EA046380B1 - POLYMORPHOUS FORMS OF COMPOUND A AND THEIR PHARMACEUTICAL COMPOSITIONS - Google Patents

Description

injuries.

Field of technology

The invention relates to compounds and methods of using them for treating, facilitating or accelerating recovery from certain conditions of the brain, central nervous system (CNS) or cardiovascular system, such as brain injury, neurodegenerative condition or cardiac ischemia.

Cross reference to related applications

This application claims priority to U.S. Provisional Application No. 62/736,979, filed September 26, 2018, the contents of which are hereby incorporated by reference in their entirety.

State of the art

Brain injury is unfortunately a common pathological condition and a leading cause of morbidity and mortality worldwide. The brain in particular is susceptible to damage because neurons have limited ability to repair themselves. At the time a subject is born, the brain already contains essentially all the neurons it will have during its lifetime. Unlike other cells in the body, neurons stop reproducing shortly after birth. If these cells become damaged or die, they are not replaced, often resulting in a disabling and largely irreversible decline in the subject's cognitive and sensorimotor abilities. Conditions that lead to the death and damage of nerve cells range from episodes of ischemia (eg, stroke) and injury to degenerative disorders (eg, Alzheimer's disease).

Damage to the central nervous system (CNS) is a significant cause of death and disability worldwide. For example, according to the US Centers for Disease Control and Prevention (CDC), approximately 1.7 million people suffer traumatic brain injury (TBI) each year, costing the US economy more than $60 billion per year in medical costs and lost productivity. labor (Finkelstein, E; Corso, P; Miller, T, The Incidence and Economic Burden of Injuries in the United States, Oxford University Press: New York, 2006). In addition, stroke is the third leading cause of death in the US with an estimated incidence of 795,000 annually, a leading cause of disability, and costs the US economy more than $34 billion annually (NINDS, 2014; strnoke.nih.gov; and Mozaffarian D, Benjamin EJ, Go AS, et al. Heart disease and stroke statistics-2015 update: a report from the American Heart Association, Circulation. 2015; e29-322). In an emergency setting, it is possible to treat within 24 hours, which may limit the extent of the damage. Immediately after an ischemic or hemorrhagic stroke, the stroke site in the brain typically contains a central mass of tissue that is irreversibly damaged, as well as an area of viable but at-risk tissue called the ischemic penumbra. During this period, insufficient supply of oxygen and glucose to brain cells leads to further secondary damage to the penumbra region. Lack of oxygen and glucose reduces energy production by cell mitochondria. The immediate consequence of this lack of energy is the disruption of ion pumps, which, due to increased levels of extracellular potassium ions (K ⁺ ), causes waves of repeated propagating depolarization in brain tissue. At the same time, the influx of sodium ions (Na ⁺ ) into the cells, followed by chloride ions (Cl ^- ), leads to cell swelling due to increased osmotic pressure, compression of neighboring neurons and their processes, which ultimately leads to lysis (cell rupture ) and the development of inflammatory reactions. Overall, this disruption of ion homeostasis leads to excitotoxicity, cell swelling and cell death, whereby damage spreads to adjacent tissues and pathological changes occur through secondary mechanisms. There is a need for effective treatment within the first 24 hours to protect stressed brain cells. The distribution of brain damage in stroke is similar to that seen in other forms of brain damage such as trauma and concussions.

Beyond acute treatment, efficient astrocyte function plays a key role in the broader restoration of neural tissue function—from 24 to 96 hours after a brain insult, over a period of months to years in patients with neurodegeneration such as Alzheimer's disease, or more generally case, in older people. Due to the inability of brain cells to regenerate, it is necessary to ensure that the remaining intact brain tissue is reorganized to restore any loss of function. This potential for neuronal reorganization decreases in older people.

G protein-coupled receptors (GPCRs) are thought to mediate cardioprotection. Therefore, there is the potential to treat heart disease and cardiovascular disease through similar mechanisms of action through modulation of these receptors.

