Crystalline forms of TEAD inhibitor

Abstract

The specification relates to physical forms of a TEAD inhibitor 5-(((3S,5R)-5-hydroxytetrahydro-2H- pyran-3-yl)amino)-3-methyl-8-(4-(trifluoromethyl)phenyl)pyrido[4,3-d]pyrimidin-4(3H)-one, to pharmaceutical compositions containing them and their use in therapy.

Description

[0001] CRYSTALLINE FORMS OF TEAD INHIBITOR

[0002] Cross-Reference to Related Patent Application

[0003] This specification claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 593,253 (filed October 26 2023). The entire text of the above-referenced patent application is incorporated by reference into this specification.

[0004] Field

[0005] This specification relates to crystalline forms of 5-(((3S,5R)-5-hydroxytetrahydro-2H-pyran-3- yl)amino)-3-methyl-8-(4-(trifluoromethyl)phenyl)pyrido[4,3-d]pyrimidin-4(3H)-one (Compound (I)), to pharmaceutical compositions containing them and their use in therapy. Compound (I) has been discovered to be an inhibitor of TEAD and consequently has potential utility as a medicine for the treatment of cancer.

[0006] Introduction

[0007] The Hippo pathway is a highly conserved signaling pathway that controls organ size and tissue maintenance through the regulation of gene expression programs involved in cell proliferation, survival, and differentiation (Dong et al., Cell 2007, 1120-33; Ma et al., Ann Rev Biochem 2018, 577- 604 and references therein). Hippo ultimately regulates the transcription coactivators Yes-associated protein (YAP) and transcriptional coactivator with PDZ-binding motif (TAZ) which bind to DNA-bound Transcriptional Enhanced Associate Domain proteins (TEAD1-4) to form bipartite transcription complexes that activate TEAD-dependent gene expression. The core of the Hippo pathway consists of a tightly regulated kinase signaling cascade. When Hippo signaling is active, the kinases LATS1 / 2 phosphorylate YAP / TAZ which causes these proteins to be sequestered in the cytoplasm or degraded by the proteasome. When Hippo signaling is inactive, LATS1 / 2 are inactivated resulting in YAP / TAZ to be dephosphorylated and subsequently translocated into the nucleus to interact with and activate TEAD-dependent transcription (Meng et al., Genes&Dev 2016, 1-17).

[0008] Hippo signaling is a well-established tumor suppressor pathway and data from The Cancer Genome Atlas show that the Hippo pathway is one of eight signaling pathways that are frequently altered in human cancer (Sanchez-Vega et al., Cell 2018, 321-337). Both genetic and epigenetic alterations of Hippo components can result in aberrant activation of YAP / TAZ and TEAD-dependent transcription and have been implicated in several human malignancies (Wang et al., 2018, 1304-1317). NF2 (aka Merlin) is encoded by the neurofibromatosis type 2 gene and is a key upstream regulator of the Hippo core kinase cascade consisting of STE20-like protein kinase 1 (STK3, aka MST2, and STK4, aka MST1), the large tumor suppressors (LATS1 and LATS2), and adaptor proteins Salvador homolog 1 (SAV1) and MOB kinase activators (MOB1A / MOB1B) (Tapon et al., Cell 2002, 467-478). Loss of function mutations or deletions in pathway components have been reported in several cancer types including mesothelioma, breast, liver, lung, prostate, gastric, and colorectal tumors (Poma et al., Scientific Reports 2018, volume 8, Article number: 10623; Zancanato et al., Cancer Cell 2016, 783- 803 and references therein).

[0009] Because several Hippo pathway components are tumor suppressors where dysfunction results in aberrant TEAD-dependent transcription, targeting TEAD offers a potential opportunity for therapy.

[0010] 5-(((3S,5R)-5-hydroxytetrahydro-2H-pyran-3-yl)amino)-3-methyl-8-(4- (trifluoromethyl)phenyl)pyrido[4,3-d]pyrimidin-4(3H)-one,

[0011] (Compound (I)) is disclosed in PCT / EP2023 / 061101 as an inhibitor of TEAD, the contents of which are incorporated by reference. PCT / EP2023 / 061101 discloses that Compound (I) was isolated as a white solid. This solid was found to be crystalline Form A, as characterised herein.

[0012] In order to study the therapeutic potential of this compound it is desirable to have solid forms of the compound with suitable properties for pharmaceutical development. This application describes new crystalline forms of Compound (I) (Forms B and C).

[0013] In the formulation of drug substances, it is beneficial for the drug substance (active compound) to be in a form in which it can be conveniently handled and processed. The chemical stability and the physical stability of the active compound may be important factors in determining the suitability of a solid form for use in the development of pharmaceutical formulations. It is also beneficial for the active compound, and formulations containing it, to be capable of being effectively stored over appreciable periods of time, without exhibiting any significant change in the physico-chemical characteristics (e.g. chemical composition, density, hygroscopicity and solubility) of the active compound. General Description

[0014] In a first aspect there is provided a crystalline form of 5-(((3S,5R)-5-hydroxytetrahydro-2H-pyran-3- yl)arnino)-3-methyl-8-(4-(trifluorornethyl)phenyl)pyrido[4,3-d]pyrirnidin-4(3H)-one (Compound (I)) that is Form C.

[0015] "Form C" has an X-ray powder diffraction pattern with specific peaks at about 2-theta = 6.4 and 9.7°, when measured using CuKa radiation.

[0016] "Form C" has an X-ray powder diffraction pattern substantially as shown in Figure 7 , when measured using CuKa radiation.

[0017] In a further aspect there is provided a pharmaceutical composition comprising the crystalline form of Compound (I) Form C, and a pharmaceutically acceptable excipient.

[0018] In a further aspect there is provided the crystalline form of Compound (I) Form C, for use in therapy.

[0019] In a further aspect there is provided the crystalline form of Compound (I) Form C, for use in the treatment of cancer.

[0020] In a further aspect there is provided the use of the crystalline form of Compound (I) Form C in the manufacture of a medicament.

[0021] In a further aspect there is provided the use of the crystalline form of Compound (I) Form C, in the manufacture of a medicament for the treatment of cancer.

[0022] In a further aspect there is provided a method of treating liver disease in a patient comprising administering to the patient an effective amount of the crystalline form of Compound (I) Form C.

[0023] In a further aspect there is provided a crystalline form of 5-(((3S,5R)-5-hydroxytetrahydro-2H-pyran- 3-yl)amino)-3-methyl-8-(4-(trifluoromethyl)phenyl)pyrido[4,3-c / ]pyrimidin-4(3H)-one (Compound (I)) that is Form B.

[0024] "Form B" has an X-ray powder diffraction pattern with specific peaks at about 2-theta = 5.4° and 10.9°, when measured using CuKa radiation.

[0025] "Form B" has an X-ray powder diffraction pattern substantially as shown in Figure 4, when measured using CuKa radiation.

[0026] In a further aspect there is provided a pharmaceutical composition comprising the crystalline form of Compound (I) Form B, and a pharmaceutically acceptable excipient.

[0027] In a further aspect there is provided the crystalline form of Compound (I) Form B, for use in therapy. In a further aspect there is provided the crystalline form of Compound (I) Form B, for use in the treatment of cancer.

