiment 10: The crystalline form according to any one of Embodiments 3 to 9, wherein the transmission X-ray powder diffraction is performed using a PANalytical Empyrean diffractometer operating in a transmission configuration, with a tube voltage of 45 kV and a filament emission of 40 mA.

[0104] Embodiment 11: The solid state includes 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 13The crystalline form according to any one of Embodiments 2 to 10, further characterized by a 13C NMR spectrum.

[0105] Embodiment 12: Solid state 13 The crystalline form according to Embodiment 11, wherein the 13C NMR spectrum includes peaks at 49.9 ± 0.2 ppm and 46.6 ± 0.2 ppm.

[0106] Embodiment 13: Solid state 13 The crystalline form according to Embodiment 11, wherein the 13C NMR spectrum includes 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.

[0107] Embodiment 14: Solid state 13 The crystalline form according to Embodiment 11, wherein the 13C NMR spectrum includes 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.

[0108] Embodiment 15: Solid state 13 The crystalline form according to Embodiment 11, wherein the 13C NMR spectrum includes 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.

[0109] Embodiment 16: Solid state 13The crystalline form according to Embodiment 11, wherein the 13C NMR spectrum includes 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.

[0110] Embodiment 17: Solid state 13 The crystalline form according to any one of Embodiments 11 to 16, wherein the 13C 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.

[0111] Embodiment 18: Solid state 13 The crystalline form according to any one of Embodiments 11 to 16, wherein the 13C NMR spectrum does not include peaks at 166.1 ± 0.2 ppm, 117.9 ± 0.2 ppm, and 108.6 ± 0.2 ppm.

[0112] Embodiment 19: Solid state 13 The crystalline form according to any one of Embodiments 11 to 16, wherein the 13C NMR spectrum does not include 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.

[0113] Embodiment 20: The crystalline form according to any one of Embodiments 2 to 19, further characterized by a differential scanning calorimetry curve including an endothermic melting peak starting at about 185 °C to about 200 °C.

[0114] Embodiment 21: The crystalline form according to Embodiment 20, wherein the endotherm starts at about 185 °C to about 197 °C.

[0115] Embodiment 22: The crystalline form of Embodiment 20, wherein endotherm starts at 191°C ± 5°C.

[0116] Embodiment 23: The crystalline form according to Embodiment 20, wherein the differential scanning calorimetry curve is substantially the same as the differential scanning calorimetry curve of FIG. 12.

[0117] Embodiment 24: The crystalline form according to any one of Embodiments 20 to 23, wherein the differential scanning calorimetry is performed using a TA Instruments Differential Scanning Calorimeter, Model Q2000, with the sample placed in an aluminum pan and heated to a temperature of 230°C at a rate of 10°C / min under nitrogen.

[0118] Embodiment 25: The crystalline form according to any one of Embodiments 3 to 24, further characterized by a thermogravimetric analysis thermogram showing a weight loss of less than about 0.5 wt% at about 25°C to about 110°C.

[0119] Embodiment 26: The crystalline form according to Embodiment 25, wherein the weight loss is less than about 0.2 wt%.

[0120] Embodiment 27: The crystalline form according to Embodiment 25, wherein the weight loss is less than about 0.1 wt%.

[0121] Embodiment 28: The crystalline form according to Embodiment 25, wherein the thermogravimetric analysis thermogram is substantially the same as the thermogravimetric analysis thermogram of FIG. 13.

[0122] Embodiment 29: The crystalline form according to any one of Embodiments 3 to 28, further characterized by a weight vapor sorption plot showing a reversible moisture uptake of less than about 0.5 wt% at about 25°C ± 0.1°C and from about 0% relative humidity to about 80% relative humidity.

[0123] Embodiment 30: The crystalline form according to Embodiment 29, wherein the reversible moisture uptake is less than about 0.2 wt%.

[0124] Embodiment 31: The crystalline form according to Embodiment 29, wherein the reversible water uptake is less than about 0.1% by weight.

[0125] Embodiment 32: The crystalline form according to Embodiment 29, wherein the weight vapor sorption plot is substantially the same as the weight vapor sorption plot of FIG. 14.

[0126] Embodiment 33: The crystalline form is as follows: a 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 13 a differential scanning calorimetry curve comprising an endothermic melting starting at about 185°C to about 200°C, further characterizing the crystalline form according to any one of Embodiments 3 to 10.

[0127] Embodiment 34: The crystalline form is as follows: a 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; 13 a differential scanning calorimetry curve comprising an endothermic melting starting at about 185°C to about 200°C; and a thermogravimetric analysis thermogram further characterizing the crystalline form according to any one of Embodiments 3 to 10, wherein the crystalline form exhibits a weight loss of less than about 0.2% by weight at about 25°C to about 110°C.

[0128] Embodiment 35: The crystalline form is as follows: a 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; 13 a differential scanning calorimetry curve comprising an endothermic melting starting at about 185°C to about 200°C; and The crystalline form according to any one of Embodiments 3 to 10, further characterized by a weight vapor sorption plot showing reversible water uptake of less than about 0.2 wt% at about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C.

[0129] Embodiment 36: The crystalline form is as follows: A solid state including 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 13 C NMR spectrum; A differential scanning calorimetry curve including an endothermic melting starting at 191°C ± 5°C; A thermogravimetric thermogram in which the crystalline form shows a weight loss of less than about 0.1 wt% at about 25°C to about 110°C; and The crystalline form according to any one of Embodiments 3 to 10, further characterized by a weight vapor sorption plot showing reversible water uptake of less than about 0.1 wt% at about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C.

[0130] Embodiment 37: The crystalline form according to any one of Embodiments 3 to 36, which is a crystalline anhydride.

[0131] Embodiment 38: The crystalline form according to any one of Embodiments 3 to 37, which contains less than 5 wt% of any other crystalline form of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide.

[0132] Embodiment 39: The crystalline form according to any one of Embodiments 3 to 37, which substantially does not contain any other crystalline form of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide.

[0133] Embodiment 40: The X-ray powder diffraction pattern includes 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°. The crystalline form according to Embodiment 2, wherein the X-ray powder diffraction pattern does not include peaks with relative intensities of medium intensity or higher at 18.7 ± 0.2° 2θ and 22.4° 2θ ± 0.2°.

[0134] Embodiment 41: The crystalline form according to Embodiment 40, wherein the X-ray powder diffraction pattern does not include peaks at 18.7 ± 0.2° 2θ and 22.4° 2θ ± 0.2°.

[0135] Embodiment 42: The crystalline form according to Embodiment 40, wherein the X-ray powder diffraction pattern further includes 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°.

[0136] Embodiment 43: The crystalline form according to Embodiment 40, wherein the X-ray powder diffraction pattern further includes peaks at 12.0 ± 0.2° 2θ, 18.4 ± 0.2° 2θ, 19.8 ± 0.2° 2θ, and 21.7° 2θ ± 0.2°.

[0137] Embodiment 44: The crystalline form according to Embodiment 43, wherein the X-ray powder diffraction pattern further includes 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°.

[0138] Embodiment 45: The crystalline form according to Embodiment 43, wherein the X-ray powder diffraction pattern further includes peaks at 13.1 ± 0.2° 2θ, 14.4 ± 0.2° 2θ, 17.5 ± 0.2° 2θ, and 21.1° 2θ ± 0.2°.

[0139] Embodiment 46: The crystalline form according to any one of Embodiments 40 to 45, wherein the X-ray powder diffraction is performed using Cu radiation.

[0140] Embodiment 47: The crystal form according to any one of Embodiments 40 to 46, wherein the transmission X-ray powder diffraction is performed using a PANalytical Empyrean diffractometer operating in a transmission configuration, a tube voltage of 45 kV, and a filament emission of 40 mA.

[0141] Embodiment 48: A solid state 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 13 The crystal form according to Embodiment 2 or any one of Embodiments 40 to 47, further characterized by a solid state 13C NMR spectrum.

[0142] Embodiment 49: A solid state 13 The crystal form according to Embodiment 48, wherein the solid state 13C NMR spectrum includes peaks at 166.1 ± 0.2 ppm, 117.9 ± 0.2 ppm, and 108.6 ± 0.2 ppm.

[0143] Embodiment 50: A solid state 13 The crystal form according to Embodiment 48, wherein the solid state 13C NMR spectrum includes 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.

[0144] Embodiment 51: A solid state 13 The crystal form according to Embodiment 48, wherein the solid state 13C NMR spectrum includes 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.

[0145] Embodiment 52: A solid state 13The crystalline form of any one of Embodiments 48 to 51, wherein the 13C 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.

[0146] Embodiment 53: Solid state 13 The crystalline form of any one of Embodiments 48 to 51, wherein the 13C 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.

[0147] Embodiment 54: Solid state 13 The crystalline form according to any one of Embodiments 48 to 51, wherein the 13C NMR spectrum does not include peaks at 49.9 ± 0.2 ppm and 46.6 ± 0.2 ppm.

[0148] Embodiment 55: Solid state 13 The crystalline form according to any one of Embodiments 48 to 51, wherein the 13C 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.

[0149] Embodiment 56: Solid state 13 The crystalline form according to any one of Embodiments 48 to 51, wherein the 13C 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.

[0150] Embodiment 57: A crystal form according to any one of Embodiments 2 or 40 - 56, further characterized by a differential scanning calorimetry curve including an endothermic melting having a start temperature of about 165°C to about 180°C.

[0151] Embodiment 58: The crystal form according to Embodiment 57, wherein the endotherm includes an endothermic melting having a start temperature of about 165°C to about 177°C.

[0152] Embodiment 59: The crystal form according to Embodiment 57, wherein the endotherm starts at 171°C ± 5°C.

[0153] Embodiment 60: The crystal form according to Embodiment 57, wherein the differential scanning calorimetry curve is substantially the same as the differential scanning calorimetry curve of Figure 6.

[0154] Embodiment 61: The crystal form according to any one of Embodiments 57 - 60, wherein the differential scanning calorimetry is performed with a TA Instruments Differential Scanning Calorimeter, model Q2000, with the sample placed in an aluminum pan and heated to a temperature of 230°C at a rate of 10°C / min under nitrogen.

[0155] Embodiment 62: A crystal form according to any one of Embodiments 40 - 61, further characterized by a thermogravimetric analysis thermogram in which the crystal form exhibits a weight loss of less than about 0.5 wt% at about 25°C to about 110°C.

[0156] Embodiment 63: The crystal form according to Embodiment 62, wherein the weight loss is less than about 0.2 wt%.

[0157] Embodiment 64: The crystal form according to Embodiment 62, wherein the weight loss is less than about 0.1 wt%.

[0158] Embodiment 65: The crystal form according to Embodiment 62, wherein the thermogravimetric analysis thermogram is substantially the same as the thermogravimetric analysis thermogram of Figure 7.

[0159] Embodiment 66: The crystalline form according to any one of Embodiments 40 to 65, further characterized by a weight vapor sorption plot showing a reversible water uptake of less than about 0.5 wt% at about 0% relative humidity to about 80% relative humidity at 25 °C ± 0.1 °C.

[0160] Embodiment 67: The crystalline form according to Embodiment 66, wherein the reversible water uptake is less than about 0.2 wt%.

[0161] Embodiment 68: The crystalline form according to Embodiment 66, wherein the reversible water uptake is less than about 0.1 wt%.

[0162] Embodiment 69: The crystalline form according to Embodiment 66, wherein the weight vapor sorption plot is substantially the same as the weight vapor sorption plot of FIG. 8.

