Drug compounds

Abstract

The present invention provides a composition of matter, which: (i) consists of at least 90% by weight of an atropisomer (2A) and 0-10% by weight of an atropisomer of formula (2B); or (ii) consists of at least 90% by weight of an atropisomer (2B) and 0-10% by weight of an atropisomer of formula (2A); wherein the atropisomer of formula (2A) and the atropisomer of formula (2B) are represented by the following: Formula (2A) and Formula (2B), or pharmaceutically acceptable salts or tautomers thereof, wherein ring X is a benzene or pyridine ring; ring Y is selected from a benzene ring, a pyridine ring and a thiophene ring; R 1 is trifluoromethyl; R 2 is hydrogen; R 3 is hydrogen; m is 0 or 1; n is 0, 1 or 2; Ar 1 is a monocyclic aromatic ring selected from the group consisting of benzene and pyridine; each monocyclic aromatic ring is unsubstituted or substituted with 1 or 2 substituents R as defined herein 5 Replacement; and R 4 ; When present, R 5 , R 6 and R 7 Independently selected from various substituents as defined herein. Also provided are individual atropisomers thereof and individual atropisomers of various compounds having a five-membered heteroaromatic ring, wherein the heteroaromatic ring contains 1 or 2 nitrogen atoms or 1 nitrogen and 1 oxygen atom, wherein the three rings Ar1, X and Y, and the substituents R1-R7 are as defined in the following formula (1A) or (1B); pharmaceutical compositions, and uses of the atropisomers and compositions, which are inhibitors of PLK1- and PLK4 kinases, for example, for the treatment of cancer. Formula (1A), Formula (1B).

Description

[0001] The present invention relates to atropisomers of triarylpyrrole derivatives and analogs thereof, methods for preparing the same, pharmaceutical compositions containing the same and their use in treating diseases such as cancer. Background Art

[0002] The protein expressed from the normal KRAS gene plays an important role in normal tissue signaling. Mutations in the KRAS gene due to single amino acid substitutions, and especially single nucleotide substitutions, are the cause of activating mutations, which are necessary steps in the development of many cancers. The resulting mutant proteins are involved in various malignancies, including lung adenocarcinoma, mucinous adenoma, pancreatic ductal carcinoma, and colorectal cancer. Similar to other members of the Ras family, the KRAS protein is a GTPase and is involved in many signal transduction pathways.

[0003] KRAS acts as a molecular on / off switch. Once turned on, it recruits and activates proteins necessary for the propagation of growth factors and other receptor signals, such as c-Raf and PI-3 kinase. Normal KRAS binds to GTP in the active state and has an intrinsic enzymatic activity, namely cleaving the terminal phosphate of the nucleotide and converting it to GDP. After converting GTP to GDP, KRAS is turned off. The conversion rate is usually slow, but can be significantly accelerated by auxiliary proteins such as RasGAP of the GTPase activating protein (GAP) class. In turn, KRAS can bind to proteins of the guanine nucleotide exchange factor (GEF) class, such as SOS1, which forces the release of the bound nucleotide. Subsequently, KRAS binds to GTP present in the cytosol, and GEF is released from ras-GTP. In mutant KRAS, its GTPase activity is directly removed, so that KRAS is constitutively active. Mutant KRAS is usually characterized by mutations in codons 12, 13, 61 or a mixture thereof.

[0004] It is known that the viability of cancer cells carrying mutant KRAS depends on Polo-like kinase 1 (PLK1), and silencing PLK1 has been shown to cause cell death containing mutant KRAS (see Luo et al., Cell. 2009 May 29; 137(5): 835–848). Therefore, compounds that inhibit PLK1 can be used to treat cancers caused by KRAS mutations, but compared with other kinases, kinase inhibitors currently designed to bind to the conserved ATP binding domain of PLK1 may not be able to enter this mode of action due to lack of selectivity (see, for example, Elsayed et al., Future Med. Chem. (2019) 11(12), 1383–1386).

[0005] PLK1 is a serine / threonine kinase consisting of 603 amino acids and having a molecular weight of 66 kDa, and is an important regulator of the cell cycle. In particular, PLK1 is important for mitosis and is involved in the formation and changes of the mitotic spindle during the M phase of the cell cycle and the activation of the CDK / cyclin complex.

[0006] All Polo-like kinases contain an N-terminal serine / threonine kinase catalytic domain and a C-terminal region containing one or two Polo boxes (Lowery et al., Oncogene, (2005), 24, 248-259). For Polo-like kinases 1, 2, and 3, the entire C-terminal region (including two Polo boxes) acts as a single modular phosphoserine / threonine binding domain, called the Polo box domain (PBD). In the absence of a bound substrate, the PBD inhibits the basal activity of the kinase domain. The phosphorylation-dependent binding of the PBD to its ligand releases the kinase domain, while simultaneously localizing the Polo-like kinase to a specific subcellular structure.

[0007] It has been shown (Reindl et al., Chemistry & Biology, 15, 459-466, May 2008) that since PLKL1 is localized to its intracellular anchoring site via its polo box domain, the action of PLK1 can be inhibited by small molecules that interfere with its intracellular localization by inhibiting the function of the PBD.

[0008] The tumor protein p53 functions as a tumor suppressor and plays a role in apoptosis, genomic stability, and angiogenesis inhibition. It is known that tumors with both p53 deficiency and high PLK1 expression may be particularly sensitive to PLK1 inhibitors (Yim et al., Mutat Res Rev Mutat Res, (2014). 761, 31-39).

[0009] Therefore, evidence from the literature suggests that small molecules that bind to and inhibit PBD function should be effective inhibitors of PLK1 kinase and, therefore, also useful for treating cancers caused by KRAS and / or p53 mutations. In particular, since the PBD domain is only present in PLK, inhibitors designed against this domain have the potential to be more selective than previous ATP-competitive inhibitors, enabling a greater ability to target KRAS mutant and p53-deficient cancers.

[0010] The identification and development of drugs for the treatment of primary brain cancer has proven to be particularly challenging. Targeted cancer therapies, and in particular therapies using protein kinase inhibitors, have become a major focus for pharmaceutical and biotechnology companies (Nature Reviews Clinical Oncology 2016, 13, 209--227). However, despite the approval of more than thirty kinase inhibitors for oncology, none of these kinase inhibitors have been used to treat primary brain cancer. A particular problem is that most approved kinase inhibitor oncology drugs lack the necessary drug qualities required to achieve brain exposure if they are to be used to treat brain cancer [JMC 2016, 59(22), 10030-10066].

[0011] Alkylating agent temozolomide Temozolomide is currently the first-line treatment for the brain cancer glioblastoma multiforme, and is often combined with radiation therapy. However, drug resistance is a major problem in the management of glioblastoma and therefore limits the effectiveness of temozolomide. Therefore, currently, malignant glioblastoma remains incurable.

[0012] Polo-like kinase 1 (PLK1) is overexpressed in multiple tumor types, including glioblastoma multiforme (Translational Oncology 2017, 10, 22-32). In addition, recent studies have shown that PLK1 drives checkpoint adaptation and resistance to temozolomide in glioblastoma multiforme [Oncotarget 2017, 8, 15827-15837].

[0013] Ependymomas are tumors of the brain and spinal cord where the current standard of care is limited to surgery and radiation. PLK1 has been implicated in ependymomas, and inhibitors of PLK1 are active against ependymoma cell lines [Gilbertson et. al., Cancer Cell (2011) 20, 384-399].

[0014] PLK1 has also been studied as a target for Diffuse Intrinsic Pontine Glioma (DIPG), a high grade, aggressive childhood brain tumor [Amani et al. BMC Cancer (2016) 16, 647 and Cancer Biology and Therapy (2018) 19, 12, 1078-1087].

[0015] More specifically, inhibition of PLK1 has been shown to enhance the efficacy of temozolomide against IDH1 mutant gliomas [Oncotarget, (2017) 8, 9, 15827-15837] and to inhibit tumor growth in an MMR-deficient, temozolomide-resistant glioblastoma xenograft model [Mol Cancer Ther; 17(12) December 2018].

[0016] In the above cases, current inhibitors lack adequate brain exposure.

[0017] Compounds that inhibit PLKL1 but do not induce drug resistance and exhibit good brain exposure are expected to be useful in the treatment of glioblastoma multiforme and other brain cancers.

[0018] PLK4 is a member of the polo-like kinase family of serine / threonine kinases that plays a key role in centrosome duplication, acting as a central regulator of centriole duplication (Bettencourt-Dias, Curr Biol. 2005 15(24); 2199-207). PLK4-dependent changes in centrosomes can lead to asymmetric chromosome segregation during mitosis, which can trigger cell death after chromosome missegregation and mitotic defects.

[0019] PLK4 is aberrantly expressed in human cancers and is involved in tumorigenesis and metastasis. Therefore, PLK4 has been highlighted as a promising target for cancer therapy (Zhao, J Canc Res Clin Oncol., 2019).

[0020] PLK4 is overexpressed in many cancers, including rhabdoid tumors, medulloblastomas and other embryonal brain tumors (Pediatr Blood Cancer. 2017), as well as breast, lung, melanoma, gastric, colorectal, pancreatic and ovarian cancers. Elevated or overactivated PLK4 is associated with poor survival in patients with cancer, including ovarian, breast and lung cancer (Zhao, J Canc Res Clin Oncol., 2019).

[0021] PLK4 inhibition has been studied for the treatment of glioblastoma multiforme, and PLK4 has been shown to play a key role in regulating temozolomide chemosensitivity. The combination of temozolomide and PLK4 inhibition in glioblastoma PDX models has been shown to enhance anti-tumor effects compared to temozolomide alone (Cancer Letters, Vol 443, 2019, 91-107).

[0022] PLK4 is reported to cooperate with p53 inactivation in cancer development, and cancers with PLK4 overexpression and p53 deficiency are predicted to be prone to tumor formation (Sercin, 2016; Nat Cell Biol 18: 100-110). Therefore, compounds that inhibit PLK4 activity would be expected to be useful in the treatment of p53 mutant cancers.

[0023] Inhibition of PLK4 leads to anti-tumor activity in lung cancer, with activity seen in cancers carrying wild-type and mutant KRAS (Kawakami, PNAS 2018, 115(8)1913-18). Therefore, compounds that inhibit PLK4 activity would be expected to be useful in the treatment of KRAS mutant cancers.

[0024] Current PLK4 inhibitors act at the kinase active site and are not optimal for brain penetration (Int. J. Mol. Sci. 2019, 20, 2112). Therefore, compounds that inhibit PLK4 PBD but also show good brain exposure are expected to be used to treat glioblastoma multiforme and other brain cancers.

[0025] Our earlier international patent application WO2018 / 197714 discloses a compound of formula (0):

[0026]

[0027] Wherein ring X is a benzene or pyridine ring, ring Y is a benzene, pyridine, thiophene or furan ring, Ar 1 is an optionally substituted benzene, pyridine, thiophene or furan ring, and R 1 To R 4 , R 6 , R 7 is hydrogen or various substituents. These compounds are described as having anticancer activity and good brain exposure after oral administration, making them good candidates for the treatment of brain cancer. These compounds are active against glioblastoma cell lines and are believed to act as inhibitors of the Polo box domain of PLK1 kinase. These compounds are also disclosed to be active against mutant-RAS cancer cell lines (such as HCT116) and can also be used to treat cancers caused by KRAS mutations. Summary of the invention

[0028] It has now been found that compounds of the type disclosed in our earlier application form atropisomers in which R 1is a methyl group or a substituent of larger size, and in particular a trifluoromethyl group. Atropisomers are stereoisomers caused by hindered rotation about a single bond axis, where the energy barrier to the rotation barrier is high enough to allow separation of individual rotamers; see LaPlante et al., J. Med. Chem., 54:7005-7022 (2011).

[0029] Atropisomers can be divided into three classes based on the amount of energy required to racemize by rotation about the chiral axis and the length of time required for racemization to occur. Class 1 atropisomers have a rotation barrier about the chiral axis of <84 kJ / mol (20 kcal / mol) and racemize within time periods measured in minutes or less at room temperature; Class 2 atropisomers have a rotation barrier between 84 and 117 kJ / mol (20–28 kcal / mol) and racemize within time periods measured in hours to months at room temperature; and Class 3 atropisomers have a rotation barrier >117 kJ / mol (28 kcal / mol) and racemize within time periods measured in years at room temperature.

[0030] Can be used Figure 1 The Cahn-Ingold-Prelog R versus S system shown in Figure 1 classifies atropisomers and is illustrated by (S)-6,6'-dinitrobiphenyl-2,2'-dicarboxylic acid.

[0031] In this system, the nearest substituent on either side of the aryl-aryl bond is given priority in the order abcd. Since the substituents a, b, and c are in a counterclockwise arrangement, the atropisomer is the S isomer. In the corresponding R isomer, the substituents a, b, and c are in a clockwise arrangement.

[0032] The atropisomer compounds of the present invention are sufficiently stable to be separated and characterized and have been found not to undergo racemization to any significant extent, even when heated to temperatures up to 80°C for a period of 10 days. Thus, the atropisomers of the present invention can be classified as Class 3 atropisomers. It is believed that atropisomerism occurs because the substituent R 1 With aromatic ring Ar 1 The steric interaction between Z and Y prevents rotation about the bond between ring Z and X.

[0033] Individual atropisomers of a given pair have been found to have significantly different biological properties. Thus, typically, one atropisomer of a pair has significantly more activity against certain cancer targets than the other atropisomer of the pair.

[0034] According to a first embodiment (embodiment 1.1), the present invention provides:

[0035] (i) a composition of matter consisting of at least 90% by weight of the atropisomer (1A) and 0-10% by weight of the atropisomer of formula (1B); or

[0036] (ii) a composition of matter consisting of at least 90% by weight of an atropisomer (1B) and 0-10% by weight of an atropisomer of formula (1A);

[0037] The atropisomer of formula (1A) and the atropisomer of formula (1B) are represented by the following:

[0038]

[0039] or a pharmaceutically acceptable salt or tautomer thereof, wherein

[0040] Z is a 5-membered heteroaryl ring comprising one or two nitrogen ring members and optionally one further heteroatom ring member selected from N and O;

[0041] Ring X is a 6-membered carbocyclic or heterocyclic aromatic ring containing 0, 1 or 2 nitrogen heteroatom ring members;

[0042] Ring Y is a 6-membered carbocyclic ring or a 5-membered or 6-membered heterocyclic aromatic ring containing 1 or 2 heteroatom ring members selected from N, O and S;

[0043] Ar 1 is a monocyclic 5- or 6-membered aromatic ring, optionally containing 0, 1 or 2 heteroatom ring members selected from N, O and S and optionally substituted by one or more substituents R 5 replace;

[0044] m is 0, 1 or 2;

[0045] n is 0, 1 or 2;

[0046] R 1 Selected from:

[0047] -chlorine;

[0048] -bromine;

[0049] -hydroxyl group;

[0050] - cyano;

[0051] -carboxyl;

[0052] -C(O)O(Hyd 1 );

[0053] -CONH2;

[0054] -amino group;

[0055] --(Hyd 2 )NH;

[0056] -(Hyd 2 )2N; and

[0057] -C 1-5 A hydrocarbon group, wherein 0, 1 or 2 carbons in the hydrocarbon group are replaced by heteroatoms selected from N, O and S, and the hydrocarbon group is optionally substituted by one or more fluorine atoms;

[0058] Hyd 1 、Hyd 1a 、Hyd 1b 、Hyd 2 、Hyd 2a 、Hyd 2b and Hyd 2c Same or different and C 1-4 hydrocarbon groups;

[0059] R 2 Selected from hydrogen and C 1-4 hydrocarbon groups;

[0060] R 3 Selected from hydrogen and C 1-4 hydrocarbon groups;

[0061] R 4 Selected from:

[0062] -fluorine;

[0063] -chlorine;

[0064] -bromine;

[0065] -Hydroxyl;

[0066] - cyano;

[0067] -carboxyl;

[0068] -C(O)O(Hyd 1a );

[0069] -CONH2;

[0070] -amino group;

[0071] --(Hyd 2a )NH;

[0072] -(Hyd 2a )2N; and

[0073] -C 1-5A hydrocarbon group, wherein 0, 1 or 2 carbons in the hydrocarbon group are replaced by heteroatoms selected from N, O and S, and the hydrocarbon group is optionally substituted by one or more fluorine atoms;

[0074] R 5 Selected from halogen; O-Ar 2 ; Cyano, Hydroxyl; Amino; Hyd 1b -SO2- and non-aromatic C 1-8 A hydrocarbon group, wherein 0, 1 or 2 carbons, but not all carbons, in the hydrocarbon group are optionally replaced by heteroatoms selected from N, O and S, and wherein the hydrocarbon group is optionally substituted with one or more fluorine atoms;

[0075] Ar 2 is a phenyl, pyridyl or pyridone group, optionally substituted by 1 or 2 substituents selected from halogen; cyano and C optionally substituted by one or more fluorine atoms; 1-4 hydrocarbon groups;

[0076] R 6 is selected from halogen, cyano, nitro and the group Q 1 -R a -R b ;

[0077] Q 1 Does not exist or C 1-6 Saturated hydrocarbon joints;

[0078] R a Not present or selected from O; C(O); C(O)O; CONR c ; N(R c )CO; N(R c )CONR c ,NRc;S;SO;SO2;SO2NR c ; and NR c SO2;

[0079] R b Selected from:

[0080] -hydrogen;

[0081] -C 1-8 A non-aromatic hydrocarbon group, wherein 0, 1 or 2 carbon atoms in the hydrocarbon group are replaced by heteroatoms selected from N and O, and the C 1-8 The non-aromatic hydrocarbon group is optionally substituted by one or more selected from fluorine and the group Cyc 1 substituted with a substituent; and

[0082] -Cyc Group 1 ;

[0083] Cyc 1is a non-aromatic 4-7 membered carbocyclic or heterocyclic group containing 0, 1 or 2 heteroatom ring members selected from N, O and S, and optionally substituted by one or more selected from hydroxyl; amino; (Hyd 2c )NH;(Hyd 2c )2N and C 1-5 wherein 0, 1 or 2 carbon atoms in the hydrocarbon group are replaced by heteroatoms selected from N, O and S, and the hydrocarbon group is optionally substituted by one or more fluorine atoms or by a 5-membered or 6-membered heteroaryl group containing 1 or 2 heteroatom ring members selected from N and O;

[0084] R c Selected from hydrogen and C 1-4 non-aromatic hydrocarbon groups; and

[0085] R 7 Independently selected from R 4 .

[0086] In formulae (1A) and (1B), Z is a 5-membered heteroaryl ring comprising one or two nitrogen ring members and optionally one further heteroatom ring member selected from N and O.

[0087] It will be appreciated that when the 5-membered heteroaryl ring Z contains a second heteroatom ring member, for example when it is pyrazole or isoxazole, R 2 and R 3 Thus, in each of the above and following aspects and embodiments, when the 5-membered heteroaryl ring is not pyrrole, the definition is deemed to include where R 2 and R 3 Compounds in which one or both of these are absent.

[0088] Specific and preferred aspects and embodiments of the invention are set out below in Embodiments 1.2 to 1.191.

[0089] 1.2 The composition of matter according to embodiment 1.1, with the proviso that the composition of matter is not a composition of matter comprising:

[0090] (i) Atropisomers, where R 1 is a methyl group, and R 4 is a 4-cyano or 4-carbamoyl group;

[0091] (ii) Atropisomers, where R 6 is hydroxy, methoxymethyl or unsubstituted or fluorine-substituted C 1-8 Alkoxy (e.g., trifluoromethoxy);

[0092] (iii) an atropisomer wherein ring Z is an isoxazole ring and Ar 1is an unsubstituted 4-pyridyl group attached to the isoxazole 3 position; and R 2 and R 3 None of them exist; or

[0093] (iv) an atropisomer wherein Z is an isoxazole ring and R 4 is an azetidin-4-yloxy group.

[0094] 1.3 A composition of matter according to embodiment 1.1 or embodiment 1.2 which is not a pyrrole substituted at each of the 1, 2 and 3 positions with a phenyl or pyridyl ring.

[0095] 1.4 A composition of matter according to any one of Embodiments 1.1 to 1.3, wherein the ring Z is not a 1,2,3-trisubstituted pyrrole ring.

[0096] 1.5 A composition of matter according to any one of Embodiments 1.1 to 1.4, wherein the ring Z is not an imidazole ring.

[0097] 1.6 A composition of matter according to any one of Embodiments 1.1 to 1.5, wherein the ring Z is not a 1,2,4 triazole ring.

[0098] 1.7 A composition of matter according to any one of embodiments 1.1 to 1.6, wherein Z is a heteroaryl ring comprising a nitrogen ring member and optionally one further heteroatom ring member selected from N and O; or Z is a triazole ring.

[0099] 1.8 A composition of matter according to embodiment 1.7, wherein Z is selected from pyrrole, isoxazole, imidazole, pyrazole and triazole rings.

[0100] 1.9 A composition of matter according to embodiment 1.8, wherein Z is selected from pyrrole, pyrazole and isoxazole rings.

[0101] 1.10 A composition of matter according to Embodiment 1.9 wherein Z is a pyrrole ring.

[0102] 1.11 A composition of matter according to Embodiment 1.10 wherein Ring X is attached to the nitrogen atom of the pyrrole ring.

[0103] 1.12 A composition of matter according to Embodiment 1.9 wherein Z is a pyrazole ring.

[0104] 1.13 A composition of matter according to Embodiment 1.12 wherein Ring X is attached to a carbon atom of the pyrazole ring.

[0105] 1.14 A composition of matter according to Embodiment 1.12 or Embodiment 1.13 wherein Ring Y is attached to a carbon atom of the pyrazole ring.

[0106] 1.15 A composition of matter according to Embodiment 1.12 or Embodiment 1.13 wherein Ring Y is attached to the nitrogen atom of the pyrazole ring.

[0107] 1.16 A composition of matter according to any one of embodiments 1.12 to 1.15, wherein Ar 1 is attached to the carbon atom of the pyrazole ring.

[0108] 1.17 A composition of matter according to Embodiment 1.9 wherein Z is an isoxazole ring.

[0109] 1.18 A composition of matter according to Embodiment 1.17 wherein Ring X is attached to the 4-position of the isoxazole ring.

[0110] 1.19 A composition of matter according to Embodiment 1.17 or Embodiment 1.18 wherein Ring Y is attached to the 5-position of the isoxazole ring.

[0111] 1.20 A composition of matter according to any one of Embodiments 1.17 to 1.19, wherein Ar 1 Attached to the 3-position of the isoxazole ring.

[0112] 1.21 A composition of matter according to any one of Embodiments 1.1 to 1.20, wherein the ring X is a benzene, pyridine or pyrimidine ring.

[0113] 1.22 A composition of matter according to Embodiment 1.21 wherein the ring X is a benzene ring or a pyridine ring.

[0114] 1.23 A composition of matter according to Embodiment 1.22 wherein the ring X is a benzene ring.

[0115] 1.24 A composition of matter according to Embodiment 1.22 wherein the ring X is a pyridine ring.

[0116] 1.25 A composition of matter according to any one of Embodiments 1.21, 1.22 and 1.24 wherein the pyridine ring is a 2-pyridine ring.

[0117] 1.26 A composition of matter according to any one of Embodiments 1.21, 1.22 and 1.24 wherein the pyridine ring is a 3-pyridine ring.

[0118] 1.27 A composition of matter according to any one of Embodiments 1.21, 1.22 and 1.24 wherein the pyridine ring is a 4-pyridine ring.

[0119] 1.28 A composition of matter according to any one of Embodiments 1.21, 1.22 and 1.24, wherein the pyridine ring is a 2-pyridine or 3-pyridine ring.

[0120] 1.29 A composition of matter according to any one of embodiments 1.1 to 1.28, wherein R 1 Selected from:

[0121] -chlorine;

[0122] -bromine;

[0123] -hydroxyl group;

[0124] - cyano;

[0125] -carboxyl;

[0126] -amino group;

[0127] --(Hyd 2 )NH;

[0128] -(Hyd 2 )2N;

[0129] -C 1-5 A hydrocarbon group, wherein 0, 1 or 2 carbons in the hydrocarbon group are replaced by heteroatoms selected from N, O and S, and the hydrocarbon group is optionally substituted by one or more fluorine atoms.

[0130] 1.30 The composition of matter according to embodiment 1.29, wherein R 1 Selected from:

[0131] -chlorine;

[0132] -bromine;

[0133] -hydroxyl group;

[0134] -carboxyl;

[0135] -amino group;

[0136] -Methylamino;

[0137] -dimethylamino;

[0138] -C 1-5 A hydrocarbon group, wherein 0, 1 or 2 carbons in the hydrocarbon group are replaced by heteroatoms selected from N, O and S, and the hydrocarbon group is optionally substituted by one or more fluorine atoms.

[0139] 1.31 A composition of matter according to embodiment 1.29, wherein R 1 Selected from:

[0140] -chlorine;

[0141] -bromine;

[0142] -hydroxyl group;

[0143] -amino group;

[0144] -C 1-5 A hydrocarbon group, wherein 0, 1 or 2 carbons in the hydrocarbon group are replaced by heteroatoms selected from N, O and S, and the hydrocarbon group is optionally substituted by one or more fluorine atoms.

[0145] 1.32 A composition of matter according to embodiment 1.29, wherein R 1 Selected from:

[0146] -hydroxyl group;

[0147] -amino group; and

[0148] -C 1-5 A hydrocarbon group, wherein 0, 1 or 2 carbons in the hydrocarbon group are replaced by heteroatoms selected from N, O and S, and the hydrocarbon group is optionally substituted by one or more fluorine atoms.

[0149] 1.33 A composition of matter according to embodiment 1.29, wherein R 1 Selected from:

[0150] -hydroxyl group;

[0151] -amino group; and

[0152] -C 1-4 A hydrocarbon group, wherein 0, 1 or 2 carbons in the hydrocarbon group are replaced by heteroatoms selected from N and O, and the hydrocarbon group is optionally substituted by one or more fluorine atoms.

[0153] 1.34 A composition of matter according to embodiment 1.29, wherein R 1 Selected from saturated C 1-4 A hydrocarbon group, wherein 0, 1 or 2 carbons in the hydrocarbon group are replaced by heteroatoms selected from N and O, and the hydrocarbon group is optionally substituted by one or more fluorine atoms.

[0154] 1.35 A composition of matter according to embodiment 1.29, wherein R 1 Selected from saturated C 1-4 A hydrocarbon group, wherein 0 or 1 carbon in the hydrocarbon group is replaced by a heteroatom selected from N and O, and the hydrocarbon group is optionally substituted by one or more fluorine atoms.

[0155] 1.36 A composition of matter according to embodiment 1.29, wherein R 1 Selected from C 1-4An alkyl group, wherein 0 or 1 carbon in the alkyl group is replaced by a heteroatom selected from N and O, and the hydrocarbon group is optionally substituted by one or more fluorine atoms.

[0156] 1.37 A composition of matter according to embodiment 1.29, wherein R 1 is selected from hydroxyl; carboxyl; amino; C optionally substituted by one or more fluorine atoms 1-4 An alkyl group; a C optionally substituted by one or more fluorine atoms 1-3 Alkoxy groups; (dimethylamino)methyl and (methoxy)methyl.

[0157] 1.38 A composition of matter according to embodiment 1.29, wherein R 1 Selected from hydroxy; carboxyl; amino; trifluoromethyl; (dimethylamino)methyl and (methoxy)methyl.

[0158] 1.39 A composition of matter according to embodiment 1.29, wherein R 1 is C optionally substituted by one or more fluorine atoms 1-4 or a C optionally substituted by one or more fluorine atoms 1-3 Alkoxy groups.

[0159] 1.40 The composition of matter according to embodiment 1.29, wherein R 1 is C substituted with one or more fluorine atoms 1-4 Alkyl group.

[0160] 1.41 A composition of matter according to Embodiment 1.29, wherein R 1 is C substituted with one or more fluorine atoms 1-2 Alkyl group.

[0161] 1.42 A composition of matter according to embodiment 1.41, wherein R 1 It is a methyl group substituted by two or three fluorine atoms.

[0162] 1.43 A composition of matter according to embodiment 1.42, wherein R 1 It is trifluoromethyl.

[0163] 1.44 A composition of matter according to Embodiment 1.29, wherein R 1 is selected from hydrogen, trifluoromethyl, trifluoromethoxy, difluoromethyl or difluoromethoxy, hydroxy, amino, carboxyl, (dimethylamino)methyl and (methoxy)methyl.

[0164] 1.45 A composition of matter according to embodiment 1.29, wherein R 1Selected from trifluoromethyl; hydroxy; amino; (dimethylamino)methyl and (methoxy)methyl.

[0165] 1.46 A composition of matter according to any one of Embodiments 1.1 to 1.45 wherein m is 0 or 1.

[0166] 1.47 A composition of matter according to any one of Embodiments 1.1 to 1.45 wherein m is 0.

[0167] 1.48 A composition of matter according to any one of Embodiments 1.1 to 1.45 wherein m is 1.

[0168] 1.49 A composition of matter according to any one of Embodiments 1.1 to 1.45 wherein m is 2.

[0169] 1.50 A composition of matter according to any one of embodiments 1.1 to 1.46, 1.48 and 1.49, wherein R 4 Selected from:

[0170] -fluorine;

[0171] -chlorine;

[0172] -bromine;

[0173] - cyano; and

[0174] -C 1-5 A hydrocarbon group, wherein 0, 1 or 2 carbons in the hydrocarbon group are replaced by heteroatoms selected from N, O and S, and the hydrocarbon group is optionally substituted by one or more fluorine atoms.

[0175] 1.51 A composition of matter according to embodiment 1.50, wherein R 4 Selected from:

[0176] -fluorine;

[0177] -chlorine;

[0178] -bromine;

[0179] - cyano; and

[0180] -C 1-4 A hydrocarbon group, wherein 0 or 1 carbon in the hydrocarbon group is replaced by a heteroatom selected from N, O and S, and the hydrocarbon group is optionally substituted by one or more fluorine atoms.