[0028] In a further aspect there is provided the use of the crystalline form of Compound (I) Form B in the manufacture of a medicament.

[0029] In a further aspect there is provided the use of the crystalline form of Compound (I) Form B, in the manufacture of a medicament for the treatment of cancer.

[0030] In a further aspect there is provided a method of treating liver disease in a patient comprising administering to the patient an effective amount of the crystalline form of Compound (I) Form B.

[0031] Description of Figures

[0032] Embodiments and experiments illustrating the principles of the disclosure will now be discussed with reference to the accompanying figures in which:

[0033] Figure 1 shows a powder X-ray diffraction diagram of Compound (I) Form A.

[0034] Figure 2 shows differential scanning calorimetry (DSC) traces of Compound (I) Form A.

[0035] Figure 3 shows thermogravimetric analysis (TGA) traces of Compound (I) Form A.

[0036] Figure 4 shows a powder X-ray diffraction diagram of Compound (I) Form B.

[0037] Figure 5 shows differential scanning calorimetry (DSC) traces of Compound (I) Form B.

[0038] Figure 6 shows thermogravimetric analysis (TGA) traces of Compound (I) Form B.

[0039] Figure 7 shows a powder X-ray diffraction diagram of Compound (I) Form C.

[0040] Figure 8 shows differential scanning calorimetry (DSC) traces of Compound (I) Form C.

[0041] Figure 9 shows thermogravimetric analysis (TGA) traces of Compound (I) Form C.

[0042] Detailed Description

[0043] Form A

[0044] Compound (I) Form A, a crystalline form of 5-(((3S,5R)-5-hydroxytetrahydro-2H-pyran-3-yl)amino)-3- methyl-8-(4-(trifluorornethyl)phenyl)pyrido[4,3-d]pyrimidin-4(3H)-one, has an X-ray powder diffraction pattern substantially as shown in Figure 1, when measured using CuKa radiation. Form A is believed to be a monohydrate of Compound (I). The ten most prominent peaks are shown in Table l.s

[0045] Table 1 Ten most prominent peaks of X-ray powder diffraction pattern of Compound (I) Form A. According to the present specification there is provided Compound (I) Form A which has an X-ray powder diffraction pattern with at least one specific peak at about 2-theta = 5.9°.

[0046] According to the present specification there is provided Compound (I) Form A, which has an X-ray powder diffraction pattern with at least one specific peak at about 2-theta = 11.7°.

[0047] In embodiments there is provided Compound (I) Form A, which has an X-ray powder diffraction pattern with specific peaks at about 2-theta = 5.9° and 11.7°.

[0048] In embodiments there is provided Compound (I) Form A, which has an X-ray powder diffraction pattern with specific peaks at about 2-theta = 5.9, 7.2, 11.0, 11.7, and 17.6°.

[0049] In embodiments there is provided Compound (I) Form A, which has an X-ray powder diffraction pattern with specific peaks at about 2-theta = 5.9, 7.2, 11.0, 11.7, 13.4, 16.0, 17.6, 18.8, 23.5, and 24.3°.

[0050] In embodiments there is provided Compound (I) Form A, which has an X-ray powder diffraction pattern substantially the same as the X-ray powder diffraction pattern shown in Figure 1.

[0051] According to the present specification there is provided Compound (I) Form A, which has an X-ray powder diffraction pattern with at least one specific peak at 2-theta = 5.9° plus or minus 0.2° 2-theta. According to the present specification there is provided Compound (I) Form A, which has an X-ray powder diffraction pattern with at least one specific peak at 2-theta = 11.7° plus or minus 0.2° 2-theta.

[0052] In embodiments there is provided Compound (I) Form A, which has an X-ray powder diffraction pattern with specific peaks at 2-theta = 5.9° and 11. T , wherein said values may be plus or minus 0.2° 2-theta.

[0053] In embodiments there is provided Compound (I) Form A, which has an X-ray powder diffraction pattern with specific peaks at 2-theta = 5.9, 7.2, 11.0, 11.7, and 17.6°, wherein said values may be plus or minus 0.2° 2-theta.

[0054] In embodiments there is provided Compound (I) Form A, which has an X-ray powder diffraction pattern with specific peaks at 2-theta = 5.9, 7.2, 11.0, 11.7, 13.4, 16.0, 17.6, 18.8, 23.5, and 24.3°, wherein said values may be plus or minus 0.2° 2-theta.

[0055] When it is stated that the present specification relates to Compound (I) Form A, the degree of crystallinity is greater than about 60%, greater than about 80%, greater than about 90% or greater than about 95%. In embodiments, the degree of crystallinity is greater than about 98%.

[0056] The Compound (I) Form A provides X-ray powder diffraction patterns substantially the same as the X- ray powder diffraction patterns shown in Figure 1 and has substantially the ten most prominent peaks (angle 2-theta values) shown in Table 1. It will be understood that the 2-theta values of the X-ray powder diffraction pattern may vary slightly from one machine to another or from one sample to another, and so the values quoted are not to be construed as absolute.

[0057] It is known that an X-ray powder diffraction pattern may be obtained which has one or more measurement errors depending on measurement conditions (such as equipment or machine used). In particular, it is generally known that intensities in an X-ray powder diffraction pattern may fluctuate depending on measurement conditions. Therefore it should be understood that the Compound (I) Form A of the present specification is not limited to the crystals that provide X-ray powder diffraction patterns identical to the X-ray powder diffraction pattern shown in Figure 1, and any crystals providing X-ray powder diffraction patterns substantially the same as those shown in Figure 1 fall within the scope of the present specification. A person skilled in the art of X-ray powder diffraction is able to judge the substantial identity of X-ray powder diffraction patterns.

[0058] Form B

[0059] Compound (I) Form B, a crystalline form of 5-(((3S,5R)-5-hydroxytetrahydro-2H-pyran-3-yl)amino)-3- methyl-8-(4-(trifluoromethyl)phenyl)pyrido[4,3-d]pyrimidin-4(3H)-one, having an X-ray powder diffraction pattern substantially as shown in Figure 4 when measured using CuKa radiation. Form B is believed to be a dihydrate of Compound (I). The ten most prominent peaks are shown in Table 2:

[0060] Table 2 Ten most prominent peaks of X-ray powder diffraction pattern of Compound (I) Form B.

[0061] According to the present specification there is provided Compound (I) Form B which has an X-ray powder diffraction pattern with a specific peak at about 2-theta = 5.4°.

[0062] According to the present specification there is provided Compound (I) Form B which has an X-ray powder diffraction pattern with a specific peak at about 2-theta = 10.9°.

[0063] In embodiments there is provided Compound (I) Form B, which has an X-ray powder diffraction pattern with specific peaks at about 2-theta = 5.4° and 10.9°.

[0064] In embodiments there is provided Compound (I) Form B, which has an X-ray powder diffraction pattern with specific peaks at about 2-theta = 5.4, 10.9, 14.2, 19.8, and 23.7°.

[0065] In embodiments there is provided Compound (I) Form B, which has an X-ray powder diffraction pattern with specific peaks at about 2-theta = 5.4, 10.9, 13.4, 14.2, 18.9, 19.3, 19.8, 22.4, 22.9, and 23.7°.

