CA3132995A1 - Compositions and methods for treating androgen receptor positive forms of cancer - Google Patents

Abstract

Pharmaceutical compositions comprising a CBP Inhibitor compound can be used to treat patients diagnosed with Androgen Receptor positive forms of cancer, such as mCRPC and TNBC, including patients diagnosed with the AR-v7 splice form.

Description

2 PCT/US2020/022823 COMPOSITIONS AND METHODS FOR TREATING
ANDROGEN RECEPTOR POSITIVE FORMS OF CANCER
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims the benefit of U.S. Provisional Application No.
62/819,487, filed March 15, 2019, U.S. Provisional Application No. 62/819,482, filed March 15, 2019, U.S.
Provisional Application No. 62/819,472, filed March 15, 2019, U.S. Provisional Application No.
62/819,490, filed March 15, 2019, U.S. Provisional Application No. 62/819,476, filed March 15, 2019, U.S. Provisional Application No. 62/821,660, filed March 21, 2019, and International Application No. PCT/U52019/039936, filed June 28, 2019, each of which is incorporated by reference herein in its entirety.
TECHNICAL FIELD
[0002] This disclosure relates to compositions and methods for inhibiting CREB-binding protein (CBP). Compositions for inhibiting CBP are useful, for example, in pharmaceutical compositions for the treatment of certain Androgen Receptor dependent forms of cancer.
BACKGROUND

[0003] Growth and proliferation of hormone sensitive tumors are dependent on oncogenic signaling programs driven by corresponding nuclear hormone receptors. The androgen receptor (AR), a key driver in prostate cancer and subsets of breast cancers, controls the expression of about 100 androgen-responsive target genes. Expression of these AR target genes is important for normal tissue development and cellular activities but can have pathological effects that underlie tumor initiation and progression. Direct targeting of androgen biosynthesis and androgen interaction with the AR can provide clinical utility. However, acquired resistance to these therapies can circumvent ligand-driven AR function while retaining continued dependence on AR-driven transcriptional programs.

[0004] Nuclear receptors are part of multiprotein complexes involving co-activators and co-repressors which control the impact of the nuclear receptor on its downstream target genes.
Within the AR-associated multiprotein complex, CBP/P300 are critical co-activators of AR, modifying the chromatin environment surrounding the nuclear receptor to increase its intrinsic transcriptional activity and recruiting additional co-factors. Given the co-regulatory relationship between the AR and CBP/P300, inhibition of CBP/P300 activity offers a rational approach to suppress AR-dependent oncogenic programs in AR-dependent tumors such as breast and prostate cancers.

[0005] There are no approved therapeutics specifically aimed at metastatic castrate-resistant (mCRPC) patients for whom androgen antagonists and taxane therapies have proven ineffective.
These include patients with tumors harboring structurally altered androgen receptors, including the AR-v7 splice form, which continue to promote the AR transcriptional program in a ligand-independent manner, unaffected by androgen antagonists. This population represents an unmet clinical need.
SUMMARY

[0006] The present invention provides methods of and compositions for treating AR+ cancers, including patients diagnosed with certain forms of AR+ cancer that are resistant to other treatments, such as patients resistant or refractory to apalutamide, darolutamide or enzalutamide (e.g., patients with disease progression or with disease refractory to treatment with enzalutamide), by the administration of a CBP Inhibitor compound (e.g., compounds of formula (I)) to a patient in need thereof. The CBP Inhibitor compositions are preferably used in a therapeutically effective amount to inhibit CBP and antagonize androgen receptor signaling, leading to clinical benefit in AR+ Triple Negative Breast Cancer (TNBC), and mCRPC expressing the AR-v7 splice form. AR
is an oncogenic driver in prostate cancer and progression of the disease towards castration- and drug-resistance is associated with aberrations of AR such as amplification of AR, mutations in the LBD and increase in a splice variant of AR lacking the LBD (AR-v7). AR is also expressed in a subset of TNBC in which it substitutes for ER and drives ER signaling via binding to ER response elements. Finally, ER is the main oncogenic driver in ER+ breast cancer.
Resistance to hormone deprivation in this subtype of breast cancer can lead to mutations in ER LBD
leading to ligand-independent growth. In one aspect, the CBP Inhibitor is useful to treat hormone-receptor positive subjects with relapsed or refractory-to-hormone-therapy cancers (e.g., by antagonizing ER activity in these subjects).

[0007] The present disclosure is based in part on the discovery that compounds of formula (I):

(R2)11 \ /

N ¨
N.----9..
N OH

0 0 (I) and pharmaceutically acceptable salts thereof, wherein:
R1 is H or ¨OH;
each R2 is independently selected from C1-C6 alkyl (e.g., methyl), halogen, ¨CN, and ¨0R3, (e.g., methoxy) wherein the alkyl is optionally substituted with one or more halogen;
each R3 is independently H or Ci-C6 alkyl (e.g., methyl), wherein the alkyl is optionally substituted with one or more halogen; and n is an integer selected from 0, 1, 2, 3, 4 or 5, wherein n is preferably 0, 1, 2, or 3, provide an active moiety useful for the treatment of AR-positive cancer, such as certain AR-positive forms of breast cancer (e.g., TNBC) and prostate cancer (e.g., CRPC).

[0008] The disclosure includes the use of compounds of formula (I), and pharmaceutically acceptable salts thereof, for the treatment of diseases or disorders associated with the inhibition of CBP, including certain AR-positive forms of cancer, including the AR-v7 splice form of AR. As an essential cofactor in AR-driven transcription, including that of androgen-independent AR
variants, CBP/P300 is an attractive target for development of novel therapy to meet these patients' needs.

[0009] In some embodiments, the compound of formula (I) is Compound 1:
F . 0/
OH
N-Nh.9....
N

I (1) or a stereoisomer and/or pharmaceutically acceptable salt thereof.

[0010] In some embodiments, Compound 1 is the first eluting isomer when eluted by preparative HPLC under the conditions defined in Example 1.2.

[0011] In some embodiments, the compound of formula (I) is Compound 2:
OH
N-(2) or a stereoisomer and/or pharmaceutically acceptable salt thereof.

[0012] In some embodiments, Compound 2 is the first eluting isomer when eluted by preparative HPLC under the conditions defined in Example 1.3.

[0013] In some embodiments, the compound of formula (I) is Compound 3:
F 4.0 0>-F
OH
N-0 o (3) or a stereoisomer and/or pharmaceutically acceptable salt thereof.

[0014] In some embodiments, Compound 3 is the second eluting isomer when eluted by preparative HPLC under the conditions defined in Example 1.4.

[0015] In some embodiments, the compound of formula (I) is Compound 4:

.,,OH

(4) or a stereoisomer and/or pharmaceutically acceptable salt thereof.

[0016] Compounds of formula (I) are active in enzalutamide-resistant preclinical models, and thus represent a potential novel therapy for these patients with refractory or resistant disease.
BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 is a table of compounds in accordance with various embodiments of the disclosure.

[0018] Figure 2 is a series of reaction schemes for chemical syntheses of compounds of Formula (I) and useful intermediates in the preparation of compounds of Formula (I).

[0019] Figure 3 is an immunoblot showing H3K27Ac, total H3, and 13-actin protein expression levels from a breast cancer cell line exposed to increasing concentrations of compound 1 for 24h.

[0020] Figure 4 is a graph showing the in vivo activity of Compound 1 in a cell line derived xenograft model of AR+ triple negative breast cancer.

[0021] Figure 5 is an immunoblot showing AR and AR-v7 expression levels in an AR-v7+ prostate cancer cell line after 24h exposure to increasing concentrations of Compound 3.

[0022] Figure 6 is a graph showing the in vivo activity of Compound 4 in a patient-derived xenograft model of prostate cancer resistant to enzalutamide.
DETAILED DESCRIPTION

[0023] The present disclosure encompasses the recognition that compounds of formula (I) are CBP Inhibitor Compounds, defined herein as compounds having one or more of the following characteristics when tested according to the HTRF biochemical Assay Protocol below in Example 2: (1) a CBP IC50 value of less than 1 (1M; and (2) a CBP IC50 value of between 0.001 and 1 (1M.
CBP Inhibitor Compounds can be of formula (I):
(12_ )n \ /
Ri N¨

N----9._ N OH

00 (I) or a pharmaceutically acceptable salt thereof, wherein R1, R2, and n are as described above.

[0024] Unless otherwise indicated herein, all isomeric forms of specified chemical compounds are provided by the present disclosure, including mixtures thereof (e.g., S, R and racemic orientations at each chiral center). If the compound contains a double bond, the substituent may be in the E or Z configuration. If the compound contains a disubstituted cycloalkyl, the cycloalkyl substituent may have a cis- or trans- configuration. All tautomeric forms are also intended to be included.

[0025] Compounds of formula (I) and Group A, unless otherwise indicated, may exist in their tautomeric form. All such tautomeric forms are contemplated herein as part of the present disclosure.

[0026] The compounds of formula (I) and Group A, unless otherwise indicated, may contain one or more stereocenters, and, therefore, exist in different stereoisomeric forms. It is intended that unless otherwise indicated all stereoisomeric forms of the compounds of formula (I) and Group A, as well as mixtures thereof, including racemic mixtures, form part of the present disclosure. In addition, the present disclosure embraces all geometric and positional isomers. For example, if a compound of formula (I) or Group A incorporates a double bond or a fused ring, both the cis- and trans-forms, as well as mixtures, are embraced within the scope of the disclosure. Each compound herein disclosed includes all the enantiomers that conform to the general structure of the compound. The compounds may be in a racemic or enantiomerically pure form, or any other form in terms of stereochemistry. The assay results may reflect the data collected for the racemic form, the enantiomerically pure form, or any other form in terms of stereochemistry.

[0027] Diastereomeric mixtures can be separated into their individual diastereomers based on their physical chemical differences by methods well known to those skilled in the art, such as, for example, by chromatography and/or fractional crystallization. Enantiomers can be separated by converting the enantiomeric mixture into a diastereomeric mixture by reaction with an appropriate optically active compound (e.g., chiral auxiliary such as a chiral alcohol or Mosher's acid chloride), separating the diastereomers and converting (e.g., hydrolyzing) the individual diastereomers to the corresponding pure enantiomers. Also, some of the compounds of formula (I) or Group A may be atropisomers (e.g., substituted biaryls) and are considered as part of this disclosure. Enantiomers can also be separated by use of a chiral HPLC column.

[0028] The compounds of formula (I) or Group A may form acid addition salts, which may be pharmaceutically acceptable salts. The disclosure also includes pharmaceutical compositions comprising one or more compounds as described herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. In some embodiments, pharmaceutical compositions reported herein can be provided in a unit dosage form (e.g., capsule, tablet or the like). In some embodiments, pharmaceutical compositions reported herein can be provided in an oral dosage form. In some embodiments, an oral dosage form of a compound of formula (I) or Group A can be a capsule. In some embodiments, an oral dosage form of a compound of formula (I) or Group A is a tablet. In some embodiments, an oral dosage form comprises one or more fillers, disintigrants, lubricants, glidants, anti-adherents and/or anti-statics. In some embodiments, an oral dosage form is prepared via dry blending. In some embodiments, an oral dosage form is a tablet and is prepared via dry granulation.

[0029] A CBP Inhibitor compound of the present disclosure can be dosed at a therapeutically effective level. A Selective CBP Inhibitor compound of the present disclosure can be dosed at a therapeutically effective level.
Compounds of the Disclosure

[0030] In one aspect, the disclosure relates to compounds of formula (I):
(12_ )n \ /
Ri N-N----9._ N OH

00 (I) or a pharmaceutically acceptable salt thereof, wherein:
R1 is H or ¨OH;
each R2 is independently selected from C1-C6 alkyl, halogen, ¨CN, and ¨0R3, wherein the alkyl is optionally substituted with one or more halogen;
each R3 is independently H or Ci-C6 alkyl, wherein the alkyl is optionally substituted with one or more halogen; and n is an integer selected from 0-5, wherein n is preferably 0, 1, 2, or, 3.

[0031] In some embodiments, R1 is H or ¨OH; each R2 is independently selected from ¨F, ¨Cl, ¨CH3, ¨CHF2, ¨CN, and ¨0R3; each R3 is independently selected from ¨CH3, ¨CHF2, and ¨CH(CH3)2, and n is selected from 0, 1, 2, and 3.

[0032] In some embodiments, compounds of Formula (I) are provided wherein R2 is ¨Cl, ¨CH3, ¨CHF2, ¨CN, ¨OCH3, ¨OCHF2, ¨OCH(CH3)2. In some embodiments, R2 is ¨F, ¨CH3, ¨CHF2, ¨CN, or ¨0R3. In some embodiments, R2 is ¨F, ¨Cl, ¨CHF2, ¨CN, or ¨0R3. In some embodiments, R2 is ¨F, ¨Cl, ¨CH3, ¨CN, or ¨0R3. In some embodiments, R2 is ¨F, ¨Cl, ¨CH3, ¨CHF2, or ¨0R3.
In some embodiments, R2 is ¨F, ¨Cl, ¨CH3, ¨CHF2, or ¨CN.

[0033] In one embodiment, the disclosure provides compounds of formula (lb):
R2c R2b R2d R2a R
R2e 1 N¨

N
OH

0 0 (Ib) or a pharmaceutically acceptable salt thereof, wherein:
R1 is H or ¨OH;
R2a is selected from H, C1-C6 alkyl (e.g., methyl), halogen (e.g., F or Cl), and ¨0R3, wherein the alkyl is optionally substituted with one or more halogen (e.g., CHF2);
R2b is selected from H, C1-C6 alkyl (e.g., methyl), halogen (e.g., F or Cl), and ¨0R3, wherein the alkyl is optionally substituted with one or more halogen (e.g., CHF2);
R2' is selected from H, C1-C6 alkyl (e.g., methyl), halogen (e.g., F or Cl), and ¨0R3, wherein the alkyl is optionally substituted with one or more halogen;
R2d is selected from H or halogen (e.g., F or Cl);
R2e is selected from H, C1-C6 alkyl (e.g., methyl), halogen (e.g., F or Cl), ¨CN, and ¨0R3, wherein the alkyl is optionally substituted with one or more halogen (e.g., methyl); and R3 is H or C1-C6 alkyl (e.g., methyl), wherein the alkyl is optionally substituted with one or more halogen (e.g., CHF2).

[0034] In one embodiment, the disclosure provides compounds of formula (lb):

R2c R2b R2d R2a R
R2e 1 N¨

N
OH

0 0 (Ib) or a pharmaceutically acceptable salt thereof, wherein:
R1 is ¨OH;
R2a is selected from H, Ci-C6 alkyl (e.g., methyl), halogen (e.g., F or Cl), and ¨0R3, wherein the alkyl is optionally substituted with one or more halogen (e.g., CHF2);
R2b is selected from H, Ci-C6 alkyl (e.g., methyl), halogen (e.g., F or Cl), and ¨0R3, wherein the alkyl is optionally substituted with one or more halogen (e.g., CHF2);
R2' is selected from H, Ci-C6 alkyl (e.g., methyl), halogen (e.g., F or Cl), and ¨0R3, wherein the alkyl is optionally substituted with one or more halogen;
R2d is selected from H or halogen (e.g., F or Cl);
R2e is selected from H, C1-C6 alkyl (e.g., methyl), halogen (e.g., F or Cl), ¨CN, and ¨0R3, wherein the alkyl is optionally substituted with one or more halogen (e.g., methyl); and R3 is H or C1-C6 alkyl (e.g., methyl), wherein the alkyl is optionally substituted with one or more halogen (e.g., CHF2).

[0035] In one embodiment, the disclosure provides compounds of formula (lb):
R2c R2b R2d R2a R
R2e 1 N¨

N
OH

0 0 (Ib) or a pharmaceutically acceptable salt thereof, wherein:

R1 is H;
R2a is selected from H, Ci-C6 alkyl (e.g., methyl), halogen (e.g., F or Cl), and ¨0R3, wherein the alkyl is optionally substituted with one or more halogen (e.g., CHF2);
R2b is selected from H, Ci-C6 alkyl (e.g., methyl), halogen (e.g., F or Cl), and ¨0R3, wherein the alkyl is optionally substituted with one or more halogen (e.g., CHF2);
R2' is selected from H, Ci-C6 alkyl (e.g., methyl), halogen (e.g., F or Cl), and ¨0R3, wherein the alkyl is optionally substituted with one or more halogen;
R2d is selected from H or halogen (e.g., F or Cl);
R2e is selected from H, C1-C6 alkyl (e.g., methyl), halogen (e.g., F or Cl), ¨CN, and ¨0R3, wherein the alkyl is optionally substituted with one or more halogen (e.g., methyl); and R3 is H or C1-C6 alkyl (e.g., methyl), wherein the alkyl is optionally substituted with one or more halogen (e.g., CHF2).

[0036] In one embodiment, the disclosure provides compounds of formula (lb):
R2c R2b R2d R2a R
R2e 1 N¨

N
OH

(Ib) or a pharmaceutically acceptable salt thereof, wherein:
R1 is H or -OH;
R2a is selected from H, C1-C6 alkyl (e.g., methyl), halogen (e.g., F or Cl), and ¨0R3, wherein the alkyl is optionally substituted with one or more halogen (e.g., CHF2);
R2b is H;
R2' is H;
R2d is independently selected from H or halogen (e.g., F or Cl);
R2e is H; and R3 is independently H or C1-C6 alkyl (e.g., methyl), wherein the alkyl is optionally substituted with one or more halogen (e.g., CHF2).

[0037] In one embodiment, the disclosure provides compounds of formula (lb):

R2c R2b R2d R2a R
R2e 1 N¨

N
OH

0 0 (Ib) or a pharmaceutically acceptable salt thereof, wherein:
R1 is H or -OH;
R2a is selected from H, and ¨0R3;
R2b is H;
R2' is H;
R2d is independently selected from H or halogen (e.g., F or Cl);
R2e is H; and R3 is independently H or Ci-C6 alkyl (e.g., methyl), wherein the alkyl is optionally substituted with one or more halogen (e.g., CHF2).

[0038] In some embodiments, Selective CBP Inhibitor Compounds of formula (I) are provided.
In some embodiments, Selective CBP Inhibitor Compounds of Formula (lb) are provided. The present disclosure encompasses the recognition that compounds of formula (I) are CBP Inhibitor Compounds, defined herein as compounds having one or more of the following characteristics when tested according to the HTRF biochemical Assay Protocol below in Example 2: (1) a CBP
IC50 value of less than 1 i.tM; and (2) a CBP IC50 value of between 0.001 and 1 i.i.M.

[0039] In some embodiments, the disclosure relates to compounds of formula (I) that are of a formula selected from Group A:

(R2), (R2)õ
(1.._....... 1 e \
\ 3.
OH OH
N-Ni....( ?... Ni,,Q
:.
0 i.
N N )--OH OH

I (Al), I (A2), (R2) (R2) ,, ,, .,,0,_, ='10H
N- N-0 i/.
N

i....9.. Ni...0 N N -OH OH

1 (A3), I (A4), (r-2 (R2),, OH '10H
,.9., Nh N-Ni .9., N N
OH OH

1 (A5), I (A6), (R2),, (R2L
\ 31 (7____..1 e \
OH .'10H
N-N...0 :.
N N

1 (A7), I (A8), (R2) (R2),, N-(A9), I (A10), (R2) (R2),, N¨ N-(All), and I (Al2), and pharmaceutically acceptable salts thereof.

[0040] In some embodiments, the disclosure relates to a compound of formula (I) selected from Figure 1. In Figure 1, "Eluted Isomer" refers to the order in which the compound eluted by preparative HPLC.

[0041] In some embodiments, R1 is H or ¨OH. In some embodiments, R1 is H. In some embodiments, R1 is ¨OH.

