Salt of omecamtiv mecarbil and process for preparing salt

Abstract

Dihydrochloride hydrated salt of omecamtiv mecarbil. The application contains 14 more patent claims.

Description

Description AREA

[0001] Provided are dihydrochloride hydrate salts of omecamtiv mecarbil and polymorphic forms thereof and methods for preparing dihydrochloride hydrate salts of omecamtiv mecarbil, including polymorphic forms of omecamtiv mecarbil dihydrochloride hydrate. Also disclosed are compositions containing polymorphic forms of omecamtiv mecarbil dihydrochloride and methods of using polymorphic forms of omecamtiv mecarbil dihydrochloride. BACKGROUND

[0002] Cardiac sarcomeres are the basic unit of muscle contraction in the heart. Cardiac sarcomeres are highly ordered cytoskeletal structures composed of myosin, actin, and a set of cardiac muscle regulatory proteins. The discovery and development of small molecule cardiac myosin activators would lead to promising treatments for acute and chronic heart failure. Cardiac myosin is the master of cytoskeletal motors in the cardiac muscle cell. It is directly responsible for converting chemical energy into mechanical force, resulting in cardiac muscle contraction.

[0003] Current positive inotropic agents, such as beta-adrenergic receptor agonists or phosphodiesterase inhibitors, increase intracellular calcium concentration and thereby increase cardiac sarcomere contractility. However, the increase in calcium levels increases the rate of cardiac muscle contraction and shortens the systolic ejection time, which is associated with potentially life-threatening adverse effects. In contrast, cardiac myosin activators act by a mechanism that directly stimulates the activity of the cardiac muscle motor protein myosin without increasing intracellular calcium concentration. They accelerate the rate-limiting step of the myosin enzymatic cycle and shift it to a force-generating state. Rather than increasing cardiac contraction rate, this mechanism instead prolongs systolic ejection time, resulting in increased cardiac muscle contractility and cardiac output in a potentially more oxygen-efficient manner.

[0004] U.S. Patent No. 7,507,735 discloses a class of compounds, including an emollient mecarbil (AMG 423, CK-1827452), which have the structure:

[0005] US 2006 / 014761 A1 relates to "certain substituted urea derivatives" which allegedly "selectively modulate the cardiac sarcomere".

[0006] Omecamtiv mecarbil is a first-in-class direct activator of cardiac myosin, the motor protein that drives cardiac contraction. It is being evaluated as a potential treatment for heart failure in intravenous and oral formulations with the goal of establishing a new continuum of care for patients in both inpatient and outpatient settings.

[0007] As drug compounds having, for example, improved stability, solubility, shelf life, and in vivo pharmacology are continually sought, there is a continuing need for new or purer salts, hydrates, solvates, and polymorphic crystalline forms of existing drug molecules. The crystalline forms of omecamtiv mecarbil described herein help to meet these and other needs. SUMMARY

[0008] The scope of the invention is defined by the claims.

[0009] The dihydrochloride hydrate salt of the emollient mecarbyl is provided.

[0010] Also provided is omecamtiv mecarbil dihydrochloride hydrate.

[0011] Also provided is a crystalline form of the dihydrochloride hydrate salt of omecamtiv mecarbil.

[0012] Also provided is omecamtiv mecarbil dihydrochloride hydrate Form A.

[0013] Also disclosed herein are compositions and pharmaceutical compositions comprising the dihydrochloride form of omecamtiv mecarbil.

[0014] Also provided is a process for preparing omecamtiv mecarbil dihydrochloride hydrate. salt consisting of mixing methyl 4-(3-amino-2-fluorobenzyl)piperazine-1-carboxylate and phenyl(6-methylpyridin-3-yl)carbamate in the presence of a trialkylamine base to form omecamtiv mecarbil; and further comprising crystallizing omecamtiv mecarbil in the presence of aqueous hydrochloric acid and an alcoholic solvent to form omecamtiv mecarbil dihydrochloride hydrate salt, for example, wherein the alcoholic solvent comprises isopropyl alcohol.

[0015] Also provided is a process for preparing omecamtiv mecarbil dihydrochloride hydrate comprising: (a) hydrogenating methyl 4-(2-fluoro-3-nitrobenzyl)piperazine-1-carboxylate in the presence of a hydrogenation catalyst to give methyl 4-(3-amino-2-fluorobenzylpiperazine-1-carboxylate); (b) mixing methyl 4-(3-amino-2-fluorobenzyl)piperazine-1-carboxylate and phenyl(6-methylpyridin-3-yl)carbamate in the presence of a trialkylamine base to form the omecamtiv mecarbil as the free base; and (c) crystallization of omecamtiv mecarbil free base in the presence of aqueous hydrochloric acid and an alcoholic solvent to form omecamtiv mecarbil dihydrochloride hydrate salt. DESCRIPTION OF IMAGES

[0016] Figure 1 shows the dynamic vapor sorption of omecamtiv mecarbil dihydrochloride hydrate form, Form A. Figure 2 shows the X-ray powder diffraction (XRPD) pattern of Form A. Figure 3 shows the XRPD patterns of the salt formula of omecamtiv mecarbil dihydrochloride hydrate under different relative humidity conditions. Figure 4 shows the XRPD form of the mecamtiv mecarbyl dihydrochloride hydrate salt form at different temperatures. Figure 5 shows a thermograph of differential scanning calorimetric and thermogravimetric analysis of Form A. Figure 6 shows an overlay of XRPD patterns for Forms A, B, and C of the omecamtiv mecarbyl dihydrochloride salt. Figure 7 shows the drug release at two pHs (2 and 6.8) for the free base formulation of omecamtiv mecarbil (top) and for the salt form of omecamtiv mecarbil dihydrochloride. hydrate, Form A (bottom). DETAILED DESCRIPTION

[0017] Unless otherwise specified, the following definitions apply to terms found in the specification and claims: "Treatment" or "treat" means any treatment of a disease in a patient, including: a) preventing the disease, i.e. causing the clinical symptoms of the disease not to develop; b) inhibiting the disease; c) slowing or stopping the development of clinical symptoms; and / or d) alleviating the disease, i.e. causing regression of clinical symptoms. Treatment of diseases and disorders herein is intended to encompass the prophylactic use of a pharmaceutical formulation described herein to a subject (i.e., an animal, preferably a mammal, most preferably a human) believed to be in need of preventive treatment, such as, for example, chronic heart failure.

[0018] The term "therapeutically effective amount" refers to an amount that is effective, when administered to a human or non-human subject, to treat a disease, e.g., a therapeutically effective amount may be an amount sufficient to treat a disease or disorder responsive to myosin activation. A therapeutically effective amount may be determined experimentally, for example by determining the concentration of the chemical entity in blood, or theoretically, by calculating bioavailability.

[0019] "Pharmaceutically acceptable salts" include, but are not limited to salts with inorganic acids, such as hydrochlorides (ie, hydrochlorides), phosphates, diphosphates, hydrobromates, sulfates, sulfinates, nitrates, and similar salts; as well as organic acid salts such as malate, maleate, fumarate, tartrate, succinate, citrate, acetate, lactate, methanesulfonate, p-toluenesulfonate, 2-hydroxyethylsulfonate, benzoate, salicylate, stearate and alkanoate such as acetate, HOOC-(CH2) n-COOH where n is 0-4, and similar salts. Similarly, pharmaceutically acceptable cations include, but are not limited to, sodium, potassium, calcium, aluminum, lithium, and ammonium. Those skilled in the art will recognize various synthetic methodologies that can be used to prepare non-toxic pharmaceutically acceptable addition salts.

[0020] As used herein, the term "polymorphic" or "polymorphic forms" refers to crystalline forms of the same molecule. Different polymorphic forms of molecules have different physical properties as a result of the arrangement or conformation of molecules in the crystal lattice. Some of the different physical properties include melting point, heat of fusion, solubility, velocity dissolution and / or vibrational spectra. The physical form of a particular compound is particularly important when the compound is used in a pharmaceutical formulation, as different solid forms of the compound result in different drug properties.

[0021] Polymorphs of molecules can be obtained by a number of methods, as shown in the art, such as, for example, melt recrystallization, melt cooling, solvent recrystallization, decomposition, rapid evaporation, rapid cooling, slow cooling, vapor diffusion, and sublimation. Techniques for characterizing polymorphs include X-ray diffraction pattern analysis (XRPD), single crystal X-ray diffraction (XRD), differential scanning calorimetry (DSC), vibrational spectroscopy (e.g., IR and Raman spectroscopy), solid-state nuclear magnetic resonance (ssNMR), hot-stage optical microscopy, scanning electron microscopy (SEM), electron crystallography and quantitative analysis, particle size analysis (PSA), surface area analysis, solubility studies, and dissolution studies.

[0022] The term "hydrate" refers to a chemical entity formed by the interaction of water and a compound.

[0023] As used herein, the term "monohydrate" refers to a hydrate containing one molecule of water per molecule of substrate.

[0024] As used herein, the term "crystalline" refers to a solid in which the constituent atoms, molecules, or ions are arranged in a regularly ordered, repeating pattern in three dimensions.

[0025] The specification and claims contain a list of species using the language "selected from ... and ..." and "is ... or ..." (sometimes referred to as Marcus groups). When this language is used in this application, unless otherwise indicated, it is intended to include the group as a whole or any member thereof or any subgroup thereof. The use of this language is for brevity only and is not in any way limited to the elimination of individual elements or subgroups.

[0026] A dihydrochloride hydrate salt form of omecamtiv mecarbil is provided. In various embodiments of the present aspect, the dihydrochloride hydrate form of omecamtiv mecarbil is crystalline (Form A). Embodiments of the dihydrochloride hydrate form of omecamtiv mecarbil can be specified by one or more parameters which are described in more detail below.