[0163] Embodiment 70: The crystalline form is as follows: Solid state 13 C NMR spectrum including 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 13 C NMR spectrum; and The crystalline form according to any one of Embodiments 40 to 47, further characterized by a differential scanning calorimetry curve including an endothermic melting having an onset temperature of about 165 °C to about 180 °C.

[0164] Embodiment 71: The crystalline form is as follows: Solid state 13 C NMR spectrum including 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; 13 C NMR spectrum; Differential scanning calorimetry curve including an endothermic melting having an onset temperature of about 165 °C to about 177 °C; and The crystalline form according to any one of Embodiments 40 to 47, further characterized by a thermogravimetric analysis thermogram showing a weight loss of less than about 0.2 wt% at about 25 °C to about 110 °C.

[0165] Embodiment 72: The crystalline form is as follows: A solid state including 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 13 13C NMR spectrum; A differential scanning calorimetry curve including an endothermic melting peak having an onset temperature of about 165 °C to about 177 °C; and The crystalline form is further characterized by a gravimetric vapor sorption plot showing a reversible water uptake of less than about 0.2 wt% at about 0% relative humidity to about 80% relative humidity at 25 °C ± 0.1 °C, the crystalline form according to any one of Embodiments 40 to 47.

[0166] Embodiment 73: The crystalline form is as follows: A solid state including 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 13 13C NMR spectrum; A differential scanning calorimetry curve including an endothermic melting peak starting at 171 °C ± 5 °C; A thermogravimetric thermogram showing a weight loss of less than about 0.2 wt% at about 25 °C to about 110 °C for the crystalline form; and The crystalline form is further characterized by a gravimetric vapor sorption plot showing a reversible water uptake of less than about 0.2 wt% at about 0% relative humidity to about 80% relative humidity at 25 °C ± 0.1 °C, the crystalline form according to any one of Embodiments 40 to 47.

[0167] Embodiment 74: The crystalline form is a crystalline anhydride, the crystalline form according to any one of Embodiments 40 to 73.

[0168] Embodiment 75: The crystalline form contains less than 5 wt% of any other crystalline form of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinokynolin-4-carboxamide, the crystalline form according to any one of Embodiments 40 to 74.

[0169] Embodiment 76: The crystal form according to any one of Embodiments 40 to 74, wherein the crystal form substantially does not contain any other crystal form of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide.

[0170] Embodiment 77: The crystal form according to Embodiment 1, which is a crystal form of (R,S)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide.

[0171] Embodiment 78: The crystal form according to Embodiment 77, which is characterized by a reflection X-ray powder diffraction pattern including 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°.

[0172] Embodiment 79: The crystal form according to Embodiment 78, wherein the reflection X-ray powder diffraction pattern includes 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°.

[0173] Embodiment 80: The crystal form according to Embodiment 79, wherein the reflection X-ray powder diffraction pattern further includes 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°.

[0174] Embodiment 81: The crystal form according to Embodiment 79, wherein the reflection X-ray powder diffraction pattern further includes 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°.

[0175] Embodiment 82: The crystal form according to any one of Embodiments 77 to 81, wherein the reflection X-ray powder diffraction is performed using Cu radiation.

[0176] Embodiment 83: The crystal form according to any one of Embodiments 77 to 82, wherein the X-ray powder diffraction is performed using a PANalytical Empyrean diffractometer operating in a reflection configuration, with a tube voltage of 45 kV and a filament emission of 40 mA.

[0177] Embodiment 84: The crystal form according to any one of Embodiments 77 to 83, further characterized by a differential scanning calorimetry curve including an endothermic melting peak from about 195 °C to about 210 °C.

[0178] Embodiment 85: The crystal form according to Embodiment 84, wherein the endotherm has a starting temperature from about 195 °C to about 207 °C.

[0179] Embodiment 86: The crystal form according to Embodiment 84, wherein the endotherm has a starting temperature of 201 °C ± 5 °C.

[0180] Embodiment 87: The crystal form according to Embodiment 85, wherein the differential scanning calorimetry curve is substantially the same as the differential scanning calorimetry curve of Figure 16.

[0181] Embodiment 88: The crystal form according to any one of Embodiments 84 to 87, wherein the differential scanning calorimetry is performed using a TA Instruments Differential Scanning Calorimeter, model Q2000, with the sample placed in an aluminum pan and heated to a temperature of 230 °C at a rate of 10 °C / min under nitrogen.

[0182] Embodiment 89: The crystal form according to any one of Embodiments 77 to 88, further characterized by a gravimetric vapor sorption plot showing a reversible moisture uptake of less than about 1.0 wt% at 25 °C ± 0.1 °C from about 0% relative humidity to about 80% relative humidity.

[0183] Embodiment 90: The crystal form according to Embodiment 89, wherein the reversible moisture uptake is less than about 0.7 wt%.

[0184] Embodiment 91: The crystalline form according to Embodiment 89, wherein the reversible water uptake is about 0.5% by weight.

[0185] Embodiment 92: The crystalline form according to Embodiment 89, wherein the weight vapor sorption plot is substantially the same as the weight vapor sorption plot of FIG. 17.

[0186] Embodiment 93: The crystalline form is as follows: A differential scanning calorimetry curve including an endothermic melting having an onset temperature of about 190°C to about 210°C; and The crystalline form is further characterized by a weight vapor sorption plot showing a reversible water uptake of less than about 1.0% by weight at about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C, the crystalline form according to any one of Embodiments 78 to 92.

[0187] Embodiment 94: The crystalline form is as follows: A differential scanning calorimetry curve including an endothermic melting having an onset temperature of about 195°C to about 207°C; and The crystalline form is further characterized by a weight vapor sorption plot showing a reversible water uptake of less than about 0.7% by weight at about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C, the crystalline form according to any one of Embodiments 78 to 92.

[0188] Embodiment 95: The crystalline form is as follows: A differential scanning calorimetry curve including an endothermic melting starting at 201°C ± 5°C; and The crystalline form is further characterized by a weight vapor sorption plot showing a reversible water uptake of about 0.5% by weight at about 0% relative humidity to about 80% relative humidity at 25°C ± 0.1°C, the crystalline form according to any one of Embodiments 78 to 92.

[0189] Embodiment 96: The crystalline form according to any one of Embodiments 78 to 95, wherein the crystalline form is a crystalline anhydride.

[0190] Embodiment 97: The crystal form according to any one of Embodiments 78 to 96, wherein the crystal form contains less than 5% by weight of any other crystal form of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide.

[0191] Embodiment 98: The crystal form according to any one of Embodiments 78 to 96, wherein the crystal form substantially does not contain any other crystal form of N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide.

[0192] Embodiment 99: The crystal form according to any one of Embodiments 3 to 37; The crystal form according to any one of Embodiments 40 to 74; and A composition comprising at least two crystal forms selected from the group consisting of the crystal forms according to any one of Embodiments 77 to 96.

[0193] Embodiment 100: The crystal form according to any one of Embodiments 3 to 37; and The composition according to Embodiment 99, comprising the crystal form according to any one of Embodiments 40 to 74.

[0194] Embodiment 101: The composition according to Embodiment 100, comprising at least 50% by weight of the crystal form according to any one of Embodiments 3 to 37 with respect to the crystal form according to any one of Embodiments 40 to 74.

[0195] Embodiment 102: The composition according to Embodiment 100, comprising at least 75% by weight of the crystal form according to any one of Embodiments 3 to 37 with respect to the crystal form according to any one of Embodiments 40 to 74.

[0196] Embodiment 103: The composition according to Embodiment 100, comprising at least 90% by weight of the crystal form according to any one of Embodiments 3 to 37 with respect to the crystal form according to any one of Embodiments 40 to 74.

[0197] Embodiment 104: A composition according to Embodiment 100, comprising at least 95% by weight of a crystal form according to any one of Embodiments 3 to 37 with respect to the crystal form according to any one of Embodiments 40 to 74.

[0198] Embodiment 105: A composition according to Embodiment 100, comprising at least 95% by weight of a crystal form according to any one of Embodiments 3 to 37 with respect to any other crystal form.

[0199] Embodiment 106: A pharmaceutical composition comprising a crystal form according to any one of Embodiments 1 to 98 and one or more pharmaceutically acceptable excipients.

[0200] Embodiment 107: A pharmaceutical composition according to Embodiment 106, comprising a crystal form according to any one of Embodiments 3 to 37.

[0201] Embodiment 108: A pharmaceutical composition according to Embodiment 107, further comprising a crystal form according to any one of Embodiments 40 to 74.

[0202] Embodiment 109: A pharmaceutical composition according to Embodiment 106, comprising a crystal form according to any one of Embodiments 40 to 70.

[0203] Embodiment 110: A pharmaceutical composition according to any one of Embodiments 106 to 109, which is a solid pharmaceutical composition.

[0204] Embodiment 111: A method for treating or preventing a FAP-mediated condition in a subject suffering from or susceptible to a FAP-mediated condition, the method comprising administering to the subject a therapeutically effective amount of a crystal form according to any one of Embodiments 1 to 98.

[0205] Embodiment 112: The method according to Embodiment 111, wherein the FAP-mediated condition is selected from the group consisting of liver disease, type 2 diabetes mellitus, cardiovascular conditions, obesity, obesity-related conditions, fibrosis, keloid disorders, inflammation, and cancer.

[0206] Embodiment 113: The method according to embodiment 112, wherein the FAP-mediated condition is a liver disease.

[0207] Embodiment 114: The method according to embodiment 113, wherein the liver disease is non-alcoholic steatohepatitis.

[0208] 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 an FAP-mediated condition.

[0209] III. Method of Use (R)-N(2-(4-Cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-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.

[0210] FAP is thought to cleave and inactivate human fibroblast growth factor 21 (FGF-21) (Biochem. J. 2016, 473, 605), a protein involved in the regulation of glucose and lipid metabolism. Inhibition of FAP is hypothesized to increase endogenous FGF-21 levels and signaling, resulting in, for example, a reduction in adiposity, an improvement in insulin sensitivity, an improvement in glucose tolerance, a reduction in body weight, and / or a reduction in cardiovascular disease mortality.

[0211] FAP is also thought to cleave human α2-antiplasmin (α2AP), a protein involved in the regulation of fibrosis and fibrinolysis (Blood 2004 103,3783). Tissue repair involves coagulation leading to fibrin deposition. Fibrin in blood clots is usually lysed mainly by plasmin when converted from its inactive form (plasminogen) by plasminogen activators. 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 more effectively to fibrin, which reduces the plasminolysis of fibrin at the site of injury. Inhibition of FAP is hypothesized to increase fibrinolysis and improve tissue regeneration at the site of injury (J.Thromb.Haemost.2013,11,2029;Proteomics Clin.Appl.2014,8,454).

[0212] FAP is further thought to play a role in the increase of fibrosis by promoting collagen production and deposition and through changes in extracellular matrix (ECM) turnover (J Biol Chem 2016,8,291). Inhibition of FAP is hypothesized to result in a decrease in collagen deposition and a reduction in inflammation (Inflamm.Bowel Dis.2018,18,332).

[0213] Taking the above into account, inhibition of FAP is hypothesized to collectively reduce fibrosis and inflammation by decreasing hepatic stellate cell activity and increasing fibrinolysis, and further provide a positive metabolic effect through increased FGF21 signaling and improved glucose tolerance.

[0214] Accordingly, in some embodiments, the present disclosure provides a method for treating or preventing FAP-mediated conditions in a subject in need thereof, by administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of the crystalline form of the present disclosure.

