Solid forms of rock inhibitors

Abstract

The present invention relates to single enantiomers and crystalline forms of Rho-associated protein kinase (ROCK) inhibitors useful for the treatment of fibrotic diseases, such as fibrostenosis Crohn's disease.

Description

Technical Field

[0001] The present invention relates to single enantiomers and crystalline forms of 4-(aminomethyl)-N-(3-fluoropyridin-4-yl)-3-{3-[(4-{[(2-oxoxolan-3-yl)methyl]mercapto}phenyl)carbamoyl]phenyl}benzamide (Compound A). Compound A is a Rho-associated protein kinase (ROCK) inhibitor and can be used to treat fibrotic diseases, such as fibrostenotic Crohn's disease. Background Art

[0002] 4-(Aminomethyl)-N-(3-fluoropyridin-4-yl)-3-{3-[(4-{[(2-oxoxolan-3-yl)methyl]thio}phenyl)carbamoyl]phenyl}benzamide (Compound A) is a potent Rho-associated protein kinase (ROCK) inhibitor useful for treating fibrotic diseases (see WO2014 / 118133). In particular, Compound A has been designed to act specifically at fibrotic sites in the gastrointestinal (GI) tract and, if absorbed into the bloodstream, to be rapidly degraded by enzyme-mediated metabolism.

[0003]

[0004] Crohn's disease is a chronic inflammatory bowel disease that affects 1.5 million people worldwide, with more than 70,000 new cases diagnosed each year. Up to 50% of Crohn's disease patients may develop significant fibrosis and stricture formation within ten years of diagnosis; this type of Crohn's disease-related fibrosis is called fibrostenotic Crohn's disease.

[0005] Current treatment for fibrotic strictures of the GI tract is primarily surgical, as there are no drugs specifically approved for fibrosis and fibrosis can progress despite intervention with anti-inflammatory therapies.

[0006] The recurrence rate after surgical intervention is high, with more than 50% of patients requiring further surgery within 10 years, and many within 12 months. As a result, patients experience a progressive loss of GI function, and repeated resections can lead to serious health complications, such as short bowel syndrome.

[0007] Preclinical data showed that compound A exhibited strong anti-fibrotic therapeutic effects in multiple animal models of inflammatory bowel disease.

[0008] The free base of Compound A has low stability.

[0009] It is an object of certain embodiments of the present invention to provide a stable crystalline form of Compound A. It is an object of certain embodiments of the present invention to provide a crystalline form of Compound A that is more stable than other crystalline forms.

[0010] Certain embodiments of the present invention meet some or all of the above objectives. Summary of the invention

[0011] In a first aspect of the present invention, there is provided the S-enantiomer (SA) of compound A: It is substantially in enantiomerically pure form, or a pharmaceutically acceptable salt thereof. The S-enantiomer of compound A may be in the form of a free base. The S-enantiomer of compound A may be in the form of a pharmaceutically acceptable salt. The salt may be in the form of a crystalline salt.

[0012] The inventors have found that the S-enantiomer of compound A exhibits lower inhibition than the R-enantiomer against a variety of alternative targets. In particular, the S-enantiomer of compound A exhibits lower inhibition than the R-enantiomer against a variety of kinases other than ROCK. This is particularly surprising because, to date, the inventors have not detected a statistically significant difference between the activity of the S-enantiomer and the R-enantiomer of compound A against the kinases ROCK1 or ROCK2.

[0013] Furthermore, the S-enantiomer is less active than the R-enantiomer in a number of assays used to identify undesirable off-target activities that could have adverse effects in humans.

[0014] When the present invention relates to a pharmaceutically acceptable salt of Compound A, the salt is generally an acid addition salt. Suitable acid addition salts are formed from acids which form non-toxic salts. Examples include acetate, aspartate, benzoate, benzenesulfonate, bicarbonate / carbonate, bisulfate / sulfate, borate, camphorsulfonate, citrate, edisylate, ethanesulfonate, formate, fumarate, glucoheptonate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, maleate, malonate, methanesulfonate, methylsulfate, naphthylate, 1,5-naphthalenedisulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / hydrogenphosphate / dihydrogenphosphate, sucrose, stearate, succinate, tartrate, toluenesulfonate, and trifluoroacetate.

[0015] The salt may be selected from hydrochloride and succinate. The inventors have found that succinate and hydrochloride salts of compound A form crystals that are more stable than the free base. The salt may be succinate. The salt may be hydrochloride. The inventors have found that the crystalline form of the hydrochloride salt of compound A is particularly stable.

[0016] In a second aspect of the present invention, there is provided a crystalline salt form of Compound A:

[0017]

[0018] The free base of Compound A has low chemical stability, especially towards ester hydrolysis. The present inventors have found that the salt form of Compound A exhibits improved stability relative to the free base.

[0019] The present invention relates to a pharmaceutically acceptable salt of compound A. The salt is usually an acid addition salt.

[0020] Suitable acid addition salts are formed from acids that form non-toxic salts. Examples include acetate, aspartate, benzoate, benzenesulfonate, bicarbonate / carbonate, bisulfate / sulfate, borate, camphorsulfonate, citrate, edisylate, ethanesulfonate, formate, fumarate, glucoheptonate, gluconate, glucuronate, hexafluorophosphate, hyaluronate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, maleate, malonate, methanesulfonate, methylsulfate, naphthoate, 1,5-naphthalenedisulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / hydrogenphosphate / dihydrogenphosphate, sucrose, stearate, succinate, tartrate, toluenesulfonate and trifluoroacetate.

[0021] The salt may be selected from hydrochloride and succinate. The inventors have found that succinate and hydrochloride salts of compound A form crystals that are more stable than the free base. The salt may be succinate. The salt may be hydrochloride. The inventors have found that the crystalline form of the hydrochloride salt of compound A is particularly stable.

[0022] The crystalline salt form may be a solvate. The crystalline salt form may be a hydrate. The crystalline salt form may not be a solvate. The crystalline salt form may not be a hydrate.

[0023] Compound A contains a single chiral center located at the point of attachment of the furanone ring to the rest of the molecule. In the crystalline form of the first aspect of the invention, Compound A can be a mixture of two enantiomers. Compound A can be a racemic mixture of two enantiomers. Compound A can be substantially in the form of a single enantiomer.

[0024] A single enantiomer may be the S-enantiomer (SA):

[0025]

[0026] A single enantiomer may be the R-enantiomer (RA):

[0027]

[0028] The word "substantially" may mean that compound A has an enantiomeric excess of 90% or more. The word "substantially" generally means that compound A has an enantiomeric excess of 95% or more. This may mean that compound A has an enantiomeric excess of 98% or more, 99% or more, or 99.5% or more. Throughout the specification, when compound A is described as a single enantiomer (R or S), unless the enantiomeric excess is otherwise indicated, it is intended to mean that it is in substantially enantiomerically pure form within the meaning of this paragraph.