[0042] In some embodiments, each R2 is independently selected from C1-C6 alkyl, halogen, ¨CN, and ¨0R3, wherein the alkyl is optionally substituted with one or more halogen. In some embodiments, R2 is Ci-C6 alkyl, wherein the alkyl is optionally substituted with one or more halogen. In some embodiments, R2 is Ci-C6 alkyl, wherein the alkyl is substituted with one halogen. In some embodiments, R2 is Ci-C6 alkyl, wherein the alkyl is substituted with two halogens. In some embodiments, R2 is selected from ¨CH3 and ¨CHF2. In some embodiments, R2 is ¨CH3. In some embodiments, R2 is ¨CHF2. In some embodiments R2 is halogen.
In some embodiments, R2 is selected from ¨F and ¨Cl. In some embodiments, R2 is ¨F. In some embodiments, R2 is ¨Cl. In some embodiments, R2 is ¨CN. In some embodiments, R2 is ¨0R3, wherein R3 is as described herein.

[0043] In some embodiments, each R3 is independently C1-C6 alkyl, wherein the alkyl is optionally substituted with one or more halogen. In some embodiments, R3 is Ci-C6 alkyl, wherein the alkyl is substituted with one halogen. In some embodiments, R3 is Ci-C6 alkyl, wherein the alkyl is substituted with two halogens. In some embodiments, R3 is selected from ¨CH3, ¨CHF2, and propyl. In some embodiments, R3 is ¨CH3. In some embodiments, R3 is ¨CHF2. In some embodiments, R3 is propyl.

[0044] In some embodiments, n is selected from 0, 1, 2, and 3. In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3.

[0045] In some embodiments, n is 2 and R2 is CH3 in one instance and F in the second instance.
In some embodiments, n is 2 and R2 is ¨OCH3 in one instance and Cl in the second instance. In some embodiments, n is 2 and R2 is ¨OCH3 in one instance and ¨OCH3 in the second instance. In some embodiments, n is 2 and R2 is ¨CH3 in one instance and F in the second instance. In some embodiments, n is 2 and R2 is F in one instance and F in the second instance.
In some embodiments, n is 2 and R2 is ¨CHF2 in one instance and F in the second instance. In some embodiments, n is 2 and R2 is ¨OCH(CH3)2 in one instance and F in the second instance. In some embodiments, n is 2 and R2 is ¨OCHF2 in one instance and F in the second instance. In some embodiments, n is 3 and R2 is ¨OCH3 in one instance, F in the second instance, and F in the third instance. In some embodiments, n is 3 and R2 is ¨OCH3 in one instance, ¨OCH3 in the second instance, and F in the third instance.

[0046] In some embodiments, R1 is H and n is 0.

[0047] In some embodiments, R1 is ¨OH and n is 0.

[0048] In some embodiments, R1 is ¨OH, n is 2, and R2 is ¨F in one instance and ¨0R3 in the second instance, wherein R3 is ¨CH3.

[0049] In some embodiments, R1 is ¨OH, n is 2, and R2 is ¨F in one instance and ¨0R3 in the second instance, wherein R3 is ¨CHF2.

[0050] In some embodiments, the compound of formula (I) is Compound 1:

F ii 01 OH
N¨

No.9..._ N

I (1) or a pharmaceutically acceptable salt thereof.

[0051] In some embodiments, the compound of formula (I) is (1R,3R)-3-[(7S)-2-[(R)-(5-fluoro-2-methoxyphenyl)(hydroxy)methyl]-6-(methoxycarbony1)-7-methyl-3H,6H,7H,8H,9H-imidazo[4,5-fiquinolin-3-yl]cyclohexane-l-carboxylic acid.

[0052] In some embodiments, the compound of formula (I) is (1R,3R)-3-((S)-2-((R)-(5-fluoro-2-methoxyphenyl)(hydroxy)methyl)-6-(methoxycarbonyl)-7-methyl-6,7,8,9-tetrahydro-imidazo[4,5-fiquinolin-3-y1)cyclohexane-1-carboxylic acid.

[0053] In some embodiments, the compound of formula (I) is Compound 2:
*
OH
N¨

Ni,.9..
N

I (2) or a pharmaceutically acceptable salt thereof.

[0054] In some embodiments, the compound of formula (I) is 3-((7S)-2-(hydroxy(phenyl)methyl)-6-(methoxycarbony1)-7-methy1-6,7,8,9-tetrahydro-3H-imidazo[4,5-fiquinolin-3-y1)cyclohexane-1-carboxylic acid.

[0055] In some embodiments, the compound of formula (I) is the first eluting isomer of 3-((7S)-2-(hydroxy(phenyl)methyl)-6-(methoxycarbony1)-7-methyl-6,7,8,9-tetrahydro-3H-imidazo[4,5-fiquinolin-3-y1)cyclohexane-1-carboxylic acid when eluted from a preparative HPLC using the conditions defined in Example 1.3.

[0056] In some embodiments, the compound of formula (I) is (1R,3R)-3-((S)-2-((R)-hydroxy(phenyl)methyl)-6-(methoxycarbony1)-7-methyl-6,7,8,9-tetrahydro-3H-imidazo[4,5-fiquinolin-3-y1)cyclohexane-1-carboxylic acid.

[0057] In some embodiments, the compound of formula (I) is Compound 3:
F 4.0 0>-F
OH
N-(3) or a pharmaceutically acceptable salt thereof.

[0058] In some embodiments, the compound of formula (I) is 3-((7S)-2-((2-(difluoromethoxy)-5-fluorophenyl)(hydroxy)methyl)-6-(methoxycarbonyl)-7-methyl-6,7,8,9-tetrahydro-imidazo[4,5-fiquinolin-3-y1)cyclohexane-1-carboxylic acid.

[0059] In some embodiments, the compound of formula (I) is the first eluting isomer of 3-((7S)-2-((2-(difluoromethoxy)-5-fluorophenyl)(hydroxy)methyl)-6-(methoxycarbonyl)-7-methyl-6,7,8,9-tetrahydro-3H-imidazo[4,5-fiquinolin-3-y1)cyclohexane-1-carboxylic acid when eluted from a preparative HPLC using the conditions defined in Example 1.5.

[0060] In some embodiments, the compound of formula (I) is (1R,3R)-3-((S)-2-((R)-(2-(difluoromethoxy)-5-fluorophenyl)(hydroxy)methyl)-6-(methoxycarbonyl)-7-methyl-6,7,8,9-tetrahydro-3H-imidazo[4,5-fiquinolin-3-y1)cyclohexane-1-carboxylic acid.

[0061] In some embodiments, the compound of formula (I) is Compound 4:

OH

(4) or a pharmaceutically acceptable salt thereof.

[0062] In some embodiments, the compound of formula (I) is 3-((S)-2-benzy1-6-(methoxyc arbony1)-7-methy1-6,7 ,8,9-tetrahydro-3H-imidazo [4,5-f] quinolin-3 -yl)c yclohex ane-1-carboxylic acid.

[0063] In some embodiments, the compound of formula (I) is (1R,3R)-3-((S)-2-benzy1-6-(methoxyc arbony1)-7-methy1-6,7 ,8,9-tetrahydro-3H-imidazo [4,5-f] quinolin-3 -yl)c yclohex ane-1-carboxylic acid.

[0064] The disclosure is also based in part on the recognition that compounds of formula (I) are Selective CBP Inhibitor Compounds, defined herein as CBP Inhibitors having a BRD4 IC50 value greater than that of their CBP IC50 value, preferably wherein its BRD4 IC50 value is greater than 1 i.tM (e.g., 1 micromolar to 10 micomolar, or greater), wherein the IC50 values are determined as in the procedures set forth in the assay described in Example 2. In some embodiments, compounds of formula (I) can be Selective CBP Inhibitor Compounds, wherein the BRD4 IC50 value is greater than 500 nM (e.g., 500 nanomolar to 10 micomolar, or greater), wherein the IC50 values are determined as in the procedures set forth in the assay described in Example 2.
The disclosure is also based in part on the recognition that Compound 1 is a Selective CBP
Inhibitor Compound, defined herein as a CBP Inhibitor having a BRD4 IC50 value greater than that of its CBP IC50 value, preferably wherein its BRD4 IC50 value greater than 1 i.tM (e.g., 1 micromolar to 10 micomolar, or greater), wherein the IC50 values are determined as in the procedures set forth in the assay described in Example 2.

[0065] The discovery includes the use of one or more compounds of formula (I), and pharmaceutically acceptable salts thereof, in pharmaceutical preparations for the treatment of patients diagnosed with a disease or disorder associated with the inhibition of CBP (e.g., certain forms of cancer). The compositions comprising one or more compounds of formula (I), and pharmaceutically acceptable salts thereof, can be obtained by certain processes also provided herein. In some embodiments, a Selective CBP Inhibitor Compound of formula (I) is used to treat breast cancer (e.g., TNBC) or prostate cancer. In some embodiments, a Selective CBP Inhibitor Compound of Formula (Ib) is used to treat an AR+ form of cancer, including AR+
breast cancer or prostate cancer. The use of a Selective CBP Inhibitor Compound of formula (I) is provided for treatment of a patient diagnosed with a AR+ form of cancer, such as AR+ breast cancer (e.g., AR+
TNBC) or AR+ prostate cancer (e.g., a AR-v7+ form of prostate cancer).

[0066] In some embodiments, the discovery includes the use of (1R,3R)-3-[(7S)-2-[(R)-(5-fluoro-2-methoxyphenyl)(hydroxy)methyll-6-(methoxycarbony1)7-methyl-3H,6H,7H,8H,9H-imidazo [4,5-f] quinolin-3-yl] cyclohexane- 1-carboxylic acid (Compound 1), and pharmaceutically acceptable salts thereof, in pharmaceutical preparations for the treatment of patients diagnosed with a disease or disorder associated with the inhibition of CBP (e.g., certain forms of cancer).
The compositions comprising Compound 1 and pharmaceutically acceptable salts thereof can be obtained by certain processes also provided herein.

[0067] In some embodiments, the discovery includes the use of Compound 2, and pharmaceutically acceptable salts thereof, in pharmaceutical preparations for the treatment of patients diagnosed with a disease or disorder associated with the inhibition of CBP (e.g., certain forms of cancer). The compositions comprising Compound 2 and pharmaceutically acceptable salts thereof can be obtained by certain processes also provided herein.

[0068] In some embodiments, the discovery includes the use of Compound 3, and pharmaceutically acceptable salts thereof, in pharmaceutical preparations for the treatment of patients diagnosed with a disease or disorder associated with the inhibition of CBP (e.g., certain forms of cancer). The compositions comprising Compound 3 and pharmaceutically acceptable salts thereof can be obtained by certain processes also provided herein.

[0069] In some embodiments, the discovery includes the use of Compound 4, and pharmaceutically acceptable salts thereof, in pharmaceutical preparations for the treatment of patients diagnosed with a disease or disorder associated with the inhibition of CBP (e.g., certain forms of cancer). The compositions comprising Compound 4 and pharmaceutically acceptable salts thereof can be obtained by certain processes also provided herein.
Methods of Synthesizing the Compounds

[0070] The compounds of the present disclosure may be made by a variety of methods, including standard chemistry. Suitable synthetic routes are depicted in the examples given below.

[0071] The compounds of the present disclosure, i.e., compounds of Formula (I), (II), or Group A
or a pharmaceutically acceptable salt thereof, may be prepared by methods known in the art of organic synthesis as set forth in part by the synthetic schemes depicted in the examples. In the schemes described below, it is well understood that protecting groups for sensitive or reactive groups are employed where necessary in accordance with general principles or chemistry.

Protecting groups are manipulated according to standard methods of organic synthesis (T. W.
Greene and P. G. M. Wuts, "Protective Groups in Organic Synthesis", Third edition, Wiley, New York 1999). These groups are removed at a convenient stage of the compound synthesis using methods that are readily apparent to those skilled in the art. The selection processes, as well as the reaction conditions and order of their execution, shall be consistent with the preparation of compounds of Formula (I), (II), or Group A.

[0072] Those skilled in the art will recognize stereocenters exist in the compounds of Formula (I), (II), or Group A. Accordingly, the present disclosure includes both possible stereoisomers (unless otherwise indicated and/or specified in the synthesis) and includes not only racemic compounds but the individual enantiomers and/or diastereomers as well. Unless otherwise indicated, when a compound is desired as a single enantiomer or diastereomer, it may be obtained by stereospecific synthesis or by resolution of the final product or any convenient intermediate. Resolution of the final product, an intermediate, or a starting material may be affected by any suitable method known in the art. See, for example, "Stereochemistry of Organic Compounds" by E. L.
Eliel, S. H. Wilen, and L. N. Mander (Wiley-lnterscience, 1994).
Methods of Using the Compounds

[0073] In some embodiments, compounds of formula (I) are tool compounds useful for studying the effects of CBP/p300 inhibition in vitro or in an in vivo model. In vitro, the tool compounds of formula (I) may be useful for studying the effects of CBP/p300 inhibition on purified proteins, cellular extracts, in intact cells and cell line models, and the like. In vivo, the tool compounds of formula (I) may be useful for studying the effects of CBP/p300 inhibition in cell line derived xenografts, in patient derived xenografts, in knock-in mouse model, in knock-out mouse models, and the like.

[0074] Preferably, the disclosure provides pharmaceutical preparations for the treatment of patients diagnosed with AR+ cancer. In particular, compounds provided herein can be formulated as an active pharmaceutical composition comprising one or more compounds of formula (I) (or a pharmaceutically acceptable salt and/or enantiomer thereof) useful for treatment of prostate cancer, including metastatic castrate resistant prostate cancer (CRPC), and/or AR+ breast cancers including locally advanced or metastatic AR+ breast cancer. For example, the inhibition of CBP/P300 can target AR transcriptional activity through H3K27Ac, reduction of AR target gene expression, or reduction of AR expression with, ultimately, a reduction in proliferation. In addition, CBP/P300 BRD inhibitors present the possibility of suppressing ER-driven signaling in hormone-receptor positive breast cancers. In some embodiments, the pharmaceutical composition comprises Compound 1. In some embodiments, the pharmaceutical composition comprises Compound 2. In some embodiments, the pharmaceutical composition comprises Compound 3. In some embodiments, the pharmaceutical composition comprises Compound 4.

[0075] Compounds and compositions described herein are inhibitors of CBP
having a lower inhibition concentration compared to its inhibition concentration with respect to BRD4.

[0076] Methods of treatment (e.g., by inhibiting CBP) can comprise administering to a subject in need thereof a therapeutically effective amount of (1) a compound of formula (I) or a pharmaceutically acceptable salt thereof; (2) a pharmaceutical composition comprising one or more compounds of formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0077] Methods of treatment (e.g., by inhibiting CBP) can comprise administering to a subject in need thereof a therapeutically effective amount of (1) Compound 1 or a pharmaceutically acceptable salt thereof; (2) a pharmaceutical composition comprising Compound 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0078] Methods of treatment (e.g., by inhibiting CBP) can comprise administering to a subject in need thereof a therapeutically effective amount of (1) Compound 2 or a pharmaceutically acceptable salt thereof; (2) a pharmaceutical composition comprising Compound 2 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0079] Methods of treatment (e.g., by inhibiting CBP) can comprise administering to a subject in need thereof a therapeutically effective amount of (1) Compound 3 or a pharmaceutically acceptable salt thereof; (2) a pharmaceutical composition comprising Compound 3 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0080] Methods of treatment (e.g., by inhibiting CBP) can comprise administering to a subject in need thereof a therapeutically effective amount of (1) Compound 4 or a pharmaceutically acceptable salt thereof; (2) a pharmaceutical composition comprising Compound 4 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0081] The pharmaceutical compositions may be orally administered in any orally acceptable dosage form. Accordingly, a patient and/or subject can be selected for treatment using a compound described herein by first evaluating the patient and/or subject to determine whether the subject is in need of inhibition of CBP, and if the subject is determined to be in need of inhibition of CBP, then administering to the subject a composition described herein.

[0082] The CBP Inhibitor compounds and compositions are useful, for example, in suppressing AR-driven transcriptional programs through inhibition of the CBP/P300 BRD, including AR-v7 variants. The growth inhibitory effect of Compounds 1, 2 and 4 and enzalutamide was determined across a panel of prostate cancer cell lines including androgen-dependent and independent models as well as AR-negative cell lines. Compound 1 induced a concentration-dependent reduction of H3K27Ac, a mark specific to CBP/P300, in an AR positive breast cancer cell line (Figure 3).
Compounds 1, 2, and 4 reduced the mRNA expression of TMPRSS2 and XBP1 in an AR
positive breast cancer cell line. Compound 1, 2 and 4 inhibit proliferation of breast cancer cell lines after days continuous exposure to the drug, and the cell lines with high expression of AR mRNA are more sensitive than those with low expression. Compound 1 treatment produced a tumor growth inhibition in an AR positive breast cancer cell line derived xenograft model (Figure 4). In some embodiments, an AR positive breast cancer cell line can be MDA-MB-453.
Treatment of prostate cancer cells with Compound 2 led to the reduction of both full length and variant forms of the AR
including AR-v7 (Figure 5). Compounds 1, 2, and 4 reduced AR target genes TMPRSS2 and KLK3, as well as MYC in a concentration-dependent manner in an AR-v7+ prostate cancer cell line. In some embodiments, an AR-v7+ prostate cancer cell line can be 22Rv1.
Compounds 1, 2, and 4 had a potent and concentration-dependent growth inhibitory effect in all AR+ cell lines, including AR-v7 harboring cell lines. Treatment with Compound 4 at 40 mg/kg/dose daily Monday - Thursday repeated weekly or 80 mg/kg/dose Monday and Thursday (twice weekly) repeated weekly resulted in a strong antitumor response in a patient-derived xenograft model resistant to enzalutamide (Figure 6).

[0083] Enzalutamide is an androgen receptor inhibitor indicated for the treatment of patients with castration resistant prostate cancer or metastatic castration-sensitive prostate cancer. Patients receiving enzalutamide can also receive a gonadotropin-releasing hormone (GnRH) analog concurrently or can have had bilateral orchiectomy. Enzalutamide is an androgen receptor inhibitor that acts on different steps in the androgen receptor signaling pathway.
Enzalutamide has been shown to competitively inhibit androgen binding to androgen receptors; and consequently, inhibits nuclear translocation of androgen receptors and their interaction with DNA. A
major metabolite, N-desmethyl enzalutamide, exhibited similar in vitro activity to enzalutamide.
Enzalutamide decreased proliferation and induced cell death of prostate cancer cells in vitro, and decreased tumor volume in a mouse prostate cancer xenograft model.

[0084] In some embodiments, compounds of formula (I), or pharmaceutically acceptable salts thereof, are indicated for the treatment of patients with castration resistant prostate cancer or metastatic castration-sensitive prostate cancer. Patients receiving a compound of formula (I), or a pharmaceutically acceptable salt thereof, can also receive gonadotropin-releasing hormone (GnRH) analog concurrently or can have had a bilateral orchiectomy.

[0085] Enzalutamide was active against androgen-dependent cell lines such as VCaP but was inactive in AR- negative cell lines as well as AR-v7+ cells. Conversely, Compound 1 had a potent and concentration-dependent growth inhibitory effect in all AR+ cell lines including an AR-v7+
cell line (IC50 =0.6 i.t1\4). Compound 1 was also inactive against AR- cell lines. This is consistent with the proposed mechanism of action of Compound 1.

