Inhibition of ring AMP-Response Element Binding Protein (CREB)

Abstract

In one aspect, the disclosure relates to inhibition of ring AMP-response element binding protein (CREB). The present disclosure relates to solid and salt forms of inhibitors of CBP / p300 family bromodomains consisting of salts and crystalline forms of formula (I). Compounds may be used to treat diseases or conditions associated with inhibition of the CBP / p300 family bromodomains. For example, the disclosure relates to compounds and compositions for inhibiting the CBP / p300 family bromodomains, methods of treating diseases or conditions associated with inhibition of the CBP / p300 family bromodomains (e.g., certain forms of cancer), and methods of synthesizing these compounds.

Description

[0001] This application is a divisional application of the Chinese invention patent application (application date: March 13, 2020; application number: 202080028527.9 (International application number: PCT / US2020 / 022783); invention name: Inhibition of cyclic AMP-responsive element binding protein (CREB)).

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims the benefit of U.S. Provisional Patent Application No. 62 / 819,482, filed on March 15, 2019, which is hereby incorporated by reference in its entirety. Technical Field

[0004] The present disclosure relates to solid and salt forms of compounds and methods of inhibiting p300 (also known as EP300 and KAT3B) binding protein and / or cyclic AMP-responsive element binding protein (CREB) (CBP, also known as KAT3A) (a cellular paralog of p300) of adenovirus E1A protein. The compounds are useful for treating certain forms of cancer. Background Art

[0005] CBP / p300 is a lysine acetyltransferase that catalyzes the attachment of acetyl groups to lysine side chains of histones and other protein substrates. p300 (also known as EP300 and KAT3B) is a protein with multiple domains that bind to a variety of proteins including many DNA-binding transcription factors. Cyclic AMP-responsive element binding protein (CREB) binding protein (CBP, also known as KAT3A) is a cellular paralog of p300. p300 and CBP share extensive sequence identity and functional similarities, and are usually referred to as CBP / p300. The acetylation of histones and other proteins catalyzed by CBP / p300 is key to gene activation. Elevated p300 expression and activity have been observed in advanced human cancers such as prostate cancer and human primary breast cancer specimens. Chemical inhibition of CBP / p300 with intrinsic acetyltransferase enzymatic activity is more feasible than blocking transcription factors with small molecules, because the discovery of chemical inhibitors of transcription factors has proven to be extremely challenging.

[0006] Therefore, there is a need for new and highly effective compounds for inhibiting CBP / p300 that may be used as therapeutics for treating certain related forms of cancer. Summary of the invention

[0007] In one aspect, disclosed are compounds of formula (I):

[0008]

[0009] and non-amorphous solid forms of its salts.

[0010] In another aspect, disclosed are compounds having the stereochemistry of formula (II):

[0011]

[0012] and solid forms of their salts.

[0013] In some embodiments, the salt is an acid addition salt selected from hydrochloric acid, p-toluenesulfonic acid, benzenesulfonic acid, and sulfuric acid. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] This application includes drawings for better understanding of the principles of the present disclosure:

[0015] Figure 1 .A is a table of parameters used for X-ray powder diffraction analysis of the solid forms disclosed herein.

[0016] Figure 1 .B is a table of parameters used for thermogravimetric and differential scanning calorimetric analysis of the solid forms disclosed herein.

[0017] Figure 1 .C is a table of parameters used for dynamic vapor sorption analysis of the solid forms disclosed herein.

[0018] Figure 1 .D is a table of parameters used for HPLC analysis of amorphous free base compounds disclosed herein.

[0019] Figure 2 is a table of X-ray powder diffraction peaks having a relative intensity greater than 5% for the hydrochloric acid addition salt forms A, B and C disclosed herein.

[0020] Figure 3 TABLE 1 is a table of X-ray powder diffraction peaks having a relative intensity greater than 5% for the p-toluenesulfonic acid addition salt forms Form A and Form B disclosed herein.

[0021] Figure 4 is a table of X-ray powder diffraction peaks having a relative intensity greater than 5% for the benzenesulfonic acid addition salt form Type A disclosed herein.

[0022] Figure 5 is a table of X-ray powder diffraction peaks having a relative intensity greater than 5% for the sulfuric acid addition salt form Type A disclosed herein.

[0023] Figure 6 is a table of X-ray powder diffraction peaks having a relative intensity greater than 5% for the Form A crystalline free form solid form disclosed herein.

[0024] Figure 7is a table evaluating the kinetic solubility of solid forms of the hydrochloric acid addition salt form Form A, the p-toluenesulfonic acid addition salt form Form B, and the crystalline free form Form A disclosed herein. DETAILED DESCRIPTION

[0025] The present disclosure relates to salts and solid forms of compounds and compositions capable of modulating the activity of CBP / p300 family bromodomains. The present disclosure is characterized in that a method of treating, preventing or ameliorating a disease or condition in which CBP / p300 bromodomains play a role is provided by administering a therapeutically effective amount of a compound of formula (I), (II) or group A or a pharmaceutically acceptable salt thereof to a patient in need thereof. The methods disclosed herein can be used to treat a variety of CBP / p300 bromodomain-dependent diseases and conditions by inhibiting the activity of CBP / p300 bromodomains. Inhibition of CBP / p300 bromodomains provides a novel method for treating diseases (including but not limited to cancer).

[0026] Salts and crystalline solid forms of pharmaceutical compounds can confer several distinct advantages over amorphous or non-solid forms, including: 1) increased solubility, dissolution rate, and bioavailability of poorly soluble compounds, 2) reduced solubility for use in sustained release formulations, reduced Ostwald ripening, or to achieve taste masking of particularly soluble compounds, 3) improved physical properties such as melting temperature, hygroscopicity, and mechanical properties, 4) improved chemical stability and compatibility with pharmaceutical excipients, and 5) improved compound purity, chiral resolution of different stereoisomers, and filterability.

[0027] In certain embodiments, novel CBP inhibitor compounds are provided. Unless otherwise indicated, as used herein, a "CBP inhibitor compound" refers to a compound that has a detectable CBP IC when tested according to the HTRF biochemical assay protocol of Example 3. 50 Compounds with values ​​of 1 micromolar or less.

[0028] Unless otherwise indicated herein, the present disclosure provides all isomeric forms of a given compound, including mixtures thereof (e.g., S, R, and racemic orientations at each chiral center). If a compound contains a double bond, the substituents may be in the E or Z configuration. If a compound contains a disubstituted cycloalkyl, the cycloalkyl substituents may have a cis or trans configuration. All tautomeric forms are also intended to be included.

[0029] Unless otherwise stated, the compounds of formula (I), formula (II) and Group A may exist in their tautomeric forms. All such tautomeric forms are considered herein as part of the present disclosure.

[0030] Unless otherwise indicated, formula (I), formula (II) and group A compounds may contain one or more stereocenters and therefore exist in different stereoisomeric forms. Unless otherwise indicated, all stereoisomeric forms of formula (I), formula (II) and group A compounds and mixtures thereof (including racemic mixtures) are intended to be part of this disclosure. In addition, the present disclosure includes all geometric and positional isomers. For example, if formula (I), formula (II) or group A compounds contain double bonds or fused rings, cis and trans forms and mixtures are included in the scope of the present disclosure. Each compound disclosed herein includes all enantiomers that conform to the general structure of the compound. These compounds may be in racemic or enantiomerically pure form, or any other form of stereochemistry. The assay results may reflect data collected for racemic form, enantiomerically pure form or any other form of stereochemistry.

[0031] Diastereomeric mixtures can be separated into their respective diastereomers based on their physicochemical differences, for example, by chromatography and / or fractional crystallization, by methods well known to those skilled in the art. By reacting with suitable optically active compounds (for example, chiral auxiliary agents, such as chiral alcohols or Mosher acyl chlorides), enantiomeric mixtures are converted into diastereomeric mixtures, diastereomers are separated and single diastereomers are converted (for example, hydrolyzed) into corresponding pure enantiomers, enantiomers can be separated. In addition, some formula (I), formula (II) or group A compounds can be atropisomers (for example, substituted biaryl compounds), and are considered to be a part of the present disclosure. Enantiomers can also be separated by using chiral HPLC columns.

[0032] Formula (I), formula (II) or group A compounds can form acid addition salts, which can be pharmaceutically acceptable salts. The present disclosure also includes pharmaceutical compositions, which include one or more compounds described herein or pharmaceutically acceptable salts thereof, and pharmaceutically acceptable carriers. In some embodiments, the pharmaceutical compositions reported herein can be provided in unit dosage forms (e.g., capsules, tablets, etc.). In some embodiments, the pharmaceutical compositions reported herein can be provided in oral dosage forms. In some embodiments, the oral dosage forms of formula (I), formula (II) or group A compounds can be capsules. In some embodiments, the oral dosage forms of formula (I), formula (II) or group A compounds are tablets. In some embodiments, the oral dosage forms include one or more fillers, anthelmintics, lubricants, glidants, antiadhesives and / or antistatic agents. In some embodiments, oral dosage forms are prepared by dry mixing. In some embodiments, the oral dosage forms are tablets and are prepared by dry granulation.