[0029] The enantiomer may be the faster eluting enantiomer in chiral HPLC performed using a column comprising silica coated with cellulose-tris(3,5-dimethylphenylcarbamate), such as a Chiralcel OD-3 column. The enantiomer may be the slower eluting enantiomer in chiral HPLC performed using a column comprising silica coated with cellulose-tris(3,5-dimethylphenylcarbamate), such as a Chiralcel OD-3 column. The solvent system used as the mobile phase in HPLC may be 28% solvent mixture B in solvent mixture A, wherein solvent mixture A is heptane containing 0.05% diethylamine; and solvent mixture B is a 4:1 mixture of isopropanol and acetonitrile, also containing 0.05% diethylamine.

[0030] The salt may be a hydrochloride salt of a single enantiomer of compound A, characterized in that when K α2 / K α1 The crystalline salt form has an XRPD pattern having at least two peaks at 2θ selected from 16.2±0.2, 22.7±0.2, 23.3±0.2, 24.0±0.2, 24.9±0.2 and 25.4±0.2 when measured with Cu radiation at a ratio of 0.5. The salt may be a hydrochloride salt of a single enantiomer of compound A, characterized in that when using K α2 / K α1 The crystalline salt form has an XRPD pattern having at least four peaks at 2θ selected from 16.2±0.2, 22.7±0.2, 23.3±0.2, 24.0±0.2, 24.9±0.2 and 25.4±0.2 when measured with Cu radiation at a ratio of 0.5. The salt may be a hydrochloride salt of a single enantiomer of compound A, characterized in that when K α2 / K α1The crystalline salt form has an XRPD pattern having peaks at 2θ selected from 16.2±0.2, 22.7±0.2, 23.3±0.2, 24.0±0.2, 24.9±0.2 and 25.4±0.2 when measured using Cu radiation with a K ratio of 0.5. α2 / K α1 The crystalline form may have substantially the same Figure 1 The XRPD pattern shown. This crystalline form is referred to as HCl salt form I in this specification. This crystalline form is a trihydrate. The single enantiomer can be the S enantiomer. The single enantiomer can be the R enantiomer.

[0031] The salt may be a hydrochloride salt of a single enantiomer of compound A, characterized in that when K α2 / K α1 The crystalline salt form has an XRPD pattern having at least two peaks at 2θ selected from 13.6±0.2, 14.4±0.2, 14.5±0.2, 16.2±0.2 and 16.5±0.2 when measured with Cu radiation at a ratio of 0.5. The salt may be a hydrochloride salt of a single enantiomer of compound A, characterized in that when using K α2 / K α1 The crystalline salt form has an XRPD pattern having at least four peaks at 2θ selected from 13.6±0.2, 14.4±0.2, 14.5±0.2, 16.2±0.2 and 16.5±0.2 when measured with Cu radiation at a ratio of 0.5. The salt may be a hydrochloride salt of a single enantiomer of compound A, characterized in that when using K α2 / K α1 The crystalline salt form has an XRPD pattern having peaks at 2θ selected from 13.6±0.2, 14.4±0.2, 14.5±0.2, 16.2±0.2 and 16.5±0.2 when measured with Cu radiation at a K ratio of 0.5. α2 / K α1 The crystalline form may have substantially the same Figure 3 The XRPD pattern shown. This crystalline form is referred to as HCl salt form II in this specification. This crystalline form is an anhydrate. The single enantiomer can be the S enantiomer. The single enantiomer can be the R enantiomer.

[0032] The salt may be a succinate salt of a single enantiomer of compound A, characterized in that when K α2 / K α1The crystalline salt form has an XRPD pattern having at least two peaks at 2θ selected from 18.2±0.2, 18.6±0.2, 19.1±0.2, 21.4±0.2, 23.0±0.2, 24.1±0.2 and 25.8±0.2 when measured with Cu radiation at a ratio of 0.5. The salt may be a succinate salt of a single enantiomer of Compound A, characterized in that when using K α2 / K α1 The crystalline salt form has an XRPD pattern having at least four peaks at 2θ selected from 18.2±0.2, 18.6±0.2, 19.1±0.2, 21.4±0.2, 23.0±0.2, 24.1±0.2 and 25.8±0.2 when measured with Cu radiation at a ratio of 0.5. The salt may be a succinate salt of a single enantiomer of Compound A, characterized in that when using K α2 / K α1 The crystalline salt form has an XRPD pattern having peaks at 2θ selected from 18.2±0.2, 18.6±0.2, 19.1±0.2, 21.4±0.2, 23.0±0.2, 24.1±0.2 and 25.8±0.2 when measured using Cu radiation with a K ratio of 0.5. α2 / K α1 The crystalline form may have substantially the same Figure 2 The XRPD pattern shown. This crystalline form is referred to as succinate form I in this specification. The single enantiomer may be the S enantiomer. The single enantiomer may be the R enantiomer.

[0033] HCl Salt Form I, HCl Salt Form II, and Succinate Form I are all chemically stable crystalline materials and are more stable than the free base of Compound A. HCl Salt Form I and HCl Salt Form II are more stable than Succinate Form I. HCl Salt Form II is particularly beneficial because it is more crystalline than HCl Salt Form I and is also more soluble at low pH.

[0034] In the third aspect of the present invention, a pharmaceutical preparation is provided, which comprises the crystalline salt form of the first aspect or the compound A of the second aspect or a pharmaceutically acceptable salt thereof.

[0035] In a fourth aspect of the present invention, there is provided a crystalline salt form of the first aspect or Compound A of the second aspect or a pharmaceutically acceptable salt thereof for use in medical treatment.

[0036] In the fifth aspect of the present invention, there is provided a crystalline salt form of the first aspect or compound A or a pharmaceutically acceptable salt thereof of the second aspect for use in treating a disease.

[0037] In the sixth aspect of the present invention, a method for treating a disease is provided, which comprises administering a therapeutically effective amount of the crystalline salt form of the first aspect or the compound A or a pharmaceutically acceptable salt thereof of the second aspect to a subject in need thereof.

[0038] In the seventh aspect of the present invention, there is provided use of the crystalline salt form of the first aspect or the compound A or a pharmaceutically acceptable salt thereof of the second aspect in the preparation of a medicament for treating a disease.

[0039] The disease may be a fibrotic disease.

[0040] The disease may be a disease of the gastrointestinal system.

[0041] The disease may be inflammatory bowel disease.

[0042] The disease may be Crohn's disease, such as fibrostenotic Crohn's disease

[0043] The disease may be cancer, such as colorectal cancer.