[0086] Without being bound to theory, it is believed that CBP/P300 can interact with AR directly via the NR1D domain in CBP/P300. However, contrary to other nuclear receptors, this interaction is not believed to be dependent on ligand binding. Also, CBP/P300 can interact with both the LBD
of AR and its N-terminal domain. Both these factors are of relevance in castrate resistant prostate cancer and it is expected that CBP/P300 interacts with AR-v7 very similarly as with AR.
CBP/P300 also interact with AR indirectly via the co-factor TIP60/SRC-1 which itself interacts with AR. This approach can differ from direct receptor antagonists and can have the advantage of being unaffected by structural variations in the AR ligand binding domain (LBD). Such CBP/P300 BRD inhibitors can have activity in a number of cancers dependent upon nuclear hormone-receptor transcriptional programs, such as metastatic CRPC and locally advanced or metastatic AR+ breast cancers. The therapeutic benefits of AR inhibition have been demonstrated clinically in these cancers, but with limitations that highlight the need for alternative approaches with long term benefit and mechanisms of resistance that are non-overlapping with anti-androgen therapy. For example, the inhibition of CBP/P300 can target AR transcriptional activity through reduction of H3K27 acetylation, reduction of AR target gene expression, and/or reduction of AR expression, ultimately leading to a reduction in proliferation. CBP/P300 inhibitor-mediated inhibition of AR

activities is expected to be insensitive to AR LBD structural variations, and thus insensitive to LBD-related mechanisms of resistance to androgen antagonists. Finally, inhibitors can be useful for suppressing ER-driven signaling in hormone-receptor positive breast cancers.

[0087] AR aberrations in the LBD (splice variant v7, AR mutations) can render the receptor ligand-independent and insensitive to AR antagonists. Approximately twenty AR
mRNA splice variants have been identified, with a subset that are constitutively active.
Notably, all biologically active forms of the AR retain the NTD; drugs that target the NTD have the potential to impact all AR forms, including those that may drive resistance to AR-LBD-targeting therapies. Of the AR
variants, only AR-v7 and ARv567es have been detected at the protein level and AR-v7 has been the most studied. Notably, in cases of mCRPC where men were initially treated with an AR
antagonist, those with AR-v7-positive circulating tumor cells (CTC) showed poorer PSA response, and shorter PFS and OS, compared to those negative for AR-v7 CTCs.
Furthermore, in blood samples from mCRPC patients, the frequency of AR-v7 protein detection in CTC
nuclei increased from 3% of samples from patients following first line of therapy to 31% of samples for the third or more lines of therapy. Findings such as these point to the potential for using AR-v7 as a patient selection biomarker, and its likely utility for determining which men with mCRPC may benefit from AR antagonist treatment versus chemotherapy.

[0088] Various methods for measuring and defining AR positivity can be employed to select patients to receive a pharmaceutical composition comprising one or more compounds of formula (I). In the case of prostate cancer, AR expression is largely maintained in the target population of relapse patients. Furthermore, AR and AR-v7 protein or mRNA can be detected in CTC while AR
mutations and AR amplification can be detected from circulating tumor DNA
(ctDNA). Current IHC methods for measuring AR in breast cancer vary on multiple factors including the antibody used, the IHC methodology, and the cut-off criterion for positivity (Safarpour et al., Am. J. Cancer Res., 2014, 4:353-368). Most of the studies use the AR441 antibody clone from Dako (Agilent) and 1% or 10% positive nuclei as the threshold for positivity. Overall, the frequency of AR+ TNBC
has been reported to be 20% to 30%.

[0089] Prostate cancers resistant to androgen deprivation or androgen antagonists remain an unmet need. In some examples, the inhibition of CBP/P300 BRD, with its differentiated mechanism of antagonizing the AR-driven transcriptional program, is used as a treatment for these patients, with potential utility as an earlier line of therapy. Pharmaceutical compositions comprising one or more compounds of formula (I) can be used to treat certain forms of prostate cancer. Preferably, compositions comprising one or more compounds of formula (I) are useful for the inhibition of CBP/P300 BRD, with a differentiated mechanism of antagonizing the AR-driven transcriptional program (e.g., as an early line of therapy). Pharmaceutical compositions comprising one or more compounds of formula (I) can be used to treat mCRPC, or other cancers dependent on AR-driven transcription.

[0090] Some methods include administering a therapeutically effective amount of a pharmaceutical composition comprising one or more compounds of formula (I) to patients diagnosed with mCRPC with progressive castration-resistant disease who have failed or been intolerant to at least two prior systemic therapies, including at least one androgen antagonist-based therapy with evaluable disease (anti-androgen+LHRH analog, enzalutamide, or abiraterone) and either a rising PSA with or without detectable metastatic disease per standard definitions or neuroendocrine features. Compounds of formula (I) are useful for treatment of patients diagnosed with an AR-v-7 variant form of AR. In some methods, patients diagnosed with mCRPC with AR-v7 positive circulating tumor cells (CTC) can be treated with the pharmaceutical composition comprising one or more compounds of formula (I). In some methods, patients diagnosed with disease progression after treatment with enzalutamide can be treated with the pharmaceutical composition comprising one or more compounds of formula (I).

[0091] Some methods include administering a therapeutically effective amount of a pharmaceutical composition comprising Compound 1 to patients diagnosed with mCRPC with progressive castration-resistant disease who have failed or been intolerant to at least two prior systemic therapies, including at least one androgen antagonist-based therapy with evaluable disease (anti-androgen+LHRH analog, enzalutamide, or abiraterone) and either a rising PSA with or without detectable metastatic disease per standard definitions or neuroendocrine features.
Compound 1 is also useful for treatment of patients diagnosed with an AR-v-7 variant form of AR.
In some methods, patients diagnosed with mCRPC with AR-v7 positive circulating tumor cells (CTC) can be treated with the pharmaceutical composition comprising Compound 1. In some methods, patients diagnosed with disease progression after treatment with enzalutamide can be treated with the pharmaceutical composition comprising Compound 1.

92 PCT/US2020/022823 [0092] Some methods include administering a therapeutically effective amount of a pharmaceutical composition comprising (1R,3R)-3-((S )-2-((R)-(5-fluoro-2-methoxyphenyl)(hydroxy)methyl)-6-(methoxycarbonyl)-7-methyl-6,7,8,9-tetrahydro-imidazo[4,5-fiquinolin-3-y1)cyclohexane-1-carboxylic acid to patients diagnosed with mCRPC
with progressive castration-resistant disease who have failed or been intolerant to at least two prior systemic therapies, including at least one androgen antagonist-based therapy with evaluable disease (anti-androgen+LHRH analog, enzalutamide, or abiraterone) and either a rising PSA with or without detectable metastatic disease per standard definitions or neuroendocrine features.
(1R,3R)-3-((S)-2-((R)-(5-fluoro-2-methoxyphenyl)(hydroxy)methyl)-6-(methoxycarbonyl)-7-methyl-6,7,8,9-tetrahydro-3H-imidazo [4,5-f] quinolin-3 -yl)c yclohexane- 1-carboxylic acid is also useful for treatment of patients diagnosed with an AR-v-7 variant form of AR.
In some methods, patients diagnosed with mCRPC with AR-v7 positive circulating tumor cells (CTC) can be treated with the pharmaceutical composition comprising (1R,3R)-3-((S)-2-((R)-(5-fluoro-methoxyphenyl)(hydroxy)methyl)-6-(methoxycarbonyl)-7-methyl-6,7,8,9-tetrahydro-imidazo [4,5-f] quinolin-3-yl)c yclohex ane-1 -c arbo xylic acid. In some methods, patients diagnosed with disease progression after treatment with enzalutamide can be treated with the pharmaceutical composition comprising (1R,3R)-3-((S)-2-((R)-(5-fluoro-2-methoxyphenyl)(hydroxy)methyl)-6-(methoxycarbonyl)-7-methyl-6,7,8,9-tetrahydro-3H-imidazo [4,5-f] quinolin-3 -yl)c yclohex ane-1-carboxylic acid.

[0093] Some methods include administering a therapeutically effective amount of a pharmaceutical composition comprising Compound 2 to patients diagnosed with mCRPC with progressive castration-resistant disease who have failed or been intolerant to at least two prior systemic therapies, including at least one androgen antagonist-based therapy with evaluable disease (anti-androgen+LHRH analog, enzalutamide, or abiraterone) and either a rising PSA with or without detectable metastatic disease per standard definitions or neuroendocrine features.
Compound 2 is also useful for treatment of patients diagnosed with an AR-v-7 variant form of AR.
In some methods, patients diagnosed with mCRPC with AR-v7 positive circulating tumor cells (CTC) can be treated with the pharmaceutical composition comprising Compound 2. In some methods, patients diagnosed with disease progression after treatment with enzalutamide can be treated with the pharmaceutical composition comprising Compound 2.

[0094] Some methods include administering a therapeutically effective amount of a pharmaceutical composition comprising Compound 3 to patients diagnosed with mCRPC with progressive castration-resistant disease who have failed or been intolerant to at least two prior systemic therapies, including at least one androgen antagonist-based therapy with evaluable disease (anti-androgen+LHRH analog, enzalutamide, or abiraterone) and either a rising PSA with or without detectable metastatic disease per standard definitions or neuroendocrine features.
Compound 3 is also useful for treatment of patients diagnosed with an AR-v-7 variant form of AR.
In some methods, patients diagnosed with mCRPC with AR-v7 positive circulating tumor cells (CTC) can be treated with the pharmaceutical composition comprising Compound 3. In some methods, patients diagnosed with disease progression after treatment with enzalutamide can be treated with the pharmaceutical composition comprising Compound 3.

[0095] Some methods include administering a therapeutically effective amount of a pharmaceutical composition comprising Compound 4 to patients diagnosed with mCRPC with progressive castration-resistant disease who have failed or been intolerant to at least two prior systemic therapies, including at least one androgen antagonist-based therapy with evaluable disease (anti-androgen+LHRH analog, enzalutamide, or abiraterone) and either a rising PSA with or without detectable metastatic disease per standard definitions or neuroendocrine features.
Compound 4 is also useful for treatment of patients diagnosed with an AR-v-7 variant form of AR.
In some methods, patients diagnosed with mCRPC with AR-v7 positive circulating tumor cells (CTC) can be treated with the pharmaceutical composition comprising Compound 4. In some methods, patients diagnosed with disease progression after treatment with enzalutamide can be treated with the pharmaceutical composition comprising Compound 4.

[0096] Some methods include administering a therapeutically effective amount of a pharmaceutical composition comprising (1R,3R)-3-((S )-2-benzy1-6-(methoxycarbony1)-7-methy1-6,7 ,8,9-tetrahydro-3H-imidazo [4,5-f] quinolin-3 -yl)c yclohexane- 1-carboxylic acid to patients diagnosed with mCRPC with progressive castration-resistant disease who have failed or been intolerant to at least two prior systemic therapies, including at least one androgen antagonist-based therapy with evaluable disease (anti-androgen+LHRH analog, enzalutamide, or abiraterone) and either a rising PSA with or without detectable metastatic disease per standard definitions or neuroendocrine features. (1R,3R)-3-((S)-2-benzy1-6-(methoxycarbony1)-7-methyl-6,7,8,9-tetrahydro-3H-imidazo[4,5-fiquinolin-3-y1)cyclohexane-1-carboxylic acid is also useful for treatment of patients diagnosed with an AR-v-7 variant form of AR. In some methods, patients diagnosed with mCRPC with AR-v7 positive circulating tumor cells (CTC) can be treated with the pharmaceutical composition comprising (1R,3R)-3-((S )-2-benzy1-6-(methoxycarbony1)-7-methy1-6,7 ,8,9-tetrahydro-3H-imidazo [4,5-f] quinolin-3 -yl)c yclohexane- 1-carboxylic acid. In some methods, patients diagnosed with disease progression after treatment with enzalutamide can be treated with the pharmaceutical composition comprising (1R,3R)-3-((S)-2-benzy1-6-(methoxyc arbony1)-7-methy1-6,7 ,8,9-tetrahydro-3 H-imidazo [4,5-f] quinolin-3 -yl)c yclohex ane-1-carboxylic acid.

[0097] Pharmaceutical compositions comprising one or more compounds of formula (I) are useful for the treatment of certain forms of AR+ breast cancer. For example, the suppression of AR
and/or ER transcriptional activity with a compound of formula (I) (or a pharmaceutical composition comprising one or more compounds of formula (I)) can be useful in treating antitumor effects in AR+ breast cancers, including TNBC and metastatic ER+ tumors, via its inhibition of CBP/P300 BRD. In some embodiments, the suppression of AR and/or ER
transcriptional activity with a compound of formula (I) (or a pharmaceutical composition comprising one or more compounds of formula (I)) can be useful in treating antitumor effects in AR+
breast cancers, including TNBC and ER+ tumors, via its inhibition of CBP/P300 BRD. In some embodiments, the suppression of AR and/or ER transcriptional activity with Compound 1 (or a pharmaceutical composition comprising Compound 1) can be useful in treating antitumor effects in AR+ breast cancers, including TNBC and metastatic ER+ tumors, via its inhibition of CBP/P300 BRD. In some embodiments, the suppression of AR and/or ER transcriptional activity with Compound 1 (or a pharmaceutical composition comprising Compound 1) can be useful in treating antitumor effects in AR+ breast cancers, including TNBC and ER+ tumors, via its inhibition of CBP/P300 BRD. In some embodiments, the suppression of AR and/or ER transcriptional activity with Compound 2 (or a pharmaceutical composition comprising Compound 2) can be useful in treating antitumor effects in AR+ breast cancers, including TNBC and ER+ tumors, via its inhibition of CBP/P300 BRD. In some embodiments, the suppression of AR and/or ER
transcriptional activity with Compound 3 (or a pharmaceutical composition comprising Compound 3) can be useful in treating antitumor effects in AR+ breast cancers, including TNBC and ER+
tumors, via its inhibition of CBP/P300 BRD. In some embodiments, the suppression of AR and/or ER
transcriptional activity with Compound 4 (or a pharmaceutical composition comprising Compound 4) can be useful in treating antitumor effects in AR+ breast cancers, including TNBC and ER+
tumors, via its inhibition of CBP/P300 BRD.

[0098] Some methods include administering a therapeutically effective amount of a pharmaceutical composition comprising one or more compounds of formula (I) to patients diagnosed with invasive breast carcinoma with triple negative status (per College of American Pathologists [CAP] guidelines) and detectable AR expression in >1% of tumor cells, with progressive disease who has failed at least two prior systemic therapies with evaluable disease or invasive breast carcinoma AR positive >1% and ER, PR, or HER positive (per CAP
guidelines) with progressive disease who has failed at least three prior systemic therapies. Some methods include administering a therapeutically effective amount of a pharmaceutical composition comprising one or more compounds of formula (I) to patients diagnosed with Her2- breast cancer.
Some methods include administering a therapeutically effective amount of a pharmaceutical composition comprising one or more compounds of formula (I) to patients diagnosed with ER-, PR-, or ER-/PR- breast cancer.

[0099] Some methods include administering a therapeutically effective amount of a pharmaceutical composition comprising Compound 1 to patients diagnosed with invasive breast carcinoma with triple negative status (per College of American Pathologists [CAP] guidelines) and detectable AR expression in >1% of tumor cells, with progressive disease who has failed at least two prior systemic therapies with evaluable disease or invasive breast carcinoma AR positive >1%
and ER, PR, or HER positive (per CAP guidelines) with progressive disease who has failed at least three prior systemic therapies. Some methods include administering a therapeutically effective amount of a pharmaceutical composition comprising Compound 1 to patients diagnosed with Her2- breast cancer. Some methods include administering a therapeutically effective amount of a pharmaceutical composition comprising Compound 1 to patients diagnosed with ER-, PR-, or ER-/PR- breast cancer.

[0100] Some methods include administering a therapeutically effective amount of a pharmaceutical composition comprising Compound 2 to patients diagnosed with invasive breast carcinoma with triple negative status (per College of American Pathologists [CAP] guidelines) and detectable AR expression in >1% of tumor cells, with progressive disease who has failed at least two prior systemic therapies with evaluable disease or invasive breast carcinoma AR positive >1%
and ER, PR, or HER positive (per CAP guidelines) with progressive disease who has failed at least three prior systemic therapies. Some methods include administering a therapeutically effective amount of a pharmaceutical composition comprising Compound 2 to patients diagnosed with Her2- breast cancer. Some methods include administering a therapeutically effective amount of a pharmaceutical composition comprising Compound 2 to patients diagnosed with ER-, PR-, or ER-/PR- breast cancer.

[0101] Some methods include administering a therapeutically effective amount of a pharmaceutical composition comprising Compound 3 to patients diagnosed with invasive breast carcinoma with triple negative status (per College of American Pathologists [CAP] guidelines) and detectable AR expression in >1% of tumor cells, with progressive disease who has failed at least two prior systemic therapies with evaluable disease or invasive breast carcinoma AR positive >1%
and ER, PR, or HER positive (per CAP guidelines) with progressive disease who has failed at least three prior systemic therapies. Some methods include administering a therapeutically effective amount of a pharmaceutical composition comprising Compound 3 to patients diagnosed with Her2- breast cancer. Some methods include administering a therapeutically effective amount of a pharmaceutical composition comprising Compound 3 to patients diagnosed with ER-, PR-, or ER-/PR- breast cancer.

[0102] Some methods include administering a therapeutically effective amount of a pharmaceutical composition comprising Compound 4 to patients diagnosed with invasive breast carcinoma with triple negative status (per College of American Pathologists [CAP] guidelines) and detectable AR expression in >1% of tumor cells, with progressive disease who has failed at least two prior systemic therapies with evaluable disease or invasive breast carcinoma AR positive >1%
and ER, PR, or HER positive (per CAP guidelines) with progressive disease who has failed at least three prior systemic therapies. Some methods include administering a therapeutically effective amount of a pharmaceutical composition comprising Compound 4 to patients diagnosed with Her2- breast cancer. Some methods include administering a therapeutically effective amount of a pharmaceutical composition comprising Compound 4 to patients diagnosed with ER-, PR-, or ER-/PR- breast cancer.

[0103] The present disclosure enables one of skill in the relevant art to make and use the inventions provided herein in accordance with multiple and varied embodiments. Various alterations, modifications, and improvements of the present disclosure that readily occur to those skilled in the art, including certain alterations, modifications, substitutions, and improvements are also part of this disclosure. Accordingly, the foregoing description are by way of example to illustrate the discoveries provided herein. The present disclosure provides compounds which are Selective CBP
Inhibitors.
EXAMPLES
Definitions used in the following Schemes and elsewhere herein are:
ACN acetonitrile Ac20 acetic anhydride (+)BINAP ( )-2,2'-Bis(diphenylphosphino)-1,1'-binaphthalen Boc tert-butoxycarbonyl n-BuOH butanol cm centimeter DCE 1,2-dichloroethane DCM dichloromethane or methylene chloride DEA diethylamine DMC 2-Chloro-4,5-dihydro- 1,3 -dimethy1-1H-imidazolium chloride DMP Dess-Martin periodinane DMTMM 4-(4,6-Dimethoxy-1,3,5-triazin-2-y1)-4-methylmorpholinium chloride DIEA N,N-diisopropylethylamine DMAP 4-(dimethylamino)pyridine DMF N,N-dimethylformamide DMS 0 dimethylsulfoxide DPPA diphenylphosphoryl azide dppf bis(diphenylphosphino)ferrocene ES electro spray ionization Et3N triethylamine Et0Ac ethyl acetate Et0H ethanol FA formic acid FCC flash column chromatography h hours HATU 2-(3H- [ 1,2,3 ] triazolo [4,5-b]pyridin-3 - y1)- 1, 1,3,3 -tetramethylisouronium hexafluorophosphate HC1 hydrogen chloride HOAc acetic acid HPLC high performance liquid chromatography (i-Pr)2NEt N,N-diisopropylethylamine L liter LC/MS liquid chromatography/mass spectrometry LDA lithium diisopropylamine K2CO3 potassium carbonate Me0H methanol mL milliliter mmol millimole mg milligram MHz megahertz MS mass spectrometry m/z mass/charge ratio NB S N-bromosuccinimide nm nanometer NMM 4-methylmorpholine NMR nuclear magnetic resonance Pd2(dba)3 tris(dibenzylideneacetone)dipalladium Ph3P triphenylphosphine PhCHO benzaldehyde PhMe toluene PPm parts per million rt room temperature RT rentention time SFC supercritical fluid chromatography STAB sodium triacetoxyborohydride p-TSA para-toluenesulfonic anhydride p-Ts0H para-toluenesulfonic acid TFA trifluoroacetic acid TFAA trifluoroacetic anhydride THF tetrahydrofuran UV ultraviolet XPhos 2-dic yclohexylpho sphino-2 1,4 ',6 '-triis oprop ylbiphenyl Materials

[0104] Unless otherwise noted, all materials were obtained from commercial suppliers and were used without further purification. Anhydrous solvents were obtained from Sigma-Aldrich (Milwaukee, WI) and used directly. All reactions involving air- or moisture¨sensitive reagents were performed under a nitrogen atmosphere and all reactions utilizing microwave irraditation were run on a Biotage Initiator EXP EU instrument.