[0215] In some embodiments, the present disclosure provides a method for treating or preventing a condition characterized by overexpression of FAP in a subject in need thereof, the method comprising administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure.

[0216] In some embodiments, the present disclosure provides a method for treating or preventing liver disease in a subject in need thereof, the method comprising administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure. 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, the NAFLD is selected from the group consisting of isolated steatosis, non-alcoholic steatohepatitis (NASH), hepatic fibrosis, and cirrhosis. In another aspect, the liver disease is end-stage liver disease. In another aspect, the subject also suffers from or is predisposed to one or more conditions selected from the group consisting of obesity, dyslipidemia, insulin resistance, type 2 diabetes, and renal insufficiency.

[0217] In some embodiments, the present disclosure provides a method for treating liver disease in a subject in need thereof, the method comprising 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 body mass index (BMI) of 2 ~40 kg / m 2 In one aspect, the subject has a BMI of 2 ~39.9 kg / m 2 In another aspect, the subject has a BMI of at least 2 40 kg / m. 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 hepatic fibrosis. In another aspect, the liver disease is cirrhosis.

[0218] 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 also has or is at risk of having 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.

[0219] 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 also has or is at risk of having 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.

[0220] 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 also has or is at risk of having 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.

[0221] 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 also has or is at risk of having 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.

[0222] In some embodiments, the present disclosure provides a method for treating a 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 is also suffering from or is susceptible to kidney failure. 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.

[0223] In some embodiments, the present disclosure provides a method for reducing liver fat 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 subject is suffering from or is susceptible to NAFLD. In another aspect, the subject is suffering from or is susceptible to NASH. In another aspect, the subject is suffering from or is susceptible to liver fibrosis. In another aspect, the subject is suffering from or is susceptible to cirrhosis. In another aspect, the subject is also suffering from or is susceptible to one or more conditions selected from the group consisting of obesity, dyslipidemia, insulin resistance, type 2 diabetes, and kidney failure.

[0224] In some embodiments, the present disclosure provides a method for treating or preventing non-alcoholic fatty liver disease (NAFLD) 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 NAFLD is stage 1 NAFLD. In another aspect, the NAFLD is stage 2 NAFLD. In another aspect, the NAFLD is stage 3 NAFLD. In another aspect, the 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 is also suffering 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.

[0225] In some embodiments, the present disclosure provides a method for treating or preventing non-alcoholic steatohepatitis (NASH) 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 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 is also suffering 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.

[0226] In some embodiments, the present disclosure provides a method for treating or preventing liver fibrosis 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 subject has stage 3 liver fibrosis. In another aspect, the subject also has or is susceptible to one or more conditions selected from the group consisting of obesity, dyslipidemia, insulin resistance, type 2 diabetes, and renal insufficiency.

[0227] In some embodiments, the present disclosure provides a method for treating or preventing cirrhosis 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 subject has stage F4 cirrhosis. In another aspect, the subject also has or is susceptible to one or more conditions selected from the group consisting of obesity, dyslipidemia, insulin resistance, type 2 diabetes, and renal insufficiency.

[0228] In some embodiments, the present disclosure provides a method for treating or preventing type 2 true diabetes 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 subject has diabetic kidney disease. In another aspect, the subject has renal insufficiency. In another aspect, the administration of the compound is adjunctive to diet and exercise. In another aspect, the administration of the compound also reduces body weight and / or treats obesity. In another aspect, the subject has a BMI of 27 kg / m 2 ~40 kg / m 2 In another aspect, the subject has a BMI of 30 kg / m 2 ~39.9 kg / 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.

[0229] In some embodiments, the present disclosure provides a method for improving glycemic control 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 subject has type 2 diabetes. In another aspect, the subject has diabetic kidney disease. In another aspect, the subject has renal insufficiency. In another aspect, the administration of the compound is adjunctive to diet and exercise. In another aspect, the administration of the compound also reduces body weight and / or treats obesity. In another aspect, the subject has a BMI of 27 kg / m 2 to 40 kg / m 2 . In another aspect, the subject has a BMI of 30 kg / m 2 to 39.9 kg / 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.

[0230] In some embodiments, the present disclosure provides a method for improving glycemic control in a subject having type 2 diabetes and diabetic kidney disease, 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 administration of the compound is adjunctive to diet and exercise. In another aspect, the administration of the compound also reduces body weight and / or treats obesity. In another aspect, the subject has a BMI of 27 kg / m 2 to 40 kg / m 2 . In another aspect, the subject has a BMI of 30 kg / m 2 to 39.9 kg / 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.

[0231] In some embodiments, the present disclosure provides a method for improving glycemic control in a subject having type 2 diabetes and renal insufficiency, 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 administration of the compound is an adjunct to diet and exercise. In another aspect, the administration of the compound also reduces body weight and / or treats obesity. In another aspect, the subject has a BMI of 27 kg / m 2 to 40 kg / m 2 . In another aspect, the subject has a BMI of 30 kg / m 2 to 39.9 kg / 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.

[0232] In some embodiments, the present disclosure provides a method for treating or preventing insulin resistance 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 another aspect, the subject has type 2 diabetes. In another aspect, the subject has diabetic kidney disease. In another aspect, the subject has renal insufficiency. Insulin resistance can be measured, for example, using the homeostasis model assessment of insulin resistance (HOMA-IR) and / or the Matsuda index. HOMA-IR is described, for example, in Diabetologia 1985, 28, 412, which is incorporated herein by reference in its entirety. The Matsuda index is described, for example, in Diabetes Care 1999, 22, 1462, which is incorporated herein by reference in its entirety.

[0233] In some embodiments, the present disclosure provides a method for treating or preventing impaired glucose tolerance 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 subject has type 2 diabetes. In another aspect, the subject has diabetic kidney disease. In another aspect, the subject has renal insufficiency.

[0234] In some embodiments, the present disclosure provides a method of treating a cardiovascular condition 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 cardiovascular condition is selected from the group consisting of heart failure, cardiomyopathy, atherosclerosis, venous thromboembolism, and atrial fibrillation. In one aspect, the cardiovascular condition is heart failure. In another aspect, the cardiovascular condition is heart failure with preserved ejection fraction (HFpEF). In another aspect, the cardiovascular condition 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 condition is atherosclerosis. In another aspect, the cardiovascular condition is venous thromboembolism. In another aspect, the cardiovascular condition is atrial fibrillation.

[0235] In some embodiments, the present disclosure provides a method of treating an obesity-related condition 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 obesity-related condition is an obesity-related metabolic condition. In another aspect, the obesity-related condition is selected from the group consisting of insulin resistance, prediabetes, type 2 diabetes, impaired glucose tolerance, elevated fasting blood glucose, and glucagonoma. In another aspect, the obesity-related condition is dyslipidemia. In another aspect, the obesity-related condition is a cardiovascular condition selected from the group consisting of heart failure, cardiomyopathy, atherosclerosis, venous thromboembolism, and atrial fibrillation. In another aspect, the obesity-related condition is a renal disease.

[0236] In some embodiments, the present disclosure provides a method of reducing body weight 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 subject has type 2 diabetes. In another aspect, the subject has diabetic kidney disease. In another aspect, the subject has renal insufficiency. In another aspect, the administration of the compound is adjunctive to diet and exercise. In another aspect, the administration of the compound also reduces body weight and / or treats obesity. In another aspect, the subject has a BMI of 27 kg / m 2 to 40 kg / m 2 . In another aspect, the subject has a BMI of 30 kg / m 2 to 39.9 kg / 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's body weight is reduced by, for example, at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, or 40%.

[0237] In some embodiments, the present disclosure provides a method of reducing body fat in a subject in need of treatment, 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 has type 2 diabetes. In another aspect, the subject has diabetic kidney disease. In another aspect, the subject has renal insufficiency. In another aspect, the administration of the compound is adjunctive to diet and exercise. In another aspect, the administration of the compound also reduces body weight and / or treats obesity. In another aspect, the subject has a BMI of 27 kg / m 2 to 40 kg / m 2 . In another aspect, the subject has a BMI of 30 kg / m 2 to 39.9 kg / 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 fat is liver fat.

[0238] In some embodiments, the present disclosure provides a method for treating or preventing fibrosis in a subject in need thereof, the method comprising administering to the subject 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).

[0239] In some embodiments, the present disclosure provides a method for promoting tissue remodeling in a subject in need thereof, the method comprising administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure. In one aspect, the subject has suffered cardiac tissue damage due to myocardial infarction.

[0240] In some embodiments, the present disclosure provides a method for promoting wound healing and / or reducing adhesions in a subject in need thereof, the method comprising 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.

[0241] In some embodiments, the present disclosure provides a method for treating or preventing keloid disorders in a subject in need thereof, the method comprising 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 scar formation, keloid tumors, and keloid scars.

[0242] In some embodiments, the present disclosure provides a method for treating or preventing inflammation in a subject in need thereof, the method comprising administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of a crystalline form of the present disclosure. In one aspect, the inflammation is 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.

[0243] In some embodiments, the present disclosure provides a method of treating cancer 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 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, pharyngeal cancer, multiple myeloma cells, bladder cancer, cholangiocarcinoma, renal clear cell carcinoma, neuroendocrine tumor, tumor-induced osteomalacia, sarcoma, CUP (cancer of unknown primary), thymic carcinoma, desmoid tumor, myxoma, astrocytoma, cervical cancer, and prostate cancer. In another aspect, the cancer is hepatocellular carcinoma.

[0244] The subject to be treated is typically a human or non-human mammal, particularly a human. Suitable subjects may include domestic or wild animals; companion animals (including dogs, cats, etc.); livestock (including horses, cows and other ruminants, pigs, poultry, rabbits, etc.); primates (including monkeys such as rhesus monkeys, cynomolgus monkeys (also known as cynomolgus monkeys or long-tailed macaques), marmosets, tamarins, chimpanzees, macaques, etc.); and rodents (including rats, mice, gerbils, guinea pigs, etc.).

[0245] In some embodiments, the present disclosure provides a crystalline form of the present disclosure for use as a medicament.

[0246] In some embodiments, the present disclosure provides the use of a crystalline form of the present disclosure for treating or preventing the above-mentioned FAP-mediated conditions.

[0247] In some embodiments, the present disclosure provides the use of a crystalline form of the present disclosure for the manufacture of a medicament for treating or preventing the above-mentioned FAP-mediated conditions.

[0248] IV. Combination Therapy and Fixed Dose Combinations The crystalline forms of the present disclosure can be used in the above methods either as a single pharmacological agent or in combination with other pharmacological agents or techniques. Such combination therapies can be achieved by administering the individual components of the treatment simultaneously, sequentially, or separately. These combination therapies (and corresponding combination products) use the crystalline forms of the present disclosure within the dosage ranges described herein and other pharmacological agents, typically other pharmacological agents within their approved dosage ranges.

[0249] In some embodiments, the present disclosure provides a combination suitable for use in the treatment of a condition selected from the foregoing conditions, the combination comprising a crystalline form of the present disclosure and a sodium-glucose cotransporter 2 (SGLT2) inhibitor. In one aspect, the SGLT2 inhibitor is selected from the group consisting of canagliflozin, dapagliflozin, empagliflozin, ertugliflozin, ipragliflozin, luseogliflozin, and remogliflozin. In another aspect, the SGLT2 inhibitor is dapagliflozin.

[0250] In some embodiments, the present disclosure provides a combination suitable for use in the treatment of a condition selected from the foregoing conditions, the combination comprising a crystalline form of the present disclosure and metformin.