[0027] The dihydrochloride hydrate form of omecamtiv mecarbil has a water solubility of greater than 40 mg / mL at a pH in the range of about 3.5. Further, Form A is not hygroscopic. For example, when subjected to dynamic vapor sorption, Form A exhibited an overall weight gain of about 0.55% by weight between about 40% and about 95% relative humidity (RH) and a weight loss of about 2.7% between about 30% and about 5% RH. In some embodiments, the dihydrochloride hydrate form of omecamtiv mecarbil has a dynamic vapor sorption profile substantially as shown in Figure 1, where "substantially" is understood to mean that the reported DVS characteristics may vary by about ±5% RH.

[0028] Dynamic vapor sorption indicated that the salt dehydrates when dried to 5% relative humidity, but almost completely rehydrates by 15% relative humidity. Above 15% relative humidity, the sample is not hygroscopic, showing only about 1.0% weight change after reaching 95% relative humidity. No phase change occurred after the vapor sorption experiment when examined by XRPD.

[0029] It is soluble that the water solubility for Form A is greater than 40 mg / mL (pH = 3.5) without applying a phase change during the 24-hour suspension experiment when examined by XRPD. Furthermore, Form A is stable under conditions of accelerated stability testing. For example, Form A remains in the same physical form for 6 months at 40°C and 75% relative humidity.

[0030] In various embodiments, Form A may be characterized by X-ray powder diffraction, obtained as set forth in the examples, having peaks around 6.6, 14.9, 20.1, 21.4 and 26.8 ± 0.2° 20 using Cu Kα radiation. Form A can optionally be further characterized by X-ray powder diffraction which has additional maxima at about 8.4, 24.2, 26.0, 33.3 ± 0.2° 20 using Cu Kα radiation. Form A can optionally be further characterized by X-ray powder diffraction with additional maxima at about 6.2, 9.7, 13.2, 14.3, 15.4, 16.3, 16.9, 18.9, 19.5, 20.7, 21.8, 22.8, 23.6, 25.1, 27.3, 27.7, 28.4, 29.4, 30.2, 31.2, 31.5, 31.9, 33.9, 34.5, 34.9, 36.1, 36.8, 37.7, 38.5, and 39.7± 0.2° 20 using Cu Kα radiation. In various cases, Form A can be characterized by an XRPD pattern having maxima at about 6.2, 6.6, 8.4, 9.7, 13.2, 14.3, 14.9, 15.4, 16.3, 16.9, 18.9, 19.5, 20.1, 20.7, 21.4, 21.8, 22.8, 23.6, 24.3, 25.1, 26.0, 26.8, 27.3, 27.7, 28.4, 29.4, 30.2, 31.2, 31.5, 31.9, 33.3, 33.9, 34.5, 34.9, 36.1, 36.8, 37.7, 38.5, and 39.7± 0.2° 20 using Cu Kα radiation. In some embodiments, Form A has an X-ray powder diffraction pattern, as shown in Figure 2, where "basically" is understood to mean that the reported maxima may vary by about ± 0.2°. It is well known in the XRPD field that while the relative peak heights in spectra are dependent on a number of factors, such as sample preparation and instrument geometry, the peak positions are relatively insensitive to experimental details.

[0031] The Form B and Form C polymorphs of omecamtiv mecarbil, are anhydrous dihydrochloride metastable forms and can be formed under different hydration conditions, as shown in Figures 3, 4 and 6. Characteristic values ​​of Form B 2-theta include 6.8, 8.8, 14.7, 17.7 and 22.3 ± 0.2° 20 using Cu Kα radiation, and may additionally include peaks at 9.6, 13.5, 19.2, 26.2 ± 0.2° 20 using Cu Kα radiation. Form B can be characterized by XRPD peaks of the sample at 6.2, 6.8, 8.8, 9.6, 13.5, 14.4, 14.7, 15.4, 16.3, 17.0, 17.7, 18.3, 19.2, 19.9, 20.5, 20.8, 21.8, 22.3, 22.7, 23.0, 24.8, 25.1, 25.5, 26.2, 26.4, 26.8, 27.5, 28.5, 30.2, 30.6, 31.1, 31.5, 32.1, 32.7, 34.1, 34.4, 35.5, 35.9, 38.1, 38.9± 0.2° 20 using Cu Kα radiation. Characteristic values ​​of the C 2-theta form include 6.7, 14.8, 17.4, 20.6, and 26.2± 0.2° 20 using Cu Kα radiation, and may additionally include peaks at 8.7, 22.0, 27.1, and 27.7 ± 0.2° 20 using Cu Kα radiation. Form C can be characterized by XRPD peaks of the sample at 6.2, 6.7, 8.7, 9.6, 13.5, 14.5, 14.8, 15.4, 16.4, 17.1, 17.4, 18.4, 19.3, 19.5, 19.9, 20.6, 20.8, 21.8, 22.0, 22.5, 22.8, 24.3, 24.7, 25.1, 25.6, 26.2, 26.5, 27.1, 27.3, 27.7, 28.5, 30.0, 30.5, 31.0, 31.5, 32.2, 32.8, 34.1, 35.2, 36.0, 36.9, and 38.8± 0.2° 20 using Cu Kα radiation. Forms B and C may have a powder X-ray diffraction pattern essentially as shown in Figure 6, where "essentially" means that the reported maxima may vary by about ± 0.2°.

[0032] In various embodiments, Form A can be characterized by a single crystal X-ray diffraction pattern, obtained as set forth in the Examples section, wherein Form A has a triclinic space group P-1 and unit cell parameters of about a = 5.9979(4) A, b = 13.4375(9) A, c = 14.4250(9) A, a = 97.617(4)°; θ = 93.285(4)°; and y = 94.585(5)°. Form A can optionally be further characterized by the XRD parameters in the table below. Wavelength 1.54178 A Crystal system Triclinic Space unit P-1 Dimensions of unit cell a = 5.9979(4) A a = 97.617(4)° b = 13.4375(9) AP = 93.285(4)° c = 14.4250(9) A 7 = 94.585(5)° Volume 1145.93(13) A 3 Z 2 Density (calculated) 1,427 Mg / m 3 Absorption coefficient 2.945 mm' 1

[0033] DSC thermograms were obtained for Form A. The DSC curve indicates an endothermic transition that appears to melt / decompose at about 235°C. Accordingly, in embodiments, Form A can be characterized by a DSC thermogram that has a decomposition endotherm with an onset in the range of about 230°C to about 240°C when Form A is in an open aluminum container. For example, in embodiments where Form A is heated to about 25°C at a rate of about 10°C / min, Form A can be characterized by a DSC thermogram that has a decomposition endotherm with an onset at about 235°C, as shown in Figure 5.

[0034] Form A can also be characterized by thermogravimetric analysis (TGA). Accordingly, Form A can be characterized by a weight loss in the range of about 2% to about 5% with an initial temperature in the range of about 100°C to about 150°C. For example, Form A can be characterized by a weight loss of about 3%, up to 150°C. In some embodiments, Form A has a thermogravimetric analysis substantially as shown in Figure 5, where "substantially" is understood to mean that the reported TGA characteristics may vary by about ± 5°C. For this weight loss, water was determined by Karl Fischer (KF) analysis. The KF analysis indicates that the water content of Form A can be about 3.7, which is consistent with the monohydrate.

[0035] Form A can be characterized by XRPD temperature variation and XRPD relative humidity variation. The XRPD temperature variation data are shown in Figure 4. The data show that when the hydrate of Form A is heated through a desolvation event, the shown on the TGA curve (around 75°C), the material transforms into a new dehydrated phase, Form B. When the material is cooled back to ambient conditions, Form B resorbs water from the atmosphere and transforms back into hydrated Form A. The XRPD data of variable relative humidity is shown in Figure 3. The data show that when hydrated Form A is exposed to 5% relative humidity, the material transforms into a new dehydrated phase, Form C. When the material is exposed to 15% relative humidity and higher, Form C resorbs water from the environment and transforms into hydrated Form A. These data are consistent with the vapor sorption experiment. The overlay of Form B and Form C is shown in Figure 6. The arrows mark significant reflections of the two powder samples, indicating that the two phases are unique.

[0036] Compositions are disclosed that comprise the dihydrochloride hydrate form of omecamtiv mecarbil. The compositions may comprise at least about 50, about 60, about 70, about 80, about 90, about 95, about 96, about 97, about 98, or about 99% by weight of the dihydrochloride hydrate form of omecamtiv mecarbil. The compositions may comprise at least about 50, about 60, about 70, about 80, about 90, about 95, about 96, about 97, about 98, or about 99% by weight of Form A of the dihydrochloride hydrate form of omecamtiv mecarbil. The compositions may comprise a mixture of two or more of Forms A, B, and C.

[0037] Pharmaceutical formulations are provided comprising the dihydrochloride hydrate form of omecamtiv mecarbil and at least one pharmaceutically acceptable excipient. The compositions may comprise at least about 50, about 60, about 70, about 80, about 90, about 95, about 96, about 97, about 98, or about 99% by weight of the dihydrochloride hydrate form of omecamtiv mecarbil. The compositions may comprise at least about 50, about 60, about 70, about 80, about 90, about 95, about 96, about 97, about 98, or about 99% by weight of Form A of the dihydrochloride hydrate form of omecamtiv mecarbil. The formulations may comprise a mixture of two or more of Forms A, B, and C.

[0038] Also disclosed is a method for using such pharmaceutical formulations for the treatment of heart failure, including, but not limited to: acute (or decompensated) congestive heart failure and chronic congestive heart failure; particularly diseases associated with systolic heart dysfunction.