[0105] Unless otherwise noted, mass-triggered HPLC purification and/or purity and low resolution mass spectral data were measured using either: (1) Waters Acquity ultra performance liquid chromatography (UPLC) system (Waters Acquity UPLC with Sample Organizer and Waters Micromass ZQ Mass Spectrometer) with UV detection at 220 nm and a low resonance electrospray positive ion mode (ESI) (Column: Acquity UPLC BEH C18 1.7i.tm 2.1 X 50 mm;
gradient: 5-100% Solvent B (95/5/0.09%: Acetonitrile/Water/Formic Acid) in Solvent A
(95/5/0.1%: 10mM Ammonium Formate/Acetonitrile/Formic Acid) for 2.2 min then 100-5%
Solvent B in Solvent A for 0.01 min then hold at 5% Solvent B in Solvent A for 0.29 min) or (2) Waters HT2790 Alliance high performance liquid chromatography (HPLC) system (Waters 996 PDA and Waters ZQ Single Quad Mass Spectrometer) with UV detection at 220 nm and 254 nm and a low resonance electrospray ionization (positive/negative) mode (ESI) (Column: XBridge Phenyl or C18, 5 iim 4.6x50 mm; gradient: 5-95% Solvent B (95% methanol/5%
water with 0.1%
Formic Acid) in Solvent A (95% water/5% methanol with 0.1% Formic Acid) for 2.5 min then hold at 95% Solvent B in Solvent A for 1 min (purity and low resolution MS
only).

General Methods of Compound Preparation

[0106] Described herein are methods of synthesizing the compounds of the present disclosure.
Compounds of the present disclosure can be synthesized according to the synthetic schemes provided below. Preparation of the starting material for Schemes 1 and 2 ("Intermediate 1") is described below. Preparation of the starting material for Schemes 3 and 4 can be found in Example 1, Part A of U.S. Patent No. 4,404,207.

[0107] Unless otherwise specified, the substituents R4 and R5 in the following reaction schemes are defined as follows.
( R2) ( R2),, (1,3 (1 _ R4 = OH or R5 = 1..._<:_!--OH

[0108] Scheme 1 provides methods useful for synthesizing compounds of Formula I.
Scheme]

Br R4-00CI is Br NH3 NH2 ,... _______________________________________________ ,...
N 40 ____________ N Cul, L-proline N

HOAc, H2SO4 Nr----( N.----N-R5 NaH, R5-X NH
--c _________________________________________________ N N

[0109] Scheme 2 provides methods useful for synthesizing compounds of Formula I.

Scheme 2 NH2 NH2 N=---( 0 Br NH2-R5 NH-R5 124-CHO N-R5 N Pd-based Catalyst N N

[NM Alternatively, Scheme 3 provides methods useful for synthesizing certain compounds of Formula I.
Scheme 3 F NH2-R5 NH-RS Fe, NH4CI NH-R5 /
N N N
cat . *
H2N-Ru-OTf R4 R4 , R4 N--.--( N----r( pho=NTs (s) (s) N----X
N-R5 CICO2CH3 N-Rs Ph /NR

s 4 __________________________________________ 4 _________ /
TJ
N N . FA, Et3N, Me0H N
H

[0111] Alternatively, Scheme 4 provides methods useful for synthesizing certain compounds of Formula I.
Scheme 4 1. H2, 12 NO2 (S)-(-)-Me0Biphep NO2 NO2 / F [Ir(COD)C1]2 F CICO2CH3 F
_______________________ v.- ________________ )...-N 2. D-CSA N N
H

,i R5-NH2 N--=-- NH2 NO2 N-R5 R4-CHO s NH-R5 H2 NH-R5 N Pd/C
N' N
0 0 0 0 0 o Example 1: Syntheses of Compounds of the Disclosure [0112] The compounds listed in Figure 1 were prepared using standard chemical manipulations and procedures similar to those described herein. In Figure 1, "Eluted Isomer"
refers to the order in which the compound eluted by preparative HPLC.
Example 1.1: Preparation of Intermediate 1: methyl (S)-5-amino-6-bromo-2-methyl-3,4-dihydroquinoline-1(2H)-carboxylate [0113] Figure 2(A) provides a synthetic scheme for the preparation of Intermediate 1, as described below.
Step 1. 8-chloro-5-methoxy-2-methylquinoline hydrochloride [0114] Into a 5L 4-necked round-bottom flask purged and maintained with an inert atmosphere of nitrogen, 2-chloro-5-methoxyaniline (250 g, 1.59 mol) was dissolved in 1-butanol (1200 mL).
Then hydrochloric acid (aq, 36.5%, 526.5 mL) and chloranil (456.5 g, 1.86 mol) were added. The resulting mixture was stirred for 1 h at 100 C under nitrogen atmosphere.
Then a solution of (E)-but-2-enal (169 mL, 2.06 mol) in 1-butanol (300 mL) was added dropwise. The resulting solution was stirred for 1 h at 100 C under nitrogen atmosphere. The oil bath was cooled to 70 C and tetrahydrofuran (1500mL) was added. Then the resulting mixture was stirred for 1 h at 70 C. The reaction mixture was cooled to 0 C and the solids were filtered.
The solids were washed with tetrahydrofuran (3L) at 0 C then dried in an oven to afford 8-chloro-5-methoxy-2-methylquinoline hydrochloride (83.0 g, 74%) as a yellow solid. MS (ES, m/z):
208 [M+H]t Step 2. 5-methoxy-2-methylquinoline [0115] Into a 1000-mL 3-necked round-bottom flask, 8-chloro-5-methoxy-2-methylquinoline hydrochloride (50 g, 204.82 mmol) was dissolved in methanol (300 mL). Then sodium hydroxide (3M, 205 mL) and 10% palladium on carbon (25 g) were added. Hydrogen (g) was charged into the reaction mixture. The reaction mixture was stirred under a hydrogen atmosphere for 3 h at room temperature. The reaction was vented to nitrogen and the solids were filtered out over celite.
The filtered solution was concentrated under vacuum. The residue was subjected to purification by FCC eluting with ethyl acetate/petroleum ether (1:5). This afforded the title compound (28.5 g, 80%) as a yellow oil. MS: (ES, m/z): 174 [M+H]t Step 3. (2S)-5-methoxy-2-methyl-1,2,3,4-tetrahydroquinoline [0116] Into a 30-mL pressure tank reactor (50 atm), 5-methoxy-2-methylquinoline (4.0 g, 23.09 mmol) was dissolved in methanol (10 mL). Then Ru(0Tf)(0-hexamethylbenzene)((S,S)-TsDPEN) 4N-R1S,25)-2-(amino-0)-1,2-diphenylethyl] -4-methylbenzene sulfonamidato-KATI [(1,2,3,4,5,6- q)-1,2,3,4,5,6-hexamethylbenzene] (1,1,1-trifluoromethane sulfonato-K0)-ruthenium, prepared according to the procedure in J. Am. Chem. Soc. 2011, 133, 9878-9891) (150 mg, 0.23 mmol) was added. To the above hydrogen was introduced in. The resulting solution was stirred for 6 h at room temperature. The resulting mixture was concentrated under vacuum. The residue was subjected to purification by FCC eluting with ethyl acetate/petroleum ether (1:4). This afforded the title compound (3.0 g, 73%) as a yellow oil. MS: (ES, m/z): 178 [M+H]t Step 4. methyl (S)-5-methoxy-2-methyl-3,4-dihydroquinoline-1(2H)-carboxylate [0117] Into a 250-mL round-bottom flask, (2S)-5-methoxy-2-methy1-1,2,3,4-tetrahydroquinoline (18 g, 99.52 mmol) was dissolved in dichloromethane (100 mL). Then pyridine (23.6 g, 298.36 mmol) was added, followed by methyl carbonochloridate (9.4 g, 99.47 mmol). The resulting solution was stirred for 1 h at room temperature. The resulting solution was diluted with 100 mL
of dichloromethane and washed with 3x200 mL of water. The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The residue was subjected to purification by FCC eluting with ethyl acetate/petroleum ether (1:3). This afforded the title compound (21 g, 89%) as a yellow oil. MS: (ES, m/z): 236 [M+H]t Step 5. methyl (S)-5-hydroxy-2-methyl-3,4-dihydroquinoline-1(2H)-carboxylate [0118] Into a 500-mL 3-necked round-bottom flask, methyl (2S)-5-methoxy-2-methy1-1,2,3,4-tetrahydroquinoline-l-carboxylate (21 g, 89.36 mmol) was dissolved in dichloromethane (150 mL). Then boron tribromide (150 mL, 0.15 mol, 1 M in CH2C12) was added. The resulting solution was stirred for 1 h at room temperature. The reaction was then quenched by the addition of 300 mL of water. The resulting mixture was extracted with 3x300 mL of dichloromethane. The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The residue was subjected to purification by FCC eluting with ethyl acetate/petroleum ether (1:2). This afforded the title compound (13.5 g, 68%) as a yellow solid.
MS: (ES, m/z): 222 [M+H] .
Step 6. methyl (S)-2-methyl-5-(((trifluoromethyl)sulfonyl)oxy)-3,4-dihydroquinoline-1(2H)-carboxylate [0119] Into a 250-mL round-bottom flask, methyl (2S)-5-hydroxy-2-methy1-1,2,3,4-tetrahydroquinoline-l-carboxylate (5 g, 18.08 mmol) was dissolved in dichloromethane (50 mL).
Then pyridine (14.3 g, 180.78 mmol) and trifluoromethanesulfonic anhydride (10.2 g, 36.15 mmol) were added. The resulting solution was stirred for 1 h at room temperature. The resulting mixture was washed with 3x100 mL of water. The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The residue was subjected to purification by FCC eluting with ethyl acetate/petroleum ether (1:3). This afforded the title compound (5.5 g, 86%) as a yellow oil. MS: (ES, m/z): 354 [M+H]t Step 7. methyl (S)-5-((diphenylmethylene)amino)-2-methyl-3,4-dihydroquinoline-1(2H)-carboxylate [0120] Into a 500-mL round-bottom flask purged and maintained with an inert atmosphere of nitrogen, methyl (2S)-2-methy1-5-[(trifluoromethane)sulfonyloxy]-1,2,3,4-tetrahydroquinoline -1-carboxylate (23.5 g, 65.18 mmol) was dissolved in toluene (100 mL). Then diphenylmethanimine (17.9 g, 97.78 mmol), tris(dibenzylideneacetone)dipalladium-chloroform adduct (1.19 g, 1.30 mmol), (+/-)-2,2'-Bis(diphenylphosphino)-1,1'-binaphthyl (2.43 g, 3.90 mmol) and cesium carbonate (42.4 g, 130.13 mmol) were added. The resulting solution was stirred overnight at 100 C under nitrogen atmosphere. The reaction mixture was cooled and the solids were filtered out. The residue was subjected to purification by FCC eluting with ethyl acetate/petroleum ether (1:3). This afforded the title compound (33 g, 80%) as a yellow oil. MS:
(ES, m/z): 385 [M+H]t Step 8. methyl (S)-5-amino-2-methyl-3,4-dihydroquinoline-1(2H)-carboxylate [0121] Into a 500-mL round-bottom flask, methyl (25)-5-[(diphenylmethylidene)amino]-2-methy1-1,2,3,4-tetrahydroquinoline-l-carboxylate (33 g, 85.93 mmol) was dissolved in methanol (200 mL). Then sodium acetate (17 g, 207.23 mmol) and hydroxylamine hydrochloride (12.3 g, 177.00 mmol) were added. The resulting solution was stirred for 2 h at room temperature. The solids were filtered out. The resulting mixture was concentrated under vacuum.
The residue was subjected to purification by FCC eluting with ethyl acetate/petroleum ether (1:2). This afforded the title compound (12.5 g, 66%) as a yellow solid. MS: (ES, m/z): 221 [M+H]t Step 9. methyl (S)-5-amino-6-bromo-2-methyl-3,4-dihydroquinoline-1(2H)-carboxylate (Intermediate 1) [0122] Into a 100-mL 3-necked round-bottom flask, methyl (25)-5-amino-2-methy1-1,2,3,4-tetrahydroquinoline-1-carboxylate (1 g, 4.09 mmol) was dissolved in acetonitrile (20 mL). Then N-bromosuccinimide (730 mg, 4.10 mmol) was added. The resulting solution was stirred for 30 min at room temperature. The resulting mixture was concentrated under vacuum.
The residue was subjected to purification by FCC eluting with ethyl acetate/petroleum ether (1:1). This afforded the title compound (1.1 g, 90%) as a yellow solid. MS: (ES, m/z): 299, 301 [M+H]t 1H-NMR: (400 MHz, CD30D, ppm): 7.19(d, J= 8.8 Hz, 1H), 6.84(d, J= 8.8 Hz, 1H), 4.73-4.69(m, 1H), 3.74(s, 3H), 2.64-2.57(m, 1H), 2.55-2.44(m, 1H), 2.12-2.05(m, 1H), 1.82-1.79(m, 1H), 1.17(d, J=6.9 Hz, 3H).
Example 1.2: Synthesis of (1R,3R)-3-[(7S)-2-[(R)-(5-fluoro-2-methoxyphenyl)(hydroxy)methy1]-6(methoxycarbony1)-7-methyl-3H,6H,7H,8H,9H-imidazo [4,54] quinolin-3 -yl] cyclohexane-1 carboxylic acid (1); (1R,3R)-3 -[(75 )- 2- [(S)- (5 -fluoro-2-methoxyphenyl)(hydroxy)methy1]-6(methoxycarbony1)-7-methyl-3H,6H,7H,8H,9H-imidazo[4,5-flquinolin-3-yl]cyclohexane-lcarboxylic acid (1') [0123] Figure 2(B) provides a synthetic scheme for the preparation of Compound 1 and Compound 1', as described below.
Synthesis of intermediate 2-(5-fluoro-2-methoxypheny1)-2-hydroxyacetic acid Step 1. 2-(5-fluoro-2-methoxypheny1)-2-ktrimethylsily1)oxylacetonitrile [0124] A solution of ZnI2 (1.6 mg, 0.01 mmol), 5-fluoro-2-methoxybenzaldehyde (1.54 g, 9.99 mmol) in trimethylsilanecarbonitrile (1.5 mL, 11.25 mmol) was stirred for 1 h at room temperature.
The resulting mixture was concentrated under vacuum. The resulting crude product was purified by silica gel chromatography (eluting with 1:1 ethyl acetate/petroleum ether) to afford 2-(5-fluoro-2-methoxypheny1)-2-[(trimethylsily1)oxy]acetonitrile as a white solid (2.0 g, 79%).
Step 2. 2-(5-fluoro-2-methoxypheny1)-2-hydroxyacetic acid [0125] A solution of 2-(5-fluoro-2-methoxypheny1)-2-[(trimethylsily1)oxy]acetonitrile (1.50 g, 5.92 mmol) in hydrochloric acid (10 mL, 12M was stirred for 1 h at 25 C, and then stirred for 2 h at 70 C. The reaction mixture was cooled and concentrated under vacuum. The crude product was purified by reverse phase chromatography (Column: C18; Mobile phase, A:
water (containing 0.05% TFA) and B: ACN (5% to 20% over 30 min); Detector, UV 254 nm) to afford 2-(5-fluoro-2-methoxypheny1)-2-hydroxyacetic acid as a white solid (1.10 g, 93%).
Step 3. 6-fluoro-2-methyl-5-nitroquinoline [0126] A solution of trifluoromethanesulfonic acid (82.0 mL, 0.923 mol) in HNO3 (19.6 mL, 0.437 mol) was stirred for 20 min at 0 C. This was followed by the addition of 6-fluoro-2-methylquinoline (50.0 g, 0.310 mol) in dichloromethane (300 mL) at 0 C. The resulting mixture was stirred for 15 h at room temperature (25 C). The reaction mixture was diluted with water (300 mL). The pH value of the solution was adjusted to 8 with sodium bicarbonate (saturated aqueous solution). The resulting solution was extracted with dichloromethane (3 x 300 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum.
The residue was purified by silica gel chromatography (eluting with 1:4 ethyl acetate/petroleum ether) to afford 6-fluoro-2-methyl-5-nitroquinoline as a light yellow solid (60.0 g, 94%). LCMS
(ES, m/z): 207 [M+H]t Step 4. (28)-6-fluoro-2-methyl-5-nitro-1,2,3,4-tetrahydroquinoline [0127] A solution of (S)-(-)-Me0-BIPHEP (1.03 g, 1.77 mmol), chloro(1,5-cyclooctadiene)iridium(I) dimer (538 mg, 0.80 mmol) in toluene (100 mL) was stirred for 30 min at room temperature (25 C) under an atmosphere of nitrogen. This was followed by the addition of 12 (410 mg, 1.62 mmol) and 6-fluoro-2-methyl-5-nitroquinoline (33.0 g, 0.160 mol) in toluene (100 mL). The resulting mixture was stirred for 20 h at room temperature (25 C) under hydrogen (50 atm). The resulting mixture was concentrated under vacuum and purified by silica gel chromatography (eluting with 1:1 ethyl acetate/petroleum ether) to afford the crude product (35.0 g). The crude product was dissolved in ethyl acetate (230 mL), followed by the addition of D-Camphorsulfonic acid (36.9 g, 0.158 mol). The resulting solution was stirred for 1 h at 60 C and then cooled to room temperature. The solids were collected by filtration, and rinsed with ethyl acetate (120 mL). The solids were dissolved in water (50 mL). The pH value of the solution was adjusted to 8 with sodium bicarbonate (saturated aqueous solution). The resulting solution was extracted with ethyl acetate (3 x 120 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to afford (25)-6-fluoro-2-methy1-5-nitro-1,2,3,4-tetrahydroquinoline as a red solid (25.5 g, 76%). LCMS (ES, m/z): 211 [M+H]
Step 5. methyl (28)-6-fluoro-2-methyl-5-nitro-1,2,3,4-tetrahydroquinoline-1-carboxylate [0128] A solution of (2S )-6-fluoro-2-methyl-5-nitro-1,2,3,4-tetrahydroquinoline (25.3 g, 0.120 mol), pyridine (39.0 mL, 0.484 mol), methyl carbonochloridate (18.7 mL, 0.242 mol) in dichloromethane (150 mL) was stirred for 3 h at room temperature (25 C). The reaction was washed with 1M hydrochloric acid (2 x 70 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to afford methyl (25)-6-fluoro-2-methy1-5-nitro-1,2,3,4-tetrahydroquinoline-1-carboxylate as a yellow solid (29.8 g, 92%).
LCMS (ES, m/z): 269 [M+H]t Step 6. methyl (2S)-6-[[(1R,3R)-3-(methoxycarbonyl)cyclohexyl]amino]-2-methyl-nitro1,2,3,4-tetrahydroquinoline-1-carboxylate [0129] A solution of methyl (2S )-6-fluoro-2-methy1-5-nitro-1,2,3,4-tetrahydroquinoline-1-carboxylate (29.6 g, 0.110 mol), pyridine (29.6 mL, 0.368 mol), potassium carbonate (30.5 g, 0.220 mol), methyl (1R,3R)-3-aminocyclohexane- 1 -carboxylate (25.6 g, 162.84 mmol) in DMSO (270 mL) was stirred for 15 h at 90 C and then cooled to room temperature. The reaction was quenched by the addition of water (200 mL) and extracted with ethyl acetate (3 x 300 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum.
The resulting crude product was purified by silica gel chromatography (eluting with 1:1 ethyl acetate/petroleum ether) to afford methyl (2S )-64R1R,3R)-3-(methoxycarbonyl)cyclohexyl] amino] -2-methyl-5-nitro- 1,2,3 ,4-tetrahydro quinoline-1-carboxylate as a red oil (32 g, 72%). LCMS (ES, m/z): 406 [M+H]t Step 7. methyl (2S)-5-amino-6-[[(1R,3R)-3-(methoxycarbonyl)cyclohexyl]amino]-2-methyl-1,2,3,4-tetrahydroquinoline-1-carboxylate [0130] A solution of methyl (2S )-64R1R,3R)-3-(methoxycarbonyl)cyclohexyl]amino]-2-methyl-5-nitro-1,2,3,4-tetrahydroquinoline- 1 -carboxylate (31.0 g, 76.46 mmol), NH4C1 (24.3 g, 454.28 mmol), Fe (64.3 g, 1.15 mol) in tetrahydrofuran (300 mL), ethanol (300 mL), and water (100 mL) was stirred for 1 h at 80 C and then cooled to room temperature. The solids were separated by filtration. The resulting solution was diluted with water (300 mL) and extracted with ethyl acetate (3 x 400 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to afford methyl (25)-5-amino-6- [R1R,3R)-3-(methoxycarbonyl)cyclohexyl] amino] -2-methyl- 1,2,3 ,4-tetrahydroquinoline-1-c arboxylate as a dark green solid (27.5 g, 92%). LCMS (ES, m/z): 376 [M+H]t Step 8. methyl (2S)-5-[2-(5-fluoro-2-methoxypheny1)-2-hydroxyacetamido]-6-[[(1R,3R)-3(methoxycarbonyl)cyclohexyllamino]-2-methyl-1,2,3,4-tetrahydroquinoline-1-carboxylate [0131]
A solution of 2-(5-fluoro-2-methoxypheny1)-2-hydroxyacetic acid (240 mg, 1.20 mmol), HATU (228 mg, 0.60 mmol), methyl (25)-5-amino-64R1R,3R)-3-(methoxycarbonyl)cyclohexyl] amino] -2-methyl- 1,2,3 ,4-tetrahydroquinoline-1-c arboxylate (150 mg, 0.40 mmol), DIEA (0.19 mL, 1.20 mmol) in N,N-dimethylformamide (10 mL) was stirred for 1 h at 25 C. The resulting solution was diluted with H20 (10 mL). The resulting solution was extracted with ethyl acetate (3x15 mL) and the organic layers combined. The resulting mixture was washed with brine (2x20 mL). The mixture was dried over anhydrous sodium sulfate and concentrated under vacuum. The resulting crude product was purified by silica gel chromatography (eluting with 3:2 ethyl acetate/petroleum ether) to afford methyl (2S)-542-(5fluoro-2-methoxypheny1)-2-hydroxyacetamido] -6- [ R1R,3R)-3 -(methoxyc arbonyl)c yclohexyl] amino] -2-methy1-1,2,3,4-tetrahydroquinoline-1-carboxylate as a yellow solid (180 mg, 81%). LCMS (ES, m/z): 558 [M+H]t Step 9. methyl (7S)-2-[(5-fluoro-2-methoxyphenyl)(hydroxy)methy1]-3-[(1R,3R)-3-(methoxycarbonyl)cyclohexyl]-7-methyl-3H,6H,7H,8H,9H-imidazo[4,5-flquinoline-6-carboxylate.
[0132] A solution of methyl (2S)-5-[2-(5-fluoro-2-methoxypheny1)-2-hydroxyacetamido]-6-[R1R,3R)-3-(methoxycarbonyl)cyclohexyl] amino] -2-methy1-1,2,3 ,4-tetrahydroquinoline- 1-carboxylate (180 mg, 0.32 mmol) in AcOH (8 mL) was stirred overnight at 60 C.
The reaction mixture was cooled and concentrated under vacuum. The resulting crude product was purified by silica gel chromatography (eluting with 1:1 ethyl acetate/petroleum ether) to afford methyl (7S)-2- R5-fluoro-2-methoxyphenyl)(hydroxy)methyl] -3- R1R,3R)-3 -(methoxyc arbonyl)c yclohexyl] -7-methyl-3H,6H,7H,8H,9H-imidazo[4,5-f]quinoline-6-carboxylate as a yellow solid (120 mg, 69%). LCMS (ES, m/z): 540 [M+H]t Step 10. (1R,3R)-3-[(7S)-2-[(R)-(5-fluoro-2-methoxyphenyl)(hydroxy)methyl]-6-(methoxycarbony1)- 7- methy1-3H,6H,7H,8H,9H- imidazo [4,5 - f] quinolin-3 -yl]
cyclohexane-1 -carboxylic acid (1); (1R,3R)-3-[(7S)-2-[(S)-(5-fluoro-2-methoxyphenyl)(hydroxy)methyl]-6-(methoxycarbony1)- 7- methyl-3H,6H,7H,8H,9H- imidazo [4,5 - f] quinolin-3 -yl]
cyclohexane-1 -carboxylic acid (1') [0133] A solution of methyl (7S)-24(5-fluoro-2-methoxyphenyl)(hydroxy)methyl]-3-R1R,3R)-3-(methoxycarbonyl)cyclohexyl] -7-methyl-3H,6H,7H,8H,9H-imidazo [4,5-f]
quinoline-6-carboxylate (120 mg, 0.22 mmol), and LiOH (16 mg, 0.67 mmol) in tetrahydrofuran (2.0 mL), methanol (2.0 mL) and water (2.0 mL) was stirred overnight at 25 C. The resulting mixture was concentrated under vacuum. The crude product was purified by Prep-HPLC
(Column, XBridge Prep C18 OBD Column, 19x150 mm, Sum; Mobile phase, A: water (containing 10 mmol/L
NH4HCO3) and B: ACN (15.0% to 29.0% over 14 min); Detector, UV 220/254nm). The product was separated by Chiral-Prep-HPLC (Column, CHIRALPAK IE, 2x25cm, 5 um; Mobile phase, A: Hex (containing 0.1%FA) and B: ethanol (hold 50.0% ethanol over 12 min);
Detector, UV