[0181] 1.52 A composition of matter according to embodiment 1.51, wherein R 4 Selected from:

[0182] -fluorine;

[0183] -chlorine;

[0184] -bromine;

[0185] - cyano; and

[0186] C 1-4 An alkyl group, wherein 0 or 1 carbon in the alkyl group is replaced by a heteroatom selected from N and O, and the alkyl group is optionally substituted by one or more fluorine atoms.

[0187] 1.53 A composition of matter according to embodiment 1.52, wherein R 4 Selected from:

[0188] -fluorine;

[0189] -chlorine;

[0190] - bromine; and

[0191] -C 1-4 An alkyl group, wherein 0 or 1 carbon in the alkyl group is replaced by a heteroatom O, and the alkyl group is optionally substituted by one or more fluorine atoms.

[0192] 1.54 A composition of matter according to embodiment 1.53, wherein R 4 Selected from fluorine; chlorine; bromine and C 1-4 alkyl.

[0193] 1.55 A composition of matter according to embodiment 1.54, wherein R 4 selected from fluorine; chlorine; and C 1-4 alkyl.

[0194] 1.56 A composition of matter according to any one of embodiments 1.1 to 1.55, wherein R 2 Selected from hydrogen and saturated C 1-4 Hydrocarbon group.

[0195] 1.57 A composition of matter according to embodiment 1.56, wherein R 2 Selected from hydrogen; C 1-4 Alkyl; cyclopropyl and cyclopropylmethyl.

[0196] 1.58 A composition of matter according to embodiment 1.57, wherein R 2 Selected from hydrogen; C 1-3 Alkyl and cyclopropyl.

[0197] 1.59 A composition of matter according to embodiment 1.58, wherein R 2 Selected from hydrogen; methyl and ethyl.

[0198] 1.60 A composition of matter according to Embodiment 1.59, wherein R2 is hydrogen or methyl.

[0199] 1.61 A composition of matter according to Embodiment 1.60, wherein R 2 It's hydrogen.

[0200] 1.62 A composition of matter according to any one of Embodiments 1.1 to 1.61, wherein R 3 Selected from hydrogen and saturated C 1-4 Hydrocarbon group.

[0201] 1.63 The composition of matter according to embodiment 1.62, wherein R 3 Selected from hydrogen; C 1-4 Alkyl; cyclopropyl and cyclopropylmethyl.

[0202] 1.64 A composition of matter according to Embodiment 1.63, wherein R 3 Selected from hydrogen; C 1-3 Alkyl and cyclopropyl.

[0203] 1.65 A composition of matter according to embodiment 1.64, wherein R 3 Selected from hydrogen; methyl and ethyl.

[0204] 1.66 A composition of matter according to embodiment 1.65, wherein R 3 is hydrogen or methyl.

[0205] 1.67 A composition of matter according to Embodiment 1.66, wherein R 3 It's hydrogen.

[0206] 1.68 A composition of matter according to any one of embodiments 1.1 to 1.67, wherein Ar 1 is a monocyclic aromatic ring selected from the group consisting of benzene; pyridine; pyrimidine; thiophene; and furan; each of the monocyclic aromatic rings is optionally substituted by one or more substituents R 5 replace.

[0207] 1.69 A composition of matter according to embodiment 1.68, wherein Ar 1 is a monocyclic aromatic ring selected from the group consisting of benzene; pyridine and pyrimidine; each of the monocyclic aromatic rings is optionally substituted by one or more substituents R 5 replace.

[0208] 1.70 A composition of matter according to embodiment 1.69, wherein Ar 1 is a monocyclic aromatic ring selected from the group consisting of benzene and pyridine; each of the monocyclic aromatic rings is optionally substituted by one or more substituents R 5 replace.

[0209] 1.71 A composition of matter according to embodiment 1.70, wherein Ar 1 is optionally substituted by one or more substituents R 5 Substituted benzene ring.

[0210] 1.72 A composition of matter according to embodiment 1.70, wherein Ar 1 is optionally substituted by one or more substituents R 5 Substituted pyridine ring.

[0211] 1.73 A composition of matter according to any one of embodiments 1.1 to 1.72, wherein the monocyclic aromatic ring Ar 1 is unsubstituted or substituted by 1, 2 or 3 groups R 5 replace.

[0212] 1.74 A composition of matter according to embodiment 1.73, wherein the monocyclic aromatic ring Ar 1 is unsubstituted or substituted with 1 or 2 substituents R 5 replace.

[0213] 1.75 A composition of matter according to embodiment 1.74, wherein the monocyclic aromatic ring Ar 1 is unsubstituted or substituted with 1 substituent R 5 replace.

[0214] 1.76 A composition of matter according to embodiment 1.75, wherein the monocyclic aromatic ring Ar 1 Substituted by 1 group R 5 replace.

[0215] 1.77 A composition of matter according to embodiment 1.75, wherein the monocyclic aromatic ring Ar 1 is unsubstituted.

[0216] 1.78 A composition of matter according to embodiment 1.74, wherein the monocyclic aromatic ring Ar 1 Substituted by 2 groups R 5 replace.

[0217] 1.79 A composition of matter according to any one of embodiments 1.1 to 1.76 and 1.78, wherein R 5 Selected from halogen family; O-Ar 2 ; Cyano, Hyd 1b -SO2- and C 1-8 A hydrocarbon group, wherein 0, 1 or 2 carbons but not all carbons in the hydrocarbon group are optionally replaced by heteroatoms selected from N, O and S, and wherein the hydrocarbon group is optionally substituted with one or more fluorine atoms.

[0218] 1.80 A composition of matter according to any one of embodiments 1.1 to 1.76, 1.78 and 1.79, wherein Ar 2 is a phenyl or pyridyl group optionally substituted by 1 or 2 substituents selected from fluorine, chlorine, cyano and trifluoromethyl.

[0219] 1.81 A composition of matter according to embodiment 1.80, wherein Ar 2 is a phenyl group optionally substituted by 1 or 2 substituents selected from fluorine, chlorine, cyano and trifluoromethyl.

[0220] 1.82 A composition of matter according to any one of embodiments 1.1 to 1.76 and 1.78 to 1.80, wherein Hyd 1b It is saturated C 1-4 Hydrocarbon group.

[0221] 1.83 A composition of matter according to embodiment 1.82, wherein Hyd 1b Selected from C 1-4 Alkyl; cyclopropyl and cyclopropylmethyl.

[0222] 1.84 A composition of matter according to embodiment 1.78, wherein Hyd 1b Selected from methyl; ethyl; propyl; cyclopropyl and cyclopropylmethyl.

[0223] 1.85 A composition of matter according to embodiment 1.79, wherein Hyd 1b Selected from methyl; ethyl; propyl and cyclopropyl.

[0224] 1.86 A composition of matter according to embodiment 1.80, wherein Hyd 1b Selected from methyl and ethyl.

[0225] 1.87 A composition of matter according to Embodiment 1.81, wherein Hyd 1b It's methyl.

[0226] 1.88 A composition of matter according to any one of embodiments 1.1 to 1.76 and 1.78, wherein R 5 Selected from bromine; fluorine; chlorine; cyano; phenoxy; C 1-4 Alkylsulfonyl; C 1-4 Alkoxy and C 1-4 Alkyl, wherein the C 1-4 Alkoxy and C 1-4 The alkyl groups are each optionally substituted with one or more fluorine atoms.

[0227] 1.89 A composition of matter according to embodiment 1.88, wherein R 5is selected from the group consisting of bromo; fluoro; chloro; cyano; phenoxy; methylsulfonyl; methyl; ethyl; isopropyl; difluoromethyl; trifluoromethyl; methoxy; difluoromethoxy; and trifluoromethoxy.

[0228] 1.90 A composition of matter according to embodiment 1.89, wherein R 5 is selected from bromo; fluoro; chloro; cyano; phenoxy; methylsulfonyl; and isopropyl.

[0229] 1.90A A composition of matter according to any one of Embodiments 1.1 to 1.90, wherein R 5 Located in the monocyclic aromatic ring Ar 1 The counterpoint on.

[0230] 1.91 A composition of matter according to embodiment 1.90 or embodiment 1.91, wherein R 5 is selected from bromine; fluorine; chlorine; and cyano.

[0231] 1.92 A composition of matter according to embodiment 1.91, wherein R 5 Selected from fluorine, chlorine and cyano.

[0232] 1.93 A composition of matter according to embodiment 1.92, wherein R 5 It's cyano.

[0233] 1.94 A composition of matter according to embodiment 1.93, wherein Ar 1 It is 4-cyanophenyl.

[0234] 1.95 A composition of matter according to embodiment 1.92, wherein R 5 It's chlorine.

[0235] 1.96 A composition of matter according to embodiment 1.95, wherein Ar 1 It is 4-chlorophenyl.

[0236] 1.97 A composition of matter according to embodiment 1.92, wherein R 5 It's fluorine.

[0237] 1.98 A composition of matter according to embodiment 1.97, wherein Ar 1 It is 4-fluorophenyl.

[0238] 1.99 A composition of matter according to any one of Embodiments 1.1 to 1.98 wherein the ring Y is a benzene, pyridine, pyrimidine, furan, thiophene or pyrrole ring.

[0239] 1.100 A composition of matter according to Embodiment 1.99 wherein the ring Y is a) a benzene ring, b) a pyridine ring or c) a thiophene ring.

[0240] 1.101 A composition of matter according to Embodiment 1.100 wherein Ring Y is a benzene ring.

[0241] 1.102 A composition of matter according to Embodiment 1.100 wherein Ring Y is a pyridine ring.

[0242] 1.103 A composition of matter according to any one of Embodiments 1.1 to 1.102, wherein R 6 It is a group Q 1 -R a -R b .

[0243] 1.104 A composition of matter according to any one of Embodiments 1.1 to 1.103, wherein Q 1 With the formula (CR p R q ) r , where r is 0, 1, 2, 3 or 4, and R p and R q are independently selected from hydrogen and methyl or R p and R q Together with the carbon atoms to which they are attached, they form a 3- or 4-membered saturated cyclic hydrocarbon ring, provided that Q 1 The total number of carbon atoms does not exceed 6.

[0244] 1.105 A composition of matter according to any one of Embodiments 1.1 to 1.104, wherein Q 1 Absent or selected from CH2, CH(CH3), C(CH3)2, cyclopropane-1,1-diyl and cyclobutane-1,1-diyl.

[0245] 1.106 A composition of matter according to Embodiment 1.105, wherein Q 1 Does not exist.

[0246] 1.107 A composition of matter according to Embodiment 1.105, wherein Q 1 It is a -CH2- group.

[0247] 1.108 A composition of matter according to any one of Embodiments 1.1 to 1.107, wherein R a Not present or selected from O; C(O); C(O)O; CONR c ; N(R c )CO; N(R c )CONR c NR c ; and SO2.

[0248] 1.109 A composition of matter according to embodiment 1.108, wherein R a Not present or selected from O; CONR c ; N(R c )CO; N(R c )CONR c NR c and SO2.

[0249] 1.110 A composition of matter according to embodiment 1.108, wherein R a It is CONR c .

[0250] 1.111 A composition of matter according to Embodiment 1.108, wherein R a is N(R c )CO.

[0251] 1.112 A composition of matter according to embodiment 1.108, wherein R a YesNR c .

[0252] 1.113 A composition of matter according to Embodiment 1.108, wherein R a Does not exist.

[0253] 1.114 A composition of matter according to embodiment 1.108, wherein R a It's O.

[0254] 1.115 A composition of matter according to Embodiment 1.108, wherein R a It is C(O).

[0255] 1.116 A composition of matter according to embodiment 1.108, wherein R a It is C(O)O.

[0256] 1.117 A composition of matter according to Embodiment 1.108, wherein R a It's SO2.

[0257] 1.118 A composition of matter according to any one of Embodiments 1.1 to 1.117, wherein R b Selected from:

[0258] -C 1-8 A non-aromatic hydrocarbon group, wherein 0, 1 or 2 carbon atoms in the hydrocarbon group, but not all carbon atoms, are replaced by heteroatoms selected from N and O, and the C 1-8 The non-aromatic hydrocarbon group is optionally substituted by one or more selected from fluorine and the group Cyc 1 substituted with a substituent; and

[0259] -Cyc Group 1 ;

[0260] The premise is that when R a is C(O)O or CONR c When R b Additionally selected from hydrogen.

[0261] 1.119 A composition of matter according to Embodiment 1.118, wherein R b Selected from:

[0262] -C 1-8 A non-aromatic hydrocarbon group, wherein 0, 1 or 2 carbon atoms in the hydrocarbon group are replaced by heteroatoms selected from N and O, and the C 1-8 The non-aromatic hydrocarbon group is optionally substituted by one or more selected from fluorine and the group Cyc 1 substituted with a substituent; and

[0263] -Cyc Group 1 .

[0264] 1.120 A composition of matter according to embodiment 1.119, wherein R b Selected from:

[0265] -C 1-8 A non-aromatic hydrocarbon group, wherein 0 or 1 carbon atom in the hydrocarbon group, but not all carbon atoms, is replaced by a heteroatom selected from N and O, and the C 1-8 The non-aromatic hydrocarbon group is optionally substituted by one or more selected from fluorine and the group Cyc 1 substituted with a substituent; and

[0266] -Cyc Group 1 .

[0267] 1.121 A composition of matter according to Embodiment 1.120, wherein R b Selected from:

[0268] -C 1-8 A non-aromatic hydrocarbon group, wherein 0 or 1 carbon atom in the hydrocarbon group, but not all carbon atoms, is replaced by a heteroatom selected from N and O, and the C 1-8 The non-aromatic hydrocarbon group is optionally replaced by a Cyc group. 1 Replacement; and

[0269] -Cyc Group 1 .

[0270] 1.122 A composition of matter according to Embodiment 1.121, wherein R b Selected from:

[0271] -C1-8 A non-aromatic hydrocarbon group, wherein one carbon atom in the hydrocarbon group is replaced by a heteroatom selected from N and O; and

[0272] -Cyc Group 1 .

[0273] 1.123 A composition of matter according to embodiment 1.122, wherein R b Selected from:

[0274] -C 1-8 A non-aromatic hydrocarbon group, wherein one carbon atom in the hydrocarbon group is replaced by a heteroatom N; and

[0275] -Cyc Group 1 .

[0276] 1.124 A composition of matter according to any one of Embodiments 1.1 to 1.117, wherein R b It is C 1-8 A non-aromatic hydrocarbon group, wherein 0, 1 or 2 carbon atoms in the hydrocarbon group, but not all carbon atoms, are replaced by heteroatoms selected from N and O, and the C 1-8 The non-aromatic hydrocarbon group is optionally substituted by one or more selected from fluorine and the group Cyc 1 substituted by a substituent.

[0277] 1.125 A composition of matter according to Embodiment 1.124, wherein R b Selected from:

[0278] -C 1-8 A non-aromatic hydrocarbon group, wherein 0 or 1 carbon atom in the hydrocarbon group, but not all carbon atoms, is replaced by a heteroatom selected from N and O, and the C 1-8 The non-aromatic hydrocarbon group is optionally substituted by one or more selected from fluorine and the group Cyc 1 substituted by a substituent.

[0279] 1.126 A composition of matter according to Embodiment 1.125, wherein R b Selected from:

[0280] -C 1-8 A non-aromatic hydrocarbon group, wherein 0 or 1 carbon atom in the hydrocarbon group, but not all carbon atoms, is replaced by a heteroatom selected from N and O, and the C 1-8 The non-aromatic hydrocarbon group is optionally replaced by a Cyc group. 1 replace.

[0281] 1.127 A composition of matter according to Embodiment 1.126, wherein R b Selected from:

[0282] -C1-8 A non-aromatic hydrocarbon group, wherein one carbon atom in the hydrocarbon group is replaced by a heteroatom selected from N and O.

[0283] 1.128 A composition of matter according to any one of Embodiments 1.1 to 1.127, wherein R b Selected from:

[0284] C 1-8 A non-aromatic hydrocarbon group, wherein one carbon atom in the hydrocarbon group is replaced by a nitrogen heteroatom.

[0285] 1.129 A composition of matter according to any one of embodiments 1.118 to 1.128, wherein R b Selected from:

[0286] C 1-8 A non-aromatic hydrocarbon group wherein a carbon atom in the hydrocarbon group is replaced by a nitrogen heteroatom to form a terminal dimethylamino group.

[0287] 1.130 A composition of matter according to any one of Embodiments 1.118 to 1.129 wherein the non-aromatic hydrocarbon group is acyclic.

[0288] 1.131 A composition of matter according to any one of Embodiments 1.118 to 1.130 wherein the non-aromatic hydrocarbon group is saturated.

[0289] 1.132 A composition of matter according to any one of Embodiments 1.118 to 1.131 wherein the non-aromatic hydrocarbon group contains 1 to 6 carbon atoms.

[0290] 1.133 A composition of matter according to any one of Embodiments 1.118 to 1.132 wherein the non-aromatic hydrocarbon group contains 1 to 5 carbon atoms.

[0291] 1.134 A composition of matter according to any one of Embodiments 1.118 to 1.133 wherein the non-aromatic hydrocarbon group contains 3 to 5 carbon atoms.

[0292] 1.135 A composition of matter according to any one of Embodiments 1.1 to 1.126, wherein R b Is or contains the group Cyc 1 .

[0293] 1.136 A composition of matter according to embodiment 1.128, wherein R b It is the group Cyc 1 .

[0294] 1.137 A composition of matter according to Embodiment 1.136 wherein Cyc1 is a non-aromatic 4-7 membered carbocyclic or heterocyclic group containing 0, 1 or 2 heteroatom ring members selected from N and O and optionally substituted by one or more selected from hydroxy; amino; mono-C 1-4 Alkylamino; di-C 1-4 Alkylamino; and C 1-5 Substituents of a saturated hydrocarbon group are substituted wherein 0 or 1 carbon, but not all carbons, in the hydrocarbon group are replaced by heteroatoms selected from N and O.

[0295] 1.138 A composition of matter according to Embodiment 1.137, wherein Cyc 1 is a non-aromatic 4-7 membered heterocyclic group comprising a nitrogen ring member and a second heteroatom ring member optionally selected from N and O; the non-aromatic 4-7 membered heterocyclic group is optionally substituted by one or more substituents selected from hydroxyl; amino; mono-C 1-4 Alkylamino; di-C 1-4 Alkylamino; and C 1-4 A saturated hydrocarbon group wherein 0 or 1 carbon, but not all carbons, of the hydrocarbon group are replaced by heteroatoms selected from N and O.

[0296] 1.139 A composition of matter according to Embodiment 1.138, wherein Cyc 1 is a non-aromatic 5-6 membered heterocyclic group comprising a nitrogen ring member and a second heteroatom ring member optionally selected from N and O; the non-aromatic 5-6 membered heterocyclic group is optionally substituted by one or more substituents selected from hydroxyl; amino; mono-C 1-4 Alkylamino; di-C 1-4 Alkylamino; and C 1-4 A saturated hydrocarbon group wherein 0 or 1 carbon, but not all carbons, of the hydrocarbon group are replaced by heteroatoms selected from N and O.

[0297] 1.140 A composition of matter according to Embodiment 1.139, wherein Cyc 1 is a non-aromatic 5-6 membered heterocyclic group comprising a nitrogen ring member and a second heteroatom ring member optionally selected from N and O; the non-aromatic 5-6 membered heterocyclic group is optionally substituted by one or more substituents selected from hydroxyl; amino; mono- C1-2 Alkylamino; di-C 1-2 Alkylamino; and C 1-4 An alkyl group wherein 0 or 1 carbon, but not all carbons, of the alkyl group are replaced by heteroatoms selected from N and O.

[0298] 1.141 A composition of matter according to any one of embodiments 1.1 to 1.126 and 1.135 to 1.140, wherein Cyc 1 It is a saturated ring.

[0299] 1.142 A composition of matter according to Embodiment 1.141 wherein Cyc 1 selected from pyrrolidine; piperidine; and piperazine; each of which is optionally replaced by one or more selected from hydroxy; amino; mono-C 1-2 Alkylamino; di-C 1-2 Alkylamino; and C 1-4 Substituents of an alkyl group wherein 0 or 1 carbon, but not all carbons, in the alkyl group are replaced by heteroatoms selected from N and O.

[0300] 1.143 A composition of matter according to any one of embodiments 1.1 to 1.112 and 1.118 to 1.142, wherein R c is selected from the group consisting of hydrogen, methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclopropylmethyl, butyl, isobutyl and cyclobutyl.

[0301] 1.144 A composition of matter according to embodiment 1.143, wherein R c is selected from hydrogen and methyl.

[0302] 1.145 A composition of matter according to Embodiment 1.144, wherein R c It's hydrogen.

[0303] 1.146 A composition of matter according to embodiment 1.144, wherein R c It's methyl.

[0304] 1.147 A composition of matter according to any one of Embodiments 1.1 to 1.103, wherein R 6 is selected from groups A to AL in the following table.

[0305]

[0306]

[0307]

[0308] 1.148 A composition of matter according to embodiment 1.147, wherein R 6 Selected from the groups A and Q.

[0309] 1.149 A composition of matter according to embodiment 1.148, wherein R 6 It is group A.

[0310] 1.149A A composition of matter according to any one of Embodiments 1.1 to 1.149 wherein n is 0 or 1.

[0311] 1.149B A composition of matter according to Embodiment 1.149A wherein n is 0.

[0312] 1.149C A composition of matter according to Embodiment 1.149A wherein n is 1.

[0313] 1.149D A composition of matter according to any one of embodiments 1.1 to 1.149A and 1.149C, wherein R 7 is selected from fluorine, chlorine and methoxy.

[0314] 1.149E A composition of matter according to Embodiment 1.149D wherein R 7 Selected from chlorine and methoxy.

[0315] 1.149F A composition of matter according to Embodiment 1.149D wherein n is 1 and R 7 It's chlorine.

[0316] 1.149G A composition of matter according to Embodiment 1.149D wherein n is 1 and R 7 It's a methoxy group.

[0317] 1.150 A composition of matter according to any one of Embodiments 1.1 to 1.149, wherein: (i) when Y is a six-membered ring, R 6 or (ii) when Y is a five-membered ring, R 6 is attached to ring Y at a position that is not adjacent to a ring member of Y that is attached to ring Z.

[0318] 1.151 A composition of matter according to embodiment 1.150, wherein Y is a six-membered ring and R 6 Connect in between or opposite positions.

[0319] 1.152 A composition of matter according to embodiment 1.151, wherein Y is a six-membered ring and R 6 Connect in between.

[0320] 1.153 A composition of matter according to embodiment 1.151 wherein Y is a six-membered ring and R 6 Connect in its position.

[0321] 1.154 A composition of matter consisting of at least 90% by weight of an atropisomer of formula (1A) and 0-10% by weight of an atropisomer of formula (1B); wherein the atropisomer of formula (1A) and the atropisomer of formula (1B) are represented by:

[0322]

[0323] or a pharmaceutically acceptable salt or tautomer thereof, wherein R 1 To R 7 ,Ar 1 , m, n, X, Y and Z are as defined in any one of Embodiments 1.1 to 1.153.

[0324] 1.155 A composition of matter according to Embodiment 1.154 consisting of at least 95% by weight of the atropisomer (1A) or a salt or tautomer thereof and 0-5% by weight of the atropisomer of formula (1B) or a salt or tautomer thereof.

[0325] 1.156 A composition of matter according to Embodiment 1.154 consisting of at least 96% by weight of the atropisomer (1A) or a salt or tautomer thereof and 0-4% by weight of the atropisomer of formula (1B) or a salt or tautomer thereof.

[0326] 1.157 A composition of matter according to Embodiment 1.154 consisting of at least 97% by weight of the atropisomer (1A) or a salt or tautomer thereof and 0-3% by weight of the atropisomer of formula (1B) or a salt or tautomer thereof.

[0327] 1.158 A composition of matter according to Embodiment 1.154 consisting of at least 98% by weight of the atropisomer (1A) or a salt or tautomer thereof and 0-2% by weight of the atropisomer of formula (1B) or a salt or tautomer thereof.

[0328] 1.159 A composition of matter according to Embodiment 1.154 consisting of at least 99% by weight of the atropisomer (1A) or a salt or tautomer thereof and 0-1% by weight of the atropisomer of formula (1B) or a salt or tautomer thereof.

[0329] 1.160 A composition of matter according to Embodiment 1.154 consisting of at least 99.5% by weight of the atropisomer (1A) or a salt or tautomer thereof and 0-0.5% by weight of the atropisomer of formula (1B) or a salt or tautomer thereof.

[0330] 1.161 A composition of matter consisting of at least 90% by weight of an atropisomer (1B) and 0-10% by weight of an atropisomer of formula (1A); wherein the atropisomer of formula (1A) and the atropisomer of formula (1B) are represented by:

[0331]

[0332] or a pharmaceutically acceptable salt or tautomer thereof, wherein R 1 To R7 ,Ar 1 , m, n, X, Y and Z are as defined in any one of Embodiments 1.1 to 1.153.

[0333] 1.162 A composition of matter according to Embodiment 1.161 consisting of at least 95% by weight of the atropisomer (1B) or a salt or tautomer thereof and 0-5% by weight of the atropisomer of formula (1A) or a salt or tautomer thereof.

[0334] 1.163 A composition of matter according to Embodiment 1.161 consisting of at least 96% by weight of the atropisomer (1B) or a salt or tautomer thereof and 0-4% by weight of the atropisomer of formula (1A) or a salt or tautomer thereof.

[0335] 1.164 A composition of matter according to Embodiment 1.161 consisting of at least 97% by weight of the atropisomer (1B) or a salt or tautomer thereof and 0-3% by weight of the atropisomer of formula (1A) or a salt or tautomer thereof.

[0336] 1.165 A composition of matter according to Embodiment 1.161 consisting of at least 98% by weight of the atropisomer (1B) or a salt or tautomer thereof and 0-2% by weight of the atropisomer of formula (1A) or a salt or tautomer thereof.

[0337] 1.166 A composition of matter according to Embodiment 1.161 consisting of at least 99% by weight of the atropisomer (1B) or a salt or tautomer thereof and 0-1% by weight of the atropisomer of formula (1A) or a salt or tautomer thereof.

[0338] 1.167 A composition of matter according to Embodiment 1.161 consisting of at least 99.5% by weight of the atropisomer (1B) or a salt or tautomer thereof and 0-0.5% by weight of the atropisomer of formula (1A) or a salt or tautomer thereof.

[0339] 1.168 A composition of matter:

[0340] (i) consists of at least 90% by weight of the atropisomer (2A) and 0-10% by weight of the atropisomer of formula (2B); or

[0341] (ii) consists of at least 90% by weight of the atropisomer (2B) and 0-10% by weight of the atropisomer of formula (2A);

[0342] The atropisomer of formula (2A) and the atropisomer of formula (2B) are represented by the following:

[0343]

[0344] or a pharmaceutically acceptable salt or tautomer thereof, wherein R 1 , R 2 , R 3 , R 4 , R 6 , R 7 ,Ar 1 , X and Y are as defined in any one of Embodiments 1.1, 1.2, 1.8, 1.10, 1.11 and 1.21 to 1.153.

[0345] 1.169 A composition of matter according to Embodiment 1.168 consisting of at least 95% by weight of the atropisomer (2A) or a salt or tautomer thereof and 0-5% by weight of the atropisomer of formula (2B) or a salt or tautomer thereof.

[0346] 1.170 A composition of matter according to Embodiment 1.168 consisting of at least 96% by weight of the atropisomer (2A) or a salt or tautomer thereof and 0-4% by weight of the atropisomer of formula (2B) or a salt or tautomer thereof.

[0347] 1.171 A composition of matter according to Embodiment 1.168 consisting of at least 97% by weight of the atropisomer (2A) or a salt or tautomer thereof and 0-3% by weight of the atropisomer of formula (2B) or a salt or tautomer thereof.

[0348] 1.172 A composition of matter according to Embodiment 1.168 consisting of at least 98% by weight of the atropisomer (2A) or a salt or tautomer thereof and 0-2% by weight of the atropisomer of formula (2B) or a salt or tautomer thereof.

[0349] 1.173 A composition of matter according to Embodiment 1.168 consisting of at least 99% by weight of the atropisomer (2A) or a salt or tautomer thereof and 0-1% by weight of the atropisomer of formula (2B) or a salt or tautomer thereof.

[0350] 1.174 A composition of matter according to Embodiment 1.168 consisting of at least 99.5% by weight of the atropisomer (2A) or a salt or tautomer thereof and 0-0.5% by weight of the atropisomer of formula (2B) or a salt or tautomer thereof.

[0351] 1.175 A composition of matter according to Embodiment 1.168 consisting of at least 95% by weight of the atropisomer (2B) or a salt or tautomer thereof and 0-5% by weight of the atropisomer of formula (2A) or a salt or tautomer thereof.

[0352] 1.176 A composition of matter according to Embodiment 1.168 consisting of at least 96% by weight of the atropisomer (2B) or a salt or tautomer thereof and 0-4% by weight of the atropisomer of formula (2A) or a salt or tautomer thereof.

[0353] 1.177 A composition of matter according to Embodiment 1.168 consisting of at least 97% by weight of the atropisomer (2B) or a salt or tautomer thereof and 0-3% by weight of the atropisomer of formula (2A) or a salt or tautomer thereof.

[0354] 1.178 A composition of matter according to Embodiment 1.168 consisting of at least 98% by weight of the atropisomer (2B) or a salt or tautomer thereof and 0-2% by weight of the atropisomer of formula (2A) or a salt or tautomer thereof.

[0355] 1.179 A composition of matter according to Embodiment 1.168 consisting of at least 99% by weight of the atropisomer (2B) or a salt or tautomer thereof and 0-1% by weight of the atropisomer of formula (2A) or a salt or tautomer thereof.