[0066] According to the present specification there is provided Compound (I) Form B which has an X-ray powder diffraction pattern substantially the same as the X-ray powder diffraction pattern shown in Figure 4.

[0067] According to the present specification there is provided Compound (I) Form B which has an X-ray powder diffraction pattern with a specific peak at 2-theta = 5.4° plus or minus 0.2° 2-theta. According to the present specification there is provided Compound (I) Form B which has an X-ray powder diffraction pattern with a specific peak at 2-theta = 10.9° plus or minus 0.2° 2-theta.

[0068] In embodiments there is provided Compound (I) Form B, having an X-ray powder diffraction pattern with specific peaks at 2-theta = 5.4° and 10.9°, wherein said values may be plus or minus 0.2° 2-theta.

[0069] In embodiments there is provided Compound (I) Form B, having an X-ray powder diffraction pattern with specific peaks at about 2-theta = 5.4, 10.9, 14.2, 19.8, and 23.7°, wherein said values may be plus or minus 0.2° 2-theta.

[0070] In embodiments there is provided Compound (I) Form B, having an X-ray powder diffraction pattern with specific peaks at about 2-theta = 5.4, 10.9, 13.4, 14.2, 18.9, 19.3, 19.8, 22.4, 22.9, and 23.7°, wherein said values may be plus or minus 0.2° 2-theta.

[0071] When it is stated that the present specification relates to a Compound (I) Form B, the degree of crystallinity is greater than about 60%, greater than about 80%, greater than about 90% and more greater than about 95%. In embodiments, the degree of crystallinity is greater than about 98%.

[0072] The Compound (I) Form B provides X-ray powder diffraction patterns substantially the same as the X- ray powder diffraction patterns shown in Figure 4 and has substantially the ten most prominent peaks (angle 2-theta values) shown in Table 2. It will be understood that the 2-theta values of the X-ray powder diffraction pattern may vary slightly from one machine to another or from one sample to another, and so the values quoted are not to be construed as absolute.

[0073] It is known that an X-ray powder diffraction pattern may be obtained which has one or more measurement errors depending on measurement conditions (such as equipment or machine used). In particular, it is generally known that intensities in an X-ray powder diffraction pattern may fluctuate depending on measurement conditions. Therefore it should be understood that the Compound (I) Form B of the present specification is not limited to the crystals that provide X-ray powder diffraction patterns identical to the X-ray powder diffraction pattern shown in Figure 4, and any crystals providing X-ray powder diffraction patterns substantially the same as those shown in Figure 4 fall within the scope of the present specification. A person skilled in the art of X-ray powder diffraction is able to judge the substantial identity of X-ray powder diffraction patterns.

[0074] FORM C

[0075] Compound (I) Form C, a crystalline form of 5-(((3S,5R)-5-hydroxytetrahydro-2H-pyran-3-yl)amino)-3- methyl-8-(4-(trifluoromethyl)phenyl)pyrido[4,3-d]pyrimidin-4(3H)-one, has an X-ray powder diffraction pattern substantially as shown in Figure 7 when measured using CuKa radiation. Form C is believed to be a monohydrate of Compound (I). The ten most prominent peaks are shown in Table 3:

[0076] Table 3 Ten most prominent peaks of X-ray powder diffraction pattern of Compound (I) Form C. According to the present specification there is provided Compound (I) Form C, which has an X-ray powder diffraction pattern with a specific peak at about 2-theta = 6.4°.

[0077] According to the present specification there is provided Compound (I) Form C, which has an X-ray powder diffraction pattern with a specific peak at about 2-theta = 9.7°.

[0078] In embodiments there is provided Compound (I) Form C, which has an X-ray powder diffraction pattern with specific peaks at about 2-theta = 6.4 and 9.7°.

[0079] In embodiments there is provided Compound (I) Form C, which has an X-ray powder diffraction pattern with specific peaks at about 2-theta = 6.4, 9.7, 16.9, 20.3, and 21.0°.

[0080] In embodiments there is provided Compound (I) Form C, which has an X-ray powder diffraction pattern with specific peaks at about 2-theta = 6.4, 9.7, 13.3, 16.1, 16.9, 19.3, 20.3, 21.0, 22.2, and 24.1°.

[0081] According to the present specification there is provided Compound (I) Form C, which has an X-ray powder diffraction pattern substantially the same as the X-ray powder diffraction pattern shown in Figure 7. According to the present specification there is provided a crystalline form, Compound (I) Form C, which has an X-ray powder diffraction pattern with a specific peak at 2-theta = 6.4° plus or minus 0.2° 2-theta.

[0082] According to the present specification there is provided a crystalline form, Compound (I) Form C, which has an X-ray powder diffraction pattern with a specific peak at 2-theta = 9.7° plus or minus 0.2° 2-theta.

[0083] In embodiments there is provided Compound (I) Form C, having an X-ray powder diffraction pattern with specific peaks at 2-theta = 6.4° and 9.7°, wherein said values may be plus or minus 0.2° 2-theta.

[0084] In embodiments there is provided Compound (I) Form C, having an X-ray powder diffraction pattern with specific peaks at about 2-theta = 6.4, 9.7, 16.9, 20.3, and 21.0°, wherein said values may be plus or minus 0.2° 2-theta.

[0085] In embodiments there is provided Compound (I) Form C, having an X-ray powder diffraction pattern with specific peaks at about 2-theta = 6.4, 9.7, 13.3, 16.1, 16.9, 19.3, 20.3, 21.0, 22.2, and 24.1°, wherein said values may be plus or minus 0.2° 2-theta.

[0086] When it is stated that the present specification relates to a Compound (I) Form C, the degree of crystallinity is greater than about 60%, greater than about 80%, greater than about 90%, or greater than about 95%. In embodiments, the degree of crystallinity is greater than about 98%.

[0087] The Compound (I) Form C provides X-ray powder diffraction patterns substantially the same as the X- ray powder diffraction patterns shown in Figure 7 and has substantially the ten most prominent peaks (angle 2-theta values) shown in Table 3. It will be understood that the 2-theta values of the X-ray powder diffraction pattern may vary slightly from one machine to another or from one sample to another, and so the values quoted are not to be construed as absolute.

[0088] It is known that an X-ray powder diffraction pattern may be obtained which has one or more measurement errors depending on measurement conditions (such as equipment or machine used). In particular, it is generally known that intensities in an X-ray powder diffraction pattern may fluctuate depending on measurement conditions. Therefore it should be understood that the Compound (I) Form C of the present specification is not limited to the crystals that provide X-ray powder diffraction patterns identical to the X-ray powder diffraction pattern shown in Figure 7, and any crystals providing X-ray powder diffraction patterns substantially the same as those shown in Figure 7 fall within the scope of the present specification. A person skilled in the art of X-ray powder diffraction is able to judge the substantial identity of X-ray powder diffraction patterns. X-ray Powder Diffraction Analysis

[0089] The X-ray powder diffraction analysis was performed according to standard methods, which can be found in e.g. Kitaigorodsky, A.L (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.