[0251] In some embodiments, the present disclosure provides a combination suitable for use in the treatment of a condition selected from the foregoing conditions, the combination comprising 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.

[0252] In some embodiments, the present disclosure provides a combination suitable for use in the treatment of a condition selected from the foregoing conditions, the combination comprising 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, gemigliptin, anagliptin, teneligliptin, alogliptin, treagliptin, omarigliptin, evogliptin, gosogliptin, and dutogliptin.

[0253] In some embodiments, the present disclosure provides a combination suitable for use in the treatment of a condition selected from the foregoing conditions, the combination comprising 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 riboglitazone. In another aspect, the PPAR agonist stimulates hepatic expression of FGF21.

[0254] In some embodiments, the disclosure provides a pharmaceutical composition comprising a crystalline form of the disclosure, one or more pharmacological agents selected from SGLT2 inhibitors, metformin, GLP1 agonists, DPP4 inhibitors, and PPAR agonists, and a pharmaceutically acceptable diluent or carrier. Such combinations can be used in the manufacture of a medicament for use in the treatment of a condition selected from the foregoing conditions. 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.

[0255] In some embodiments, the disclosure provides a combination suitable for use in the treatment of cancer, the combination comprising a crystalline form of the 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, Lag-3 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, the cancer is selected from the group consisting of pancreatic cancer, colon cancer, and rectal cancer.

[0256] V. Pharmaceutical Composition The crystalline forms of the disclosure can be administered as a pharmaceutical composition comprising one or more pharmaceutically acceptable excipients. Thus, in some embodiments, the disclosure provides a pharmaceutical composition comprising a crystalline form of the disclosure and at least one pharmaceutically acceptable excipient.

[0257] The excipients selected for inclusion in a particular composition will vary depending on factors such as the mode of administration and the form of the composition to be provided. Suitable pharmaceutically acceptable excipients are well known to those skilled in the art and are described, for example, in the Handbook of Pharmaceutical Excipients, Sixth Edition, Pharmaceutical Press, edited by Rowe, Ray C; Sheskey, Paul J; Quinn, Marian. Pharmaceutically acceptable excipients can function, for example, as adjuvants, diluents, carriers, stabilizers, flavoring agents, coloring agents, fillers, binders, disintegrants, lubricants, glidants, thickeners, and coating agents. As will be understood by those skilled in the art, a particular pharmaceutically acceptable excipient can perform two or more functions and can perform alternative functions depending on the amount of excipient present in the composition and the other excipients present in the composition.

[0258] The composition may be in a form suitable for oral use (for example, 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 (for example, as creams, ointments, gels, or aqueous or oily solutions or suspensions), in a form suitable for administration by inhalation (for example, as fine powders or liquid aerosols), in a form suitable for administration by insufflation (for example, as fine powders), or in a form suitable for parenteral administration (for example, as sterile aqueous or oily solutions for intravenous, subcutaneous or intramuscular administration), or in a form suitable as a suppository for rectal administration. The composition can be obtained by conventional procedures using conventional pharmaceutically excipients well known in the art. Thus, a composition intended for oral use may contain, for example, one or more coloring agents, sweetening agents, flavoring agents and / or preservatives.

[0259] The total daily dosage will necessarily vary depending on the subject being treated, the route of administration, any therapies co-administered, and the severity of the disease being treated, and may include a single dose or multiple doses. Specific dosages can be adjusted, for example, according to the condition being treated; the age, weight, general health, gender, and diet of the subject; the route of administration; the dosing interval; the rate of excretion; and other drugs co-administered to the subject. A normally skilled physician provided with the disclosure herein can determine the appropriate dosage and regimen for the administration of the therapeutic agent to the subject according to methods well known in the art of therapy and can adjust such dosage and regimen as necessary during the course of treatment. The crystalline forms of the present disclosure are typically administered to warm-blooded animals at unit dosages within the range of 2.5 to 5000 mg / m 2 of the body surface area of the animal, or within the range of about 0.05 to 100 mg / kg, which typically provides a therapeutically effective dosage.

[0260] 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.

[0261] In some embodiments, the present disclosure provides a pharmaceutical composition for use in the treatment of FAP-mediated conditions, comprising a crystalline form of the present disclosure and at least one pharmaceutically acceptable excipient. In one aspect, the FAP-mediated conditions are selected from the group consisting of liver disease, type 2 diabetes mellitus, cardiovascular conditions, obesity, obesity-related conditions, fibrosis, keloid disorders, inflammation, and cancer.

[0262] 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.

[0263] 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.

[0264] 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.

[0265] In some embodiments, the present disclosure provides a pharmaceutical composition comprising Form 2 and one or more pharmaceutically acceptable excipients. In one aspect, the composition comprises at least 90% by weight of Form 2 relative to any other crystalline form. In another aspect, the composition comprises at least 95% by weight of Form 2 relative to any other crystalline form. In another aspect, the composition comprises at least 96% by weight of Form 2 relative to any other crystalline form. In another aspect, the composition comprises at least 97% by weight of Form 2 relative to any other crystalline form. In another aspect, the composition comprises at least 98% by weight of Form 2 relative to any other crystalline form. In another aspect, the composition comprises at least 99% by weight of Form 2 relative to any other crystalline form.

[0266] VI. Kit The present disclosure further provides a kit comprising a unit dosage form containing a crystalline form of the present disclosure housed within a packaging material, and a label or package insert indicating that the unit dosage form can be used to treat one or more of the foregoing conditions.

[0267] In some embodiments, the kit comprises a unit dosage form containing a crystalline form of the present disclosure housed within a packaging material, and a label or package insert indicating that the pharmaceutical composition can be used to treat FAP-mediated conditions. In one aspect, the FAP-mediated condition 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).

[0268] In some embodiments, the kit comprises: (a) a first unit dosage form containing a crystalline form of the present disclosure; (b) a second unit dosage form containing a pharmacological agent selected from the group consisting of an SGLT2 inhibitor, metformin, a GLP1 agonist, a DPP4 inhibitor, and a PPAR agonist; (c) container means for housing the first and second dosage forms; 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 FAP-mediated conditions.

Example

[0269] VII. Example The following descriptions of experiments, procedures, examples, and intermediates are intended to illustrate embodiments of the present disclosure. These are by no means intended to be limiting. Other embodiments of the present disclosure can be prepared using either the methods exemplified in these examples alone or in combination with techniques generally known in the art.

[0270] Example 1: Preparation of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide

[0271]

Chemical formula

[0272] (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide can be prepared as described below.

[0273] A. (R)-3-glycylthiazolidine-4-carbonitrile hydrochloride (Intermediate 1)

[0274]

Chemical formula

[0275]

Chemical formula

[0276] Di-tert-butyl dicarbonate (Boc2O, 18.6 mL, 80.2 mmol) was added to a stirred solution of (R)-3-(tert-butoxycarbonyl)thiazolidine-4-carboxylic acid (17.0 g, 72.9 mmol) and pyridine (7.07 mL, 87.5 mmol) in ethyl acetate (170 mL), and the reaction mixture was stirred at room temperature for 3 h. Next, a solution of NH3 (25% aqueous solution, 6 mL) was added dropwise, and the mixture was stirred at room temperature overnight. The reaction mixture was diluted with ethyl acetate, the phases were separated, the organic phase was washed with saturated NaCl, dried, filtered through a pad of silica gel, washed with ethyl acetate, and evaporated to give crude tert-butyl (R-4-carbamoylthiazolidine-3-carboxylate (Intermediate 1-A, 16.9 g, 100%) as a colorless oil, which was used directly in the next step.

[0277] (ii) tert-butyl (R)-4-cyanothiazolidine-3-carboxylate (Intermediate 1-B)

[0278] [Chemical formula]

[0279] Trifluoroacetic anhydride (TFAA, 12.4 mL, 87.5 mmol) as a solution in ethyl acetate (20 mL) was added at room temperature to a solution of crude tert-butyl (R-4-carbamoyl-thiazolidine-3-carboxylate (Intermediate 1-A, 16.9 g, 72.9 mmol) and pyridine (14.7 mL, 182 mmol) in ethyl acetate (150 mL). The mixture was stirred at room temperature for 4 h, then diluted with ethyl acetate and washed with aqueous HCl (1 M), saturated NaHCO3. The organic phase was dried, filtered through a pad of silica gel, washed with ethyl acetate, and evaporated to give a pale yellow oil, which was allowed to stand and solidify. The crude solid material was suspended in heptane:ethyl acetate (4:1, 50 mL) and stirred at room temperature overnight. The solid was filtered off, washed with heptane:ethyl acetate (4:1), and dried to give tert-butyl (R)-4-cyanothiazolidine-3-carboxylate (Intermediate 1-B, 12.0 g, 83%) as a colorless solid; 11H NMR (400 MHz, CDCl3) δ 5.20 - 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).

[0280] (iii) (R)-Thiazolidine-4-carbonitrile hydrochloride (Intermediate 1-C)

[0281]

Chem.

[0282] HCl (12 M, 11 mL) in methanol (140 mL) was slowly added to a solution of tert-butyl (R)-4-cyanothiazolidine-3-carboxylate (Intermediate 1-B, 6.0 g, 28 mmol) in methanol (140 mL) at room temperature. The colorless transparent 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 1H NMR (400 MHz, CD3OD) δ 4.90 (dd, 1H), 4.35 - 4.24 (m, 2H), 3.37 - 3.24 (m, 2H).

[0283] (iv) tert-Butyl (R)-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)carbamate (Intermediate 1-D)

[0284]

Chem.

[0285] 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 propane phosphonic anhydride (T3P, 41.6 mL, 70.0 mmol, 50% solution in ethyl acetate) in ethyl acetate (120 mL). The mixture was heated at 60 °C for 4 h. The mixture was diluted with ethyl acetate and washed successively with water, HCl (1 M) aqueous solution 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 (400 MHz, CDCl3) δ 5.36 - 5.25 (m, 2H), 4.59 - 4.52 (m, 2H), 4.14 - 3.90 (m, 2H), 3.29 (d, 2H), 1.45 (s, 9H).

[0286] (v) (R)-3-Glycylthiazolidine-4-carbonitrile hydrochloride (Intermediate 1)

[0287]

Chem.

[0288] HCl (12 M, 5.6 mL) in methanol (140 mL) was slowly added to a solution of tert-butyl (R)-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)carbamate (Intermediate 1-D, 7.60 g, 28.0 mmol) in methanol (140 mL), and then 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; 11H NMR (400 MHz, CD3OD) δ 5.34 (t, 1H), 4.72 (d, 1H), 4.62 (d, 1H), 4.11 - 3.94 (m, 2H), 3.41 - 3.36 (m, 2H).

[0289] B. 6 - Morpholinquinoline - 4 - carboxylic acid (Intermediate 2)

[0290]

Chem.

[0291]

Chem.

[0292] 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 successively with water and brine. The organic layer was dried by passing through a phase separator and concentrated under reduced pressure to give ethyl 6 - morpholinquinoline - 4 - carboxylate (Intermediate 2 - A, 110 mg, 30%); MS m / z (ESI), [M + H]+ 287.2.

[0293] (ii) 6 - Morpholinquinoline - 4 - carboxylic acid (Intermediate 2)

[0294]

Chem.

[0295] NaOH (31 mg, 0.77 mmol) was added to a solution of ethyl 6-morpholinoquinoline-4-carboxylate (Intermediate 2-A, 110 mg, 0.38 mmol) in methanol (4 mL), and the mixture was heated at 60 °C for 2 h. 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.