[0033] The CBP inhibitor compounds of the present disclosure may be administered at therapeutically effective levels.

[0034] Compounds of the Disclosure

[0035] The present disclosure relates to a compound capable of modulating a CBP / p300 family bromodomain or a solid form of a pharmaceutically acceptable salt or isomer thereof, which can be used to treat diseases and disorders associated with the modulation of a CBP / p300 family bromodomain. The present disclosure also relates to a compound capable of inhibiting a CBP / p300 family bromodomain or a solid form of a pharmaceutically acceptable salt or isomer thereof.

[0036] In one aspect, the present disclosure relates to compounds of formula (I)

[0037]

[0038] and solid forms of enantiomers, hydrates, solvates, isomers and tautomers thereof, and acid addition salts thereof.

[0039] In some embodiments, the present disclosure relates to one or more selected from Group A:

[0040]

[0041]

[0042] and solid and salt forms of enantiomers, hydrates, solvates and tautomers thereof, as well as acid addition salts thereof.

[0043] In a preferred form, the present disclosure relates to solid and salt forms of formula (II):

[0044]

[0045] and their enantiomers, hydrates, solvates and tautomers, as well as acid addition salts thereof.

[0046] The above acid addition salts can be derived from the addition of hydrochloric acid (HCl), p-toluenesulfonic acid, benzenesulfonic acid and sulfuric acid (H2SO4). In the absence of acid addition salts, the compounds are mentioned in free base form. Acid addition salts and free base forms can be crystalline.

[0047] In some embodiments, the free base form may not be amorphous. The amorphous free base form is obtained by the synthesis provided in Example 1. This chemical synthesis has produced a compound of formula (II) with a relatively high purity. Except that compound A6 exists in a quantifiable amount, there are only trace amounts of stereoisomerism pollutants of group A in the final synthetic product. An amorphous free base form with a purity exceeding 99% (as measured by the HPLC analysis outlined in the following examples) was produced.

[0048] Regarding the solid form of hydrochloric acid addition salt, the applicant has found three crystalline forms of the compound of formula (I) (hereinafter referred to as Type A, Type B and Type C), two crystalline forms of p-toluenesulfonic acid addition salt (hereinafter referred to as Type A and Type B), one crystalline form of benzenesulfonic acid addition salt (hereinafter referred to as Type A), one form of sulfuric acid addition salt (hereinafter referred to as Type A) and a free base crystalline form (hereinafter referred to as free form Type A).

[0049] HCl addition salt form

[0050] Type A

[0051] The crystalline form A of the hydrochloric acid addition salt is characterized in that when Cu K α An X-ray powder diffraction pattern (XRPD) having at least three approximate peak positions (°2θ±0.2) selected from the group consisting of: 7.27, 8.98, 10.60, 15.60 and 23.93 when measured by radiometric measurement, when the XRPD is collected from about 3 degrees to about 40 degrees 2θ. Figure 2 A list of peaks in the X-ray powder diffraction pattern of the hydrochloric acid addition salt Form A containing relative intensities greater than or equal to 5%.

[0052] The HCl Form A salt is also characterized by an endothermic peak having an onset temperature of about 230° C. as measured by differential scanning calorimetry (DSC). The HCl Form A salt is also characterized by a weight loss of about 1.1% at temperatures up to 170° C. as measured by thermogravimetric analysis. The HCl Form A salt is also characterized by hygroscopicity as demonstrated by a water absorption of 6.3% at up to 80% relative humidity as measured by a dynamic vapor sorption isotherm plot. In another embodiment, the hydrochloric acid addition salt Form A is characterized by the kinetic solubility data shown in Example 2 below. The hydrochloric acid addition salt Form A is stable at temperatures up to 40° C. and up to 75% relative humidity for at least two weeks.

[0053] The crystalline form A of the hydrochloric acid addition salt is an anhydrate (anhydrous).

[0054] Form A of the hydrochloric acid addition salt can be prepared by dissolving the free base form in an organic solvent, adding hydrochloric acid, heating the resulting solution, and then cooling it as provided in Example 1.b.

[0055] Type B

[0056] The crystalline form B of the hydrochloric acid addition salt is characterized in that when Cu K αAn X-ray powder diffraction pattern (XRPD) having at least three approximate peak positions (°2θ±0.2) selected from the group consisting of: 10.23, 18.72, 23.03, 24.77 and 28.03 when measured by radiometric measurement, wherein the XRPD is collected from about 3 degrees to about 40 degrees 2θ. Figure 2 A list of peaks in the X-ray powder diffraction pattern of the hydrochloric acid addition salt Form B containing a relative intensity greater than or equal to 5%.

[0057] The HCl Form B salt is also characterized by endothermic peaks with onset temperatures of about 139° C. and 232° C. as measured by DSC and an exothermic peak at about 104° C. The HCl Form B salt is also characterized by a weight loss of about 20% at temperatures up to 200° C. as measured by thermogravimetric analysis.

[0058] In some embodiments, Form B of the hydrochloric acid addition salt is a hydrate or a solvate.

[0059] Form B of the hydrochloric acid addition salt can be prepared by dissolving Form A of the hydrochloric acid addition salt in a mixture of organic solvents as provided in Example 1.c, and then heating the resulting solution followed by cooling and slow evaporation at room temperature.

[0060] Type C

[0061] The hydrochloric acid addition salt of Form C is characterized in that when Cu K α An X-ray powder diffraction pattern (XRPD) having at least three approximate peak positions (°2θ±0.2) selected from the group consisting of: 7.05, 19.84, 21.09, 24.98 and 31.44 when measured by radiometric measurement, wherein the XRPD is collected from about 3 degrees to about 40 degrees 2θ. Figure 2 A list of peaks in the X-ray powder diffraction pattern of the hydrochloric acid addition salt Form B containing a relative intensity greater than or equal to 5%.

[0062] The HCl Form C salt is also characterized by endothermic peaks with onset temperatures of about 83° C., 143° C., and 179° C. as measured by DSC, and an exothermic peak at about 230° C. The HCl Form C salt is also characterized by a weight loss of about 9.4% at temperatures up to 180° C. as measured by thermogravimetric analysis.

[0063] In some embodiments, Form C of the hydrochloric acid addition salt is a hydrate or a solvate.

[0064] Form B of the hydrochloric acid addition salt can be prepared by dissolving Form A of the hydrochloric acid addition salt in a mixture of organic solvents and then heating the resulting solution followed by cooling and slow evaporation at room temperature as provided in Example 1.d.

[0065] p-Toluenesulfonic acid addition salt

[0066] Type A

[0067] The crystalline form A of the p-toluenesulfonic acid addition salt is characterized in that when Cu K α An X-ray powder diffraction pattern (XRPD) having at least three approximate peak positions (°2θ±0.2) selected from the group consisting of: 6.59, 13.20, 14.46, 18.00 and 21.74 when measured by radiometric measurement, wherein the XRPD is collected from about 3 degrees to about 40 degrees 2θ. Figure 3 A list of peaks in the X-ray powder diffraction pattern of p-toluenesulfonic acid addition salt Form A containing relative intensities greater than or equal to 5%.

[0068] p-Toluenesulfonic Acid Addition Salt Form A is also characterized by an endothermic peak having an onset temperature of about 74°C and 200°C as measured by DSC. p-Toluenesulfonic Acid Addition Salt Form A is also characterized by a weight loss of about 1.0% at temperatures up to 180°C as measured by thermogravimetric analysis.

[0069] Form A p-toluenesulfonic acid addition salt can be prepared by dissolving the free base form in an organic solvent, adding p-toluenesulfonic acid, heating the resulting solution, and then cooling as provided in Example 1.e.

[0070] Type B

[0071] The crystalline form B of the p-toluenesulfonic acid addition salt is characterized in that when Cu K α An X-ray powder diffraction pattern (XRPD) having at least three approximate peak positions (° 2θ ± 0.2) selected from the group consisting of: 7.10, 9.15, 15.08, 16.19, 17.25, 18.31 and 21.13 when measured by radiometric measurement, when the XRPD is collected from about 3 degrees to about 40 degrees 2θ. Figure 3 A list of peaks in the X-ray powder diffraction pattern of p-toluenesulfonic acid addition salt Form B containing relative intensities greater than or equal to 5%.