[0090] Persons skilled in the art of X-ray powder diffraction will realize that the relative intensity of peaks can be affected by, for example, grains above 30 microns in size and non-unitary aspect ratios, which may affect analysis of samples. The skilled person will also realize that the position of reflections can be affected by the precise height at which the sample sits in the diffractometer and the zero calibration of the diffractometer. The surface planarity of the sample may also have a small effect. Hence the diffraction pattern data presented are not to be taken as absolute values. (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).

[0091] Generally, a measurement error of a diffraction angle in an X-ray powder diffractogram is about 5% or less, in particular plus or minus 0.2° 2-theta, and such degree of a measurement error should be taken into account when considering the X-ray powder diffraction patterns in Figure 1, Figure 4 and Figure 7 and when reading Table 1, Table 2 and Table 3. Furthermore, it should be understood that intensities may fluctuate depending on experimental conditions and sample preparation (preferred orientation). Definition of relative intensity is described in Table 3.

[0092] X-ray powder diffraction data was measured with Corundum as an internal reference. The X-ray powder diffraction (referred to herein as XRPD) pattern was determined by mounting a sample on a zero background holder single silicon crystal and spreading out the sample into a thin layer.

[0093] The powder X-ray diffraction was recorded with a theta-two theta scan axis and in one dimensional scan with a Bruker D8 diffractometer (wavelength of X-rays 1.5418 A nickel-f iltered Cu Karadiation, 40 kV, 30 mA) equipped with D / Tex Ultra detector. Automatic variable anti scattering slits were used and the samples were rotated at 30 revolution per minute during measurement. Samples were scanned from 5 - 50° 2-theta using a 0.01° and l° / min step width and scan speed respectively.

[0094] The X-ray powder diffraction analysis was performed according to standard methods, which can be found in e.g. Kitaigorodsky, A.L (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 for polycrystalline and amorphous materials, John Wiley, New York, London.

[0095] Table 1 Definition of relative intensity

[0096] * The relative intensities are derived from diffractograms measured with fixed slits

[0097] A person skilled in the art understands that the value or range of values observed in a particular compound's DSC Thermogram will show variation between batches of different purities. Therefore, whilst for one compound the range may be small, for others the range may be quite large. Generally, a measurement error of a diffraction angle in DSC thermal events is approximately plus or minus 5°C, and such degree of a measurement error should be taken into account when considering the DSC data included herein.

[0098] Crystallisation of the desired form in a process described herein may be aided by seeding with crystals of the desired form. The seed crystals may be obtained using one of the methods described in the Examples. The use of seeding is particularly advantageous in larger-scale manufacture.

[0099] In the preceding paragraphs defining the X-ray powder diffraction peaks for the crystalline form of Compound (I), the term "about=" is used in the expression" ... at about 2-theta = ... "to indicate that the precise position of peaks (i.e. the recited 2-theta angle values) should not be construed as being absolute values because, as will be appreciated by those skilled in the art, the precise position of the peaks may vary slightly between one measurement apparatus and another, from one sample to another, or as a result of slight variations in measurement conditions utilised. It is also stated in the preceding paragraphs that the crystalline form of Compound (I) provide X-ray powder diffraction patterns 'substantially' the same as the X-ray powder diffraction patterns shown in Figure 1, Figure 4 and Figure 7 has substantially the most prominent peaks (2-theta angle values) shown in Table 1, Table 2 and Table 3 respectively. It is to be understood that the use of the term 'substantially' in this context is also intended to indicate that the 2-theta angle values of the X-ray powder diffraction patterns may vary slightly from one apparatus to another, from one sample to another, or as a result of slight variations in measurement conditions utilised, so the peak positions shown in the Figures or quoted in the Tables are again not to be construed as absolute values.

[0100] The person skilled in the art of X-ray powder diffraction will realize that the relative intensity of peaks can be affected by, for example, grains above approximately 30 micrometer in size and non-unitary aspect ratios which may affect analysis of samples. Furthermore, it should be understood that intensities may fluctuate depending on experimental conditions and sample preparation such as preferred orientation of the particles in the sample. The use of automatic or fixed divergence slits will also influence the relative intensity calculations. A person skilled in the art can handle such effects when comparing diffraction patterns.

[0101] The person skilled in the art of X-ray powder diffraction will also realize that due to difference in sample heights and errors in the calibration of the detector position, a small shift in the 2-theta positions could occur. Generally, a difference of ±0.1° from the given value are to be considered correct.

[0102] Crystalline forms of Compound (I) described herein may also be characterised and / or distinguished from other physical forms using other suitable analytical techniques, for example NIR spectroscopy or solid-state nuclear magnetic resonance spectroscopy.

[0103] The chemical structure of Compound (I) forms described herein can be confirmed by routine methods for example proton nuclear magnetic resonance (NMR) analysis.

[0104] Crystalline forms of Compound (I) may be prepared as described in the Examples hereinafter.

[0105] Where "crystalline form of Compound (I)" is mentioned hereafter, this includes Form A, Form B and Form C of Compound (I).

[0106] Medical and pharmaceutical use

[0107] In one aspect of the present specification there is provided a crystalline form of Compound (I), for use in therapy.

[0108] In one aspect of the present specification there is provided a crystalline form of Compound (I), for use in the treatment of cancer.

[0109] Where "cancer" is mentioned, this includes both non-metastatic cancer and also metastatic cancer, such that treating cancer involves treatment of both primary tumours and also tumour metastases.

[0110] The term "therapy" is intended to have its normal meaning of dealing with a disease in order to entirely or partially relieve one, some or all of its symptoms, or to correct or compensate for the underlying pathology. The term "therapy" also includes "prophylaxis" unless there are specific indications to the contrary. The terms "therapeutic" and "therapeutically" should be interpreted in a corresponding manner.

[0111] The term "prophylaxis" is intended to have its normal meaning and includes primary prophylaxis to prevent the development of the disease and secondary prophylaxis whereby the disease has already developed and the patient is temporarily or permanently protected against exacerbation or worsening of the disease or the development of new symptoms associated with the disease.

[0112] The term "treatment" is used synonymously with "therapy". Similarly the term "treat" can be regarded as "applying therapy" where "therapy" is as defined herein.

[0113] In embodiments, there is provided a crystalline form of Compound (I), for use in providing an inhibitory effect on TEAD.

[0114] In embodiments, there is provided a crystalline form of Compound (I), for use in the treatment of a disease mediated by TEAD, such as cancer.

[0115] In embodiments, there is provided a crystalline form of Compound (I), for use in the treatment of cancer, wherein the cancer is selected from ovarian cancer, cervical cancer, colorectal cancer, breast cancer, pancreatic cancer, glioma, glioblastoma, melanoma, prostate cancer, gastric cancer, lung cancer, hepatocellular cancer (HCC), gastrointestinal stromal tumour (GIST), thyroid cancer, bile duct cancer, endometrial cancer, renal cancer, melanoma and mesothelioma (such as malignant pleural mesothelioma).

[0116] In embodiments, there is provided a crystalline form of Compound (I), for use in the treatment of hippo mutation-positive cancer, such as NF2 mutation-positive or LATS1 / 2 mutation-positive cancer.