[0039] Also provided is a synthesis of omecamtiv mecarbil dihydrochloride hydrate salt consisting of mixing methyl 4-(3-amino-2-fluorobenzyl)piperazine-1-carboxylate and phenyl(6-methylpyridin-3-ylcarbamate) in the presence of a trialkylamine base to form an omecamtive mecarbyl; and further comprising crystallizing omecamtiv mecarbil in the presence of aqueous hydrochloric acid and an alcoholic solvent to form omecamtiv mecarbil dihydrochloride hydrate salt, for example, wherein the alcoholic solvent comprises isopropyl alcohol.

[0040] In some embodiments, the weight ratio of phenyl (6-methylpyridin-3-yl)carbamate hydrochloride (i.e., SM-2 or phenyl carbamate) to methyl 4-(3-amino-2-fluorobenzyl)piperazine-1-carboxylate (i.e., SM-1 or piperazine nitro) is between about 1.1 and 1.5. In some embodiments, the weight ratio of phenyl (6-methylpyridin-3-yl)carbamate hydrochloride to methyl 4-(3-amino-2-fluorobenzyl)piperazine-1-carboxylate is about 1.2.

[0041] In some embodiments, the mixing is carried out in the presence of an aprotic solvent. In some embodiments, the solvent is THF.

[0042] In some embodiments, the trialkylamine base is triethylamine, diisopropylethylamine, or a combination thereof. In some embodiments, the trialkylamine base comprises diisopropylethylamine.

[0043] In some embodiments, an excess of trialkylamine base is used. In some embodiments, between about 1.1 and 1.5 equivalents of trialkylamine base are used. In some embodiments, about 1.3 equivalents of trialkylamine base are used.

[0044] In some embodiments, mixing is performed at 65 °C.

[0045] In some embodiments, the alcoholic solvent comprises isopropyl alcohol.

[0046] In some embodiments, the aqueous hydrochloric acid comprises 6N HCl.

[0047] In some embodiments, the method further includes mixing omecamtiv mecarbil dihydrochloride hydrate with at least a pharmaceutically acceptable excipient to form a pharmaceutical formulation.

[0048] In some embodiments, the pharmaceutical formulation comprises omecamtiv mecarbil dihydrochloride hydrate; a swelling layer; a semipermeable membrane coating having at least one delivery aperture. General characteristics of the drug layer and the swelling layer can be found in U.S. Patent No. obj. 2011 / 0182947.

[0049] In some embodiments, the pharmaceutical formulation is a modified-release tablet matrix comprising omecamtiv mecarbil dihydrochloride hydrate; a controlled-release agent; a pH modifying agent; a filler; and a lubricant.

[0050] In some embodiments, methyl 4-(3-amino-2-fluorobenzyl)piperazine-1-carboxylate is prepared by a process comprising: hydrogenating methyl 4-(2-fluoro-3-nitrobenzyl)piperazine-1-carboxylate in the presence of a hydrogenation catalyst to produce methyl 4-(3-amino-2-fluorobenzyl)piperazine-1-carboxylate.

[0051] In some embodiments, the hydrogenation catalyst comprises palladium. In some embodiments, the hydrogenation catalyst is palladium on carbon.

[0052] Also disclosed is a process for preparing omecamtiv mecarbil dihydrochloride hydrate comprising crystallizing omecamtiv mecarbil in the presence of aqueous hydrochloric acid and an alcoholic solvent to form omecamtiv mecarbil dihydrochloride hydrate.

[0053] The alcohol solvent may contain isopropyl alcohol.

[0054] Also provided is a process for preparing omecamtiv mecarbil dihydrochloride hydrate comprising: (a) hydrogenating methyl 4-(2-fluoro-3-nitrobenzyl)piperazine-1-carboxylate in the presence of a hydrogenation catalyst to give methyl 4-(3-amino-2-fluorobenzylpiperazine-1-carboxylate); (b) mixing methyl 4-(3-amino-2-fluorobenzyl)piperazine-1-carboxylate and phenyl(6-methylpyridin-3-yl)carbamate in the presence of a trialkylamine base to form the omecamtiv mecarbil as the free base; and (c) crystallization of omecamtiv mecarbil free base in the presence of aqueous hydrochloric acid and an alcoholic solvent to form omecamtiv mecarbil dihydrochloride hydrate salt.

[0055] This synthesis gives high overall yields (greater than 70%). In addition, the dihydrochloride salt, which results from the step, can form as long rods when crystallized, has improved bulk properties, filtration times of minutes (compared to days for the free base form) and are highly soluble (greater than 40 mg / mL at pH 3.8). In various cases, the salt obtained is the dihydrochloride hydrate Form A. EXAMPLES

[0056] The scope of the invention is defined by the claims. Any example that does not fall within the scope of the appended claims is provided for reference purposes. General procedures

[0057] Reagents and solvents were used as obtained from commercial sources. 'H NMR spectra were recorded on a 400 MHz spectrometer. Chemical shifts are reported in ppm from tetramethylsilane with solvent resonances as internal standards (CDCl3, DMSO-d6). Data are reported as follows: chemical shift, multiplicity (s = singlet, d = doublet, t = triplet, k = quartet, br = broad, m = multiplet), coupling constants (Hz) and integration. 13C NMR spectra were recorded on a 100 MHz spectrometer with total proton separation. Chemical shifts are reported in ppm from tetramethylsilane with solvent as internal reference (CDCl3, DMSO-de). All solvent charges are made relative to the starting 2-fluoro-3-nitrotoluene.

[0058] X-ray powder diffraction data were obtained using a Philips Automated X-ray Powder Diffraction (X'Pert) equipped with a fixed slit. The radiation was Cu Kα (1.541837A), and the voltage and current were 45 kV and 40 mA, respectively. Data were collected at room temperature from 3,000 to 40,009 degrees 2-theta; the step size was 0.008 degrees; the counting time was 15,240 seconds. Samples ranging from 5-40 mg were prepared on a sample holder and the stage was rotated with a rotation time of 2,000 seconds.

[0059] The thermal properties of omecamtiv mecarbil bis-HCl salt were characterized using a DSC Q1000 or DSC K 100 model, TA Instruments, differential scanning calorimetry and a Q 500, TA Instruments, thermogravimetric analyzer. Data analysis was performed using Universal Analysis 2000, TA Instruments. Heating rates of 10 °C / min were used at different temperature intervals for differential scanning calorimetry and thermogravimetric analysis. Samples ranging from <1-5 mg were prepared in sealed, sealed or open aluminum pans for DSC analysis.

[0060] Moisture balance data were collected using a VTI SGA 100 symmetric vapor sorption analyzer. The relative humidity was varied in 5% increments, between 5% and 95% relative humidity during adsorption and from 95% to 5% relative humidity during desorption. The equilibrium criteria were set at 0.01% weight change in 1 minute with a maximum equilibrium time of 180 minutes. Approximately 1-15 mg of sample was used.

[0061] A colorless blade C20H28C12FN5O4, with approximate dimensions of 0.03 mm x 0.12 mm x 0.50 mm, was used for X-ray crystallographic analysis. X-ray intensity data were measured at 100(2) K on a Bruker Kappa APEX II system equipped with a graphite monochromator and a fine-focus Cu Kα tube (k = 1.54178A), operating at 1.2 kW (40 kV, 30 mA). The detector was placed at a distance of 5.0 cm. from the crystal.

[0062] 7824 frames were collected with a scan width of 0.5° in co and ф and an exposure time of 90 sec / frame. The total data acquisition time was 260 hours. The frames were integrated with the Bruker SAINT software package using the narrow frame integration algorithm. Data integration using a Triclinic cell yielded a total of 12349 reflections up to a maximum th angle of 69.57° (resolution 0.83A), of which 4046 were independent (redundancy 3.06), completeness = 93.6%, Rmt = 5.13%, R S and g = 5.18%) and 3351 (82.8 %) were greater than >2sigma(I) 0 (F 2 ). End cells constant a = 5.9979(4)A, b = 13.4375(9)A, c = 14.4250(9)A, a= 97.617(4)°, 3=93.285(4)°, y= 94.585(5)°, volume = 1145.95(13)A 3, are based on an XYZ-centroid refinement of 4790 reflections above 20 o(I) with 6.196° < 20< 138.239° . Data analysis showed negligible decay during data acquisition. The data were corrected for absorption effects using the multiscan technique (SADABS). The ratio of minimum to maximum apparent transmission was 0.350. The calculated minimum and maximum transmission coefficients (based on crystal size) are 0.3206 and 0.9168.

[0063] The structure was solved and refined with the Bruker SHELXTL software package (Version 6.1), using the space group P-1, with Z = 2 for the formula unit, C20H28Cl2FN5O4. Final anisotropic least squares refinement with full matrices on F 2 with 320 variables converted to R1 = 6.43%, for the observed data and wR2 = 19.18% for all data. The goodness of fit was 1.067. The largest peak in the electron density synthesis was the largest peak 1.084 e7A 3 and the largest hole was -0.527 e / A 3with an RMS deviation of 0.101 e7A 3 Based on the final model, the density was calculated to be 1,427 g / cm 3 and F(000), 516 e“.

[0064] Two partial water occupancies were found and refined in this structure. The water occupancies were independently refined to 53% and 41% for a total water content of 0.94 water equivalents per omecamtiv mecarbil molecule. This is consistent with other measurements of water content in this form of the compound. The hydrogen atoms in one of the solvated waters, the one with 41% occupancy, were found in the electron density difference map and refined with bond lengths fixed at 1.01 Å. Hydrogen atoms at N3, C4, and N4 were found and allowed to refine isotropically. All other hydrogen atoms were placed in idealized positions and refined in a riding manner.