[0356] 1.180 The composition of matter of any one of Embodiments 1.168 to 1.179, wherein:

[0357] R 1 Selected from trifluoromethyl, hydroxy, amino, (dimethylamino)methyl and (methoxy)methyl;

[0358] R 2 It is hydrogen;

[0359] R 3 It is hydrogen;

[0360] R 4 Absent or selected from chlorine, fluorine and C 1-4 alkyl;

[0361] Ar 1 is optionally substituted by one or two selected from bromine, fluorine, chlorine, phenoxy, C 1-4 Alkyl (e.g. isopropyl), C 1-4 Alkylsulfonyl (e.g. methylsulfonyl) and cyano substituents R 5 substituted phenyl or pyridyl;

[0362] X is selected from phenyl and pyridyl;

[0363] m is 0 or 1;

[0364] Y is selected from phenyl, pyridyl and thienyl;

[0365] n is 0 or 1;

[0366] R 6 is selected from groups A to AM in Table 1 above; and

[0367] R 7 Selected from chlorine, fluorine and C 1-4 Alkoxy (eg, methoxy).

[0368] 1.181 A composition of matter according to Embodiment 1.180 wherein:

[0369] R 1 is trifluoromethyl;

[0370] R 2 It is hydrogen;

[0371] R 3 It is hydrogen;

[0372] Ar 1 is a substituent R selected from fluorine, chlorine and cyano 5 Substituted phenyl;

[0373] X is phenyl;

[0374] m is 0;

[0375] Y is phenyl or pyridyl;

[0376] n is 0; and

[0377] R 6 is a group (A):

[0378]

[0379] 1.182 A composition of matter according to any one of Embodiments 1.1 and 1.154 to 1.179 wherein the atropisomer is an atropisomer of a compound of any one of Examples A-1 to A-8 and B-2 to B-107.

[0380] 1.183 A composition of matter according to any one of Embodiments 1.1 and 1.154 to 1.179 wherein the atropisomer is an atropisomer of a compound of any one of Examples A-1 to A-8.

[0381] 1.184 A composition of matter consisting of 99.5-100% by weight of a single atropisomer as defined in any one of Embodiments 1.1 to 1.183.

[0382] 1.185 A composition of matter consisting of 99.9-100% by weight of a single atropisomer as defined in any one of Embodiments 1.1 to 1.183.

[0383] 1.186 A single atropisomer having a chemical structure as defined in any one of Embodiments 1.1 to 1.183, said single atropisomer being unaccompanied by any other atropisomer or being accompanied by no more than 0.5% by weight of any other atropisomer relative to said single atropisomer.

[0384] 1.187 A single atropisomer having a chemical structure as defined in any one of Embodiments 1.1 to 1.183, said single atropisomer being unaccompanied by any other atropisomer or being accompanied by no more than 0.25% by weight of any other atropisomer relative to said single atropisomer.

[0385] 1.188 A single atropisomer having a chemical structure as defined in any one of Embodiments 1.1 to 1.183, said single atropisomer being unaccompanied by any other atropisomer or being accompanied by no more than 0.1% by weight of any other atropisomer relative to said single atropisomer.

[0386] 1.188A A single atropisomer according to Embodiment 1.188 having the R configuration represented by formula (1), or a salt thereof:

[0387]

[0388] 1.189 A composition of matter as defined in any one of Embodiments 1.1 to 1.185 or a single atropisomer as defined in any one of Embodiments 1.186 to 1.188A wherein each atropisomer is in the form of a salt.

[0389] 1.190 A composition of matter as defined in any one of Embodiments 1.1 to 1.187 or a single atropisomer as defined in Embodiment 1.188 or 1.188A wherein each atropisomer is in the form of an acid addition salt.

[0390] 1.191 A composition of matter as defined in any one of Embodiments 1.1 to 1.187 or a single atropisomer as defined in Embodiment 1.188 or 1.188A wherein each atropisomer is in non-salt form.

[0391] 1.192 A composition of matter as defined in any one of Embodiments 1.1 to 1.187 or a single atropisomer as defined in Embodiment 1.188 or 1.188A, wherein each atropisomer is in the form of an acid addition salt (preferably having an approximately 1:1 salt ratio) formed with an acid selected from hydrochloric acid, methanesulfonic acid, maleic acid, malic acid, tartaric acid, p-toluenesulfonic acid, phosphoric acid and sulfuric acid.

[0392] A preferred acid addition salt of the present invention is a 1:1 salt formed between a single atropisomer compound (1) as described in Embodiment 1.88A and (+)-L-tartaric acid.

[0393] (+)-L-tartaric acid is particularly advantageous in that it is a highly crystalline and stable solid that absorbs only surface moisture (<1% at 90% RH) with improved water solubility over the free base. These properties make it particularly suitable for drug development.

[0394] Therefore, in further embodiments (Embodiments 1.193 to 1.211), the invention provides:

[0395] 1.193 (R)-4-[5-(4-Chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]benzamide (+)-L-tartrate salt having approximately a 1:1 molar ratio between acid and base.

[0396] 1.194 (+)-L-tartrate salt of 4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]benzamide of formula (2):

[0397]

[0398] 1.195 The (+)-L-tartrate salt of 4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]benzamide, wherein there is approximately a 1:1 molar ratio between the acid and the base and wherein the 4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]benzamide is in the form of a single atropisomer.

[0399] 1.196 A (+)-L-tartrate salt according to Embodiment 1.195 wherein the single atropisomer is an atropisomer of formula (1 ) as defined in Embodiment 1.188A.

[0400] 1.197 A (+)-L-tartrate salt according to Embodiment 1.195 wherein the single atropisomer is the R atropisomer of 4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]benzamide.

[0401] 1.198 A (+)-L-tartrate salt according to Embodiment 1.95, wherein the single atropisomer is characterized by any one or more of the following parameters:

[0402] (i) X-ray crystallographic data substantially as described in Example 3 herein;

[0403] (ii) a retention time of about 20 minutes (e.g., about 20.5 minutes) when determined by Chiral HPLC Method 1 herein; and

[0404] (iii) a specific optical rotation of approximately -11.76° when measured using the method described in Example 2 herein.

[0405] 1.199 A (+)-L-tartrate salt according to Embodiment 1.195 wherein the single atropisomer is atropisomer A-2 as described in the Examples herein.

[0406] 1.200 A (+)-L-tartrate salt according to Embodiment 1.195 wherein the (+)-L-tartrate salt is as described in the Examples herein.

[0407] 1.201 A (+)-L-tartrate salt according to any one of Embodiments 1.193 to 1.200 which is in crystalline form.

[0408] 1.202 A (+)-L-tartrate salt according to Embodiment 1.201 which is an anhydrate.

[0409] 1.203 A (+)-L-tartrate salt according to Embodiment 1.202 which is the anhydrate identified herein as Pattern B.

[0410] 1.204 A (+)-L-tartrate salt according to Embodiment 1.201 which is a solvate.

[0411] 1.205 A (+)-L-tartrate salt according to Embodiment 1.204 which is a solvate identified herein as Pattern A.

[0412] 1.206 A composition of matter comprising a (+)-L-tartrate salt according to any one of embodiments 1.193 to 1.205, wherein (a) the single atropisomer is the only atropisomer of 4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]benzamide present in the composition or (b) less than 10% by mole of any other atropisomer of 4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]benzamide is present relative to the single atropisomer.

[0413] 1.207 A composition of matter according to embodiment 1.206, wherein (a) the single atropisomer is the only atropisomer of 4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]benzamide present in the composition or (b) any other atropisomer of 4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]benzamide is present at less than 5% by mole relative to the single atropisomer.

[0414] 1.208 A composition of matter according to embodiment 1.206, wherein (a) the single atropisomer is the only atropisomer of 4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]benzamide present in the composition or (b) any other atropisomer of 4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]benzamide is present at less than 2% by mole relative to the single atropisomer.

[0415] 1.209 A composition of matter according to embodiment 1.206, wherein (a) the single atropisomer is the only atropisomer of 4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]benzamide present in the composition or (b) any other atropisomer of 4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]benzamide is present at less than 1.5% by mole relative to the single atropisomer.

[0416] 1.210 A composition of matter according to embodiment 1.206, wherein (a) the single atropisomer is the only atropisomer of 4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]benzamide present in the composition or (b) any other atropisomer of 4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]benzamide is present at less than 1% by mole relative to the single atropisomer.

[0417] 1.211 A composition of matter according to embodiment 1.206, wherein (a) the single atropisomer is the only atropisomer of 4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]benzamide present in the composition or (b) any other atropisomer of 4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]benzamide is present at less than 0.1% by mole relative to the single atropisomer.

[0418] definition

[0419] The terms "one or more atropisomer compounds", "one or more atropisomer compounds of the invention", "one or more compounds of formula (1)", "one or more compounds" and "one or more compounds of the invention" may be used herein to refer to the compositions of matter and atropisomers as defined in any of Embodiments 1.1 to 1.211. Unless the context indicates otherwise, such terms may be considered to refer to any of the atropisomers of Formula (1A), (1B), (2A) and (2B) and all subgroups, preferences, embodiments and examples as defined herein. The term "compound of formula (1)" may be used herein as a general term to cover atropisomers of Formula (1A), (1B), (2A) and (2B) and all subgroups, preferences, embodiments and examples thereof and mixtures of atropisomers. It will be apparent from the context that where a compound of formula (1) is referred to, it is not necessarily the case whether it refers to an individual atropisomer, a composition of matter or a mixture of atropisomers.

[0420] As used herein, the term 'drug' refers to a pharmaceutical preparation used to treat, cure or ameliorate a disease or to treat, alleviate or relieve symptoms of a disease. A pharmaceutical preparation includes a pharmacologically active ingredient in a form that is harmless to the subject to which it is administered and additional components designed to stabilize the active ingredient and affect its absorption into the circulation or target tissues.

[0421] Salt

[0422] Where atropisomers as defined in any one of Embodiments 1.1 to 1.188A contain ionizable groups, they may be present in the form of salts as defined in any one of Embodiments 1.189, 1.190 and 1.192 to 1.211.

[0423] For example, where the atropisomer contains a basic (eg, nitrogen basic) group or atom, the atropisomer may be present as an acid addition salt.

[0424] Salts can be synthesized from parent compounds by conventional chemical methods, such as those described in Pharmaceutical Salts: Properties, Selection, and Use, P. Heinrich Stahl (Editor), Camille G. Wermuth (Editor), ISBN: 3-90639-026-8, Hardcover, 388 pages, August 2002. In general, such salts can be prepared by reacting the free base form of the compound with an acid in water or in an organic solvent, or in a mixture of the two; generally, a non-aqueous medium such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile is used.

[0425] Acid addition salts (as defined in Embodiment 1.190) can be formed with a wide variety of acids, both inorganic and organic. Examples of acid addition salts include salts formed with an acid selected from the group consisting of acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid (e.g., L-ascorbic acid), L-aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, butyric acid, (+) camphoric acid, camphorsulfonic acid, (+)-(1S)-camphor-10-sulfonic acid, decanoic acid, hexanoic acid, caprylic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, mucic acid, gentisic acid, glucoheptonic acid, D-gluconic acid, glucuronic acid (e.g., D-glucuronic acid), glutamic acid (e.g., L-glutamic acid), α-oxygenase Glutaric acid, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, hydroiodic acid, isethionic acid, (+)-L-lactic acid, (±)-DL-lactic acid, lactobionic acid, maleic acid, malic acid, (-)-L-malic acid, malonic acid, (±)-DL-mandelic acid, methanesulfonic acid, naphthalene-2-sulfonic acid, naphthalene-1,5-disulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, nitric acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, phosphoric acid, propionic acid, L-pyroglutamic acid, salicylic acid, 4-amino-salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, tannic acid, (+)-L-tartaric acid, thiocyanic acid, p-toluenesulfonic acid, undecylenic acid and valeric acid, as well as acylated amino acids and cation exchange resins.

[0426] The salt forms of the compositions of matter or atropisomers of the invention are generally pharmaceutically acceptable salts, and examples of pharmaceutically acceptable salts are discussed in Berge et al., 1977, "Pharmaceutically Acceptable Salts," J. Pharm. Sci., Vol. 66, pp. 1-19. However, non-pharmaceutically acceptable salts may also be prepared as intermediate forms, which may then be converted into pharmaceutically acceptable salts. Such non-pharmaceutically acceptable salt forms may be used, for example, to purify or isolate the compositions of matter or atropisomers of the invention, and also form part of the invention.

[0427] Geometric isomers and tautomers

[0428] In addition to existing as atropisomers, the compositions of matter or atropisomers of the invention may also contain other structural features that give rise to geometric isomerism and tautomerism, and references to compositions of matter or atropisomers as defined in Embodiments 1.1 to 1.211 include all geometric isomers and tautomeric forms. For the avoidance of doubt, where an atropisomer may exist in one of several geometric isomers or tautomeric forms and only one is specifically described or shown, all others are nonetheless encompassed by Formula (1A) (1B) or subgroups, subsets, preferences and examples thereof.

[0429] Optical isomers

[0430] Where the compounds of the invention contain one or more chiral centers in addition to structural features that give rise to atropisomerism, reference to a composition of matter or an atropisomer includes all optical isomeric forms thereof (e.g., enantiomers, epimers, and diastereomers), either as individual optical isomers or as mixtures thereof (other than mixtures of atropisomers), unless the context requires otherwise.

[0431] Optical isomers can be characterized and identified by their optical activity (i.e., + and - isomers, or d and l isomers), or they can be characterized by their absolute stereochemistry using the "R and S" nomenclature developed by Cahn, Ingold, and Prelog, see Advanced Organic Chemistry by Jerry March, 4 th Edition, John Wiley & Sons, New York, 1992, pages 109-114, and see also Cahn, Ingold & Prelog, Angew. Chem. Int. Ed. Engl., 1966, 5, 385-415.

[0432] Optical isomers may be separated by a number of techniques including chiral chromatography (chromatography on a chiral support), and such techniques are well known to those skilled in the art.

[0433] As an alternative to chiral chromatography, the optical isomers can be separated by forming diastereomeric salts with chiral acids such as (+)-tartaric acid, (-)-pyroglutamic acid, (-)-di-toluoyl-L-tartaric acid, (+)-mandelic acid, (-)-malic acid, and (-)-camphorsulfonic acid, separating the diastereomers by preferential crystallization, and then dissociating the salts to give the individual enantiomers of the free base.

[0434] When the compounds of the present invention exist in two or more optical isomeric forms, one enantiomer of a pair of enantiomers may exhibit advantages over the other enantiomer, for example, in terms of biological activity. Therefore, in some cases, it may be desirable to use only one of a pair of enantiomers, or only one of multiple diastereomers as a therapeutic agent. Therefore, the present invention provides a composition comprising an atropisomer having one or more chiral centers, wherein at least 55% (e.g., at least 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95%) of the composition of matter or the atropisomer of formula (1) exists as a single optical isomer (e.g., enantiomer or diastereomer). In a general embodiment, 99% or more (e.g., substantially all) of the total amount of the composition of matter or the atropisomer of formula (1) may exist as a single optical isomer (e.g., enantiomer or diastereomer).

[0435] isotope

[0436] A composition of matter or atropisomer of the invention as defined in any one of Embodiments 1.1 to 1.211 may contain one or more isotopic substitutions, and reference to a specific element includes within its scope all isotopes of that element. For example, reference to hydrogen includes within its scope all isotopes of hydrogen. 1 H. 2 H(D) and 3 H(T). Similarly, references to carbon and oxygen include within their respective scopes 12 C. 13 C and 14 C and 16 O and 18 O.

[0437] Isotopes can be radioactive or non-radioactive. In one embodiment of the invention, the composition of matter or atropisomer does not contain a radioactive isotope. Such compounds are preferably used for therapeutic purposes. In another embodiment, however, the composition of matter or atropisomer may contain one or more radioactive isotopes. Compounds containing such radioactive isotopes may be useful in a diagnostic context.

[0438] Solvate

[0439] A composition of matter or atropisomer as defined in any one of Embodiments 1.1 to 1.211 may form solvates and anhydrates.

[0440] A specific solvate is a solvate formed by incorporating molecules of a non-toxic pharmaceutically acceptable solvent (hereinafter referred to as a solvating solvent) into the solid state structure (e.g., crystal structure) of the composition of matter of the present invention or atropisomer. Examples of such solvents include water, alcohols (such as ethanol, isopropanol, and butanol) and dimethyl sulfoxide. Solvates can be prepared by recrystallizing the composition of matter or atropisomer of the present invention with a solvent or solvent mixture containing a solvating solvent. In any given case, whether a solvate has been formed can be determined by subjecting the crystals of the composition of matter or atropisomer to analysis using well-known and standard techniques such as thermogravimetric analysis (TGE), differential scanning calorimetry (DSC), and X-ray powder diffraction (XRPD).

[0441] Solvates may be stoichiometric or non-stoichiometric solvates.

[0442] A particularly preferred solvate is a hydrate, and examples of the hydrate include a hemihydrate, a monohydrate, and a dihydrate.

[0443] For a more detailed discussion of solvates and methods for making and characterizing them, see Bryn et al., Solid-State Chemistry of Drugs, Second Edition, published by SSCI, Inc of West Lafayette, IN, USA, 1999, ISBN 0-967-06710-3.

[0444] In addition to forming solvates, the composition of matter, compound or salt as defined in any one of Embodiments 1.1 to 1.211 may also be provided in an anhydrate form. The term "anhydrate" as used herein refers to a solid particulate form (e.g., a crystalline form) that does not contain water (and preferably does not contain any other solvent) within its three-dimensional structure, although the particles of the salt or compound may have water molecules attached to their outer surface.

[0445] Prodrug

[0446] A compound, salt, composition of matter or atropisomer as defined in any one of Embodiments 1.1 to 1.211 may be presented in the form of a prodrug.

[0447] By "prodrug" is meant any compound that is converted in vivo to a biologically active composition of matter or atropisomer as defined in any one of Embodiments 1.1 to 1.211, for example.

[0448] For example, some prodrugs are esters of the active compound (e.g., physiologically acceptable metabolically labile esters). During metabolism, the ester group (-C(=O)OR) is cleaved to produce the active drug. Such esters can be formed by esterification of, for example, any hydroxyl groups present in the parent compound, if necessary, where appropriate, pre-protecting any other reactive groups present in the parent compound, and then deprotecting.

[0449] In addition, some prodrugs are enzymatically activated to produce the active compound, or to produce a compound that produces the active compound after another chemical reaction (e.g., such as ADEPT, GDEPT, LIDEPT, etc.). For example, the prodrug may be a sugar derivative or other glycoside conjugate, or may be an amino acid ester derivative.

[0450] Process for preparing the compounds of the present invention

[0451] Compositions of matter and atropisomers of the present invention can be prepared by separating mixtures of atropisomers using chiral chromatography, and in particular chiral HPLC.

[0452] Mixtures of atropisomers of formula (1A), (1B), (2A), (2B) and various subgroups thereof can be prepared according to synthetic methods well known to the skilled artisan. 1 –R 7 ,Ar 1 , X, Y and Z are as defined above. In the following paragraphs relating to the preparation of a mixture of atropisomers, the mixture will generally be referred to as a compound of formula (1).

[0453] Compounds of formula (1) wherein Z is a pyrrole ring can be prepared by reacting a 1,4-dicarbonyl compound of formula (10) with an aminoaryl compound of formula (11), as shown in Scheme 1.

[0454]

[0455] The starting materials for the synthetic route shown in Scheme 1 are 1-aryl-3-bromoacetone (12) and arylacetone (13), both of which are commercially available.

[0456] 1-Aryl-2-bromoethanone (12) is reacted with arylacetone (13) to produce a 1,4-dicarbonyl compound (10). The reaction is preferably carried out in a non-polar, aprotic solvent (e.g., benzene or toluene) in the presence of a zinc (II) salt (e.g., zinc chloride). Preferably, a tertiary alcohol (e.g., tert-butyl alcohol) and a tertiary amine (e.g., triethylamine) are also added. The reaction can be carried out at room temperature for a period of, for example, 12 to 48 hours.

[0457] The 1,4-dicarbonyl compound (10) can then be reacted with an aminoarene (11) to form the trisubstituted pyrrole compound (1) of the present invention. The reaction can be carried out in a non-polar, aprotic solvent (e.g., dioxane). The reaction mixture can be heated (e.g., between 150 and 170° C.) and / or microwave irradiated. The reaction can be carried out for between 1 and 12 hours, such as between 1 and 6 hours. A strong acid (e.g., p-toluenesulfonic acid) can also be added as a catalyst.

[0458] Alternatively, the synthetic route shown in Scheme 2 can be used to prepare 2 and R 3 The compound of formula (10) wherein all of them are hydrogen.

[0459]

[0460] The starting aldehyde (11a) can be prepared by reducing the corresponding acid with a reducing agent such as NaBH4, followed by oxidation with a suitable oxidizing agent. Such an example of an oxidizing agent for preparing an aldehyde without further oxidation to a carboxylic acid is Dess-Martin periodinane. The starting amine (11b) can be prepared by a Mannich reaction of dimethylamine hydrochloride and formaldehyde in the presence of an acid catalyst in a polar, protic solvent such as ethanol.

[0461] Compounds of formula (10) can then be prepared by reacting compounds (11a) and (11b) in a polar, aprotic solvent (e.g., 1,2-dimethoxyethane) with a suitable catalyst. One such suitable catalyst is a thiazolium salt (e.g., 3-ethyl-5-(2-hydroxyethyl)-4-methylthiazolium bromide). The reaction is typically carried out at an elevated temperature (e.g., between 80°C and 120°C) for between 1 and 24 hours, even more preferably between 2 and 12 hours.

[0462] Once formed, one compound of formula (1) can be converted to another compound of formula (1) using standard chemical procedures well known in the art. For examples of functional group interchange, see, for example, March's Advanced Organic Chemistry, Michael B. Smith & Jerry March, 6 th Edition, Wiley-Blackwell (ISBN: 0-471-72091-7), 2007 and Organic Syntheses, Volumes 1-9, John Wiley, edited by Jeremiah P. Freeman (ISBN: 0-471-12429), 1996.

[0463] The compounds of formula (1) can be prepared by following the synthetic route as shown in Scheme 3, wherein Y is replaced by a substituent R 6 Substitution, where R 6 The formula is C(O)NHR 8 An amide group, wherein R 8 is an optionally substituted C 1-8 Hydrocarbon group.

[0464]

[0465] In Scheme 3, Y represents Ring Y as defined herein.

[0466] The compound of formula (14) can be prepared according to the synthetic route shown in Scheme 1 above, wherein R 11 It is C 1-8 Hydrocarbyl groups or other carboxylic acid protecting groups. The ester (14) can be hydrolyzed to obtain the carboxylic acid (15). This is preferably carried out in a mixture of a non-polar, aprotic solvent (e.g., tetrahydrofuran) and a polar, protic solvent (e.g., water). One such suitable solvent system is a 1:1 mixture of tetrahydrofuran and water. A water-soluble strong base (e.g., lithium hydroxide) is added and the reaction mixture is stirred at room temperature for an extended period of time, e.g., between 6 and 48 hours, more typically between 12 and 48 hours.

[0467] The acid compound (15) is then reacted with the corresponding amine (H2N-R) under amide forming conditions, for example in the presence of a reagent of the type commonly used to form amide bonds. 8 ) to provide a compound of formula (1), wherein R 6 is an amide. Examples of such reagents include carbodiimide coupling agents, such as 1,3-dicyclohexylcarbodiimide (DCC) (Sheehan et al, J. Amer. Chem Soc. 1955, 77 , 1067) and 1-ethyl-3-(3'-dimethylaminopropyl)-carbodiimide (herein referred to as EDC or EDCI) (Sheehan et al, J. Org. Chem., 1961, 26, 2525), which is usually combined with 1-hydroxy-7-azabenzotriazole (HOAt) (LA Carpino, J. Amer. Chem. Soc., 1993, 115, 4397) or 1-hydroxybenzotriazole (HOBt) (Konig et al, Chem. Ber., 103, 708, 2024-2034). Another example of such a reagent is a uronium coupling agent, such as O-(7-azabenzotriazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU). A preferred amide coupling agent is HATU.

[0468] The coupling reaction is typically performed in a non-aqueous, aprotic solvent such as dimethylformamide at room temperature in the presence of a non-interfering base (eg, a tertiary amine such as triethylamine or N,N-diisopropylethylamine).

[0469] Alternatively, suitable hydrolysis conditions can be used to prepare compounds of formula (15) by the hydrolysis of corresponding nitrile. Preferably, a strong base is used in a polar protic solvent or a mixture of polar protic solvents, such as an alkali metal hydroxide (e.g., sodium hydroxide) to be hydrolyzed. An example of such a suitable solvent system is a mixture of methanol and water. The reaction is preferably carried out between 12 and 24 hours at an elevated temperature.

[0470] The compounds of formula (1) can be prepared according to the synthetic route shown in Scheme 4, wherein Y is replaced by a substituent R 6 Substitution, where R 6 Is a NHR 9 of amine groups.

[0471]

[0472] In Scheme 4, Y represents Ring Y as defined herein.

[0473] Compounds of formula (16) can be prepared according to the synthetic route shown in Scheme 1 above. Compound (16) can then be reduced to compound (17) using a suitable reducing agent (e.g., sodium borohydride) and optionally a catalytic amount of a copper (II) salt (e.g., copper (II) acetate). The reaction is preferably carried out in an anhydrous, polar, aprotic solvent (e.g., methanol).

[0474] Then compound (17) can be reacted with the formula LG-R 9 wherein LG is a suitable leaving group (e.g., halogen, more preferably chlorine), and R 9 is an optionally substituted non-aromatic C 1-8 The amine compound (17) is first treated with a suitable base (e.g., sodium hydride) in a polar, aprotic solvent (e.g., dimethylformamide), typically at room temperature, and then reacted with the compound LG-R 9 , typically reacting at elevated temperature (eg, between 60°C and 100°C).

[0475] Alternatively, compounds of formula (1) can be prepared from compounds of formula (17) and carboxylic acids or activated derivatives such as acid chlorides or anhydrides in a manner analogous to that shown in Scheme 4, wherein R 6 It is an amide in which the nitrogen atom of the amide is bonded to ring Y.

[0476] Alternatively, according to Scheme 5, compounds of formula (1) can be prepared from intermediates (17) under amide forming conditions, for example in the presence of reagents of the type commonly used to form amide bonds, wherein R 6 It is NHCOR 10 An amide, wherein R 10 is an optionally substituted C 1-8 Hydrocarbon group.

[0477]

[0478] In Scheme 5, Y represents Ring Y as defined herein.

[0479] Examples of such reagents include carbodiimide coupling agents such as 1,3-dicyclohexylcarbodiimide (DCC) (Sheehan et al, J. Amer. Chem Soc. 1955, 77 , 1067) and 1-ethyl-3-(3'-dimethylaminopropyl)-carbodiimide (herein referred to as EDC or EDCI) (Sheehan et al, J. Org. Chem., 1961, 26, 2525), which is usually combined with 1-hydroxy-7-azabenzotriazole (HOAt) (LA Carpino, J. Amer. Chem. Soc., 1993, 115 , 4397) or 1-hydroxybenzotriazole (HOBt) (Konig et al, Chem. Ber., 103, 708, 2024-2034). Another example of such a reagent is a uronium coupling agent, such as O-(7-azabenzotriazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU). A preferred amide coupling agent is HATU.

[0480] The coupling reaction is typically performed in a non-aqueous, aprotic solvent such as dimethylformamide at room temperature in the presence of a non-interfering base (eg, a tertiary amine such as triethylamine or N,N-diisopropylethylamine).

[0481] The compounds of formula (1) can be prepared by following the synthetic route as shown in Scheme 6, wherein Y is replaced by a substituent R 6 Substitution, where R 6 Is a formula OR 12 An ether group, wherein R12 is an optionally substituted C 1-8 Hydrocarbon group.

[0482]

[0483] In Scheme 6, Y represents Ring Y as defined herein.

[0484] Compounds of formula (19) can be prepared according to the synthetic route shown in Scheme 1 above. Compound (19) can then be reacted with a compound of formula LG-R 12 wherein LG is a suitable leaving group (e.g., halogen, more preferably chlorine), and R 7 is an optionally substituted non-aromatic C 1-8 The alcohol compound (19) is first deprotonated with a suitable base (e.g., sodium hydride) in a polar, aprotic solvent (e.g., dimethylformamide). The reaction can be carried out at room temperature. 12 The second step of the reaction may be carried out at elevated temperature, typically between 80°C and 100°C.

[0485] The compounds of formula (1) can be prepared according to Scheme 7, wherein R 6 It's Q 1 -R a -R b And Q 1 It is a methylene group.

[0486]

[0487] In Scheme 7, Y represents Ring Y as defined herein.

[0488] Compound (15) (available as described in Scheme 3 above) is treated with a reducing agent (e.g., sodium borohydride) in a polar aprotic solvent such as tetrahydrofuran to provide the primary alcohol (20). The alcohol (20) can then be reacted as described in Scheme 6 above to provide a compound wherein R 6 Another compound of formula (1) is an ether.

[0489] Alternatively, compound (20) can undergo other standard functional group interchanges to produce additional compounds of formula (1), such as by oxidation to aldehydes and by reductive amination to form amines. The amines produced by this method can be further reacted with carboxylic acids or acid derivatives to produce amide compounds of formula (1) using the methods described in Scheme 5 above.

[0490] Compounds of formula (1) wherein Z is a 1,4,5-trisubstituted pyrazole can be prepared by reacting an aryl hydrazine (21) with an α,β-unsaturated carbonyl compound (22), as shown in Scheme 8.

[0491]

[0492] In Scheme 8, X and Y represent rings X and Y, respectively, as defined herein.

[0493] The aryl hydrazine (21) and the α,β-unsaturated carbonyl compound (22) are dissolved in a suitable polar, protic solvent system (e.g., 1:1 water:methanol) with a suitable base (e.g., sodium carbonate). The mixture is typically stirred at or about room temperature (e.g., for about 15 minutes) before a weak acid such as acetic acid is added. The resulting mixture is then heated (e.g., between 100° C. and 140° C. for an extended period of time (e.g., between 6 and 12 hours) for a sufficient period of time (e.g., 8 hours)) to provide a compound of formula (1), wherein Z is a 1,4,5-trisubstituted pyrazole.