There is a pressing and critical unmet medical need for more effective ways to treat brain injury, central nervous system injury, heart disease and cardiovascular disease and related conditions, and to promote recovery

- 1 046380 nervous tissue function in patients with neurodegenerative conditions such as Alzheimer's disease.

Brief description of the invention

It has now been discovered that the compounds of the present invention and their compositions can be used to treat, facilitate or accelerate recovery from certain conditions of the brain, central nervous system (CNS) or cardiovascular system, such as brain injury, neurodegenerative condition or ischemia hearts. In general, the free base forms and pharmaceutically acceptable compositions thereof are suitable for treating or reducing the severity of various diseases or disorders, as described in detail herein. Such compounds are represented by the chemical structure below, designated compound A:

The compounds of the present invention and pharmaceutically acceptable compositions thereof are suitable for the treatment of various diseases, disorders or conditions, including those described herein.

Brief description of graphic materials

In fig. Figure 1 shows the XRPD diffraction pattern of Form A of Compound A.

In fig. Figure 2 shows the DSC curve of Form A of Compound A.

In fig. Figure 3 shows the TGA curve of Form A of Compound A.

In fig. Figure 4 shows the chipboard curve of Form A of connection A.

In fig. Figure 5 shows an XRPD diffraction pattern of Form A of Compound A before (top) and after (bottom) DSP analysis.

In fig. 6 shows the IR-PF spectra of Form A of compound A.

In fig. Figure 7 shows the Raman-PF spectra of Form A of compound A.

In fig. Figure 8 shows the solid-state ¹³ C spectra of Form A of compound A.

In fig. Figure 9 is an optical microscope image of Form A of Compound A.

In fig. 10 shows the XRPD diffraction pattern of Form B of compound A.

In fig. Figure 11 shows the DSC curve of Form B of compound A.

In fig. 12 shows the TGA curve of Form B of Compound A.

In fig. 13 shows the chipboard curve of Form B of connection A.

In fig. 14 shows an XRPD diffraction pattern of Form B of Compound A before (top) and after (bottom) DSP analysis.

In fig. 15 shows the IR-PF spectra of Form B of compound A.

In fig. 16 shows the Raman-PF spectra of Form B of compound A.

In fig. 17 shows solid-state ¹³ C spectra of Form B of compound A. FIG. 18 is an optical microscope image of Form B of Compound A.

Detailed Description of the Invention

General Description of Certain Aspects of the Present Invention

US Patent No. 9,789,131, filed April 21, 2011, issued October 17, 2017 (the '131 patent), the contents of which are hereby incorporated by reference in their entirety, and US Patent Application Serial No. 15/ 670,738, filed August 7, 2017 and published as US 2018/0021363 on January 25, 2018 (Pub. '363), the contents of which are hereby incorporated by reference in their entirety, disclose certain therapeutically useful compounds. Such compounds include compound A:

- 2 046380

A.

In the '131 patent, Compound A is designated MRS4322, and the synthesis of Compound A is described in detail in Example 9 of the '131 Patent and is reproduced herein for ease of reference. In the '363 publication, compound A is designated MRS4322, and the synthesis of compound A is described in detail in Example 9 of the '363 publication and is reproduced herein for ease of reference.

It would be desirable to provide a solid form of Compound A (eg, its free base) that provides characteristics such as improved water solubility, stability, and ease of formulation. Accordingly, the present invention provides free base forms of Compound A.

Free base forms of compound A

It goes without saying that compound A can exist in various physical forms. For example, compound A may be in solution, suspension or solid form. In some embodiments, Compound A is in solid form. When compound A is in solid form, said compound may be amorphous, crystalline, or a mixture thereof. Examples of solid forms are described in more detail below.

In some embodiments, the present invention provides a form of Compound A that is substantially free of impurities. As used herein, the term substantially free of impurities means that the compound does not contain a significant amount of foreign matter. Such foreign matter may include various forms of Compound A, residual solvents, or any other impurities that may result from the preparation and/or isolation of Compound A. In some embodiments, the content of the form of Compound A is at least about 95% by weight. In other embodiments of the present invention, the content of the form of Compound A is at least about 99% by weight.