[0065] X-ray powder diffraction (XRPD) data were obtained using a PANalytical X'Pert PRO diffractometer (PANalytical, Almelo, The Netherlands) equipped with a real-time multiple-strip detector (RTMS). Cu Kα (1.54 A) radiation was used, and the voltage and current were set to 45 kV and 40 mA, respectively. Data were collected at room temperature from 5 to 45 degrees 2-theta, with a step size of 0.0334 degrees. Samples were prepared on a low-background sample holder and mounted on a sample stage that was rotated at 2 seconds of rotation.

[0066] Alternatively, XRPD data were obtained using a PANalytical X'Pert PRO diffractometer (PANalytical, Almelo, The Netherlands) equipped with an RTMS detector. Cu Kα(1.54 A) radiation was used, and the voltage and current were set to 45 kV and 40 mA, respectively. Data were collected at room temperature from 5 to 40 degrees 2-theta, with a step size of 0.0334 degrees. Samples were prepared on a low-background sample holder and mounted on a sample stage that was rotated at 2 seconds of rotation.

[0067] Alternatively, XRPD data were obtained using a PANalytics X'Pert PRO diffractometer (PANalytics, Almelo, The Netherlands) equipped with an RTMS detector. Cu Kα(1.54 A) radiation was used, and the voltage and current were set to 45 kV and 40 mA, respectively. Data were collected at room temperature from 5 to 40 degrees 2-theta, with a step size of 0.0167 degrees. Samples were prepared on a low-background sample holder and mounted on a sample stage that was rotated at 2 seconds of rotation.

[0068] Alternatively, XRPD data were obtained using a PANalytics X'Pert PRO diffractometer (PANalytics, Almelo, The Netherlands) equipped with an RTMS detector. Cu Kα radiation (1.54 A) was used, and the voltage and current were set to 45 kV and 40 mA, respectively. Data were collected at room temperature from 3 to 40 degrees 2-theta, with step sizes of 0.008 degrees. Samples were prepared on a low background sample holder and placed on a sample stage with 2 second revolutions.

[0069] Alternatively, XRPD data were obtained using a Bruker D8 X-ray diffraction system (Bruker, Billerica, MA) equipped with a motorized xyz sample stage and a GADDS area detector. Cu Kα radiation (1.54 A) was used, and the voltage and current were set to 45 kV and 40 mA, respectively. Solid samples on a flat glass plate were mapped and for each sample an area of ​​1 mm 2 was scanned in an oscillating mode for 3 minutes from 5 to 48 degrees 2-theta.

[0070] Differential scanning calorimetry (DSC) data were collected using the standard™ DSC mode (DSC Q200, TA Instruments, New Castle, DE). A heating rate of 10°C / min was used for temperatures from 40°C to 300°C. The analysis was performed under nitrogen and the samples were placed in standard, hermetically sealed aluminum containers. Indium was used as a calibration standard.

[0071] Alternatively, DSC data were collected using a temperature modulated DSC mode (DSC Q200, TA Instruments, New Castle, DE). After equilibrating the sample at 20°C for five minutes, a heating rate of 3°C / min was used with a modulation of + / -0.75°C / min over a temperature range of 20°C to 200°C. The analysis was performed under nitrogen and the samples were placed in standard aluminum containers without injection. Indium was used as a calibration standard. Production of Omekamtiv Mecarbyl dihydrochloride hydrate A synthetic route to Omekamtiv McCarbil

[0072] i) iPr2NEt. THF ii) solvent substitution for IPA iii) HCl. H2O PhO SM-2 Phenyl Carbamate-HCl H3CO' omekativ mecarbit-ZHCl-HjO V CH3 • 2HC! • H2O Synthesis of API SM Piperazine Nitro-HCl

[0073] AcOH FN-Toulen National Bank of Serbia Bz2O Piperazine Carboxylate 0 88% in general

[0074] In a 60 L reactor (containing no stainless steel, Hastelloy® H or other metal de love) equipped with a reflux condenser and a scrubber filled with 5N NaOH solution, a mechanically stirred mixture of FN-toluene (2.0 kg, 12.89 mol, 1.0 equiv.), N-bromosuccinimide (3.9 kg, 21.92 mol, 1.70 equiv.), benzoyl peroxide (125.0 g, 0.03 equiv., 0.39 mol, containing 25 wt % water), and acetic acid (7.0 L, 3.5 vol.) was heated at 85 °C under nitrogen for 7 h. A solution of H3PO3 (106.0 g, 1.29 mol, 0.1 equiv.) and acetic acid (200 mL, 0.1 vol.), prepared in a separate vessel, was added. The reaction mixture was stirred for 0.5 h, and analysis of an aliquot confirmed complete decomposition of benzoyl peroxide (not detected, HPLC254nm). The reaction mixture was cooled to 22 °C. DI water (8.0 L, 4 vol.) and toluene (16.0 L, 8 vol.) were charged, the biphasic mixture was stirred (20 min), and the layers were separated. Aqueous 1.6 N NaOH (14.0 L, 7.0 vol) was added to the organic layer at a rate that kept the batch temperature below 25 °C and the pH of the resulting aqueous phase was measured (> 11). The biphasic mixture was filtered through a 5 µm Teflon® cartridge and the layers were separated. The filter line was washed with an additional 2 L of toluene.

[0075] The assay yields were 2.5% FN-toluene, 62.3% FN-bromide, and 30.0% Di-Bromide. The toluene solution contained no benzoyl peroxide, succinimide, or α-bromoacetic acid and water, as determined by KF titration, was 1030 ppm (This solution can be kept under nitrogen at room temperature for >12 h without altering the assay yield).

[0076] To this solution at room temperature was added diisopropylethylamine (880.0 g, 6.63 mol, 0.53 eq.), followed by methanol (460 mL, 11.28 mol, 0.88 eq.) and heated to 40 °C. A solution of diethylphosphite (820.0 g, 5.63 mol, 0.46 eq.) in methanol (460 mL, 11.28 mol, 0.88 eq.) was prepared and added to the reaction mixture at 40 °C via an addition funnel over 1 hour at such a rate that the batch temperature was within 40 ± 5 °C. The contents were stirred for 3 hours at 40 °C from the start of the addition, cooled to room temperature and kept under a nitrogen atmosphere for 12 hours. The yield of the reaction mixture was 2.5% FN-toluene 92.0% FN-bromide and 0.2% Di-Bromide. This solution was used as such for the alkylation step.

[0077] Characteristics for the components of the final product mixture (collected for pure compounds).

[0078] 2-Fluoro-3-Nitrotoluene (FN-Toluene): 11 NMR (400 MHz, CHLOROFORM-7) b ppm 2.37 (s, 1 H), 7.13-7.20 (m, 1 H), 7.45-7.51 (m, 1 H), 7.79-7.85 (m, 1 H). 13C NMR (100 MHz, CHLOROFORM-tZ) b ppm 14.3 (d, J= 5 Hz), 123.3 (d, J= 3 Hz), 123.6 (d, J= 5 Hz), 128.2 (d, J= 16 Hz), 136.7 (d, 7), (15 Hz), (35 Hz). 153.7 (d, J = 261 Hz); 1-(bromomethyl)-2-fluoro-3-nitrobenzene (FN-Bromide): 'H NMR (400 MHz, CHLOROFORM-d) b ppm 4.56 (s, 1 H), 7.28-7.34 (m, 1 H), 7.69-7.76 (m, 1 H-8), 7.98 (Hm). 13 C NMR (100 MHz, CHLOROFORM-J) b ppm 23.6 (d, J= 5 Hz), 124.5 (d, J= 5 Hz), 126.1 (d, J= 3 Hz), 128.5 (d, J = 14 Hz), 136.5 (d, J = 4 Hz), 137.7. (d, J = 265 Hz). DSC: one melting point at 53.59 °C. The mass tray [CvHsBrFNCh + H + : calc. = 233.9566, measured = 233.9561; l-(dibromomethyl)-2-fluoro-3-nitrobenzene (Dibromide): 'll NMR (400 MHz, CHLOROFORM-7) b ppm 6.97 (s, 1 H), 7.39-7.45 (m, 1 H), 8.03-8.10 (m, 1 H), 8.03-8.16 (m, 1 H). 13C NMR (100 MHz, CHLOROFORM-7) b ppm 29.2 (d, J = 7 Hz), 124.9 (d, J = 5 Hz), 127.1 (d, 7= 2 Hz), 132.1 (d, J= 11 Hz), 135.7 (d, J= 2 Hz), 137.2 (broad), 149.8 (d, 7= 2 Hz). 266 Hz). DSC: melt once at 49.03 °C. Exact mass [CvHflBriFNCh + H] + : calc. = 311.8671, measured = 311.8666. Piperazine Nitro-HCl:

[0079] Diisopropylethylamine (1.90 kg, 14.69 mol, 1.14 eq.) was charged to a mechanically stirred toluene solution (9 volumes) of FN-bromide (prepared from the previous step) in a 60 L reactor at 22 °C under a nitrogen atmosphere. To this mixture was added a solution of piperazine carboxylate methyl ester (Piperazine carboxylate) (2.03 kg, 14.05 mol, 1.09 equivalents) in toluene (1.0 L, 0.5 vol) at a rate that allowed the batch temperature to remain below 30.0 °C (exothermic). During the addition, the jacket temperature was adjusted to 5 °C to maintain the batch temperature below 30 °C. The mixture was stirred at 22 °C for 3 h and aliquot analysis confirmed the completion of the alkylation reaction (<1.0 LCAP FN-bromide, HPLC254nm). The reaction mixture was treated with aqueous NH4Cl (20 wt%, 10.0 L, 5 vol; prepared from 2.0 kg NH4Cl and 10.0 L DI water), the biphasic mixture was stirred (30 min), and the layers were separated. The organic layer was washed successively with aqueous NH4Cl. NaHCOs (9 wt%, 10.0 L, 5 volumes; prepared from 0.90 kg NaHCOa and 10.0 L DI water). The organic layer was filtered through a 5 pm Teflon® cartridge and transferred to a drum, washed with filtered water with another 1.0 L toluene and a combined toluene solution (10.0 volumes), weighed and assayed (HPLC) to determine the amount of Piperazine Nitro free base. The assay yield for Piperazine-nitro-base was 89.0%, FN-toluene 2.5% and FN-bromide 0.2% with FN-bromide. The total loss of product in the aqueous wash was <1.0%. This solution was stable under nitrogen for more than 12 h.