[0494]

[0495] The starting α,β-unsaturated carbonyl compound (22) of Scheme 8 can be prepared from the corresponding ketone (23) and N,N-dimethylformamide dimethyl acetal. A solution of N,N-dimethylformamide dimethyl acetal in a polar aprotic solvent such as DMF is added to a solution of ketone (23). The mixture is typically heated to a temperature between, for example, 70° C. and 110° C. (e.g., about 90° C.) to provide compound (22). Compound (23) can be prepared by Ar 1 Grignard reaction between CH2CHO and Br-X is then obtained by oxidation of the resulting alcohol with a suitable oxidant (eg, Dess-Martin periodinane) in a solvent such as DCM to provide the ketone (23).

[0496] Alternatively, when alternative isomers of formula (1) wherein Z is a 3,4,5-trisubstituted pyrazole are desired, these isomers may be prepared as described in Scheme 10 below.

[0497]

[0498]

[0499] In Scheme 10, X and Y represent rings X and Y, respectively, as defined herein.

[0500] The alkenyl bromide (25) is reacted with the diazo compound (26) in a 1,3-dipolar cycloaddition reaction by mixing the two compounds with a strong base (eg, sodium hydroxide) and heating (eg, to a temperature of about 70°C) to provide the bromopyrazole (27).

[0501] The bromopyrazole (27) is then reacted with an organoboronic acid having the formula XB(OH)2 (wherein X is a ring as defined herein) in the presence of a palladium(0) catalyst, such as bis(tri-tert-butylphosphine)palladium(0) and a suitable base, such as a carbonate or phosphate of cesium or potassium, under Suzuki reaction conditions in a polar solvent such as dioxane to obtain a compound of formula (1) wherein Z is pyrazole or a protected derivative thereof. The bromopyrazole (27) may be in a protected form. For example, in the NH group on the pyrazole, a protecting group such as a Boc (tert-butyloxycarbonyl) group may be attached to the nitrogen atom, replacing the hydrogen atom. After the reaction between the organoboronic acid and the pyrazole (27), a deprotection step may be required to obtain a compound of formula (1). In the case of the Boc protecting group, this may be removed by treatment with an acid, such as hydrochloric acid.

[0502] Boronic acids and organic boronic acids are widely available commercially or can be prepared, for example, as described in the review article by N. Miyaura and A. Suzuki, Chem. Rev. 1995, 95, 2457. Thus, organic boronic acids can be prepared by reacting the corresponding bromine compound with an alkyl lithium such as butyl lithium and then with a borate ester. If desired, the resulting organic boronic acid derivative can be hydrolyzed to obtain the corresponding organic boronic acid.

[0503] Starting material (25) can be prepared by treating an aryl aldehyde with carbon tetrabromide and triphenylphosphine in a solvent such as DCM at low temperature (e.g., about 0° C.). Starting material (26) can be prepared from the corresponding aryl aldehyde by treating with p-toluenesulfonylhydrazide in a polar protic solvent such as methanol and heating (e.g., to about 60° C.).

[0504] Compounds of formula (1) wherein Z is an isoxazole group can be prepared according to the synthetic scheme in Scheme 11.

[0505]

[0506] In Scheme 11, X and Y represent rings X and Y, respectively, as defined herein.

[0507] Intermediate (30) can be prepared by reacting alkyne (28) with oxime (29) in a polar, aprotic solvent such as diethyl ether by mixing with a base such as triethylamine, for example at a temperature near room temperature, to provide isoxazole (30). Isoxazole (30) can then be brominated with a suitable brominating agent such as N-bromosuccinimide as a bromine source to provide bromoisoxazole (31). The reaction is typically carried out at elevated temperatures (e.g., between 90° C. and 120° C.) in an acidic solution (e.g., acetic acid).

[0508] Then, in the presence of a palladium (0) catalyst, such as bis (tri-tert-butylphosphine) palladium (0) and a base (e.g., carbonate or phosphate of cesium or potassium), under Suzuki reaction conditions, in a polar solvent such as dioxane, bromoisoxazole (31) is reacted with an organic boronic acid having the formula XB (OH) 2 (wherein X is a ring as defined herein) to obtain a compound of formula (1), wherein Z is isoxazole or a protected derivative thereof. Bromoisoxazole (31) may be in a protected form. For example, in the NH group on the group Ar1 or Y, a protecting group such as a Boc (tert-butyloxycarbonyl) group may be attached to the nitrogen atom to replace the hydrogen atom. After the reaction between the organic boronic acid and isoxazole (31), a deprotection step may be required to obtain a compound of formula (1). In the case of the Boc protecting group, this can be removed by treatment with an acid (such as hydrochloric acid).

[0509] Boronic acids and organic boronic acids are widely available commercially or can be prepared, for example, as described in the review article by N. Miyaura and A. Suzuki, Chem. Rev. 1995, 95, 2457. Thus, organic boronic acids can be prepared by reacting the corresponding bromine compound with an alkyl lithium such as butyl lithium and then with a borate ester. If desired, the resulting organic boronic acid derivative can be hydrolyzed to obtain the corresponding organic boronic acid.

[0510] The starting material (29) can be prepared from the corresponding aryl aldehyde by a two-step process. The first step consists of treating the aldehyde with NH2OH and a strong base such as sodium hydroxide in a polar protic solvent system such as 1:1 ethanol:water to provide the aryl oxime. This can then be chlorinated by mixing it with N-chlorosuccinimide in dimethylformamide and stirring for 18 hours to provide the starting material (29).

[0511] The synthesis of compounds of formula (1) has been illustrated above using reaction schemes for the preparation of pyrroles, isoxazoles and pyrazoles. However, it will be readily appreciated that similar methods may be used to prepare compounds of formula (1) containing other five-membered heteroaryl rings.

[0512] A specific synthetic route for preparing the preferred atropisomer of the present invention, compound (1) is shown in Scheme 12 below.

[0513]

[0514] The starting materials for the synthetic route shown in Scheme 1 are 4-cyanogen-acetophenone (104) and 4-chlorophenacyl bromide (105), both of which are commercially available.

[0515] In step 1, 4-cyano-acetophenone (104) and 4-chlorophenacyl bromide (105) are reacted together to produce 4-[4-(4-chlorophenyl)-4-oxyde-butyryl]benzonitrile (106). The reaction is typically carried out in a suitable solvent, e.g., a mixture of a non-polar (e.g., hydrocarbon) solvent (such as benzene or toluene) and a tertiary alcohol (e.g., tert-butyl alcohol), in the presence of a tertiary amine such as triethylamine, in the presence of a zinc (II) salt (e.g., zinc chloride). The reaction can be carried out at or near room temperature, e.g., for a period of 12 to 60 hours.

[0516] In step 2, 4-[4-(4-chlorophenyl)-4-oxyylidene-butyryl]benzonitrile (106) is reacted with 2-trifluoromethylaniline to produce 4-(5-(4-chlorophenyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrrol-2-yl)benzonitrile (107). The reaction is typically carried out in the presence of an acid catalyst such as p-toluenesulfonic acid in a suitable high boiling point solvent such as dioxane at elevated temperature (e.g., between 130° C. and 170° C.) and / or under microwave irradiation. The reaction may be carried out for between 1 and 12 hours, for example, between 1 and 6 hours.

[0517] In step 3, 4-(5-(4-chlorophenyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrrol-2-yl)benzonitrile (107) is subjected to alkaline hydrolysis to produce 4-(5-(4-chlorophenyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrrol-2-yl)benzoic acid (108). The hydrolysis reaction is typically conducted in an aqueous solvent (which may contain an alcohol such as methanol) in the presence of an alkali metal hydroxide such as sodium hydroxide (typically in excess), and is typically accompanied by heating, for example, to a temperature in the range of 60-80° C. or for a period of up to about 20 hours or more. Once the hydrolysis is complete, the acid (8) is typically isolated by cooling and acidifying the reaction mixture.

[0518] After step 3, one of two possible routes to atropisomers (1) can be followed. In a variation consisting of steps 4b and 5b and 6, 4-(5-(4-chlorophenyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrrol-2-yl)benzoic acid (108) is reacted with N,N-dimethylethylenediamine under amide forming conditions to produce a racemic mixture of atropisomers of 4-(5-(4-chlorophenyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrrol-2-yl)-N-(2-(dimethylamino)ethyl)benzamide (109), which is then resolved into its individual atropisomers by chiral separation to produce atropisomers (1).

[0519] In another variation, racemic 6(4-(5-(4-chlorophenyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrrol-2-yl)benzoic acid (108)) undergoes chiral separation to produce atropisomer (103), which is then reacted with N,N-dimethylethylenediamine under amide forming conditions to produce atropisomer (1).

[0520] Carboxylic acids (103) and (108) are reacted with N,N-dimethylethylenediamine under amide-forming conditions in the presence of an amide coupling reagent. Examples of such amide coupling reagents include carbodiimide-based coupling reagents such as 1,3-dicyclohexylcarbodiimide (DCC) (Sheehan et al, J. Amer. Chem Soc. 1955, 77 , 1067) and 1-ethyl-3-(3'-dimethylaminopropyl)-carbodiimide (herein referred to as EDC or EDCI) (Sheehan et al, J. Org. Chem., 1961, 26, 2525), which is usually combined with 1-hydroxy-7-azabenzotriazole (HOAt) (LA Carpino, J. Amer. Chem. Soc., 1993, 115 , 4397) or 1-hydroxybenzotriazole (HOBt) (Konig et al, Chem. Ber., 103, 708, 2024-2034), uronium coupling reagents such as O-(7-azabenzotriazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) and propanephosphonic anhydride (T3P) (see A. Garcia, Synlett, 2007, No. 8, pp1328-1329). The specific amide coupling reagents used in method steps 5a and 5b are HATU and T3P.

[0521] The amide coupling reaction is typically performed in a non-aqueous, polar, aprotic solvent such as tetrahydrofuran or dimethylformamide or mixtures thereof at or near room temperature (e.g., 18-30° C.) in the presence of a non-interfering base (e.g., a tertiary amine such as triethylamine or N,N-diisopropylethylamine).

[0522] Certain aspects of the methods described above represent additional embodiments of the invention (embodiments 2.1 to 2.8). Accordingly, the present invention provides:

[0523] 2.1 A method of preparing a composition of matter or a single atropisomer as defined in any one of Embodiments 1.1 to 1.211, the method comprising chiral separation of a mixture of atropisomers of a compound of formula (0):

[0524]

[0525] Among them, ring X, ring Y, ring Z, Ar 1 , m, n and R 1 To R 7 As defined in any one of embodiments 1.1 to 1.211.

[0526] 2.2 The method according to Embodiment 2.1, wherein the mixture of atropisomers of the compound of formula (0) is a racemic mixture.

[0527] 2.3 The method according to embodiment 2.1 or embodiment 2.2, wherein the chiral separation is performed by:

[0528] (i) passing the mixture of atropisomers through a chiral chromatographic column; for example, a chiral HPLC column; or

[0529] (ii) reacting a mixture of atropisomers of the compound of formula (0) with a chiral acid to form salts of two atropisomers in the mixture, separating the salts and decomposing the salts to produce the corresponding free base of each atropisomer.

[0530] 2.4 A process for the preparation of an atropisomer (1) as defined herein, said process comprising reacting a compound of formula (103) with N,N-dimethylethylenediamine under amide forming conditions.

[0531] 2.5 A method according to embodiment 2.4, wherein the conditions for forming an amide include the presence of an amide coupling reagent, for example, an amide coupling reagent as described herein.

[0532] 2.6 A method according to embodiment 2.5, wherein the amide coupling reagent is propanephosphonic anhydride (T3P).

[0533] 2.7 A process for preparing a compound of formula (103) (see Scheme 12), which comprises chirally separating a compound of formula (103) from a mixture of atropisomers of formula (108), for example by chiral chromatography or salification with a chiral base and resolution of the resulting chiral salt.

[0534] 2.8 An atropisomer compound of formula (103) or a salt thereof (eg a metal salt such as an alkali metal salt or an alkaline earth metal salt or a salt with ammonia or an organic amine).

[0535] The atropisomers and compositions of matter of the invention may be provided in salt form or in non-salt (eg, free base) form.

[0536] The acid addition salts of the basic atropisomers of the invention can be prepared by contacting the free base form of the atropisomer with a suitable salt-forming acid in a suitable solvent or solvent mixture as described elsewhere herein and then isolating the desired salt from the solvent or solvent mixture.

[0537] A particular salt of the invention is the (+)-L-tartrate salt of formula (2) as defined in any one of Embodiments 1.194 to 1.211.

[0538] The (+)-L-tartrate salt of the present invention can be prepared from the atropisomer of formula (1) by reaction with tartaric acid in a solvent or solvent mixture and then isolating the tartrate salt from the solvent or solvent mixture.

[0539] In one embodiment (Embodiment 2.9), the atropisomer of formula (1) can be dissolved or suspended in one solvent to form a first mixture, and (+)-L-tartaric acid can be dissolved or suspended in the same or another solvent to form a second mixture, and the first and second mixtures are then combined and set aside (e.g., with stirring) for a period of time to allow salt formation to occur, followed by isolation of the (+)-L-tartrate salt.

[0540] When the first and second mixtures are combined, preferably the molar amounts of the atropisomer of formula (1) and the molar amounts of (+)-L-tartaric acid are approximately equal; that is, there is preferably a 1:1 molar ratio between the atropisomer of formula (1) and (+)-L-tartaric acid.

[0541] The (+)-L-tartrate salt can be isolated from the combined mixture by filtration (when a precipitate forms) or by evaporation of the solvent.

[0542] Thus, when more than one solvent is present in the combined mixture, different solvents may be selected to act as co-solvents or to act as anti-solvents.

[0543] The solvent or solvent mixture may be selected such that they retain the (+)-L-tartrate salt at least partially in solution when heated, but then cause the salt to precipitate as a precipitate upon cooling of the solvent or solvent mixture.

[0544] The solvent used to form the first mixture (a mixture comprising atropisomers of formula (1)) can be, for example, selected from aliphatic ketones, aliphatic esters of aliphatic acids, non-aromatic cyclic ethers, and aliphatic alcohols.

[0545] A specific example of the aliphatic ketone is acetone.

[0546] Examples of aliphatic esters of aliphatic acids include C 2-4 Alkyl esters, a specific example is isopropyl acetate.

[0547] Examples of the non-aromatic cyclic ether include dioxane, 2-methyltetrahydrofuran and tetrahydrofuran, and a specific example is 2-methyltetrahydrofuran.

[0548] Examples of aliphatic alcohols are C 2-4 Aliphatic alcohols, and more particularly C 3-4 Alkanols such as isopropanol and butanol.

[0549] The solvent used to form the second mixture (the mixture containing (+)-L-tartaric acid) may be selected, for example, from water, non-aromatic cyclic ethers, and aliphatic alcohols.

[0550] A specific example of the aliphatic alcohol solvent for the second mixture is ethanol.

[0551] A specific example of the non-aromatic cyclic ether solvent for the second mixture is tetrahydrofuran (THF).

[0552] Another specific example of the solvent used to form the second mixture is water.

[0553] The (+)-L-tartrate salt of the atropisomer of formula (1) can exist in several crystalline forms, in particular pattern A (which is a solvate) and pattern B (which is an anhydrate). Characteristic details of the different crystalline forms are provided elsewhere herein. The different crystalline forms can be prepared by varying the solvent and heating conditions used in forming the salt.

[0554] In a method for making a (+)-L-tartrate salt of an atropisomer of formula (1) having pattern A (embodiment 2.10), a solution of the atropisomer in acetone is mixed with a solution of (+)-L-tartaric acid in ethanol at a temperature in the range of 20°C to 30°C (e.g., approximately 25°C), the resulting mixture is stirred or otherwise agitated for a length of time sufficient to allow salt formation to occur (e.g., 12-24 hours), and the salt is then isolated by filtration.

[0555] In another method for making a (+)-L-tartrate salt of a transisomer of formula (1) having pattern A (embodiment 2.11), a solution of the transisomer in isopropanol is mixed with a solution of (+)-L-tartaric acid in ethanol at a temperature in the range of 35°C to 45°C (e.g., approximately 40°C), the resulting mixture is cooled to a temperature in the range of 20°C to 30°C (e.g., approximately 25°C) over a period of about 1-3 hours, and the salt is then separated by filtration.

[0556] In another method for making a (+)-L-tartrate salt of an atropisomer of formula (1) having pattern A (embodiment 2.12), a solution of the atropisomer in 2-methyltetrahydrofuran is mixed with a solution of (+)-L-tartaric acid in ethanol at a temperature in the range of 20°C to 30°C (e.g., approximately 25°C), the resulting mixture is stirred or otherwise agitated for a length of time sufficient to allow salt formation to occur (e.g., 12-24 hours), and the salt is then isolated by filtration.

[0557] In a method for producing a (+)-L-tartrate salt of a tropisomer of formula (1) having pattern B (embodiment 2.13), a solution of the tropisomer in isopropyl acetate is mixed with a solution of (+)-L-tartaric acid in ethanol at a temperature in the range of 35°C to 45°C (e.g., approximately 40°C), the resulting mixture is cooled to a temperature in the range of 20°C to 30°C (e.g., approximately 25°C) over a period of approximately 1-3 hours, and the salt is then separated by filtration.

[0558] In another method for a (+)-L-tartrate salt of an atropisomer of formula (1) having pattern B (embodiment 2.14), a solution of the atropisomer in isopropyl acetate is mixed with a solution of (+)-L-tartaric acid in THF at a temperature in the range of 35°C to 45°C (e.g., approximately 40°C) (in batches or at a single feed) and one or more seed crystals of the salt of pattern B are added to produce a precipitate, the mixture is cooled to a temperature in the range of 20°C to 30°C (e.g., approximately 25°C) and stirred or agitated for a period of time (e.g., 12 to 24 hours, particularly approximately 20 hours) sufficient to allow the precipitate to mature to a state where the precipitate can be separated by filtration.

[0559] In another method for a (+)-L-tartrate salt of a tropisomer of formula (1) having pattern B (embodiment 2.15), a solution of the tropisomer in butanol is mixed (in batches or in a single feed) with a solution of (+)-L-tartaric acid in water at an elevated temperature in the range of 70°C to 85°C (e.g., approximately 80°C), the resulting mixture is cooled to an intermediate temperature in the range of 65°C to 70°C, after which one or more seed crystals of the pattern B salt are added and the mixture is cooled to a low temperature in the range of 3-10°C over a period of 8 to 15 hours, and thereafter the resulting mixture is stirred or otherwise agitated at or near low temperature for another period of 2 to 8 hours (e.g., approximately 6 hours), and the pattern B salt thus formed is then filtered out.

[0560] Protecting Group

[0561] In many of the reactions described above, it may be necessary to protect one or more groups to prevent reaction at undesired locations on the molecule. Examples of protecting groups and methods for protecting and deprotecting functional groups can be found in Protective Groups in Organic Synthesis (T. Green and P. Wuts; 3rd Edition; John Wiley and Sons, 1999).

[0562] The hydroxy group can be protected, for example, as an ether (-OR) or an ester (-OC(=O)R), for example as tert-butyl ether; tetrahydropyranyl (THP) ether; benzyl, diphenylmethyl (diphenylmethyl) or triphenylmethyl (triphenylmethyl) ether; trimethylsilyl or tert-butyldimethylsilyl ether; or as an acetyl ester (-OC(=O)CH3, -OAc).

[0563] The aldehyde or ketone group can be protected, for example, as an acetal (R-CH(OR)2) or ketal (R2C(OR)2), respectively, in which the carbonyl group (>C=O) is converted into a diether (>C(OR)2) by reaction with, for example, a primary alcohol. The aldehyde or ketone group is easily regenerated by hydrolysis using large amounts of water in the presence of an acid.

[0564] The amine group can be protected, for example, as an amide (-NRCO-R) or a carbamate (-NRCO-OR), for example as: methylamide (-NHCO-CH3); benzyloxyamide (-NHCO-OCH2C6H5, -NH-Cbz or NH-Z); as tert-butoxyamide (-NHCO-OC(CH3)3, -NH-Boc); 2-biphenyl-2-propoxyamide (-NHCO-OC(CH3)2C6H4C6H5, -NH-Bpoc), as 9-fluorenylmethoxyamide (-NH-Fmoc), as 6-nitrovaratoxyamide (-NH-Nvoc), as 2-trimethylsilylethoxyamide (-NH-Teoc), as 2,2,2-trichloroethoxyamide (-NH-Troc), as allyloxyamide (-NH-Alloc) or as 2(-phenylsulfonyl)ethoxyamide (-NH-Psec).

[0565] For example, in Scheme 1 above, when the amine H2N-YR 3 Middle part R 3When containing a second amino group, such as a cyclic amino group (e.g., piperidine or pyrrolidine group), the second amino group can be protected by means of a blocking group as described above, a preferred group being a tert-butyloxycarbonyl (Boc) group. In the case where it is not necessary to subsequently modify the second amino group, the blocking group can be loaded by this reaction sequence to obtain a compound of formula (1) in N-protected form, which can then be deprotected by standard methods (e.g., treated with acid in the case of the Boc group) to obtain a compound of formula (1).

[0566] Other protecting groups for amines, such as cyclic amines and heterocyclic NH groups, include toluenesulphonyl (tosyl) and methanesulfonyl (mesyl), benzyl groups such as p-methoxybenzyl (PMB) and tetrahydropyranyl (THP) groups.

[0567] The carboxylic acid group may be protected as an ester, for example as: C 1-7 Alkyl ester (e.g., methyl ester; tert-butyl ester); C 1-7 Haloalkyl esters (e.g., C 1-7 trihaloalkyl ester); triC 1-7 Alkylsilyl-C 1-7 Alkyl ester; or C 5-20 Aryl-C 1-7 or as an amide, for example, as a methylamide. The thiol group can be protected, for example, as a thioether (-SR), for example, as: benzyl thioether; acetamidomethyl ether (-S-CH2NHC(=O)CH3).

[0568] Isolation and purification of the compounds of the present invention

[0569] Compounds prepared by the foregoing synthetic routes can be separated and partially purified to produce mixtures of atropisomers according to standard techniques well known to those skilled in the art. One particularly useful technique in purifying compounds is preparative liquid chromatography using mass spectrometry as a means of detecting the purified compound appearing in the chromatographic column.

[0570] Preparative LC-MS is a standard and effective method for purifying small organic molecules, such as the compounds described herein. The methods of liquid chromatography (LC) and mass spectrometry (MS) can be modified to provide better separation of crude materials and improved MS detection of samples. Optimization of preparative gradient LC methods will involve changing the column, volatile eluents and modifiers, and gradients. Methods for optimizing preparative LC-MS methods and then using them to purify compounds are well known in the art. Such methods are described in Rosentreter U, Huber U.; Optimal fraction collecting in preparative LC / MS; J Comb Chem.; 2004; 6(2), 159-64 and Leister W, Strauss K, Wisnoski D, Zhao Z, Lindsley C., Development of a custom high-throughput preparative liquid chromatography / mass spectrometer platform for the preparative purification and analytical analysis of compound libraries; J Comb Chem.; 2003; 5(3); 322-9.

[0571] Examples of such systems for purifying compounds by preparative LC-MS are described below in the Examples section of this application (under the heading "Mass-Directed Purification LC-MS System"). However, it will be understood that alternative systems and methods to those described may be used. In particular, methods based on normal phase preparative LC may be used in place of the reverse phase methods described herein. Most preparative LC-MS systems use reverse phase LC and volatile acidic modifiers because the method is very effective for the purification of small molecules and because the eluent is compatible with positive ion electrospray mass spectrometry. Other chromatographic solutions, such as normal phase LC, alternatively buffered mobile phases, alkaline modifiers, etc. (as outlined in the analytical methods described below) may alternatively be used to purify compounds.

[0572] Once a mixture of atropisomers has been separated and purified to an acceptable degree, the mixture can then be subjected to a separation procedure to isolate the individual atropisomers. Thus, for example, chiral chromatography can be used to separate the individual atropisomers. The retention times of atropisomers in a chiral chromatography procedure provide a means of distinguishing between and characterizing individual atropisomers whose NMR and MS properties are generally identical.

[0573] Chiral chromatographic columns that can be used to separate individual atropisomers include immobilized chiral stationary phases (CSFs) that can be based, for example, on functionalized amylose or cellulose. Examples of such CSFs are amylose and cellulose functionalized with phenylcarbamates that have been substituted with chlorine and / or methyl. A specific example of a chiral column that can be used to separate individual atropisomers of the present invention is the "Chiralpak IG" column available from the company Daicel.

[0574] Mobile phases that can be used in conjunction with the above chiral columns typically include mixtures of: (A) liquid alkanes such as n-heptane containing a small amount (e.g., up to 1% (v / v) and more typically about 0.1% (v / v)) of an alkylamine base such as diethylamine; and (B) a mixture of alcohols and mixtures thereof such as isopropanol and methanol (e.g., 70:30 IPA:MeOH). For example, the mobile phase can include an A:B mixture in a ratio range of 80:20 to 95:5 (e.g., about 85:15 to about 90:10). The mobile phase can be used in an isocratic or gradient elution method, but in one embodiment of the invention, is used in an isocratic elution method.

[0575] The atropisomers of the present invention can also be resolved by chiral HPLC under supercritical fluid chromatography (SFC) conditions. In supercritical fluid chromatography, the mobile phase comprises a supercritical fluid such as carbon dioxide, often with a co-solvent such as an alcohol or alcohol mixture, for example methanol, ethanol and isopropanol.

[0576] The Chiralpak IG column mentioned above can be used in an SFC chromatography procedure using a carbon dioxide / methanol / isopropanol mixture as mobile phase.

[0577] Other chiral column / cosolvent combinations used in SFC include:

[0578] Lux cellulose 4 (MeOH, EtOH);

[0579] Lux cellulose 2 (MeOH);

[0580] Lux Amylose 1 (MeOH, EtOH); and

[0581] YMC Amylose-SA (MeOH, EtOH)

[0582] Lux family chiral columns are available from Phenomenex, Inc.

[0583] YMC Amylose-SA columns are available from YMC America, Inc.

[0584] Biological properties and therapeutic uses

[0585] The evidence described in the Examples below indicates that the atropisomers of the invention as defined herein are inhibitors of the polo box domains of PLK1 and PLK4 kinases, without inhibiting the catalytic domains of PLK1 and PLK4 kinases. Since the PBD domain is only present in PLK, the atropisomers should exhibit much greater selectivity (and therefore fewer unwanted side effects due to off-target kinase inhibition) than compounds that are ATP-competitive kinase inhibitors. For example, the results obtained from the studies described in Example 11F below, in which the atropisomers of formula (1) were tested against a panel of ninety-seven kinases and showed negligible activity against other kinases, demonstrate that the atropisomers of formula (1) have a high degree of selectivity for PLK1-PBD and PLK4-PBD relative to other kinases of similar structure and function. Based on this evidence, it is believed that other atropisomers of the invention, particularly those having the same R configuration as atropisomer (1), should exhibit similar advantages.

[0586] An additional advantage of inhibiting the PBD domain without inhibiting the catalytic domain is that this may result in a reduced tendency to induce drug resistance compared to PLK1 inhibitors that inhibit the catalytic domain.

[0587] The activity of the atropisomers of the invention as inhibitors of the PBD domain of PLK1 kinase can be demonstrated using the fluorescence polarization (FP) assay described in Narvaez et al., Cell Chemical Biology, 24, 1017-1028, 2017, see page 1018 and page 1026 (method details).

[0588] It is believed that the compounds of the invention may effectively exploit vulnerabilities in cellular pathways based on constitutively activated KRAS mutants, and thus the compositions of matter or atropisomers of the invention may be used to treat diseases and conditions mediated by KRAS regulation.

[0589] KRAS mutations, caused by a single nucleotide substitution, are associated with various forms of cancer. In particular, KRAS mutations are present at high rates in leukemia, colon cancer, pancreatic cancer, and lung cancer.

[0590] A preliminary screen for anticancer activity using a cancer cell line (U87MG, human brain (glioblastoma astrocytoma)) is described in Example 11A below.

[0591] In addition, it is believed that the compounds of the present invention can be used to treat cancers characterized by p53 deficiency or mutations in the TP53 gene. It is believed that PLK1 inhibits p53 in cancer cells. Therefore, after treatment with a PLK1 inhibitor, p53 in tumor cells should be activated and thus apoptosis should be induced.

[0592] It is believed that the activity of the composition of matter or atropisomer against KRAS mutant and p53-deficient cancers is at least in part produced by inhibition of PLK1 kinase, and in particular the C-terminal polo box domain (PBD) of PLK1 kinase. KRAS is known to be dependent on interaction with PLK1.

[0593] Compounds of the invention that inhibit only the PBD domain of PLK1 and not the N-terminal catalytic domain are advantageous because they are selective for PLK1-PBD over other structurally and functionally similar kinases against which these compounds show negligible inhibitory activity (see Example E below).

[0594] The compositions of matter or atropisomers of the present invention induce mitotic arrest with non-congressed chromosomes, a property believed to be caused by the PLK1-PBD and PLK4-PBD inhibitory activity of the compositions of matter or atropisomers (see Example 11C below).

[0595] The atropisomer induces mitotic arrest, has a multipolar spindle phenotype, and causes centriole enlargement, a well-described PLK4 inhibition phenotype (Lei 2018, Cell Death & Disease 9, 1066; Kawakami, PNAS 2018, 115 (8) 1913-18). These phenotypes are believed to be caused by the PLK4-PBD inhibitory activity of the atropisomer.

[0596] An additional advantage of inhibiting the PBD domain without inhibiting the catalytic domain is that this may result in a reduced tendency to induce drug resistance compared to PLK1 inhibitors that inhibit the catalytic domain.

[0597] The activity of compounds of the invention as inhibitors of the PBD domain of PLK1 kinase can be demonstrated using the fluorescence polarization (FP) assay described in Narvaez et al., Cell Chemical Biology, 24, 1017-1028, 2017, see page 1018 and page 1026 (method details).

[0598] The compounds of the invention have good oral bioavailability (see Example 11G below) and good brain exposure when orally administered (see Example 11G below). Therefore, the compositions of matter or atropisomers of the invention should be useful in treating brain cancers such as gliomas and glioblastomas.