[0296] C. (R)-N-(2-(4-Cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide

[0297]

Chemical formula

[0298] N-Ethyl-N-isopropyl-propan-2-amine (DIPEA, 0.15 mL, 0.87 mmol) was added to a suspension of 6-morpholinoquinoline-4-carboxylic acid (Intermediate 2, 75 mg, 0.29 mmol), (R)-3-glycylthiazolidine-4-carbonitrile hydrochloride (Intermediate 1, 121 mg, 0.58 mmol), 1-hydroxybenzotriazole hydrate (HoBt, 53 mg, 0.35 mmol), and 3-(ethyliminomethyleneamino)-N,N-dimethyl-propan-1-amine hydrochloride (EDC, 84 mg, 0.44 mmol) in ethyl acetate (1 mL) and acetonitrile (1 mL). A clear yellow solution was obtained, which was stirred at room temperature overnight. The mixture was diluted with ethyl acetate and washed with saturated NaHCO3 and brine. The organic phase was 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 H2O / acetonitrile / formic acid (95 / 5 / 0.2) (from 5% to 65%) as the mobile phase to give (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide (26 mg, 21%); C20 H 22 HRMS (ESI) m / z of N5O3S [M+H]+: calculated value 412.1438, measured value 412.1437; 1 1H NMR (400 MHz, CD3CN) δ 8.71 (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).

[0299] Example 2: Analysis Method Unless otherwise specified, the following analysis methods were used to characterize the crystal forms described in the following examples.

[0300] A. Reflective Powder X-ray Diffraction (Amorphous PXRD) X-ray diffraction analysis was performed according to standard methods, which can be found, for example, in Kitaigorodsky, A.I. (1973), Molecular Crystals and Molecules, Academic Press, New York; Bunn, C.W. (1948), Chemical Crystallography, Clarendon Press, London; or Klug, H.P. & Alexander, L.E. (1974), X-ray Diffraction Procedures, John Wiley & Sons, New York.

[0301] The powder X-ray diffraction (referred to as PXRD in this specification) pattern was determined by placing the sample on a zero background holder, single crystal silicon, and spreading the sample into a thin layer. PXRD was recorded using a Theta-Theta PANalytical X’Pert PRO (X-ray wavelength 1.5418 Å nickel filtered Cu radiation, voltage 45 kV, filament emission 40 mA). Variable divergence and anti-scatter slits and incident and diffracted soller slits of 0.04° were used. During the measurement, the sample was rotated. The sample was scanned from 2.4 to 50° 2θ using a PIXcel1D detector (effective length 3.35° 2θ), a step width of 0.013° and a count time of 115.770 seconds. The PXRD pattern was obtained in a Bragg-Brentano configuration.

[0302] One of ordinary skill in the art will recognize that a PXRD pattern with one or more measurement errors can be obtained depending on the measurement conditions such as the apparatus or machine used (Jenkins, R & Snyder, R.L. ‘Introduction to X-Ray Powder Diffractometry’ John Wiley & Sons 1996; Bunn, C.W. (1948), Chemical Crystallography, Clarendon Press, London; Klug, H.P. & Alexander, L.E. (1974), X-Ray Diffraction Procedures). One of ordinary skill in X-ray powder diffraction will further recognize that the relative intensity of the peaks can be affected by, for example, particles with a size greater than 30 microns and non-unitarial aspect ratios that can affect the analysis of the sample. Further, it should be understood that the intensity can vary depending on the experimental conditions and sample preparation (e.g., preferred orientation). The following definitions were used for relative intensity (%): 25% - 100%, vs (very strong); 10% - 25%, s (strong); 3% - 10%, m (medium strong); 1% - 3%, w (weak).

[0303] One skilled in the art will also recognize that the position of the reflection can be affected by the exact height at which the sample is positioned within the diffractometer and the zero calibration of the diffractometer. The surface flatness of the sample can also have a minor effect. Therefore, the presented diffraction pattern data should not be interpreted as absolute values. Generally, the measurement error of the diffraction angle in an X-ray powder diffractogram can be approximately ±0.2° 2θ, and the extent of such measurement error should be considered when examining PXRD data.

[0304] The reflection mode PXRD pattern can be compared to the transmission mode PXRD pattern, and one skilled in the art will understand that the diffraction pattern can vary, particularly with respect to peak intensity.

[0305] B. Reflection Powder X-ray Diffraction (Type 2 PXRD) The powder X-ray diffraction (referred to herein as PXRD) pattern was determined by placing the sample on a zero background holder on a single crystal silicon and spreading the sample into a thin layer. Powder X-ray diffraction was recorded using a reflection Theta-Theta PANalytical Empyrean (X-ray wavelength 1.5419 Å nickel filtered Cu radiation, voltage 45 kV, filament emission 40 mA). Variable divergence and anti-scattering slits and incident and diffracted soller slits of 0.04° were used. During the measurement, the sample was rotated. The sample was scanned from 2.4 to 50° 2θ using a PIXcel3D-Medipix3 detector (effective length 3.35° 2θ) with a step width of 0.013° and a count time of 234.345 seconds. The following definitions were used for relative intensity (%): 25% - 100%, vs (very strong); 10% - 25%, s (strong); 3% - 10%, m (medium strong); 1% - 3%, w (weak).

[0306] C. Transmission Powder X-ray Diffraction (PXRD of Forms A and B) Powder X-ray diffraction data are measured using corundum as an internal standard. The powder X-ray diffraction (PXRD) pattern is determined by mounting the sample between two Kapton® polyimide films to form a thin layer between the films. PXRD is recorded using a transmission PANalytical Empyrean (X-ray wavelength 1.5419 Å nickel-filtered Cu radiation, voltage 45 kV, filament emission 40 mA). Fixed divergence and anti-scatter slits are used, and the sample is rotated during measurement. The sample is scanned from 2.5 to 50° 2θ using a PIXcel3D-Medipix3 detector (effective length 3.35° θ) with a 0.013° step width and a 938.145 s count time. The following definitions were used for relative intensity (%): 25% - 100%, vs (very strong); 10% - 25%, s (strong); 3% - 10%, m (medium strong); 1% - 3%, w (weak).

[0307] D. Solid State 13 C NMR Spectroscopy Approximately 100 mg of the material is loaded into a 4 mm zirconium dioxide rotor sealed with a Kel-F cap. 13 For the determination of the C cross-polarization Magic Angle Spinning spectrum, the rotor is spun at 15 kHz (to remove chemical shift anisotropy), and cross-polarization from hydrogen is used 13 to record the C spectrum (to improve sensitivity and shorten the experimental time). The contact time for magnetization transfer is 2 milliseconds, and the inter-pulse delay (to allow nuclear relaxation) is 15 seconds. Signal averaging is used by recording sufficient scans (typically 1k scans) to enable all major peaks to be resolved from the noise. Glycine is used as a secondary chemical shift reference (main peak at 176.03 ppm).

[0308] E. Lamp Differential Scanning Calorimetry (DSC) Melting onset temperature (T m) is determined by differential scanning calorimetry using a TA Instruments DSC, model Q2000. A sample (about 1 - 3 mg) is weighed into an aluminum sample pan. The sample is packed at the bottom of the sample pan and a lid with a pinhole is used. The apparatus is purged with nitrogen at 50 mL / min and data is collected at 25 °C and from 210 - 250 °C using a heating rate of 10 °C / min.

[0309] F. MDSC Differential Scanning Calorimetry (DSC) The glass transition temperature midpoint (T g ) is determined by MDSC differential scanning calorimetry using a TA Instruments DSC, model Q2000. A sample (about 5 mg) is weighed into an aluminum sample pan. The sample is packed at the bottom of the sample pan and a lid with a pinhole is used. The apparatus is purged with nitrogen at 50 mL / min and data is collected from 25 °C to 230 °C using a base heating rate of 5 °C / min and a heat-only modulation method with a modulation amplitude of ±0.53 °C every 40 seconds.

[0310] G. Thermogravimetric Analysis (TGA) Thermogravimetric analysis is performed using a TA Instruments TGA, model Q500. A sample (about 10 mg) is transferred to a sample holder weighed in tare. The apparatus is purged with nitrogen, oven at 60 mL / min and balance at 40 mL / min and data is collected from room temperature to 300 °C using a heating rate of 10 °C / min. During heating, a weight gain is observed due to the buoyancy effect. This effect can be reduced by using a material over 15 mg or by performing baseline subtraction on the sample curve.

[0311] H. Gravimetric Vapor Sorption (GVS) Weight vapor sorption analysis is performed using a TA Instruments TGA, model Q5000SA. The sample (approximately 5 - 10 mg) is transferred to a pre-weighed sample holder. The apparatus is purged with nitrogen at 25 °C at 200 mL / min for the chamber and 10 mL / min for the balance, and data is collected at different relative humidities (%RH). Starting at 20%RH, it is increased stepwise to 80%RH, decreased stepwise to 0%RH, and finally increased to 90%RH and returned to 0%RH in a second cycle. The equilibrium criterion for moving to the next %RH is achieved when the drift criterion (dm / dt) is less than 0.002 for 10 minutes.

[0312] Hygroscopicity can be evaluated, 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: sufficient water is absorbed to form a liquid; all values were measured as weight gain at 80% RH and 25 °C).

[0313] Example 3: Mixture of Form A / Form B Initial crystallization attempts to isolate pure Form A and pure Form B often resulted in a mixture of Form A and Form B. Crystallization parameters such as temperature, time, and solvent composition affected the type and quality of the final crystalline product obtained.

[0314] A. 50 / 50 mixture In this test, when (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide was isolated from a mixture of isopropyl alcohol and ethyl acetate, a mixture of Form A and Form B (~50 / 50) was obtained. Specifically, 6-morpholino-quinoline-4-carboxylic acid (Intermediate 2 of Example 1, 177 g, 0.757 mol, 1.0 equivalent) was placed in a 5 L jacketed vessel. (R)-3-glycylthiazolidine-4-carboxamide (159 g, 1.2 equivalents), and then ethyl acetate (3500 mL) were added to the reactor. Subsequently, a 50% wt / wt solution of propylphosphonic anhydride in ethyl acetate (1310 g, 3 equivalents), and then DIPEA (443 g, 5.002 equivalents) were added to the reactor. The temperature was adjusted to 65 °C, and the reaction mixture was stirred for 48 hours and then cooled to 20 °C.

[0315] The reaction mixture was combined with a second reaction mixture (465 g in total) prepared in the same manner and placed in a 50 L vessel for work-up. Ethyl acetate (20 L) was added to the vessel, and the pH was adjusted to approximately 9 using 10 L of saturated aqueous NaHCO3. The resulting two-phase mixture was stirred at 20 °C for 15 minutes, and then the layers were separated. The aqueous phase was extracted with 10 L of ethyl acetate, stirred at 20 °C for 15 minutes, and then the phases were separated. The organic phase was washed with 10 L of saturated aqueous NaHCO3, stirred at 20 °C for 15 minutes, and then the layers were separated. All the organic phases were combined and concentrated to approximately 5 volumes at 45 °C using a rotary evaporator. The organic phase was passed through a 5 kg silica pad using isopropanol (15 L) as the eluent. Subsequently, the organic phase was concentrated to approximately 5 volumes at 45 °C using a rotary evaporator, the concentrated organic phase was replaced with 10 L of ethyl acetate, and concentrated to approximately 8 volumes. The suspension was filtered, and the wet cake was washed with 2 L of ethyl acetate. The wet cake was dried at 50 °C for 18 hours to obtain a pale yellow solid, which is an approximately 50 / 50 mixture of Form A and Form B of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide.