In one embodiment, Compound A is present in an amount of at least about 97, 97.5, 98.0, 98.5, 99, 99.5, 99.8 percent by weight, where the percentages are based on total mass of the composition. In another embodiment, Form A contains no more than about 3.0 HPLC area % of total organic impurities and, in some embodiments, no more than about 1.5 HPLC area % of total organic impurities relative to the total area of the HPLC chromatogram . In other embodiments, Compound A form contains no more than about 1.0% of the area of the HPLC chromatogram of any individual impurity; no more than about 0.6% of the area of the HPLC chromatogram of any individual impurity, and in some embodiments, no more than about 0.5% of the area of the HPLC chromatogram of any individual impurity relative to the total area of the HPLC chromatogram.

It is also intended that the structure depicted for the form of compound A include all tautomeric forms of compound A. In addition, the structures depicted herein are intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structure, except for the replacement of a hydrogen atom by a deuterium or tritium atom or the replacement of a carbon atom by a carbon atom enriched with ¹¹ C-, ¹³ C- or ¹⁴ C, are within the scope of the present invention.

It has been discovered that compound A can exist in various solid forms. Examples of such forms include polymorphic forms, such as the polymorphic forms described herein.

As used herein, the term polymorphic form refers to the various crystal structures into which a compound or a salt or solvate thereof may crystallize.

In some embodiments, Compound A is a crystalline solid. In other embodiments, compound A is a crystalline solid substantially free of amorphous compound A. As used herein, the term substantially free of amorphous compound A means that the compound does not contain a significant amount of amorphous compound A. In some embodiments, the content of crystalline compound A is at least about 95% by weight. In other embodiments of the present invention, the content of crystalline Compound A is at least about 99% by weight. It was discovered that compound A could exist for at least

- 3 046380 pe in two different polymorphic forms. Some embodiments of the present invention provide a polymorphic form of Compound A, referred to herein as Form A. Some embodiments of the present invention provide a polymorphic form of Compound A, referred to herein as Form B.

In some embodiments, Compound A is amorphous. In some embodiments, Compound A is an amorphous substance and is substantially free of crystalline Compound A.

Form A of Compound A

In some embodiments, Form A of Compound A has at least 1, 2, 3, 4, or 5 spectral peaks selected from the peaks listed in Table. 1 below.

Table 1 ______XRPD peak positions for Form A of compound A______

θΣΘ ¹ Relative intensity °20 Relative intensity °20 Relative intensity 7.6 1.49 21.5 35.63 30.8 12.79 8.0 100 22.8 2.30 32.0 8.32 9.0 3.49 23.1 6.16 32.8 8.27 10.8 4.60 23.7 4.21 32.9 13.11 11.8 1.59 23.9 9.39 33.7 9.73 12.5 4.59 24.9 54.39 34.7 4.31 13.1 59.02 26.1 32.89 36.3 3.98 16.2 34.56 26.5 13.26 36.7 8.82 16.7 37.68 26.6 22.22 37.9 12.68 17.2 10.65 27.1 60.62 38.2 3.16 17.9 45.59 28.6 10.52 38.5 1.75 18.1 16.05 29.3 1.66 38.7 2.83 18.3 10.91 29.7 9.05 39.6 2.37 19.8 4.96 30.1 1.89 — — 21.0 30.62 30.4 1.77 — —

*In this and all subsequent tables, position 26 is within ± 0.2.

In some embodiments, Form A of Compound A is characterized by having one or more peaks in its X-ray powder diffraction (XRPD) pattern selected from peaks at about 8.0 and about 13.1 degrees 2-theta. In some embodiments, Form A of Compound A is characterized by having two peaks in its X-ray powder diffraction pattern selected from peaks at about 8.0 and about 13.1 degrees 2-theta. As used herein, the term approximately when used in relation to a degree 2-theta value refers to the specified value of ±0.2 degrees 2-theta.

In some embodiments, the X-ray powder diffraction pattern is substantially similar to the XRPD shown in FIG. 1.

