阅读说明：本技术 抑制环amp-应答元件结合蛋白(creb) (Inhibiting cyclic AMP-response element binding protein (CREB) ) 是由 G.卢克 于 2020-03-13 设计创作，主要内容包括：本公开涉及由式(I)的盐和晶型构成的CBP/p300家族溴结构域的抑制剂的固体和盐形式。化合物可用于治疗与所述CBP/p300家族溴结构域的抑制相关联的疾病或病症。例如,本公开涉及用于抑制所述CBP/p300家族溴结构域的化合物和组合物、治疗与所述CBP/p300家族溴结构域抑制相关联的疾病或病症(例如,某些形式的癌症)的方法以及这些化合物的合成方法。(The present disclosure relates to solid and salt forms of inhibitors of the CBP/p300 family bromodomain comprised of salts and crystalline forms of formula (I). The compounds are useful for treating diseases or disorders associated with the inhibition of the bromodomain of the CBP/p300 family. For example, the present disclosure relates to compounds and compositions for inhibiting the CBP/p300 family bromodomain, methods of treating diseases or disorders associated with the CBP/p300 family bromodomain inhibition (e.g., certain forms of cancer), and methods of synthesis of these compounds.)

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

wherein said solid form is a hydrochloric acid addition salt, and wherein said hydrochloric acid addition salt is the hydrochloric acid addition salt form A.

2. The solid form of claim 1, wherein said solid form is characterized by an X-ray powder diffraction pattern comprising at least three peak positions (° 2 Θ ± 0.2) selected from the group consisting of: 7.27, 8.98, 10.60, 15.60 and 23.93.

3. The solid form of claim 1, wherein the solid form is characterized by an endothermic peak having an onset temperature of about 230 ℃ as measured by differential scanning calorimetry.

4. The solid form of claim 1, wherein the solid form is characterized by a weight loss of about 1.1% at temperatures up to 170 ℃ as measured by thermogravimetric analysis.

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

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

7. The solid form of claim 1, wherein the solid form is stable for at least two weeks at a temperature of up to 40 ℃ and a relative humidity of up to 75%.

8. A process for preparing a solid form of compound (II):

the method comprises the following steps:

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

adding hydrochloric acid to the solution;

wherein said solid form is a hydrochloric acid addition salt, and wherein said hydrochloric acid addition salt is the hydrochloric acid addition salt form A.

9. The method of claim 8, wherein the organic solvent is ethyl acetate.

10. The method of claim 8, wherein the hydrochloric acid is at a concentration of about 37% w/v.

11. The method of claim 8, wherein the method further comprises heating the solution.

12. The method of claim 11, wherein heating the solution comprises heating the solution to a temperature of about 50 ℃.

13. The method of claim 11, wherein the method further comprises cooling the solution.

14. The method of claim 13, wherein cooling the solution comprises cooling the solution to a temperature of about 20 ℃ to about 25 ℃.

15. The method of any one of claims 8, wherein the method further comprises filtering the solid form.

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

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 the solid form of claim 1.

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 claim 16.

19. A method of treating breast cancer in a patient comprising administering to a patient in need thereof a therapeutically effective amount of the solid form of claim 1.

20. A method of treating prostate cancer in a patient comprising administering to a patient in need thereof a therapeutically effective amount of the pharmaceutical composition of claim 16.

Technical Field

The present disclosure relates to solid and salt forms of compounds and methods of inhibiting the p300 (also known as EP300 and KAT3B) binding protein and/or cyclic AMP-response element binding protein (CREB) (CBP, also known as KAT3A) (a cellular paralog of p300) of the adenovirus E1A protein. The compounds are useful in the treatment of certain forms of cancer.

Background

CBP/p300 is a lysine acetyltransferase that catalyzes the attachment of acetyl groups to lysine side chains of histone 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-response element binding protein (CREB) binding protein (CBP, also known as KAT3A) is a cellular paralogue of p 300. p300 and CBP share extensive sequence identity and functional similarity, commonly referred to as CBP/p 300. Acetylation of histones and other proteins catalyzed by CBP/p300 is critical for gene activation. Elevated p300 expression and activity has been observed in specimens of advanced human cancers such as prostate cancer and human primary breast cancer. Chemical inhibition of CBP/p300 with intrinsic acetyltransferase enzymatic activity is more feasible than blocking transcription factors with small molecules, as the discovery of chemical inhibitors of transcription factors has proven to be extremely challenging.

Thus, there is a need for new and highly potent compounds for inhibiting CBP/p300, which are useful as therapeutics for the treatment of certain related forms of cancer.

Disclosure of Invention

In one aspect, compounds of formula (I):

and non-amorphous solid forms of salts thereof.

In another aspect, compounds having the stereochemistry of formula (II):

and solid forms of salts thereof.

In some embodiments, the salt is an acid addition salt selected from the group consisting of hydrochloric acid, p-toluenesulfonic acid, benzenesulfonic acid, and sulfuric acid.

Drawings

The present application contains the accompanying drawings to better understand the principles of the disclosure:

FIG. 1.A is a table of parameters for X-ray powder diffraction analysis of the solid forms disclosed herein.

FIG. 1.B is a table of parameters for thermogravimetric and differential scanning calorimetry analyses of the solid forms disclosed herein.

Fig. 1.C is a table of parameters for dynamic vapor sorption analysis of the solid form disclosed herein.

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

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

Figure 3 is a table of X-ray powder diffraction peaks for the p-toluenesulfonic acid addition salt forms a and B disclosed herein with relative intensities of greater than 5%.

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

Figure 5 is a table of X-ray powder diffraction peaks for the form a sulfuric acid addition salt form disclosed herein with a relative intensity of greater than 5%.

Figure 6 is a table of X-ray powder diffraction peaks for crystalline form a free form solid forms disclosed herein having a relative intensity of greater than 5%.

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

Detailed Description

The present disclosure relates to salts and solid forms of compounds and compositions capable of modulating the activity of the bromodomain of the CBP/p300 family. The disclosure features methods of treating, preventing, or ameliorating diseases or conditions in which CBP/p300 bromodomains play a role 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 of the present disclosure are useful for treating a variety of CBP/p300 bromodomain-dependent diseases and disorders by inhibiting the activity of the CBP/p300 bromodomain. Inhibition of the CBP/p300 bromodomain provides a novel approach to the treatment of diseases, including but not limited to cancer.