[0072] The p-toluenesulfonic acid addition salt Form B is also characterized by an endothermic peak having an onset temperature of about 225°C as measured by DSC. The p-toluenesulfonic acid addition salt Form B is also characterized by a weight loss of about 1.3% at temperatures up to 180°C as measured by thermogravimetric analysis. The p-toluenesulfonic acid addition salt Form B is also characterized by hygroscopicity as demonstrated by a water absorption of 2.1% at a relative humidity of up to 80% as measured by a dynamic vapor sorption isotherm plot. The p-toluenesulfonic acid addition salt Form B is characterized by the kinetic solubility data shown in Example 2 below. The p-toluenesulfonic acid addition salt Form B is stable at temperatures up to 40°C and relative humidity up to 75% for at least two weeks.

[0073] Form B p-toluenesulfonic acid addition salt can be prepared by dissolving the free base form in an organic solvent, adding p-toluenesulfonic acid, heating the resulting solution, and then cooling as provided in Example 1.f.

[0074] The crystalline form B of the p-toluenesulfonic acid addition salt is an anhydrate (anhydrous).

[0075] Benzenesulfonic acid addition salt type A

[0076] The crystalline form A of the benzenesulfonic acid addition salt is characterized in that when Cu K α An X-ray powder diffraction pattern (XRPD) having at least three approximate peak positions (°2θ±0.2) selected from the group consisting of: 5.84, 7.48, 9.45, 16.84, 18.90, 19.61, 20.70 and 25.14 when measured by radiation, wherein the XRPD is collected from about 3 degrees to about 40 degrees 2θ. Figure 4 A list of peaks in the X-ray powder diffraction pattern of benzenesulfonic acid addition salt Form A containing relative intensities greater than or equal to 5%.

[0077] Benzenesulfonic acid addition salt Form A is also characterized by an endothermic peak having an onset temperature of about 208° C. as measured by DSC. Benzenesulfonic acid addition salt Form A is also characterized by negligible weight loss at temperatures up to 210° C. as measured by thermogravimetric analysis.

[0078] Form A benzenesulfonic acid addition salt can be prepared by dissolving the free base form in an organic solvent, adding benzenesulfonic acid, heating the resulting solution, and then cooling as provided in Example 1.g.

[0079] Sulfate addition salt type A

[0080] The crystalline type A of the sulfuric acid addition salt is characterized in that when Cu K αAn X-ray powder diffraction pattern (XRPD) having at least three approximate peak positions (° 2θ ± 0.2) selected from the group consisting of: 8.24, 9.98, 13.58, 16.87, 18.78, 20.00 and 25.52 when measured by radiometric measurement, when the XRPD is collected from about 3 degrees to about 40 degrees 2θ. Figure 5 A list of peaks in the X-ray powder diffraction pattern of sulfuric acid addition salt Form A containing relative intensities greater than or equal to 5%.

[0081] Sulfate Addition Salt Form A is also characterized by endothermic peaks having onset temperatures of about 140° C. and 188° C. as measured by DSC. Sulfate Addition Salt Form A is also characterized by negligible weight loss at temperatures up to 200° C. as measured by thermogravimetric analysis.

[0082] Form A sulfuric acid addition salt can be prepared by dissolving the free base form in an organic solvent, adding sulfuric acid, heating the resulting solution, and then cooling as provided in Example 1.h.

[0083] Free base form A

[0084] The free base form of Form A is characterized in that when Cu K α An X-ray powder diffraction pattern (XRPD) having at least three approximate peak positions (°2θ±0.2) selected from the group consisting of: 11.11, 14.11, 18.21, 20.48 and 26.24 when measured by radiometric measurement, wherein the XRPD is collected from about 3 degrees to about 40 degrees 2θ. Figure 6 A list of peaks in the X-ray powder diffraction pattern of Form A free base having a relative intensity greater than or equal to 5%.

[0085] The free base form A is also characterized by an endothermic peak with an onset temperature of about 209° C. as measured by DSC. The free base form A is also characterized by negligible weight loss at temperatures up to 210° C. as measured by thermogravimetric analysis. The free base form A is characterized by the kinetic solubility data shown in Example 2 below.

[0086] Form A free base form can be prepared by dissolving the free base form in an organic solvent under reflux, followed by cooling the resulting solution, as provided in Example 1.i.

[0087] In some embodiments, the salts and crystalline forms used to prepare the pharmaceutical compositions are at least 95% pure as assessed by HPLC analysis.

[0088] Method for synthesizing the compound

[0089] The compounds of the present disclosure can be prepared by a variety of methods, including standard chemical methods. Suitable synthetic routes are described in the examples given below.

[0090] Compounds of the present disclosure, i.e. compounds of formula (I), (II) or group A or pharmaceutically acceptable salts thereof, can be prepared by methods as known in the field of organic synthesis, in part, by the synthetic schemes described in the examples. In the schemes described below, it is well known that, according to general principles or chemistry, protective groups of sensitive or reactive groups are used when necessary. Protective groups are operated according to standard methods of organic synthesis (TW Greene and PGM Wuts, "Protective Groups in Organic Synthesis", 3rd edition, Wiley, New York 1999). These groups are removed at a convenient stage of compound synthesis using methods that are obvious to those skilled in the art. The selection process and reaction conditions and their execution order should be consistent with the preparation of the group compounds of formula (I), (II) or group A.

[0091] Those skilled in the art will recognize that stereocenters are present in compounds of formula (I), (II) or group A. Therefore, the disclosure includes two possible stereoisomers (unless otherwise indicated and / or specified in the synthesis), not only including racemic compounds, but also including individual enantiomers and / or diastereomers. Unless otherwise indicated, when a compound needs to be used as a single enantiomer or diastereomer, it can be obtained by stereospecific synthesis or by the splitting of the final product or any convenient intermediate. The splitting of the final product, intermediate or starting material can be achieved by any suitable method known in the art. See, for example, " Stereochemistry of Organic Compounds " (Wiley-Interscience, 1994) by ELEiel, SH Wilen and LN Mander.

[0092] Methods of using the disclosed compounds

[0093] One aspect of the present disclosure relates to compounds of formula (I), (II) or Group A for use in medicine. Another aspect of the present disclosure relates to methods of modulating one or more CBP / p300 family bromodomains, comprising administering a therapeutically effective amount of a compound of formula (I), (II) or Group A to a patient in need thereof. Another aspect of the present disclosure relates to methods of inhibiting one or more CBP / p300 family bromodomains, comprising administering a therapeutically effective amount of a compound of formula (I), (II) or Group A to a patient in need thereof. In another aspect, the present disclosure relates to methods of inhibiting one or more CBP / p300 family bromodomains, comprising administering a therapeutically effective amount of a pharmaceutical composition comprising a compound of formula (I), (II) or Group A to a patient in need thereof.

[0094] CREB binding protein (CBP) inhibitor compounds can be used to develop pharmaceutical compositions suitable for treating certain related forms of cancer. CBP inhibitor compounds can be used to treat disease states that respond to the inhibition of CBP. CBP and EP300 (p300) are closely related multi-domain proteins that act as transcriptional coactivators. They carry acetyl lysine binding bromodomains that confer scaffolding or positioning functions to these proteins and have been shown to be suitable for designing small molecule inhibitors of their biological functions. These paralogs are highly homologous at the amino acid level and share many overlapping functions. They are histone acetyltransferases (HATs), which catalyze post-translational modifications of histones and non-histones. As HAT-carrying bromodomains, these proteins act as epigenetic readers and writers. The non-histone protein substrates of CBP / p300 are composed of many transcription factors, including nuclear hormone receptors, such as androgen receptor (AR). CBP / p300 acts as a coactivator of AR-signaling by acetylation AR, and the acetylation of the AR activates its transcriptional activity and promotes its protein stability. In addition, they acetylate histone H3 (Ac-H3K27) at lysine 27 to provide a docking site for the bromodomain, thereby providing a support for bridging the nuclear receptor to the basic transcription machinery complex (transcriptional machinery). The acetylation of histones leads to the generation of a transcription-permitting environment on chromatin. Therefore, the localization of CBP / p300 to the super-enhancer that AR relies on leads to an increase in the localized Ac-H3K27, which further increases the transcription of these loci.

[0095] Example

[0096] Material

[0097] Unless otherwise noted, all materials were obtained from commercial suppliers and 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 irradiation were performed on a Biotage Initiator EXP EU instrument.