Methods for preparing Form A of Compound A are described below.

Form B of compound A

In some embodiments, Form B of Compound A has at least 1, 2, 3, 4, or 5 spectral peaks selected from those listed in Table. 2 below.

- 4 046380

table 2

XRPD peak positions for Form B of Compound A

θΣΘ ¹ Relative intensity °20 Relative intensity °20 Relative intensity 4.7 2.43 20.1 88.35 29.5 4.27 7.6 3.96 21.0 42.70 29.9 3.19 9.5 14.73 21.5 68.69 30.2 6.20 10.0 10.55 23.0 2.13 30.6 7.95 10.5 7.20 23.8 53.95 31.6 4.20 13.8 20.73 24.3 8.09 32.3 1.42 14.2 10.53 24.6 3.80 32.7 5.03 14.7 40.39 25.4 5.74 33.1 4.67 15.2 6.59 25.6 8.51 33.6 3.56 15.4 14.70 25.9 35.31 35.9 7,I 16.2 4.80 26.2 20.76 37.0 2.87 17.1 24.58 26.6 16.53 37.4 1.78 17.9 58.03 27.6 7.29 39.0 1.26 18.3 12.94 28.8 25.11 — — 19.0 100 29.1 8.40 — —

*In this and all subsequent tables, the position of 2Θ is within ± 0.2.

In some embodiments, Form B of Compound A is characterized by one or more peaks in its X-ray powder diffraction pattern selected from peaks at about 19.0, about 20.1, and about 21.5 degrees 2-theta. In some embodiments, Form B of Compound A is characterized by two or more peaks in its X-ray powder diffraction pattern selected from peaks at about 19.0, about 20.1, and about 21.5 degrees 2-theta. In some embodiments, Form B of Compound A is characterized by all three peaks in its X-ray powder diffraction pattern selected from peaks at about 19.0, about 20.1, and about 21.5 degrees 2-theta.

In some embodiments, Form B of Compound A is characterized by one or more peaks in its X-ray powder diffraction pattern selected from peaks at about 9.5, about 10.5, and about 13.8 degrees 2-theta. In some embodiments, Form B of Compound A is characterized by two or more peaks in its X-ray powder diffraction pattern selected from peaks at about 9.5, about 10.5, and about 13.8 degrees 2-theta. In some embodiments, Form B of Compound A is characterized by three peaks in its X-ray powder diffraction pattern selected from peaks at about 9.5, about 10.5, and about 13.8 degrees 2-theta.

In some embodiments, the X-ray powder diffraction pattern substantially matches the XRPD shown in FIG. 10. Methods for preparing Form B of Compound A are described below.

In some embodiments, the present invention provides Compound A:

wherein said compound is crystalline.

Some embodiments of the present invention provide a solid form of Compound A, wherein said compound is substantially free of amorphous Compound A.

In some embodiments, the present invention provides a solid form of Compound A, wherein the compound is substantially free of impurities. In some implementations

- 5046380 The present invention provides a solid form of compound A, wherein the compound is characterized by one or more peaks in its XRPD diffractogram selected from peaks at about 8.0 and about 13.1 degrees 2-theta. In some such embodiments, the present invention provides Compound 1, wherein the compound has two peaks in its XRPD diffractogram selected from peaks at about 8.0 and about 13.1 degrees 2-theta. Some such embodiments of the present invention provide Compound A, wherein said compound is Form A.

In some embodiments, the present invention provides a solid form of Compound A, wherein the compound has an XRPD diffraction pattern substantially consistent with the diffraction pattern depicted in FIG. 1.

Some embodiments of the present invention provide a solid form of Compound A, wherein the compound has one or more peaks in its XRPD diffractogram selected from peaks at about 19.0, about 20.1, and about 21.5 degrees 2-theta. Some such embodiments of the present invention provide Compound A, wherein the compound has at least two peaks in its XRPD diffractogram selected from peaks at about 19.0, about 20.1, and about 21.5 degrees 2-theta. Some embodiments of the present invention provide a solid form of Compound A, wherein the compound has one or more peaks in its XRPD diffractogram selected from peaks at about 9.5, about 10.5, and about 13.8 degrees 2-theta. Some such embodiments of the present invention provide Compound A, wherein the compound has at least two peaks in its XRPD diffractogram selected from peaks at about 0.5, about 10.5, and about 13.8 degrees 2-theta.