[0080] To a mechanically stirred toluene solution of Piperazine Nitro free base, prepared as described above, at 22 °C in a 60 L reactor under nitrogen, IPA (19.4 L, 9.7 vol) and DI water (1.0 L, 0.5 vol) were charged. The mixture was heated to 55 °C and 20% of 1.4 equiv. conc. HCl (Titrated prior to use and charged based on titer value; 276.0 ml, 3.21 mol was charged). The contents were stirred for 15 minutes and Piperazine Nitro-HCl seed (130.0 g, 0.39 mol, 0.03 equiv.) was slurried in IPA (400 mL, 0.2 vol). The mixture was stirred for 30 min and the remaining concentration. HCl (80% charge, 1.10 L, 12.82 mol) was added over a period of 4 h. The mixture was stirred at 55 °C for 1 h, cooled to 20 °C linearly over 1.5 h, and stirred at this temperature for 12 h. The supernatant Piperazine Nitro-HCl concentration was measured (2.8 mg / g). The mixture was filtered through an aurora filter fitted with a 5 μm Teflon® cloth. The thyme was transferred to a clean drum and analyzed. The filter cake was washed twice with IPA (11.2 L, 5.6 cc) and dried to constant weight (defined as < 1.0% weight loss for 2 consecutive TGA measurements over a 2-hour period) on a vacuum filter with nitrogen purge (14 h). The combined losses of Piperazine Nitro-HCl in the mother liquor and in the wash were 2.5%. Piperazine Nitro-HCl was isolated in 3.59 kg in 87.6% corrected yield with > 99.5 wt% and 99.0% LCAP purity.

[0081] Methyl 4-(2-fluoro-3-nitrobenzyl)piperazine-1-carboxylate hydrochloride (Piperazine Nitro-HCl): 1 H NMR (300 MHz, DMSO-J) b ppm 3.25 (n, s, 3 H), 3.52-3.66 (m, 8 H), 4.47 (s, 2 H), 7.44-7.63 (t, 1 H, J= 8 Hz), 7.98-8.15 (m, 1 H), 8.17-8.34 (m, 1 H). 13 C NMR (75 MHz, DMSO-ćZ) b ppm 50.3, 51.4, 52.8, 119.6 (d, J= 14 Hz), 125.1 (d, J= 5 Hz), 127.9, 137.4 (d, J = 8 Hz), 139.8 (d, J = 3 Hz), 152.2, 154.7, 155.7. DSC: onset of melting at 248.4 °C. Exact mass [C13H16FN3O4 + H] + : calculated = 298.1203, measured = 298.1198. Alternative processes for the synthesis of Piperazine Nitro:

[0082] jijissulation mc-tansuHbam ptperadn-l-mdilktnboksiJat nieryl ^(i'fluoKv^nitrobenzHjpiperaziivl'carboxylate hydrokknide

[0083] A mixture of NaBH4 (1.7 g, 44 mmol) in THF (68 mL) was treated with 2-fluoro-3-nitrobenzoic acid (3.4 g, 18.4 mmol) and cooled to 0-5 °C. Then the iodine solution was added drop by drop (4.7 g, 18.4 mmol) in THF (12 mL) at a rate to control gas evolution. The progress of the reaction was assessed by HPLC. After 2 h, HPLC assay showed that 4% AUC of 2-fluoro-3-nitrobenzoic acid remained. The mixture was quenched with 1 M HCl (30 mL) and extracted with MTBE (5 mL). The organic layers were then washed with 20% aqueous KOH and 10% sodium thiosulfate. The organic layers were dried with Na2SO4, filtered through Celite, and concentrated to give (2-fluoro-3-nitrophenyl)methanol (2.8 g, 88%, 89% AUC by HPLC).

[0084] A solution of (2-fluoro-3-nitrophenyl)methanol (2.8 g, 16 mmol) in 2-MeTHF (26 mL) was treated with triethylamine (4.5 mL, 32 mmol) and cooled to 0-5 °C. The solution was then treated with methanesulfonyl chloride (1.6 mL, 21 mmol). The progress of the reaction was assessed by HPLC. After 30 minutes at 0-5 °C, the reaction was considered complete. The mixture was quenched with water (14 mL) and the phases were separated. The organic layers were washed with brine, dried over Na2SO4, filtered through Celite and concentrated to give 2-fluoro-3-nitrobenzyl methanesulfonate (3.3 g, 83.1%, 81% AUC by HPLC) as a yellow oil.

[0085] A solution of 2-fluoro-3-nitrobenzyl methanesulfonate (3.3 g, 13 mmol, AMRI lot # 46DAT067B) in toluene (33 mL) was treated with diisopropylethylamine (2.7 mL, 15 mmol) in one portion. A solution of methylpiperazine-1-carboxylate (2.1 g, 15 mmol) in toluene (1.1 mL) was added slowly via syringe to maintain between 23-29 °C. The reaction was stirred for 16 hours after addition. HPLC assay after this time indicated that the reaction was complete. 20% aqueous NH4Cl (11 mL) was added at 20-25 °C. The biphasic mixture was stirred for 15 minutes and the phases were separated. This procedure was repeated using 9% aqueous sodium bicarbonate (11 mL). The toluene layer was then filtered through Celite at 20-25 °C. 2-Propanol (50 mL) and water (1.1 mL) were added to the toluene solution and the mixture was heated to 55-60 °C. The mixture was then treated with 37 wt % HCl (1.6 mL, 18.7 mmol) for 20 min. The addition of a precipitate was observed after the addition.Once the addition was complete, the mixture was allowed to gradually cool to 20-25 °C and stirred for hours before filtering and washing with IPA (2 bed volumes).

[0086] The cake was then dried under vacuum to give 4-(2-fluoro-3-nitrobenzyl)piperazine-1-carboxylate hydrochloride (2.41 g, 54%, 90% AUC by HPLC, 88 wt% by HPLC). Piperazine Nitro Freebase:

[0087] In a 60 L reactor equipped with a reflux condenser, a mixture of piperazine nitro-HCl (2.0 kg, 5.99 mol, 1.0 eq.) and isopropyl acetate (6.0 L, 3.0 vol.) was mechanically stirred at room temperature under nitrogen. A solution of sodium bicarbonate (629 g, 7.49 mol, 1.25 eq.) and water (7.5 L, 3.75 vol.), prepared in a separate vessel, was added. The biphasic mixture was stirred (15 min.), and the layers were separated. The upper organic layer (containing the product) was transferred to a separate vessel, while the reactor was rinsed with water and isopropanol. The organic layer was then passed through a 5 µm Teflon® cartridge back into a clean 60 L reactor. The filter line was rinsed with 4.0 L (2.0 vol.) of isopropanol into the 60 L reactor. An additional 12.0 L (6.0 volumes) of isopropanol was added to the 60 L reactor and heated to 40 °C. Under reduced pressure (67 mbar (50 torr)) the batch was concentrated to approximately 6 L (3.0 volumes). The solution was cooled from 27 °C to 20 °C linearly over 10 minutes. Water (4.0 L, 2.0 vol) was added at 20 °C over 30 min, followed by Piperazine Nitro Freebase seed (18 g, 0.06 mol, 0.01 equivalent). The mixture was aged for 5 min, and the remaining water (24.0 L, 12.0 vol) was added over 90 min. After standing overnight at 20 °C, the supernatant Piperazine Nitro Freebase concentration (<10 mg / mL) was measured. The mixture was filtered through an aurora filter fitted with a 12 pm Teflon® cloth. The filter cake was washed with a mixture of water (3.3 L, 1.65 vol) and isopropanol (700 ml, 0.35 vol) and dried to constant weight (defined as < 1.0% weight loss for 2 consecutive TGA measurements over a 2 h period) on a vacuum filter with nitrogen purge (48 h). The combined losses of Piperazine Nitro Freebase in the mother liquor acids and washing were about 7.5%. Piperazine Nitro Freebase was isolated in 1.67 kg in 92.5% corrected yield with 100.0 wt% and 99.4% LCAP purity. Synthesis of API SM Phenyl Carbamate-HCl

[0088] ^u СНз CICO2Ph H2N^ N AON, NMP Amino pyridine

[0089] A 60 L, glass-capped reactor was set at 20 °C under a nitrogen atmosphere and irradiated through a scrubber (containing 5N NaOH) was charged with 2.5 kg of Amino Pyridine (1.0 equiv., 23.1 mol) and then with 25 L (19.6 kg, 10 vol.) of acetonitrile. After stirring was initiated and the (endothermic) dissolution of the Amino-pyridine was completed, the vessel was charged with 12.5 L of N-methyl-2-pyrrolidinone (12.8 kg, 5 vol.). An addition funnel was charged with 1.8 L (0.6 equiv., 13.9 mol) of phenyl chloroformate which was then added to the Amino Pyridine solution over 68 min while maintaining the internal temperature < 30 °C. The reaction was stirred for 30 min at an internal temperature of 20 ± 5 °C. The vessel was then charged with 61 ± 1 g of seeds as a suspension in 200 ml of acetonitrile and aged for > 30 min. The addition funnel was charged with 1.25 L (0.45 equivalents, 9.7 moles) of phenyl chloroformate, which was then added over 53 minutes to the reaction slurry, again maintaining a temperature of <30 °C. The contents of the reactor were aged > 30 hours at 20 ± 5 °C.After screening the supernatant (< 15 mg / g for both product and starting material), the solids were filtered using an Aurora filter equipped with a 12µm Teflon pad. The mother liquor was transferred to a second 60 L, glass reactor. The reactor and cake were washed with 1 x 10 L of 5:10 NMP / ACN and 1 x 10 L ACN. The washings were also transferred to a second reactor. The cake was dried under vacuum, under nitrogen for > 24 hours, to afford 5.65 kg (90.2% yield) of the product, phenyl carbamate-HCl as an off-white solid at 98.8% by weight with 99.2% LCAP purity.