[0117] In embodiments, there is provided a crystalline form of Compound (I), for use in the treatment of hippo mutation-positive cancer, such as YAP1 and / or WWTR1 amplified cancer.

[0118] In embodiments, there is provided a crystalline form of Compound (I), for use in the treatment of hippo mutation-positive cancer, such as FAT1 mutant cancer.

[0119] In embodiments, there is provided a crystalline form of Compound (I), for use in the treatment of a cancer driven by YAP or TAZ fusions.

[0120] In embodiments, there is provided a crystalline form of Compound (I), for use in the treatment of a cancer that exhibits an elevated TEAD transcriptional signature. In further embodiments, the cancer that exhibits an elevated TEAD transcriptional signature is hepatocellular cancer (HCC), gastric cancer or prostate cancer. In embodiments, there is provided a crystalline form of Compound (I), for use in the treatment of hippo mutation-positive mesothelioma, such as NF2 mutation-positive or LATS1 / 2 mutation-positive mesothelioma.

[0121] In embodiments, there is provided a crystalline form of Compound (I), for use in the treatment of hippo mutation-positive malignant pleural mesothelioma, such as NF2 mutation-positive or LATS1 / 2 mutation-positive malignant pleural mesothelioma.

[0122] In embodiments, there is provided a crystalline form of Compound (I), for use in the treatment of lung cancer.

[0123] In embodiments, there is provided a crystalline form of Compound (I), for use in the treatment of non-small cell lung cancer.

[0124] In embodiment there is provided crystalline form of Compound (I) for use in the treatment of EGFR mutation-positive cancer (such as non-small cell lung cancer). In further embodiments, the EGFR mutation-positive cancer comprises at least one activating mutation in EGFR selected from exon 19 deletions and L858R substitution mutations. In still further embodiments, the EGFR mutationpositive cancer comprises an EGFR T790M resistance mutation.

[0125] In one aspect of the present specification there is provided the use of a crystalline form of Compound (I), as described herein, in the manufacture of a medicament, such as a medicament for the treatment of cancer.

[0126] In one aspect of the present specification there is provided a method of treating cancer in a patient comprising administering to the patient an effective amount of a crystalline form of Compound (I).

[0127] Combination therapy

[0128] A crystalline form of Compound (I) may also be administered in conjunction with other compounds used for the treatment of the above conditions.

[0129] In embodiments, there is provided a crystalline form of Compound (I), in combination with an additional anti-tumour agent.

[0130] In embodiments, the additional anti-tumour argent is a selected from an EGFR inhibitor, KRAS inhibitor, BRAF inhibitor, CDK4 / 6 inhibitor, MEK inhibitor, MET inhibitor, PI3K inhibitor, AKT inhibitor or ALK inhibitor.

[0131] The additional anti-tumour agent may be a third generation EGFR TKL Third-generation EGFR TKIs are inhibitors of EGFR bearing activating mutations that also significantly inhibit EGFR bearing the T790M mutation and do not significantly inhibit wild-type EGFR. Examples of third-generation TKIs include compounds of Formula (I), osimertinib, AZD3759, lazertinib, nazartinib, CO1686 (rociletinib), HM61713, ASP8273, EGF816, PF-06747775 (mavelertinib), avitinib (abivertinib), alflutinib (AST2818) and CXCK-101 (RX-518), HS-10296 and BPI-7711. Further examples include oritinib (SH-1028), Befotertinib (D-0316), ASK-120067, ZN-e4, YZJ-0318, TL007 XZP (kenaitinib), YK-029A, SLC005-I, TY-9591, XZP-5809-TT1, ZSP0391, and TQB3456.

[0132] In embodiments, there is provided a crystalline form of Compound (I), in combination with osimertinib, or a pharmaceutically acceptable salt thereof.

[0133] In a further aspect there is provided a pharmaceutical composition comprising a crystalline form of Compound (I), as described herein, and a pharmaceutically acceptable excipient.

[0134] Examples

[0135] The specification will now be illustrated by the following non-limiting Examples. Abbreviations used for analytical data, are consistent with the common usage in the field (see J Med Chem Standard Abbreviations and Acronyms http: / / pubsapp.acs.org / paragonplus / submission / jmcmar / jmcmar_abbreviations.pdf). The abbreviation rt means room temperature. For this specification, PERKIN ELMER E-NOTEBOOK was used to name compounds.

[0136] Intermediate 1: 5-fluoro-3-methyl-8-(4-(trifluoromethyl)phenyl)pyrido[4,3-d]pyrimidin-4(3H)-one

[0137] Intermediate 2

[0138] 2-fluoro-4-iodo-N-methylnicotinamide

[0139] Oxalyl chloride (8.20 mL, 93.6 mmol) and DMF (58 pL, 0.75 mmol) were added to a suspension of 2- fluoro-4-iodonicotinic acid (20.0 g, 74.9 mmol) in DCM (150 mL) at 0 °C. The resulting mixture was stirred at 0 °C for 30 min, then rt for 1 hr. The reaction mixture was concentrated to dryness to afford 2-fluoro-4-iodonicotinoyl chloride as an orange solid, which was dissolved in THF (150 mL) and cooled to 0 °C. MeNHz (2 M in THF, 44.9 mL, 89.9 mmol) and triethylamine (13.57 mL, 97.38 mmol) were added, and the resulting mixture was warmed to rt and stirred for 36 hrs. The reaction mixture was diluted with EtOAc (200 mL), and the organic layer was washed with water (80 mL), brine (50 mL), dried over NajSC , filtered, and concentrated to dryness to give the product as a yellow solid. The product was rinsed with hexanes to afford 2-fluoro-4-iodo- / V-methylnicotinamide (19.67 g, 94% yield) as an off-white solid.XH NMR (500 MHz, DMSO-d6) 6 2.79 (3H, d), 7.88 (1H, d), 7.96 (1H, d), 8.57 - 8.69 (1H, m); m / z: (ES+) [M+H]+= 281.

[0140] 4-amino-2-fluoro-N-methylnicotinamide

[0141] Ammonium hydroxide (7.23 mL, 186 mmol) was added to a mixture of 2-fluoro-4-iodo- / V- methylnicotinamide (5.20 g, 18.6 mmol), copper(l) iodide (0.707 g, 3.71 mmol), (2S,4R)-4- hydroxypyrrolidine-2-carboxylic acid (0.974 g, 7.43 mmol), and potassium carbonate (7.70 g, 55.7 mmol) in DMSO (40 mL). The resulting mixture was heated to 50 °C and stirred for 4 hrs. The reaction mixture was cooled to rt, diluted with water (20 mL), and extracted with EtOAc (3 x 50 mL), and the organic layer was washed with brine each time (3 x 15 mL). The combined organics were dried over Na2SO4, filtered, and concentrated to dryness to afford 4-amino-2-fluoro- / V- methylnicotinamide (2.63 g, 84% yield) as an off-white solid.1H NMR (500 MHz, DMSO-d6) 6 2.75 (3H, d), 6.55 (1H, d), 7.08 (2H, br s), 7.63 (1H, d), 8.11 (1H, br s); m / z: (ES+) [M+H]+= 170.