[0098] Unless otherwise stated, mass-triggered HPLC purification and / or purity and low-resolution mass spectrometry data were measured using either: (1) a 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 low-resonance electrospray positive ionization mode (ESI) (column: Acquity UPLC BEH C18 1.7 μm 2.1×50 mm; gradient: 5-100% solvent B (95 / 5 / 0.09%: acetonitrile / water / formic acid) in solvent A (95 / 5 / 0.1%: 10 mM ammonium formate / acetonitrile / formic acid) for 2.2 minutes, then 100-5% solvent B in solvent A for 0.01 minutes, then hold at 5% solvent B in solvent A for 0.29 minutes), or (2) a Waters Acquity UPLC system (Waters Acquity UPLC with Sample Organizer and Waters Micromass ZQ mass spectrometer) with UV detection at 220 nm and low-resonance electrospray positive ionization mode (ESI) (column: Acquity UPLC BEH C18 1.7 μm 2.1×50 mm; gradient: 5-100% solvent B (95 / 5 / 0.09%: acetonitrile / water / formic acid) in solvent A (95 / 5 / 0.1%: 10 mM ammonium formate / acetonitrile / formic acid) for 2.2 minutes, then 100-5% solvent B in solvent A for 0.01 minutes, then hold at 5% solvent B in solvent A for 0.29 minutes). HT2790Alliance High Performance Liquid Chromatography (HPLC) system (Waters 996PDA and Waters ZQ Single Quad mass spectrometer) with UV detection at 220 nm and 254 nm and low resonance electrospray ionization (positive / negative) mode (ESI) (column: XBridge Phenyl or C18, 5 μm 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 minutes, then hold at 95% solvent B in solvent A for 1 minute (purity and low resolution MS only).

[0099] instrument

[0100] In the following examples, the Si zero background holder was used with a Panalytical X'Pert 3 Powder XRPD X-ray powder diffraction (XRPD) was performed. The 2θ position was calibrated relative to a Panalytical Si reference standard disk. Figure 1 The XRPD parameters used are listed in A.

[0101] Thermogravimetric analysis (TGA) data were collected using TA Instruments' TA Q500 and Q550. Differential scanning calorimetry (DSC) was performed using TA Instruments' TA Q2000. DSC was calibrated with an indium reference standard and TGA was calibrated with a nickel reference standard. The detailed parameters used are listed in Figure 1 In B.

[0102] Dynamic Vapor Sorption (DVS) data were collected using SMSDVS Intrinsic from Surface Measurement Systems. The parameters for the DVS tests are listed in Figure 1 C.

[0103] HPLC Assessment of Purity of Drug Compounds: Drug compound samples for analysis were prepared at a concentration of 0.2 mg / mL in a 70:30 mixture of water and acetonitrile. The samples were then analyzed on a Waters Alliance e2695 liquid chromatograph equipped with a Waters QDa mass spectrometer and a Waters 2998 photodiode array detector. The parameters of the chromatography method are disclosed at Figure 1 .D.

[0104] The definitions used in the following scheme and elsewhere in this paper are as follows:

[0105] Ac-H3K27 Histone H3 acetylated at lysine residue 27

[0106] AR androgen receptor

[0107] BSA bovine serum albumin

[0108] CBP Cyclic AMP response element binding protein (also known as KAT3A) DMSO Dimethyl sulfoxide

[0109] DSC Differential Scanning Calorimetry

[0110] DVS Dynamic Vapor Sorption

[0111] ES Electrospray ionization

[0112] EtOH

[0113] FRET Forster Resonance Energy Transfer

[0114] H Hour

[0115] H2SO4 sulfuric acid

[0116] HAT Histone acetyltransferase

[0117] HATU 2-(3H-[1,2,3]triazolo[4,5-b]pyridin-3-yl)-1,1,3,3-tetramethyl Isouronium hexafluorophosphate HCl

[0118] Hex

[0119] HPLC High Performance Liquid Chromatography

[0120] HTRF High-throughput time-resolved FRET IC 50 Half maximum inhibitory concentration

[0121] L Liter

[0122] LCMS Liquid chromatography / mass spectrometry

[0123] M Moore

[0124] mL milliliters

[0125] mmol millimole

[0126] mg milligram

[0127] MHz Megahertz

[0128] MS

[0129] m / z mass / charge ratio

[0130] NH4Cl Ammonium chloride

[0131] nm nanometer

[0132] NMR Nuclear Magnetic Resonance

[0133] p300 EP300 (also known as KAT3B)

[0134] ppm parts per million

[0135] TGA Thermogravimetric Analysis

[0136] UPLC Ultra-Performance Liquid Chromatography

[0137] UV

[0138] XRPD X-ray powder diffraction

[0139] ZnI2 Zinc iodide The term "organic solvent" is well known to those skilled in the art, but may include chemical solvents such as acetone, acetonitrile, benzene, chloroform, 1,4-dioxane, diethyl ether, dichloromethane, dimethylacetamide, N,N-dimethylformamide, dimethyl sulfoxide, ethanol, ethyl acetate, hexane, isopropanol, methanol, N-methylpyrrolidone, pyridine, tetrahydrofuran, toluene, and water, in addition to those not specifically named.

[0140] Example 1 - Preparation of Compounds, Solid Forms and Salts

[0141] Example 1.a-Preparation of amorphous free base + form

[0142] (1R,3R)-3-[(7S)-2-[(R)-(5-fluoro-2-methoxyphenyl)(hydroxy)methyl]-6-(methoxycarbonyl)-7-methyl-3H,6H,7H,8H,9H-imidazo[4,5-f]quinolin-3-yl]cyclohexane-1-carboxylic acid (1);

[0143] (1R,3R)-3-[(7S)-2-[(S)-(5-fluoro-2-methoxyphenyl)(hydroxy)methyl]-6-(methoxycarbonyl)-7-methyl-3H,6H,7H,8H,9H-imidazo[4,5-f]quinolin-3-yl]cyclohexane-1-carboxylic acid (1')

[0144] Synthesis of Intermediate 2-(5-Fluoro-2-methoxyphenyl)-2-hydroxyacetic acid

[0145]

[0146] Step 1. 2-(5-Fluoro-2-methoxyphenyl)-2-[(trimethylsilyl)oxy]acetonitrile

[0147] 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 at room temperature for 1 hour. The resulting mixture was concentrated under vacuum. The crude product was purified by silica gel chromatography (eluting with 1:1 ethyl acetate / petroleum ether) to give 2-(5-fluoro-2-methoxyphenyl)-2-[(trimethylsilyl)oxy]acetonitrile (2.0 g, 79%) as a white solid.

[0148] Step 2. 2-(5-Fluoro-2-methoxyphenyl)-2-hydroxyacetic acid

[0149] A solution of 2-(5-fluoro-2-methoxyphenyl)-2-[(trimethylsilyl)oxy]acetonitrile (1.50 g, 5.92 mmol) in hydrochloric acid (10 mL, 12 M). The resulting solution was stirred at 25 ° C for 1 hour and then at 70 ° C for 2 hours. 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% in 30 minutes); detector, UV 254 nm) to give 2-(5-fluoro-2-methoxyphenyl)-2-hydroxyacetic acid (1.10 g, 93%) as a white solid.

[0150] Step 3. 6-Fluoro-2-methyl-5-nitroquinoline

[0151] A solution of trifluoromethanesulfonic acid (82.0mL, 0.923mol) in HNO3 (19.6mL, 0.437mol) was stirred at 0°C for 20 minutes. 6-Fluoro-2-methylquinoline (50.0g, 0.310mol) in dichloromethane (300mL) was subsequently added at 0°C. The resulting mixture was stirred at room temperature (25°C) for 15 hours. The reaction mixture was diluted with water (300mL). The pH value of the solution was adjusted to 8 with sodium bicarbonate (saturated aqueous solution). The resulting solution was extracted with dichloromethane (3x 300mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (eluted with 1:4 ethyl acetate / petroleum ether) to give 6-fluoro-2-methyl-5-nitroquinoline (60.0g, 94%) as a light yellow solid. LCMS (ES, m / z): 207 [M+H] + .