[0090] Phenyl (6-methylpyridin-3-yl)carbamate hydrochloride (Phenyl Carbamate-HCl) J H NMR (400 MHz, DMSO-b / b) b ppm 11.24 (s, 1 H), 8.81 (s, 1 H), 8.41 (d, 1 H, J = 8.8 Hz), 7.85 (d, 1 H, J = 8.8 Hz), 7.48 - 7.44 (m, 2 H), 7.32 - 7.26 (m, 3 H), 2.69 (s, 3H); 13C NMR (100 MHz, DMSO-t / б) б ppm 151.66, 150.01, 147.51, 136.14, 133.79, 129.99, 129.49, 127.75, 125.87, 121.70, 18.55: HR-MS: Calcd for C13H12N2O2: 228.0899, M + H + = 229.0972; Observed mass: 229.0961 Alternative synthesis of phenyl carbamate HC1

[0091] 5-Amino-2-methylpyridine (53.2 kg, 1.0 equiv.) and acetonitrile (334 kg, 8.0 ml / g) were charged to a nitrogen-filled glass reactor. The reactor contents were stirred while warming to 25-30 °C. The mixture was then recirculated through a filter packed with activated carbon (11 kg, 20 wt.%) at 3 hour intervals while maintaining 25-30 °C. After each 3 hour interval, a sample of the mixture was analyzed for color by comparison with a color standard and UV absorbance at 440 nm. Once a satisfactory result was achieved, the filter was blown into the reactor and the filter was washed with acetonitrile (85 kg, 2.0 ml / g). The acetonitrile wash was transferred to the reaction mixture. 1-Methyl-2-pyrrolidinone (274 kg, 5.0 mL / g) was charged to the reaction mixture in the reactor with glass layers. Phenyl chloroformate (46.6 kg, 0.6 eq.) was slowly added to the mixture while maintaining 15-30 °C (typically 60-70 min). The reaction mixture was stirred for approximately 60 minutes while maintaining 20-25 °C. Chemical crystals of phenyl(6-methylpyridin-3-yl)carbamate hydrochloride (0.58 kg, 0.010 eq.) were added to the stirring mixture. The suspension was then stirred for approximately 4 hours at 20 ± 5 °C. Phenyl chloroformate (33.4 kg, 0.45 eq.) was slowly added to the slurry while maintaining 15-30 °C. The mixture was then allowed to age with stirring for 8 ± 1 h, after which it was concentrated. 5-amino-2-methylpyridine (target value <15 mg / mL) and phenyl (6-methylpyridin-3-yl)carbamate hydrochloride (target value <15 mg / mL) were checked by HPLC. The batch was then filtered under vacuum and washed with a mixture of acetonitrile (112 kg, 2.68 mL / g) and 1-methyl-2-pyrrolidinone (72 kg, 1.32 mL / g), followed by washing with acetonitrile (167 kg, 4.0 mL / g).The solid residue was separated and then transferred to a bed dryer maintained between 20-40 °C and 90-44 mbar (1.3-0.65 psia) until an LOD of <1 wt% was achieved, after which phenyl(6-methylpyridin-3-yl)carbamate hydrochloride 106.3 kg (81.6% yield) was isolated from the dryer. Methyl 4-(3-amino-2-fluorobenzyl)piperazine-1-carboxylate (Piperazine Aniline)

[0092] Neutralization + CO г (And equiv.) + H г O (1 equiv.) + NaHCO3 <0.25 equiv.) where 60 psig corresponds to 4.13 barg.

[0093] In a 100 L glass-capped reactor, methyl 4-(2-fluoro-3-nitrobenzyl) piperazine-1-carboxylate hydrochloride (2.00 kg, 1.00 eq.) and isopropyl acetate (6.00 L, 3.00 eq. based on starting material) were added. The resulting suspension was stirred under nitrogen. To the mixture was added dropwise over 45 ± 30 minutes: 7.7% w / w aqueous sodium bicarbonate solution (629 g, 1.25 equivalents of sodium bicarbonate dissolved in 7.50 L of water), maintaining an internal temperature of 20 ± 5 °C by jacket control (NOTE: the addition is endothermic and may evolve up to 1 equivalent of carbon dioxide gas). The mixture was stirred for > 15 minutes, resulting in a clear two-phase mixture. Stirring was stopped and the layers were allowed to settle.

[0094] The lower (aqueous) layer was dried and analyzed using pH paper to ensure that the layer pH was > 6. Quantitative HPLC analysis of the upper (organic) layer showed 97-100% assayed yield of methyl 4-(2-fluoro-3-nitrobenzyl)piperazine-1-carboxylate free base (1.73 - 1.78 kg). The upper (organic) layer was transferred through an in-line filter to a 20-liter Hastelloy® hydrogenator, and the 100-L reactor and lines were rinsed with an additional aliquot of isopropyl acetate (2.00 L, 1.00 cc). The hydrogenator was purged with nitrogen and vented to atmospheric pressure. A suspension of 5.0 wt % palladium on carbon (20.0 g, Strem / BASF Escat™ 1421, approximately 50% water) in isopropyl acetate (400 mL) was added to the reaction mixture, followed by 400 mL of washing. The resulting reaction mixture was diluted with an additional aliquot of isopropyl acetate (1.2 L; total isopropyl acetate volume 10.0 L, 5.00 cc). The hydrogenator was purged with nitrogen three times (pressured to 4.13 ± 0.69 barg (60 ± 10 psig), then vented to atmospheric pressure), and then pressurized to 4.13 ± 0.34 barg (60 ± 5 psig) with hydrogen. The reaction mixture was stirred at <100 rpm at 30 ± 5 °C while maintaining 4.13 ± 0.34 barg (60 ± 5 psig) of hydrogen, for 2 hours until the reaction was considered complete. This temperature and pressure corresponded to a measured kLa value of about 0.40 in a 20-L hydrogenator. The end of the reaction was determined by a dramatic decrease in hydrogen consumption accompanied by a decrease in the heat of the reaction. To control for potential dimer impurities, the reaction was continued for at least 30 minutes after this change in reaction profile and HPLC analysis was performed to confirm that >99.5% conversion of hydroxylamine to aniline had been achieved.

[0095] At the end of the reaction, the hydrogenator was purged twice with nitrogen (pressurized to 4.13 ± 0.69 barg (60 ± 10 psig) and then vented to atmospheric pressure). The crude reaction mixture was filtered through a 5 µm filter and then a 0.45 µm filter in series into a 40-L glass-lined reactor. The hydrogenator and lines were rinsed with an additional aliquot of isopropyl acetate (2.00 L). Quantitative HPLC analysis of the crude reaction mixture indicated 95-100% assay yield (1.52 to 1.60 kg aniline product). The reaction mixture was distilled under reduced pressure (typically 250 - 300 mbar) at a batch temperature of 50 ± 5 °C until the total reaction volume was approximately 8.00 L (4.00 cfm). The batch was subjected to constant volume distillation at 50 ± 5 °C, 250 - 300 mbar, adding heptane to control the total batch volume. After approximately 8.00 L (4.00 cc) of heptane, GC analysis showed that the solvent composition was approximately 50% isopropyl acetate, 50% heptane. The vacuum was broken, and the internal temperature of the batch was maintained at 50 ± 5 °C. A seed suspension (20.0 grams of the product methyl 4-(3-amino-2-fluorobenzyl) piperazine-1-carboxylate, in a solvent mixture of 80 ml heptane and 20 ml isopropyl acetate) was added to the reaction mixture. The resulting suspension was allowed to stir at 50 ± 5 °C for 2 ± 1 h, then cooled to 20 ± 5 °C over 2.5 ± 1.0 h. Additional heptane (24.0 L, 12.0 g) was added dropwise over 2 hours, and the batch was allowed to stir at 20 ± 5 °C for > 1 hour (usually overnight). Quantitative HPLC analysis of this filtered supernatant revealed < 5 mg / mL of product in solution, and the product crystals were double rods of 50 to 400 gm.The reaction suspension was filtered at 20 °C on a filter cloth, and the cake was washed with heptane (6.00 L, 2.00 t). The cake was dried on the filter under nitrogen for 4 hours until the dry sample was confirmed by LOD analysis (indicated <1.0 % mass loss). The product methyl 4-(3-amino-2-fluorobenzyl)piperazine-1-carboxylate (1.56 kg) was isolated as a pale yellow powder in 86% yield at 99.8 % mass by HPLC with 100.0 LCAP210. [Analysis of the combined filtrates and washings revealed 108 grams (7.0%) of product lost to the mother liquor. The remaining mass mass consisted of product retention in the reactor (fouling).] NMR (DMSO- < / 6, 400 MHz) б: 6.81 (dd, J = 7.53, 7.82 Hz, 1H), 6.67 (m, 1H), 6.49 (m, 1H), 5.04 (s, 2H), 3.58 (s, 3H), 3.45 (m, 2H), 3.34 (m, 4H), 2.33 (m, 4H). . 19 F NMR (d6-DMSO, 376 MHz) b: - 140.2. 13 C NMR (d 6 -DMSO, 125 MHz) b: 155.0, 150.5, 148.2, 136.2 (m), 123.7 (m), 117.6, 115.1, 73.7, 54.9 (m), 52.1 (m), 43.4. mp = 89.2 °C. An alternative route to piperazine aniline