Salts and crystalline solid forms of a pharmaceutical compound may confer several distinct advantages over amorphous or non-solid forms, including: 1) increasing the solubility, dissolution rate and bioavailability of poorly soluble compounds, 2) decreasing solubility for sustained release formulations, reducing Ostwald ripening, or achieving taste masking of particularly soluble compounds, 3) improving physical properties such as melting temperature, hygroscopicity and mechanical properties, 4) improving chemical stability and compatibility with pharmaceutically acceptable excipients, and 5) increasing compound purity, chiral resolution of different stereoisomers, and filterability.

In certain embodiments, novel CBP inhibitor compounds are provided. As used herein, unless otherwise specified, "CBP inhibitor compound" means a detectable CBP IC when tested according to the HTRF biochemical assay protocol of example 3₅₀Compounds with a value of 1 micromolar or less.

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

Unless otherwise indicated, 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.

Unless otherwise indicated, the compounds of formula (I), formula (II) and group a may contain one or more stereocenters and thus exist in different stereoisomeric forms. Unless otherwise indicated, all stereoisomeric forms of compounds of formula (I), formula (II) and group a and mixtures thereof (including racemic mixtures) are intended to form part of the present disclosure. In addition, the present disclosure includes all geometric and positional isomers. For example, if a compound of formula (I), formula (II) or group a contains a double or fused ring, both the cis and trans forms as well as mixtures are included within 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 in any other form of stereochemistry. The assay results may reflect data collected for racemic forms, enantiomerically pure forms, or any other form of stereochemistry.

Mixtures of diastereomers may be separated into their respective diastereomers on the basis of their physicochemical differences by methods well known to those skilled in the art, for example, by chromatography and/or fractional crystallization. Enantiomers can be separated by converting the enantiomeric mixture into a diastereomeric mixture by reaction with a suitable optically active compound (e.g., a 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. In addition, some compounds of formula (I), formula (II) or group a may be atropisomers (e.g., substituted biaryl compounds) and are considered part of this disclosure. Enantiomers can also be separated by using a chiral HPLC column.

The compounds of formula (I), formula (II) or group a may form acid addition salts, which may be pharmaceutically acceptable salts. The present disclosure also includes pharmaceutical compositions comprising one or more compounds described herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical compositions reported herein may be provided in unit dosage forms (e.g., capsules, tablets, etc.). In some embodiments, the pharmaceutical composition as reported herein may be provided in an oral dosage form. In some embodiments, the oral dosage form of a compound of formula (I), formula (II), or group a may be a capsule. In some embodiments, the oral dosage form of the compound of formula (I), formula (II), or group a is a tablet. In some embodiments, the oral dosage form comprises one or more fillers, insect repellents, lubricants, glidants, anti-adherents, and/or antistatic agents. In some embodiments, the oral dosage form is prepared by dry blending. In some embodiments, the oral dosage form is a tablet and is prepared by dry granulation.

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

Compounds of the present disclosure

The present disclosure relates to solid forms of compounds, or pharmaceutically acceptable salts or isomers thereof, capable of modulating a CBP/p300 family bromodomain, which are useful for treating diseases and disorders associated with modulation of a CBP/p300 family bromodomain. The disclosure also relates to solid forms of compounds useful for inhibiting the bromodomain of the CBP/p300 family, or pharmaceutically acceptable salts or isomers thereof.

In one aspect, the disclosure relates to compounds of formula (I)

And enantiomers, hydrates, solvates, isomers and tautomers thereof, as well as acid addition salts of the aforementioned substances.

In some embodiments, the present disclosure relates to one or more selected from group a:

and enantiomers, hydrates, solvates and tautomers thereof as well as solid and salt forms of acid addition salts of the aforementioned substances.

In preferred forms, the present disclosure relates to solid and salt forms of formula (II):

and enantiomers, hydrates, solvates and tautomers thereof, as well as acid addition salts of the aforementioned.

The above acid addition salts can be derived from hydrochloric acid (HCl), p-toluenesulfonic acid, benzenesulfonic acid and sulfuric acid (H)₂SO₄) Addition of (1). In the absence of acid addition salts, the compounds are mentioned in the form of the free bases. The acid addition salts and the free base forms may be crystalline.

In some embodiments, the free base form may not be amorphous. The amorphous free base form was obtained by the synthesis provided in example 1. This chemical synthesis yields a compound of formula (II) with a relatively high purity. Only trace amounts of the stereoisomeric contaminants of group a were present in the final synthesis product, except that compound a6 was present in quantifiable amounts. Amorphous free base forms with a purity of over 99% (as determined by HPLC analysis outlined in the examples below) were produced.

With regard to the solid forms of hydrochloric acid addition salts, the applicant found three crystalline forms of the compound of formula (I) (hereinafter referred to as form a, form B and form C), two crystalline forms of p-toluenesulfonic acid addition salts (hereinafter referred to as form a and form B), one crystalline form of benzenesulfonic acid addition salt (hereinafter referred to as form a), one crystalline form of sulfuric acid addition salt (hereinafter referred to as form a) and the free base crystalline form (hereinafter referred to as free form a).

HCl addition salt forms

Type A

Form A of the hydrochloric acid addition salt is characterized in that when Cu K is used_αAn X-ray powder diffraction pattern (XRPD) having at least three approximate peak positions (° 2 θ ± 0.2) when measured radiometrically, said approximate peak positions selected from the group consisting of: 7.27, 8.98, 10.60, 15.60, and 23.93, when XRPD was collected from about 3 degrees to about 40 degrees 2 θ. FIG. 2X-ray powder diffraction diagram of the hydrochloric acid addition salt form A comprising a relative intensity greater than or equal to 5%List of peaks.