220/254 nm). The product fractions were concentrated to afford (1R,3R)-3-[(7S)-2-[(R)-(5-fluoro-2-methoxyphenyl)(hydroxy)methyll-6-(methoxycarbony1)-7-methyl-3H,6H,7H,8H,9H-imidazo[4,5-fiquinolin-3-yl[cyclohexane-l-carboxylic acid (1) as a white solid (23.6 mg, 20%);
and (1R,3R)-3 - [(7S )-2- RS )-(5-fluoro-2-methoxyphenyl)(hydroxy)methyll -6-(methoxyc arbony1)-7-methy1-3H,6H,7H,8H,9H-imidazo [4,5-f] quinolin-3 -y1] c yclohexane- 1-carboxylic acid (1') as a white solid (23.8 mg, 20%). Stereoisomeric purity was determined via HPLC:
Column:
CH1RALPAK 1E-3, Column size: 0.46 x 5 cm; 3 iim; Mobile phase: Hex (0.1%FA) :
Et0H =
50:50, Flow: 1.0 ml/min.
First eluting isomer (1): 1H-NMR (CD30D, 400 MHz) 6 (ppm): 7.56-7.47 (m, 1H), 7.47-7.31 (m, 1H), 7.21-7.09 (m, 1H), 7.09-6.89 (m, 2H), 6.53(s, 1H), 4.81-4.61(m, 2H), 3.85(s, 3H), 3.78(s, 3H), 3.31-3.18(m, 1H), 3.06-2.82 (m, 2H), 2.57-2.41 (m, 1H), 2.41-2.31 (m, 1H), 2.312.09 (m, 3H), 1.83-1.58 (m, 3H), 1.49-1.21 (m, 2H), 1.16 (d, J= 6.8 Hz, 3H). LCMS
(ES, m/z):
526 [M+H] .
Second eluting isomer (1'): 1H-NMR (CD30D, 400 MHz) 6 (ppm): 7.69-7.44 (m, 2H), 7.44-7.29 (m, 1H), 7.12-6.99 (m, 1H), 6.98-6.82 (m, 1H), 6.37(s, 1H), 5.03-4.91(m, 1H), 4.81-4.69(m, 1H), 3.78(s, 3H), 3.61(s, 3H), 3.22-3.04(m, 1H), 3.02-2.87 (m, 2H), 2.54-2.41 (m, 1H), 2.41-2.27 (m, 1H), 2.27-2.08 (m, 3H), 1.82-1.58 (m, 3H), 1.58-1.41 (m, 2H), 1.14 (d, J=
6.4 Hz, 3H).
LCMS (ES, m/z): 526 [M+H]t [0134] A composition of Formula (I) can comprise a compound of one or more of Formula (II-a), (II-b), (II-c), (II-d), (II-e), (II-f), (II-g), (II-h), (II-i), (II-j), (II-k), (II-1), (II-m), (II-n), and/or (II-o).
For example, in some embodiments the disclosure provides a composition comprising compound 1 of the foregoing structure or a pharmaceutically acceptable salt thereof at a purity of at least 90%
wherein the composition comprises less than 10%, e.g. less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2% or less than 1%, collectively of one or more of the following stereoisomers of compound 1, represented as Formulae (II-a) -(II-o) below:

OH OH OH
N¨

N N-...0 N N

fr¨OH in¨OH

I (II-a) o o I (II-b) o o I
(II-c) / / /

...,n0H
N¨ N¨

O H 1 N¨

N-...C1. N-...0 N N
.-- ¨OH N
t¨OH

----I (II-d) I (II-e) I
(II-f) OH OH OH
N-- N¨ N¨

N-...\3_ N-...0 _ OH l-- ¨OH )----¨OH

I (II-g) I (II-h) I
(II-i) /

-.HON
N¨

N¨ N¨

N-....0 N-....9...

)---OH

ri---- HO

I (II-j) I (II-k) I
(II-1) / o/ F 0/

"...OH
N=rr N---N-oun N

I (II-m) I (II-n) (I1-0)/compound 1' [0135] For instance, the disclosure provides a pharmaceutical composition comprising compound 1 or a pharmaceutically acceptable salt thereof at a purity of at least 95% as determined by the above HPLC method of Example 1.7. The disclosure also provides a pharmaceutical composition comprising compound 1 at a purity of at least 95% as determined by the above HPLC method.
[0136] The disclosure provides a compound of Formula II obtained by the foregoing method exemplified in Example 1.2:
/

OH
N ----N

I (II) or a pharmaceutically acceptable salt, enantiomer, hydrate, solvate, isomer or tautomer thereof.
[0137] It will be apparent to the skilled reader that each of the stereoisomers of the compound of Formula (II) can be obtained by varying the stereochemistry of the appropriate reagents utilized in the method of Example 1.2 above. For instance, by adjusting the reagent used in Step 4 of Example 1.2, compounds such as those of Formulae (II-m) and (II-n) can be synthesized.
Similarly, in Step 6 of Example 1.2, the reagent methyl (1S ,3R)-3-aminocyclohexane-l-carboxylate can be used in place of methyl (1R,3R)-3-aminocyclohexane- 1-carboxylate to obtain compounds of Formulae (II-b) and (II-e). It will be apparent to the skilled reader that by making a combination of these types of modifications to the process set out in Example 1.2, each of compounds (II-a) to (II-o) depicted above can be synthesized.

Example 1.3: (1R,3R)-3-[(78)-2-[(R)-hydroxy(phenyl)methyl]-6-(methoxycarbony1)-methyl-3H,6H,7H,8H,9H-imidazo[4,5-fiquinolin-3-yl]cyclohexane-1-carboxylic acid (2) [0138] Figure 2(C) provides a synthetic scheme for the preparation of Compound 2, as described below.
Step]. 6-fluoro-2-methyl-5-nitroquinoline [0139] A solution of trifluoromethanesulfonic acid (82.0 mL, 0.923 mol) in HNO3 (19.6 mL, 0.437 mol) was stirred for 20 min at 0 C. This was followed by the addition of 6-fluoro-2-methylquinoline (50.0 g, 0.310 mol) in dichloromethane (300 mL) at 0 C. The resulting mixture was stirred for 15 h at room temperature (25 C). The reaction mixture was diluted with water (300 mL). The pH value of the solution was adjusted to 8 with sodium bicarbonate (saturated aqueous solution). The resulting solution was extracted with dichloromethane (3 x 300 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum.
The residue was purified by silica gel chromatography (eluting with 1:4 ethyl acetate/petroleum ether) to afford 6-fluoro-2-methyl-5-nitroquinoline as a light yellow solid (60.0 g, 94%). LCMS
(ES, m/z): 207 [M+H]t Step 2. (28)-6-fluoro-2-methyl-5-nitro-1,2,3,4-tetrahydroquinoline [0140] A solution of (S)-(-)-Me0-BIPHEP (1.03 g, 1.77 mmol), chloro(1,5-cyclooctadiene)iridium(I) dimer (538 mg, 0.80 mmol) in toluene (100 mL) was stirred for 30 min at room temperature (25 C) under an atmosphere of nitrogen. This was followed by the addition of 12 (410 mg, 1.62 mmol), 6-fluoro-2-methyl-5-nitroquinoline (33.0 g, 0.160 mol) in toluene (100 mL). The resulting mixture was stirred for 20 h at room temperature (25 C) under hydrogen (50 atm). The resulting mixture was concentrated under vacuum and purified by silica gel chromatography (eluting with 1:1 ethyl acetate/petroleum ether) to afford the crude product (35.0 g). The crude product was dissolved in ethyl acetate (230 mL), followed by the addition of D-Camphorsulfonic acid (36.9 g, 0.158 mol). The resulting solution was stirred for 1 h at 60 C and then cooled to room temperature. The solids were collected by filtration, and rinsed with ethyl acetate (120 mL). The solids were dissolved in water (50 mL). The pH value of the solution was adjusted to 8 with sodium bicarbonate (saturated aqueous solution). The resulting solution was extracted with ethyl acetate (3 x 120 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to afford (2S)-6-fluoro-2-methy1-5-nitro-1,2,3,4-tetrahydroquinoline as a red solid (25.5 g, 76%). LCMS (ES, m/z): 211 [M+H]t Step 3. methyl (2S)-6-fluoro-2-methyl-5-nitro-1,2,3,4-tetrahydroquinoline-1-carboxylate [0141] A solution of (2S )-6-fluoro-2-methyl-5-nitro-1,2,3,4-tetrahydroquinoline (25.3 g, 0.120 mol), pyridine (39.0 mL, 0.484 mol), methyl carbonochloridate (18.7 mL, 0.242 mol) in dichloromethane (150 mL) was stirred for 3 h at room temperature (25 C). The reaction was washed with 1M hydrochloric acid (2 x 70 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to afford methyl (25)-6-fluoro-2-methy1-5-nitro-1,2,3,4-tetrahydroquinoline-1-carboxylate as a yellow solid (29.8 g, 92%).
LCMS (ES, m/z): 269 [M+H]t Step 4. methyl (2S)-6-[[(1R,3R)-3-(methoxycarbonyl)cyclohexyl]amino]-2-methyl-5-nitro-1,2,3,4-tetrahydroquinoline-1-carboxylate [0142] A solution of methyl (2S )-6-fluoro-2-methy1-5-nitro-1,2,3,4-tetrahydroquinoline-1-carboxylate (29.6 g, 0.110 mol), pyridine (29.6 mL, 0.368 mol), potassium carbonate (30.5 g, 0.220 mol), methyl (1R,3R)-3-aminocyclohexane- 1 -carboxylate (25.6 g, 162.84 mmol) in DMSO (270 mL) was stirred for 15 h at 90 C and then cooled to room temperature. The reaction was quenched by the addition of water (200 mL) and extracted with ethyl acetate (3 x 300 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum.
The resulting crude product was purified by silica gel chromatography (eluting with 1:1 ethyl acetate/petroleum ether) to afford methyl (25)-64R1R,3R)-3-(methoxycarbonyl)cyclohexyl] amino] -2-methyl-5-nitro- 1,2,3 ,4-tetrahydro quinoline-1-carboxylate as a red oil (32 g, 72%). LCMS (ES, m/z): 406 [M+H]t Step 5. methyl (2S)-5-amino-6-[[(1R,3R)-3-(methoxycarbonyl)cyclohexyl]amino]-2-methyl-1,2,3,4-tetrahydroquinoline-1-carboxylate 101431 A solution of (2S )-64R1R,3R)-3-(methoxycarbonyl)cyclohexyllamino]-2-methy1-5-nitro-1,2,3,4-tetrahydroquinoline- 1 -carboxylate (31.0 g, 76.46 mmol), NH4C1 (24.3 g, 454.28 mmol), Fe (64.3 g, 1.15 mol) in tetrahydrofuran (300 mL), ethanol (300 mL), water (100 mL) was stirred for 1 h at 80 C and then cooled to room temperature. The solids were filtered out by filtration.
The resulting solution was diluted with water (300 mL) and extracted with ethyl acetate (3 x 400 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to afford methyl (2S )-5-amino-6-[R1R,3R)-3 -(methoxycarbonyl)cyclohexyl] amino] -2-methyl- 1,2,3 ,4-tetrahydroquinoline-1-c arboxylate as a dark green solid (27.5 g, 92%). LCMS (ES, m/z): 376 [M+H]t Step 6. methyl (2S)-5-((R)-2-hydroxy-2-phenylacetamido)-6-[[(1R,3R)-3-(methoxycarbonyl)cyclohexyllamino]-2-methyl-1,2,3,4-tetrahydroquinoline-1-carboxylate 101441 A solution of (R)-2-hydroxy-2-phenylacetic acid (972 mg, 6.39 mmol), HATU (1.20 g, 3.16 mmol), methyl (2S )-5-amino-6- [R1R,3R)-3-(methoxycarbonyl)cyclohexyl]
amino] -2-methy1-1,2,3,4-tetrahydroquinoline-1-carboxylate (800 mg, 2.13 mmol), DIEA
(1.08 mL, 6.20 mmol) in N,N-dimethylformamide (10 mL) was stirred for 5 h at room temperature (25 C). The resulting solution was diluted with water (30 mL), and extracted with ethyl acetate (3 x 50 mL).
The organic layers were combined and washed with brine (2 x 25 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The resulting crude product was purified by silica gel chromatography (eluting with 1:1 ethyl acetate/petroleum ether) to afford methyl (25)-54(R)-2-hydroxy-2-phenylacetamido)-6-[R1R,3R)-3-(methoxycarbonyl)cyclohexyl] amino] -2-methy1-1,2,3 ,4-tetrahydroquinoline- 1-carboxylate as a colorless oil (600 mg, 55%). LCMS (ES, m/z): 510 [M+H]t Step 7. methyl (7S)-2-[(R)-hydroxy(phenyl)methy1]-3-[(1R,3R)-3-(methoxycarbonyl)cyclohexyl]-7-methyl-3H,6H,7H,8H,9H-imidazo[4,5-flquinoline-6-carboxylate [0145] A solution of methyl (2S)-5-((R)-2-hydroxy-2-phenylacetamido)-6-[R1R,3R)-3-(methoxycarbonyl)cyclohexyl] amino] -2-methyl- 1,2,3 ,4-tetrahydroquinoline-1-c arboxylate (600 mg, 1.18 mmol) in glacial acetic acid (5 mL, 98%) was stirred for overnight at 40 C and then cooled to room temperature. The reaction mixture was diluted with water (10 mL). The pH value of the solution was adjusted to 8 with sodium bicarbonate (saturated aqueous solution). The resulting solution was extracted with ethyl acetate (3 x 15 mL). The organic layers were combined and dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The resulting crude product was purified by silica gel chromatography (eluting with 1:1 ethyl acetate/petroleum ether) to afford methyl (7S )-2-RR)-hydroxy(phenyl)methyl] -3-R1R,3R)-3-(methoxycarbonyl)cyclohexyl] -7-methyl-3H,6H,7H,8H,9H-imidazo [4,5-f]
quinoline-6-carboxylate(400 mg, 69%) as a colorless oil. LCMS (ES, m/z): 492 [M+H]t Step 8. (1R,3R)-3-[(7S)-2-[(R)-hydroxy(phenyl)methyl]-6-(methoxycarbony1)-7-methyl-3H,6H,7H,8H,9H-imidazo-[4,541-quinolin-3-yl]cyclohexane-1-carboxylic acid (2) [0146] A solution of methyl (7S)-2-[(R)-hydroxy(phenyl)methy11-3-[(1R,3R)-3-(methoxycarbonyl)cyclohexyll -7-methyl-3H,6H,7H,8H,9H-imidazo [4,5-fl quinoline-6-carboxylate (400 mg, 0.81 mmol), LiOH (100 mg, 4.17 mmol) in tetrahydrofuran (5 mL) and water (2 mL) was stirred for overnight at room temperature (25 C). The resulting mixture was concentrated under vacuum. The crude product was purified by Prep-HPLC
(Column: XBridge Shield RP18 OBD Column, Sum, 19 x 150 mm; Mobile Phase, A: water (containing 10 mmol/L
NH4HCO3) and B: ACN (3% to 30% over 21 min); Detector: UV 254 nm). The product fractions were lyophilized to afford (1R,3R)-3-[(7S)-2-[(R)-hydroxy(phenyl)methyll-6-(methoxycarbony1)-7-methyl-3H,6H,7H,8H,9H-imidazo- [4,5-f] -quinolin-3-yl] c yclohexane- 1-carboxylic acid as a white solid (83.7 mg, 22%). Stereoisomeric purity was determined via HPLC:
Column: CHIRALPAK 1E-3, Column size: 0.46 x 5 cm; 3 iim; Mobile phase: Hex (0.1%FA):
Et0H = 85:15, Flow :1.0m1/min.
1H-NMR (CD30D, 400 MHz) 6 (ppm): 7.47-7.28 (m, 7H), 6.12(s, 1H), 4.84-4.74(m, 2H), 3.79(s, 3H), 3.33-3.25(m, 1H), 3.03-2.96 (m, 1H), 2.86-2.82 (m, 1H), 2.38-2.25 (m, 2H), 2.25-2.07 (m, 3H), 1.79-1.72 (m, 1H), 1.64-1.57 (m, 2H), 1.40-1.29 (m, 2H), 1.16 (d, J = 6.8 Hz, 3H). LCMS
(ES, m/z): 478 [M+H]; 99.13% ee.
[0147] A composition of Formula (I) can comprise a compound of one or more of Formula (M-a), (III-b), (III-c), (III-d), (III-e), (III-f), (III-g), (III-h), (III-i), (III-j), (III-k), (III-1), (III-m), (III-n), and/or (III-o). For example, in some embodiments the disclosure provides a composition comprising compound 2 of the foregoing structure or a pharmaceutically acceptable salt thereof at a purity of at least 90% wherein the composition comprises less than 10%, e.g.
less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2% or less than 1%, collectively of one or more of the following stereoisomers of compound 2, represented as Formulae (III-a) ¨ (III-o) below:

. . .
OH
OH OH N¨

N¨ N¨ N-...0 N , 0 2)-OH
¨OH OH

(III-( 111-a) I (III-b) c) =,10H .,10H
..10H
N¨ N¨ N¨

N N N
02)¨OH 0/2---OH

I (III-d) 1 (III-e) I
(III-0 OH OH OH
N¨ N¨ N¨

N-....C.,. N.õg N¨Q, 7.
o----OH

I (III-g) I (III-h) I
(III-i) . . .
-10H ..10H .,10H
N¨ N¨ N¨

N, N¨<?, 0 N-....0 N
02)-- ¨OH
crOH

I (III-j) I (III-k) 1 (III-1) ilfr . ilfr OH ..10H -10H
N-1\1,,Q
0)-OH 0' N OH N

0)-OH

I I (III-n) I (III-m) (III-o) [0148] For instance, the disclosure provides a pharmaceutical composition comprising compound 2 or a pharmaceutically acceptable salt thereof at a purity of at least 95% as determined by the above HPLC method of Example 1.7. The disclosure also provides a pharmaceutical composition comprising compound 2 at a purity of at least 95% as determined by the above HPLC method.
Example 1.4: (1R,3R)-3-[(7S)-2-[(S)-[2-(difluoromethoxy)-5-fluorophenyl](hydroxy) methy1]-6-(methoxycarbony1)-7-methyl-3H,6H,7H,8H,9H-imidazo[4,54]quinolin-3-yl]cyclohexane-1-carboxylic acid (452), (1R,3R)-3-[(7S)-2-[(R)-[2-(difluoromethoxy)-5-fluoropheny1](hydroxy)methy1]-6-(methoxycarbony1)-7-methyl-3H,6H,7H,8H,9H-imidazo [4,54]quinolin-3-yl]cyclohexane-1-carboxylic acid (3) [0149] Figure 2(D) provides a synthetic scheme for the preparation of Compound 3 and Compound 452, as described below.
Step]. 2-(difluoromethoxy)-5-fluorobenzaldehyde [0150] A solution of 5-fluoro-2-hydroxybenzaldehyde (2.0 g, 14.3 mmol), diethyl (bromodifluoromethyl)phosphonate (5.69 g, 21.3 mmol), potassium hydroxide (16.0 g, 285 mmol) in MeCN (100 mL) and water(50 mL) was stirred for 1 h at -30 C. The reaction mixture was diluted with water (20 mL). The resulting solution was extracted with ethyl acetate (3x100 mL) and the organic layers combined and dried over anhydrous sodium sulfate. The solids were filtered out. The resulting mixture was concentrated under vacuum. The resulting crude product was purified by silica gel chromatography (eluting with 1:1 ethyl acetate/petroleum ether) to afford 2-(difluoromethoxy)-5-fluorobenzaldehyde as a yellow solid (1.46 g, 54%). LCMS
(ES, m/z): 191 [M+H] .
Step 2. 2-[2-(difluoromethoxy)-5-fluoropheny1]-2-[(trimethylsilypoxy]acetonitrile [0151] A solution of 2-(difluoromethoxy)-5-fluorobenzaldehyde (1.46 g, 7.68 mmol), TMSCN
(760 mg, 7.66 mmol), ZnI2 (50 mg, 0.16 mmol) in dichloromethane (3 mL) was stirred for 2 h at room temperature (25 C). The resulting mixture was concentrated under vacuum.
The resulting crude product was purified by silica gel chromatography (eluting with 1:1 ethyl acetate/petroleum ether) to afford 2[2-(difluoromethoxy)-5-fluoropheny1]-2-[(trimethylsily1) oxy]acetonitrile as a yellow solid (800 mg, 36%) . LCMS (ES, m/z):290 [M+H]
Step 3. 242-(difluoromethoxy)-5-fluorophenyl]-2-hydroxyacetic acid [0152] A solution of 2-[2-(difluoromethoxy)-5-fluoropheny1]-2-[(trimethylsily1)oxy] acetonitrile (800 mg, 2.77 mmol), 1,4-dioxane (2.0 mL), hydrogen chloride (1.0 mL, 12M) in water (2 mL) was stirred for 12 h at 70 C and then cooled to room temperature. The resulting solution was concentrated under vacuum. The crude product was purified by reverse phase column chromatography (water (containing 0.05%TFA)/MeCN) to afford 242-(difluoromethoxy)-5-fluoropheny1]-2-hydroxyacetic acid (400 mg, 61%). LCMS (ES, m/z): 237 [M+H]t Step 4. 6-fluoro-2-methyl-5-nitroquinoline [0153] A solution of trifluoromethanesulfonic acid (82.0 mL, 0.923 mol) in HNO3 (19.6 mL, 0.437 mol) was stirred for 20 min at 0 C. This was followed by the addition of 6-fluoro-2-methylquinoline (50.0 g, 0.310 mol) in dichloromethane (300 mL) at 0 C. The resulting mixture was stirred for 15 h at room temperature (25 C). The reaction mixture was diluted with water (300 mL). The pH value of the solution was adjusted to 8 with sodium bicarbonate (saturated aqueous solution). The resulting solution was extracted with dichloromethane (3 x 300 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum.
The residue was purified by silica gel chromatography (eluting with 1:4 ethyl acetate/petroleum ether) to afford 6-fluoro-2-methyl-5-nitroquinoline as a light yellow solid (60.0 g, 94%). LCMS
(ES, m/z): 207 [M+H]t Step 5. (2S)-6-fluoro-2-methy1-5-nitro-1,2,3,4-tetrahydroquinoline [0154] A solution of (S)-(-)-Me0-BIPHEP (1.03 g, 1.77 mmol), chloro(1,5-cyclooctadiene)iridium(I) dimer (538 mg, 0.80 mmol) in toluene (100 mL) was stirred for 30 min at room temperature (25 C) under an atmosphere of nitrogen. This was followed by the addition of 12 (410 mg, 1.62 mmol), 6-fluoro-2-methyl-5-nitroquinoline (33.0 g, 0.160 mol) in toluene (100 mL). The resulting mixture was stirred for 20 h at room temperature (25 C) under hydrogen (50 atm). The resulting mixture was concentrated under vacuum and purified by silica gel chromatography (eluting with 1:1 ethyl acetate/petroleum ether) to afford the crude product (35.0 g). The crude product was dissolved in ethyl acetate (230 mL), followed by the addition of D-Camphorsulfonic acid (36.9 g, 0.158 mol). The resulting solution was stirred for 1 h at 60 C and then cooled to room temperature. The solids were collected by filtration, and rinsed with ethyl acetate (120 mL). The solids were dissolved in water (50 mL). The pH value of the solution was adjusted to 8 with sodium bicarbonate (saturated aqueous solution). The resulting solution was extracted with ethyl acetate (3 x 120 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to afford (2S)-6-fluoro-2-methy1-5-nitro-1,2,3,4-tetrahydroquinoline as a red solid (25.5 g, 76%). LCMS (ES, m/z): 211 [M+H]t Step 6. methyl (2S)-6-fluoro-2-methyl-5-nitro-1,2,3,4-tetrahydroquinoline-1-carboxylate [0155] A solution of (2S )-6-fluoro-2-methyl-5-nitro-1,2,3,4-tetrahydroquinoline (25.3 g, 0.120 mol), pyridine (39.0 mL, 0.484 mol), methyl carbonochloridate (18.7 mL, 0.242 mol) in dichloromethane (150 mL) was stirred for 3 h at room temperature (25 C). The reaction was washed with 1M hydrogen chloride (2 x 70 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to afford methyl (25)-6-fluoro-2-methy1-5-nitro-1,2,3,4-tetrahydroquinoline-1-carboxylate as a yellow solid (29.8 g, 92%).
LCMS (ES, m/z): 269 [M+H]t Step 7. methyl (2S)-6-[[(1R,3R)-3-(methoxycarbonyl)cyclohexyl]amino]-2-methyl-5-nitro-1,2,3,4-tetrahydroquinoline-1-carboxylate [0156] A solution of methyl (2S )-6-fluoro-2-methy1-5-nitro-1,2,3,4-tetrahydroquinoline-1-carboxylate (29.6 g, 0.110 mol), pyridine (29.6 mL, 0.368 mol), potassium carbonate (30.5 g, 0.220 mol), methyl (1R,3R)-3-aminocyclohexane- 1 -carboxylate (25.6 g, 162.84 mmol) in DMSO (270 mL) was stirred for 15 h at 90 C and then cooled to room temperature. The reaction was quenched by the addition of water (200 mL) and extracted with ethyl acetate (3 x 300 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum.
The resulting crude product was purified by silica gel chromatography (eluting with 1:1 ethyl acetate/petroleum ether) to afford methyl (2S )-6- [R1R,3R)-3-(methoxycarbonyl)cyclohexyl]
amino]-2-methy1-5-nitro-1,2,3,4-tetrahydroquinoline-l-carboxylate as a red oil (32 g, 72%).
LCMS (ES, m/z): 406 [M+H]t Step 8. methyl (2S)-5-amino-6-[[(1R,3R)-3-(methoxycarbonyl)cyclohexyl]amino]-2-methyl-1,2,3,4-tetrahydroquinoline-1-carboxylate [0157] A solution of methyl (2S )-64R1R,3R)-3-(methoxycarbonyl)cyclohexyl]
amino]-2-methyl-5-nitro-1,2,3,4-tetrahydroquinoline- 1 -carboxylate (31.0 g, 76.46 mmol), NH4C1 (24.3 g, 454.28 mmol), Fe (64.3 g, 1.15 mol) in tetrahydrofuran (300 mL), ethanol (300 mL), water (100 mL) was stirred for 1 h at 80 C and then cooled to room temperature. The solids were filtered out by filtration. The resulting solution was diluted with water (300 mL) and extracted with ethyl acetate (3 x 400 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to afford methyl (2S )-5-amino-6- [R1R,3R)-3-(methoxycarbonyl)cyclohexyl] amino] -2-methyl- 1,2,3 ,4-tetrahydroquinoline-1-c arboxylate as a dark green solid (27.5 g, 92%). LCMS (ES, m/z): 376 [M+H]t Step 9. methyl (28)-542-[2-(difluoromethoxy)-5-fluoropheny1]-2-hydroxyacetamido]-6-[[(1R,3R)-3-(methoxycarbonyl)cyclohexyllamino]-2-methyl-1,2,3,4-tetrahydroquinoline-1-carboxylate [0158] A solution of methyl (2S )-5-amino-6-[[(1R,3R)-3-(methoxycarbonyl)cyclohexyl] amino]-2-methy1-1,2,3,4-tetrahydroquinoline-l-carboxylate (200 mg, 0.53 mmol), 2- [2-(difluoromethoxy)-5-fluoropheny1]-2-hydroxyacetic acid (220 mg, 0.93 mmol), DMTMM (350 mg, 1.26 mmol) in dichloromethane (5 mL) was stirred for 1 h room temperature (25 C). The resulting solution was concentrated under vacuum. The resulting crude product was purified by silica gel chromatography (eluting with 1:1 ethyl acetate/petroleum ether) to afford methyl (2S)-5- [2- [2-(difluoromethoxy)-5-fluoropheny1]-2-hydroxyacetamido]-6- [R1R,3R)-3-(methoxycarbonyl)cyclohexyl] amino] -2-methyl- 1,2,3 ,4-tetrahydroquinoline-1-c arboxylate as a yellow solid (70.0 mg, 22%). LCMS (ES, m/z): 594 [M+H]t Step 10. methyl (78)-2-[[2-(difluoromethoxy)-5-fluorophenyl](hydroxy)methy1]-3-[(1R,3R)-3-(methoxycarbonyl)cyclohexyl]-7-methyl-3H,6H,7H,8H,9H-imidazo[4,5-flquinoline-carboxylate [0159] A solution of methyl (2S )-5- [2- [2-(difluoromethoxy)-5 -fluorophenyl] -2-hydroxyacetamido] -6- [R1R,3R)-3-(methoxycarbonyl)cyclohexyl] amino] -2-methy1-1,2,3,4-tetrahydroquinoline-l-carboxylate (70.0 mg, 0.12 mmol) in glacial acetic acid (2.0 mL) was stirred for overnight at 40 C and then cooled to room temperature. The resulting solution was concentrated under vacuum.
The resulting crude product was purified by silica gel chromatography (eluting with 1:2 ethyl acetate/petroleum ether) to afford methyl (75)-24[2-(difluoromethoxy)-5-fluorophenyl] (hydroxy)methyl] -3- R1R,3R)-3-(methoxycarbonyl) cyclohexyl]-7-methy1-3H,6H,7H,8H,9H-imidazo[4,5-fiquinoline-6-carboxylate as a yellow solid (50.0 mg, 74%). LCMS (ES, m/z): 576 [M+H]t Step 11. (1R,3R)-3-[(7S)-2-[(S)-[2-(difluoromethoxy)-5-fluorophenyl](hydroxy)methy1]-6-(methoxycarbony1)- 7-methy1-3H,6H,7H,8H,9H- imidazo [4,54] quinolin-3-yl]
cyclohexane-1 -carboxylic acid (452); (1R,3R)-3- [(7S)-2-[(R)- [2-(difluoromethoxy)-5-fluorophenyl](hydroxy)methy1]-6-(methoxycarbony1)-7-methyl-3H,6H,7H,8H,9H-imidazo [4,54] quinolin-3-yl] cyclohexane-1 -carboxylic acid (3) [0160] A solution of methyl (7S)-2-[[2-(difluoromethoxy)-5-fluorophenyl](hydroxy)methy1]-3-[(1R,3R)-3-(methoxycarbonyl)cyclohexyl] -7-methyl-3H,6H,7H,8H,9H-imidazo [4,5-f] quinoline -6-carboxylate (50.0 mg, 0.09 mmol), LiOH (10.0 mg, 0.42 mmol) in tetrahydrofuran (2.0 mL) and water (2.0 mL) was stirred for overnight at room temperature (25 C). The resulting mixture was concentrated under vacuum. The crude product was purified by Prep-HPLC
(Column, XBridge Shield RP18 OBD Column, 30x150 mm, 5 um; Mobile phase, A: water (containing 10 mmol/L NH4HCO3) and B: ACN (25.0% to 35.0% over 8 min); Detector, UV 254/220 nm). The product fractions were concentrated to afford (1R,3R)-3-[(7S)-2-[(S)-[2-(difluoromethoxy)-5-fluorophenyl](hydroxy)methyl] -6-(methoxyc arbony1)-7-methy1-3H,6H,7H,8H,9H-imidazo [4,5-f]quinolin-3-yl]cyclohexane- 1-carboxylic acid (452) as a white solid (4.50 mg, 9%), and (1R,3R)-3 - [(7S )-2- [(R)- [2-(difluoromethoxy)-5-fluorophenyl] (hydroxy)methyl] -6-(methoxyc arbony1)-7 -methyl-3H,6H,7H,8H,9H-imidazo [4,5-f] quinolin-3 -yl] c yclohexane- 1-carboxylic acid (515) as a white solid (4.30 mg, 9%). Enantiomeric excess was determined via HPLC:
Column:
CH1RALPAK 1E-3, Column size: 0.46 x 5 cm; 3 iim; Co-Solvent: IPA (20 mM NH3) Gradient (B%) : 10% to 50% in 4.0min, hold 2.0 min at 50%.
First eluting isomer (452): 1H-NMR (CD30D, 400 MHz) 6 (ppm): 7.63-7.61 (m, 1H), 7.53 (d, J
= 8.8 Hz, 1H), 7.41(d, J= 9.2Hz, 1H) 7.20-7.13 (m, 2H), 6.67-6.30 (m, 2H), 4.98-4.95 (m, 1H), 4.76-4.71 (m, 1H), 3.78 (s, 3H), 3.15-2.86 (m, 3H), 2.46-2.20 (m, 5H), 1.81-1.53 (m, 5H), 1.13 (d, J= 6.8 Hz, 3H). LCMS (ES, m/z): 562 [M+H]t Second eluting isomer (3): 1H-NMR (CD30D, 400 MHz) 6 (ppm): 7.55-7.53 (m, 1H), 7.47-7.42 (m, 2H), 7.40-7.12 (m, 2H), 6.85-6.44 (m, 2H), 4.94-4.91 (m, 1H), 4.76-4.71 (m, 1H), 3.78 (s, 3H), 3.22-2.84 (m, 3H), 2.46-2.23 (m, 5H), 1.84-1.61 (m, 5H), 1.14 (d, J = 6.4 Hz, 3H). LCMS (ES, m/z): 562 [M+H[ ; >99.99% ee.
[0161] In some embodiments, the disclosure provides the first eluting isomer obtained from Step 11 of the process described in Example 1.4. In some embodiments, the disclosure provides the second eluting isomer obtained from Step 11 of the process described in Example 1.4.