Some such embodiments of the present invention provide Compound A, wherein said compound is Form B.

Some embodiments of the present invention provide a solid form of Compound A, wherein the compound has an XRPD diffraction pattern substantially similar to that shown in FIG. 10.

In some embodiments, the present invention provides a composition comprising a solid form of Compound A and a pharmaceutically acceptable carrier or excipient.

In some embodiments, the present invention provides a compound selected from: Compound A in Form A and Compound A in Form B.

In some embodiments, the present invention provides a method of inhibiting or preventing the accumulation of cAMP in a patient, comprising administering to the patient a solid form of Compound A or a pharmaceutically acceptable composition containing the same.

In some embodiments, the present invention provides a method of treating an injury, disease or condition selected from traumatic brain injury (TBI), concussion, stroke, partial or complete spinal cord transection, malnutrition, toxic neuropathies, meningoencephalopathies, neurodegeneration caused by an inherited disease , age-related neurodegeneration, vascular disease, Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS), chronic traumatic encephalopathy (CTE), cardiovascular disease, autoimmune diseases, allergic diseases, transplant rejection, graft-versus-host disease, intraocular hypertension, glaucoma, odor sensitivity disorders, olfactory disorders, type 2 diabetes and/or pain management, respiratory diseases, central nervous system dysfunction, learning disabilities, cognitive impairment activity, hearing impairment, Meniere's disease, endolymphatic hydrops, progressive hearing loss, dizziness, vertigo, tinnitus, associated brain damage associated with radiation therapy for cancer, migraine treatment, sleep disorders in the elderly, epilepsy, schizophrenia, symptoms occurring in convalescents alcoholism, damage to neurons or nerves of the peripheral nervous system during surgery, gastrointestinal disorders, pain mediated by the central nervous system, migraine, concomitant brain damage associated with radiation therapy for cancer, depression, changes in mood or behavior, dementia, erratic behavior, suicidality , tremor, Huntington's chorea, decreased motor coordination, hearing loss, speech disorders, dry eye syndrome, hypomia, attention deficit, memory loss, cognitive impairment, vertigo, dysarthria, dysphagia, visual disturbances or disorientation, or addiction; comprising administering to the patient a solid form of Compound A or a pharmaceutically acceptable composition containing it. In some embodiments, the present invention provides a method of treating an injury, disease, or condition selected from traumatic brain injury (TBI), stroke, neurodegenerative condition, or heart disease or cardiovascular disease, comprising administering to a patient in need thereof an effective amount of an effective amount adenosine A ₃ receptor (A3R) agonist. In some embodiments, the adenosine A ₃ receptor (A ₃ R) agonist is a solid form of Compound A or a pharmaceutically acceptable component thereof.

- 6 046380 position. In some embodiments, the adenosine A3 receptor (A3R) agonist is a solid form of Compound A or a pharmaceutically acceptable composition thereof. In some embodiments, the solid form of Compound A or a pharmaceutically acceptable composition containing it acts through dual agonism at the adenosine A3 receptor and the adenosine A1 receptor (A1R).

In some embodiments, the present invention provides a method of treating an injury, disease, or condition selected from traumatic brain injury (TBI), stroke, neurodegenerative disease, heart disease, or cardiovascular disease, comprising administering to a patient in need thereof an effective amount of a dual agonist adenosine receptor (A3R), activity-biased A3R agonist, or activity-biased partial A3R agonist. In some embodiments, the adenosine A3 receptor (A3R) agonist is a solid form of Compound A or a pharmaceutically acceptable composition containing it. In some embodiments, the solid form of Compound A or a pharmaceutically acceptable composition containing it acts through dual agonism at the adenosine A3R receptor and the adenosine A1R receptor (A1R).