[0044] The disease may be a skin disease such as keloid or scleroderma.

[0045] The disease may be an eye disease.

[0046] In an eighth aspect of the present invention, there is provided a method for preparing a single enantiomer of Compound A, HCl crystalline salt form II, the method comprising:

[0047] a) suspending a sample of the HCl crystalline salt Form I of a single enantiomer of Compound A in a solvent to form a suspension;

[0048] b) Filtering the suspension to obtain HCl crystalline salt Form II.

[0049] The solvent may be a mixture of solvents.

[0050] The solvent may be a polar aprotic organic solvent. The solvent may include a polar aprotic organic solvent. The solvent may be water. The solvent may include water. The solvent may be a polar protic organic solvent. The solvent may include a polar protic organic solvent.

[0051] The solvent may be a mixture of a polar aprotic organic solvent and a solvent selected from a polar protic organic solvent and water. The solvent may be a mixture of a polar aprotic organic solvent and water.

[0052] Suitable polar aprotic organic solvents include dimethyl sulfoxide (DMSO), acetone, acetonitrile, ethers (eg THF, diglyme, ethylene glycol dimethyl ether, tert-butyl methyl ether), esters (eg ethyl acetate).

[0053] Suitable polar protic organic solvents include ethanol, methanol, isopropanol, ethylene glycol.

[0054] The solvent may comprise acetone. The solvent is preferably a mixture of acetone and water. The inventors have found that using a mixture of acetone and water provides a cleaner product than other possible solvent systems.

[0055] The ratio of polar aprotic organic solvent (e.g. acetone) to water may be in the range of 1:1 to 50:1 acetone:water by volume. The ratio of polar aprotic organic solvent (e.g. acetone) to water may be in the range of 10:1 to 30:1 acetone:water by volume. The ratio of polar aprotic organic solvent (e.g. acetone) to water may be in the range of 15:1 to 25:1 acetone:water by volume.

[0056] The single enantiomer may be the S enantiomer. The single enantiomer may be the R enantiomer.

[0057] Step a) generally comprises stirring the mixture. The mixture may be stirred for more than 1 hour. The mixture may be stirred for more than 5 hours. The mixture may be stirred for more than 12 hours. The mixture may be stirred for less than 48 hours. The mixture may be stirred for less than 24 hours.

[0058] Step a) is generally carried out at a temperature ranging from 5°C to 40°C. Step a) is generally carried out at a temperature ranging from 15°C to 30°C.

[0059] The method may further comprise: step c) drying the HCl salt form II, for example under vacuum. Step c) may be performed at a temperature in the range of 30°C to 60°C. Step c) may be performed for more than 30 minutes. Step c) may be performed for less than 6 hours, for example less than 3 hours. BRIEF DESCRIPTION OF THE DRAWINGS

[0060] Embodiments of the present invention are further described below with reference to the accompanying drawings, in which:

[0061] Figure 1 is the XRPD spectrum of HCl salt form I

[0062] Figure 2 is the XRPD spectrum of succinate form I

[0063] Figure 3 is the XRPD spectrum of HCl salt form II DETAILED DESCRIPTION

[0064] The present invention relates to pharmaceutically acceptable salts of Compound A. For a review of suitable salts, see "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" by Stahl and Wermuth (Wiley-VCH, Weinheim, Germany, 2002).

[0065] The pharmaceutically acceptable acid addition salts of Compound A can be prepared by one or more of the following two methods:

[0066] By reacting compound A with a desired acid;

[0067] One salt of Compound A is converted to another salt by reaction with a suitable acid or base or by passing through a suitable ion exchange column.

[0068] These reactions are usually carried out in solution. The resulting salt may precipitate and be collected by filtration, or it may be recovered by evaporation of the solvent. The degree of ionization of the resulting salt may vary from fully ionized to almost non-ionized.

[0069] The term "solvate" is used herein to describe a molecular complex comprising a compound of the invention and one or more pharmaceutically acceptable solvent molecules (eg, ethanol). When the solvent is water, the term "hydrate" is used.

[0070] It is known in the art that an X-ray powder diffraction pattern with one or more measurement errors can be obtained, depending on the measurement conditions (such as equipment, sample preparation or the machine used). In particular, it is well known that the intensity in the X-ray powder diffraction pattern can fluctuate depending on the measurement conditions and sample preparation. For example, a technician in the field of X-ray powder diffraction will recognize that the relative intensity of the peak can vary depending on the orientation of the test sample and the type and setting of the instrument used. The technician will also recognize that the position of the reflection will be affected by the precise height of the sample in the diffractometer and the zero calibration of the diffractometer. The surface flatness of the sample may also have a small effect. Therefore, it will be understood by those skilled in the art that the diffraction pattern data given herein should not be interpreted as absolute, and any crystalline form that provides a powder diffraction pattern substantially the same as those disclosed herein falls within the scope of the present disclosure (for more information, see "Jenkins, R & Snyder, RL 'Introduction to X-Ray Powder Diffractometry' John Wiley & Sons, 1996").

[0071] The term "stable" may refer to chemical stability or physical stability. In particular, the free base compound A is chemically unstable, while the crystalline form of the present invention is chemically stable for 7 days at 40°C and 75% relative humidity, and / or chemically stable for 7 days at 60°C and ambient humidity. The crystalline form of the present invention is chemically stable for 3 months at 40°C and 75% relative humidity, and / or chemically stable for 3 months at 25°C and 60% relative humidity.

[0072] For the above-mentioned compounds of the present invention, the dosage will certainly vary with the compound used, mode of administration, desired treatment and indicated disease. For example, if the compound of the present invention is administered orally, the daily dosage of the compound of the present invention can be in the range of 0.01 microgram / kilogram body weight (μg / kg) to 100 milligrams / kilogram body weight (mg / kg).

[0073] The crystalline salt form or compound of the present invention can be used alone, but will usually be administered in the form of a pharmaceutical composition, wherein the crystalline salt form or compound of the present invention or a pharmaceutically acceptable salt thereof is combined with a pharmaceutically acceptable adjuvant, diluent or carrier. Conventional procedures for selecting and preparing suitable pharmaceutical formulations are described in, for example, "Pharmaceuticals-The Science of Dosage Form Designs", ME Aulton, Churchill Livingstone, 1988.

[0074] Depending on the mode of administration of the crystalline salt form or compound of the present invention, the pharmaceutical composition for administering the crystalline salt form or compound of the present invention will preferably contain 0.05 to 99% w (weight percentage) of the crystalline salt form or compound of the present invention, more preferably 0.05 to 80% w of the crystalline salt form or compound of the present invention, still more preferably 0.10 to 70% w of the crystalline salt form or compound of the present invention, and even more preferably 0.10 to 50% of the crystalline salt form or compound of the present invention, all weight percentages being based on the total composition.