[0162] A composition of Formula (I) can comprise a compound of one or more of Formula (IV-a), (IV-b), (IV-c), (IV-d), (IV-e), (IV-f), (IV-g), (IV-h), (IV-i), (IV-j), (IV-k), (IV-1), (IV-m), (IV-n), and/or (IV-o). For example, in some embodiments the disclosure provides a composition comprising compound 3 of the foregoing structure or a pharmaceutically acceptable salt thereof at a purity of at least 90% wherein the composition comprises less than 10%, e.g.
less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2% or less than 1%, collectively of one or more of the following stereoisomers of compound 3, represented as Formulae (IV-a) ¨ (IV-o) below:
F F
F 40 0)¨F F . 0)¨F F 441 F0)¨F
OH OH OH
N¨

N-..<3..
N."0 N-.C7 101 , N N ==;- N

OH
ofp-OH0.70H----I (IV-a) I (IV-b) I
(IV-c) F
F 40 0 F 4. 0)_F F 40 0 -10H ..10H .,10H
N79. OH 0 OH N-...,C7 i N N "--- N

ol)--OH

I (IV-d) I (IV-e) I
(IV-f) F F F
F . 0 F 411 0 F 41 0 OH OH OH
N¨

N-.C....
..

OH
o;)-- 0 OH OH

I (IV-g) I (IV-h) I
(IV-i) F F F
F * 0 >-F
F * 0 )-F
F * 0 >-F
-10H ..10H -10H
N-N-.<1._ ..

OH
o2-- - 0 OH
---OH

I (IV-j) I (IV-k) I (IV-1) F F
F)-F F * 0)-F F afr 0 F afr 0)-F
OH -10H ..10H
N-N,,,9_ 0 ' N \`µµ. N la N
1.0 OH

I (IV-m) I (IV-n) I (IV-o) [0163] For instance, the disclosure provides a pharmaceutical composition comprising compound 3 or a pharmaceutically acceptable salt thereof at a purity of at least 95% as determined by the above HPLC method of Example 1.7. The disclosure also provides a pharmaceutical composition comprising compound 3 at a purity of at least 95% as determined by the above HPLC method.
Example 1.5: (1R,3R)-34(S)-2-benzy1-6-(methoxycarbony1)-7-methyl-6,7,8,9-tetrahydro-3H-imidazo[4,5-fiquinolin-3-y1)cyclohexane-1-carboxylic acid (4) [0164] Figure 2(E) provides a synthetic scheme for the preparation of Compound 4, as described below.
Step 1. methyl (S)-5-amino-6-(41R,3R)-3-(methoxycarbonyl)cyclohexyl)amino)-2-methyl-3,4-dihydroquinoline-1-(2H)-carboxylate [0165] Into a 10-mL round-bottom flask purged and maintained with an inert atmosphere of nitrogen, methyl (1R,3R)-3-aminocyclohexane-1-carboxylate hydrochloride (130 mg, 0.67 mmol) was dissolved in dioxane (4 mL). Then methyl (2S)-5-amino-6-bromo-2-methy1-1,2,3,4-tetrahydroquinoline-l-carboxylate (100 mg, 0.33 mmol, Intermediate 1), Brettphos (72 mg, 0.13 mmol), 3rd Generation Brettphos precatalyst (61 mg, 0.07 mmol) and sodium tert-butoxide (97 mg, 1.01 mmol) were added. The resulting solution was stirred for 1 h at 100 C under nitrogen atmosphere. The reaction mixture was cooled and the solids were filtered out.
The resulting mixture was concentrated under vacuum. The residue was subjected to purification by FCC eluting with ethyl acetate/petroleum ether (2:1). This afforded the title compound (41.3 mg, 33%) as a dark green solid. MS: (ES, m/z): 376 [M+H]t Step 2. methyl (S)-2-benzy1-3-41R,3R)-3-(methoxycarbonyl)cyclohexyl)-7-methyl-3,7,8,9-tetrahydro-6H-imidazo[4,5-fiquinoline-6-carboxylate [0166] Into a 25-mL round-bottom flask, methyl methyl (S)-5-amino-6-(((lR,3R)-(methoxyc arbonyl)c yclohexyl)amino)-2-methy1-3 ,4-dihydroquinoline- 1(2H)-c arboxylate (165.4 mg, 0.44 mmol) was dissolved in dichloromethane (5 mL). Then 2-phenylacetaldehyde (158.8 mg, 1.32 mmol) was added. The resulting solution was stirred for 2 h at room temperature. The resulting mixture was concentrated under vacuum. The residue was subjected to purification by FCC eluting with ethyl acetate/petroleum ether (2:1). This afforded the title compound (122.9 mg, 59%) as a yellow solid. MS: (ES, m/z): 476 [M+H]t Step 3.
(1R,3R)-3-[(78)-2-benzy1-6-(methoxycarbony1)-7-methyl-3H,6H,7H,8H,9H-imidazo [4,5-f] quinolin-3-yl] cyclohexane-1 -carboxylic acid (4) [0167] Into a 25-mL round-bottom flask, methyl (S)-2-benzy1-34(1R,3R)-3-(methoxycarbonyl)cyclohexyl)-7-methy1-3,7,8,9-tetrahydro-6H-imidazo [4,5-f]
quinoline-6-carboxylate (30 mg, 0.06 mmol) was dissolved in tetrahydrofuran (0.5 mL). Then water (0.5 mL) was added, followed by lithium hydroxide (7.0 mg, 0.29 mmol). The resulting solution was stirred for 3 h at 85 C. The pH value of the solution was adjusted to 5-6 with hydrochloric acid (1 mol/L).
The resulting solution was extracted with ethyl acetate (3x20 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The crude product was purified by Prep-HPLC with the following conditions: Column, XBridge C18 OBD Prep Column, 100A, 5 iim, 19 mm X 250 mm; mobile phase, A: Water (containing 10 mmol/L NH4HCO3) and B: ACN (10.0% to 30.0% ACN over 10 min); UV Detector:
254nm. This afforded the title compound (15.2 mg, 52%) as a white solid.
1H NMR (CD30D, 400 MHz) 6 (ppm): 7.47 (d, J = 9.0 Hz, 1H), 7.39 (d, J = 8.9 Hz, 1H), 7.35-7.19 (m, 5H), 4.84-4.68 (m, 2H), 4.45-4.25 (m, 2H), 3.79 (s, 3H), 3.22-3.14 (m, 1H), 2.98-2.85 (m, 2H), 2.40-2.02 (m, 5H), 1.83-1.70 (m, 1H), 1.64-1.54 (m, 2H), 1.33-1.13 (m, 5H). MS: (ES, m/z): 462 [M+H]t [0168] Compound 17, Compound 18, and Compound 19 were prepared using standard chemical manipulations and procedures similar to those used for the preparation of Example 16.

Example 1.6: 3-((7S)-2-((4-chlorophenyl)(hydroxy)methyl)-6-(methoxycarbonyl)-7-methyl-6,7,8,9-tetrahydro-3H-imidazo[4,5-fiquinolin-3-y1)cyclohexane-1-carboxylic acid (413); and (1R,3R)-34(S)-2-((S)-(4-chlorophenyl)(hydroxy)methyl)-6-(methoxycarbonyl)-7-methyl-6,7,8,9-tetrahydro-3H-imidazo[4,5-fiquinolin-3-y1)cyclohexane-1-carboxylic acid (501) [0169] Figure 2(F) provides a synthetic scheme for the preparation of Compound 413 and Compound 501, as described below.
Step]. 6-fluoro-2-methyl-5-nitroquinoline [0170] A solution of trifluoromethanesulfonic acid (82.0 mL, 0.923 mol) in HNO3 (19.6 mL, 0.437 mol) was stirred for 20 min at 0 C. This was followed by the addition of 6-fluoro-2-methylquinoline (50.0 g, 0.310 mol) in dichloromethane (300 mL) at 0 C. The resulting mixture was stirred for 15 hours at room temperature (25 C). The reaction mixture was diluted with water (300 mL). The pH value of the solution was adjusted to 8 with sodium bicarbonate (saturated aqueous solution). The resulting solution was extracted with dichloromethane (3 x 300 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (eluting with 1:4 ethyl acetate/petroleum ether) to afford 6-fluoro-2-methyl-5-nitroquinoline as a light yellow solid (60.0 g, 94%). LCMS (ES, m/z): 207 [M+H]t Step 2. (2S)-6-fluoro-2-methyl-5-nitro-1,2,3,4-tetrahydroquinoline [0171] A solution of (S)-(-)-Me0-BIPHEP (1.03 g, 1.77 mmol), chloro(1,5-cyclooctadiene)iridium(I) dimer (538 mg, 0.80 mmol) in toluene (100 mL) was stirred for 30 min at room temperature (25 C) under an atmosphere of nitrogen. This was followed by the addition of 12 (410 mg, 1.62 mmol), and 6-fluoro-2-methyl-5-nitroquinoline (33.0 g, 0.160 mol) in toluene (100 mL). The resulting mixture was stirred for 20 h at room temperature (25 C) under hydrogen (50 atm). The resulting mixture was concentrated under vacuum and purified by silica gel chromatography (eluting with 1:1 ethyl acetate/petroleum ether) to afford the crude product (35.0 g). The crude product was dissolved in ethyl acetate (230 mL), followed by the addition of D-Camphorsulfonic acid (36.9 g, 0.158 mol). The resulting solution was stirred for 1 h at 60 C and then cooled to room temperature. The solids were collected by filtration, and rinsed with ethyl acetate (120 mL). The solids were dissolved in water (50 mL). The pH value of the solution was adjusted to 8 with sodium bicarbonate (saturated aqueous solution). The resulting solution was extracted with ethyl acetate (3 x 120 mL). The combined organic layers was dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to afford (2S)-6-fluoro-2-methy1-5-nitro-1,2,3,4-tetrahydroquinoline as a red solid (25.5 g, 76%). LCMS (ES, m/z): 211 [M+H]t Step 3. methyl (2S)-6-fluoro-2-methyl-5-nitro-1,2,3,4-tetrahydroquinoline-1-carboxylate [0172] A solution of (2S )-6-fluoro-2-methyl-5-nitro-1,2,3,4-tetrahydroquinoline (25.3 g, 0.120 mol), pyridine (39.0 mL, 0.484 mol), and methyl carbonochloridate (18.7 mL, 0.242 mol) in dichloromethane (150 mL) was stirred for 3 h at room temperature (25 C). The reaction was washed with 1N hydrogen chloride (aq., 2 x 70 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to afford methyl (25)-6-fluoro-2-methy1-5-nitro-1,2,3,4-tetrahydroquinoline-1-carboxylate as a yellow solid (29.8 g, 92%).
LCMS (ES, m/z): 269 [M+H]t Step 4. methyl (2S)-6-[[(1R,3R)-3-(methoxycarbonyl)cyclohexyl]amino]-2-methyl-5-nitro-1,2,3,4-tetrahydroquinoline-1-carboxylate [0173] A solution of methyl (2S )-6-fluoro-2-methy1-5-nitro-1,2,3,4-tetrahydroquinoline-1-carboxylate (29.6 g, 0.110 mol), pyridine (29.6 mL, 0.368 mol), potassium carbonate (30.5 g, 0.220 mol), and methyl (1R,3R)-3-aminocyclohexane- 1 -carboxylate (25.6 g, 162.84 mmol) in DMSO
(270 mL) was stirred for 15 h at 90 C and then cooled to room temperature.
The reaction was quenched by the addition of water (200 mL) and extracted with ethyl acetate (3 x 300 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The resulting crude product was purified by silica gel chromatography (eluting with 1:1 ethyl acetate/petroleum ether) to afford methyl (25)-6-[R1R,3R)-3-(methoxycarbonyl)cyclohexyl] amino] -2-methyl-5-nitro- 1,2,3 ,4-tetrahydro quinoline-1-carboxylate as a red oil (32 g, 72%). LCMS (ES, m/z): 406 [M+H]t Step 5. methyl (2S)-5-amino-6-[[(1R,3R)-3-(methoxycarbonyl)cyclohexyl]amino]-2-methyl-1,2,3,4-tetrahydroquinoline-1-carboxylate 101741 A solution of methyl (2S )-2-methy1-5-nitro-64R1R,3R)-4-(methoxycarbonyl)cyclohexyl] amino] -1,2,3 ,4-tetrahydroquinoline- 1 -carboxylate (31.0 g, 76.46 mmol), NH4C1 (24.3 g, 454.28 mmol), and Fe (powder, 64.3 g, 1.15 mol) in tetrahydrofuran (300 mL), ethanol (300 mL), water (100 mL) was stirred for 1 h at 80 C and then cooled to room temperature. The solids were filtered out by filtration. The resulting solution was diluted with water (300 mL) and extracted with ethyl acetate (3 x 400 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to afford methyl (2S )-5-((R)-2-hydroxy-2-phenylacetamido)-6- [R1R,3R)-3-(methoxycarbonyl)cyclohexyl]
amino]-2-methy1-1,2,3,4-tetrahydroquinoline-1-carboxylate as a dark green solid (27.5 g, 92%).
LCMS (ES, m/z): 376 [M+H]t Step 6. methyl (2S)-542-(4-chloropheny1)-2-hydroxyacetamido]-6-[[(1R,3R)-3-(methoxycarbonyl)cyclohexyliamino]-2-methyl-1,2,3,4-tetrahydroquinoline-1-carboxylate 101751 A solution of 2-(4-chloropheny1)-2-hydroxyacetic acid (112 mg, 0.60 mmol), HATU (304 mg, 0.80 mmol), methyl (25 )-5- amino-6- [R1R,3R)-3-(methoxycarbonyl)cyclohexyl] amino] -2-methy1-1,2,3,4-tetrahydroquinoline-1-carboxylate (150 mg, 0.40 mmol), and DlEA
(155 mg, 1.20 mmol) in N,N-dimethylformamide (2 mL) was stirred for 15 h at room temperature (25 C). The resulting solution was diluted with water (30 mL), and extracted with ethyl acetate (3 x 50 mL).
The organic layers were combined and washed with brine (2 x 25 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The resulting crude product was purified by silica gel chromatography (eluting with 1:1 ethyl acetate/petroleum ether) to afford methyl (2S)-5-[2-(4-chloropheny1)-2-hydroxyacetamido]-6-[R1R,3R)-3-(methoxycarbonyl)cyclohexyl] amino] -2-methy1-1,2,3 ,4-tetrahydroquinoline- 1-carboxylate as yellow oil (70.0 mg, 32%). LCMS (ES, m/z): 544 [M+H]t Step 7. methyl (7S)-2-[(4-chlorophenyl)(hydroxy)methy11-3-[(1R,3R)-3-(methoxycarbonyl)cyclohexy11-7-methyl-3H,6H,7H,8H,9H-imidazo[4,5-flquinoline-6-carboxylate [01176]
A solution of methyl (2S)-5- [2-(4-chloropheny1)-2-hydroxyacetamido]-6-[R1R,3R)-3 -(methoxyc arbonyl)c yclohexyl] amino] -2-methy1-1,2,3 ,4-tetrahydroquinoline- 1-carboxylate (60.0 mg, 0.11 mmol) in AcOH (2 mL) was stirred for 15 h at 40 C
and then cooled to room temperature. The reaction mixture was diluted with water (10 mL). The pH value of the solution was adjusted to 8 with sodium bicarbonate (saturated aqueous solution). The resulting solution was extracted with ethyl acetate (3 x 15 mL). The organic layers were combined and dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The resulting crude product was purified by silica gel chromatography (eluting with 1:1 ethyl acetate/petroleum ether) to afford methyl (7S )-2-R4-chlorophenyl)(hydroxy)methyl] -3- R1R,3R)-3-(methoxycarbonyl)cyclohexyl] -7-methyl-3H,6H,7H,8H,9H-imidazo [4,5-f]
quinoline-6-carboxylate as yellow oil (46.0 mg, 79%). LCMS (ES, m/z): 526 [M+H]t Step 8. (1R,3R)-3-[(7S)-2-[(R)-(4-chlorophenyl)(hydroxy)methyl]-6-(methoxycarbony1)-7-methy1-3H,6H,7H,8H,9H-imidazo[4,5-fiquinolin-3-yl]cyclohexane-1-carboxylic acid (413);
(1R,3R)-3-[(7S)-2-[(S)-(4-chlorophenyl)(hydroxy)methyl]-6-(methoxycarbony1)-7-methy1-3H,6H,7H,8H,9H-imidazo[4,5-fiquinolin-3-yl]cyclohexane-1-carboxylic acid (501) [0177] A solution of methyl (7S)-2-[(4-chlorophenyl)(hydroxy)methyll -3-[(1R,3R)-3-(methoxyc arbonyl)c yclohexyl] -7-methyl-3H,6H,7H,8H,9H-imidazo [4,5-f]
quinoline-6-carboxylate (50.0 mg, 0.10 mmol), and LiOH (11.4 mg, 0.48 mmol) in tetrahydrofuran (1 mL) and water (1 mL) was stirred for 15 h at 25 C. The resulting mixture was concentrated under vacuum.
The crude product was purified by Prep-HPLC (Column: XBridge Shield RP18 OBD
Column, Sum, 19 x 150 mm; Mobile Phase, A: water (containing 10 mmol/L NH4HCO3) and B:
ACN (10%
to 37% over 12 min); Detector: UV 254 nm). The product fractions were lyophilized to afford (1R,3R)-3-[(7S)-2-[(R)-(4-chlorophenyl)(hydroxy)methyl] -6-(methoxyc arb ony1)-7 -methyl-3H,6H,7H,8H,9H-imidazo[4,5-fiquinolin-3-yl[cyclohexane- 1-carboxylic acid (413) as a white solid (10.5 mg, 43%); and (1R,3R)-3-[(75)-2-[(S)-(4-chlorophenyl)(hydroxy)methyll-6-(methoxyc arbony1)-7-methy1-3H,6H,7H,8H,9H-imidazo [4,5-f] quinolin-3 -yl] c yclohexane- 1-carboxylic acid (501) as a white solid (7.0 mg, 29%).
First eluting isomer (413): 1H-NMR (CD30D, 400 MHz) 6 (ppm): 7.49 (d, J= 9.0 Hz, 1H), 7.42-7.33 (m, 5H), 6.19 (s, 1H), 4.92-4.90 (m, 1H), 4.82-4.72 (m, 1H), 3.79 (s, 3H), 3.34-3.20 (m, 1H), 3.02-2.94 (m, 1H), 2.90-2.87 (m, 1H), 2.36-2.09 (m, 4H), 1.99-1.96 (m, 1H), 1.80-1.42 (m, 5H), 1.16 (d, J= 6.6 Hz, 3H). LCMS (ES, m/z): 512 [M+H]t Second eluting isomer (501): 1H-NMR (CD30D, 400 MHz) 6 (ppm): 7.52-7.33 (m, 6H), 6.22 (s, 1H), 4.84-4.73 (m, 2H), 3.78 (s, 3H), 3.27-3.16 (m, 1H), 3.04-2.92 (m, 1H), 2.90-2.88 (m, 1H), 2.46-2.35 (m, 2H), 2.30-2.22 (m, 1H), 2.15-2.02 (m, 2H), 1.82-1.71 (m, 1H), 1.63-1.55 (m, 2H), 1.40-1.28 (m, 1H), 1.15 (d, J= 6.6 Hz, 4H). LCMS (ES, m/z): 512 [M+H]t Example 1.7: HPLC Conditions [0178] In any of the foregoing embodiments, the percentage purity recited may be determined by HPLC. In some embodiments the percentage purity is determined using the following HPLC
method:
Parameters Values Mobile Phase A 10 mM Ammonium Acetate Mobile Phase B Acetonitrile Parameters Values Column Waters XSelect Phenyl-Hexyl, 3.5 p.m, 4.6x150 mm Column Temperature 35 C
0 min 10%B
min 30%B
min 45%B
LC Gradient 21 min 80%B
22 min 80%B
22.1 min 10%B
Runtime 25 min LC Flow Rate 1 mL/min UV Wavelength 238 nm Ionization Mode Electrospray Ionization- Positive Mode Injection Volume 8 jut Example 2: HTRF biochemical assay for CBP and BRD4 activity [0179] The ability of compounds of Formula (I) to selectively inhibit CBP was determined using the following HTRF biochemical assay for CBP and BRD4 activity. The assay was performed in a final volume of 6 i.iL in assay buffer containing 50 mM Hepes pH 7.5, 0.5 mM
GSH, 0.01%
BGG, 0.005% BSA and 0.01% Triton X-100. Nanoliter quantities of 10-point, 3-fold serial dilution in DMSO were pre-dispensed into 1536 assay plates with a top concentration of 33 i.tM
and half log dilutions. 3 i.iL of 2x Protein and 3 i.iL of 2 x Peptide Ligand were added to assay plates (pre-stamped with compound). Plates were incubated for varying times up to 4 hours at room temperature prior to measuring the TR-FRET signal. IC50 values are shown in Figure 1. As set forth in Figure 1, an IC50 value of greater than or equal to 0.001 11M and less than or equal to 0.01 11M is marked "++++"; a value greater than 0.01 11M and less than or equal to 0.1 11M is marked "+++"; a value greater than 0.111M and less than or equal to 111M is marked "++"; and a value greater than 111M and less than 100011M is marked "+".
Example 3: Compounds 1 and Compound 1 demonstrated in vitro activity against CBP
[0180] The potency and selectivity of CBP/P300 inhibitor compounds including Compound 1 were determined in biochemical time resolved fluorescence assays using GST
fusions of the bromodomains of CBP and BRD4. Briefly, CBP inhibitors were pre-dispensed into 1536 assay plates for a final test concentration of 33 i.tM to 1.7 nM. Plates and incubated for 4h. Data were reported as percent inhibition compared with control wells. IC5o values were determined by curve fitting of the standard 4 parameter logistic fitting algorithm. In these conditions, Compound 1 was determined to be a potent inhibitor of CBP with an IC5o < 2 nM (N=16). In a similar assay, BRD4 potency was determined and Compound 1 showed an IC5o of <500 nM (N=15), indicating >200-fold selectivity.
[0181] Selectivity of Compound 1 was evaluated in screening assays for kinase inhibition and BRD binding. Compound 1 showed no to low binding affinity for the human kinases and disease-relevant mutant variants evaluated in a KINOMEscanTm screen. A panel of 10 BRD
representing the various branches or the bromodomain tree were tested using an AlphaScreen.
Of the 10 bromodomains surveyed, Compound 1 was inactive against 8. Compound 1 IC5o values for bromodomains of CREBBP and BRD4 (tandem BD1/BD2) were 0.1 and >10 [tM, respectively, confirming the high selectivity of Compound 1 for CBP.
[0182] The ability of Compound 1 and Compound 1 'to selectively inhibit CBP
was determined using the biochemical assay of Example 2 for CBP and BRD4 activity. Results are shown in Table 1 below:
Table 1 Compound CBP (IC5o) Selectivity Ratio of BRD4 (IC5o)/CBP (IC5o) 1 <10 nM >240 l' <20 nM >76 2 <10 nM 530 4 <10 nM 742 [0183] Both Compound 1 and Compound 1' potently inhibited (e.g., IC5o < 100 nM) CBP in the HTRF biochemical assay of Example 2, while Compound 1 was about 3.5-times more selective for CBP inhibition compared to BRD4 using this assay.