In some embodiments, the present invention provides a method of treating a brain or central nervous system (CNS) injury or condition selected from traumatic brain injury (TBI) or stroke, comprising administering to a patient in need thereof an effective amount of a solid form of Compound A or a pharmaceutically acceptable composition containing the same. . In some embodiments, the present invention provides a method of treating or alleviating traumatic brain injury (TBI), radiation injury, stroke, migraine headache, heart disease or cardiovascular disease, or neurodegenerative disorder, comprising administering to a patient in need thereof an effective amount of solid form of compound A or a pharmaceutically acceptable composition containing it.

In some embodiments, the present invention provides a method of treating or alleviating traumatic brain injury (TBI), radiation injury, stroke, migraine headache, heart disease or cardiovascular disease, or neurodegenerative disorder, comprising administering to a patient in need thereof an effective amount of solid form of compound A or a pharmaceutically acceptable composition containing it.

In some embodiments, the present invention provides a method of treating an injury, disease or condition selected from traumatic brain injury (TBI), stroke, neurodegenerative condition, or heart disease or cardiovascular disease, comprising administering to a patient in need thereof an effective amount of a solid form compound A or a pharmaceutically acceptable composition containing it.

In some embodiments, the injury, disease, or condition is a TBI.

In some embodiments, the TBI is selected from concussion, blast injury, combat trauma, or mild, moderate, or severe blow to the head.

In some embodiments, the injury, disease, or condition is a stroke selected from ischemic stroke, hemorrhagic stroke, subarachnoid hemorrhage, cerebral vasospasm, or transient ischemic attack (TIA).

In some embodiments, the patient experiences increased neuroprotection or recovery of neural tissue function compared to that of a patient not receiving treatment.

In some embodiments, the neurodegenerative disease is selected from Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS), chronic traumatic encephalopathy (CTE), or a neurodegenerative condition caused by a virus, alcoholism, tumor, toxin, or repeated brain damage.

In some embodiments, the neurodegenerative disease is Parkinson's disease.

In some embodiments, the injury, disease, or condition is Alzheimer's disease, migraine, brain surgery, or a neurological side effect due to cancer chemotherapy. In some embodiments, the recovery period from TBI, stroke, cardiac ischemia, or myocardial infarction is reduced compared to that of an untreated patient.

In some embodiments, the heart disease or cardiovascular disease is selected from cardiac ischemia, myocardial infarction, cardiomyopathy, coronary artery disease, arrhythmia, myocarditis, pericarditis, angina, hypertensive heart disease, endocarditis, rheumatic heart disease, congenital heart disease, or atherosclerosis.

In some embodiments, the heart disease or cardiovascular disease is cardiac ischemia or myocardial infarction.

In some embodiments, the compound or composition is administered over a long period of time

- 7 046380 tions for the treatment of stroke, cardiac ischemia or myocardial infarction during the period of time from the onset of damage to its elimination.

In some embodiments, the present invention provides a method of enhancing neuroprotection or restoring neural tissue function in a TBI or stroke patient in need thereof, comprising administering to said patient an effective amount of a solid form of Compound A or a pharmaceutically acceptable composition containing the same.

In some embodiments, the compound or a pharmaceutically acceptable salt thereof is administered orally, intravenously, or parenterally.

In some embodiments, the compound or composition is administered within 24 hours of TBI or stroke.

In some embodiments, the compound or composition is administered within 8 hours of TBI or stroke.

In some embodiments, the compound or composition is administered for at least the first 8 to 48 hours after TBI or stroke.

In some embodiments, the present invention provides a method of treating a heart disease or cardiovascular disease, comprising administering to a patient in need thereof an effective amount of a solid form of Compound A or a pharmaceutically acceptable composition containing the same.

In some embodiments, the patient has suffered cardiac ischemia or myocardial infarction.

In some embodiments, the compound or composition provides enhanced cardioprotection or regeneration of damaged heart tissue in a patient.