[0075] The pharmaceutical composition can be applied topically (e.g., to the skin or eyes) in the form of, for example, a cream, gel, lotion, solution, suspension, or systemically, for example, orally in the form of a tablet, capsule, syrup, powder or granules; rectally in the form of a suppository; or by inhalation in the form of an aerosol.

[0076] For oral administration, the crystalline salt forms or compounds of the present invention can be mixed with adjuvants or carriers, such as lactose, sucrose, sorbitol, mannitol; starches, such as potato starch, corn starch or pullulan; cellulose derivatives; binders, such as gelatin or polyvinyl pyrrolidone; and / or lubricants, such as magnesium stearate, calcium stearate, polyethylene glycol, wax, paraffin, etc., and then compressed into tablets. If coated tablets are required, the core prepared as described above can be coated with a concentrated sugar solution, which can contain, for example, gum arabic, gelatin, talc and titanium dioxide. Alternatively, the tablets can be coated with a suitable polymer dissolved in a volatile organic solvent.

[0077] In order to prepare soft gelatin capsules, the crystalline salt form of the present invention or compound can be mixed with, for example, vegetable oil or polyethylene glycol. Hard gelatin capsules can contain salt form particles prepared using the above-mentioned excipients for tablets. In addition, the crystalline salt form of the present invention or the liquid or semisolid preparation of the compound can be filled into hard gelatin capsules. Liquid preparations for oral use can be in the form of syrup or suspension, and the surplus is a mixture of sugar and ethanol, water, glycerol and propylene glycol. Optionally, this liquid preparation can contain coloring agent, flavoring agent, sweetener (such as saccharin), preservative and / or carboxymethyl cellulose as thickener or other excipients known to those skilled in the art.

[0078] According to well-known medical principles, the dosage size of the crystalline salt form or compound of the present invention for therapeutic purposes will naturally vary depending on the nature and severity of the condition, the age and sex of the animal or patient and the route of administration.

[0079] The dosage level, dosing frequency and duration of treatment of the crystalline salt forms or compounds of the present invention are expected to vary depending on the formulation and the clinical indication, age and comorbid medical conditions of the patient.

[0080] Throughout the description and claims of this specification, the words "comprise" and "comprising" and their variations mean "including but not limited to", and they are not intended to (and do not) exclude other parts, additives, ingredients, integers or steps. Throughout the description and claims of this specification, the singular includes the plural unless the context requires otherwise. In particular, where the indefinite article is used, the specification should be understood as contemplating plurality as well as singularity, unless the context requires otherwise.

[0081] Features, integers, characteristics, compounds, chemical moieties or groups described in conjunction with a particular aspect, embodiment or example of the invention are to be understood to be applicable to any other aspect, embodiment or example described herein unless incompatible therewith. All features disclosed in this specification (including any accompanying claims, abstract and drawings), and / or all steps of any method or process so disclosed, may be combined in any combination, except combinations in which at least some of such features and / or steps are mutually exclusive. The invention is not limited to the details of any foregoing embodiments. The invention extends to any novel one or any novel combination of the features disclosed in this specification (including any accompanying claims, abstract and drawings), or to any novel one or any novel combination of the steps of any method or process so disclosed.

[0082] The reader's attention is directed to all papers and documents filed concurrently with or prior to this specification in connection with this application, which are disclosed to the public with this specification, and the contents of all these papers and documents are incorporated herein by reference.

[0083] Example

[0084] Example 1 - Formation of racemic compound 4

[0085] Preparation of compound 3-

[0086]

[0087] To a suspension of compound 2 (98.0 g, 999 mmol, 1.00 equiv) in THF (tetrahydrofuran; 980 mL) at 25°C under N2 was added TEA (triethylamine; 101 g, 999 mmol, 139 mL, 1.00 equiv) and compound 1 (233 g, 1.50 mol, 1.50 equiv). The mixture was stirred at 25°C under N2 for 6 h. LCMS showed that compound 2 was consumed and the desired MS (RT = 0.592 min) was detected. The reaction was filtered and the solid was washed with petroleum ether (1.00 L) to wash off the dark color. The filter cake was concentrated to give racemic compound 3 (389 g, 1.53 mol, 76.3% yield, 99.3% purity) as a yellow solid. LCMS: Product: RT = 0.592 min, m / z = 254.0 (M+H) +

[0088] 1H NMR: 400MHz, DMSOδ / ppm: 8.15(d,J=8.8Hz,2H),7.56(d,J=8.8Hz,2H),4.35-4.30(m,1H),4.20-4.15(m,1H ),3.54(dd,J=4.8,4.4Hz,1H),3.31-3.26(m,1H),3.12-3.05(m,1H),2.39-2.34(m,1H),2.11-2.00(m,1H).

[0089] Preparation of compound 4-

[0090]

[0091] To a solution of racemic compound 3 (131 g, 512 mmol, 99.3% purity, 1.00 equiv) in MeOH (366 mL), THF (366 mL) and DMA (dimethylacetamide; 52.0 mL) was added Pd / C (41.8 g, 512 mmol, 10.0% purity, 1.00 equiv) at 25°C under N2. The suspension was degassed and purged with H2 (50 Psi) three times. The mixture was heated to 45°C and stirred at 45°C under H2 (50 Psi) for 16 h. TLC (petroleum ether: ethyl acetate = 1:1) showed compound 3 (R f =0.60) is consumed, and a new spot is detected (R f =0.30). Three batches of the reaction were filtered and concentrated in vacuo, leaving only the DMA that would not come off the rotary evaporator. This was quenched in H2O (1.00 L) and filtered, and the filter cake was concentrated to give compound 4 (336 g, 1.50 mol, 97.5% yield, 99.5% purity) as an off-white solid.

[0092] LCMS: product: RT = 0.260 min, m / z = 224.1 (M+H) +

[0093] HPLC: Product: RT = 0.585 min, 99.5% purity under 220 nm.

[0094] SFC: (racemate), product: RT = 0.926 min, product: Rt = 1.507 mins

[0095] 1H NMR: 400MHz, DMSOδ / ppm: 7.13(d,J=8.8Hz,2H),6.53(d,J=8.4Hz,2H),5.29(s,2H),4.31-4.26(m,1H),4.17-4.11 (m,1H),3.08(dd,J=3.6,3.6Hz,1H),2.83-2.77(m,1H),2.76-2.68(m,1H),2.38-2.30(m,1H),2.06-1.96(m,1H).

[0096] Example 2 - Separation of enantiomers of compound 4

[0097] The two enantiomers of compound 4 were separated using supercritical fluid chromatography (SFC) with a chiral column.