The HCl a salt is also characterized by an endothermic peak having an onset temperature of about 230 ℃ as measured by Differential Scanning Calorimetry (DSC). The HCl a-type salt is also characterized by a weight loss of about 1.1% at temperatures up to 170 ℃, as measured by thermogravimetric analysis. The HCl a salt is also characterized by hygroscopicity as evidenced by a water absorption of 6.3% at a relative humidity of up to 80% as measured by a dynamic vapor sorption isotherm diagram. 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 for at least two weeks at temperatures up to 40 ℃ and relative humidities up to 75%.

The crystalline form a of the hydrochloric acid addition salt is anhydrous (anhydrous).

Crystalline 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, as provided in example 1. b.

Type B

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

The HCl B salt is also characterized by endothermic peaks with onset temperatures of about 139 ℃ and 232 ℃ and exothermic peaks of about 104 ℃ as measured by DSC. The HCl B-type salt is also characterized by a weight loss of about 20% at temperatures up to 200 ℃ as measured by thermogravimetric analysis.

In some embodiments, the crystalline form B of the hydrochloric acid addition salt is a hydrate or solvate.

Form B of the hydrochloric acid addition salt can be prepared by dissolving the hydrochloric acid addition salt form a 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.

C type

Form C of the hydrochloric acid addition salt is characterized in that when Cu K is used_αAn X-ray powder diffraction pattern (XRPD) having at least three approximate peak positions (° 2 θ ± 0.2) when measured radioactively, said approximate peak positions selected from the group consisting of: 7.05, 19.84, 21.09, 24.98, and 31.44, when XRPD was collected from about 3 degrees to about 40 degrees 2 θ. Figure 2 contains a list of the peaks of the X-ray powder diffraction pattern of the hydrochloric acid addition salt form B with a relative intensity greater than or equal to 5%.

The HCl form C salt is also characterized by endothermic peaks with onset temperatures of about 83 ℃, 143 ℃, and 179 ℃ and exothermic peaks of about 230 ℃ as measured by DSC. The HCl type C salt is also characterized by a weight loss of about 9.4% at temperatures up to 180 ℃ as measured by thermogravimetric analysis.

In some embodiments, the crystalline form C of the hydrochloric acid addition salt is a hydrate or solvate.

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

P-toluenesulfonic acid addition salts

Type A

Form A of the p-toluenesulfonic acid addition salt is characterized when using Cu K_αAn X-ray powder diffraction pattern (XRPD) having at least three approximate peak positions (° 2 θ ± 0.2) when measured radioactively, said approximate peak positions selected from the group consisting of: 6.59, 13.20, 14.46, 18.00, and 21.74, when XRPD was collected from about 3 degrees to about 40 degrees 2 θ. Figure 3 contains a list of the peaks of the X-ray powder diffraction pattern of the addition salt of p-toluenesulfonic acid form a with a relative intensity greater than or equal to 5%.

The p-toluenesulfonic acid addition salt of form a is further characterized by endothermic peaks with onset temperatures of about 74 ℃ and 200 ℃ as measured by DSC. The p-toluenesulfonic acid addition salt form a is further characterized by a weight loss of about 1.0% at temperatures up to 180 ℃ as measured by thermogravimetric analysis.

The addition salt of form a p-toluenesulfonic acid 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.

Type B

Form B of the p-toluenesulfonic acid addition salt is characterized when using Cu K_αAn X-ray powder diffraction pattern (XRPD) having at least three approximate peak positions (° 2 θ ± 0.2) when measured radioactively, said approximate peak positions selected from the group consisting of: 7.10, 9.15, 15.08, 16.19, 17.25, 18.31, and 21.13, when XRPD is collected from about 3 degrees to about 40 degrees 2 θ. Figure 3 contains a list of the peaks of the X-ray powder diffraction pattern of the addition salt of p-toluenesulfonic acid form B with a relative intensity greater than or equal to 5%.

The p-toluenesulfonic acid addition salt form B is further characterized by an endothermic peak with an onset temperature of about 225 ℃ as measured by DSC. The p-toluenesulfonic acid addition salt form B is further characterized by a weight loss of about 1.3% at temperatures up to 180 ℃ as measured by thermogravimetric analysis. The crystalline form B p-toluenesulfonic acid addition salt is further characterized by hygroscopicity as evidenced by water absorption of 2.1% at relative humidity up to 80% as measured by dynamic vapor sorption isotherm diagram. 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 for at least two weeks at temperatures up to 40 ℃ and relative humidities up to 75%.

The p-toluenesulfonic acid addition salt of form B 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.

Form B of the p-toluenesulfonic acid addition salt is anhydrous (anhydrous).

Benzenesulfonic acid addition salts of form A

Form A of a benzenesulfonic acid addition salt is characterized by the use of Cu K_αAt least three approximate peak positions (° 2 θ) when measured radiatively0.2) and an X-ray powder diffraction pattern (XRPD), said approximate peak positions being selected from: 5.84, 7.48, 9.45, 16.84, 18.90, 19.61, 20.70, and 25.14, when the XRPD was collected from 2 θ of about 3 degrees to about 40 degrees. Figure 4 contains a list of peaks in the X-ray powder diffraction pattern of the benzenesulfonic acid addition salt form a at relative intensities greater than or equal to 5%.

The benzenesulfonic acid addition salt form a is also characterized by an endothermic peak having an onset temperature of about 208 ℃ as measured by DSC. The benzenesulfonic acid addition salt form a is also characterized by negligible weight loss at temperatures up to 210 ℃ as measured by thermogravimetric analysis.

The addition salt of form a benzenesulfonic acid 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.

Sulfuric acid addition salts of form A

The crystal form A of the sulfuric acid addition salt is characterized in that when Cu K is used_αAn X-ray powder diffraction pattern (XRPD) having at least three approximate peak positions (° 2 θ ± 0.2) when measured radioactively, said approximate peak positions selected from the group consisting of: 8.24, 9.98, 13.58, 16.87, 18.78, 20.00, and 25.52, when XRPD is collected from about 3 degrees to about 40 degrees 2 θ. Figure 5 contains a list of the peaks of the X-ray powder diffraction pattern of the sulfuric acid addition salt form a with a relative intensity greater than or equal to 5%.

The form a sulfuric acid addition salt is also characterized by endothermic peaks with onset temperatures of about 140 ℃ and 188 ℃ as measured by DSC. The sulfuric acid addition salt form a is also characterized by a negligible weight loss at temperatures up to 200 ℃ as measured by thermogravimetric analysis.