[0142] 4-amino-5-bromo-2-fluoro-N-methylnicotinamide

[0143] Bromine (3.33 mL, 64.7 mmol) was added to a solution of 4-amino-2-fluoro- / V-methylnicotinamide (10.84 g, 64.08 mmol) in AcOH (90 mL) and water (90 mL). The resulting mixture was stirred at rt for 2 hrs. Saturated aq. Na2S20s was added dropwise to the reaction mixture until the orange color disappeared. The reaction mixture was concentrated to dryness, and the crude residue was carefully diluted with saturated aq. NaHCOs (100 mL) and extracted with EtOAc (2 x 200 mL). The combined organics were washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated to dryness to afford 4-amino-5-bromo-2-fluoro- / V-methylnicotinamide (12.4 g, 78% yield) as a faint yellow / off- white solid.XH NMR (500 MHz, DMSO-d6) 6 2.76 (3H, d), 7.08 (2H, br s), 8.02 (1H, s), 8.39 (1H, br s); m / z: (ES+) [M+H]+= 248.

[0144] Intermediate 1: 8-bromo-5-fluoro-3-methylpyrido[4,3-d]pyrimidin-4(3H)-one

[0145] A mixture of 4-amino-5-bromo-2-fluoro- / V-methylnicotinamide (1.02 g, 4.11 mmol), triethoxymethane (18.00 mL, 108.2 mmol), and 4-methylbenzenesulfonic acid hydrate (0.782 g, 4.11 mmol) in NMP (4.0 mL) was heated to 75 °C and stirred for 22 hrs. The reaction mixture was cooled to rt. The precipitate was collected by filtration and washed with water and EtOAc. The filtrate was extracted with EtOAc. The combined organics were dried over Na2SO4, filtered, and concentrated to dryness. The crude material was diluted with MeOH (20 mL) and water (40 mL) and sonicated for a few minutes. The precipitate was collected by filtration and washed with 50% EtOAc / hexanes and dried to afford 8-bromo-5-fluoro-3-methylpyrido[4,3-d]pyrimidin-4(3H)-one (Intermediate 1, 0.899 g, 85% yield) as a white solid.XH NMR (500 MHz, DMSO-d6) 6 3.48 (s, 3H), 8.68 (s, 1H), 8.74 (s, 1H); m / z: (ES+) [M+H]+= 258.

[0146] Intermediate 25-fluoro-3-methyl-8-(4-(trifluoromethyl)phenyl)pyrido[4,3-d]pyrimidin-4(3H)-one

[0147] A mixture of 8-bromo-5-fluoro-3-methylpyrido[4,3-d]pyrimidin-4(3H)-one (Intermediate 2, 3.369 g, 13.06 mmol), (4-(trifluoromethyl)phenyl)boronic acid (3.220 g, 16.97 mmol), and potassium carbonate (3.610 g, 26.11 mmol) in 1,4-dioxane (50 mL) and water (5 mL) was degassed and purged with nitrogen (3x). PdCb(dppf) (0.096 g, 0.13 mmol) was added, and the reaction mixture was degassed and purged with nitrogen (3x). The reaction was heated to 55 °C and stirred for 16 hrs under N2. The reaction mixture was cooled to rt, diluted with H2O (150 mL), stirred at rt for 30 minutes, and filtered. The solid was collected and washed with H2O (100 mL), hexanes (100 mL), and dried under high vacuum to give a crude solid. The solid was dissolved in EtOAc (50 mL), filtered through a pad of diatomaceous earth, and washed with EtOAc (100 mL). The filtrate was concentrated to dryness and then diluted with 1:6 TBME / hexane (35 mL), sonicated, and filtered to give product, which was diluted again with 1:6 TBME / hexane (35 mL), stirred at rt for 2 hrs, and filtered to afford 5-fluoro-3-methyl-8-(4-(trifluoromethyl)phenyl)pyrido[4,3-d]pyrimidin-4(3H)-one (Intermediate 2, 3.324 g, 79% yield) as a tan solid. More trituration with 1:4 TBME / hexanes could be done as needed depending on purity.1H NMR (500 MHz, DMSO-d6) 6 3.49 (3H, s), 7.80 - 7.87 (4H, m), 8.52 (1H, s), 8.62 (1H, s); m / z: (ES+) [M+H]+= 324.

[0148] Compound (I): 5-(((3S,5 / ?)-5-hydroxytetrahydro-2H-pyran-3-yl)amino)-3-methyl-8-(4- (trifluoromethyl)phenyl)pyrido[4,3-c / ]pyrimidin-4(3H)-one A mixture of 5-fluoro-3-methyl-8-(4-(trifluoromethyl)phenyl)pyrido[4,3-c / ]pyrimidin-4(3H)-one (Intermediate 2, 50.0 mg, 0.155 mmol), (3R,5S)-5-aminotetrahydro-2H-pyran-3-ol hydrochloride (28.5 mg, 0.186 mmol), and DIPEA (0.081 mL, 0.46 mmol) in DMSO (0.5 mL) was heated to 70 °C and stirred for 1 hr. The reaction mixture was cooled to rt and directly purified by silica flash chromatography (0 to 100% EtOAc in hexanes) to afford 5-(((3S,5R)-5-hydroxytetrahydro-2H-pyran- 3-yl)amino)-3-methyl-8-(4-(trifluoromethyl)phenyl)pyrido[4,3-d]pyrimidin-4(3H)-one (Compound (I) Form A, 60.9 mg, 94% yield) as a white solid.XH NMR (500 MHz, DMSO-d6) 6 1.54 (1H, dt), 2.22 - 2.30 (1H, m), 3.14 - 3.23 (2H, m), 3.47 (3H, s), 3.68 (1H, tq), 3.74 (1H, dd), 3.90 (1H, dd), 4.30 (1H, qt), 5.03 (1H, d), 7.76 (4H, s), 8.34 (1H, s), 8.46 (1H, s), 9.20 (1H, br d); m / z: (ES+) [M+H]+= 421.

[0149] Example 1: Compound (I) Form A

[0150] Compound (I) Form A was analyzed by XRPD, as shown in Figure 1. Compound (I) Form A was analyzed by thermal techniques. DSC analysis shows an endothermic event of de-solvation with an onset at 29 °C and a peak of 48 °C and an endothermic event of melting with an onset at 120 °C and a peak of 127 °C. A representative DSC thermogram of Form A is shown in Figure 2. TGA indicated that Form A exhibits a mass loss of about 3.8 % upon heating from about 25 °C to about 100 °C. A representative TGA thermogram of Form A is shown in Figure 3.

[0151] Example 2: Compound (I) Form B

[0152] 10 mg of Compound (I) Form A was suspended in 0.20 mL of water. The suspension was stirred at the room temperature for 1 day. Crystalline solid of Compound (I) Form B was obtained after filtration and drying.

[0153] Compound (I) Form B was analyzed by XRPD as shown in Figure 4. Compound (I) Form B was analyzed by thermal techniques. DSC analysis shows an endothermic event of de-solvation with an onset at 29 °C and a peak of 57 °C and an endothermic event of melting with an onset at 114 °C and a peak of 123 °C. A representative DSC thermogram of Form B is shown in Figure 5. TGA indicated that Form B exhibits a mass loss of about 6.2 % upon heating from about 25 °C to about 100 °C. A representative TGA thermogram of Form B is shown in Figure 6.