[0152] Step 4. (2S)-6-Fluoro-2-methyl-5-nitro-1,2,3,4-tetrahydroquinoline

[0153] A solution of (S)-(-)-MeO-BIPHEP (1.03 g, 1.77 mmol) and chloro(1,5-cyclooctadiene)iridium(I) dimer (538 mg, 0.80 mmol) in toluene (100 mL) was stirred at room temperature (25° C.) under nitrogen atmosphere for 30 minutes. I2 (410 mg, 1.62 mmol) and 6-fluoro-2-methyl-5-nitroquinoline (33.0 g, 0.160 mol) in toluene (100 mL) were then added. The resulting mixture was stirred at room temperature (25° C.) under hydrogen (50 atmospheres) for 20 hours. The resulting mixture was concentrated under vacuum and purified by silica gel chromatography (eluted with 1:1 ethyl acetate / petroleum ether) to give a crude product (35.0 g). The crude product was dissolved in ethyl acetate (230 mL) and D-camphorsulfonic acid (36.9 g, 0.158 mol) was added. The resulting solution was stirred at 60 ° C for 1 hour and then cooled to room temperature. The solid was collected by filtration and rinsed with ethyl acetate (120 mL). The solid was 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 (3x 120 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give (2S)-6-fluoro-2-methyl-5-nitro-1,2,3,4-tetrahydroquinoline (25.5 g, 76%) as a red solid. LCMS (ES, m / z): 211 [M+H] +

[0154] Step 5. 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 chloroformate (18.7 mL, 0.242 mol) in dichloromethane (150 mL) was stirred at room temperature (25 ° C.) for 3 hours. The reactants were 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 give (2S)-6-fluoro-2-methyl-5-nitro-1,2,3,4-tetrahydroquinoline-1-carboxylic acid methyl ester (29.8 g, 92%) as a yellow solid. LCMS (ES, m / z): 269 [M+H] + .

[0156] Step 6. Methyl (2S)-6-[[(1R,3R)-3-(methoxycarbonyl)cyclohexyl]amino]-2-methyl-5-nitro-1,2,3,4-tetrahydroquinoline-1-carboxylate

[0157] A solution of (2S)-6-fluoro-2-methyl-5-nitro-1,2,3,4-tetrahydroquinoline-1-formic acid methyl ester (29.6 g, 0.110 mol), pyridine (29.6 mL, 0.368 mol), potassium carbonate (30.5 g, 0.220 mol), (1R, 3R)-3-aminocyclohexane-1-formic acid methyl ester (25.6 g, 162.84 mmol) in DMSO (270 mL) was stirred at 90 ° C for 15 hours and then cooled to room temperature. The reaction was quenched by adding water (200 mL) and extracted with ethyl acetate (3 x 300 mL). The combined organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The crude product was purified by silica gel chromatography (eluted with 1:1 ethyl acetate / petroleum ether) to give (2S)-6-[[(1R,3R)-3-(methoxycarbonyl)cyclohexyl]amino]-2-methyl-5-nitro-1,2,3,4-tetrahydroquinoline-1-carboxylic acid methyl ester (32 g, 72%) as a red oil. LCMS (ES, m / z): 406 [M+H] + .

[0158] Step 7. Methyl (2S)-5-amino-6-[[(1R,3R)-3-(methoxycarbonyl)cyclohexyl]amino]-2-methyl-1,2,3,4-tetrahydroquinoline-1-carboxylate

[0159] A solution of (2S)-6-[[(1R,3R)-3-(methoxycarbonyl)cyclohexyl]amino]-2-methyl-5-nitro-1,2,3,4-tetrahydroquinoline-1-carboxylic acid methyl ester (31.0 g, 76.46 mmol), NH4Cl (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 at 80°C for 1 hour and then cooled to room temperature. The solid was filtered off. The resulting solution was diluted with water (300 mL) and extracted with ethyl acetate (3x400 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give (2S)-5-amino-6-[[(1R,3R)-3-(methoxycarbonyl)cyclohexyl]amino]-2-methyl-1,2,3,4-tetrahydroquinoline-1-carboxylic acid methyl ester (27.5 g, 92%) as a dark green solid. LCMS (ES, m / z): 376 [M+H] + .

[0160] Step 8. Methyl (2S)-5-[2-(5-fluoro-2-methoxyphenyl)-2-hydroxyacetamido]-6-[[(1R,3R)-3-(methoxycarbonyl)cyclohexyl]amino]-2-methyl-1,2,3,4-tetrahydroquinoline-1-carboxylate

[0161] A solution of 2-(5-fluoro-2-methoxyphenyl)-2-hydroxyacetic acid (240 mg, 1.20 mmol), HATU (228 mg, 0.60 mmol), (2S)-5-amino-6-[[(1R,3R)-3-(methoxycarbonyl)cyclohexyl]amino]-2-methyl-1,2,3,4-tetrahydroquinoline-1-carboxylic acid methyl ester (150 mg, 0.40 mmol), DIEA (0.19 mL, 1.20 mmol) in N,N-dimethylformamide (10 mL) was stirred at 25 ° C for 1 hour. The resulting solution was diluted with H2O (10 mL). The resulting solution was extracted with ethyl acetate (3x15 mL), and the organic layers were 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 (eluted with 3:2 ethyl acetate / petroleum ether) to give (2S)-5-[2-(5-fluoro-2-methoxyphenyl)-2-hydroxyacetamido]-6-[[(1R,3R)-3-(methoxycarbonyl)cyclohexyl]amino]-2-methyl-1,2,3,4-tetrahydroquinoline-1-carboxylic acid methyl ester (180 mg, 81%) as a yellow solid. LCMS (ES, m / z): 558 [M+H] + .

[0162] Step 9. Methyl (7S)-2-[(5-fluoro-2-methoxyphenyl)(hydroxy)methyl]-3-[(1R,3R)-3-(methoxycarbonyl)cyclohexyl]-7-methyl-3H,6H,7H,8H,9H-imidazo[4,5-f]quinoline-6-carboxylate.

[0163] A solution of (2S)-5-[2-(5-fluoro-2-methoxyphenyl)-2-hydroxyacetamido]-6-[[(1R, 3R)-3-(methoxycarbonyl)cyclohexyl]amino]-2-methyl-1,2,3,4-tetrahydroquinoline-1-carboxylic acid methyl ester (180 mg, 0.32 mmol) in AcOH (8 mL) was stirred at 60 ° C overnight. The reaction mixture was cooled and concentrated under vacuum. The resulting crude product was purified by silica gel chromatography (eluted with 1:1 ethyl acetate / petroleum ether) to give (7S)-2-[(5-fluoro-2-methoxyphenyl)(hydroxy)methyl]-3-[(1R, 3R)-3-(methoxycarbonyl)cyclohexyl]-7-methyl-3H,6H,7H,8H,9H-imidazo[4,5-f]quinoline-6-carboxylic acid methyl ester (120 mg, 69%) as a yellow solid. LCMS (ES, m / z): 540 [M+H] + .

[0164] Step 10. (1R,3R)-3-[(7S)-2-[(R)-(5-fluoro-2-methoxyphenyl)(hydroxy)methyl]-6-(methoxycarbonyl)-7-methyl-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-(methoxycarbonyl)-7-methyl-3H,6H,7H,8H,9H-imidazo[4,5-f]quinolin-3-yl]cyclohexane-1-carboxylic acid (1')

[0165] A solution of methyl (7S)-2-[(5-fluoro-2-methoxyphenyl)(hydroxy)methyl]-3-[(1R,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 preparative HPLC (column, XBridge Prep C18OBD column, 19x150 mm, 5um; mobile phase, A: water (containing 10mmol / L NH4HCO3) and B: ACN (15.0% to 29.0% in 14 minutes); detector, UV 220 / 254nm). The product was separated by chiral preparative HPLC (column, CHIRALPAK IE, 2x25cm, 5um; mobile phase, A: Hex (containing 0.1% FA) and B: ethanol (maintaining 50.0% ethanol in 12 minutes); detector, UV 220 / 254nm). The product fractions were concentrated to give (1R,3R)-3-[(7S)-2-[(R)-(5-fluoro-2-methoxyphenyl)(hydroxy)methyl]-6-(methoxycarbonyl)-7-methyl-3H,6H,7H,8H,9H-imidazo[4,5-f]quinolin-3-yl]cyclohexane-1-carboxylic acid (23.6 mg, 20%) and (1R,3R)-3-[(7S)-2-[(S)-(5-fluoro-2-methoxyphenyl)(hydroxy)methyl]-6-(methoxycarbonyl)-7-methyl-3H,6H,7H,8H,9H-imidazo[4,5-f]quinolin-3-yl]cyclohexane-1-carboxylic acid (23.8 mg, 20%) as white solids. Stereoisomer purity was determined by HPLC: column: CHIRALPAK IE-3, column size: 0.46 x 5 cm; 3 μm; mobile phase: Hex (0.1% FA): EtOH = 50:50, flow rate: 1.0 ml / min. Compound purity was estimated to be greater than 99% as determined by HPLC analysis.

[0166] First eluting isomer (1): 1H-NMR(CD3OD,400MHz)δ(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) ,2.31-2.09(m,3H),1.83-1.58(m,3H),1.49-1.21(m,2H),1.16(d,J=6.8Hz,3H). LCMS (ES, m / z): 526 [M+H] + .

[0167] Second eluting isomer (1'): 1 H-NMR(CD3OD,400MHz)δ(ppm):7.69-7.44(m,2H),7.44-7.29(m,1H),7.12-6.99(m,1 H),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=6.4Hz,3H). LCMS(ES,m / z):526[M+H] + .