In some embodiments, the compound or composition provides a shorter recovery period from cardiac ischemia or myocardial infarction in a patient compared to that of an untreated patient.

In some embodiments, the present invention provides a method of treating an injury, disease, disorder, or condition selected from:

(i) radiation-induced brain damage or concomitant brain damage associated with radiation therapy for cancer or treatment for migraines;

(ii) migraine headache;

(iii) a condition associated with brain damage or a neurodegenerative condition; or (iv) an autoimmune disease or condition, glaucoma, hearing impairment, progressive hearing loss, tinnitus, epilepsy, analgesia, CNS-mediated pain, neuropathic pain, inflammatory pain or acute pain; comprising administering to a patient in need thereof an effective amount of a solid form of Compound A or a pharmaceutically acceptable composition containing the same. In some embodiments, the compound or composition provides increased neuroprotection or restoration of neural tissue function in a patient compared to that of an untreated patient.

In some embodiments, the condition associated with brain damage or a neurodegenerative condition is selected from epilepsy, migraine, concomitant brain injury associated with radiation therapy for cancer, depression, mood or behavioral changes, dementia, erratic behavior, suicidality, tremor, chorea Huntington's, decreased motor coordination, hearing loss, speech disorders, dry eye syndrome, hypomimia, attention deficit, memory loss, cognitive impairment or failure of cognitive performance, central nervous system dysfunction, learning disabilities, vertigo, dysarthria, dysphagia, visual disturbances or disorientation . In some embodiments, the present invention provides a method of enhancing cardioprotection or regeneration of damaged cardiac tissue in a patient in need thereof who has suffered cardiac ischemia or myocardial infarction, comprising administering to the patient an effective amount of a solid form of Compound A or a pharmaceutically acceptable composition containing the same. In some embodiments, the present invention provides a method for preparing a solid form of Compound A, comprising one or more steps of solvent removal and solvent addition. In some embodiments, the added solvent is the same as the removed solvent. In some embodiments, the added solvent is different from the removed solvent. Solvent removal agents are known in the art of synthesis and chemistry and include, but are not limited to, any of the agents described herein and in the Examples section.

In some embodiments, a method for preparing a solid form of Compound A includes one or more steps of heating or cooling the formulation.

In some embodiments, a method for preparing a solid form of Compound A includes one or more steps of mixing the drug by shaking or vortexing.

In some embodiments, a method for preparing a solid form of Compound A includes the step of heating.

- 8 046380

In some embodiments, a solid form of Compound A is precipitated from the mixture. In another embodiment, the solid form of Compound A is crystallized from the mixture. In other embodiments, the solid form of Compound A is crystallized from solution after seeding the solution (ie, adding crystals of Compound A to the solution).

The solid form of Compound A may precipitate from the reaction mixture or may be prepared by removing part or all of the solvent through methods such as evaporation, distillation, filtration (e.g., nanofiltration, ultrafiltration), reverse osmosis, absorption and reaction, by adding antisolvents, such as heptane, by cooling or by various combinations of these methods.

As generally described above, a solid form of Compound A may optionally be isolated. It should be understood that the solid form of compound A can be isolated using any suitable physical means known to one skilled in the art. In some embodiments, the precipitated solid form of Compound A is separated from the supernatant by filtration. In other embodiments, the precipitated solid form of Compound A is separated from the supernatant by decanting the supernatant.

In some embodiments, the solid form of Compound A is separated from the supernatant by filtration.

In some embodiments, the isolated solid form of Compound A is air dried. In other embodiments, the isolated solid form of Compound A is dried under reduced pressure, optionally at elevated temperature. Examples of suitable solvents suitable for use in the present invention include, but are not limited to, protic solvents, aprotic solvents, polar aprotic solvent, or mixtures thereof. In some embodiments, suitable solvents include an ether, an ester, an alcohol, a ketone, or mixtures thereof. In some embodiments, the solvent is one or more organic alcohols. In some embodiments, the solvent is chlorinated. In some embodiments, the solvent is an aromatic solvent.