The sulfuric acid addition salt of form a 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.

Free base crystalline form A

The free base form of form A is characterized when Cu K is used_αHaving at least three approximate peak positions (° 2 θ ± 0.2) when measured radiometricallyAn X-ray powder diffraction pattern (XRPD), said approximate peak positions selected from: 11.11, 14.11, 18.21, 20.48, and 26.24, when XRPD is collected from about 3 degrees to about 40 degrees 2 θ. Figure 6 contains a list of peaks in the X-ray powder diffraction pattern of form a free base form with a relative intensity greater than or equal to 5%.

The form a free base form is also characterized by an endothermic peak having an onset temperature of about 209 ℃, as measured by DSC. The free base form a is also characterized by negligible weight loss at temperatures up to 210 ℃ as measured by thermogravimetric analysis. The form a free base form is characterized by the kinetic solubility data shown below in example 2.

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.

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

Methods for synthesizing said compounds

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.

The compounds of the present disclosure, i.e., compounds of formula (I), (II) or group a, or pharmaceutically acceptable salts thereof, may be prepared by methods as are known in the art of organic synthesis, set forth in part by the synthetic schemes described in the examples. In the schemes described below, it is well known that protective groups for sensitive or reactive groups are used as necessary according to general principles or chemistry. The protecting Groups were manipulated according to standard methods of Organic Synthesis (T.W.Greene and P.G.M.Wuts, "Protective Groups in Organic Synthesis", 3 rd edition, Wiley, New York 1999). These groups are removed at a convenient stage of the compound synthesis using methods that will be apparent to those skilled in the art. The selection process as well as the reaction conditions and their order of execution should be consistent with the preparation of a group of compounds of formula (I), (II) or group a.

One skilled in the art will recognize that stereocenters exist in compounds of formula (I), (II) or group a. Thus, the present disclosure includes both possible stereoisomers (unless otherwise indicated and/or specified in the synthesis), including not only racemic compounds, but also individual enantiomers and/or diastereomers. 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, intermediate or starting material may be achieved by any suitable method known in the art. See, e.g., "Stereochemistry of Organic Compounds" by E.L.Eliel, S.H.Wilen, and L.N.Mander (Wiley-lnterscience, 1994).

Methods of using the disclosed compounds

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

CREB Binding Protein (CBP) inhibitor compounds are useful in the development of pharmaceutical compositions useful in the treatment of certain related forms of cancer. CBP inhibitor compounds are useful for treating disease states that respond to inhibition of CBP. CBP and EP300(p300) are closely related multidomain proteins that function as transcriptional co-activators. They carry an acetyl lysine-binding bromodomain that confers a scaffold or localization function to these proteins and have been demonstrated to be useful for designing small molecule inhibitors of their biological function. 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 a bromodomain carrying HAT, these proteins function as epigenetic readers (readers) and writers (writers). The non-histone protein substrates of CBP/p300 are composed of a number of transcription factors, including nuclear hormone receptors, such as the Androgen Receptor (AR). CBP/p300 acts as a co-activator of AR-signaling by acetylating AR, which activates its transcriptional activity and promotes its protein stability. In addition, they acetylated histone H3(Ac-H3K27) at lysine 27 to provide a docking site for the bromodomain, thereby providing a scaffold for bridging nuclear receptors to the basic transcription machinery complex. Acetylation of histones results in the production of a transcriptional permissive environment on chromatin. Thus, the positioning of CBP/p300 to the AR-dependent super-enhancer resulted in an increase in the positioned Ac-H3K27, which further increased transcription at these loci.

Examples

Material

Unless otherwise indicated, all materials were obtained from commercial suppliers and used without further purification. The anhydrous solvent was obtained from Sigma-Aldrich (Milwaukee, Wis.), and used as received. All reactions involving air or moisture sensitive reagents were performed under nitrogen atmosphere and all reactions using microwave irradiation were performed on a Biotage Initiator EXP EU instrument.

Unless otherwise indicated, mass triggered HPLC purification and/or purity and low resolution mass spectral data were measured using either: (1) waters acquisition ultra-high performance liquid chromatography (UPLC) system (Waters acquisition UPLC with Sample Organizer (Sample Organizer) and Waters Micromass ZQ mass spectrometer), UV detection at 220nm and with low resonance electrospray positive ion mode (ESI) (column: acquisition UPLC BEH C181.7 μm 2.1X 50 mM; gradient: 5-100% solvent B in solvent A (95/5/0.1%: 10mM ammonium formate/acetonitrile/formic acid) (95/5/0.09%: acetonitrile/water/formic acid) for 2.2 minutes, then 100-5% solvent B in solvent A for 0.01 minutes, then 5% solvent B in solvent A for 0.29 minutes), or (2) Waters HT2790 Alliance High Performance Liquid Chromatography (HPLC) system (Waters 996 Siji and Waters ZQ mass spectrometer), UV detection was carried out at 220nm and 254nm and with a low resonance electrospray ionization (positive/negative) mode (ESI) (column: Xbridge Phenyl or C18,5 μm4.6X50 mm; gradient: 5-95% of solvent B (95% methanol/5% methanol, 0.1% formic acid) in solvent A (95% water/5% methanol, 0.1% formic acid) for 2.5 minutes, followed by 1 minute hold in 95% of solvent B in solvent A (purity only and low resolution MS).

Instrument for measuring the position of a moving object

In the following examples, Panalytical X' Pert was used on a Si zero background holder³Powder XRPD X-ray powder diffraction (XRPD) was performed. The 2 θ position was calibrated against a Panalytical Si reference standard disc. The XRPD parameters used are listed in fig. 1A.

Thermogravimetric analysis (TGA) data was collected using TA Q500 and Q550 from TA Instruments. Differential Scanning Calorimetry (DSC) was performed using TA Q2000 from TA Instruments. DSC was calibrated with indium reference standards and TGA was calibrated with nickel reference standards. The details of the parameters used are set forth in FIG. 1B.