[0098]

[0099] SFC column conditions:

[0100]

[0101] Compound 4 (332 g, 1.48 mol, 99.5% purity) was purified by preparative SFC (column: DAICEL CHIRALPAK AD (250 mm * 50 mm, 10 um); mobile phase: [Neu-MeOH]; B%: 50%-50%, 5.6; 2880 mins) to obtain R-4 (150 g, 666 mmol, 45.0% yield, 99.1% purity) and S-4 (162 g, crude product) in the form of a yellow solid. The crude product S-4 was ground with petroleum ether: ethyl acetate = 3: 1 (1.62 L) at 25 ° C for 2 h to obtain S-4 (132 g, 591 mmol, 40.0% yield, 99.8% purity) as a light yellow solid.

[0102] R-4

[0103] LCMS: product: RT = 0.663 min, m / z = 224.0 (M + H) +

[0104] HPLC: Product: RT = 1.419 mins, 99.1% purity at 220 nm.

[0105] SFC: (homochiral), product: RT = 0.912min, ee%: 99.4%

[0106] 1HNMR: 400MHz, DMSOδ / ppm: 7.13(d,J=8.8Hz,2H),6.53(d,J=8.4Hz,2H),5.29(s,2H),4.31-4.26(m,1H),4.17-4.11 (m,1H),3.08(dd,J=3.6,3.6Hz,1H),2.83-2.77(m,1H),2.76-2.68(m,1H),2.38-2.30(m,1H),2.06-1.96(m,1H)S-4

[0107] LCMS: product: RT = 0.656 min, m / z = 224.0 (M+H) +

[0108] HPLC: Product: RT = 1.358 mins, 99.8% purity at 220 nm.

[0109] SFC: (homochiral), product: RT = 1.491mins, ee%: 99.7%

[0110] 1 H NMR: 400MHz, DMSOδ / ppm: 7.13 (d, J = 8.4Hz, 2H), 6.53 (d, J = 8.8Hz, 2H), 5.29 (s, 2H), 4.31-4.26 (m, 1H), 4.17-4.1 1(m,1H),3.08(dd,J=3.6,3.6Hz,1H),2.83-2.77(m,1H),2.75-2.68(m,1H),2.38-2.30(m,1H),2.06-1.96(m,1H)

[0111] Example 3 - Conversion of R-4 to RA

[0112] Preparation of Compound R-6

[0113]

[0114] Compound R-4 (69.7 g, 309 mmol, 99.1% purity, 1.05 equiv) and DMAP (dimethylaminopyridine; 7.19 g, 58.9 mmol, 0.200 equiv), TEA (89.4 g, 883 mmol, 123 mL, 3.00 equiv) were added to a solution of compound 5 (Boland et al., Bioorg. Med. Chem. Lett. 2013, 6442-6446) (141 g, 294 mmol, 97.4% purity, 1.00 equiv) in DCM (1.41 L) at 20° C. under N2. T3P (propylphosphonic anhydride; 281 g, 442 mmol, 263 mL, 50.0% purity, 1.50 equiv) was added to the mixture at 20° C., and the mixture was stirred at 20° C. for 3 h. LCMS shows that compound 5 is consumed, and the required MS (RT = 0.922min) is detected. The reactant is diluted with DCM (1.40L) and washed with saturated NaHCO3 solution (1.40L*3). The organic layer is dried over Na2SO4, filtered and concentrated under reduced pressure to obtain a residue. The residue is purified by column chromatography (SiO2, petroleum ether: ethyl acetate = 1:0 to 0:1, petroleum ether: ethyl acetate = 0:1, R f =0.50) to obtain compound R-6 (195 g, 289 mmol, 98.0% yield, 99.3% purity) as a light yellow solid.

[0115] LCMS: product: RT = 0.922 min, m / z = 671.3 (M+H) +

[0116] SFC: (homochiral), product: RT = 4.269 mins, ee%: 100%

[0117] 1H NMR: 400MHz, DMSOδ / ppm: 10.5 (s, 1H), 10.37 (s, 1H), 8.59 (d, J = 2.4Hz, 1H), 8.39 (d, J = 5.6Hz, 1H) ,8.02-8.01(m,3H),7.93-7.90(m,2H),7.79(d,J=8.4Hz,2H),7.70-7.64(m,2H),7.57(d,J=8.0H z,1H),7.51(t,J=6.0Hz,1H),7.42(d,J=8.8Hz,2H),4.33-4.28(m,1H),4.19-4.12(m,3H),3.08- 3.02(m,1H),2.93-2.85(m,1H),2.38-2.31(m,1H),2.09-2.01(m,1H),1.37(s,9H),1.22(s,1H).

[0118] Preparation of RA

[0119]

[0120] At 35 ° C, HCl / dioxane (4.00M, 361mL, 10.0 equivalents) was added dropwise to a solution of compound R-6 (97.5g, 144mmol, 99.3% purity, 1.00 equivalents) in DCM (dichloromethane; 975mL) and dioxane (975mL). The mixture was stirred at 35 ° C for 2h. LCMS showed that compound R-6 was consumed and the required MS (RT = 0.773min) was detected. The reaction mixture was cooled to 25 ° C and stirred at 25 ° C for 12h. The mixture was concentrated under reduced pressure to obtain a residue. The residue was diluted with deionized H2O (2.00L), and the pH was adjusted to 7 with saturated NaHCO3 solution (300mL), and the filter cake was filtered and concentrated. The crude products from both reactions were combined, triturated with deionized H2O (1.50 L) at 25°C for 12 h, then filtered and the filter cake concentrated to afford RA HCl (110 g, 180 mmol, 62.2% yield, 99.1% purity, HCl) as a light yellow solid.

[0121] LCMS: product: RT = 0.773 min, m / z = 571.3 (M+H) +

[0122] HPLC: Product: RT = 7.942 mins, 99.1% purity at 220 nm.

[0123] Chiral NP-HPLC, product: Rt = 13.573mins, ee%: 100%

[0124] Chiral NP-HPLC conditions:

[0125]

[0126] EA-CHNS: C: 54.0%, H: 5.36%, N: 7.98%, S: 4.59%

[0127] 1 H NMR: 400MHz, DMSO δ / ppm: 10.7 (s, 1H), 10.56 (s, 1H), 8.60 (d, J=2.8Hz, 1H), 5.54 (s, 2H), 8.40 (d, J= 5.2Hz,1H),8.18(s,1H),8.11(dd,J=2.0,2.0Hz,1H),8.07-8.05(m,2H),7.91-7.88(m,2H),7.85(d ,J=8.8Hz,2H),7.72-7.67(m,2H),7.41(d,J=8.8Hz,2H),4.33-4.28(m,1H),4.18-4.12(m,1H),4.0 6(s,2H),3.32(s,1H),3.08-3.02(m,1H),2.91-2.87(m,1H),2.38-2.31(m,1H),2.09-2.02(m,1H).