Example 4: Compound 1 demonstrates modulation of H3K27Ac in vitro in breast cancer cells [0184] AR-positive breast cancer cells were exposed to increasing concentrations of Compound 1 for 24 hours. Lysates were prepared in E-PAGE loading buffer (Invitrogen) and analyzed by western blot with antibodies diluted 1:1000 for anti-H3K27Ac, 1:2000 for anti-total H3 and 1:10,000 for anti-13 actin. The blots were scanned and analyzed on a LI-COR Odyssey image analyzer.
H3K27Ac levels were normalized to 13-Actin.
[0185] Compound 1 induced a concentration-dependent reduction of H3K27Ac, a mark specific to CBP/P300, in an AR positive breast cancer cell line. (Figure 3). Fifty percent reduction was seen between 0.03 and 0.1 M with maximal reduction of approximately 60% seen at 0.3 M.
Example 5: Compound 1, 2 and 4 reduce AR target gene TMPRSS2 and ER target gene XBP1 in AR-positive breast cancer cells [0186] AR-positive breast cancer cells were exposed to increasing concentrations of compounds for 24 hours. RNA was extracted and gene expression measured using Taqman assays for TMPRSS2 and XBP1.
[0187] All compounds reduced the mRNA expression of TMPRSS2 and XBP1 in an AR-positive breast cancer cell line.
[0188] As set forth in Table 2, below, an IC50 value less than 100 nM is marked "+++"; a value greater than 100 nM and less than 500 nM is marked "++"; and a value greater than 500 nM is marked "+".
Table 2 Compound 4 Compound 1 Compound 2 TMPRSS2 ++ +++ +
XBP1 ++ +++ +++
Example 6: Compounds 1, 2 and 4 have antiproliferative activity against AR+
breast cancer cell lines in vitro [0189] Breast cell lines were cultured according to the distributor's recommendations. The following day, cells were exposed to compounds continuously for 10 days. Cell viability was assessed using a CellTiter-Glo assay (Promega) at the end of the incubation period. The growth inhibitory effect was assessed by the concentration inhibiting growth by 50%
using a nonlinear regression equation and a variable slope (Graphpad Prism).
[0190] Compound 1, 2 and 4 inhibit proliferation of breast cancer cell lines after 10 days continuous exposure to the drug. The cell lines with high expression of AR
mRNA are more sensitive than those with low expression.
[0191] As set forth in Table 3, below, an IC50 value of less than 0.2 11M is marked "++++"; a value greater than 0.211M and less than 0.5 11M is marked "+++"; a value greater than 0.5 11M and less than 111M is marked "++"; and a value greater than 111M is marked "+".
Table 3 AR
Compound 4 Compound 1 Compound 2 MDA-MB-453 High ++++ ++++ +++
CAMA1 High ++++ ++++ ++++
HCC1187 High +++ +++ +++
HCC1500 High + ++ ++
BT549 Low + + +
CAL148 Low + + +
MFM223 Low + + +
MDA-MB-231 Low + + +
Example 7: Compound 1 demonstrated in vivo efficacy in AR-positive human derived breast cancer xenografts (TNBC) [0192] The antitumor activity of Compound 1 was tested in an AR-positive cell line derived xenograft model of AR+ triple negative breast cancer (Robinson et al., "Androgen receptor driven transcription in molecular apocrine breast cancer is mediated by FoxAl," The EMBO Journal (2011) 30, 3019-3027 (2011), incorporated herein by reference in its entirety). Briefly, AR-positive breast cancer tumor cells (1x107) were implanted subcutaneously in the flank of 6-8-week-old NOD SCID mice. Mice were randomized, and treatment started when mean tumor size reached 160 mm3 (8 mice per cohort). 50 mg/kg Compound 1 was administered orally daily for the duration of the experiment. Tumor volume (TV) was measured twice weekly by caliper and the tumor volume (mm3) calculated as follows: TV=a x b x b/2, where "a" and "b" are long and short diameters of a tumor, respectively.
[0193] Results are shown in Figure 4. At the end of the treatment period, Compound 1 treatment produced a tumor growth inhibition (TGI) of 104% (p<0.001) compared with vehicle control (TGI,[1-(TreatedTVfinal-TreatedTVinitial)/(VehicleTVfinal-VehicleTVinitial)]*100 where "TVfinal" and "TVinitial" are the mean tumor volumes on the final day and initial day of dosing).
The average body weight loss was 3.7%.
Example 8: Compound 2 modulates the level of AR and variants at the protein level.
[0194] AR-v7+ Prostate cancer cells were exposed to Compound 2 for 24 hours at which time lysates were prepared and the impact of the compound on the protein level of AR was assessed by western blot. The results are shown in Figure 5. Treatment of prostate cancer cells with Compound 2 led to the reduction of both full length and variant forms of the AR
including AR-v7.
Example 9: Compounds 1, 2 and 4 demonstrate modulation of AR target genes and KLK3 as well as MYC in an AR-v7+ prostate cancer cell line [0195] AR-v7+ prostate cancer cells were exposed to compounds 1, 2, and 4 for 24h. RNA was extracted using Qiacube RNAeasy Mini (Qiagen). For all genes tested, the qPCR reactions were carried out in triplicates with 250 ng RNA per reaction and Taqman primer-probes. GAPDH was used as a housekeeping gene.
[0196] In the conditions tested, all 3 compounds reduced AR target genes TMPRSS2 and KLK3, as well as MYC in a concentration-dependent manner in the AR-v7+ prostate cancer cells.
[0197] As set forth in Table 4, below, an IC50 value of less than 10 nM is marked "++++"; a value greater than 10 nM and less than 50 nM is marked "+++"; a value greater than 50 nM and less than 100 nM is marked "++"; and a value greater than 100 nM is marked "+".

Table 4 Target gene Compound 4 Compound 1 Compound 2 KLK3 +++ +++ ++
TMPRS S2 +++ ++++ ++
MYC N/A + +
Example 10: Compounds 1,2 and 4 demonstrate antiproliferative activity in prostate cancer cell lines [0198] Prostate cancer cells, androgen-dependent (LnCaP and VCaP) and androgen-independent (22Rv1), were plated and incubated overnight. The following day, cells were exposed to Compounds 1, 2, 4, and enzalutamide (final top concentration 10 p.M, half-log dilutions) continuously. Cell viability was assessed using a CellTiter-Glo assay (Promega) after 10 days drug exposure. The growth inhibitory effect was assessed by the concentration inhibiting growth by 50% using a nonlinear regression equation and a variable slope (Graphpad Prism).
[0199] Enzalutamide was active against androgen-dependent cell lines LnCaP and VCaP but was inactive in AR negative (PC3 and DU145) and the AR-v7 expressing 22Rv 1 cell line.
Conversely, all compounds had a potent and concentration-dependent growth inhibitory effect in all AR+ cell lines including 22Rv1 cells. All compounds were inactive in AR¨
cell lines.
[0200] As set forth in Table 5, below, an IC50 value of less than 0.511M is marked "+++"; a value greater than 0.5 11M and less than 111M is marked "++"; and a value greater than 1 11M is marked 4,1_,,.
Table 5 Cell Line AR status Compound 4 Compound 1 Compound 2 Enzalutamide LNCaP Androgen- +++ + +
dependent VCAP Androgen- +++ ++ + +++
dependent Cell Line AR status Compound 4 Compound 1 Compound 2 Enzalutamide 22Rv1 Androgen- ++ ++ ++ +
independent AR-v7+
PC-3 AR negative + + +
DU 145 AR negative + + +
Example 11: Compound 4 demonstrates antitumor activity in a patient-derived xenograft model of prostate cancer resistant to enzalutamide [0201] Male NOG mice (6-8 weeks of age) were implanted with prostate PDX tumor fragments.
When tumors reached an average tumor volume of 100-300 mm3 animals were randomized into different cohort groups and dosing initiated on the same day (Day 0). Compound 4 was formulated in (0.5 CMC/0.5% Tween 80) pH8 and administered as a solution. Animals were weighed twice weekly. Tumors were measured twice weekly. The maximum tumor volume of control animals was 1500 mm3.
[0202] Compound 4 was administered by oral gavage at a dose and schedule of 40 mg/kg/dose daily Mon-Thu repeated weekly or 80 mg/kg/dose Mo and Thu (twice weekly) repeated weekly.
[0203] Treatment with Compound 4 at 40 mg/kg/dose daily Mon-Thu repeated weekly or 80 mg/kg/dose Mo and Thu (twice weekly) repeated weekly resulted in a strong antitumor responss (Figure 6) with tumor growth inhibition values of 84% and 82%, respectively.
Enzalutamide had modest activity (TGI = 21%).
Example 12: Pharmaceutical Composition comprising Compound 1 and Compound 1' [0204] A pharmaceutical composition can comprise one or more compounds of formula (I), as provided herein, including Compound 1 and/or Compound F.
[0205] In one example, an active pharmaceutical ingredient (API) can comprise about 90% or more of Compound 1 and up to about 10% (preferably up to about 5%, most preferably up to about 2.5% including about 1.5%) of Compound 1'.

[0206] Oral dosage forms comprising Compound 1 can be prepared as a drug-in-capsule (DiC), encapsulated simple dry-blend granulation, and lipid-based solution in hard shell capsule. The capsules can contain pharmaceutically acceptable excipients, and encapsulated capsules can be packaged in high-density polyethylene induction sealed bottles.
[0207] Further embodiments of the disclosure are set out in the following numbered clauses:
1. A method of treating a patient diagnosed with an enzalutamide-resistant form of AR+
cancer, the method comprising administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition comprising Compound 1, or a pharmaceutically acceptable salt thereof:

OH
N-1.
2. A method of treating a patient diagnosed with an enzalutamide-resistant form of AR+
cancer, the method comprising administering to a patient in need thereof a therapeutically effective amount of a pharmaceutical composition comprising (1R,3R)-3-[(7S)-2-[(R)-(5-fluoro-2-methoxyphenyl)(hydroxy)methyl] -6-(methoxycarbony1)-7 -methyl-3H,6H,7H,8H,9Himidazo [4,5-f] quinolin-3 -yl] cyclohexane-l-carboxylic acid or a pharmaceutically acceptable salt thereof.
3. The method of clause 1 or clause 2, wherein Compound 1 is administered to the patient in an oral unit dosage form.

4. The method of any one of clauses 1-3, wherein the pharmaceutical composition further comprises Compound l' or a pharmaceutically acceptable salt thereof:

..10H
N-N

(1).
5. The method of clause 4, wherein the pharmaceutical composition comprises about 95% or more (by HPLC) of Compound 1 and up to 5% by weight of Compound 1'.
6. The method of one of clauses 1-5, wherein the AR+ cancer is an AR+
castrate resistant prostate cancer (CRPC).
7. The method of any one of clauses 1-5, wherein the AR+ cancer is an AR+
breast cancer.
8. The method of clause 7, wherein the patient is diagnosed with TNBC.
9. The method of any one of clauses 1-8, wherein the patient is diagnosed with a cancer harboring the AR-v7 spliceform of the Androgen Receptor.

Claims (36)

WO 2020/190792 PCT/US2020/022823We claim:

1. A method of treating breast cancer in a patient comprising administering to the patient in need thereof a therapeutically effective amount of a pharmaceutical composition comprising a compound of formula (I):
(12_ )n \ /
Ri N¨

N----9._ N OH

(D(D (I) or a pharmaceutically acceptable salt thereof, wherein:
R1 is H or ¨OH;
each R2 is independently selected from Ci-C6 alkyl, halogen, ¨CN, and ¨0R3, wherein the alkyl is optionally substituted with one or more halogen;
each R3 is independently C1-C6 alkyl, wherein the alkyl is optionally substituted with one or more halogen; and n is 0, 1, 2, or 3.

2. The method of claim 1, wherein the breast cancer is triple negative breast cancer.

3. The method of claim 1, wherein the breast cancer is AR+.

4. The method of claim 1, wherein the compound is selected from:
F

lik F 4.0 0>¨F
OH OH OH
No.9.... 0 No.,9.
N N N

, and , , o 19 ,s, OH
N....0 N

1 .

5. The method of claim 4, wherein the breast cancer is triple negative breast cancer.

6. The method of claim 4, wherein the breast cancer is AR+.

7. The method of claim 1, wherein the compound is:

OH
N¨

N

I .

8. The method of claim 1, wherein the compound is:

OH
N-

9. The method of claim 1, wherein the compound is:
F F0)¨F
OH
N¨

No.

10. The method of claim 1, wherein the compound is:
,( ,s, OH

11. The method of claim 1, wherein the breast cancer is AR+, and wherein the compound is selected from:

OH OH OH

, and N

12. The method of claim 11, wherein the breast cancer is triple negative breast cancer.

13. A method of treating prostate cancer in a patient comprising administering to the patient in need thereof a therapeutically effective amount of a pharmaceutical composition comprising a compound of formula (I):
(12_)n N ¨
N
OH

0 0 (I) or a pharmaceutically acceptable salt thereof, wherein:
R1 is H or ¨OH;
each R2 is independently selected from Ci-C6 alkyl, halogen, ¨CN, and ¨0R3, wherein the alkyl is optionally substituted with one or more halogen;

each R3 is independently Ci-C6 alkyl, wherein the alkyl is optionally substituted with one or more halogen; and n is 0, 1, 2, or 3.

14. The method of claim 13, wherein the prostate cancer is AR+.

15. The method of claim 13, wherein the prostate cancer is AR-v7+.

16. The method of claim 13, wherein the patient is diagnosed with castration-resistant prostate cancer or metastatic castration-sensitive prostate cancer.

17. The method of claim 13, wherein the compound is selected from:
F
F ilfr 01 . F ilfr 0)¨F
OH
OH OH
N¨

N¨ N¨

N N N

and = OH
N...,C) N

I .

18. The method of claim 17, wherein the patient is diagnosed with disease progression following treatment with enzalutamide.

19. The method of claim 13, wherein the compound is selected from:
F 40 01 *
OH
OH o N¨

N¨ $----N,..9.. 9_ N...,0 ; OH
N N

I I , and I
, .

20. The method of claim 19, wherein the prostate cancer is AR+.

21. The method of claim 20, wherein the prostate cancer is AR-v7+.

22. The method of claim 19, wherein the patient is diagnosed with castration-resistant prostate cancer or metastatic castration-sensitive prostate cancer.

23. A method of treating an Androgen Receptor-expressing cancer in a patient comprising administering to the patient in need thereof a therapeutically effective amount of a pharmaceutical composition comprising (1R,3R)-3-[(7S)-2-[(R)-(5-fluoro-2-methoxyphenyl)(hydroxy)methyl[-6-(methoxycarbony1)-7-methy1-3H,6H,7H,8H,9H-imidazo[4,5-fiquinolin-3-yl[cyclohexane-1-carboxylic acid, or a pharmaceutically acceptable salt thereof.

24. The method of claim 23, wherein the Androgen Receptor-expressing cancer is AR+ breast cancer.

25. The method of claim 24, wherein the AR+ breast cancer is triple negative breast cancer.

26. The method of claim 24, wherein the AR+ breast cancer is Her2- breast cancer.

27. The method of claim 24, wherein the AR+ breast cancer is selected from ER- breast cancer, PR- breast cancer, and ER-/PR- breast cancer.

28. The method of claim 23, wherein the Androgen Receptor-expressing cancer is AR+
prostate cancer.

29. The method of claim 28, wherein the AR+ prostate cancer is AR-v7+.

30. The method of claim 23, wherein the Androgen Receptor-expressing cancer is castration-resistant prostate cancer or metastatic castration-sensitive prostate cancer.

31. The method of claim 29, wherein the patient is diagnosed with disease progression following treatment with enzalutamide.

32. A method of treating an Androgen Receptor-expressing cancer in a patient comprising administering to the patient in need thereof a therapeutically effective amount of a pharmaceutical composition comprising:
Ii NOH

or a pharmaceutically acceptable salt thereof.

33. The method of claim 32, wherein the Androgen Receptor-expressing cancer is AR+ breast cancer.

34. The method of claim 22, wherein the AR+ breast cancer is triple negative breast cancer.

35. The method of claim 32, wherein the Androgen Receptor-expressing cancer is AR+
prostate cancer.

36. The method of claim 35, wherein the AR+ prostate cancer is AR-v7+.