Dynamic Vapor Sorption (DVS) data was collected using SMS DVS Intrasic from Surface Measurement Systems. The parameters of the DVS test are listed in fig. 1C.

HPLC evaluation of the purity of the pharmaceutical compound: a sample of the pharmaceutical compound was prepared for analysis in a 70:30 mixture of water and acetonitrile at a concentration of 0.2 mg/mL. 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 chromatography are disclosed in fig. 1. D.

The following schemes and definitions used elsewhere herein are as follows:

histone H3 with Ac-H3K27 acetylated at lysine residue 27

AR androgen receptor

BSA bovine serum albumin

CBP cyclic AMP response element binding protein (also known as KAT3A)

DMSO dimethyl sulfoxide

DSC differential scanning calorimetry

DVS dynamic vapor adsorption

ES electrospray ionization

EtOH ethanol

FRET Forster resonance energy transfer

H hours

H₂SO₄Sulfuric acid

HAT histone acetyltransferase

HATU 2- (3H- [1,2,3] triazolo [4,5-b ] pyridin-3-yl) -1,1,3, 3-tetramethylisouronium hexafluorophosphate

HCl hydrochloric acid

Hex Hexane

HPLC high performance liquid chromatography

HTRF high-throughput time-resolved FRET

IC₅₀Half maximal inhibitory concentration

L liter

LCMS liquid chromatography/Mass Spectrometry

M mol

mL of

mmol millimole

mg of

MHz megahertz

MS Mass Spectrometry

mass/charge ratio of m/z

NH₄Cl ammonium chloride

nm nanometer

NMR nuclear magnetic resonance

P300 EP300 (also known as KAT3B)

ppm parts per million

TGA thermogravimetric analysis

UPLC ultra-high performance liquid chromatography

UV ultraviolet ray

XRPD X-ray powder diffraction

ZnI₂Zinc iodide

The term "organic solvent" is readily 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-dimethylformamide, dimethylsulfoxide, ethanol, ethyl acetate, hexane, isopropanol, methanol, N-methylpyrrolidone, pyridine, tetrahydrofuran, toluene, water, in addition to those not specifically named.

EXAMPLE 1 preparation of Compounds, solid forms and salts

Example 1.a preparation of amorphous free base + form

(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')

Synthesis of intermediate 2- (5-fluoro-2-methoxyphenyl) -2-hydroxyacetic acid

Step 1.2- (5-fluoro-2-methoxyphenyl) -2- [ (trimethylsilyl) oxy ] acetonitrile

ZnI is reacted at room temperature₂A solution of (1.6mg,0.01mmol, 5-fluoro-2-methoxybenzaldehyde (1.54g,9.99mmol) in trimethylsilylcarbonitrile (1.5mL,11.25mmol) was stirred for 1h the resulting mixture was concentrated in vacuo the resulting crude was purified by silica gel chromatography (eluting with 1:1 ethyl acetate/petroleum ether) to give 2- (5-fluoro-2-methoxyphenyl) -2- [ (trimethylsilyl) oxy as a white solid]Acetonitrile (2.0g, 79%).

Step 2.2- (5-fluoro-2-methoxyphenyl) -2-hydroxyacetic acid

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

Step 3.6-fluoro-2-methyl-5-nitroquinoline

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

Step 4.(2S) -6-fluoro-2-methyl-5-nitro-1, 2,3, 4-tetrahydroquinoline

A solution of (S) - (-) -MeO-BIPHEP (1.03g,1.77mmol), chloro (1, 5-cyclooctadiene) iridium (I) dimer (538mg,0.80mmol) in toluene (100mL) was stirred at room temperature (25 ℃ C.) under a nitrogen atmosphere for 30 minutes. Followed by addition of I in toluene (100mL)₂(410mg,1.62mmol), 6-fluoro-2-methyl-5-nitroquinoline (33.0g,0.160 mol). The resulting mixture was stirred at room temperature (25 ℃) under hydrogen (50 atm) for 20 hours. The resulting mixture was concentrated in vacuo and purified by silica gel chromatography (eluting with 1:1 ethyl acetate/petroleum ether) to give the crude product (35.0 g). The crude product was dissolved in ethyl acetate (230mL) and D-camphorsulfonic acid (36.9g,0.158mol) was added. The resulting solution was stirred at 60 ℃ for 1 hour and then cooled to room temperature. The solid was collected by filtration and washed with ethyl acetate (120 mL). The solid was dissolved in water (50 mL). The pH of the solution was adjusted to 8 with sodium bicarbonate (saturated aqueous solution). The resulting solution was extracted with ethyl acetate (3 × 120 mL). The combined organic matterThe layers were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give (2S) -6-fluoro-2-methyl-5-nitro-1, 2,3, 4-tetrahydroquinoline (25.5g, 76%) as a red solid. LCMS (ES, M/z) 211[ M + H]⁺

Step 5.(2S) -6-fluoro-2-methyl-5-nitro-1, 2,3, 4-tetrahydroquinoline-1-carboxylic acid methyl ester

A solution of (2S) -6-fluoro-2-methyl-5-nitro-1, 2,3, 4-tetrahydroquinoline (25.3g,0.120mol), pyridine (39.0mL,0.484mol), methyl chloroformate (18.7mL,0.242mol) in dichloromethane (150mL) was stirred at room temperature (25 ℃ C.) for 3 hours. The reaction was washed with 1M hydrochloric acid (2 × 70 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give (2S) -6-fluoro-2-methyl-5-nitro-1, 2,3, 4-tetrahydroquinoline-1-carboxylic acid methyl ester (29.8g, 92%) as a yellow solid. LCMS (ES, M/z):269[ M + H]⁺。

Step 6.(2S) -6- [ [ (1R,3R) -3- (methoxycarbonyl) cyclohexyl ] amino ] -2-methyl-5-nitro-1, 2,3, 4-tetrahydroquinoline-1-carboxylic acid methyl ester