[0599] In further embodiments (Embodiments 3.1 to 3.27), the present invention provides:

[0600] 3.1. A composition of matter, an atropisomer or a salt as defined in any one of embodiments 1.1 to 1.211 for use as a PLK1-PBD inhibitor.

[0601] 3.2 A composition of matter, an atropisomer or a salt as defined in any one of embodiments 1.1 to 1.211 for use as a PLK4-PBD inhibitor.

[0602] 3.3 A composition of matter, an atropisomer or a salt as defined in any one of embodiments 1.1 to 1.211 for use as an inhibitor of PLK1-PBD and PLK4-PBD.

[0603] 3.4 Use of a composition of matter, an atropisomer or a salt as defined in any one of Embodiments 1.1 to 1.211 for treating cancer, optionally in combination with another therapeutic agent or therapy (e.g., an anticancer agent or therapy), wherein the cancer is selected from tumors of epithelial origin (adenomas and carcinomas of various types, including adenocarcinomas, squamous cell carcinomas, transitional cell carcinomas and other carcinomas), such as bladder and urinary tract, breast, gastrointestinal tract (including esophagus, stomach (gastric), small intestine, colon, rectum and anus), liver (hepatocellular carcinoma), gallbladder and biliary system, exocrine pancreas, kidney, lung (e.g., adenocarcinoma, small cell lung cancer, non-small cell lung cancer, bronchial carcinoma and mesothelioma), head and neck (e.g., tongue, buccal cavity, larynx, pharynx, nasopharynx, tonsils, salivary glands, Cancer of the ovary, fallopian tube, peritoneum, vagina, vulva, penis, cervix, myometrium, endometrium, thyroid (e.g., follicular thyroid carcinoma), adrenal gland, prostate, skin and appendages (e.g., melanoma, basal cell carcinoma, squamous cell carcinoma, keratoacanthoma, hyperplastic nevus); hematologic malignancies (i.e., leukemias, lymphomas) and premalignant hematologic disorders and borderline malignant neoplastic disorders, including hematologic malignancies and related conditions of the lymphoid lineage (e.g., acute lymphocytic leukemia [ALL], chronic lymphocytic leukemia [CLL], B-cell lymphomas such as diffuse large B-cell lymphoma [DLBCL], follicular lymphoma, Burkitt's lymphoma, mantle cell lymphomas, T-cell lymphomas and leukemias, natural killer [NK]-cell lymphomas, Hodgkin's lymphoma, hairy cell leukemia, monoclonal gammopathy of undetermined significance, plasmacytoma, multiple myeloma, and post-transplant lymphoproliferative disorder), as well as hematologic malignancies of the myeloid lineage and related conditions (e.g., acute myeloid leukemia [AML], chronic myeloid leukemia [CML], chronic myelomonocytic leukemia [CMML], hypereosinophilic syndromes, myeloproliferative disorders such as polycythemia vera, essential thrombocythaemia and primary myelofibrosis, myeloproliferative syndromes, myelodysplastic syndromes, and promyelocytic leukemias) Tumors of mesenchymal origin, for example sarcomas of soft tissue, bone or cartilage, such as osteosarcoma, fibrosarcoma, chondrosarcoma, rhabdomyosarcoma, leiomyosarcoma, liposarcoma, angiosarcoma, Kaposi's sarcoma, Ewing's sarcoma, synovial sarcoma, epithelioid sarcoma, gastrointestinal stromal tumors, benign and malignant histiocytomas and dermatofibrosarcoma protuberans; central or peripheral nervous system tumors (e.g. astrocytomas, gliomas and glioblastomas, meningiomas, ependymomas, pinealomas and schwannomas); endocrine tumors (e.g. pituitary tumors, adrenal tumors, islet cell tumors, parathyroid tumors, carcinoid and medullary carcinoma of the thyroid gland); eye and adnexal tumors (e.g. retinoblastoma);Germ cell and trophoblastic tumors (e.g., teratoma, seminoma, dysgerminoma, hydatidiform mole, and choriocarcinoma); and pediatric and embryonal tumors (e.g., medulloblastoma, neuroblastoma, Wilms tumor, and primitive neuroectodermal tumor); or congenital or other syndromes that predispose patients to malignancy (e.g., xeroderma pigmentosum).

[0604] 3.5 Use of a composition of matter, an atropisomer or a salt as defined in any one of embodiments 1.1 to 1.211 for treating cancer, optionally in combination with another therapeutic agent or therapy (e.g. an anticancer agent or therapy), wherein the cancer is selected from pancreatic cancer, colon cancer and colorectal cancer, lung cancer, brain cancer and neurological cancer and blood cancers such as lymphoma and leukemia.

[0605] 3.6 Use of a composition of matter, an atropisomer or a salt as defined in any one of embodiments 1.1 to 1.211 for treating cancer, optionally in combination with another therapeutic agent or therapy (e.g., an anticancer agent or therapy), wherein the cancer is selected from glioma and glioblastoma (which may, for example, be selected from glioblastoma multiforme, ependymoma, diffuse intrinsic pontine glioma, IDH1 mutant glioma).

[0606] 3.7 Use of a composition of matter, an atropisomer or a salt as defined in any one of embodiments 1.1 to 1.211 for treating cancer, optionally in combination with another therapeutic agent or therapy (e.g., an anticancer agent or therapy), wherein the cancer is selected from rhabdoid tumor; medulloblastoma and other embryonal tumors of the brain; breast cancer, lung cancer, melanoma, gastric cancer, colorectal cancer, pancreatic cancer and ovarian cancer.

[0607] 3.8 Use of a composition of matter, an atropisomer or a salt as defined in any one of Embodiments 1.1 to 1.211 for treating cancer, optionally in combination with another therapeutic agent or therapy (e.g., an anti-cancer agent or therapy), wherein the cancer is a cancer in which PLK1 is implicated (e.g., in which PLK1 is overexpressed).

[0608] 3.9 A composition of matter, atropisomer or salt for use according to Embodiment 3.8, wherein the cancer is as defined in any one of Embodiments 3.4 to 3.7.

[0609] 3.10 A composition of matter, an atropisomer or a salt as defined in any one of Embodiments 1.1 to 1.211 for use in treating cancer, optionally in combination with another therapeutic agent or therapy (e.g., an anti-cancer agent or therapy), wherein the cancer is a cancer in which PLK4 is implicated (e.g., in which PLK4 is overexpressed).

[0610] 3.11 A composition of matter, atropisomer or salt for use according to Embodiment 3.10, wherein the cancer is as defined in any one of Embodiments 3.4 to 3.7.

[0611] 3.12 Use of a composition of matter, an atropisomer or a salt as defined in any one of embodiments 1.1 to 1.211 for treating cancer, optionally in combination with another therapeutic agent or therapy (e.g., an anti-cancer agent or therapy), wherein the cancer is a cancer characterized by a p53 defect or a mutation in the TP53 gene.

[0612] 3.13 A composition of matter, atropisomer or salt for use according to Embodiment 3.12 wherein the cancer is as defined in any one of Embodiments 3.4 to 3.7.

[0613] 3.14 A composition of matter, an atropisomer or a salt as defined in any one of Embodiments 1.1 to 1.211 for use in treating cancer, wherein the cancer is characterised by the presence of a mutant form of KRAS.

[0614] 3.15 A composition of matter, atropisomer or salt for use according to Embodiment 3.14, wherein the mutant form of KRAS is a mutant form having a mutation at an amino acid in the protein selected from glycine 12, glycine 13, glutamine 61 and combinations thereof.

[0615] 3.16 A composition of matter, atropisomer or salt for use according to Embodiment 3.14 or 3.15 wherein the cancer is as defined in any one of Embodiments 3.4 to 3.7.

[0616] 3.17 A composition of matter, an atropisomer or a salt as defined in any one of Embodiments 1.1 to 1.211 for use in medicine or therapy, optionally in combination with another therapeutic agent or therapy (eg an anti-cancer agent or therapy).

[0617] 3.18 A composition of matter, atropisomer or salt as defined in any one of Embodiments 1.1 to 1.211 for use in preventing or treating disease states and disorders characterised by aberrant expression of the KRAS protein, optionally in combination with another therapeutic agent or therapy (eg, an anti-cancer agent or therapy).

[0618] 3.19 A composition of matter, an atropisomer or a salt as defined in any one of Embodiments 1.1 to 1.211 for use as an anticancer agent.

[0619] 3.20 A method of treating a subject (e.g., a mammalian subject such as a human) having a cancer as defined in any one of Embodiments 3.4 to 3.16, the method comprising administering to the subject a therapeutically effective amount of a composition of matter, atropisomer or salt as defined in any one of Embodiments 1.1 to 1.211, optionally in combination with another therapeutic agent or therapy (e.g., an anti-cancer agent or therapy).

[0620] 3.21 Use of a composition of matter, atropisomer or salt as defined in any one of Embodiments 1.1 to 1.211 for the manufacture of a medicament for a use as defined in any one of Embodiments 3.1 to 3.19.

[0621] 3.22 A method of inhibiting PLK1-PBD, the method comprising contacting an effective kinase inhibiting amount of a composition of matter, atropisomer or salt as defined in any one of Embodiments 1.1 to 1.211 with the PLK1-PBD.

[0622] 3.23 A method of inhibiting PLK1 kinase, the method comprising contacting the PLK1 kinase with a kinase inhibiting amount of a composition of matter, atropisomer or salt as defined in any one of Embodiments 1.1 to 1.211.

[0623] 3.24 A method of inhibiting PLK4-PBD, the method comprising contacting an effective inhibitory amount of a composition of matter, atropisomer or salt as defined in any one of Embodiments 1.1 to 1.211 with the PLK4-PBD.

[0624] 3.25 A method of inhibiting PLK4 kinase, the method comprising contacting the PLK4 kinase with a kinase inhibiting amount of a composition of matter, atropisomer or salt as defined in any one of Embodiments 1.1 to 1.211.

[0625] 3.26 A method of inhibiting PLK1-PBD and PLK4-PBD, the method comprising contacting the PLK1-PBD and PLK4-PBD with an effective inhibitory amount of a composition of matter, atropisomer or salt as defined in any one of Embodiments 1.1 to 1.211.

[0626] 3.27 A method according to any one of embodiments 3.22 to 3.26, wherein the effective inhibitory amount of a composition of matter, an atropisomer or a salt as defined in any one of embodiments 1.1 to 1.211 is contacted with the PLK1-PBD and / or PLK4-PBD in vivo, for example in a mammalian subject such as a human subject.

[0627] Prior to administering a composition of matter, atropisomer or salt as defined in any one of Embodiments 1.1 to 1.211, a patient may be screened to determine whether the cancer the patient has or may have is a cancer characterized by elevated levels of PLK1 and / or PLK4 kinase and therefore would be sensitive to treatment with a compound having activity against PLK1 and / or PLK4 kinase.

[0628] For example, a biological sample taken from a patient can be analyzed to determine whether the cancer that the patient suffers from or may suffer from is a cancer characterized by genetic abnormalities or abnormal protein expression (leading to upregulation of PLK1 and / or PLK4 kinases). The term upregulation includes elevated expression or overexpression, including gene amplification (i.e., multiple gene copies) and increased expression caused by transcription, as well as overactivity and activation, including mutation activation. Therefore, a patient can be subjected to a diagnostic test to detect markers characteristic of upregulation of PLK1 and / or PLK4 kinases. The term diagnosis includes screening. For markers, we include genetic markers (e.g., including measuring DNA composition) to identify mutations of PLK1 and / or PLK4 kinases. The term marker also includes markers characterized by upregulation of PLK1 and / or PLK4, including the enzyme activity, enzyme level, enzyme state (e.g., phosphorylation or not) and mRNA level of the aforementioned proteins.

[0629] Tumors with upregulated PLK1 and / or PLK4 kinases may be particularly sensitive to PLK1 inhibitors. Screening for tumors with upregulated PLK1 and / or PLK4 may be preferred. Thus, patients may be subjected to diagnostic tests to detect markers characteristic of upregulated PLK1 and / or PLK4. Diagnostic tests are typically performed on biological samples selected from tumor biopsy samples, blood samples (isolation and enrichment of exfoliated tumor cells), stool biopsies, sputum, chromosome analysis, pleural fluid, and peritoneal fluid.

[0630] Methods for identifying and analyzing protein mutations and upregulation are known to those skilled in the art. Screening methods may include, but are not limited to, standard methods such as reverse transcriptase polymerase chain reaction (RT-PCR) or in situ hybridization.

[0631] In screening by RT-PCR, the level of mRNA in the tumor is assessed by generating a cDNA copy of the mRNA and then amplifying the cDNA by PCR. The method of PCR amplification, the selection of primers and the conditions for amplification are known to those skilled in the art. Nucleic acid manipulation and PCR are performed by standard methods, such as described, for example, in Ausubel, FM et al., eds. Current Protocols in Molecular Biology, 2004, John Wiley & Sons Inc., or Innis, MA et-al., eds. PCR Protocols: a guide to methods and applications, 1990, Academic Press, San Diego. Reactions and manipulations involving nucleic acid techniques are also described in Sambrook et al., 2001, 3 rd Ed, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press. Alternatively, commercially available kits for RT-PCR (e.g., Roche Molecular Biochemicals) or methods as listed in U.S. Pat. Nos. 4,666,828; 4,683,202; 4,801,531; 5,192,659, 5,272,057, 5,882,864, and 6,218,529 and incorporated herein by reference may be used.

[0632] An example of an in situ hybridization technique for assessing mRNA expression would be fluorescence in situ hybridization (FISH) (see Angerer, 1987 Meth. Enzymol., 152:649).

[0633] Typically, in situ hybridization includes the following major steps: (1) fixation of the tissue to be analyzed; (2) pre-hybridization treatment of the sample to increase accessibility of the target nucleic acid and reduce nonspecific binding; (3) hybridization of the nucleic acid mixture with nucleic acids in the biological structure or tissue; (4) post-hybridization washing to remove nucleic acid fragments that are not bound in the hybridization, and (5) detection of the hybridized nucleic acid fragments. Probes used in such applications are typically labeled with, for example, radioactive isotopes or fluorescent reporter molecules. Preferred probes are sufficiently long, for example, from about 50, 100 or 200 nucleotides to about 1000 or more nucleotides, to enable specific hybridization to one or more target nucleic acids under stringent conditions. Standard methods for performing FISH are described in Ausubel, F Metal., eds. Current Protocols in Molecular Biology, 2004, John Wiley & Sons Inc and Fluorescence In Situ Hybridization: Technical Overview by John MS Bartlett in Molecular Diagnosis of Cancer, Methods and Protocols, 2nd ed.; ISBN: 1-59259-760-2; March 2004, pps.077-088; Series: Methods in Molecular Medicine.

[0634] Alternatively, the protein product expressed from the mRNA can be determined by immunohistochemistry of tumor samples, solid phase immunoassays in microtiter plates, Western blotting, 2-dimensional SDS-polyacrylamide gel electrophoresis, ELISA, flow cytometry, and other methods known in the art for detecting specific proteins. Detection methods will include the use of site-specific antibodies. The skilled artisan will recognize that all such well-known techniques for detecting upregulation of PLK1 and / or PLK4 kinases can be adapted in the present case.

[0635] Alternatively or additionally, prior to administering a composition of matter, atropisomer or salt as defined in any one of Embodiments 1.1 to 1.211, a patient may be screened to determine whether the cancer the patient has or may have is a cancer characterised by mutant KRAS and therefore would be sensitive to treatment with a compound having activity against cancer cells carrying mutant KRAS.

[0636] For example, a biological sample taken from a patient can be analyzed to determine whether the cancer that the patient has or may have is a cancer characterized by the presence of mutant KRAS. Thus, for example, a diagnostic test can be performed on a patient to detect mutations at codons 12, 13, 61 (glycine 12, glycine 13, and glutamine 61) in the KRAS protein, or a mixture thereof. Commercially available diagnostic tests for mutant KRAS include the ELISA kit from Roche Molecular Systems, Inc. KRAS mutation testing and therascreen KRAS RGQ PCR kit from Qiagen Manchester, Ltd.

[0637] Tumors with mutant KRAS may be particularly sensitive to PLK1 and / or PLK4 inhibitors. Methods for identifying and analyzing protein mutations and upregulation are known to those skilled in the art. Screening methods may include, but are not limited to, standard methods such as reverse transcriptase polymerase chain reaction (RT-PCR) or in situ hybridization as described above.

[0638] Therefore, in further embodiments (Embodiments 3.28 to 3.38), the present invention provides:

[0639] 3.28 Use of a composition of matter, an atropisomer or a salt as defined in any one of Embodiments 1.1 to 1.211 for treating cancer in a subject (e.g., a human subject) who has been screened and determined to have a cancer characterised by elevated PLK1 kinase levels (e.g., PLK1 overexpression).

[0640] 3.29 Use of a composition of matter, an atropisomer or a salt as defined in any one of Embodiments 1.1 to 1.211 for treating cancer in a subject (e.g., a human subject) who has been screened and determined to have a cancer characterised by elevated PLK4 kinase levels (e.g., PLK4 overexpression).

[0641] 3.30 Use of a composition of matter, an atropisomer or a salt as defined in any one of Embodiments 1.1 to 1.211 for treating cancer in a subject (e.g., a human subject) who has been screened and determined to have a cancer characterized by elevated levels of PLK1 kinase and PLK4 kinase (e.g., overexpression of PLK1 and PLK4).

[0642] 3.31 Use of a composition of matter, an atropisomer or a salt as defined in any one of Embodiments 1.1 to 1.211 for treating cancer in a subject (e.g., a human subject) who has been screened and determined to have, or be at risk of having, a disease or condition that would be sensitive to treatment with a compound having activity against KRAS.

[0643] 3.32 Use of a composition of matter, an atropisomer or a salt as defined in any one of Embodiments 1.1 to 1.211 for treating a subject (e.g., a human subject) who has been screened and determined to have a cancer that is characterized by mutant KRAS and would be sensitive to treatment with a compound that has activity against KRAS or against cancer cells carrying mutant KRAS.

[0644] 3.33 A composition of matter, atropisomer or salt for use according to any one of Embodiments 3.28 to 3.32 wherein the cancer is a cancer as defined in any one of Embodiments 3.4 to 3.16.

[0645] 3.34 Use of a composition of matter, atropisomer or salt as defined in any one of Embodiments 1.1 to 1.211 for the manufacture of a medicament for a use as defined in any one of Embodiments 3.28 to 3.33.

[0646] 3.35 A method for diagnosing and treating a disease state or condition mediated by KRAS or characterized by the presence of a mutant form of KRAS (e.g., cancer, e.g., a cancer as defined in any one of Embodiments 3.4 to 3.16), the method comprising (i) screening a subject (e.g., a human subject) to determine whether the disease or condition that the subject suffers from or may suffer from is a disease or condition that would be sensitive to treatment with a compound having activity against KRAS; and (ii) where the disease or condition that the subject suffers from is indicated to be therefore sensitive, thereafter administering to the subject a therapeutically effective amount of a composition of matter, atropisomer or salt as defined in any one of Embodiments 1.1 to 1.211.

[0647] 3.36 A method for treating a disease state or condition characterized by the presence of a mutant form of KRAS (e.g., cancer, e.g., a cancer as defined in any one of Embodiments 3.4 to 3.16), the method comprising administering to a subject (e.g., a human subject) a therapeutically effective amount of a composition of matter, atropisomer or salt as defined in any one of Embodiments 1.1 to 1.211, the subject having been screened and determined to have, or be at risk of having, a disease or condition that would be susceptible to treatment with a compound having activity against KRAS.

[0648] 3.37 A method for diagnosing and treating cancer characterized by elevated PLK1 kinase levels, the method comprising (i) screening a patient to determine whether the cancer suffered by the patient is a cancer characterized by elevated PLK1 kinase levels; and (ii) where the cancer is indicated to be a cancer characterized by elevated PLK1 kinase levels, thereafter administering to the patient an effective amount of a composition of matter, a transposable isomer or a salt as defined in any one of Embodiments 1.1 to 1.211.

[0649] 3.38 Use of a composition of matter, atropisomer or salt as defined in any one of Embodiments 1.1 to 1.211 for the manufacture of a medicament for treating or preventing a disease state or condition in a patient who has been screened and determined to have, or to be at risk of having, a disease or condition that would be sensitive to treatment with a compound having activity against KRAS.

[0650] Pharmaceutical preparations

[0651] The composition of matter or atropisomer of the present invention is usually administered to a patient in the form of a pharmaceutical composition. Therefore, in another embodiment of the present invention (embodiment 4.1), the present invention provides a pharmaceutical composition comprising a composition of matter as defined in any one of embodiments 1.1 to 1.211, an atropisomer or a salt and a pharmaceutically acceptable excipient.

[0652] In another embodiment, there is provided:

[0653] 4.2 A pharmaceutical composition according to embodiment 4.1, comprising from about 1% (w / w) to about 95% (w / w) of a composition of matter as defined in any one of embodiments 1.1 to 1.211, an atropisomer or a salt and from 99% (w / w) to 5% (w / w) of a pharmaceutically acceptable excipient or combination of excipients and optionally one or more additional therapeutically active ingredients.

[0654] 4.3 A pharmaceutical composition according to embodiment 4.2, comprising from about 5% (w / w) to about 90% (w / w) of a composition of matter as defined in any one of embodiments 1.1 to 1.211, an atropisomer or a salt and from 95% (w / w) to 10% of a pharmaceutical excipient or a combination of excipients and optionally one or more additional therapeutically active ingredients.

[0655] 4.4 A pharmaceutical composition according to embodiment 4.3, comprising from about 10% (w / w) to about 90% (w / w) of a composition of matter, an atropisomer or a salt as defined in any one of embodiments 1.1 to 1.211 and from 90% (w / w) to 10% of a pharmaceutical excipient or a combination of excipients.

[0656] 4.5 A pharmaceutical composition according to embodiment 4.4, comprising about 20% (w / w) to about 90% (w / w) of a composition of matter, an atropisomer or a salt as defined in any one of embodiments 1.1 to 1.211 and 80% (w / w) to 10% of a pharmaceutical excipient or a combination of excipients.

[0657] 4.6 A pharmaceutical composition according to embodiment 4.5, comprising about 25% (w / w) to about 80% (w / w) of a composition of matter, an atropisomer or a salt as defined in any one of embodiments 1.1 to 1.211 and 75% (w / w) to 20% of a pharmaceutical excipient or a combination of excipients.

[0658] The pharmaceutical compositions of the present invention may be in any form suitable for oral, parenteral, topical, intranasal, intrabronchial, intraocular, auricular, rectal, intravaginal or transdermal administration. When the compositions are intended for parenteral administration, they may be formulated for intravenous, intramuscular, intraperitoneal, subcutaneous administration or delivered directly to a target organ or tissue by injection, infusion or other delivery methods.

[0659] Suitable pharmaceutical dosage forms for oral administration include tablets, capsules, caplets, pills, lozenges, syrups, solutions, sprays, powders, granules, elixirs and suspensions, sublingual tablets, sprays, wafers or patches and buccal patches.

[0660] Therefore, in further embodiments, the present invention provides:

[0661] 4.7 A pharmaceutical composition according to any one of Embodiments 4.1 to 4.6, which is suitable for oral administration.

[0662] 4.8 A pharmaceutical composition according to embodiment 4.7 which is selected from tablets, capsules, caplets, pills, lozenges, syrups, solutions, sprays, powders, granules, elixirs and suspensions, sublingual tablets, sprays, wafers or patches and buccal patches.

[0663] 4.9 A pharmaceutical composition according to Embodiment 4.8 which is selected from the group consisting of a tablet and a capsule.

[0664] 4.10 A pharmaceutical composition according to any one of Embodiments 4.1 to 4.6 which is suitable for parenteral administration.

[0665] 4.11 A pharmaceutical composition according to Embodiment 4.10, which is formulated for intravenous, intramuscular, intraperitoneal, subcutaneous administration or direct delivery into a target organ or tissue by injection, infusion or other means of delivery.

[0666] 4.12 A pharmaceutical composition according to Embodiment 4.11 which is a solution or suspension for injection or infusion.

[0667] A pharmaceutical composition comprising a composition of matter, atropisomer or salt as defined in any one of Embodiments 1.1 to 1.211 of the invention (e.g., as defined in any one of Embodiments 4.1 to 4.12) can be formulated according to known techniques, see, e.g., Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA, USA.

[0668] Thus, tablet compositions (such as in embodiment 4.9) may contain a unit dose of active compound together with an inert diluent or carrier, such as a sugar or sugar alcohol, for example; lactose, sucrose, sorbitol or mannitol; and / or a non-sugar derived diluent, such as sodium carbonate, calcium phosphate, talc, calcium carbonate, or cellulose or its derivatives, such as methylcellulose, ethylcellulose, hydroxypropylmethylcellulose, and starch, such as corn starch. Tablets may also contain standard ingredients, such as binders and granulating agents (such as polyvinylpyrrolidone), disintegrants (e.g. swellable cross-linked polymers, such as cross-linked carboxymethylcellulose), lubricants (e.g. stearates), preservatives (e.g. parabens), antioxidants (e.g. BHT), buffers (e.g. phosphate or citrate buffers) and effervescent agents (such as citrate / bicarbonate mixtures). Such excipients are well known and need not be discussed in detail here.

[0669] Capsule formulations (such as in embodiment 4.9) may be of hard or soft gelatin variety and may contain the active ingredient in solid, semisolid or liquid form.Gelatin capsules may be formed from animal gelatin or its equivalent of synthetic or vegetable origin.

[0670] Solid dosage forms (e.g., tablets, capsules, etc.) may be coated or uncoated, but typically have a coating, such as a protective film coating (e.g., wax or varnish) or a controlled release coating. Coatings (e.g., Eudragit TM The coating can be designed to release the active ingredient at a desired location in the gastrointestinal tract. Thus, the coating can be selected to degrade under certain pH conditions in the gastrointestinal tract, thereby selectively releasing the composition of matter or atropisomer in the stomach or ileum or duodenum.

[0671] Instead of or in addition to a coating, the drug may be presented in a solid matrix including a controlled release agent (e.g., a release delaying agent) which may be adapted to selectively release the composition of matter or atropisomer under conditions of varying acidity or alkalinity in the gastrointestinal tract. Alternatively, the matrix material or delayed release coating may take the form of an erodible polymer (e.g., a maleic anhydride polymer) which is substantially continuously eroded as the dosage form passes through the gastrointestinal tract.

[0672] Compositions for external use include ointments, creams, sprays, patches, gels, liquid drops and inserts (eg, intraocular inserts). Such compositions can be formulated according to known methods.

[0673] Compositions for parenteral administration (such as those in Embodiments 4.10 to 4.12) are generally presented as sterile aqueous or oily solutions or fine suspensions, or may be provided in the form of finely divided sterile powders for extemporaneous preparation with sterile water for injection.

[0674] Examples of formulations for rectal or vaginal administration include pessaries and suppositories which may, for example, be formed from shaped moldable or waxy materials containing the active compound.

[0675] Compositions for administration by inhalation can take the form of inhalable powder compositions or liquid or powder sprays and can be administered in standard form using a powder inhaler device or an aerosol dispensing device. Such devices are well known. For administration by inhalation, powdered preparations typically include the active compound together with an inert solid powdered diluent, such as lactose.

[0676] The compositions of matter or atropisomers of the invention will typically be presented in unit dosage form, and as such will typically contain sufficient compound to provide the desired level of biological activity. For example, according to any one of Embodiments 3.1 to 3.9, a composition intended for oral administration may contain 2 mg to 200 mg of active ingredient, more typically 10 mg to 100 mg, such as 12.5 mg, 25 mg, and 50 mg.

[0677] Dosimetry

[0678] The active compound (a composition of matter, atropisomer or salt as defined in any one of Embodiments 1.1 to 1.211) will be administered to a patient (e.g., a human or animal patient) in need thereof in an amount sufficient to achieve the desired therapeutic effect (e.g., an effect as described above in Embodiments 3.1 to 3.38).

[0679] The composition of matter, atropisomer or salt is typically administered to a subject in need of such administration, such as a human or animal patient, preferably a human.

[0680] The composition of matter, atropisomer or salt is generally administered in a therapeutically or prophylactically useful and generally non-toxic amount. However, in certain circumstances, the benefits of administering the composition of matter, atropisomer or salt may outweigh the disadvantages of any toxic effects or side effects, in which case it may be considered desirable to administer the compound in an amount associated with the degree of toxicity.

[0681] A typical daily dosage of the composition of matter, atropisomer or salt may be in the range of 0.025 mg to 5 mg per kg body weight, for example up to 3 mg per kg body weight, and more usually 0.15 mg to 5 mg per kg body weight, although higher or lower doses may be administered if desired.

[0682] By way of example, an initial starting dose of 12.5 mg may be administered 2 to 3 times a day. The dose may be increased by 12.5 mg a day every 3 to 5 days until the individual maximum tolerated and effective dose as determined by the physician is reached. Ultimately, the amount of compound administered should be commensurate with the nature of the disease or physiological condition being treated and the therapeutic benefit and presence or absence of side effects resulting from a given dosage regimen, and should be determined by the physician.

[0683] Combination therapy

[0684] It is envisaged that a composition of matter, atropisomer or salt as defined in any one of Embodiments 1.1 to 1.211 may be used as a sole chemotherapeutic agent, or more generally, in combination therapy with a chemotherapeutic agent or radiotherapy for the prevention or treatment of a range of proliferative disease states or disorders. Examples of such disease states and disorders are listed above.

[0685] Specific examples of chemotherapeutic agents or other therapies that can be co-administered with a composition of matter, atropisomer or salt as defined in any one of Embodiments 1.1 to 1.211:

[0686] Topoisomerase I inhibitors

[0687] Antimetabolites: (e.g. cytarabine)

[0688] Tubulin targeting agents

[0689] DNA binding agents and topoisomerase II inhibitors

[0690] EGFR inhibitors (eg, gefitinib – see Biochemical Pharmacology 78 2009 460-468)

[0691] mTOR inhibitors (eg, Everolimus)

[0692] PI3K pathway inhibitors (e.g. PI3K, PDK1)

[0693] Akt inhibitors

[0694] Alkylating agents (eg, temozolomide)

[0695] Monoclonal antibodies.