[0128] Example 4 - Conversion of S-4 to SA

[0129] Preparation of compound S-6

[0130]

[0131] Compound S-4 (69.2 g, 309 mmol, 99.8% purity, 1.05 eq.) and DMAP (7.19 g, 58.9 mmol, 0.200 eq.), TEA (89.4 g, 883 mmol, 123 mL, 3.00 eq.) were added to a solution of compound 5 (Boland et al., Bioorg. Med. Chem. Lett. 2013, 6442-6446) (141 g, 294 mmol, 97.4% purity, 1.00 eq.) in DCM (1.41 L) at 20°C under N2. T3P (281 g, 442 mmol, 263 mL, 50.0% purity, 1.50 eq.) was added to the mixture at 20°C, and the mixture was stirred at 20°C for 3 h. LCMS showed that compound 5 was consumed, and the desired MS (RT = 0.917 min) was detected. The reactant was diluted with DCM (1.40 L) and washed with saturated NaHCO solution (1.40 L*3). The organic layer was dried over NaSO, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO, petroleum ether: ethyl acetate = 1:0 to 0:1, petroleum ether: ethyl acetate = 0:1, R f =0.50) to obtain compound S-6 (197 g, 290 mmol, 98.5% yield, 98.9% purity) as a light yellow solid.

[0132] LCMS: product: RT = 0.917 min, m / z = 671.3 (M+H) +

[0133] HPLC: Product: RT = 2.119 mins, 98.9% purity at 220 nm.

[0134] SFC: (homochiral), product: Rt = 5.810mins, ee%: 100%

[0135] 1H NMR: 400MHz, DMSOδ / ppm: 10.5 (s, 1H), 10.4 (s, 1H), 8.59 (d, J = 2.4Hz, 1H), 8.39 (d, J = 5.6Hz, 1H) ,8.02-8.01(m,3H),7.93-7.90(m,2H),7.79(d,J=8.8Hz,2H),7.70-7.64(m,2H),7.58(d,J=8.0H z,1H),7.51(t,J=6.0Hz,1H),7.42(d,J=8.4Hz,2H),4.33-4.28(m,1H),4.19-4.12(m,3H),3.08- 3.02(m,1H),2.93-2.85(m,1H),2.38-2.32(m,1H),2.09-2.01(m,1H),1.38(s,9H),1.22(s,1H).

[0136] Preparation of SA

[0137] At 35 ° C, HCl / dioxane (4M, 363 mL, 10.0 eq.) was added dropwise to a solution of compound S-6 (98.3 g, 145 mmol, 98.9% purity, 1.00 eq.) in DCM (983 mL) and dioxane (983 mL). The mixture was stirred at 35 ° C for 2 h. LCMS showed that compound S-6 was consumed and the required MS (RT = 0.774 min) was detected. The two batches of reaction mixture were cooled to 25 ° C and stirred at 25 ° C for 12 h. The mixture was concentrated under reduced pressure to obtain a residue. The residue was diluted with deionized H2O (2.00 L), and the pH was adjusted to 7 with saturated NaHCO3 solution (310 mL), and the filter cake was filtered and concentrated. The crude products from both reactions were combined, triturated with deionized H2O (1.50 L) at 25°C for 12 h, then filtered and the filter cake concentrated to afford S-AHCl (112 g, 182 mmol, 62.8% yield, 98.8% purity, HCl) as a light yellow solid.

[0138] LCMS: product: RT = 0.774 min, m / z = 571.3 (M+H) +

[0139] Chiral NP-HPLC, product: Rt = 11.407mins, ee%: 100%

[0140] Chiral NP-HPLC was the same as described in Example 3

[0141] EA-CHNS: C: 56.7%, H: 5.39%, N: 8.35%, S: 4.72%

[0142] 1 H NMR: 400MHz, DMSOδ / ppm: 10.7 (s, 1H), 10.5 (s, 1H), 8.60 (d, J = 2.8Hz, 1H), 8.41-8.40 (m, 3H), 8 .16(s,1H),8.11(dd,J=2.0,1.6Hz,1H),8.08-8.06(m,2H),7.91-7.87(m,2H),7.85(d,J=8.8H z,2H),7.72-7.67(m,2H),7.41(d,J=8.8Hz,2H),4.33-4.28(m,1H),4.18-4.12(m,1H),4.06(s ,2H),3.36(s,1H),3.08-3.02(m,1H),2.91-2.87(m,1H),2.37-2.31(m,1H),2.07-2.02(m,1H).

[0143] This method directly forms SA HCl Salt Form I. XRPD was performed on SA HCl Salt Form I using the following conditions:

[0144] XRPD conditions

[0145] XRPD diffractograms were collected using an X-ray diffractometer. The samples were prepared on zero-background silicon wafers by gently pressing onto a flat surface. The parameters for XRPD diffraction are shown below.

[0146] XRPD Testing Parameters

[0147]

[0148] XRPD of SA HCl Salt Form I (see also Figure 1 )

[0149]

[0150]

[0151] Example 5 - Formation of SA Succinate Form I

[0152] Formation of SA free base form I: At room temperature (24 ° C), about 100 mg of SA HCl salt form I was suspended in 1 mL of acetonitrile. After stirring for 15 min, a thick suspension was observed, and then 6 equivalents of NaHCO3 aqueous solution (84 mg NaHCO3 dissolved in 0.8 mL of water) were added. After stirring for 35 minutes, a suspension was observed. 2 mL of acetonitrile and 10 mL of water were added, and then after stirring for 10 min, a solid mixed with oil was precipitated. Finally, a suspension was obtained after stirring at room temperature for 15 h. The solid was collected by filtration and dried in vacuo at 40 ° C for about 3 h to obtain SA free base form I.

[0153] Formation of SA succinate form I: About 400 mg of SA free base form I and 1.1 equivalents of succinic acid (91 mg) were added to 8 mL of MEK (methyl ethyl ketone) at room temperature (about 24° C.). The mixture was stirred at room temperature for 4 h, and then the solid was collected by filtration and washed with MEK and dried at 40° C. under vacuum for 24 h to provide SA succinate form I.

[0154] XRPD was performed on SA succinate salt Form I. The conditions were the same as those described in Example 4 for the XRPD analysis of HCl salt Form I.

[0155] XRPD peaks of succinate form I (see also Figure 2 )

[0156]

[0157]

[0158] Example 6 - Formation of SA HCl Salt Form II

[0159] About 600 mg of SA HCl salt Form I was suspended in 20 mL of acetone / water (19 / 1, v / v) at room temperature and stirred for about 16 h. The solid was collected by filtration and dried under vacuum at 40° C. for about 2 h. About 545 mg of HCl salt Form II was prepared with a yield of 91%.