[0168] Example 1.b Preparation of Type A Hydrochloric Acid Addition Salt

[0169] 900 mg of (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)cyclohexane-1-carboxylic acid was dissolved in 16.5 mL of ethyl acetate and 37% hydrochloric acid was added in 3 equal portions (57 uL each) with vigorous stirring. After the first two aliquots, the solid precipitated and redissolved, but the solid was still present at the third addition. The slurry was stirred in a 50°C oil bath for 2 hours and then slowly cooled to ambient temperature overnight. The white slurry was cooled in an ice bath for 90 minutes and filtered / washed with cold ethyl acetate. The white solid was air-dried to 908 mg (94% yield). LCMS (ES, m / z): 526 [M+H] + The sample was further characterized by XRPD, DSC and TGA. The results showed that according to XRPD, the sample was crystalline, consistent with the Form A HCL salt.

[0170] Example 1. Preparation of c-hydrochloric acid addition salt: Type B

[0171] 20 mg of (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)cyclohexane-1-carboxylic acid, hydrochloric acid addition salt Form A was dissolved in 0.4 mL of a 1:1 mixture of dichloromethane and isopropanol. The solution was heated to 50°C and then cooled to 5°C at a rate of 0.1°C per minute and then stirred at 5°C overnight. The solution was then warmed to room temperature and a precipitate was observed after a portion of the solvent was slowly and unassisted evaporated. The precipitate was collected by centrifugation, dried under vacuum at 40°C, and then characterized by XRPD, DSC, and TGA. The results showed that the sample was crystalline by XRPD, consistent with the hydrochloric acid addition salt Form B.

[0172] Example 1. Preparation of d-hydrochloric acid addition salt: Form C

[0173] 20 mg of (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)cyclohexane-1-carboxylic acid, hydrochloric acid addition salt Form A was dissolved in 0.4 mL of a 9:1 mixture of 1,4-dioxane and water. The solution was heated to 50°C and then cooled to 5°C at a rate of 0.1°C per minute and then stirred at 5°C overnight. The solution was then warmed to room temperature and a precipitate was observed after a portion of the solvent was slowly and unassisted evaporated. The precipitate was collected by centrifugation, dried under vacuum at 40°C, and then characterized by XRPD, DSC, and TGA. The results showed that the sample was crystalline by XRPD, consistent with the hydrochloric acid addition salt Form C.

[0174] Example 1. Preparation of e-toluenesulfonic acid addition salt: Type A

[0175] 20 mg of (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)cyclohexane-1-carboxylic acid was dissolved in 0.4 mL of ethyl acetate and mixed with 1 equivalent of p-toluenesulfonic acid. The resulting solution was stirred at room temperature for 3 days. The resulting solid was collected by centrifugation, dried under vacuum at 40°C, and subsequently analyzed by XRPD, DSC, TGA and 1 The results showed that the sample was crystalline by XRPD, which was consistent with the p-toluenesulfonic acid addition salt type A.

[0176] Example 1. Preparation of fB-type p-toluenesulfonic acid addition salt

[0177] 300.6 mg of (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)cyclohexane-1-carboxylic acid was dissolved in 4.0 mL of acetonitrile in a 20 mL vial to form a suspension. 107.34 mg of p-toluenesulfonic acid was dissolved in 1.5 mL of acetonitrile and slowly added to the suspension. The suspension was then heated to 50°C and then cooled to 5°C at a rate of 0.1°C per minute. After stirring at 5°C overnight, the resulting precipitate was collected by centrifugation and dried under vacuum at 40°C to give 231.8 mg of a white powder. XRPD, DSC, TGA and 1 The solid was characterized by HNMR, and the results showed that the sample was crystalline by XRPD, consistent with Form B p-toluenesulfonic acid addition salt.

[0178] Example 1. Preparation of g-benzenesulfonic acid addition salt: Type A

[0179] 20 mg of (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)cyclohexane-1-carboxylic acid was dissolved in 0.4 mL of ethyl acetate and mixed with 1 equivalent of benzenesulfonic acid. The resulting solution was stirred at room temperature for 3 days. After no precipitation was observed at the three-day mark, the solution was heated to 50°C and slowly cooled to 5°C at a rate of 0.1°C per minute. After stirring the solution at 5°C overnight, a solid precipitate was observed. The resulting solid was collected by centrifugation, dried under vacuum at 40°C, and subsequently analyzed by XRPD, DSC, TGA, and 1 The results showed that the sample was crystalline by XRPD, which was consistent with the type A benzenesulfonic acid addition salt.

[0180] Example 1. Preparation of h-sulfuric acid addition salt: Type A

[0181] 20 mg of (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)cyclohexane-1-carboxylic acid was dissolved in 0.4 mL of ethyl acetate and mixed with 1 equivalent of sulfuric acid. The resulting solution was stirred at room temperature for 3 days. After no precipitation was observed at the three-day mark, the solution was heated to 50°C and slowly cooled to 5°C at a rate of 0.1°C per minute. After stirring the solution at 5°C overnight, solid precipitation was observed. The resulting solid was collected by centrifugation, dried at 40°C under vacuum, and subsequently characterized by XRPD, DSC, and TGA. The results showed that the sample was crystalline by XRPD, consistent with a Type A sulfuric acid addition salt.

[0182] Example 1. Preparation of Type iA Crystalline Free Form

[0183] (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)cyclohexane-1-carboxylic acid (5.00 g, 9.51 mmol) was completely dissolved in hot ethyl acetate (67 mL) at reflux. Turbidity appeared after a brief cooling of several minutes. The hot slurry was slowly cooled to ambient temperature and then stirred for 1 hour. The slurry was then cooled to 0°C and stirred in an ice bath for 2 hours before filtering and washing the solids with cold ethyl acetate. 3.70 g (74% recovery) of a white powder of increasing purity after drying was isolated. LCMS (ES, m / z): 526 [M+H] + The sample was further characterized by XRPD, DSC and TGA. These results indicated that the sample was crystalline by XRPD, consistent with Form A free form.

[0184] Example 2 - Kinetic Solubility Evaluation

[0185] The solubility of Form A HCl salt, Form B tosylate salt, and Form A free form in water, SGF, FaSSIF, and FeSSIF was measured at 37°C with a solid loading of approximately 10 mg / mL (calculated as free base). All solubility samples were kept rotating at 25 rpm at 37°C and samples were taken at 1 h, 4 h, and 24 h. The supernatant was extracted by centrifugation before filtration and used for solubility and pH measurements. The residual solid was collected for XRPD characterization. The results are summarized in Figure 7 middle.

[0186] Example 3 - HTRF biochemical assay of CBP and BRD4 activity

[0187] The ability of amorphous compound 1 to selectively inhibit CBP was determined using the following HTRF biochemical assay for CBP and BRD4 activity. Compound 2 was used as a reference compound:

[0188]

[0189] The assay was performed in a final volume of 6 μL in assay buffer containing 50 mM Hepes (pH 7.5, (0.5M Hepes, pH 7.5 solution; Teknova H1575)), 0.5 mM GSH, 0.01% BGG (0.22 μM filtered, Sigma, G7516-25G), 0.005% BSA (0.22 μM filtered, EMD Millipore Corporation, 126575) and 0.01% Triton X-100 (Sigma, T9284-10L). Nanoliter amounts of 10-point, 3-fold serial dilutions in DMSO were pre-distributed into 1536 assay plates (Corning, #3724BC) with final test concentrations ranging from 33 μM to 1.7 nM, highest to lowest doses, respectively. 3 μ L 2x protein and 3 μ L 2x peptide ligands are added to the assay plate (pre-stamped with compound). Before measuring the signal, the flat plate is incubated at room temperature for different times. TR-FRET (time-resolved fluorescence resonance energy transfer) is measured on a PHERAstar plate reader (BMG, equipped with HTRF optical module [337 / 520 / 490]) or Envision plate reader (PerkinElmer, equipped with TRF laser unit, TRF double mirror D400 / D505 and emission filter M520 and M495). Data are reported as percent inhibition compared to control wells according to the following equation: % inhibition = 1 - ((TR-FRET ratio - AveLow) / (AveHigh - AveLow)), where TR-FRET ratio = (fluorescence at 520 nm / fluorescence at 490 nm) * 10000), AveLow = average TR-FRET ratio of no enzyme control (n = 32), AveHigh = average TR-FRET ratio of DMSO control (n = 32). IC50 values ​​were determined by curve fitting using a standard 4-parameter logistic fit algorithm, which is included in the Activity Base software package: IDBS XE Designer Model 205. The data were fitted using the Levenburg Marquardt algorithm. For all assay formats, data are reported as percent inhibition compared to control wells according to the following equation: % Inhibition = 100*((FLU-AveLow) / (AveHigh-AveLow)), where FLU = measured fluorescence, AveLow = mean fluorescence of no enzyme control (n=32), AveHigh = mean fluorescence of DMSO control (n=32). IC50 values ​​were determined by curve fitting using a standard 4-parameter logistic fit algorithm, which is included in the Activity Base software package: IDBS XEDesigner Model205.The data were fitted using the Levenburg Marquardt algorithm. An IC of compound 1 for inhibition of CBP was observed. 50 The measured values ​​were between 0.001 μM and 0.01 μM, and the IC for inhibition of BRD4 50 The measured values ​​were between 1.0 μM and 1000 μM. Compound 2 was observed to inhibit CBP with an IC 50 The measured values ​​were between 0.001 μM and 0.01 μM, and the IC for inhibition of BRD4 50 The measured values ​​were between 1.0 μM and 1000 μM.