[0696] Anti-hormones

[0697] Signal transduction inhibitors

[0698] Proteasome inhibitors

[0699] DNA methyltransferase inhibitors

[0700] Cytokines and retinoids

[0701] Hypoxia-induced DNA damage agents (e.g. Tirapazamine)

[0702] Aromatase inhibitors

[0703] Anti-Her2 antibodies (see, e.g., http: / / www.wipo.int / pctdb / en / wo.jsp?wo=2007056118)

[0704] Anti-CD20 antibody

[0705] Angiogenesis inhibitors

[0706] HDAC inhibitors

[0707] MEK inhibitors

[0708] B-Raf inhibitors

[0709] ERK inhibitors

[0710] Small molecule HER2 inhibitors, such as lapatinib

[0711] Bcr-Abl tyrosine kinase inhibitors such as imatinib

[0712] CDK4 / 6 inhibitors, such as palbociclib (Ibrance)

[0713] Mps1 / TTK inhibitors

[0714] Aurora B inhibitors

[0715] FLT3 kinase inhibitors

[0716] IDH1 or IDH2 inhibitors

[0717] BRD4 inhibitors

[0718] Temozolomide

[0719] Inhibitors of immune checkpoint blockade signaling components, including PD1, PDL-1, and CTLA4; and

[0720] Radiation therapy.

[0721] Therefore, in further embodiments, the present invention provides:

[0722] 5.1 A pharmaceutical combination comprising a composition of matter as defined in any one of Embodiments 1.1 to 1.211, an atropisomer or a salt and another therapeutically active agent.

[0723] 5.2 The pharmaceutical combination according to Embodiment 5.1, wherein the further therapeutic agent is selected from the chemotherapeutic agents listed above.

[0724] 5.3 The pharmaceutical combination according to Embodiment 5.1, wherein the other therapeutic agent is an anti-cancer agent.

[0725] 5.4 A pharmaceutical combination according to any one of embodiments 5.1 to 5.3, wherein the composition of matter, atropisomer or salt as defined in any one of embodiments 1.1 to 1.211 and the further therapeutically active agent are presented in a single pharmaceutical composition or patient pack.

[0726] 5.5 A pharmaceutical composition comprising a composition of matter, an atropisomer or a salt as defined in any one of Embodiments 1.1 to 1.211, another therapeutically active agent and at least one pharmaceutically acceptable excipient.

[0727] 5.6 A method of treating a subject having cancer, the method comprising administering to the subject a therapeutically effective amount of the pharmaceutical combination according to any one of Embodiments 5.1 to 5.5.

[0728] 5.7 Use of a composition of matter, an atropisomer or a salt as defined in any one of Embodiments 1.1 to 1.211 for enhancing the therapeutic effect of radiotherapy or chemotherapy in the treatment of a proliferative disease such as cancer.

[0729] 5.8 Use of a composition of matter, an atropisomer or a salt as defined in any one of Embodiments 1.1 to 1.211, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for enhancing the therapeutic effect of radiotherapy or chemotherapy in the treatment of a proliferative disease such as cancer.

[0730] 5.9 A method for preventing or treating a proliferative disease such as cancer, the method comprising administering to a patient a composition of matter, an atropisomer or a salt as defined in any one of Embodiments 1.1 to 1.211, or a pharmaceutically acceptable salt thereof, in combination with radiation therapy or chemotherapy. BRIEF DESCRIPTION OF THE DRAWINGS

[0731] Figure 1 is a schematic diagram illustrating the R / S classification system for atropisomers.

[0732] Figure 2 is a depiction of the three-dimensional structure of 4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]benzamide atropisomer A-2 as determined by single crystal X-ray crystallographic studies.

[0733] Figure 3 It is the basis for the schematic stereochemical representation of the two atropisomers A-1 (S) and A-2 (R) and the assignment of their stereochemical structures using the Cahn–Ingold–Prelog (CIP) sequential rules.

[0734] Figure 4 is an X-ray powder diffraction spectrum of atropisomer A-2 free base.

[0735] FIG5 is an X-ray powder diffraction spectrum of atropisomer A-2 tartrate pattern A salt (bottom line) and pattern B salt (top line and middle line)

[0736] Figure 6 illustrates the thermal profile of atropisomer A-2 free base and shows a differential scanning calorimetry plot (line 6A) and a thermogravimetric analysis plot (line 6B).

[0737] Figure 7 illustrates the thermal profile of atropisomer A-2 tartrate pattern A salt and shows a differential scanning calorimetry plot (line 7A) and a thermogravimetric analysis plot (line 7B).

[0738] Figure 8 illustrates the thermal profile of atropisomer A-2 tartrate pattern B salt and shows a differential scanning calorimetry plot (line 8A) and a thermogravimetric analysis plot (line 8B).

[0739] Fig. 9 is a plot of weight change versus relative humidity from a gravimetric vapor sorption study conducted on atropisomer A-2 tartrate pattern B salt.

[0740] Fig.10is a bar graph showing the proportions of different mitotic phenotypes (non-dividing chromosomes, multipolar spindle / abnormal cytokinesis, monopolar spindle, normal prometaphase, and subsequent normal metaphase) observed after treatment of U87MG cells with either atropisomer A-1 or atropisomer A-2 at a concentration of 0.03 μM.

[0741] Fig.11 is a bar graph showing the number of centrioles present in HeLa cells after treatment with either atropisomer A-1 or atropisomer A-2 at a concentration of 0.02 μM.

[0742] Fig.12 is a graph of plasma concentration versus time following oral and iv administration of atropisomer A-2 to mice. The lower line extending to 24 hours is the line for 2 mg / kg iv administration. The other line is the line for 10 mg / kg po administration.

[0743] Fig.13 is a graph of plasma concentration versus time following oral and iv administration of atropisomer A-3 to mice. The lower line extending to 24 hours is the line for iv administration. The other line is the line for po administration.

[0744] Fig.14 is a graph of plasma and brain concentrations versus time following oral administration (10 mg / kg) of atropisomer A-2 to mice. The upper line shows brain concentrations and the lower line shows plasma concentrations.

[0745] Fig.15 is a graph of plasma and brain concentrations versus time following oral administration of atropisomer A-3 to mice. The upper line shows brain concentrations and the lower line shows plasma concentrations.

[0746] Fig.16 is a graph of tumor volume versus time in male athymic nude mice in the U87MG subcutaneous xenograft model following administration of atropisomer A-2.

[0747] Fig.17 is a graphical comparison of bioluminescent signals associated with tumor growth in male athymic nude mice in the U87-Luc orthotopic xenograft model following administration of atropisomer A-2.

[0748] Fig.18 is a graph of tumor volume versus time in male athymic nude mice in the HCT116 subcutaneous xenograft model following administration of atropisomer A-2.

[0749] Fig.19 XRPD patterns of atropisomer A-2 hydrochloride salt patterns A and B are shown.

[0750] Fig. 20Shown is the XRPD pattern of atropisomer A-2 mesylate salt.

[0751] Fig.21 XRPD patterns of atropisomer A-2 maleate salt patterns A and B are shown.

[0752] Fig. 22 XRPD patterns of atropisomer A-2 malate, patterns A and B, are shown.

[0753] Fig.23 XRPD pattern of atropisomer A-2 tosylate salt, pattern A, is shown.

[0754] Fig.24 XRPD patterns of atropisomer A-2 phosphate patterns A and B are shown.

[0755] Fig.25 XRPD patterns of atropisomer A-2 sulfate salt patterns A and B are shown. Example

[0756] The present invention will now be illustrated, but not limited, by reference to the specific embodiments described in the following examples.

[0757] In the examples, the following abbreviations are used.

[0758] aq aqueous / aqueous solution

[0759] CaCl2 Calcium Chloride

[0760] DCM Dichloromethane

[0761] DEA Diethylamine

[0762] DIPEA N,N-Diisopropylethylamine

[0763] DMF Dimethylformamide

[0764] DMP Dess-Martin Periodinane

[0765] DMSO Dimethyl sulfoxide

[0766] Et2O Diethyl ether

[0767] EtOAc Ethyl acetate

[0768] EtOH

[0769] h hour

[0770] HATU (1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide hexafluorophosphate)

[0771] HCl Hydrogen chloride

[0772] HPLC High Performance Liquid Chromatography

[0773] H2SO4 sulfuric acid

[0774] IPA Isopropyl Alcohol

[0775] LC Liquid Chromatography

[0776] LCMS liquid chromatography-mass spectrometry

[0777] LiOH Lithium Hydroxide

[0778] MeCN Acetonitrile

[0779] MeOH Methanol

[0780] min

[0781] MTBE Methyl tert-butyl ether

[0782] NaBH4 Sodium borohydride

[0783] NaHCO3 Sodium bicarbonate

[0784] NaOH Sodium hydroxide

[0785] Na2SO4 Sodium sulfate

[0786] NH4Cl Ammonium chloride

[0787] NMR Nuclear Magnetic Resonance

[0788] PTSA p-Toluenesulfonic acid

[0789] TEA Triethylamine

[0790] THF Tetrahydrofuran

[0791] Compound details and experiments

[0792] Atropisomers A-1 to A-8

[0793]

[0794]

[0795] Unless otherwise stated, proton magnetic resonance (1H NMR) spectra were recorded on a Bruker 400 instrument operating at 400 MHz in DMSO-d6 or MeOH-d4 (as indicated) at 27°C and reported as follows: chemical shift δ / ppm (multiplicity, where s=singlet, d=doublet, dd=doublet of doublets, dt-doublet of triplets, t=triplet, q=quartet, m=multiplet, br=broad, number of protons). Residual protic solvent was used as an internal reference.

[0796] Liquid chromatography and mass spectrometry analyses were performed using the systems and operating conditions listed below. If atoms with different isotopes are present and a single mass is quoted, the mass quoted for that compound is the monoisotopic mass (i.e. 35 Cl; 79 Br, etc.).

[0797] LCMS conditions

[0798] The LCMS data presented in the following examples were obtained using one of the methods described below.

[0799] HPLC Method 1

[0800] LCMS was performed on a UPLC AQUITY with a PDA photodiode array detector and a QDa mass detector. The column used was C18, 2.1x50mm, 1.9μm. The column flow was 1.2mL / min and the mobile phase used was: (A) 0.1% formic acid in MilliQ water (pH=2.70) (B) 0.1% formic acid in water: MeCN (10:90), with injection volumes between 4 and 7μL. Samples were prepared in MeOH:MeCN to achieve an approximate concentration of 250ppm.

[0801] The following gradient was used for elution:

[0802]

[0803] Quality parameters

[0804] Probe: ESI capillary

[0805] Source temperature: 120℃

[0806] Probe temperature: 600℃

[0807] Capillary voltage: 0.8KV (+Ve and –Ve)

[0808] Cone voltage: 10 and 30V

[0809] Ionization modes: positive and negative

[0810] LCMS method 2

[0811] LCMS was performed on an Agilent Infinity II G6125C LCMS. The column used was an XBridge C18, 50x4.6 mm, 3.5 μm. The column flow was 1.0 mL / min and the mobile phases used were: (A) 5 mM ammonium bicarbonate in Milli-Q water and (B) MeOH. The injection volume was 5 μL. Samples were prepared in water:MeCN to achieve an approximate concentration of 250 ppm.

[0812] The following gradient was used for elution.

[0813] Time (min) ％A ％B 0.00 92 8 0.75 92 8 3.00 30 70 3.70 5 95 4.20 0 100 5.20 0 100 5.21 92 8 7.00 92 8

[0814] Quality parameters

[0815] Ion source: MMI

[0816] Fragmentation voltage: 70V

[0817] Ionization modes: positive and negative

[0818] Gas temperature: 250℃

[0819] Gasifier: 160℃

[0820] Gas flow rate: 10L / min

[0821] Atomizer pressure: 45psi

[0822] HPLC Method 1

[0823] HPLC analysis was performed on an Agilent Technologies 1100 / 1200 series HPLC system. The column used was ACE 3 C18; 150x4.6mm, 3.0μm particle size (Ex: Hichrom, part number: ACE-111-1546). The flow rate was 1.0mL / min. Mobile phase A was water: trifluoroacetic acid (100:0.1%) and mobile phase B was acetonitrile: trifluoroacetic acid (100:0.1%). The injection volume was 5μL and the following gradient was used:

[0824] Time (min) ％A ％B 0 80 20 35 5 95 39.5 5 95 40 80 20

[0825] Chiral HPLC analysis

[0826] The chiral HPLC data reported were obtained using one of the methods described below.

[0827] Chiral HPLC Method 1

[0828] Chiral HPLC analysis was performed on an Agilent Technologies 1200 Series HPLC system. The column used was CHIRAL PAK IG, 250x4.6 mm, 5 μm. The column flow was 1.0 mL / min and the mobile phase was: (A) 0.1% v / v DEA in n-heptane and (B) IPA:MeOH (70:30). The injection volume was 25 μL. The samples were prepared in IPA:MeOH to achieve an approximate concentration of 250 ppm and the following isocratic method was used:

[0829] time flow ％A ％B 0.01 1.0mL / min 90 10 45 1.0mL / min 90 10

[0830] Chiral HPLC Method 2

[0831] Chiral HPLC analysis was performed on an Agilent Technologies 1200 series HPLC system. The column used was CHIRALPAK IG SFC, 21x250mm, 5μm. The column flow was 1.0mL / min and the mobile phase was: (A) 0.1% v / v DEA in n-heptane and (B) IPA:MeOH (70:30). The injection volume was 20μL. The sample was prepared in IPA:MeOH to achieve an approximate concentration of 250ppm and the following isocratic method was used:

[0832] time flow ％A ％B 0.01 1.0mL / min 85 15 30 1.0mL / min 85 15

[0833] Chiral HPLC Method 3

[0834] Chiral HPLC was performed on an Agilent Technologies 1200 Series HPLC system. The column used was CHIRAL PAK IG, 250x4.6 mm, 5 μm. The column flow was 1.0 mL / min and the mobile phase was: (A) 0.1% v / v DEA in n-heptane and (B) IPA:MEOH (70:30). The injection volume was 10 μL. The sample was prepared in IPA:MeCN to achieve an approximate concentration of 250 ppm and the following isocratic method was used:

[0835] time flow ％A ％B 0.01 1.0mL / min 85 15 25 1.0mL / min 85 15

[0836] Chiral HPLC Method 4

[0837] The conditions were the same as for Chiral Method 3, except the following isocratic method was used:

[0838] time flow ％A ％B 0.01 1.0mL / min 70 30 25 1.0mL / min 70 30

[0839] Chiral HPLC Method 5

[0840] The conditions were the same as for Chiral Method 3, except the following isocratic method was used:

[0841] time flow ％A ％B 0.01 1.0mL / min 90 10 25 1.0mL / min 90 10

[0842] Chiral HPLC Method 7

[0843] Chiral HPLC analysis was performed on an Agilent Technologies 1100 / 1200 Series HPLC system. The column used was CHIRALPAK IA; 250x4.6mm, 5.0μm. The column flow was 1.0mL / min and the mobile phase was: hexane:EtOH:ethanolamine (90:10:0.1%). The injection volume was 5μL. Samples were prepared in 100% EtOH to achieve an approximate concentration of 0.5mg / mL.

[0844] Preparative HPLC method

[0845] Final compounds were purified using one of the following preparative HPLC methods.

[0846] Preparative HPLC Method 1

[0847] Preparative HPLC was performed using a SUNFIRE Prep C18 OBD, 19x250mm, 5μm column with (A) 0.05% HCl in water and (B) 100% MeCN as mobile phases and a flow rate of 17 mL / min and an isocratic system for elution as follows:

[0848] Time (min) flow ％A ％B 00.01 17 70 30 16.00 17 57 43 16.01 17 2 98 18.00 17 2 98 18.01 17 70 30 20.00 17 70 30

[0849] Preparative HPLC Method 2

[0850] Preparative HPLC was performed using an X-bridge prep, C18, 30x250mm, 5μm column with (A) 0.05% HCl in water and (B) 100% MeCN as mobile phases and a flow rate of 25 mL / min with the following isocratic system for elution:

[0851] Time (min) flow ％A ％B 00.01 25 80 20 15.00 25 20 80 15.01 25 2 98 17.00 25 2 98 17.01 25 80 20 19.00 25 80 20

[0852] Preparative Chiral HPLC Method:

[0853] Atropisomers were separated using one of the following preparative chiral HPLC methods.

[0854] Preparative Chiral HPLC Method 1

[0855] Preparative chiral HPLC was performed using a CHIRALPAK IG SFC, 21x250mm, 5μm column with (A) 0.1% DEA in heptane and (B) IPA as mobile phases at a flow rate of 30 mL / min and the following isocratic system:

[0856] Time (min) ％A ％B 0.01 94 6 50.00 94 6

[0857] Preparative Chiral HPLC Method 2

[0858] Preparative chiral HPLC was performed using a CHIRALPAK IG SFC column, 21 x 250 mm, 5 μm, with (A) 0.1% DEA in heptane and (B) IPA:MeOH (90:10) as mobile phases, and a flow rate of 22 mL / min and the following isocratic system for elution:

[0859] Time (min) ％A ％B 0.01 93 7 35.00 93 7

[0860] Chiral Analysis Specific Rotation Solution

[0861] Instruments: Optical Activity AA-10 automatic polarimeter

[0862] Wavelength: 589nm

[0863] Temperature: 23℃

[0864] Pool path length: 1dm

[0865] Solvent: Chloroform (Fisher, HPLC grade)

[0866] Concentration: 1.0g / 100mL

[0867] Sampling technology

[0868] The instrument was switched on and allowed to stabilize for 30 minutes before checking the calibration using the Optical Activity quartz control board (S / N00049). The angular rotation using the sodium yellow D line at 23°C was measured at 34.16° (after first zeroing the instrument without any sample tubes). The sample tube quality was then checked by zeroing the instrument, then filling the sample with chloroform and checking that the instrument still reads 0.00 (+ / - 0.02). The instrument was zeroed in place with a chloroform blank.

[0869] The sample was dissolved in CHCl3 (2 mg in 2 mL), filtered, and 2 mL was pipetted into a cell to measure α.

[0870] The specific rotation is calculated from the following equation: [α]Tλ = (α x 100) / (cl)

[0871] Synthetic intermediates:

[0872] Intermediate A: 1-(4-chlorophenyl)-3-(dimethylamino)propan-1-one hydrochloride

[0873]

[0874] To a solution of 4'-chloroacetophenone (10 g, 65 mmol) in anhydrous EtOH (50 mL) was added paraformaldehyde (1.94 g, 64 mol), N,N-dimethylamine hydrochloride (5.27 g, 64.68 mmol) and concentrated HCl (2 mL) at room temperature. The resulting reaction mixture was stirred at 80-90° C. for 30 h. The reaction mixture was concentrated under reduced pressure and the resulting residue was purified by column chromatography with silica gel (60-120 mesh), eluted with 2% EtOAc / hexane, and triturated with Et2O (100 mL) to provide the title compound (10 g, 40 mmol, 62%).

[0875] Intermediate B: 3-(Dimethylamino)-1-(4-fluorophenyl)propan-1-one hydrochloride

[0876]

[0877] Intermediate B was prepared using the same procedure as described for Intermediate A, except that 4′-fluoroacetophenone (20 g, 144.87 mmol) was used and the resulting residue was purified by column chromatography with silica gel (60-120 mesh), eluting with 4% MeOH / DCM, then triturated with Et2O (400 mL) to provide the title compound (15 g, 77 mmol, 53%).

[0878] Intermediate C: 4-(3-(dimethylamino)propanol)benzonitrile hydrochloride

[0879]

[0880] Intermediate C was prepared using the same procedure as described for Intermediate A, except that 4-acetylbenzonitrile (25 g, 172 mmol) was used and the resulting residue was purified by column chromatography with silica gel (60-120 mesh), eluting with 5% MeOH / DCM, then triturated with Et2O (400 mL) to provide the title compound (20 g, 99 mmol, 57%).

[0881] Example 1

[0882] Preparation of atropisomers A-1 and A-2

[0883] Atropisomers A-1 and A-2 can be prepared by following synthetic route A as shown below.

[0884] Synthetic Route A

[0885]

[0886] Step 1: 4-[4-(4-bromophenyl)-4-oxydeoxybutyryl]benzonitrile

[0887]

[0888] Zinc chloride (30.5g, 223mmol) is heated to melting under vacuum, then cooled to room temperature.Toluene (100mL), tert-butyl alcohol (16.5mL, 172mmol) and TEA (24mL, 172mmol), and the mixture is stirred at room temperature for 2h under nitrogen atmosphere, at this time zinc chloride is completely dissolved.Add 4-cyanoacetophenone (25g, 172mmol) and 4-chlorophenacyl bromide (40.2g, 172mmol), and the reaction mixture is stirred at room temperature for 48h.Reaction mixture is diluted with EtOAc (300mL) and washed with water (5x100mL).The organic extracts merged are dried (Na2SO4) and evaporated under reduced pressure.The residue obtained is purified by grinding with MTBE (400mL), to provide title compound (30g, 101mmol, 59%).

[0889] Step 2: 4-(5-(4-chlorophenyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrrol-2-yl)benzonitrile

[0890]

[0891] A stirred solution of 4-(4-(4-chlorophenyl)-4-oxobutanoyl)benzonitrile (30 g, 101 mmol), 2-trifluoromethylaniline (48.79 g, 303 mmol) and PTSA (1.92 g, 10.099 mmol) in dioxane (300 mL) was heated at 150° C. for 16 h. The reaction mixture was concentrated under reduced pressure and the resulting residue was purified by column chromatography on silica gel (60-120 mesh) using 8% EtOAc / hexanes as eluent to provide the title compound (30 g, 71 mmol, 70%).

[0892] Step 3: 4-(5-(4-chlorophenyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrrol-2-yl)benzoic acid

[0893]

[0894] To a solution of 4-(5-(4-chlorophenyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrrol-2-yl)benzonitrile (2 g, 4.739 mmol) in MeOH (20 mL) was added NaOH (1.89 g, 47 mmol) in water (10 mL), and the resulting mixture was stirred at 90° C. for 24 h. The mixture was concentrated under reduced pressure and the resulting residue was purified by trituration with EtO (10 mL) to provide the title compound (1.8 g, 4.1 mmol, 86%).

[0895] Step 4: 4-(5-(4-chlorophenyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrrol-2-yl)-N-(2-(dimethyl)phenyl (amino)ethyl)benzamide

[0896]

[0897] To a stirred solution of 4-(5-(4-chlorophenyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrrol-2-yl)benzoic acid (1.8 g, 4.0 mmol) in DMF (12 mL) was added DIPEA (2.13 mL, 22 mmol) followed by HATU (4.65 g, 12 mmol). The reaction mixture was stirred at room temperature for 30 min, then N,N′-dimethylethylenediamine (1.08 g, 12 mmol) was added dropwise and stirring was continued for 4 h at room temperature. The mixture was poured into ice-cold water (150 mL) and extracted with EtOAc (3×100 mL). The combined organic layers were dried (Na 2 SO 4 ) and concentrated under reduced pressure. The resulting residue was purified by column chromatography on neutral alumina eluting with 6% MeOH / DCM to provide the title compound (1.2 g, 2.3 mmol, 57%) as a mixture of atropisomers.

[0898] Separation of atropisomers

[0899] Atropisomers of 4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]benzamide (A-1 and A-2) can be resolved by chiral HPLC using preparative chiral HPLC Method 1.

[0900] Two peaks were separated:

[0901] Peak 1: Atropisomer A-1, 4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]benzamide-atropisomer 1 (0.3 g, 0.58 mmol, 38%, >99% ee), and:

[0902] Peak 2: atropisomer A-2, 4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]benzamide-atropisomer 2 (0.31 g, 0.606 mmol, 39%, 98% ee).

[0903] The compound can also be isolated as its hydrochloride salt.

[0904] Example 2

[0905] Further purification and characterization of atropisomers

[0906] Atropisomer A-1: ​​4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2- [(Dimethylamino)ethyl]benzamide hydrochloride

[0907] Peak 1 (0.31 g, 0.606 mmol) was further purified by stirring in HPLC grade water (30 mL), then ultrasonically treated for 10 min and extracted with EtOAc (3x30 mL). The combined organic layer was dried (Na2SO4), filtered and concentrated under reduced pressure, then freeze-dried to provide an amorphous solid (0.290 g, 0.567 mmol, 94%) dissolved in DCM (7.12 mL). The resulting solution was cooled to 0 ° C and the 4N HCl in dioxane (1.42 mL) was added. The reaction mixture was stirred at room temperature for 3 h. The mixture was concentrated and dried under high vacuum. The title compound (0.3 g, 0.56 mmol, 98%) as a pale solid was provided by using Et2O (10 mL) grinding purification and freeze-dried.

[0908] 1 H NMR(DMSO-d6)δ10.03,(brs,1H),8.62(s,1H),7.81-7.68(m,6H),7.25(d,J=8.4Hz,2H), 7.10-7.03(m,4H),6.67-6.58(m,2H),3.56-3.54(m,2H),3.20-3.18(m,2H),2.76(s,6H). LCMS(Method 1)-RT 2.54,MH+512.4

[0909] Atropisomer A-2: 4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2- [(Dimethylamino)ethyl]benzamide hydrochloride

[0910] The hydrochloride salt of atropisomer A-2 was prepared starting from Peak 2 using the same procedure used for atropisomer A-1 to afford the title compound (0.31 g, 0.56 mmol, 99%) as an off-white solid.

[0911] 1H NMR(DMSO-d6)δ9.91(brs,1H),8.69(s,1H),7.81-7.68(m,6H),7.25(d,J=8.0Hz,2H),7 .10-7.03(m,4H),6.67-6.58(m,2H),3.56-3.54(m,2H),3.20-3.18(m,2H),2.77(s,6H). LCMS(Method 1)-RT 2.56,MH+512.4

[0912] Single crystal X-ray crystallographic analysis of atropisomer A-2 (see Example 3 below) indicated that atropisomer A-2 was the R-isomer (compound (1)) and therefore atropisomer A-1 must be the S-isomer.

[0913] Chiral analysis

[0914] Analysis of the chiral properties of atropisomers A-1 and A-2 was performed by measuring their optical rotations and their retention times obtained by chiral HPLC using the method described above to produce the results shown in the table below.

[0915]

[0916] Atropisomer classification

[0917] The isolated atropisomers (atropisomers A-1 and A-2) were subjected to stability studies.

[0918] To evaluate the interconversion of atropisomer A-1 and atropisomer A-2, chiral stability was monitored at 40° C. and 80° C. As shown by the results described below, no interconversion was observed upon heating for 10 days at either temperature.

[0919]

[0920] plan:

[0921] 1. Dissolve 2 x 1 mg of pure atropisomer in 1 mL of EtOH in a sealed dram vial.

[0922] 2. Heat one set of vials at 40°C and the other set at 80°C

[0923] 3. At designated time points, 20 μL aliquots were taken from each stock solution (1 mL) and quenched into HPLC vials containing 80 μL of hexane:EtOH 80:20 solution to provide a final concentration of 200 ppm, and the samples were analyzed by chiral HPLC

[0924] 4. Analyze using chiral HPLC method 5 at the following time points: 0 h, 24 h, 48 h, 72 h, 96 h and 240 h for samples kept at 40 °C, and 24 h, 96 h and 240 h for samples kept at 80 °C

[0925] The stability of the isolated atropisomers (Examples A-1 and A-2) confirmed that they are class 3 atropisomers (LaPlante et al., J. Med. Chem., 54:7005-7022 (2011))).

[0926] Example 3

[0927] X-ray crystallographic analysis of atropisomer A-2

[0928] Atropisomer A-2 free base was prepared and single crystals were subjected to X-ray crystallography as described below.

[0929] experiment:

[0930] Single crystals of atropisomer A-2 with undefined morphology were obtained by recrystallization from methyl isobutyl ketone (MIBK). Select appropriate crystals 0.19 × 0.13 × 0.04 mm 3 , and mounted on a Rigaku XtaLAB Syngery-S diffractometer equipped with a HyPix-6000HE detector using a MiTiGen MicroMount. The crystals were stored at a stable T = 123 (2) K during data collection.

[0931] The data were generated using CuKα radiation. The maximum resolution achieved was θ = 74.263° Data simplification, conversion and absorption correction were performed. The final data integrity was 100.00% at θ 74.263°. The absorption coefficient μ of the compound determined at is 1.761 mm -1 .

[0932] Data were collected and processed using CrysAlisPro software, and the structure was solved using the Intrinsic Phasing solution method with the ShelXT (Sheldrick, 2015) structural solution program and by using Olex2 (Dolomanov et al., 2009) as a graphical interface. The model was refined using ShelXL-2018 / 3 version (Sheldrick, 2018) using least squares minimization.

[0933] The crystal structure was found to be monoclinic and was assigned to space group P21 (#4).

[0934] All non-hydrogen atoms were corrected anisotropically. Hydrogen atom positions were calculated geometrically and corrected using a riding model.

[0935] References: OVDolomanov and LJBourhis and RJGildea andJ.AKHoward and H.Puschmann, Olex2: Acomplete structure solution, refinement and analysis program, J.Appl.Cryst., (2009), 42, 339-341.

[0936] Sheldrick,GM,Crystal structure refinement with ShelXL,Acta Cryst.,(2015),C71,3-8.

[0937] Sheldrick,GM,ShelXT-Integrated space-group and crystal-structuredetermination,Acta Cryst.,(2015),A71,3-8.

[0938] The study results are set forth below in Tables 1-7.

[0939]

[0940]

[0941]

[0942]

[0943]

[0944]

[0945]

[0946]

[0947]

[0948]

[0949]

[0950]

[0951]

[0952]

[0953]

[0954]

[0955] Based on the data described below, it is believed that atropisomer A-2 has Figure 2 and Figure 3 and can therefore be named (R)-4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]benzamide.

[0956] Example 4

[0957] Preparation of atropisomers A-3 and A-4

[0958] Atropisomers A-3 and A-4 were prepared by following synthetic route B as shown below.