[0160] XRPD was performed on SA HCl Salt Form II. The conditions were the same as those described in Example 4 for the XRPD analysis of HCl Salt Form I.

[0161] XRPD peaks of SA HCl salt Form II (see also Figure 3 )

[0162]

[0163]

[0164] Example 7 - Stability Test Data

[0165] method:

[0166] About 10 mg of free base form I, HCl salt form II, succinate form I and accepted HCl salt form I were placed at 60°C / closed and 40°C / 75% RH (open), respectively. Two replicate samples were prepared for each condition. On the 0th day and the 7th day, samples were analyzed by HPLC and XRPD to check purity and crystalline form, respectively.

[0167] The results are shown in the following table.

[0168]

[0169]

[0170] All three salt forms are more stable than the free base Compound A. Both HCl salt forms are more stable than the succinate form.

[0171] The long term stability of the HCl salt Form II was also tested under the following conditions:

[0172] ●25℃±2℃,60%RH±5%RH

[0173] ●40℃±2℃,75%RH±5%RH.

[0174] Duplicate samples were prepared for each condition. At day 0, one month and 3 months, samples were analyzed by HPLC and XRPD to check purity and crystalline form, respectively.

[0175] The results are shown in the following table.

[0176] Stability of HCl salt form II at 25°C ± 2°C, 60% RH ± 5% RH

[0177]

[0178] Stability of HCl salt form II at 40°C ± 2°C, 75% RH ± 5% RH

[0179]

[0180] It can be seen that the HCl salt Form II is stable even under accelerated conditions for up to 3 months.

[0181] Example 8 - Solubility Test Data

[0182] method:

[0183] About 15 mg of sample was weighed into each sample bottle, and then 3.0 mL of water, simulated gastric fluid (SGF), fasting state simulated intestinal fluid (FaSSIF) or fed state simulated intestinal fluid (FeSSIF) medium was added. Triplicate samples were prepared for each medium. The suspension was kept shaking at 37 ° C for 24 h. At 0.5, 2 and 24 h, the suspension was filtered and the filtrate was analyzed by HPLC.

[0184] The results are shown in the following table.

[0185]

[0186]

[0187] HCl Salt Form II and Succinate Form I are more soluble in SGF than HCl Salt Form I.

[0188] Example 9 - Activity of R- and S-Compound A against Kinases Other Than ROCK

[0189] The HCl salt of the S and R enantiomers of Compound A were tested against a panel of kinases.

[0190] Compounds were received as powders and resuspended in 10 mM DMSO stock. Compounds were tested in a 10-dose IC50 format with 3-fold serial dilutions starting at 10 μM. The control compound, staurosporine, was tested in a 10-dose IC50 format with 4-fold serial dilutions starting at 20 μM. Reactions were performed at Km (Michaelis constant) ATP concentrations (as indicated in the table).

[0191] Reaction conditions: Buffer conditions: 20mM HEPES ((4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; pH 7.5), 10mMMgCl2, 1mM EGTA (ethylene glycol-bis(β-aminoethyl)-N,N,N′,N′-tetraacetic acid), 0.01% Brij35, 0.02mg / mlBSA (bovine serum albumin), 0.1mM Na3VO4, 2mM DTT (dithiothreitol), 1% DMSO. Reaction steps: 1. Prepare the specified substrate in freshly prepared reaction buffer. 2. Deliver the required cofactors to the above substrate solution. 3. Deliver the specified kinase to the substrate solution and mix gently. 4. Use acoustic technology (Echo550) to deliver the compounds in DMSO to the kinase reaction mixture. 5. 33P-ATP (final specific activity of 0.01 μCi / μl) was delivered to the reaction mixture to initiate the reaction. 6. Incubate the kinase reaction at room temperature for 120 min. 7. Spot the reaction on P81 ion exchange paper (Whatman #3698-915). 8. Wash the filter thoroughly with 0.75% phosphoric acid. 9. Measure the radioactive phosphorylated substrate remaining on the filter paper. Data Analysis: Kinase activity data are expressed as the percentage of kinase activity remaining in the test sample compared to the vehicle (dimethyl sulfoxide) reaction. IC50 values ​​and curve fitting were obtained using Prism4 software (GraphPad).

[0192] The results are shown in the following table.

[0193]

[0194] For 6 of the 7 off-target kinases, the S-enantiomer of compound A was less active than the R-enantiomer. For only 1 of the 7 off-target kinases, the R-enantiomer of compound A was less active than the S-enantiomer. This is particularly surprising because to date, the inventors have not detected a statistically significant difference between the activity of the S-enantiomer and the R-enantiomer of compound A against the kinases ROCK1 or ROCK2.

[0195] Example 10 - Activity of R- and S-Compound A on CEREP Panel

[0196] In vitro pharmacology screens against a variety of targets (receptors, ion channels, enzymes, and transporters) are used to identify undesirable off-target activities that may lead to adverse drug reactions in humans. Screening panels (such as SafetyScreen44 TM The S and R enantiomers of Compound A were tested against a panel of targets that included a series of off-targets that were associated with known problems in humans and that, if discovered after a drug was approved, could impede or halt the development of a candidate drug or cause it to be withdrawn from the market.

[0197] Typical adverse drug reactions associated with these off-targets are summarized (Bowes et al., Nature Reviews Drug Discovery 2012, 11, 909).

[0198] G protein coupled receptors:

[0199] Cannabinoid receptor CB1

[0200] Stimulant / Activating Effects: Euphoria and dysphoria; anxiety; memory impairment and difficulty concentrating; analgesia; hypothermia

[0201] Antagonistic effects: Increased weight loss; vomiting; depression

[0202] Cannabinoid receptor CB2

[0203] Antagonistic effects: Increased inflammation; decreased bone mass

[0204] Muscarinic acetylcholine receptor M1

[0205] Agonists: Convulsions; increased gastric acid secretion; hypertension; tachycardia; hyperthermia

[0206] Antagonists: Decreased cognitive function; decreased gastric acid secretion; blurred vision

[0207] Muscarinic acetylcholine receptor M2

[0208] Agonists: Decreased heart rate; reflexes; increased blood pressure; negative chronotropy and inotropy; decreased cardiac conduction; decreased duration of cardiac action potential

[0209] Antagonists: Tachycardia; bronchoconstriction; fibrillation

[0210] μ-opioid receptor (mu-MOP)

[0211] Agonists: Sedation; decreased gastrointestinal motility; pupil constriction; abuse liability; respiratory depression; miosis; hypothermia

[0212] Antagonists: Increased gastrointestinal motility; dyspepsia; flatulence

[0213] Enzymes

[0214] Monoamine oxidase A (MAO)

[0215] Inhibitory effects: Increased blood pressure when combined with amines such as tyramine; potential for drug interactions; dizziness; sleep disturbances; nausea

[0216] Transporter

[0217] Norepinephrine transporter uptake inhibition: Increased heart rate; increased blood pressure; increased motor activity; constipation; abuse potential

[0218] Serotonin transporter uptake inhibition: Increased gastrointestinal motility; decreased upper gastrointestinal transit; decreased plasma renin; increased other serotonin-mediated effects; insomnia; anxiety; nausea; sexual dysfunction

[0219] The tests were performed according to the methods described in the table below and in the references indicated.