[0190] Example 4 - Compound 1 and Compound 1' demonstrate in vitro activity against CBP

[0191] Including compound 1 and compound 1':

[0192]

[0193] The potency and selectivity of CBP / P300 inhibitor compounds, including , were determined in a biochemical time-resolved fluorescence assay using GST fusions of the bromodomains of CBP and BRD4. Briefly, CBP inhibitors were pre-distributed into 1536 assay plates with final test concentrations ranging from 33 μM to 1.7 nM. Proteins and ligands were added to the compounds to a final concentration of 2.5 nM CBP or BRD4 (N-terminal GST-CREBBP (1081-1197), BRD4 tandem domains) and 25 nM tetraacetylated H3 peptide plates and incubated for 4 h. Data are reported as percent inhibition compared to control wells. IC 50 The values ​​were determined by curve fitting using a standard 4-parameter logistic fit algorithm. Under these conditions, compound 1 was identified as a potent inhibitor of CBP with an IC 50 <2 nM (N=16). In a similar assay, the potency of BRD4 was determined and compound 1 showed an IC of <500 nM 50 (N=15), indicating >200-fold selectivity.

[0194] The selectivity of compound 1 was evaluated in screening assays for kinase inhibition and BRD binding. TM Human kinases and disease-associated mutant variants evaluated in the screen showed no to low binding affinity. A small set of 10 BRDs representing different branches or bromodomain trees were tested using AlphaScreen. Compound 1 had no activity against 8 of the 10 bromodomains investigated. IC of Compound 1 against bromodomains of CREBBP and BRD4 (tandem BD1 / BD2) 50 The values ​​were 0.1 μM and >10 μM, respectively, confirming the high selectivity of compound 1 for CBP.

[0195] The ability of Compound 1 and Compound 1' to selectively inhibit CBP was determined using the biochemical assay for CBP and BRD4 activity of Example 3. The results are shown in the following table:

[0196] Compound <![CDATA[CBP(IC 50 )]]> <![CDATA[BRD4(IC 50 ) / CBP(IC 50 )'s selectivity ratio]]> 1 <10nM >266 1′ <20nM >76

[0197] In the HTRF biochemical assay of Example 3, both Compound 1 and Compound 1′ efficiently inhibited (e.g., IC 50 <100 nM) CBP, and using this assay, compound 1 is approximately 3.5-fold selective for CBP inhibition over BRD4 inhibition.

[0198] Other embodiments of the present disclosure are set forth in the following numbered clauses:

[0199] 1. A compound of formula (I):

[0200]

[0201] and non-amorphous solid forms of its salts.

[0202] 2. The solid form of clause 1, wherein the compound has the stereochemistry of formula (II):

[0203]

[0204] and its salts.

[0205] 3. The solid form of any one of clauses 1 and 2, wherein the salt is an acid addition salt selected from: hydrochloric acid, p-toluenesulfonic acid, benzenesulfonic acid and sulfuric acid.

[0206] 4. The solid form of clause 3, wherein the salt is a hydrochloric acid addition salt.

[0207] 5. The solid form of clause 4, wherein the hydrochloric acid addition salt form is Form A, characterized by an X-ray powder diffraction pattern comprising at least three peak positions (°2θ±0.2) when measured using Cu Kα radiation, the peak positions being selected from: 7.27, 8.98, 10.60, 15.60 and 23.93.

[0208] 6. The solid form of clause 4, wherein the hydrochloric acid addition salt form is Form B, characterized in that it comprises an X-ray powder diffraction pattern comprising at least three peak positions (°2θ±0.2) when measured using CuKα radiation, the peak positions being selected from the group consisting of: 10.23, 18.72, 23.03, 24.77 and 28.03.

[0209] 7. The solid form of clause 4, wherein the hydrochloric acid addition salt form is Form C, characterized by an X-ray powder diffraction pattern comprising at least three peak positions (°2θ±0.2) when measured using Cu Ka radiation, the peak positions being selected from: 7.05, 19.84, 21.09, 24.98 and 31.44.

[0210] 8. The solid form of clause 3, wherein the salt is a p-toluenesulfonic acid addition salt.

[0211] 9. The solid form of clause 8, wherein the p-toluenesulfonic acid addition salt form is Form A, characterized by an X-ray powder diffraction pattern comprising at least three peak positions (°2θ±0.2) when measured using Cu Kα radiation, the peak positions being selected from: 6.59, 13.20, 14.46, 18.00 and 21.74.

[0212] 10. The solid form of clause 8, wherein the p-toluenesulfonic acid addition salt form is Form B, characterized by an X-ray powder diffraction pattern comprising at least three peak positions (°2θ±0.2 degrees) when measured using Cu Kα radiation, the peak positions being selected from: 7.10, 9.15, 15.08, 16.19, 17.25, 18.31 and 21.13.

[0213] 11. The solid form of clause 3, wherein the salt is a benzenesulfonic acid addition salt.

[0214] 12. The solid form of clause 11, wherein the benzenesulfonic acid addition salt form is Form A, characterized by an X-ray powder diffraction pattern comprising at least three peak positions (°2θ±0.2) when measured using CuKα radiation, the peak positions being selected from: 5.84, 7.48, 9.45, 16.84, 18.90, 19.61, 20.70 and 25.14.

[0215] 13. The solid form of clause 3, wherein the salt is a sulfuric acid addition salt.

[0216] 14. The solid form of clause 13, wherein the sulfuric acid addition salt form is Form A, characterized by an X-ray powder diffraction pattern comprising at least three peak positions (°2θ±0.2) when measured using Cu Ka radiation, the peak positions being selected from the group consisting of: 8.24, 9.98, 13.58, 16.87, 18.78, 20.00 and 25.52.

[0217] 15. The solid form of any one of clauses 1 and 2, wherein the solid form is a Form A free form, characterized by an X-ray powder diffraction pattern comprising at least three peak positions (°2θ±0.2) when measured using Cu Ka radiation, the peak positions being selected from the group consisting of: 11.11, 14.11, 18.21, 20.48 and 26.24.

[0218] 16. A method for preparing a solid form of a compound of clause 1, comprising:

[0219] The free base, the amorphous compound of formula (I) is suspended in an acidic organic solvent or a mixture of organic solvents to form a solution; the solution is heated; and the solution is cooled.

[0220] 17. A method for preparing a solid form of a compound of clause 15, comprising:

[0221] suspending the free base, the amorphous compound of formula (I) in an organic solvent;

[0222] heating the solution; and

[0223] The solution was cooled, and then filtered and the precipitated solid matter was washed.

[0224] 18. A pharmaceutical composition comprising the solid form of any one of clauses 1 to 15 and one or more pharmaceutically acceptable carriers, adjuvants and vehicles.

[0225] 19. A method of inhibiting one or more CBP / p300 family bromodomains in a patient, comprising administering to a patient in need thereof an effective amount of the solid form of any one of clauses 1-15 or the pharmaceutical composition of clause 18.

[0226] 20. A method of treating, preventing, inhibiting or eliminating a disease or condition associated with the activity of one or more CBP / p300 family bromodomains in a patient, comprising: administering to a patient in need thereof a therapeutically effective amount of the solid form of any one of clauses 1-15 or the composition of clause 18.

[0227] 21. The solid form of any one of clauses 1 to 15 or the composition of clause 18 for use in the manufacture of a medicament for treating a disease associated with inhibition of one or more CBP / p300 family bromodomains.

[0228] 22. Use of the solid form of any of clauses 1-15 or the composition of clause 18 in treating a disease associated with inhibition of one or more CBP / p300 family bromodomains. Alternative embodiments of the present disclosure are set forth in the following numbered clauses:

[0229] 1. A solid form of a compound of formula (II):

[0230]

[0231] wherein the solid form is a hydrochloric acid addition salt, and wherein the hydrochloric acid addition salt is a Form A hydrochloric acid addition salt.