[0959] Synthetic route B

[0960]

[0961] Step 1: Diethyl pyridine-2,5-dicarboxylate

[0962] To 2,5-pyridinedicarboxylic acid (20g, 120mmol) in anhydrous EtOH (120mL) suspension in 30min dropwise add concentrated H2SO4 (25.6mL, 0.048mmol). The resulting reaction mixture is refluxed for 48h. The reaction mixture is concentrated, and the resulting residue is alkalized to pH 8 (saturated NaHCO3 aqueous solution). The resulting water layer is extracted with EtOAC (4x200mL). The organic layer is washed with salt water, washed, dried (Na2SO4) and concentrated. Make other four 20g batches of parallel reactions and the resulting crude material from each reaction is merged and purified by column chromatography on silica gel (60-120 mesh) with 5% EtOAC / hexane elution to provide title compound (65g, 291mmol, 49%).

[0963] Step 2: Ethyl 6-(Hydroxymethyl)pyridine-3-carboxylate

[0964] Under nitrogen to pyridine-2,5-diethyl dicarboxylate (10g, 45mmol) in the cooling (ice bath) solution in the mixture of anhydrous EtOH (40mL) and THF (3.5mL) through 30min batches add NaBH4(4.26g, 112mmol) and anhydrous CaCl2(7.86g, 71mmol).The resulting reaction mixture is stirred for 5h at 0 ℃.The reaction mixture is poured into saturated NH4Cl (150mL) aqueous solution and extracted with EtOAc (4x150mL).The organic extracts merged are dried (Na2SO4) and concentrated.Make other six 10g batches and one 5g batches parallel reaction and the gained crude material from each reaction is merged and purified by column chromatography with silica gel (60-120 mesh) eluted with 20% EtOAc / hexane to provide title compound (55g, 320mmol, 100%).

[0965] Step 3: Ethyl 6-formylpyridine-3-carboxylate

[0966] Under nitrogen, DMP (84.32 g, 199 mmol) was added in batches over 20 min to a cooling (ice bath) solution of ethyl 6-(hydroxymethyl)pyridine-3-carboxylate (30 g, 166 mmol) in DCM (360 mL). The reaction was stirred at room temperature for 3 h. The reaction mixture was poured into ice-cold water (1.5 L) and the resulting mixture was alkalized to ~pH 8 (saturated NaHCO aqueous solution) and extracted with EtOAc (4x1000 mL). The combined organic layers were washed with brine, dried (Na2SO4) and concentrated. The resulting residue was purified by column chromatography with silica gel (60-120 mesh), eluted with 12% EtOAc / hexane, to provide the title compound (19 g, 106 mmol, 33%).

[0967] Step 4: 6-[4-(4-chlorophenyl)-4-oxyde-butyryl]pyridine-3-carboxylic acid ethyl ester

[0968] To a stirred solution of intermediate A (1.17 g, 5.6 mmol) and TEA (1.56 mL, 11.2 mmol) in 1,2-dimethoxyethane (10 mL) at room temperature, 6-formylpyridine-3-carboxylic acid ethyl ester (1 g, 5.6 mmol) and 3-ethyl-5-(2-hydroxyethyl)-4-methylthiazole-3-ium bromide (0.28 g, 11.2 mmol) were added. The resulting solution was heated at 80-90 ° C for 5 h. The reaction was diluted with ice-cold water (400 mL) and extracted with EtOAc (3x200 mL). The combined organic layers were dried (Na2SO4) and concentrated. The resulting residue was purified by column chromatography with silica gel (60-120 mesh), eluted with 8% EtOAc / hexane to provide the title compound (5.5 g, 15.9 mmol, 17%).

[0969] Step 5: Ethyl 6-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)phenyl]pyrrol-2-yl]pyridine-3-carboxylate

[0970] To a solution of ethyl 6-[4-(4-chlorophenyl)-4-oxyylidene-butyryl]pyridine-3-carboxylate (2.5 g, 7.2 mmol) in 1,4-dioxane (25 mL) was added 2-aminotrifluorotoluene (3.5 g, 21.7 mmol) and PTSA (0.14 g, 0.72 mmol) at room temperature. The resulting solution was heated at 150° C. for 48 h. The reaction mixture was concentrated and purified by column chromatography with silica gel (60-120 mesh), eluting with 6% EtOAc / hexanes to provide the title compound (2.5 g, 5.3 mmol, 64%).

[0971] Step 6: 6-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)phenyl]pyrrol-2-yl]pyridine-3-carboxylic acid

[0972] To a solution of 6-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)phenyl]pyrrole-2-yl]pyridine-3-carboxylic acid ethyl ester (2.2g, 4.7mmol) in a mixture of THF (10mL) and water (10mL) at room temperature, LiOH (0.59g, 14mmol) was added. The resulting solution was stirred at 80°C for 16h. The reaction mixture was concentrated, diluted with water (150mL) and extracted with EtOAc (4x150mL). The combined organic extracts were dried (Na2SO4) and concentrated. The resulting material was ground with n-pentane (15mL) and Et2O (15mL) to provide the title compound (2g, 4.5mmol, 97%).

[0973] Step 7: 4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethylamino 1-[(-)ethyl]benzamide

[0974] To a solution of 6-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)phenyl]pyrrol-2-yl]pyridine-3-carboxylic acid (2.8 g, 6.33 mol) in DMF (20 mL) was added HATU (7.22 g, 19 mol) and the reaction mixture was stirred at room temperature for 20 min. Unsym-N,N-dimethyl ethylenediamine (1.11 g, 12.7 mol) and DIPEA (3.31 mL, 19 mol) were added and the reaction mixture was stirred at room temperature for 4 h. The reaction mixture was diluted with ice-cold water (200 mL) and extracted with EtOAc (4x100 mL). The combined organic extracts were dried (Na2SO4) and concentrated. The resulting residue was purified by column chromatography with silica gel (60-120 mesh) eluting with 30% EtOAc / hexanes to provide the title compound (2.4 g, 4.7 mmol, 74%).

[0975] Step 8: Separation of atropisomers A-3 and A-4

[0976] The atropisomers of 6-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]pyridine-3-carboxamide can be resolved by chiral HPLC using preparative chiral HPLC method 2.

[0977] Two peaks were separated:

[0978] Peak 1: atropisomer A-3, 6-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]pyridine-3-carboxamide-atropisomer 1 (70 mg, 0.14 mmol, 355%), brown solid.

[0979] Peak 2: Atropisomer A-4, 6-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]pyridine-3-carboxamide-atropisomer 2 (75 mg, 0.15 mmol, 38%), brown solid.

[0980] Both peaks were further purified to remove aliphatic impurities:

[0981] Peak 1: (A-3) (57 mg, 0.11 mmol) was diluted with HPLC grade water (25 mL), then sonicated for 10 min and extracted with EtOAc (3 x 20 mL). The combined organic extracts were dried (Na2SO4), filtered, concentrated and lyophilized to provide atropisomer A-3 (56 mg, 0.11 mmol, 98%, >99% ee).

[0982] 1 H NMR (DMSO-d6) δ8.45-8.43 (m, 2H), 8.01 (d, J = 6.8 Hz, 1H), 7.74-7.68 (m, 2H), 7.65-7.60 (m, 3H), 7.25 (d, J = 8.4 Hz, 2H), 7.11-7.04 (m, 3H), 6.60 (d, J = 4 Hz, 1H), 3.32 (m, 2H, obscured by residual water peak), 2.30 (m, 2H, obscured by residual solvent peak), 2.19 (s, 6H). LCMS (method 1) -RT2.41, MH+513.4

[0983] Peak 2: (A-4): (60 mg, 0.117 mmol) was diluted with HPLC grade water (25 mL), then sonicated for 10 min and extracted with EtOAc (3 x 20 mL). The combined organic extracts were dried (Na2SO4), filtered, concentrated and lyophilized to provide Example A-4 (60 mg, 0.12 mmol, 99%, 95% ee).

[0984] 1 H NMR (DMSO-d6) δ8.47-8.43 (m, 2H), 8.02 (d, J = 7.2 Hz, 1H), 7.74-7.68 (m, 2H), 7.65-7.60 (m, 3H), 7.25 (d, J = 8.4 Hz, 2H), 7.11-7.04 (m, 3H), 6.60 (d, J = 4 Hz, 1H), 3.32 (m, 2H, obscured by residual water peak), 2.30 (m, 2H, obscured by residual solvent peak), 2.20 (s, 6H). LCMS (method 1) -RT2.41, MH+513.4

[0985] Chiral analysis

[0986] Analysis of the chiral properties of atropisomers A-3 and A-4 was performed by measuring their optical rotations and their retention times obtained by chiral HPLC using the method described above to produce the results shown in the table below.

[0987]

[0988] Example 5

[0989] Preparation of atropisomers A-5 and A-6: N-[2-(dimethylamino)ethyl]-6-[5-(4-fluorophenyl)-1-[2- (Trifluoromethyl)phenyl]pyrrol-2-yl]pyridine-3-carboxamide

[0990] Atropisomers A-5 and A-6 were prepared as a racemic mixture using the same procedure as described above for atropisomers A-3 and A-4 in Example 4 with the following exceptions: (a) Intermediate B (3.23 g, 16.58 mmol) was used in Step 4 and 3-benzyl-5-(2-hydroxyethyl)-4-methylthiazol-3-ium bromide (0.678 g, 2.51 mmol) and purified using 10% EtOAc / hexanes as eluent; (b) 1.3% Step 5 purification was performed using EtOAc / hexanes as eluent; (c) MeOH was used instead of THF in step 6 and purification was performed by trituration with Et2O; (d) in step 7, the isolated residue was purified by chromatography on basic alumina gel eluting with DCM to provide the title compound (0.16 g, 0.32 mmol, 55%); (e) purification by preparative HPLC method 1 provided the title compound (61 mg, 0.12 mmol, 38%) (racemic mixture of atropisomers) as its hydrochloride salt (pale yellow solid).

[0991] 1 H NMR(DMSO-d6)δ10.09(bs,1H),8.85(m,1H),8.49(s,1H),8.11(d,J=8.0Hz,1H),7.73–7.70(m,2H),7.69-7.61(m, 3H),7.13–7.10(m,3H),7.06–7.02(m,2H),6.56(d,J=4.0Hz,1H),3.56(m,2H),3.20(m,2H),2.76(d,J=4.4Hz,6H). LCMS (Method 2)-RT 5.06, MH+497.2

[0992] Chiral HPLC analysis using Chiral HPLC Method 3 indicated a mixture of atropisomers, RT peak 1, 9.95 min, 49.8% area (atropisomer A-5) and peak 2, 11.52 min, 50.2% area (atropisomer A-6).

[0993] Example 6

[0994] Preparation of 6-[5-(4-cyanophenyl)-1-[2-(trifluoromethyl)phenyl]pyrrol-2-yl]-N-[2-(dimethylamino Atropisomers A-7 and A-8 of [[(1-(4-(2-(4-(2-(ethyl)pyridine-3-carboxamide)]]]

[0995] Atropisomers A-7 and A-8 were prepared as a racemic mixture using the same procedure as described above for atropisomers A-3 and A-4 in Example 4 with the following exceptions: (a) Intermediate C (0.28 g, 1.39 mmol) was used in step 4 and 3-benzyl-5-(2-hydroxyethyl)-4-methylthiazol-3-ium bromide (0.04 g, 0.14 mmol) and purified using 10% EtOAc / hexanes as eluent; (b) Step 5 purification was performed using 7% EtOAc / hexanes as eluent; (c) In step 7, intermediate C was purified by chromatography on basic alumina gel with 10% The isolated residue was purified eluting with EtOAc / hexane to provide the title compound (0.13 g, 0.25 mmol, 75%); (d) purification by preparative HPLC method 2 provided the title compound (54 mg, 0.11 mmol, 36%) as its hydrochloride salt (pale yellow solid).

[0996] 1 H NMR(DMSO-d6)δ9.83(brs,1H),8.80(t,J=5.2HZ,1H),8.50(d,J=1.6Hz,1H),8.11(dd,J=8.4,2.0Hz,1H),7.79-7. 64(m,6H),7.32-7.07(m,4H),6.83(d,J=4.0Hz,1H),3.56-3.46(m,2H),3.22-3.18(m,2H),2.78(d,J=4.8Hz,6H). LCMS (Method 1)-RT 2.05, MH+504.1

[0997] Chiral HPLC analysis using Chiral HPLC Method 4 indicated a mixture of atropisomers, RT Peak 1, 8.82 min, 50.2% area (Example A-7) and Peak 2, 10.10 min, 49.8% area (Example A-8).

[0998] Example 7

[0999] Preparation of compounds B-2 to B-107

[1000] Additional examples of atropisomer compounds of the invention may be prepared by preparing a racemic mixture of the compounds shown in the following table and then separating the individual atropisomers using the chiral HPLC method described above or a method similar thereto. In the table, the compound numbers given correspond to the example numbers in our earlier international patent application WO2018 / 197714, but with the prefix B- added. Thus, compound B-2 corresponds to Example 2 in WO2018 / 197714, compound B-3 corresponds to Example 3 in WO2018 / 197714, and so on. NMR, LCMS and other characterization data of the racemic compounds and their biological activity data are as given in WO2018 / 197714.

[1001]

[1002]

[1003]

[1004]

[1005]

[1006] Example 8

[1007] Preparation of (R)-4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethyl Alternative method for [amino]ethyl]benzamide (atropisomer A-2)

[1008]

[1009] The title compound was prepared by the following steps 1, 2, 3, 4a and 5a of the synthetic route shown above in Scheme 1. In this route, the chiral resolution was performed on the carboxylic acid intermediate (8) rather than on the dimethylamino-ethylamide (9).

[1010] Step 1: 4-[4-(4-Chlorophenyl)-4-oxydeoxybutyryl]benzonitrile (6)

[1011] Tetrahydrofuran (4mL / g) was loaded into the flask and zinc chloride (1.222g / g, 1.3 equivalents) was added in batches to provide a mobile white suspension, which was stirred for 15min. Tert-butyl alcohol (0.66mL / g, 1 equivalent) was added followed by triethylamine (0.96mL / g, 1 equivalent) in batches, keeping the temperature below 40°C. The reaction was stirred for 2h. 4-cyanoacetophenone (1g / g, 1 equivalent) and 4-chlorophenacyl bromide (1.61g / g, 1 equivalent) were added and the reaction mixture was stirred at 20°C (± 5) for 48h or until the reaction was complete. The product was isolated by precipitation with an aqueous HCl solution and slurry in an aqueous HCl solution and methanol. The resulting solid was dried under vacuum (45°C) to provide the title compound as a light yellow solid.

[1012] Step 2: 4-(5-(4-chlorophenyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrrol-2-yl)benzonitrile (7)

[1013] A flask was charged with 4-(4-(4-chlorophenyl)-4-oxyylidenebutanoyl)benzonitrile (1 g / g, 1 eq) and dioxane (10 mL / g) was added to provide a yellow suspension. 2-Trifluoromethylaniline (1.269 mL / g, 3 eq) was added in a single portion followed by p-toluenesulfonic acid (0.06399 g / g, 0.1 eq) and the reaction mixture was heated at 101°C for 40-72 h (additional portions of p-toluenesulfonic acid (0.1 eq) were added every 8 hours if required to drive the reaction to completion). The reaction mixture was cooled to room temperature and concentrated under vacuum. The resulting oily residue was purified by slurrying in methanol (10 mL / g). The solid was isolated by filtration and dried under vacuum (45°C) to provide the title compound as a yellow solid.

[1014] Step 3: 4-(5-(4-chlorophenyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrrol-2-yl)benzoic acid (8)

[1015] To 4-(5-(4-chlorophenyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrrol-2-yl)benzonitrile (1 g / g, 1 eq) in methanol (10.9 mL / g) was added sodium hydroxide (0.948 g / g, 10 eq) in water (5 mL / g) dropwise over 15 min and the resulting mixture was stirred at 70-76 °C for 18 hours or until completion. The reaction mixture was cooled to room temperature, acidified and the product was isolated by filtration, washed with water (5 mL / g) and acetonitrile (3 mL / g). The product was slurried in acetone / water (20 volumes, 75:25) at 50-55 °C and dried under vacuum (60 °C) to provide the title compound as a yellow solid.

[1016] Step 4a: Chiral separation of (R)4-(5-(4-chlorophenyl)-1-(2-(trifluoromethyl)phenyl)- 1H-pyrrol-2-yl)benzoic acid (3)

[1017] To a flask was added 4-(5-(4-chlorophenyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrrol-2-yl)benzoic acid (1 g / g, 1 eq) followed by tetrahydrofuran (2 mL / g) and acetonitrile (0.75 mL / g). (S)-1-(4-methoxyphenyl)-ethylamine (0.335 mL / g, 1 eq) was added dropwise over 5 min and the resulting reaction mixture was stirred at 40-50 °C for 15 min and then cooled to room temperature. Acetonitrile (7.25 mL / g) was added and the reaction seeded (0.0001 g / g, 99% ee, (S)-1-(4-methoxyphenyl)-ethylamine salt of the desired atropisomer). The reaction mixture was stirred for 16 h and the resulting solid was isolated by filtration, washed with acetonitrile. A hot (75-80° C.) slurry in acetonitrile provided the chiral salt as a white solid (40% yield, 98.16% ee). Salt decomposition was achieved in THF / water (2 / 2 volumes) using 1 M HCl (2.2 equiv.) to provide the acid which was further purified by slurry in water to provide the title compound (90.52 g, 97% salt decomposition yield, 39% overall yield, 98.06% ee). 1 H NMR (DMSO-d6) δ 12,83 (brs, 1H), 7.77-7.67 (m, 6H), 7.23-7.10 (m, 2H), 7.08-7.01 (m, 4H), 6.68 (d, J = 4.0 Hz, 1H), 6.59-6.58 (d, J = 4.0 Hz, 1H). Chiral HPLC using chiral HPLC method 6 showed a single atropisomer, RT 6.083 min, 99.02% area (minor atropisomer RT 7.07 min, 0.98% area).

[1018] Chiral resolution can also be achieved using (S)-(-)-1-phenylethylamine.

[1019] Step 5a: (R)-4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethyl [(1)]-[( ...

[1020] 4-(5-(4-Chlorophenyl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrrol-2-yl)benzoic acid (single atropisomer) (1 g / g, 1 eq) was dissolved in THF (5 mL / g) and N,N-dimethylethylenediamine (0.75 mL / g, 3 eq) was added dropwise followed by DIPEA (1.58 mL / g, 4 eq). 50% T3P in THF (2.72 mL / g, 2 eq) was added dropwise and the reaction mixture was stirred at 20°C for 15 min. Additional portions of 50% T3P in THF were added until the reaction was complete. The reaction mixture was diluted with 10% brine (2 mL / g) and sodium hydroxide solution (2 mL / g) until pH 8-10. The layers were separated and the aqueous layer was extracted with ethyl acetate (2x5 mL / g). The combined organic layers were washed with brine, dried (MgSO4) and concentrated to provide the title compound (80 g, 156 mmol, 71%) as a white triboluminescent solid. Chiral HPLC using Chiral HPLC Method 7 showed a single atropisomer, RT 12.62 min, 99.32% area (minor atropisomer, RT 10.58 min, 0.67% area)

[1021] Example 9

[1022] Preparation and characterization of (R)-4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2- [(Dimethylamino)ethyl]benzamide tartrate

[1023] Method 1: Small-scale preparation of tartrate

[1024] Atropisomer A-2 free base (904.2 mg) was suspended in acetone (9.042 mL, 10 volumes) and stirred for 40 minutes at 25° C. When the solution was free of visible particulates, it was divided into 12 equal aliquots (603 μL), yielding an approximate active content of 60.3 mg per sample.

[1025] An aliquot of 247 μL (1.05 equivalents) of a 0.5 M solution of tartaric acid in ethanol was added to an aliquot of the free base solution at 25° C. The mixture was stirred at 25° C. for 18 hours, after which time a white suspension formed and the resulting solid was then isolated by filtration (PTFE 10 micron sintered cartridge) and the resulting solid was then isolated and dried in vacuo at 40° C. for about 72 hours. The resulting salt was labeled tartrate pattern A (solvate).

[1026] Method 2: Preparation of tartrate salt using isomer A-2 in isopropyl acetate

[1027] Atropisomer A-2 (749.8 mg) was suspended in isopropyl acetate (15 mL, 20 volumes) and the suspension was heated to 40° C. with stirring. When the solution contained no visible particulates, it was divided into 12 equal aliquots (1 ml), yielding an approximate active content of 50 mg per sample. An aliquot of 195.3 μL of a 1 M solution of atropisomer A-2 in ethanol was added to an aliquot of the free base solution at 40° C. The resulting mixture was cooled to 25° C. at a cooling rate of approximately 10° C. / hour. A white suspension formed, and the resulting solid was then isolated by filtration (PTFE 10 micron sintered cylinder) and dried in vacuum at 40° C. for about 18 hours. The resulting salt was labeled as tartrate pattern B.

[1028] Method 3: Preparation of tartrate salt using isopropanol solution of atropisomer A-2

[1029] The tartrate Pattern A salt was prepared by following Method 2, except that atropisomer A-2 (750.1 mg) was initially suspended in isopropanol (15 ml, 20 vol).

[1030] Method 4: Preparation of tartrate salt using a 2-methyl-tetrahydrofuran solution of atropisomer A-2

[1031] Method 1 was repeated, except that atropisomer A-2 (913.9 mg) was initially suspended in 2-methyl-tetrahydrofuran (15 ml, 20 vol), (9.139 mL, 10 vol) and stirred at 25 °C for about 40 min, and then a 250 μl (1.05 eq) aliquot of 1 M tartaric acid in ethanol was added to an aliquot of the A-2 free base solution to produce the tartrate pattern A salt.

[1032] Method 5: 500 mg scale preparation of atropisomer A-2 tartrate pattern B salt

[1033] The atropisomer A-2 free base (521.5 mg) was weighed into a glass vial and charged with isopropyl acetate (20 volumes, 10.430 ml). The mixture was heated to 40°C and stirred for 15 minutes to produce a clear solution. The solution was then charged with tartaric acid (1.05 equivalents, 162.5 mg) dissolved in 3 mL of tetrahydrofuran. The resulting mixture was seeded with atropisomer A-2.tartrate pattern B, which caused the salt to precipitate immediately at 40°C, forming a mobile suspension. The mixture was cooled to 25°C and stirred for 20 hours. The resulting solid was isolated by filtration and dried in vacuo at 40°C to provide the atropisomer A-2 tartrate pattern B salt in 84% yield.

[1034] Method 6: Preparation of Atropisomer A-2 Tartrate Pattern B Salt (Anhydrous Form) by Scale-Up

[1035] Atropisomer A-2 free base (10.0497 g) was weighed into a Buchi flask and charged with isopropyl acetate (20 volumes, 200 ml). The mixture was heated to 40°C to provide a clear solution free of particulates and stirred for 30 minutes. The solution was charged with tartaric acid (3.1954 g, 1.08 equiv) dissolved in tetrahydrofuran (50 mL), the acid was added in batches as follows: 15 mL at 40°C; seeded with atropisomer A-2 tartrate pattern B salt and stirred for 30 minutes; 10 mL and stirred for 1 hour; 10 mL and stirred for 30 minutes; 15 mL and stirred for 30 minutes. The white suspension was then cooled to room temperature at a cooling rate of 10°C / h and stirred for 18 hours. The resulting solid was isolated by filtration in vacuo and washed with isopropyl acetate (2 x 2 volumes) and dried in vacuo at 40 °C for 20 hours to afford A-2 tartrate pattern B salt (anhydrous) in 97% yield; HPLC purity 99.74% (HPLC Method 1), chiral purity 99.27% ​​(Chiral HPLC Method 7).

[1036] Method 7: Preparation of atropisomer A-2 tartrate pattern B salt (anhydrous form) by alternative upscaling by cooling crystallization from butanol / water 96:4

[1037] Atropisomer A-2 free base (36.79 g) was weighed into a flask and charged with butanol (282.57 ml, 7.68 volumes). The mixture was heated to 80°C (light yellow, turbid solution) and stirred for 30 minutes before clarifying into a Mya* vessel preheated at 80°C. The solution was then charged with L-(+)-tartaric acid (1.023 equiv, 11.0806 g) as a solution in water (11.77 mL, 0.32 volumes of initial API charge). The acid solution was clarified by adding dropwise at 80°C. The mixture was then cooled to 68°C over a 30 minute period, seeded with 0.1% ground atropisomer A-2 tartrate pattern B salt seeds (32.6 mg) and held for 1 hour. The mixture was then cooled to 5°C at a cooling rate of 5°C / hour and stirred at 5°C for 6 hours before the solid was isolated. The solid was filtered in vacuo, washed twice with butanol and dried on the filter for 15 minutes, and then dried at 40 °C for 20 hours to provide atropisomer A-2 tartrate pattern B salt (anhydrous) in 83% yield; HPLC purity 99.84% (HPLC method 1), chiral purity 99.66% (chiral HPLC method 7).

[1038] *Note: In the aforementioned equilibration or crystallizations requiring temperature control and / or deterministic heating / cooling profiles, Radley's Mya4 Reaction Station was used. Radley's Mya4 Reaction Station is a 4-zone reaction station with magnetic stirring capability and overhead stirring capability for 2 to 400 mL scale mixtures and a temperature range of -30 to 180°C. The desired reaction conditions were programmed via the Mya 4 control board.

[1039] Characterization of atropisomer A-2 tartrate

[1040] The identity of the salt as a 1:1 (molar ratio free base: tartaric acid) stoichiometric salt is derived from its 1 H NMR spectra were confirmed using a JEOL ECX 400 MHz spectrometer equipped with an autosampler to collect the spectra. Samples were dissolved in appropriate deuterated solvents for analysis. Data were acquired using Delta NMR processing and control software version 4.3.

[1041] The tartrate salt was characterized using X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), gravimetric solubility testing, and gravimetric vapor sorption testing using the techniques described below.

[1042] X-ray powder diffraction (XRPD)

[1043] X-ray powder diffraction patterns were collected on a PANalytical diffractometer using Cu Ka radiation (45 kV, 40 mA), a θ-θ goniometer, a focusing mirror, a divergence slit (1 / 2"), a soller slit (4 mm) at both the incident and divergent beams, and a PIXcel detector. The software used for data collection was X'Pert Data Collector version 2.2f, and the data were presented using X'Pert Data Viewer version 1.2d. XRPD patterns were obtained using a PANalytical X'Pert PRO under ambient conditions with the aid of a transfer foil sample stage (Polyimide-Kapton, 12.7 μm thickness film). The data collection range was 2.994-35° 2θ with a continuous scan speed of 0.202004° s-1.

[1044] Differential Scanning Calorimetry (DSC)

[1045] DSC data were collected on a PerkinElmer Pyris 6000DSC equipped with a 45-position sample holder. Energy and temperature calibrations were verified for the instrument using certified indium. A predetermined amount of sample (0.5-3.0 mg) was placed in an aluminum pan with a pinhole and heated at 20°C.min- from 30°C to 350°C or as determined experimentally. A 20 ml min -1A dry nitrogen purge was maintained. Instrument control, data acquisition, and analysis were performed using Pyris software v11.1.1 revision H.

[1046] Thermogravimetric analysis (TGA)

[1047] TGA data were collected on a PerkinElmer Pyris 1TGA equipped with a 20 position autosampler. The instrument was temperature calibrated using certified weights and certified Alumel and Perkalloy. A predetermined amount (1-5 mg) of sample was loaded onto a pre-tared aluminum crucible and heated at 20 °C.min -1 Heat from ambient temperature to 400°C. Press 20 ml / min above the sample. -1 A nitrogen purge was maintained. Instrument control, data acquisition, and analysis were performed using Pyris software v11.1.1 revision H.

[1048] Weight solubility

[1049] The solubility of salts in water was measured using a gravimetric solubility protocol.

[1050] 1 ml of water was charged to the crystallization tube. The solid was weighed into a tared glass vial, added to the solution in portions and the vial was weighed after each addition until a cloudy solution was observed. The amount in mg was then calculated to yield the solubility in mg / ml.

[1051] The results obtained from the characterization studies are set forth in Table 8 below.

[1052]

[1053]

[1054] Gravimetric Vapor Sorption (GVS)

[1055] GVS studies were performed on atropisomer A-2 tartrate pattern B salt using the protocol described below:

[1056] Adsorption isotherms were obtained using a Hiden Isochema moisture sorption analyzer (model IGAsorp) controlled by IGAsorp system software V6.50.48. The sample was maintained at a constant temperature (25°C) by the instrument control. Humidity was controlled by a mixture of dry and wet nitrogen flows with a total flow rate of 250 ml.min-1. The instrument was verified for relative humidity content by measuring three calibrated Rotronic salt solutions (10-50-88%). The weight change of the sample as a function of humidity was monitored by a microbalance (accuracy + / - 0.005 mg). A defined amount of sample was placed in a tared mesh stainless steel basket under ambient conditions. A complete experimental cycle generally consists of three scans (adsorption, desorption and adsorption) at a constant temperature (25°C) and 10% RH intervals in the range of 0–90% (60 minutes per humidity level). This type of experiment should demonstrate the ability of the sample under study to absorb (or not absorb) moisture over a well-defined set of humidity ranges.

[1057] GVS analysis (see Fig. 9 ) indicates a moisture content of about 0.3% before the first desorption. Between 80% and 90% RH, there is a slightly higher moisture increase, with the solid absorbing about 0.8% moisture. The second absorption / desorption cycle shows how much moisture uptake is fully reversible, returning to 0 wt% at 0% RH. XRPD after a minimum of 3 hours of GVS cycling at 0% RH and 90% RH provides anhydrous pattern B at both RH values.

[1058] It can therefore be concluded that atropisomer A-2 tartrate pattern B salt exists as a stable solid and only absorbs surface water without changing its form.

[1059] Example 10

[1060] Preparation and characterization of (R)-4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2- Other salts of (dimethylamino)ethyl]benzamide

[1061] The hydrochloride, methanesulfonate, maleate, malate, toluenesulfonate, sulfate and phosphate salts of (R)-4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]benzamide have been prepared and characterized. Their X-ray powder diffraction patterns (XRPD), thermal profiles (DSC and TGA) and solubility in water are described in the table below.