A solution of methyl (2S) -6-fluoro-2-methyl-5-nitro-1, 2,3, 4-tetrahydroquinoline-1-carboxylate (29.6g,0.110mol), pyridine (29.6mL,0.368mol), potassium carbonate (30.5g,0.220mol), (1R,3R) -3-aminocyclohexane-1-carboxylate (25.6g,162.84mmol) in DMSO (270mL) was stirred at 90 ℃ for 15 hours and then cooled to room temperature. The reaction was quenched by the addition of water (200mL) and extracted with ethyl acetate (3 × 300 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The resulting crude product was purified by silica gel chromatography (eluting with 1:1 ethyl acetate/petroleum ether) to give (2S) -6- [ [ (1R,3R) -3- (methoxycarbonyl) cyclohexyl ] as a red oil]Amino group]-methyl 2-methyl-5-nitro-1, 2,3, 4-tetrahydroquinoline-1-carboxylate (32g, 72%). LCMS (ES, M/z) 406[ M + H]⁺。

Step 7.(2S) -5-amino-6- [ [ (1R,3R) -3- (methoxycarbonyl) cyclohexyl ] amino ] -2-methyl-1, 2,3, 4-tetrahydroquinoline-1-carboxylic acid methyl ester

Reacting (2S) -6- [ [ (1R,3R) -3- (methoxycarbonyl) cyclohexyl]Amino group]-methyl 2-methyl-5-nitro-1, 2,3, 4-tetrahydroquinoline-1-carboxylate (31.0g,76.46mmol), NH₄Cl(24.3g,454.28mmol)、Fe(64.3g,1.15mol) in tetrahydrofuran (300mL), ethanol (300mL), water (100mL) was stirred at 80 ℃ for 1 hour, then cooled to room temperature. The solid was filtered off by filtration. The resulting solution was diluted with water (300mL) and extracted with ethyl acetate (3 × 400 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give (2S) -5-amino-6- [ [ (1R,3R) -3- (methoxycarbonyl) cyclohexyl ] as a dark green solid]Amino group]-methyl 2-methyl-1, 2,3, 4-tetrahydroquinoline-1-carboxylate (27.5g, 92%). LCMS (ES, M/z) 376[ M + H]⁺。

Step 8.(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

2- (5-fluoro-2-methoxyphenyl) -2-hydroxyacetic acid (240mg,1.20mmol), HATU (228mg,0.60mmol), (2S) -5-amino-6- [ [ (1R,3R) -3- (methoxycarbonyl) cyclohexyl]Amino group]A solution of methyl (150mg,0.40mmol) of (E) -2-methyl-1, 2,3, 4-tetrahydroquinoline-1-carboxylate (0.19mL,1.20mmol) and DIEA in N, N-dimethylformamide (10mL) was stirred at 25 ℃ for 1 hour. Subjecting the resulting solution to H₂Dilution with O (10 mL). The resulting solution was extracted with ethyl acetate (3 × 15 mL) and the organic layers were combined. The resulting mixture was washed with brine (2 × 20 mL). The mixture was dried over anhydrous sodium sulfate and concentrated under vacuum. The crude product was purified by silica gel chromatography (eluting with 3:2 ethyl acetate/petroleum ether) to give (2S) -5- [2- (5-fluoro-2-methoxyphenyl) -2-hydroxyacetamido as a yellow solid]-6- [ [ (1R,3R) -3- (methoxycarbonyl) cyclohexyl]Amino group]-methyl 2-methyl-1, 2,3, 4-tetrahydroquinoline-1-carboxylate (180mg, 81%). LCMS (ES, M/z):558[ M + H]⁺。

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.

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

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')

Reacting (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]A solution of quinoline-6-carboxylic acid methyl ester (120mg,0.22mmol) and LiOH (16mg,0.67mmol) in tetrahydrofuran (2.0mL), methanol (2.0mL) and water (2.0mL) was stirred at 25 ℃ overnight. The resulting mixture was concentrated under vacuum. The crude product was passed through preparative HPLC (column, Xbridge Prep C18OBD column, 19X150 mm,5 um; mobile phase, A: water (containing 10mmol/L NH)₄HCO₃) And B: ACN (15.0% to 29.0% in 14 min); detector, UV 220/254 nm). The product was separated by chiral preparative HPLC (column, CHIRALPAK IE, 2X25cm,5 um; mobile phase, A: Hex (containing 0.1% FA) and B: ethanol (50.0% ethanol over 12 min); detector, UV 220/254 nm). The product fractions were concentrated to give (1R,3R) -3- [ (7S) -2- [ (R) - (5-fluoro-2-methoxyphenyl) (hydroxy) methyl as a white solid]-6- (methoxycarbonyl) -7-methyl-3H, 6H,7H,8H, 9H-imidazo [4,5-f]Quinolin-3-yl]Cyclohexane-1-carboxylic acid (23.6mg, 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 (23.8mg, 20%). Stereoisomeric purity by HPLC: column: CHIRALPAK IE-3, column size: 0.46x5 cm; 3 μm; mobile phase: hex (0.1% FA): EtOH 50:50, flow: 1.0 ml/min. Conversion as determined by HPLC analysisThe purity of the compound was estimated to be greater than 99%.

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]⁺。

second eluting isomer (1'):¹H-NMR(CD₃OD,400MHz)δ(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＝6.4Hz,3H)。LCMS(ES,m/z):526[M+H]⁺。

example 1 preparation of hydrochloric acid addition salts of form b-A

900mg 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 16.5mL 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 at the third addition, the solid was still present. The slurry was stirred in a 50 ℃ 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 908mg (94% yield). LCMS (ES, M/z) 526[ M + H ]]⁺The samples were further characterized by XRPD, DSC, and TGA. The results show that the sample is crystalline according to XRPD, corresponding to the HCL salt form a.

Example 1 preparation of c-hydrochloric acid addition salt: type B

20mg of the hydrochloric acid addition salt form A 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 are dissolved in 0.4mL of a 1:1 mixture of dichloromethane and isopropanol. The solution was heated to 50 ℃ and then cooled to 5 ℃ at a rate of 0.1 ℃ per minute, then stirred at 5 ℃ overnight. The solution was then warmed to room temperature and precipitation was observed after a portion of the solvent was slowly evaporated without assistance. The precipitate was collected by centrifugation, dried at 40 ℃ under vacuum and subsequently characterized by XRPD, DSC and TGA. The results show that by XRPD the sample is crystalline, conforming to the hydrochloric acid addition salt form B.