[0096] Methyl 4-(2-fluoro-3-nitrobenzyl)piperazine-1-carboxylate hydrochloride (46.00 kg, 1.00 eq.) and isopropyl acetate (200 kg, 5.0 ml / g) were added to a glass-capped reactor. The resulting suspension was stirred under nitrogen. A 7.4% w / w aqueous sodium bicarbonate solution (1.25 eq.) was added to the mixture while maintaining the internal temperature at 25 ± 5 °C. The mixture was stirred for > 30 minutes, resulting in a clear two-phase mixture. Stirring was stopped and the lower (aqueous) layer was discarded. Analysis of the aqueous layer showed a pH of > 6. Water (92 kg, 2.0 ml / g) was charged to the organic layer and stirred for 15 minutes. Stirring was then stopped and the lower (water) layer was discarded. Water (92 kg, 2.0 ml / g) was charged into the organic layer and stirred for 15 minutes. Then the stirring was stopped and the bottom layer (water) was drained. The batch was distilled under reduced pressure while maintaining the batch temperature between 40- 50 °C. The batch volume was kept constant during the distillation by continuous addition of isopropyl acetate. Once the batch water content was <1500 ppm, the solution was transferred through an in-line filter to a Hastelloy reactor containing 5.0 wt.% palladium on carbon (BASF Escat 1421, 0.69 kg, 1.5 wt.%). The glass-capped reactor was rinsed with isopropyl acetate (100 kg, 2.5 ml / g) and added to the Hastelloy reactor using a line filter.

[0097] The batch was adjusted to approximately 25-35 °C (preferably 30 °C) and hydrogen gas was added to maintain about 4 barg with vigorous stirring. Hydrogenation was continued for 1 hour after the hydrogen feed was stopped and conversion >99.0% was achieved by HPLC. The palladium on carbon catalyst was collected by filtration and the supernatant was collected in the reactor. Isopropyl acetate (40 kg, 1.0 ml / g) was charged to the Hastelloy reactor and passed through the filter and collected in the in-line glass reactors.

[0098] The batch was concentrated under reduced pressure while maintaining the batch temperature between 35-55 °C until the final volume was approximately 4.0 ml / g. Heptane (219 kg, 7.0 mL / g) was added to the glass-capped reactor while maintaining the batch temperature between 50-60 °C, until 20-25% isopropyl acetate in heptane was achieved as measured by GC. The solution was cooled to a temperature between 40-50 °C and seeded with methyl 4-(3-amino-2-fluorobenzyl) piperazine-1-carboxylate (0.46 kg, 1.0 wt%) as a suspension in heptane (6.4 kg, 0.20 ml / g). The slurry was aged for approximately 2 hours, after which the batch was distilled under reduced pressure while maintaining the batch temperature between 35-45 °C. The batch volume was kept constant during the distillation by continuous addition of heptane (219 kg, 7.0 rnL / g). The batch was then cooled to between 15-25 °C over approximately 3 h. The supernatant concentration was measured to be <5 mg / mL methyl 4-(3-amino-2-fluorobenzyl)piperazine-1-carboxylate by HPLC.

[0099] The batch was filtered and the resulting solids were washed successively with heptane (63 kg, 2.0 ml / g), then heptane (94 kg, 3.0 ml / g). The solids were dried on the filter by a stream of dry nitrogen under vacuum until an LOD of < 1 wt% was achieved, after which 33.88 kg (90.7% yield) was isolated from the filter drier. Procedure Omekamtiv Mecarbyl Dihydrochloride hydrate

[0100] (1.0 cc) Piperazine Analine rnwi 4-(3-aw. <no-2-fluorobe-nzii)pipera2in.-1 karboksilai (1.2 equiv.) Phenyl Carbamate-HCl fcmi ((vmctilpirklin- 3-yl)carban.ia$ hydrochloride DiPEA 0 30 dcviv.) THE (4V), 65 C, 8-24 h DIPEA-HC1 (L2 equiv.) + DIPEA(O.lOtfkiv.) Phenol ((.0 equiv.) 2539850(02 ckvK.) 1) 2-PrOH(11 V) 2) De-scratch up to 4V 3) Water (2.30 V) 4) 6N NSC2.4 equiv.) 5) 2-PrOH (16.5V) 6) Wet Mill *2HCi-H ? About

[0101] A 15 L glass reactor was charged with methyl 4-(3-amino-2-fluorobenzyl)piperazine-1-carboxylate (1.202 g, 4.50 mol), phenyl(6-methylpyridin-3-yl)carbamate hydrochloride (1.444 g, 5.40 mol), and tetrahydrofuran (4.81 L). The resulting suspension was stirred under nitrogen and N,N-diisopropylethylamine (1.019 L, 5.85 mol) was then charged into the suspension resulting in a brown solution. The solution temperature was increased to 65 °C and stirred for 22 h, until <1% AUC piperazine aniline remained by HPLC analysis.

[0102] The batch was cooled to 50 °C and distilled under reduced pressure while maintaining the internal temperature of the vessel below 50 °C by adjusting the vacuum pressure. 2-propanol was added with residual vacuum at a rate to maintain a constant volume in the 15 L reactor. A total of 10.5 kg of 2-propanol was required to achieve <5% THF by GC. Water (2.77 kg) was then charged to the reactor, after which 6N HCl (1.98 kg) was added at a rate to maintain the internal temperature below 60 °C. The reactor was brought to atmospheric pressure under nitrogen. The solution was then heated to 60 °C and transferred to a 60 L glass reactor through an inline filter. The 15L reactor was then rinsed with 1:1 water / 2-propanol (1.2 L) which was sent through an inline filter to the 60L reactor.

[0103] A 60L reactor was set at 45 °C and a suspension of seeds (114 g, 0.23 mol) in 2-propanol (0.35 L) was added to the reactor resulting in a suspension. The batch was aged at 45 °C for 1 hour, after which 2-propanol (3.97 kg) was added through an in-line filter over 2 hours. The batch was heated to 55 °C for 1 h and held for 0.25 h, then cooled back to 45 °C for 1 h and held overnight at 45 °C. 2-propanol (11.71 kg) was then added through an in-line filter over 3 h. The batch was aged for 1 h, then cooled to 20 °C over 2 h and held at 20 °C for 0.5 h. The batch was then recirculated through a wet mill set up with 1-medium and 2-fine rotor-stators operating at 56 Hz for 2.15 h, while no further reduction in particle size was observed by microscopy.

[0104] The batch was then filtered through a 50.8 cm (20 in) Hastelloy® filter fitted with a 12 pm filter cloth under a vacuum of 667 mbar (500 Torr). A 95:5 2-propanol:water wash solution (1.82 L) was charged through the inline filter into a 60 L reactor and then onto the filter. A second 2-propanol wash (2.85 L) was charged through the inline filter into a 60 L reactor and then onto the filter. The batch was then dried under a wet nitrogen pressure of 345 mbar (5 psi) to <5000 ppm 2-propanol, with 2.5-5% water remaining. The final solid was drained from the filter to give 2.09 kg of methyl 4-(2-fluoro-3-(3-(6-methylpyridin-3-yl)ureido)benzyl)piperazine-1-carboxylate as white crystals in 89% yield of 99.88 wt% by HPLC, 100.0% AUC. Total losses from alcoholic beverages were 0.10 kg (4.7%).

[0105] DSC: Tonset = 61.7 °C, T max= 95.0 °C; TGA = 2.2%, higher temperature = 222 °C; 'H HMR(D20, 500 MHz) б 8.87 (s, 1H), 8.18 (d, J = 8.9 Hz, 1H), 7.83 (t, J = 7.5 Hz, 1H), 7.71 (d, J = 8.8 Hz, 1H), 7.35-7.29 (m, 2H), 4.48 (s, 2H), 4.24 (br s, 2H), 3.73 (s, 3H), 3.31 (br s, 6H), 2.68 (s, 3H); 13 C HMR (D2O, 150 MHz) б 156.8, 154.2, 153.9 (7 = 249 Hz), 147.8, 136.3, 136.1, 130.1, 129.4, 128.0, 127.2, 125.5 (7 = 11.8 Hz), 125.1 (7 = 4.2 Hz), 116.1 (7 = 13.5 Hz), 53.54, 53.52, 53.49, 50.9, 40.5, 18.2. Alternativni postupak spajanja (Anilin fenil carbamata)

[0106]

[0107] A reaction vessel was charged with methyl 4-(3-amino-2-fluorobenzyl)piperazine-1-carboxylate (2.5 g, 1.0 eq.), acetonitrile (25.0 mL, 10.0 mL / g), and 1-methyl-2-pyrrolidinone (12.5 mL, 5.0 mL / g). The batch was cooled to 0 °C, after which phenyl chloroformate (1.20 mL, 1.02 eq.) was added over approximately 5 minutes. After 45 minutes, the resulting suspension was allowed to warm to 20 °C. The solids were collected by filtration and washed twice with acetonitrile (10.0 mL, 4.0 mL / g). The solid residues were dried under a stream of dry nitrogen to give methyl 4-(2-fluoro-3-((phenoxycarbonyl)amino)benzyl)piperazine-1-carboxylate hydrochloride 2.8 g (71% yield) as a white solid.