[0316] B. Additional Tests A small-scale crystallization test was carried out. The amorphous (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholino-quinoline-4-carboxamide was suspended or dissolved (20 relative volumes) in each of the solvents listed in Table 1 below and stirred at 20 °C for 3 days. Subsequently, each of the resulting solids was analyzed by PXRD and DSC. The results are reported in Table 1 below.

[0317]

Table 1

[0318] The use of ethyl acetate or methyl isobutyl ketone as the crystallization solvent appeared to be favorable for the formation of Form A, while the use of acetone, methyl ethyl ketone, or acetonitrile appeared to be favorable for the formation of Form B. However, the other crystallization parameters selected also appear to affect the resulting crystal form. In Example 3A above, a 50 / 50 mixture of Form A and Form B was produced by a solvent system containing isopropanol and ethyl acetate. However, the low temperature and limited time used during the concentration procedure may not have been 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 containing acetone and ethyl acetate. The suspension was stirred at an elevated temperature for several hours, which allowed most of the lower melting Form A to be dissolved, and then the suspension was concentrated over time with the higher melting Form B. When the suspension was cooled to 20 °C, the existing Form B acted as a seed crystal.

[0319] Example 4: Crystal Form A A. Preparation of Crystal Form A Amorphous (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholino-quinoline-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 and slowly cooled to 20 °C over 5 hours, and then stirred at 20 °C for an additional 12 hours. The fine suspension was filtered, and the yellow solid was washed with ethyl acetate (2 × 50 mL). The collected solid was aspirated first under vacuum at 20 °C for 10 minutes, then at 40 °C for 2 hours in a relatively dry state, and finally dried under vacuum at 35 °C over the weekend. PXRD and DSC analyses showed mainly pure Form A.

[0320] B. Physical Characterization of Crystal Form A The characterization of Form A was carried out using various techniques including PXRD (Figure 1), solid-state 13 13C NMR spectroscopy (Figures 2 - 5), differential scanning calorimetry (DSC) (Figure 6), thermogravimetric analysis (TGA) (Figure 7), and gravimetric vapor sorption (GVS) (Figure 8).

[0321] The PXRD pattern in Figure 1 confirms that Form A is crystalline. Figure 1 shows the PXRD pattern of Form A measured using a transmission configuration. In Table 2 below, the selected peaks identified in the PXRD pattern of Figure 1 are listed.

[0322]

Table 2

[0323] Form A shows characteristic peaks at 12.0 ± 0.2° 2θ, 18.4 ± 0.2° 2θ, 19.8 ± 0.2° 2θ, and 21.7° 2θ ± 0.2° 2θ (compared to other forms excluding Form B). Form A shows additional 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 characteristic peaks (i.e., relative intensity peaks with medium intensity or above) at 18.7 ± 0.2° 2θ and 22.4° 2θ ± 0.2° 2θ compared to Form B.

[0324] Figure 2 shows the representative solid state of a sample of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide that is approximately 95 wt% Form A and 5 wt% Form B. 13 13C NMR spectrum is shown. Figure 3 shows the representative solid state of a sample of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide that is approximately 30 wt% Form A and 70 wt% Form B. 13 13C NMR spectrum is shown. Figure 4 shows the representative solid state of a sample of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide that is Form B with a small amount of Form A. 13 13C NMR spectrum is shown. Figure 5 is a comparison of the solid state 13C NMR spectra of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide Forms A and B based on the analysis of the combined spectra of Figures 2, 3, and 4. 13 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 characteristic 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 that are characteristic compared to Form B.

[0325] Figure 6 shows a representative lamp DSC thermogram of Form A. The exothermic events are plotted upward. The endothermic melting shown in Figure 6 has an onset temperature of about 171 °C and a heat enthalpy of about 68 J / g for the endothermic melting. The DSC values obtained may vary by about ±5 °C depending on the apparatus used, the method by which the sample is prepared, and differences between batches.

[0326] Figure 7 shows a representative TGA thermogram of Form A. Form A showed a weight loss of less than about 0.1% upon heating from about 25 °C to 110 °C, confirming that Form A is an anhydrate.

[0327] Figure 8 shows a representative GVS plot of Form A. Form A showed a reversible moisture uptake of about 0.05 wt% at 25 °C ± 0.1 °C from 0% relative humidity to 80% relative humidity. The desorption curve indicates that Form A lost moisture at a similar rate as the moisture taken up during sorption, with limited hysteresis. No morphological changes by PXRD were observed after the GVS experiment. According to the European Pharmacopoeia (EP) classification, Form A is non-hygroscopic (i.e., <0.2% weight gain).

[0328] Example 5: Crystal Form B A. Preparation of Crystal Form B (i) Crystallization without seed (acetone) Amorphous (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholino-quinoline-4-carboxamide (30 mg) was dissolved in 0.4 mL of acetone. After 30 minutes, when the solution became gradually viscous, additional acetone (0.2 mL) was added to obtain a fine suspension. The suspension was stirred for 3 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.

[0329] (ii) Crystallization without seed (methyl ethyl ketone) Amorphous (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholino-quinoline-4-carboxamide (30 mg) was dissolved in 0.4 mL of methyl ethyl ketone. After 15 minutes, when the solution became gradually viscous, further methyl ethyl ketone (0.2 mL) was added to obtain a fine suspension. The suspension was stirred for 3 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.

[0330] (iii) Crystallization without seed crystal (acetonitrile) Amorphous (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholino-quinoline-4-carboxamide (30 mg) was dissolved in 0.4 mL of acetonitrile. After 45 minutes, when the solution became gradually viscous, and after 2 hours, further acetonitrile (0.2 mL) was added to obtain a fine suspension. The suspension was stirred for 3 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.

[0331] (iv) Crystallization with seed crystal addition (acetonitrile) (R)-N-(2-(4-Cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-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 filtrate was cooled to 40 °C at a rate of 1.5 °C / min or less. To the solution was added seed crystal of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide Form B (150 mg / 0.025 equivalent, pre-ground with a pestle and mortar), and then the mixture was 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 isolated by filtration under vacuum. The filter cake was washed successively with 6 mL of acetonitrile and 12 mL of tert-butyl methyl ether, and then dried at 40 °C under vacuum to obtain the final product of crystalline Form B (4.55 g).

[0332] The above procedure also functioned well when seed crystal of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide Form B (90 mg / 0.015 equivalent, pre-ground with a pestle and mortar) was added to the solution instead.

[0333] As described above, the addition of 0.015 - 0.025 equivalent of ground Form B seed crystal functioned well for the acetonitrile solution. Further modifications of the seed crystal addition procedure were evaluated as described in Table 3 below.

[0334]

Table 3

[0335] (v) Interconversion between Form A / Form B (R)-N-(2-(4-Cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholino-quinoline-4-carboxamide (a 50 / 50 mixture of Form A and Form B, 20 g, 48.61 mmol) was placed in 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 ジャケット = 70 °C), stirred at that temperature for 5 h (300 rpm), and then cooled to 20 °C over 5 h. The suspension was then stirred at 20 °C for an additional 15 h. A very fine and easily stirred suspension was obtained, and no solids adhered to the glass walls of the reactor. The mixture was filtered, and the solid was washed with 25% ethyl acetate in acetone (80 mL). The filter cake was then suction dried for 5 min. Drying was continued overnight under vacuum at 40 °C to obtain 15 g of a yellow product. PXRD and DSC analysis (melting point of 194 °C) were consistent with Form B.

[0336] (vi) Seeding crystallization (methanol) (R)-N-(2-(4-Cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholino-quinoline-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. Then, crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholino-quinoline-4-carboxamide Form B (0.0008 equivalent, pre-ground in a mortar and pestle) was added as seed crystals, and the mixture was stirred at 45 °C for 2 days. The solution was then cooled to 20 °C and held 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 further testing (see Example 7) suggests that (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholino-quinoline-4-carboxamide may be chemically unstable in methanol.

[0337] (vii) Seeding crystallization (acetonitrile / 1-butanol) As the retention time of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholino-quinoline-4-carboxamide in an acetonitrile solution at 70 °C increases, the risk of racemization also increases. To further reduce this risk, (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholino-quinoline-4-carboxamide (12.6 g) was dissolved at 70 °C in a mixture of acetonitrile (31.5 mL) and 1-butanol (58.3 mL), 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. To the solution was added seed crystal of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholino-quinoline-4-carboxamide Form B (4.3 mg, 0.00035 equivalent, pre-ground in a mortar and pestle), and then the mixture was 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 then the product was isolated by filtration under vacuum. The filter cake was washed successively with 1-butanol (12.4 mL) and tert-butyl methyl ether (24.8 mL), and then dried at 40 °C under vacuum. PXRD and DSC analyses were consistent with Form B.

[0338] B. Physical Characterization of Crystalline Form B The characterization of Form B was performed using various techniques including PXRD (Figures 9-11), solid state 13 13C NMR spectroscopy (Figures 2-5), differential scanning calorimetry (DSC) (Figure 12), thermogravimetric analysis (TGA) (Figure 13), and gravimetric vapor sorption (GVS) (Figure 14).

[0339] The PXRD pattern of Figure 9 confirms that Form B is crystalline. Figure 9 shows the PXRD pattern of Form B measured using the transmission mode. In Table 4 below, the selected peaks identified in the PXRD pattern of Figure 9 are listed.

[0340]

Table 4

[0341] Form B shows characteristic 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 (compared to other forms excluding Form A). Form B shows additional 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.

[0342] Form B shows characteristic peaks (i.e., relative intensity peaks of medium intensity or higher) compared to Form A at 18.7 ± 0.2° 2θ and 22.4° 2θ ± 0.2° 2θ. Figure 10 is a comparison of the transmission powder X-ray diffraction patterns for Forms A and B based on the diffractograms shown in Figures 1 and 9. Figure 11 is a comparison of the transmission powder X-ray diffraction patterns for Forms A and B for the region encompassing the characteristic Form B peaks. The diffractogram of Form A is at the top of Figures 10 and 11, and the diffractogram of Form B is at the bottom of Figures 10 and 11.

[0343] As described above for Form A, Figure 2 shows a representative solid state of a sample of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinokynolin-4-carboxamide that is about 95 wt% Form A and 5 wt% Form B. 13 Figure 3 shows a representative solid state 13 13 C NMR spectrum of a sample of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholino-kynolin-4-carboxamide that is about 30 wt% Form A and 70 wt% Form B. Figure 4 shows a representative solid state 13 13The 13C NMR spectrum is shown. Figure 5 shows the solid states of crystalline (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholino-quinoline-4-carboxamide Forms A and B based on the combined spectrum of Figures 2, 3 and 4. 13 This is the analysis of the 13C NMR spectrum. Form B shows 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 characteristic 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 that are characteristic compared to Form A.

[0344] A representative lamp DSC thermogram of Form B is shown in Figure 12. The exothermic events are plotted upward. The endothermic melting shown in Figure 12 has an onset temperature of about 191 °C and a heat enthalpy of about 87 J / g for the endothermic melting. As described above, the DSC values obtained can vary by about ±5 °C depending on the apparatus used, the method by which the sample is prepared, and differences between batches. The DSC thermogram shown in Figure 10 was generated using a DSC Q2000 module. The apparatus was equilibrated at 25 °C and the sample (1 mg - 3 mg) was heated to 230 °C at a rate of 10 °C / min.

[0345] Figure 13 shows a representative TGA thermogram of Form B. Form B shows a weight loss of less than about 0.1% upon heating from about 25 °C to 110 °C, confirming that Form B is an anhydrate.