[0154] Example 3: Compound (I) Form C

[0155] 20 mg of Compound (I) Form A was suspended in 0.40 mL of 1% polysorbate 80 aqueous solution. The suspension was stirred at the room temperature for 1 day. 15 mg of crystalline solid of Compound (I) Form C was obtained after filtered and dried. Compound (I) Form C was analyzed by XRPD , as shown in Figure 7. Compound (I) Form C was analyzed by thermal techniques. DSC analysis shows an endothermic event of de-solvation with an onset at 63 °C and a peak of 92 °C. A representative DSC thermogram of Form C is shown in Figure 8. TGA indicated that Form C exhibits a mass loss of about 3.6 % upon heating from about 25 °C to about 100 °C. A representative TGA thermogram of Form C is shown in Figure 9.

[0156] Biological Data

[0157] The data in Table 1 were generated using the Compound (I) of the present specification and the assays described below.

[0158] (i) TEAD4 FRET

[0159] Compounds were dosed with a final DMSO concentration of 1% (v / v). Compound IC5o values were assessed following a 10-point, half-logio dilution schema starting at 100 pM compound concentration. Specifically, human TEAD protein from TEAD4(217-434) was cloned into an overexpression vector, expressed as an / V-terminal HIS-TEV-Avi-tagged fusion protein in E coli, and subsequently purified, then protein was chemically depalmitoylated & biotinylated. The assay was performed in 384-well LV plates (384-well black, medium binding, PS, HIBASE, GREINER #784076) and run in the presence and absence of the compound of interest. Each well of 5 pL assay mixture contained 10 mM Tris-HCI (pH 7.5), 100 mM NaCI, 0.05 mM EDTA, 1 mM TECP, 1% DMSO, 0.03% Pluronic acid F127, 20 nM dePai Avi TEAD4 (217-434) -depalmitoylated & biotinylated protein, 0.8 nM Streptavidin Terbium cryptate (CisBio#610SATLB), 625 nM FAM labelled Probe A. Reactions were incubated at 25°C for 120 min before reading on a PHERASTAR FSX Plate Reader (337 520 490 HTRF module required) (Supplier BMP).

[0160] The data file from the PHERAstar FSX contains both the acceptor (520 nm) and donor channels (490 nm) ("Channel A" = acceptor channel (520 nm), "Channel B" = donor channel (490 nm)). The ratio of the donor and acceptor (Channel A / Channel B) is calculated within Genedata Assay Analyzer. Subsequently, the dose-response of the ratio to testing compound concentration was fitted to a select fit model that will provide the best fit quality using automatic parameter (SMARTFIT) to derive IC5o values for each testing compound.

[0161] Probe A is 3',6'-dihydroxy-3-oxo-N-{8-[2-(5-{2-[4-(trifluoromethyl)anilino]phenyl}-2H-tetrazol-2- yl)acetamido]octyl}-3H-spiro[[2]benzofuran-l,9'-xanthene]-5-carboxamide;

[0162] (ii) TEAD Reporter Assay

[0163] MCF7-Tead cell line was obtained from BPS BIOSCIENCE (catalog number 60618) and was maintained in DMEM containing 10% fetal calf serum, 2mM glutamine, and 400pg / ml G418. Cells were grown in a humidified incubator at 37 °C with 5% CO2. Cells were distributed to flat bottom white polystyrene TC treated 384 well plates at a density of 3,500 cells / well in 30pL. Cells were incubated for 24 hours at 37 °C with 5% CO2. Cells were acoustically dosed using an Echo 555, with compounds serially diluted in 100% DMSO. Plates were incubated for an additional 24 hours. Cells were examined for luciferase activity through the addition of 30pL Bright-Glo luciferase (Promega catalog number E2620). Plates were incubated for 10 minutes at room temperature and luminescence was read using Tecan plate reader. The data obtained with each compound was exported to GENEDATA software to perform curve fitting analysis. IC50 value was calculated based on the concentration of compound that is required to give a 50% effect compared to DMSO control.

[0164] (iii) Control reporter (MCF7-Luciferase)

[0165] MCF7-Luciferase cell line was obtained from GENTARGET (catalog number SC050-L) and was maintained in DMEM containing 10% fetal calf serum, 2mM glutamine. Cells were grown in a humidified incubator at 37°C with 5% CO2. Cells were distributed to flat bottom white polystyrene TC treated 384 well plates at a density of 3,500 cells / well in 30pL. Cells were incubated for 24 hours at 37 °C with 5% CO2. Cells were acoustically dosed using an Echo 555, with compounds serially diluted in 100% DMSO. Plates were incubated for an additional 24 hours. Cells were examined for luciferase activity through the addition of 30uL Bright-Glo luciferase (Promega catalog number E2620). Plates were incubated for 10 minutes at room temperature and luminescence was read using Tecan plate reader. The data obtained with each compound was exported to GENEDATA software to perform curve fitting analysis. IC50 value was calculated based on the concentration of compound that is required to give a 50% effect compared to DMSO control.

[0166] Table 1

[0167] Synthesis of Probe A tert-butyl (8-(2-(5-(2-((4-(trifluoromethyl)phenyl)amino)phenyl)-2H-tetrazol-2- yl)acetamido)octyl)carbamate l-[Bis(dimethylamino)methylene]-lH-l,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate (HATU, 110 mg, 0.29 mmol) and DIPEA (84 pL, 0.48 mmol) were added to a solution of 2-(5-(2-((4- (trifluoromethyl)phenyl)amino)phenyl)-2H-tetrazol-2-yl)acetic acid (70 mg, 0.19 mmol, disclosed in WO2018204532, the contents of which are incorporated by reference) and tert-butyl (8- aminooctyl)carbamate (71 mg, 0.29 mmol) in DMF (1.8 mL) at 0 °C, and the reaction mixture was stirred for 3 hrs while slowly warming to rt. The crude reaction mixture was diluted with EtOAc (20 mL) and washed with saturated aq. NaHCOs (2 x 20 mL), saturated aq. NH4CI (2 x 20 mL) and brine (20 mL). The organic layer was dried over NajSC , filtered and concentrated to dryness. The crude material was purified by flash silica chromatography (0-50% EtOAc in Hex) to afford tert-butyl (8-(2- (5-(2-((4-(trifluoromethyl)phenyl)amino)phenyl)-2H-tetrazol-2-yl)acetamido)octyl)carbamate (53.7 mg, 47% yield) as a white solid.XH NMR (500 MHz, DMSO-d6) 6 1.22 (9H, br s), 1.29 - 1.38 (9H, m), 1.41 (3H, br d), 2.87 (2H, q), 3.09 (2H, q), 5.49 (2H, s), 6.72 (1H, br t), 7.13 - 7.19 (1H, m), 7.24 (2H, br d), 7.45 - 7.50 (1H, m), 7.52 - 7.59 (3H, m), 8.03 (1H, dd), 8.40 (1H, br t), 8.77 (1H, s); m / z: (ES+) [M+H]+= 590.