[0220] CEREP Group:

[0221]

[0222]

[0223]

[0224]

[0225]

[0226] 1. Cesura, AM et al., (1990), Mol. Pharmacol., 37: 358-366.

[0227] 2. Felder, CC et al., (1995), Mol. Pharmacol., 48: 443-450.

[0228] 3. Sur, C. et al., (2003), Proc. Natl. Acad. Sci. USA, 100: 13674-13679.

[0229] 4. Michal, P. et al., (2001), Brit. J. Pharmacol., 132: 1217-1228.

[0230] 5. Wang, JB et al., (1994), FEBS Lett., 338: 217-222.

[0231] 6. Perovic, S. and Muller, WEG (1995), Arzneim-Forsch. Drug Res., 45: 1145-1148.

[0232] 7. Verrico C. et al., (2007), Psychopharmacology, 189, 489-503.

[0233] The results are shown in the following table.

[0234]

[0235]

[0236] In seven out of thirteen assays, the S-enantiomer of Compound A was less active than the R-enantiomer. In only one out of thirteen assays was the R-enantiomer of Compound A less active than the S-enantiomer.

[0237] The combination of this data for the CEREP panel and the data presented in Example 9 for the kinase panel suggests that the S-enantiomer is more selective than the R-enantiomer for multiple off-target proteins and is less likely to be associated with undesirable toxicities and side effects that limit efficacy and tolerability in humans.

Claims

1. S-enantiomer (SA) of compound A: It is in substantially enantiomerically pure form, or a pharmaceutically acceptable salt thereof.

2. Compound A according to claim 1, wherein the compound A is in the form of a free base.

3. Compound A according to claim 1, wherein the compound A is in the form of a pharmaceutically acceptable salt.

4. Crystalline salt form of compound A:

5. The crystalline salt form of claim 4, wherein the salt is the hydrochloride salt.

6. The crystalline salt form of claim 4, wherein the salt is succinate.

7. The crystalline salt form of any one of claims 4 to 6, wherein Compound A is racemic.

8. The crystalline salt form according to any one of claims 4 to 6, wherein Compound A is substantially in the form of a single enantiomer.

9. The crystalline salt form of claim 8, wherein the enantiomer is the S-enantiomer (SA):

10. The crystalline salt form of claim 8, wherein the enantiomer is the R-enantiomer (RA):

11. The crystalline salt form of claim 5, wherein the salt is the hydrochloride salt of a single enantiomer of compound A, characterized in that When using K α2 / K α1 The crystalline salt form has an XRPD pattern having at least two peaks at 2θ selected from 16.2±0.2, 22.7±0.2, 23.3±0.2, 24.0±0.2, 24.9±0.2 and 25.4±0.2 when measured using Cu radiation at a ratio of 0.

5.

12. The crystalline salt form according to claim 11, characterized in that When using K α2 / K α1 The crystalline salt form has an XRPD pattern having peaks at 2θ selected from 16.2±0.2, 22.7±0.2, 23.3±0.2, 24.0±0.2, 24.9±0.2 and 25.4±0.2 when measured with Cu radiation at a ratio of 0.

5.

13. The crystalline salt form according to claim 11, characterized in that When using K α2 / K α1 The crystalline form has an XRPD pattern substantially as shown in FIG3 when measured with Cu radiation at a ratio of 0.

5.

14. The crystalline salt form of claim 5, wherein the salt is the hydrochloride salt of a single enantiomer of Compound A, characterized in that When using K α2 / K α1 The crystalline salt form has an XRPD pattern having at least two peaks at 2θ selected from 13.6±0.2, 14.4±0.2, 14.5±0.2, 16.2±0.2 and 16.5±0.2 when measured using Cu radiation at a ratio of 0.

5.

15. The crystalline salt form according to claim 14, characterized in that When using K α2 / K α1 The crystalline salt form has an XRPD pattern having peaks at 2θ selected from 13.6±0.2, 14.4±0.2, 14.5±0.2, 16.2±0.2 and 16.5±0.2 when measured using Cu radiation at a ratio of 0.

5.

16. The crystalline salt form of claim 14, wherein When using K α2 / K α1 The crystalline form has an XRPD pattern substantially as shown in Figure 1 when measured with Cu radiation at a ratio of 0.

5.

17. The crystalline salt form of claim 6, wherein the salt is a succinate salt of a single enantiomer of Compound A, characterized in that When using K α2 / K α1 The crystalline salt form has an XRPD pattern having at least two peaks at 2θ selected from 18.2±0.2, 18.6±0.2, 19.1±0.2, 21.4±0.2, 23.0±0.2, 24.1±0.2 and 25.8±0.2 when measured with Cu radiation at a ratio of 0.

5.

18. The crystalline salt form according to claim 17, characterized in that When using K α2 / K α1 The crystalline salt form has an XRPD pattern having peaks at 2θ selected from 18.2±0.2, 18.6±0.2, 19.1±0.2, 21.4±0.2, 23.0±0.2, 24.1±0.2 and 25.8±0.2 when measured using Cu radiation at a ratio of 0.

5.

19. The crystalline salt form of claim 17, wherein When using K α2 / K α1 The crystalline form has an XRPD pattern substantially as shown in FIG2 when measured with Cu radiation at a ratio of 0.

5.

20. A pharmaceutical composition comprising Compound A according to any one of claims 1 to 3 or Compound A in the form of a crystalline salt according to any one of claims 4 to 19.

21. Compound A according to any one of claims 1 to 3 or a crystalline form according to any one of claims 4 to 19 for use in the treatment of a fibrotic disease.

22. The salt form of claim 21, wherein the fibrotic disease is a disease of the gastrointestinal system.

23. The salt form of claim 21, wherein the fibrotic disease is fibrostenotic Crohn's disease.

24. A method for preparing a single enantiomer of Compound A, HCl crystalline salt Form II, the method comprising: a) suspending a sample of the HCl crystalline salt Form I of a single enantiomer of Compound A in a solvent to form a suspension; b) filtering the suspension to obtain HCl crystalline salt Form II.

25. The method of claim 24, wherein the solvent is a mixture of acetone and water.

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