[0346] Figure 14 shows a representative GVS plot for Form B. Form B showed reversible water uptake of less than about 0.03 wt% at 25 °C ± 0.1 °C from 0% relative humidity to 80% relative humidity. The desorption curve indicates that Form B lost water at a similar rate as the water obtained during sorption with limited hysteresis. No morphological changes were observed by PXRD after the GVS experiment. According to the European Pharmacopoeia (EP) classification, Form B is non-hygroscopic (i.e., < 0.2% weight gain).

[0347] Example 6: Crystal Form 2 Form 2 is a crystalline form of racemic N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-4-carboxamide.

[0348] A. Preparation of (R,S)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-4-carboxamide Crystal Form 2 (R,S)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-4-carboxamide Form 2 can be isolated by slurrying (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-4-carboxamide at high temperature in methanol or a methanol-water mixture for a long time, or by several temperature cycles in chloroform, dioxane or tetrahydrofuran. These methods are described below.

[0349] (i) Slurry at 50 °C (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-4-carboxamide (30 - 40 mg) Form B was slurried in 0.25 mL of a methanol-water mixture with a volume ratio of 84:16 (methanol:water) at 50 °C for 7 days or more. The remaining solid material was isolated and dried before analysis. Analysis confirmed that the isolated crystalline form was Form 2.

[0350] (ii) Temperature cycle (R)-N-(2-(4-Cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholino-quinoline-4-carboxamide amorphous of about 50 mg was slurried in 0.3 mL of a solvent (chloroform, dioxane or tetrahydrofuran). The slurry was subjected to 10 temperature cycles between 5 °C and a temperature ~3 °C lower than the boiling point of each solvent. For the temperature cycle, a ramp of 1 °C / min was used when going up and down. The suspension was centrifuged, the supernatant was decanted, and the solid material was dried before analysis. Analysis confirmed that the isolated crystalline form was Form 2.

[0351] B. Physical Characterization of Crystal Form 2 The characterization of Form 2 was performed using various techniques including PXRD (Figure 15), differential scanning calorimetry (DSC) (Figure 16), and gravimetric vapor sorption (GVS) (Figure 17).

[0352] The PXRD pattern in Figure 15 confirms that Form 2 is crystalline. Figure 15 shows the PXRD pattern of Form 2 measured using a reflection geometry. In Table 5 below, the selected peaks identified in the PXRD pattern of Figure 15 are listed.

[0353]

Table 5

[0354] Form 2 shows characteristic 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 (compared to other crystalline anhydrates). Form 2 shows additional 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°.

[0355] Figure 16 shows a representative lamp DSC thermogram of type 2. The exothermic events are plotted upward. The endothermic melting shown in Figure 19 has an onset temperature of about 201 °C with a heat flow of about 92 J / g for the endothermic melting. As described above, the obtained DSC values can vary by about ±5 °C depending on the apparatus used, the method by which the sample is prepared, and differences between batches.

[0356] Figure 17 shows a representative GVS plot of type 2. Type 2 showed reversible moisture uptake of less than about 0.5 wt% at 25 °C ± 0.1 °C from 0% relative humidity to 80% relative humidity. The desorption curve indicates that type 2 lost moisture at a similar rate to the moisture obtained during sorption with limited hysteresis. According to the European Pharmacopoeia (EP) classification, type 2 is slightly hygroscopic (i.e., ≧0.2% and <2% weight increase).

[0357] Example 7: Crystallization Solvents (Decomposition / Epimerization) Solutions of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholino-quinoline-4-carboxamide (“quinoline”, 1.5 mg / mL) in methanol, ethanol, and acetonitrile were held at ambient temperature and analyzed by supercritical fluid chromatography - mass spectrometry over a maximum of 14 days. New small peaks with masses corresponding to quinoline + 32 and quinoline + 46 were observed in methanol and ethanol, respectively. These peaks are likely derived from compounds formed by the addition of methanol or ethanol to the nitrile of quinoline, i.e., compound 1 and compound 2 and their respective enantiomers. The structures of compound 1 and compound 2 are shown below.

[0358]

Chemical Structure

[0359] Small new peaks corresponding to the masses of the proposed degradation products, 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-morpholino-quinoline-4-carboxamide was observed. The structures of Compound 3 and Compound 4 are shown below.

[0360] [Chemical formula]

[0361] Table 6 below reports the area % data over time measured for the solution held in methanol.

[0362] [Table 6]

[0363] Table 7 below reports the area % data over time measured for the solution held in ethanol.

[0364] [Table 7]

[0365] Table 8 below reports the area % data over time measured for the solution held in acetonitrile. Epimerization of the carboxamide was observed in acetonitrile, but chemical degradation was not observed.

[0366] [Table 8]

[0367] Example 8: Solubility Test A. Solubility Test of Form B Solubility tests were conducted on Form B. Small aliquots of each of the solvents tested were added to accurately weighed samples of Form B (approx. 20 mg) at ambient temperature. The aliquot volume was typically 20 - 200 μL and the total volume was up to 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 approximately 2 mg of Form B.

[0368] The estimated solubility in each of the solvents tested was based on the total solvent used to provide complete dissolution. It should be noted that the actual solubility may vary to some extent from the estimated (i.e., visually detected) solubility due to the use of overly large solvent aliquots, slow dissolution rates, or other factors. The estimated solubilities are reported in Table 9 below.

[0369]

Table 9

[0370] Solvents suitable for cooling crystallization should have a high solubility for the solute and a high potential recovery rate, i.e., the solvent should generally have a high solubility for the solute at high temperatures and a relatively low solubility for the solute at low temperatures (i.e., solubility coefficient at high temperatures). Regarding their solubilities at low temperatures, solvents suitable for cooling crystallization generally have solubilities in the range of approximately 5 mg / mL to approximately 20 mg / mL at 20°C.

[0371] B. FaSSIF Blank Dissolution Medium (Forms A and B) The solubility of Forms A and B in phosphate buffer, pH 6.5 (FaSSIF blank), the dissolution medium, was determined at 25°C after equilibration of the crystal forms in the medium 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 that the crystal form was retained. The solubilities measured for Forms A and B were 0.50 mg / mL and 0.30 mg / mL, respectively.

[0372] C. Acetonitrile Solubility Curve Using a Crystal16 apparatus, a solubility curve with increasing temperature in acetonitrile is created. At least three samples of accurately measured (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide Form B are placed in Crystal 16 vials, acetonitrile (1.0 mL) is added 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 cloud point for each concentration is determined by measuring the turbidity.

[0373] D. Acetonitrile and Methanol Solubility Curves The solubility of Form B in acetonitrile and methanol with increasing temperature was evaluated. (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide Form B (each sample was 14.35 mg - 74.1 mg) and solvent (1.0 mL, either acetonitrile or methanol) were placed in Crystal 16 vials. The contents were stirred at 700 rpm, and the resulting slurry was heated from 25 °C to 70 °C at a constant rate of 0.075 °C / min. The cloud point for each concentration of Form B was determined by measuring the turbidity. The solubility curves created based on the measurement data are shown in Figure 21.

[0374] Example 9: Amorphous (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide A. Preparation of Amorphous (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide Amorphous (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinoquinoline-4-carboxamide can be isolated by various methods including freeze-drying, poor solvent precipitation, flash evaporation, melt quenching, crash precipitation, and vapor diffusion. Some of these methods are shown below.

[0375] (i) Flash evaporation (R)-N-(2-(4-Cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide (25 - 30 mg) is dissolved in hot acetone, chloroform, tetrahydrofuran or dichloromethane (400 - 700 μL). The solution is filtered through a 0.2 μm filter and placed directly into a heating vial on a hot plate at about 120 °C to evaporate the solvent and obtain a glassy solid. The glass residue formed in situ is used without further manipulation.

[0376] (ii) Crash precipitation A saturated solution of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide is prepared in a selected solvent and filtered directly at ambient temperature through a 0.2 μm PTFE filter into a round-bottom flask containing a poor solvent (about 10 volumes).

[0377] (iii) Freeze drying A solution of (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide in acetonitrile / water 1:1 is frozen in liquid nitrogen and dried overnight in a freeze dryer (Christ Alpha 2-4 LDplus).

[0378] (iv) Melt quenching (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide (20 - 100 mg) is added to an HPLC vial and flushed with nitrogen gas. The vial is heated to 200 °C over 5 minutes and then quickly immersed in an ice / water bath to form an amorphous substance.

[0379] B. Physical characterization of amorphous (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide The amorphous (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholino-quinoline-4-carboxamide was characterized by PXRD (Figure 18), differential scanning calorimetry (DSC) (Figure 19), and gravimetric vapor sorption (GVS) (Figure 20). The PXRD pattern of Figure 18 shows that the amorphous compound does not have a regular crystal order.

[0380] Example 10: FAP Inhibition and Binding Assays The hFAP protein used in the examples was either commercially supplied 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). The recombinant hFAP protein was secreted from Sf21 cells in the medium and purified by affinity chromatography (batch mode, Ni excel resin, AKTA, GE Healthcare) and size exclusion chromatography (Superdex200, AKTA, GE Healthcare), concentrated to 19.5 mg / mL, snap frozen in liquid nitrogen, and stored at -80 °C.

[0381] A. hFAP Inhibition Assay Using hFAP enzyme (Proteros, 38 - 760 (PR - 0071)) at 0.24 nM FAC and substrate Ala - Pro - AMC (ARI - 3144) at 20 μM FAC, (R)-N-(2-(4 - cyanothiazolidin - 3 - yl)-2 - oxoethyl)-6 - morpholinokynolin - 4 - carboxamide was tested in a biochemical inhibition assay. 384 - well low - volume black plates (Greiner #784076) were used. 4 μL of 0.48 nM enzyme solution (100 mM Tris HCl, 100 mM NaCl, 0.05% Chaps, pH 7.4) was added to 40 nL of compound (in DMSO) from 50 μM FAC in a 10 - fold, 3 - fold dilution series down to 40 nM. 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 plates were centrifuged at 1000 rpm and incubated in the dark at room temperature for 30 minutes. The plates were read on a PHERAstar® reader at an excitation of 340 nm and emission of 460 nm. The data was analyzed with Genedata Screener®. IC 50 values were determined by plotting % inhibition against log compound concentration and using a one - site dose - response model. The raw data signal was normalized using 0.5% DMSO as the 0% control and reference compound A (i.e., (S)-N-(2-(2 - cyano - 4,4 - difluoropyrrolidin - 1 - yl)-2 - oxoethyl)kynolin - 4 - carboxamide as reported in J.Med.Chem.2014,57,3053) at 50 μM as the 100% inhibitor control. The data is reported in Table 10.

[0382] B. hFAP Inhibition Assay (Tight - Binding Agent) (R)-N-(2-(4-Cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide was tested in a biochemical inhibition assay using 2.4 pM of human fibroblast activation protein alpha (hFAP) enzyme (Proteros, 38-760 (PR-0071)) and 20 μM of substrate Ala-Pro-AMC (ARI-3144). 384-well low-volume black plates (Greiner #784076) were used. 4 μL of 4.8 pM enzyme solution (100 mM Tris HCl, 100 mM NaCl, 0.05% Chaps, pH 7.4) was added to 40 nL of compound (in DMSO) in a 3-fold dilution series from 50 nM to 10 CR. 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 plates were centrifuged at 1000 rpm and incubated in the dark at room temperature for 2.5 hours. The plates were read on a PHERAstar® reader at an excitation of 340 nm and an emission of 460 nm. The data were analyzed with Genedata Screener®. IC 50 values were determined by plotting % inhibition against log compound concentration and using a one-site dose-response model. The raw data signal was normalized using 0.5% DMSO as the 0% control and reference compound A (i.e., (S)-N-(2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)quinoline-4-carboxamide) at 50 μM as the 100% inhibitor control as reported in J. Med. Chem. 2014, 57, 3053. The data are reported in Table 10.