Example 1 preparation of d-hydrochloric acid addition salt: c type

20mg of the hydrochloric acid addition salt form A 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 0.4mL of a 9:1 mixture of 1, 4-dioxane and water. The solution was heated to 50 ℃ and then cooled to 5 ℃ at a rate of 0.1 ℃ per minute, then stirred at 5 ℃ overnight. The solution was then warmed to room temperature and precipitation was observed after a portion of the solvent was slowly evaporated without assistance. The precipitate was collected by centrifugation, dried at 40 ℃ under vacuum and subsequently characterized by XRPD, DSC and TGA. The results show that by XRPD the sample is crystalline, conforming to the hydrochloric acid addition salt form C.

Example 1. preparation of the e-p-toluenesulfonic acid addition salt: type A

20mg 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 0.4mL 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 at 40 ℃ under vacuum, followed by XRPD, DSC, TGA and¹h NMR was carried out for characterization. The results show that by XRPD the sample is crystalline, conforming to the p-toluenesulfonic acid addition salt form a.

Example 1 preparation of p-toluenesulfonic acid addition salts of form f-B

300.6mg 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 4.0mL acetonitrile in a 20mL vialIn (b), a suspension is formed. 107.34mg of p-toluenesulfonic acid were dissolved in 1.5mL of acetonitrile and slowly added to the suspension. The suspension was then heated to 50 ℃ and subsequently cooled to 5 ℃ at a rate of 0.1 ℃ per minute. After stirring overnight at 5 ℃, the resulting precipitate was collected by centrifugation and dried under vacuum at 40 ℃ to give 231.8mg of a white powder. By XRPD, DSC, TGA and¹h NMR characterised the solid and indicated that the sample was crystalline by XRPD and conformed to the p-toluene sulfonic acid addition salt form B.

Example 1 preparation of g-benzenesulfonic acid addition salt: type A

20mg 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 0.4mL ethyl acetate and mixed with 1 equivalent of benzenesulfonic acid. The resulting solution was stirred at room temperature for 3 days. After no precipitate was observed at the three day mark, the solution was heated to 50 ℃ and slowly cooled to 5 ℃ at a rate of 0.1 ℃ per minute. After stirring the solution at 5 ℃ overnight, a solid precipitate was observed. The resulting solid was collected by centrifugation, dried at 40 ℃ under vacuum, followed by XRPD, DSC, TGA and¹h NMR was carried out for characterization. The results show that by XRPD the sample is crystalline, conforming to the benzenesulfonic acid addition salt form a.

Example 1 preparation of h-sulfuric acid addition salt: type A

20mg 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 are dissolved in 0.4mL of ethyl acetate and mixed with 1 equivalent of sulfuric acid. The resulting solution was stirred at room temperature for 3 days. After no precipitate was observed at the three day mark, the solution was heated to 50 ℃ and slowly cooled to 5 ℃ at a rate of 0.1 ℃ per minute. After stirring the solution at 5 ℃ overnight, a solid precipitate was observed. The resulting solid was collected by centrifugation, dried at 40 ℃ under vacuum and subsequently characterized by XRPD, DSC and TGA. The results show that by XRPD the sample is crystalline, conforming to the form a sulfuric acid addition salt.

Example 1 preparation of crystalline free form of i-A

Mixing (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 (5.00g,9.51mmol) was completely dissolved in hot ethyl acetate (67mL) at reflux. Turbidity appeared after a short cooling time of a few minutes. The hot slurry was slowly cooled to ambient temperature and then stirred for 1 hour. The slurry was then cooled to 0 ℃ and stirred in an ice bath for 2 hours, then filtered and the solid washed with cold ethyl acetate. 3.70g (74% recovery) of a white powder with increased purity after drying was isolated. LCMS (ES, M/z) 526[ M + H ]]⁺. The samples were further characterized by XRPD, DSC, and TGA. These results indicate that by XRPD, the sample is crystalline, consistent with form a free form.

Example 2 kinetic solubility assessment

The solubility of the HCl salt type A, the tosylate salt type B and the free form type A in water, SGF, FaSSIF and FeSSIF was measured at 37 ℃ with a solids loading of about 10mg/mL (calculated as free base). All solubility samples were kept spinning at 25rpm at 37 ℃ and sampled at 1h, 4h and 24h, respectively. 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 fig. 7.

EXAMPLE 3 HTRF Biochemical assay of CBP and BRD4 Activity

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

the assay was performed in a final volume of 6. mu.L in assay buffer containing 50mM Hepes (pH 7.5, (0.5M Hepes, pH 7.5 solution; Teknova H1575)), 0.5mM GSH, 0.01% BGG (0.22. mu.M filtered, Sigma, G7516-25G), 0.005% BSA (0.22. mu.M filtered, EMD Mil)lipore Corporation,126575) and 0.01% Triton X-100(Sigma, T9284-10L). Nanoliters of 10-point, 3-fold serial dilutions in DMSO were pre-dispensed into 1536 assay plates (Corning, #3724BC) at final assay concentrations ranging from 33 μ M to 1.7nM, highest to lowest dose respectively. mu.L of 2X protein and 3. mu.L of 2X peptide ligand were added to the assay plate (pre-imprinted with compound). The plates were incubated at room temperature for various times before measuring the signal. In the PHERAStar plate reader (BMG, equipped with HTRF optical module [337/520/490 ]]) Or TR-FRET (time resolved fluorescence resonance energy transfer) on an Envision plate reader (Perkinelmer, equipped with a TRF laser unit, a TRF double mirror D400/D505 and emission filters 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 [ (] 520nm fluorescence/490 nm fluorescence) [ ] 10000 ], AveLow [ (] 32) ] the average TR-FRET ratio for the no enzyme control, and AveHigh [ (] 32) ] the average TR-FRET ratio for the DMSO control. IC50 values were determined by curve fitting using a standard 4-parameter logistic fitting algorithm, which is contained 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 [% FLU-AveLow)/(Avehigh-AveLow) ], where FLU [% measured fluorescence, AveLow [% average fluorescence of no enzyme control (n [ ] 32), Avehigh [ ] average fluorescence of DMSO control (n [ ] 32). IC50 values were determined by curve fitting using a standard 4-parameter logistic fitting algorithm, which is contained in the Activity Base software package: IDBS XE Designer Model 205. The data were fitted using the Levenburg Marquardt algorithm. IC observed for CBP inhibition by Compound 1₅₀Measured between 0.001 μ M and 0.01 μ M, IC for BRD4 inhibition₅₀The measurement values are between 1.0. mu.M and 1000. mu.M. IC observed for CBP inhibition by Compound 2₅₀Measured between 0.001 μ M and 0.01 μ M, IC for BRD4 inhibition₅₀The measurement values are between 1.0. mu.M and 1000. mu.M.