[0168] N-(8-aminooctyl)-2-(5-(2-((4-(trifluoromethyl)phenyl)amino)phenyl)-2H-tetrazol-2-yl)acetamide

[0169] TFA (0.50 mL, 6.5 mmol) was added to a solution of tert-butyl (8-(2-(5-(2-((4- (trifluoromethyl)phenyl)amino)phenyl)-2H-tetrazol-2-yl)acetamido)octyl)carbamate (50 mg, 0.08 mmol) in DCM (1 mL) at 0 °C and the reaction mixture was stirred for 45 min. The reaction mixture was concentrated to dryness to afford / V-(8-aminooctyl)-2-(5-(2-((4- (trifluoromethyl)phenyl)amino)phenyl)-2H-tetrazol-2-yl)acetamide TFA as a pale blue oil. 3',6'-dihydroxy-3-oxo-N-(8-(2-(5-(2-((4-(trifluoromethyl)phenyl)amino)phenyl)-2H-tetrazol-2- yl)acetamido)octyl)-3H-spiro[isobenzofuran-l,9'-xanthene]-5-carboxamide (Probe A)

[0170] HATU (61 mg, 0.16 mmol) and DIPEA (86 pL, 0.49 mmol) were added to a solution of 5- carboxyfluorescein (60 mg, 0.16 mmol) and / V-(8-aminooctyl)-2-(5-(2-((4- (trifluoromethyl)phenyl)amino)phenyl)-2H-tetrazol-2-yl)acetamide (60 mg, 0.12 mmol) in DMF (1.1 mL) at 0 °C. The reaction mixture was stirred at 0 °C for 2 hrs before warming to rt with stirring for an additional 1 h. The reaction mixture was diluted with EtOAc (50 mL) and washed with saturated aq. NH4 (2 x 30 mL) and brine (30 mL). The organic layer was dried over NajSC , filtered and concentrated to dryness. The resulting residue was purified by preparative HPLC (Column = Xbridge C18, 4.6 mm x 50 mm, 5 pm; Gradient = 13 to 95% MeCN in H2O w / 0.2% NH4OH over 4 min; Flow rate = 0.6 mL / min; UV detection @ 254) to afford 3',6'-dihydroxy-3-oxo- / V-(8-(2-(5-(2-((4- (trifluoromethyl)phenyl)amino)phenyl)-2H-tetrazol-2-yl)acetamido)octyl)-3H-spiro[isobenzofuran- l,9'-xanthene]-5-carboxamide (82 mg, 79 %) as a bright red solid.1H NMR (500 MHz, DMSO-d6) 6 1.29 (9H, br d), 1.39 - 1.48 (2H, m), 1.49 - 1.59 (2H, m), 2.53 (2H, br s), 3.10 (2H, q), 5.50 (2H, s), 6.45 (2H, br d), 6.54 (2H, br s), 6.58 (2H, s), 7.12 - 7.19 (1H, m), 7.24 (2H, d), 7.28 (1H, d), 7.43 - 7.50 (1H, m), 7.54 (3H, t), 8.03 (1H, dd), 8.11 (1H, br d), 8.43 (1H, s), 8.50 (1H, br t), 8.72 (1H, br t), 8.78 (1H, s); m / z: (ES+) [M+H]+= 848.

[0171] The above description of illustrative embodiments is intended only to acquaint others skilled in the art with the Applicant's specification, its principles, and its practical application so that others skilled in the art may readily adapt and apply the specification in its numerous forms, as they may be best suited to the requirements of a particular use. This description and its specific examples, while indicating embodiments of this specification, are intended for purposes of illustration only. This specification, therefore, is not limited to the illustrative embodiments described in this specification, and may be variously modified. In addition, it is to be appreciated that various features of the specification that are, for clarity reasons, described in the context of separate embodiments, also may be combined to form a single embodiment. Conversely, various features of the specification that are, for brevity reasons, described in the context of a single embodiment, also may be combined to form sub-combinations thereof.

Claims

Claims1. A crystalline form of 5-(((3S,5R)-5-hydroxytetrahydro-2H-pyran-3-yl)amino)-3-methyl-8-(4- (trifluoromethyl)phenyl)pyrido[4,3-d]pyrimidin-4(3H)-one (Form C), which has an X-ray powder diffraction pattern with specific peaks at 2-theta = 6.4 and 9.7° when measured using CuKa radiation, wherein said values may be plus or minus 0.2° 2-theta.

2. A crystalline form as claimed in claim 1, which has an X-ray powder diffraction pattern with specific peaks at 2-theta = 6.4, 9.7, 16.9, 20.3, and 21.0° when measured using CuKa radiation, wherein said values may be plus or minus 0.2° 2-theta.

3. A crystalline form as claimed in claim 1 or claim 2, which has an X-ray powder diffraction pattern with specific peaks at 2-theta = 6.4, 9.7, 13.3, 16.1, 16.9, 19.3, 20.3, 21.0, 22.2, and 24.1° when measured using CuKa radiation, wherein said values may be plus or minus 0.2° 2-theta.

4. A crystalline form of 5-(((3S,5R)-5-hydroxytetrahydro-2H-pyran-3-yl)amino)-3-methyl-8-(4- (trifluoromethyl)phenyl)pyrido[4,3-d]pyrimidin-4(3H)-one (Form C), which has an X-ray powder diffraction pattern substantially as shown in Figure 7, when measured using CuKa radiation.

5. A crystalline form of 5-(((3S,5R)-5-hydroxytetrahydro-2H-pyran-3-yl)amino)-3-methyl-8-(4- (trifluoromethyl)phenyl)pyrido[4,3-d]pyrimidin-4(3H)-one (Form B), which has an X-ray powder diffraction pattern with specific peaks at 2-theta = 5.4° and 10.9° when measured using CuKa radiation, wherein said values may be plus or minus 0.2° 2-theta.

6. A crystalline form as claimed in claim 5, which has an X-ray powder diffraction pattern with specific peaks at 2-theta = 5.4, 10.9, 14.2, 19.8, and 23.7° when measured using CuKa radiation, wherein said values may be plus or minus 0.2° 2-theta.

7. A crystalline form as claimed in claim 5 or claim 6, which has an X-ray powder diffraction pattern with specific peaks at about 2-theta = 5.4, 10.9, 13.4, 14.2, 18.9, 19.3, 19.8, 22.4, 22.9, and 23.7° when measured using CuKa radiation, wherein said values may be plus or minus 0.2° 2-theta.

8. A crystalline form of 5-(((3S,5R)-5-hydroxytetrahydro-2H-pyran-3-yl)amino)-3-methyl-8-(4-(trifluoromethyl)phenyl)pyrido[4,3-d]pyrimidin-4(3H)-one (Form B), which has an X-ray powder diffraction pattern substantially as shown in Figure 4, when measured using CuKa radiation.

9. A pharmaceutical composition comprising a crystalline form as claimed in any one of claims 1 to 8, and a pharmaceutically acceptable excipient.

10. A crystalline form as claimed in any one of claims 1 to 8, for use in therapy.

11. A crystalline form as claimed in any one of claims 1 to 8, for use in the treatment of cancer.

12. A method of treating cancer in a patient comprising administering to the patient a crystalline form, as claimed in any one of claims 1 to 8.

Images