[0383] C.hFAP Binding Assay (R)-N-(2-(4-Cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-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) onto a CMD200M sensor chip (Xantec) was performed using standard amine coupling procedures in immobilization buffer (10 mM HEPES, 150 mM NaCl, 0.05% Tween20, pH 7.4). The surface was washed with 10 mM NaOH, 1 M NaCl, activated with EDC / NHS (GE Healthcare), and subsequently hFAP (in 10 mM acetic acid pH 5.0) was immobilized. Finally, the surface was inactivated with ethanolamine. The level of hFAP immobilization was approximately 4000 - 6000 RU. The reference spot was processed as described, omitting the injection of hFAP. Using single cycle kinetics in running buffer (20 mM TRIS, 150 mM NaCl, 0.05% Tween20, 1% DMSO, pH 7.4), a series of compound concentrations were injected onto the immobilized protein at increasing concentrations (2 - 500 nM). The interaction model was globally fitted to the experimental traces, enabling the determination of k on 、k off and K d . The data are reported in Table 10.

[0384] [Table 10] 1 IC 50 is reported as a single measurement (n = 1) or as the geometric mean of multiple measurements (n = 2 - 3). 2 IC 50 is reported as a single measurement (n = 1) or as the geometric mean of multiple measurements (n = 2 - 6). 3 K d is reported as a single measurement (n = 1) or as the geometric mean of multiple measurements (n = 2 - 4). k (on) and k (off)It is reported as the average of a single measurement (n = 1) or multiple measurements (n = 2 - 4). NV means not valid.

[0385] D.FAP Plasma Inhibition Assay Plasma (anticoagulant K2EDTA) was used as the enzyme source: human plasma (pooled from AZ Biobank), mouse plasma (AZ AST Biobank), and cynomolgus monkey plasma (BioIVT, #NHP00PLK2FNN, lot CYN222895). A 384-well black fluotrack PS plate (Greiner 781076) was used. 20 μL of diluted plasma (cynomolgus monkey and human plasma diluted 1:40, mouse plasma diluted 1:67) in buffer (PBS, 0.1% BSA) was added to 0.6 μL of the compound (in DMSO). The compound was tested using a 3-fold dilution series from 500 nM FAC to 10CR. Two replicates for each assay point were performed on the same plate. Fluorescence blanks 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 obtain 75 μM FAC. The plate was incubated in the dark at room temperature for 40 minutes. The plate was read on a Beckman Paradigm® reader at an excitation of 360 nm and an emission of 465 nm. The data were analyzed in Excel (IDBS XLfit Add-In) using a one-site dose-response model (4-parameter logistic fit). IC 50 values were determined by plotting % inhibition against log compound concentration. The raw data signal was normalized using 1.5% DMSO in diluted plasma as the 0% control and 1.5% DMSO in buffer (without plasma) as the 100% inhibitor control. The data are reported in Table 11.

[0386]

Table 11

[0387] Example 11: hPrep Inhibition Assay Using prolyl endopeptidase, 0.6 nM FAC of prolyl oligopeptidase (hPREP) enzyme (R&D Systems, 4308-SE) and 50 μM FAC of substrate Z-Gly-Pro-amino-methylcoumarin (Bachem, I-1145), (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholinquinoline-4-carboxamide was tested in a biochemical inhibition assay. 384-well low-volume black plates (Greiner #784076) were used. 4 μL of 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 of the compound (in DMSO) in a 10CR, 3-fold dilution series from 50 μM FAC. The plates were incubated in the dark at room temperature 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 plates were centrifuged at 1000 rpm and incubated in the dark at room temperature for 20 minutes. The plates were read on a PHERAstar® reader at an excitation of 340 nm and an emission of 460 nm. The data was analyzed with Genedata Screener®. IC 50Values were determined by plotting % inhibition against log compound concentration and using a one-site dose-response model. Raw data signals were normalized using 0.5% DMSO as the 0% control and 50 μM of reference compound B (i.e., (R)-N-(2-(4-cyanothiazolidin-3-yl)-2-oxoethyl)-7-methylquinoline-4-carboxamide) as the 100% inhibitor control. Data are reported in Table 12.

[0388]

Table 12

[0389] Example 12: hDPP Inhibition Assay A. hDPP7 Inhibition Assay (R)-N-(2-(4-Cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-4-carboxamide was tested in a biochemical inhibition assay using 15 nM FAC of human dipeptidyl peptidase 7 (hDPP7) enzyme (BPS Bioscience, #80070) and 5 μM FAC of substrate Ala-Pro-amino-methylcoumarin (BPS Bioscience, #80305). The enzyme reaction was performed in duplicate for 30 minutes at room temperature in 50 μL of DPP assay buffer (BPS Bioscience, #80300). A 10CR, 3-fold dilution series of the compound solution (in DMSO) was prepared in assay buffer at 10-fold higher than the final concentration, and 5 μL of the dilution was added to 50 μL of the reaction such that the highest compound concentration was 100 μM FAC and the concentration of DMSO was 1% in all wells. The plate was read on a Tecan Infinite M1000 microplate reader at an excitation of 340 nm and emission of 460 nm. The data were analyzed with Graph Pad Prism. IC 50 values were determined by plotting % inhibition against log compound concentration and using a one-site dose-response model. The raw data signal was normalized using 1% DMSO as the 0% control and enzyme-free as the 100% inhibitor control. The data are reported in Table 13.

[0390] B. hDPP8 Inhibition Assay (R)-N-(2-(4-Cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-4-carboxamide was tested in a biochemical inhibition assay using 1.5 nM FAC human dipeptidyl peptidase 8 (hDPP8) enzyme (BPS Bioscience, #80080) and 5 μM FAC substrate Ala-Pro-amino-methylcoumarin (BPS Bioscience, #80305). Enzyme reactions were performed in duplicate for 30 minutes at room temperature in 50 μL of DPP assay buffer (BPS Bioscience, #80300). A 10CR, 3-fold dilution series of the compound solution (in DMSO) was prepared in assay buffer at 10-fold higher than the final concentration, and 5 μL of the dilution was added to 50 μL of the reaction such that the highest compound concentration was 100 μM FAC and the concentration of DMSO was 1% in all wells. The plate was read on a Tecan Infinite M1000 microplate reader at an excitation of 340 nm and emission of 460 nm. Data were analyzed with Graph Pad Prism. IC 50 values were determined by plotting % inhibition versus log compound concentration and using a one-site dose-response model. The raw data signal was normalized using 1% DMSO as the 0% control and no enzyme as the 100% inhibitor control. Data are reported in Table 13.

[0391] C.hDPP9 Inhibition Assay (R)-N-(2-(4-Cyanothiazolidin-3-yl)-2-oxoethyl)-6-morpholin-4-ylquinoline-4-carboxamide was tested in a biochemical inhibition assay using 0.4 nM FAC human dipeptidyl peptidase 9 (hDPP9) enzyme (BPS Bioscience, #80090) and 5 μM FAC substrate Ala-Pro-amino-methylcoumarin (BPS Bioscience, #80305). The enzyme reaction was performed in duplicate for 30 minutes at room temperature in 50 μL of DPP assay buffer (BPS Bioscience, #80300). A 10 CR, 3-fold dilution series of the compound solution (in DMSO) was prepared in assay buffer 10-fold higher than the final concentration, and 5 μL of the dilution was added to 50 μL of the reaction such that the highest compound concentration was 100 μM FAC and the concentration of DMSO was 1% in all wells. The plate was read on a Tecan Infinite M1000 microplate reader at an excitation of 340 nm and an emission of 460 nm. The data were analyzed using Graph Pad Prism. IC 50 values were determined by plotting % inhibition versus log compound concentration and using a one-site dose-response model. The raw data signal was normalized using 1% DMSO as the 0% control and no enzyme as the 100% inhibitor control. The data are reported in Table 13.

[0392]

Table 13

[0393] The above specific embodiments and examples have been described, but these embodiments and examples are presented only as examples and are not intended to limit the scope of the invention. Changes and modifications can be made by those skilled in the art without departing from the broader aspects of the present disclosure as defined in the following claims. For example, any of the embodiments described herein can be combined with any other suitable embodiments described herein to provide additional embodiments.

Claims

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

3. The crystalline morphology according to claim 2, characterized by a transmitted X-ray powder diffraction pattern including peaks at 18.7 ± 0.2°²θ and 22.4°²θ ± 0.2°²θ.

4. The crystal morphology according to claim 3, wherein the transmitted X-ray powder diffraction pattern further includes 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. Solid state containing 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 crystalline form according to claim 2, further characterized by a 13C NMR spectrum.

6. The solid state １３ The crystalline form according to claim 5, wherein the 13C NMR spectrum includes peaks at 49.9 ± 0.2 ppm and 46.6 ± 0.2 ppm.

7. The crystalline morphology according to claim 2, further characterized by a differential scanning calorimetry curve that includes endothermic melting starting at approximately 185°C to approximately 200°C.

8. The crystal morphology according to claim 3, further characterized by a thermogravimetric thermogram showing that the crystal morphology exhibits a weight loss of less than 0.5% by weight at temperatures between approximately 25°C and approximately 110°C.

9. The crystal morphology according to claim 3, further characterized by a gravimetric vapor sorption plot showing that the crystal morphology exhibits reversible moisture uptake of less than 0.5% by weight at 25°C ± 0.1°C and approximately 0% relative humidity to approximately 80% relative humidity.

10. The crystalline form according to claim 3, which is a crystalline anhydride.

11. The transmitted X-ray powder diffraction pattern includes 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. The crystal morphology according to claim 2, wherein the transmitted X-ray powder diffraction pattern does not include peaks of medium or higher relative intensity at 18.7 ± 0.2°²θ and 22.4°²θ ± 0.2°²θ.

12. The crystal morphology according to claim 11, wherein the transmitted X-ray powder diffraction pattern further includes 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. Solid state containing 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 crystalline form according to claim 2, further characterized by a 13C NMR spectrum.

14. The solid state １３ The crystalline form according to claim 13, wherein the 13C NMR spectrum includes peaks at 166.1 ± 0.2 ppm, 117.9 ± 0.2 ppm, and 108.6 ± 0.2 ppm.

15. The crystalline morphology according to claim 2, further characterized by a differential scanning calorimetry curve including endothermic melting with a starting temperature of approximately 165°C to approximately 180°C.

16. The crystal morphology according to claim 11, further characterized by a thermogravimetric thermogram showing that the crystal morphology exhibits a weight loss of less than 0.5% by weight at temperatures between approximately 25°C and approximately 110°C.

17. The crystal morphology according to claim 11, further characterized by a gravimetric vapor sorption plot showing that the crystal morphology exhibits reversible moisture uptake of less than 0.5% by weight at 25°C ± 0.1°C to about 80% relative humidity.

18. The crystalline form according to claim 11, which is a crystalline anhydride.

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

20. The crystal form described in claim 3, The crystal form described in claim 11, and Crystal form according to claim 19 A composition comprising at least two crystalline forms selected from the group consisting of the following.

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

22. The pharmaceutical composition according to claim 21 for treating or preventing a FAP-mediated condition.

Images