[1062] For all salts, 1 H NMR showed a 1:1 ratio between the free base and the counter ion.

[1063]

[1064]

[1065] The solubility of the salt in water was measured using a gravimetric solubility scheme. Therefore, 1 ml of water was loaded into a crystallization tube. The solid was weighed into a tared glass vial, added to the solution in batches and the vial was weighed after each addition until a turbid solution was observed. The amount in mg was then calculated to produce a solubility in mg / mL.

[1066] Proton-NMR

[1067] The data were collected using a JEOL ECX 400 MHz spectrometer equipped with an autosampler. 1 H NMR Spectra. Samples were dissolved in appropriate deuterated solvents for analysis. Data were acquired using Delta NMR Processing and Control Software Version 4.3.

[1068] Preparation of salt

[1069] Example A-2 Small-scale preparation of salt

[1070] Method 1: Acetone-mediated

[1071] Example A-2 free base (904.2 mg) was suspended in acetone (9.042 mL, 10 volumes) and stirred for 40 minutes at 25° C. When the solution was free of visible particulates, it was divided into 12 equal aliquots (603 μL), yielding an approximate active content of 60.3 mg per sample.

[1072] A 0.5 M or 1 M acid stock solution in EtOH (247 μL or 124 μL, 1.05 equiv) was charged to the solution at 25° C. The mixture was stirred for 18 hours at 25° C. If necessary, the sample was further manipulated (e.g., by grinding the solid and adding an antisolvent) to recover the solid for analysis, which was isolated and dried in vacuo at 40° C. for about 72 hours.

[1073] The following table illustrates the amount of acid used, the antisolvent and the resulting crystalline form.Alternative methods can be used to isolate the salts.

[1074]

[1075]

[1076] Method 2: Isopropyl acetate-mediated

[1077] Example A-2 (749.8 mg) was suspended in iPrOAc (15 mL, 20 volumes) heated to 40° C. under agitation. When the solution contained no visible particulates, it was divided into 12 equal aliquots (1 ml), yielding an approximate active content of 50 mg per sample. A 0.5 M or 1 M acid stock solution in EtOH (195.3 μL or 97.7 μL, 1 equivalent) was loaded into the solution at 40° C. The mixture was cooled to 25° C. at approximately 10° C. / h. If necessary, the sample was further manipulated (e.g., by grinding the solid and adding an antisolvent) to recover the solid for analysis, which was isolated and dried in vacuo at 40° C. for approximately 18 h.

[1078] HCl pattern A (TBME antisolvent), tartrate pattern B (1 M acid stock solution in EtOH (195.3 μL)), tosylate pattern A, and phosphate pattern B can be separated by method 2.

[1079] Method 3: IPA-mediated

[1080] The procedure was the same as Method 2, except Example A-2 (750.1 mg) was suspended in IPA (15 mL, 20 vol).

[1081] HCl pattern A (TBME antisolvent), tartrate pattern A (1 M acid stock solution in EtOH (195.3 μL)), tosylate pattern A, and phosphate pattern A can be separated by method 3.

[1082] Method 4: 2-MethylTHF-mediated

[1083] The method was the same as Method 1, except Example A-2 (913.9 mg) was suspended in 2-methylTHF (9.139 mL, 10 vol) and stirred at 25 °C for about 40 min and 0.5 M or 1 M acid stock solution in EtOH (250 μl or 125 μl, 1.05 eq) was used.

[1084] HCl pattern B (heptane as anti-solvent), maleate pattern A (heptane as anti-solvent), tartrate pattern A (1 M acid in EtOH (250 μl, 1.05 eq)) and tosylate pattern A can be separated by method 4.

[1085] Salt subsets are increasingly and more fully characterized.

[1086] Example A-2 500 mg scale preparation of salt

[1087] Hydrochloride

[1088] Example A-2 free base (524.9 mg) was weighed into a glass vial and charged with IPA (20 volumes, 10.498 ml) and heated to 40°C. The solution was stirred at 40°C for 40 min and then charged with HCl (4.4 M in IPA, 1.2 equivalents, 280 μl). The mixture was then seeded with HCl salt pattern B and stirred at 40°C for 15 min before cooling to 25°C. The mixture was concentrated in vacuo to provide a light yellow oil residue. The oil was suspended in 10 volumes of TBME and stirred at 25°C for 72 h to obtain a white suspension. The solid was separated and dried in vacuo at 40°C for 18 h to provide the title salt pattern A in 73% yield.

[1089] Methanesulfonate

[1090] Example A-2 free base (503.9 mg) was weighed into a glass vial and charged with 2-Me THF (10 volumes, 5.039 ml). The mixture was stirred at room temperature for 30 min. The solution was then charged with methanesulfonic acid (1M solution in EtOH, 1.05 equivalents, 1.033 ml), seeded with Example A-2.MsOH pattern A and stirred at 25 ° C for 30 min. The mixture became a turbid solution and then formed a white suspension, which was stirred at 25 ° C for 72 h. The solid was separated by filtration and dried in vacuo at 40 ° C for 18 h to provide the title salt pattern A in a yield of 46%.

[1091] Tartrate

[1092] Example A-2 free base (521.5 mg) was weighed into a glass vial and iPrOAc (20 volumes, 10.430 ml) was loaded. The mixture was heated to 40 ° C and stirred for 15 min to provide a clear solution. The solution was then charged with tartaric acid (1.05 equivalents, 162.5 mg) dissolved in 3 mL of THF. The mixture was then seeded with Example A-2. Tartrate Pattern B, which caused the salt to precipitate immediately at 40 ° C to form a mobile suspension. The mixture was cooled to 25 ° C and stirred for 20 h. The solid was separated by filtration and dried in a vacuum at 40 ° C to provide the title salt Pattern B, with a yield of 84%.

[1093] Tosylate

[1094] Example A-2 free base (504.5 mg) was weighed into a glass vial filled with iPrOAc (20 volumes, 10.090 ml) and heated to 40°C. The solution was stirred at 40°C for 40 min and then p-toluenesulfonic acid (1 M in EtOH, 1.05 eq, 1.04 ml) was charged. The mixture was then seeded with a small amount of Example A-2. Tosylate Salt Pattern A and stirred at 40°C for 15 min before cooling to 25°C. The mixture quickly turned into a white suspension, and it was stirred at 25°C for 72 h. The solid was isolated and dried in vacuo at 40°C for 18 h to provide the title salt Pattern A in 82% yield.

[1095] Maleate

[1096] Example A-2 free base (523.9 mg) was weighed into a glass vial and loaded with 2-Me THF (10 volumes, 5.239 mL). The mixture was stirred at room temperature for 30 min to produce a clear solution. Maleic acid (0.5 M in THF, 1.05 equivalents, 2.149 mL) was then added to the solution, seeded with a small amount of Example A-2. Maleate pattern A and stirred at 25 ° C for 30 min. The mixture was reduced under vacuum to produce a white gum. The gum was suspended in 10 volumes of heptane and stirred at 25 ° C for 72 h. The solid was separated and dried in vacuum at 40 ° C for 18 h to provide the title salt pattern B. 1H NMR was consistent with the structure, but indicated a 1:0.8 stoichiometry.

[1097] Malate

[1098] Example A-2 free base (524.9 mg) was weighed into a glass vial, charged with IPA (20 volumes, 10.618 ml) and heated to 40°C. The solution was stirred at 40°C for 40 min and then malic acid (1M solution in EtOH, 1.05 eq., 1.09 ml) was charged. The mixture was then stirred at 40°C for 15 min before being cooled to 25°C. The mixture, which was kept as a solution at 25°C, was reduced in vacuo, leaving an oily residue. The oil was suspended in 10 volumes of heptane and stirred at 25°C for 70 h to obtain a white suspension. The solid was separated and dried in vacuo at 40°C for 18 h to provide the title salt pattern B.

[1099] Sulfate

[1100] Example A-2 free base (520 mg) was weighed into a glass vial filled with acetone (10 volumes, 5.2 mL). The mixture was stirred at room temperature for 30 min to produce a clear solution.

[1101] The solution was charged with sulfuric acid (1 M in EtOH, 1.05 eq, 1.066 ml), seeded with Example A-2. Sulfate Pattern A and stirred for 30 min at 25° C. The mixture remained as a solution so it was reduced in vacuo with a gentle stream of nitrogen which left a white gum.

[1102] The gum was suspended in 10 volumes of diethyl ether and stirred for 70 h at 25° C. The solid was then isolated and dried in vacuo at 40° C. for 18 h to afford the title salt, Pattern Asim (similar to, but not identical to, the previously isolated sulfate, Pattern A).

[1103] Embodiment 11

[1104] Biological Activity

[1105] A. Assays to measure the effects of compounds of the invention on the viability of U87MG human glioblastoma cancer cells

[1106] The following protocol was used to measure the effects of compounds of the invention on the viability of U87MG cells.

[1107] U87MG cells were grown in their recommended growth medium / supplements (ATCC). Cells were seeded into 96-well plates at 37°C, 5% CO2 overnight at a concentration of 5000 cells / well. Cells were treated with relevant concentrations of test compounds for 72 hours. After 72 hours of incubation, viability was established using a sulforhodamine B (SRB) colorimetric assay. Percent viability was calculated relative to the mean of DMSO-treated control samples, and the IC for inhibition of cell growth was calculated by nonlinear regression (4-parameter logistic equation) using GraphPad Prism software. 50 value.

[1108] Based on the results obtained by following the above protocol, the IC values ​​of the example atropisomers against the U87MG cell line were determined as shown in Table 9 below. 50 value.

[1109]

[1110] * Although individual atropisomers A-5 and A-6 and A-7 and A-8 were identified by chiral chromatography, the racemic mixture was tested in the U87MG cell viability assay.

[1111] B. Assays to measure the effects of atropisomers A-1 and A-2 on cancer cell viability in a diverse panel of cancer cell lines

[1112] Screening for a diversity of cancer cell lines was performed to identify tumor types that exhibit sensitivity to atropisomers A-1 and A-2. A panel of 48 cancer-derived cell lines was screened using dilutions of atropisomers A-1 / A-2 in a high-throughput proliferation assay. Screened cell lines included those representing pancreatic, colorectal / colon, lung, brain, and neurological cancers and lymphoma and leukemia cell lines. Cell lines were treated with half-log serial dilutions of the compounds and proliferation was analyzed 72 hours later using the CellTiter-Glo assay (Promega). IC values ​​were calculated by fitting the dose-response data using a nonlinear regression model. 50 The IC values ​​of atropisomers A-1 and A-2 in micromolar terms are shown in Table 10 below. 50 value.

[1113]

[1114]

[1115] As can be seen from the data, atropisomer A-2 is a significantly more active cytostatic agent than atropisomer A-1 against all cell lines.

[1116] C. Assays to measure the effects of compounds of the invention on cells in mitosis It is known that inhibition of the ability of PLK1 and PLK4 to bind to their partners through their PBDs causes cells to arrest in mitosis. Experimentally, this can be measured by assessing the number of cells in mitosis at a certain time point after treatment with the test compound by immunofluorescence detection of phosphorylated histone H3 (pH3), a marker present only in mitotic cells. PLK1 / 4-PBD inhibitors are expected to cause a dose-dependent increase in pH3-positive cells, reported as the mitotic index (MI) – the percentage of cells positive for this mitotic marker at a given time.

[1117] Different mitotic phenotypes are induced after inhibition of PLK1 and PLK4 in cells. It has been shown that disruption of the PBD domain of PLK1 triggers mitotic arrest with non-dividing chromosomes, a phenotype different from the monopolar spindle phenotype induced by ATP-competitive PLK1 inhibitors (Hanisch et al., 2006 Mol. Biol. Cell 17, 448-459). Centriole assembly is controlled by PLK4, and inhibitors induce multipolar spindle phenotypes due to centrosome defects that produce abnormal cytokinesis (Wong et al., 2015. Science 348 (6239); 1155-1160).

[1118] The following protocol was used to measure the effects of atropisomer A-2 and atropisomer A-3 on cells arrested in mitosis.

[1119] Cells were plated at 10,000 cells / well in 96-well plates and incubated overnight. The next day, a stock solution of atropisomer A-2 in DMSO was diluted in culture medium and then added to the cells at a maximum final concentration of 0.2% DMSO on the cells. Cells were incubated with compounds for 24 hours and then fixed in 3.7% formaldehyde. Cells were permeabilized with 0.1% Triton X-100 and then incubated with anti-phospho-histone H3 (Ser10) antibody (Abcam). Cells were washed with PBS and then incubated with AlexaFluor 488-labeled goat anti-rabbit IgG (Invitrogen) in the presence of 4ug / mL Hoechst 33342 (Invitrogen). Cells were washed in PBS and then imaged using Target Activation V4Bioapplication on an Arrayscan VTi HCS instrument. User-defined thresholds were applied to identify mitotic cells based on the intensity of phospho-histone H3 staining.

[1120] GraphPad Prism was used to plot % mitotic cells against compound concentration using log[inhibitor] versus slope of the response variable with a least squares fit and no constraints.

[1121] From the results obtained by following the above protocol, EC values ​​for HeLa and U87MG cell lines were obtained for atropisomer A-2 and atropisomer A-3. 50 The EC values ​​and the percentage of cells in mitosis are shown in Table 11 below. 50 value.

[1122]

[1123] Phenotypic studies

[1124] In separate studies, for each of atropisomer A-1 and atropisomer A-2, following the above protocol and using a single compound concentration of 0.03 μM, the frequency of mitotic phenotypes observed in U87MG cells was manually assessed and categorized into the following phenotypes for each of A-1 and A-2: non-dividing chromosomes, multipolar spindle / abnormal cytokinesis, monopolar spindle, normal prometaphase, normal metaphase. Fig.10 The results are displayed in .

[1125] result

[1126] Fig.10The results presented in show that atropisomer A-2 has a much greater effect on disrupting normal mitosis than atropisomer A-1. Thus, with A-1, 76% of cells showed a normal mitotic phenotype, comparable to 77% of cells treated with the DMSO control, and 24% of cells showed abnormal cytokinesis compared to 23% of cells treated with the DMSO control. No evidence of a non-dividing chromosome phenotype was seen in cells treated with the DMSO control or atropisomer A-1. In contrast, treatment of cells with the more active atropisomer A-2 resulted in only 17% of cells having a normal mitotic phenotype, 70% of cells having abnormal cytokinesis and 13% of cells having non-dividing chromosomes. These phenotypes are consistent with disrupting PLK1 and PLK4 activity during mitosis.

[1127] D. Assays to measure the effects of atropisomer A-2 on the centrosome

[1128] The results of Study C above show that atropisomer A-2 causes mitotic effects that are characteristic of centrosome dysfunction. Therefore, the effects of A-1 and A-2 on centrosome function were further studied. HeLa cells stably expressing centrosome protein 1-GFP fusion protein were seeded in 96-well plates overnight. The cells were treated with atropisomer A-1 or atropisomer A-2 (concentration 0.02 μM in DMSO) or DMSO control for 72 hours and then imaged using a fluorescence microscope. Multiple cell fields of view were captured for each treatment condition, and the images were then manually analyzed. Centrosome protein 1-GFP specifically marks centrioles as discrete foci and can therefore be used to quantify the number of centrioles per cell. Therefore, for each treatment condition, 100 cells were analyzed and the number of centrioles present in each cell was recorded. The data was then divided into multiple boxes (no centrioles, 1 centriole, 2 centrioles, and more than 2 centrioles) and displayed in Fig.11 middle.

[1129] It can be concluded from the data that atropisomer A-2 shows evidence of a PLK4 inhibitory phenotype in HeLa cells.

[1130] E. Assays to measure the effects of compounds of the invention on the viability of wild-type and KRAS HeLa cells

[1131] A-1, A-2, A-3 and A-4 were tested on HeLa cells designed to inducibly express wild-type or oncogenic KRasG12V transgenes using the FLP-in / T-Rex system (Invitrogen). The cells were plated and then treated with or without doxycycline to induce transgene expression and then treated with serially diluted PBD inhibitors. After 72 hours of incubation, cell viability was assessed using cell titer blue reagent (Promega) and BMG Pherastar microplate reader. The inhibitory effect of PBD on the viability of wild-type or oncogenic G12V KRAS cells was assessed using GraphPad Prism.

[1132] Based on the results obtained by following the above protocol, the GI of each atropisomer against wild-type and KRASG12V HeLa cell lines was determined. 50 Values ​​are shown in Table 12.

[1133]

[1134] F. Kinase Selectivity Assay

[1135] The compounds of the invention bind to the PBD domains of PLK1 and PLK4, but not to the catalytic domains of PLK1 and PLK4, and should show good selectivity over other kinases. Atropisomer A-2 was tested for off-target activity against a panel of ninety-seven kinases distributed across the kinome at a concentration of 3 μM using the DiscoverX KinomeScreen assay. The results are shown in Table 13 below.

[1136] The DiscoverX KinomeScreen assay is a site-directed competitive binding assay that measures the binding affinity of a compound to a kinase by using a solid-supported control compound that can bind or capture kinases in solution. In the absence of a kinase-inhibitor test compound, all kinases will bind to the solid support. If a kinase-inhibitor test compound is added to the assay mixture, the amount of kinase bound to the solid support will be reduced, and the degree of reduction depends on the potency of the test compound as a kinase inhibitor. The potency of the test compound for kinases can be expressed as the percentage of kinases bound to the solid support at a given concentration of the test compound (control percentage), and the lower the percentage, the stronger the kinase binding ability of the test compound. Therefore, a control value percentage of 100% will indicate that the test compound does not bind to the kinase at all, because all kinases are bound to the solid support. Conversely, a control value percentage of 0% will indicate that the test compound has bound to all kinases because no kinases are bound to the solid support.

[1137] plan:

[1138] For most assays, kinase-tagged T7 phage strains were grown in parallel in 24-well blocks in an E. coli host derived from the BL21 strain.

[1139] E. coli was grown to logarithmic phase and infected with T7 phage from frozen stock (infection multiplicity = 0.4) and incubated with shaking at 32°C until cell lysis (90-150 minutes). The lysate was centrifuged (6,000 x g) and filtered (0.2 μm) to remove cell debris. The remaining kinases were produced in HEK-293 cells and subsequently labeled with DNA for qPCR detection. Streptavidin-coated magnetic beads were treated with biotinylated small molecule ligands at room temperature for 30 minutes to produce affinity resins for kinase assays. The ligand-treated beads were blocked with excess biotin and washed with blocking buffer (SeaBlock (Pierce), 1% BSA, 0.05% Tween 20, 1 mM DTT) to remove unbound ligands and reduce nonspecific phage binding. The binding reaction is assembled by merging kinase, ligand-treated affinity beads and test compounds in 1x binding buffer (20% SeaBlock, 0.17x PBS, 0.05% Tween 20, 6mM DTT). The test compound is prepared as a 40x stock solution in 100% DMSO and diluted directly into the assay. All reactions are carried out in a final volume of 0.02ml in a polypropylene 384-well plate. The assay plate is incubated at room temperature for 1 hour under shaking, and the affinity beads are washed with washing buffer (1x PBS, 0.05% Tween 20). The beads are then resuspended in elution buffer (1x PBS, 0.05% Tween 20, 0.5 μM non-biotinylated affinity ligand) and incubated at room temperature for 30 minutes under shaking. The kinase concentration in the eluate is measured by qPCR.

[1140] Compounds that bind to the kinase active site and directly (sterically) or indirectly (allosterically) prevent the kinase from binding to the immobilized ligand will reduce the amount of kinase captured on the solid support. Conversely, test molecules that do not bind to the kinase do not affect the amount of kinase captured on the solid support.

[1141] The strength of binding of the test molecule to the kinase can be expressed as a percentage of the control (% control)

[1142] Percentage of control (% control)

[1143] One or more compounds were screened at 3000 nM concentration and the results of the primary screening binding interactions were reported as '% control', with smaller values ​​indicating stronger hits in the matrix on the following page or pages.

[1144] % Control Calculation

[1145]

[1146] Negative control = DMSO (100% control)

[1147] Positive control = control compound (0% C control)

[1148] The % control values ​​for atropisomer A-2 against the kinase panel are set forth in Table 13 below.

[1149]

[1150]

[1151] Results against ninety-seven kinases showed that atropisomer A-2 had poor or nonexistent binding activity to a broad range of kinases and was therefore unlikely to suffer from problems associated with off-target kinase inhibition.

[1152] In the case of PLK1 and PLK4, atropisomer A-2 showed little or no binding affinity to the catalytic domains of these kinases (% control values ​​were 97% and 100%, respectively). It was therefore concluded that the activity profile indicating PLK1 / PLK4 inhibitory activity demonstrated in the above examples was a result of the non-catalytic polo box domains of PLK1 and PLK4.

[1153] G. Determination of oral bioavailability and brain exposure in mice PK

[1154] Atropisomers A-2 and A-3 were evaluated in an in vivo mouse model to determine brain and plasma concentrations following po and iv administration.

[1155] Follow this protocol:

[1156] Male CD-1 mice were treated with the compounds of Examples A-2 and A-3.

[1157] Administration was by iv administration (2 mg / kg) or by po administration (10 mg / kg).

[1158] Eight samples were taken for analysis in the iv leg at 2, 10, 30 min, 1, 2, 4, 8 and 24 (for iv) and 9 samples were taken in the po leg at 15, 30 min, 1, 2, 4, 8, 24, 48 and 72 h.

[1159] Compounds of Examples A-2 and A-3 were both formulated in 10% DMSO / 90% hydroxypropyl-β-cyclodextrin (20% w / v in water) for iv and po administration. N = 3 mice / time point.

[1160] After administration, peripheral blood samples were taken from each animal and delivered into a labeled polypropylene tube containing an anticoagulant (EDTA). While completing sampling of all animals in the group, the samples were kept on wet ice for up to 30 min. The blood samples were centrifuged to obtain plasma (4°C, 21100g, 5 min) and the resulting plasma was transferred into the corresponding labeled tubes. The terminal brain was cut off from each PO-dosed animal, rinsed with saline and placed in a pre-weighed labeled polypropylene tube, and the sample was weighed again before storage.

[1161] Quantitative bioanalysis was performed using liquid chromatography-mass spectrometry. Tables 14, 15 and 16 below and Figures 12 to 15 The results are displayed in .

[1162] Oral bioavailability

[1163]

[1164]

[1165] The results showed that atropisomers A-2 and A-3 were highly absorbed after oral administration in mice.

[1166] Brain exposure

[1167]

[1168]

[1169] The results of the brain exposure study presented in Table 16 indicate that both atropisomers A-2 and A-3 have high brain exposure. In the case of atropisomer A-2, the results show that atropisomer A-2 has high brain exposure after oral administration in mice with an AUC B:P ratio of 3.3.

[1170] H. In vivo efficacy

[1171] Atropisomer A-2 demonstrated efficacy in a glioblastoma mouse model when tumors were implanted subcutaneously and orthotopically, as shown by the studies described below.

[1172] (i) In vivo anticancer activity in U87MG subcutaneous xenograft model

[1173] Male athymic nude mice bearing U87MG tumors were given oral doses of atropisomer A-2 at 100 mg / kg on days 1, 4, and 7 and tumor volume was measured for 20 days. Tumor volume was also measured in a control group of tumor-bearing mice that received vehicle only at the same time points. The treated group showed a significant reduction in tumor volume compared to the control (3.85% T / C on day 13), as shown in Table 1. Fig.16 as shown in .

[1174] (ii) In vivo anticancer activity of U87-Luc in an orthotopic xenograft model

[1175] U87-Luc cells were implanted intracerebrally into the brains of male athymic nude mice and tumor growth was monitored by bioluminescent signals. In the treatment groups, animals were given a 100 mg / kg oral dose of atropisomer A-2 on days 1, 4, 7, 10, and 13. Control animals were given vehicle only. Fig.17 The results shown in Figure 4 indicate that at day 15, the tumor signal in the treated group decreased compared to the control group.

[1176] (iii) In vivo anticancer activity in HCT116 tumor-bearing mice

[1177] Atropisomer A-2 has shown efficacy in KRAS mutant colorectal cancer models, as described below.

[1178] Male athymic nude mice bearing HCT116 xenograft tumors were given a 100 mg / kg oral dose of atropisomer A-2 and tumor volume was measured for 3 weeks on days 1, 8, and 15. Tumor volume was also measured in a control group of tumor-bearing mice that received vehicle only at the same time points.

[1179] Fig.18 The results shown in show that ® significantly affected tumor growth at day 20 (TGI 60%).

[1180] Pharmaceutical preparations

[1181] (i) Tablet preparation

[1182] A tablet composition comprising the composition of matter or atropisomer of the present invention is prepared by mixing 50 mg of the compound with 197 mg of lactose (BP) as a diluent and 3 mg of magnesium stearate as a lubricant and compressing in a known manner to form tablets.

[1183] (ii) Capsule preparation

[1184] Capsule formulations are prepared by mixing 100 mg of the composition of matter or atropisomer of the invention with 100 mg of lactose and filling the resulting mixture into standard opaque hard gelatin capsules.

[1185] (iii) Injection Preparation I

[1186] Parenteral compositions for injection can be prepared by dissolving the composition of matter or atropisomers (eg, in salt form) of the invention in water containing 10% propylene glycol to give a concentration of 1.5% by weight of active compound. The solution is then sterilized by filtration, filled into ampoules and sealed.

[1187] (iv) Injection preparation II

[1188] The parenteral composition for injection is prepared by dissolving the composition of matter or atropisomer (eg, in salt form) of the invention (2 mg / ml) and mannitol (50 mg / ml) in water, sterile filtering the solution and filling sealable 1 ml vials or ampoules.

[1189] (v) Injection preparation III

[1190] A formulation for delivery by injection or infusion iv can be prepared by dissolving the composition of matter or atropisomer of the invention (eg, in salt form) at 20 mg / ml in water. The vial is then sealed and sterilized by autoclaving.

[1191] (vi) Injection preparation IV

[1192] Formulations for delivery by injection or infusion iv can be prepared by dissolving the composition of matter or atropisomer of the invention (eg, in salt form) at 20 mg / ml in water containing a buffer (eg, 0.2 M acetate pH 4.6). The vial is then sealed and sterilized by autoclaving.

[1193] (vii) Subcutaneous injection preparations

[1194] A composition for subcutaneous administration is prepared by mixing the composition of matter or atropisomer of the invention with pharmaceutical grade corn oil to give a concentration of 5 mg / ml. The composition is sterilized and filled into suitable containers.

[1195] (viii) Lyophilized preparations

[1196] Aliquots of the formulated composition of matter or atropisomer of the invention were placed in 50 ml vials and lyophilized. During lyophilization, the composition was frozen at (-45°C) using a one-step freezing protocol. The temperature was raised to -10°C for annealing, then lowered to -45°C for freezing, followed by primary drying at +25°C for about 3400 minutes, then if the temperature reached 50°C, a step was added for secondary drying. The pressure during primary and secondary drying was set to 80 mTorr.

[1197] Equivalent

[1198] The foregoing examples are presented for the purpose of illustrating the present invention and should not be construed as imposing any limitation on the scope of the present invention. It is apparent that many modifications and substitutions may be made to the specific embodiments of the present invention described above without departing from the underlying principles of the present invention and are described in the examples. All such modifications and substitutions are intended to be encompassed in the present application.

Claims

1. A composition of matter, consisting of: (i) at least 90% by weight of the atropisomer of formula (1) or a pharmaceutically acceptable salt thereof; and (ii) 0-10% by weight of an atropisomer of formula (2) or a pharmaceutically acceptable salt thereof.

2. A single atropisomer having formula (1) The single atropisomer is not accompanied by any other atropisomer, or is accompanied by no more than 0.5% by weight of any other atropisomer relative to the single atropisomer.

3. A single atropisomer (R)-4-[5-(4-chlorophenyl)-1-[2-(trifluoromethyl)-phenyl]pyrrol-2-yl]-N-[2-(dimethylamino)ethyl]benzamide or a pharmaceutically acceptable salt thereof.

4. A single atropisomer of the formula having an optical rotation value less than 0:

5. A pharmaceutical composition comprising the composition of matter or a single atropisomer according to any one of claims 1 to 4 and a pharmaceutically acceptable excipient.

6. The pharmaceutical composition according to claim 5, further comprising one or more other therapeutic agents.

7. Use of the composition of matter or a single atropisomer according to any one of claims 1 to 4 in the manufacture of a medicament for treating cancer.

8. The use according to claim 7, wherein the cancer is selected from: a) pancreatic cancer, colon and colorectal cancer, lung cancer, breast cancer, ovarian cancer, prostate cancer, and brain and nerve cancer, b) Blood cancer, and myelodysplastic syndrome, c) glioma and glioblastoma, d) Rhabdoid tumor; medulloblastoma and other embryonal brain tumors; melanoma, gastric cancer, e) sarcoma of soft tissue, bone or cartilage, f) Cancers involving PLK1, g) Cancers involving PLK4, h) cancers characterized by p53 deficiency or mutations in the TP53 gene, i) Cancers characterized by the presence of mutant forms of KRAS.

9. The use according to claim 7, wherein the cancer is selected from the group consisting of lymphoma, leukemia, glioblastoma multiforme, ependymoma, diffuse intrinsic pontine glioma, IDH1 mutant glioma, rhabdomyosarcoma, osteosarcoma, PLK1 overexpressed cancer, PLK4 overexpressed cancer.

10. The use according to claim 7, wherein the cancer is selected from the group consisting of: AML, ALL, B and T cell lymphoma.

11. A method for preparing a composition of matter or a single atropisomer according to any one of claims 1 to 4, the method comprising chiral separation of a mixture of atropisomers of a compound of formula (0):

12. The method of claim 11, wherein the mixture of atropisomers of the compound of formula (0) is a racemic mixture.

13. The method of claim 11, wherein the chiral separation is performed by: (i) passing the mixture of atropisomers through a chiral chromatography column; or (ii) reacting a mixture of atropisomers of the compound of formula (0) with a chiral acid to form salts of two atropisomers in the mixture, separating the salts and decomposing the salts to produce the corresponding free base of each atropisomer.

14. The method of claim 13, wherein the chiral chromatography column is a chiral HPLC column.

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