[0108] 4-(2-fluoro-3-((phenoxycarbonyl)amino)benzyl)piperazine-1-carboxylate hydrochloride: JH NMR (400 MHz, DMSO-b / b) b ppm 3.08 (No. s, 2 H), 3.24 - 3.52 (m, 4 H), 3.62 (s, 3 H), 4.03 (d, 7=11.25 Hz, 2 H), 4.38 (No. s, 2 H), 7.11 - 7.35 (m, 4 H), 7.35 - 7.49 (m, 2 H), 7.49 - 7.66 (m, 1 H), 7.80 (s, 1 H), 10.12 (nr. s, 1 H), 11.79 (nr. s, 1 H); HRMS = 388.1676 found, 388.1667 calculated.

[0109] The reaction vessel was charged with methyl 4-(2-fluoro-3-((phenoxycarbonyl)amino)benzyl)piperazine-1-carboxylate hydrochloride (0.50 g, 1.0 equiv.), 6-methylpyridin-3-amine (0.15 g, 1.2 equiv.), tetrahydrofuran (2.0 mL, 4.0 mL / g) and N,N-diisopropylethylamine. (0.23 mL, 1.1 eq.). The batch was heated at 65 °C for 22 hours, after which quantitative HPLC analysis showed 0.438 g (92% test yield) of mekamtiv mekarbil. Alternative Procedure Omekamtiv Mecarbyl Dihydrochloride Hydrate

[0110] Omekamtiv Mecarbil, free base (3.0 kg, 1.0 equiv.), was charged to a nitrogen-filled jacketed vessel, followed by water (4.6 L, 1.5 ml / g) and 2-propanol (6.1 L, 2.60 ml / g). The slurry was stirred and heated to approximately 40 °C, after which 6N HCl (2.6 L, 2.10 equiv.) was added to the slurry resulting in a colorless, homogeneous solution. The solution was heated to between 60-65 °C and transferred through an in-line filter into a 60L reactor preheated to 60 °C. The batch was cooled to 45 °C, after which Omekamtiv Mecarbil dihydrochloride hydrate (150 g, 5.0 wt%) was charged to the vessel as a suspension in 95:5 (v / v) 2-propanol / water (600 mL, 0.20 mL / g). The resulting suspension was maintained at 45 °C for 0.5 h, followed by cooling to approximately 20 °C, followed by holding for 3-16 h. 2-propanol (33.0 L, 11.0 mL / g) was added over >2 h, followed by an isothermal hold of about 1 h at about 20 °C. (Supernatant pH

[0111] The batch was recirculated through a wet mill for 5-10 batch revolutions until sufficient particle reduction was achieved compared to the uncalibrated visual microscopy reference. The suspension was filtered under vacuum, and the resulting solids were washed with two washes of 95:5 (v / v) 2-propanol / water (3.0 L, 1.0 mL / g) and a final cake wash with 2-propanol (6.0 L, 2.0 mL / g). The cake was dried on the filter by blowing wet nitrogen through the cake until < 5000 ppm 2-propanol and 2.5-5% water were measured by GC and KF analysis. Omecamtiv Mecarbyl dihydrochloride hydrate was isolated as a colorless crystalline solid (3.40 kg, 93% yield). pH-dependent release profiles

[0112] A formulation of omecamtiv mecarbil hemihydrate (free base) and dihydrochloride hydrate (Form A) was prepared with the following components, all components reported as w / w%: Free base (75 mg matrix tablet) Active granulation: 15.37% free base; 30% hypromellose, HPMC K100 MPrem CR; 10% citric acid monohydrate; 11.88% microcrystalline cellulose, Avicel PH 101; 6.75% lactose monohydrate, FastFlo 316; 12.5% ​​purified water; and lemon Acid granulation: 20% citric acid monohydrate; 5% microcrystalline cellulose, Avicel PH 101; and 1% magnesium stearate, non-bovine. Form A (75 mg matrix tablet) Intra-granulation: 18.37% Form A; 30% hypromellose, HPMC K100 MPrem CR; 0.50% magnesium stearate; and Extra-granulation: 16.88% microcrystalline cellulose, Avicel PH 101; 18.37% anhydrous citric acid; and 0.5% magnesium stearate, non-bovine.

[0113] The formulations were tested at pH 2 and pH 6.8 and the amount of drug released over time was measured. The results of this drug release profile are shown in Figure 6.

Claims

Patent claims 1. Dihydrochloride hydrate salt softening mekarbil.

2. The salt of claim 1, wherein the dihydrochloride salt is a monohydrate.

3. The salt according to claim 1 or 2, wherein the salt is crystalline.

4. The salt according to any one of claims 1 to 3, characterized in that the salt is characterized by an X-ray powder diffraction pattern containing maxima at 6.6, 14.9, 20.1, 21.4 and 26.8 ± 0.2° 2θ using Cu Kα radiation.

5. The salt of claim 4, wherein the X-ray powder diffraction pattern further comprises maxima at 8.4, 24.2, 26.0 and 33.3 + 0.2° 2θ using Cu Kα radiation.

6. The salt of claim 4 or 5, wherein the X-ray powder diffraction further comprises peaks at 6.2, 9.7, 13.2, 14.3, 15.4, 16.3, 16.9, 18.9, 19.5, 20.7, 21.8, 22.8, 23.6, 25.1, 27.3, 27.7, 28.4, 29.4, 30.2, 31.2, 31.5, 31.9, 33.9, 34.5, 34.9, 36.1, 36.8, 37.7, 38.5, and 39.7+ 0.2° 20 using Cu Kα radiation.

7. The salt according to any one of claims 1 to 6, having an X-ray powder diffraction pattern substantially as shown in Figure 2.

8. A salt according to any one of claims 1 to 7, which has an endothermic transition at 230 °C to 240 °C, measured for example by differential scanning calorimetry, wherein the transition is at 235 °C.

9. A process for preparing omecamtiv mecarbil dihydrochloride hydrate salt comprising mixing methyl 4-(3-amino-2-fluorobenzyl)piperazine-1-carboxylate and phenyl(6-methylpyridin-3-ylcarbamate in the presence of a trialkylamine base to form omecamtiv mecarbil; further comprising crystallizing omecamtiv mecarbil in the presence of aqueous hydrochloric acid and an alcoholic solvent to form omecamtiv mecarbil dihydrochloride hydrate salt, for example, wherein the alcoholic solvent comprises isopropyl alcohol.

10. The method of claim 9, wherein omecamtiv mecarbil dihydrochloride hydrate salt has an X-ray powder diffraction (XRPD) pattern containing peaks at 6.6, 14.9, 20.1, 21.4 and 26.8 ± 0.2° 20, using Cu Kα radiation, optionally wherein the X-ray powder diffraction pattern further comprises peaks at 8.4, 24.2, 26.0 and 33.3 ± 0.2° 20 using Cu Kα radiation and / or optionally wherein the X-ray powder diffraction pattern further comprises maxima at 6.2, 9.7, 13.2, 14.3, 15.4, 16.3, 16.9, 18.9, 19.5, 20.7, 21.8, 22.8, 23.6, 25.1, 27.3, 27.7, 28.4, 29.4, 30.2, 31.2, 31.5, 31.9, 33.9, 34.5, 34.9, 36.1, 36.8, 37.7, 38.5, and 39.7 + 0.2° 20 using Cu Kα radiation.

11. The method according to claim 9, comprising: (a) hydrogenating methyl 4-(2-fluoro-3-nitrobenzyl)piperazine-1-carboxylate in the presence of a hydrogenation catalyst to give methyl 4-(3-amino-2-fluorobenzylpiperazine-1-carboxylate); (b) mixing methyl 4-(3-amino-2-fluorobenzyl)piperazine-1-carboxylate and phenyl(6-methylpyridin-3-yl)carbamate in the presence of a trialkylamine base to form the omecamtiv mecarbil as the free base; and (c) crystallization of omecamtiv mecarbil free base in the presence of aqueous hydrochloric acid and an alcoholic solvent to form omecamtiv mecarbil dihydrochloride hydrate salt.

12. The process according to claim 11, wherein the hydrogenation catalyst comprises palladium, preferably palladium on carbon.

13. The process according to claim 11 or 12, wherein the trialkylamine base is triethylamine, diisopropylethylamine or a combination thereof, preferably wherein the trialkylamine base comprises diisopropylethylamine.

14. The method according to any one of claims 11 to 13, wherein the alcoholic solvent comprises isopropyl alcohol.

15. The method of any one of claims 11 to 14, wherein the omecamtiv mecarbil dihydrochloride hydrate salt has an X-ray powder diffraction pattern (XRPD) comprising maxima at 6.6, 14.9, 20.1, 21.4 and 26.8 ± 0.2° 20 using Cu Kα radiation, optionally wherein the X-ray powder diffraction pattern further comprises maxima at 8.4, 24.2, 26.0 and 33.3 ± 0.2° 20 using Cu Kα radiation, and / or optionally an X-ray powder diffraction pattern pattern that further contains highs at 6.2, 9.7, 13.2, 14.3, 15.4, 16.3, 16.9, 18.9, 19.5, 20.7, 21.8, 22.8, 23.6, 25.1, 27.3, 27.7, 28.4, 29.4, 30.2, 31.2, 31.5, 31.9, 33.9, 34.5, 34.9, 36.1, 36.8, 37.7, 38.5, and 39.7 ± 0.2° 29 using Cu Kα radiation.