Example 4-Compound 1 and Compound 1' demonstrateIn vitro Activity against CBP

Comprising compound 1 and compound 1':

the potency and selectivity of the included CBP/P300 inhibitor compounds was determined in a biochemical time-resolved fluorometric assay using GST fusions of the bromodomains of CBP and BRD 4. Briefly, CBP inhibitors were pre-dispensed into 1536 assay plates at final assay concentrations ranging from 33 μ M to 1.7 nM. Proteins and ligands were added to the compounds to a final concentration of 2.5nM of CBP or BRD4 (N-terminal GST-CREBP (1081-CREBBP 1197), BRD4 tandem domain) and 25nM of tetraacetylated H3 peptide plates and incubated for 4 hours. Data are reported as percent inhibition compared to control wells. IC (integrated circuit)₅₀Values were determined by curve fitting of a standard 4-parameter logistic fitting algorithm. Under these conditions, compound 1 was identified as a potent inhibitor of CBP, where IC₅₀<2nM (N ═ 16). In a similar assay, the potency of BRD4 was determined and Compound 1 showed<IC of 500nM₅₀(N ═ 15) indicates that>200 times selectivity.

The selectivity of compound 1 was evaluated in a screening assay for kinase inhibition and BRD binding. Compound 1 in KINOMEscan^TMThe human kinase and disease-associated mutant variants evaluated in the screen showed no to low binding affinity. A small set of 10 BRDs representing different branched or bromodomain trees were tested using AlphaScreen. Compound 1 was not active on 8 of the 10 bromine domains investigated. IC of Compound 1 against bromodomains of CREBP and BRD4 (concatemeric BD1/BD2)₅₀Values of 0.1. mu.M and>10 μ M, confirming the high selectivity of compound 1 for CBP.

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:

[0001]compound (I)	CBP(IC50)	BRD4(IC50)/CBP(IC50) Selectivity ratio of
			1	<10nM	>266
1′	<20nM	>76

In the HTRF biochemical assay of example 3, both compound 1 and compound 1' inhibited efficiently (e.g., IC)₅₀<100nM) CBP, and using this assay, compound 1 was approximately 3.5 times more selective for CBP inhibition than for BRD4 inhibition.

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

1.a compound of formula (I):

and non-amorphous solid forms of salts thereof.

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

and salts thereof.

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

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

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 (° 2 θ ± 0.2) selected from the group consisting of: 7.27, 8.98, 10.60, 15.60 and 23.93.

6. The solid form of clause 4, wherein the hydrochloric acid addition salt form is form B, characterized by an X-ray powder diffraction pattern comprising at least three peak positions (° 2 θ ± 0.2) selected from the group consisting of: 10.23, 18.72, 23.03, 24.77 and 28.03.

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 (° 2 θ ± 0.2) selected from the group consisting of: 7.05, 19.84, 21.09, 24.98 and 31.44.

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

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 (° 2 θ ± 0.2) selected from the group consisting of: 6.59, 13.20, 14.46, 18.00 and 21.74.

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 (° 2 θ ± 0.2 degrees) selected from the group consisting of: 7.10, 9.15, 15.08, 16.19, 17.25, 18.31 and 21.13.

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

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 (° 2 θ ± 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.

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

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 (° 2 θ ± 0.2) selected from the group consisting of: 8.24, 9.98, 13.58, 16.87, 18.78, 20.00 and 25.52.

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

16. A method of preparing a solid form of the compound of clause 1, comprising:

suspending the free base, amorphous compound of formula (I) in an acidic organic solvent or mixture of organic solvents to form a solution; heating the solution; and cooling the solution.

17. A method of preparing a solid form of the compound of clause 15, comprising:

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

heating the solution; and

the solution was cooled, and then the precipitated solid matter was filtered and washed.

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

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.

20. A method of treating, preventing, inhibiting, or eliminating a disease or disorder 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.

21. The solid form of any one of clauses 1-15 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.

22. Use of the solid form of any one of clauses 1-15 or the composition of clause 18 for 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:

1.a solid form of a compound of formula (II):

wherein said solid form is a hydrochloric acid addition salt, and wherein said hydrochloric acid addition salt is the hydrochloric acid addition salt form A.

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 (° 2 θ ± 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.

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

4. The solid form of any of clauses 1-3, wherein the solid form is characterized by a weight loss of about 1.1% at a temperature of up to 170 ℃ as measured by thermogravimetric analysis.

5. The solid form of any one of clauses 1-4, wherein the solid form is an anhydrate.

6. The solid form of any one of clauses 1-5, wherein the solid form is hygroscopic.

7. The solid form of any one of clauses 1-6, wherein the solid form is stable at a temperature of up to 40 ℃ and a relative humidity of up to 75% for at least two weeks.

8. A process for preparing a solid form of compound (II):

the method comprises the following steps:

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

adding hydrochloric acid to the solution;

wherein said solid form is a hydrochloric acid addition salt, and wherein said hydrochloric acid addition salt is the hydrochloric acid addition salt form A.

9. The method of clause 8, wherein the organic solvent is ethyl acetate.

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

11. The method of any of clauses 8-10, wherein the method further comprises heating the solution.

12. The method of any of clauses 8-11, wherein the method further comprises cooling the solution.

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

14. 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 solid form of any one of clauses 1-7 or the pharmaceutical composition of clause 13.

15. A method of treating, preventing, inhibiting, or eliminating a disease or disorder 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 1-7 or the pharmaceutical composition of clause 13.