[0232] 2. The solid form of clause 1, wherein the solid form is characterized by an X-ray powder diffraction pattern comprising at least three peak positions (°2θ±0.2) when measured using Cu Ka radiation, the peak positions selected from the group consisting of: 7.27, 8.98, 10.60, 15.60 and 23.93.

[0233] 3. The solid form of any one of clauses 1 and 2, wherein the solid form is characterized by an endothermic peak having an onset temperature of about 230°C as measured by differential scanning calorimetry.

[0234] 4. The solid form of any one of clauses 1-3, wherein the solid form is characterized by a weight loss of about 1.1% at temperatures up to 170°C as measured by thermogravimetric analysis.

[0235] 5. The solid form of any one of clauses 1 to 4, wherein the solid form is an anhydrate.

[0236] 6. The solid form of any one of clauses 1 to 5, wherein the solid form is hygroscopic.

[0237] 7. The solid form of any one of clauses 1 to 6, wherein the solid form is stable at temperatures up to 40°C and at relative humidity up to 75% for at least two weeks.

[0238] 8. A method for preparing a solid form of compound (II):

[0239]

[0240] The method comprises:

[0241] dissolving the compound in an organic solvent or a mixture of organic solvents to form a solution; and

[0242] adding hydrochloric acid to the solution;

[0243] wherein the solid form is a hydrochloric acid addition salt, and wherein the hydrochloric acid addition salt is a Form A hydrochloric acid addition salt.

[0244] 9. The process of clause 8, wherein the organic solvent is ethyl acetate.

[0245] 10. The process of any one of clauses 8 and 9, wherein the concentration of the hydrochloric acid is about 37% w / v.

[0246] 11. The method of any one of clauses 8 to 10, wherein the method further comprises heating the solution.

[0247] 12. The method of any one of clauses 8 to 11, wherein the method further comprises cooling the solution.

[0248] 13. A pharmaceutical composition comprising the solid form of any one of clauses 1 to 7 and one or more pharmaceutically acceptable carriers, adjuvants or vehicles.

[0249] 14. A method of inhibiting one or more CBP / p300 family bromodomains in a patient, the method comprising administering to a patient in need thereof a therapeutically effective amount of the solid form of any one of clauses 1-7 or the pharmaceutical composition of clause 13.

[0250] 15. A method of treating, preventing, inhibiting or eliminating a disease or condition associated with the activity of one or more CBP / p300 family bromodomains in a patient, the method comprising administering to a patient in need thereof a therapeutically effective amount of the solid form of any one of clauses 1 to 7 or the pharmaceutical composition of clause 13.

[0251] The present disclosure also relates to the following embodiments:

[0252] 1. A solid form of a compound of formula (II):

[0253]

[0254] wherein the solid form is a hydrochloric acid addition salt, and wherein the hydrochloric acid addition salt is a Form A hydrochloric acid addition salt.

[0255] 2. The solid form of embodiment 1, wherein the solid form is characterized by an X-ray powder diffraction pattern comprising at least three peak positions (°2θ±0.2) when measured using Cu Ka radiation, wherein the peak positions are selected from the group consisting of: 7.27, 8.98, 10.60, 15.60 and 23.93.

[0256] 3. The solid form of embodiment 1, wherein the solid form is characterized by an endothermic peak with an onset temperature of about 230°C as measured by differential scanning calorimetry.

[0257] 4. The solid form of embodiment 1, wherein the solid form is characterized by a weight loss of about 1.1% at temperatures up to 170°C as measured by thermogravimetric analysis.

[0258] 5. The solid form of embodiment 1, wherein the solid form is an anhydrate.

[0259] 6. The solid form of embodiment 1, wherein the solid form is hygroscopic.

[0260] 7. The solid form of embodiment 1, wherein the solid form is stable at temperatures up to 40°C and relative humidity up to 75% for at least two weeks.

[0261] 8. A method for preparing a solid form of compound (II):

[0262]

[0263] The method comprises:

[0264] dissolving the compound in an organic solvent or a mixture of organic solvents to form a solution; and

[0265] adding hydrochloric acid to the solution;

[0266] wherein the solid form is a hydrochloric acid addition salt, and wherein the hydrochloric acid addition salt is a Form A hydrochloric acid addition salt.

[0267] 9. The method of embodiment 8, wherein the organic solvent is ethyl acetate.

[0268] 10. The method of embodiment 8, wherein the concentration of the hydrochloric acid is about 37% w / v.

[0269] 11. The method of embodiment 8, wherein the method further comprises heating the solution.

[0270] 12. The method of embodiment 11, wherein heating the solution comprises heating the solution to a temperature of about 50°C.

[0271] 13. The method of embodiment 11, further comprising cooling the solution.

[0272] 14. The method of embodiment 13, wherein cooling the solution comprises cooling the solution to a temperature of about 20°C to about 25°C.

[0273] 15. The method of any one of embodiments 8, wherein the method further comprises filtering the solid form.

[0274] 16. A pharmaceutical composition comprising the solid form of embodiment 1 and one or more pharmaceutically acceptable carriers, adjuvants or vehicles.

[0275] 17. A method of inhibiting one or more CBP / p300 family bromodomains in a patient, comprising administering to a patient in need thereof a therapeutically effective amount of a solid form of embodiment 1.

[0276] 18. A method of inhibiting one or more CBP / p300 family bromodomains in a patient, comprising administering to a patient in need thereof a therapeutically effective amount of the pharmaceutical composition of embodiment 16.

[0277] 19. A method of treating breast cancer in a patient, comprising administering to the patient in need thereof a therapeutically effective amount of the solid form of embodiment 1.

[0278] 20. A method of treating prostate cancer in a patient, comprising administering to the patient in need thereof a therapeutically effective amount of the pharmaceutical composition of embodiment 16.

Claims

1. A solid form of a hydrochloric acid addition salt of a compound of formula (II): Characterized by an X-ray powder diffraction pattern comprising at least three peak positions (°2θ±0.2) selected from the group consisting of: 10.23, 18.72, 23.03, 24.77 and 28.03 when measured using Cu Ka radiation.

2. The solid form of claim 1, wherein the solid form is characterized by an endothermic peak having an onset temperature of 139°C or 232°C as measured by differential scanning calorimetry.

3. The solid form of claim 1, wherein the solid form is characterized by a 20% weight loss at temperatures up to 200°C as measured by thermogravimetric analysis.

4. The solid form of claim 1, wherein the solid form is a hydrate or a solvate.

5. The solid form of claim 1, wherein the solid form is hygroscopic.

6. The solid form of any one of claims 1-5, wherein the solid form is stable at temperatures up to 40°C and at relative humidity up to 75% for at least two weeks.

7. A method for preparing a hydrochloric acid addition salt of compound (II): The method comprises: Dissolve the A-type hydrochloric acid addition salt of (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)cyclohexane-1-carboxylic acid in an organic solvent; as well as heating the solution; and cooling the solution; Wherein when the hydrochloric acid addition salt is measured using Cu Ka radiation, it comprises an X-ray powder diffraction pattern of at least three peak positions (°2θ±0.2), wherein the peak positions are selected from the group consisting of: 10.23, 18.72, 23.03, 24.77 and 28.

03.

8. The method of claim 7, wherein the organic solvent is a 1:1 mixture of dichloromethane and isopropanol.

9. The method of claim 7, wherein heating the solution comprises heating the solution to a temperature of 50°C.

10. The method of claim 7, wherein cooling the solution comprises cooling the solution to a temperature of 5°C.

11. The method of claim 7, further comprising warming the solution to room temperature.

12. The method of claim 7, further comprising evaporating the solvent and collecting by centrifugation.

13. A pharmaceutical composition comprising the solid form of any one of claims 1 to 6 and one or more pharmaceutically acceptable carriers, adjuvants or vehicles.

14. Use of the solid form of any one of claims 1 to 6 for the preparation of a medicament for inhibiting one or more CBP / p300 family bromodomains in a patient.

15. Use of the pharmaceutical composition of claim 13 in the preparation of a medicament for inhibiting one or more CBP / p300 family bromodomains in a patient.

16. Use of a solid form as claimed in any one of claims 1 to 6 for the preparation of a medicament for treating breast cancer in a patient.

17. Use of the pharmaceutical composition of claim 13 in the preparation of a medicament for treating breast cancer in a patient.

18. Use of a solid form as claimed in any one of claims 1 to 6 for the preparation of a medicament for treating prostate cancer in a patient.

19. Use of the pharmaceutical composition of claim 13 in the preparation of a medicament for treating prostate cancer in a patient.

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