In some embodiments, a suitable solvent is methanol, ethanol, isopropanol, or acetone, where the solvent is anhydrous or in combination with water or heptane. In some embodiments, suitable solvents include tetrahydrofuran, dimethylformamide, dimethyl sulfoxide, glyme, diglyme, methyl t-butyl ether, t-butanol, n-butanol, and acetonitrile. In some embodiments, a suitable solvent is ethanol. In some embodiments, a suitable solvent is anhydrous ethanol. In some embodiments, a suitable solvent is MTBE (methyl tert-butyl ether). In some embodiments, a suitable solvent is ethyl acetate. In some embodiments, the suitable solvent is a mixture of methanol and methylene chloride. In some embodiments, a suitable solvent is a mixture of acetonitrile and water. In some embodiments, a suitable solvent is methyl acetate, isopropyl acetate, acetone, or tetrahydrofuran. In some embodiments, a suitable solvent is diethyl ether. In some embodiments, a suitable solvent is water. In some embodiments, a suitable solvent is methyl ethyl ketone. In some embodiments, a suitable solvent is toluene.

Uses of the Compounds and Pharmaceutically Acceptable Compositions Thereof As used herein, the terms therapy, treat and treatment refer to reversing, alleviating, delaying the onset or slowing down the progression of a disease or disorder or one or more symptoms thereof as described herein. In some embodiments, therapy is administered after the development of one or more symptoms. In other embodiments, therapy is administered in the absence of symptoms. For example, therapy is administered to a susceptible individual prior to the onset of symptoms (eg, in light of symptom history and/or in light of genetic or other susceptibility factors). Therapy is also continued after symptoms have resolved, for example to prevent, delay or reduce the severity of their recurrence.

Injuries and conditions of the brain, central nervous system, cardiovascular system and other injuries and conditions

In some embodiments, the present invention provides a new approach for preventing and/or treating brain damage associated with acute traumatic brain injury, as well as chronic brain and central nervous system diseases and heart diseases and cardiovascular diseases and conditions. In one aspect, the present invention provides methods for treating such injuries, diseases and conditions by utilizing the neuroprotective and functional restoration effects of neural tissue mediated by astrocytes, which are now considered to be the key natural guardian cells of neurons, as well as astrocyte mitochondria, which provide a significant portion of energy needs brain. In another aspect, the present invention provides methods for treating such injuries, diseases and conditions from to

- 9 046380 the power of cardioprotective and regenerative effects mediated by A3R receptors. With regard to the effects of neuroprotection and restoration of function of nervous tissue, it is believed, without being bound by theory, that selective enhancement of astrocyte energy metabolism, mediated by A3R and/or P2Y1 receptors, stimulates the functions of astrocytes as guardian cells, for example, their functions providing neuroprotection and restoration of function nervous tissue, which in turn increases the resistance of neurons and other cells to both acute damage and chronic stress. In some cases, it may be appropriate to provide biased, that is, selective or preferential, activation of one or more signal transduction pathways mediated by the A3R and/or P2Y1 and/or A1R receptors, when activation of one or more undesired signaling pathways does not occur, or occurs to a lesser extent. degrees. In addition to, or as an alternative to, astrocytes, functions providing neuroprotection or restoration of neural tissue function may be activated in glia, microglia, neurons, endothelial cells, and other cell types in the brain and/or CNS. Accordingly, in one aspect of the present invention, compounds and methods of using them are provided for treating, facilitating or accelerating recovery from certain conditions of the brain or central nervous system (CNS), such as brain injury, for example, providing enhanced neuroprotective and/or functional restoration effects nervous tissue mediated by astrocytes, glia, microglia, neurons, endothelial cells or other cells of the brain and/or central nervous system, comprising administering to a patient in need thereof an effective amount of a solid form of Compound A or a pharmaceutically acceptable composition containing the same.

Astrocytes play key roles in the maintenance and protection of neurons, and they significantly influence the effects of traumatic brain injury, such as ischemic injury. Less appreciated is the central role that astrocyte mitochondria themselves play in these brain functions. For example, inhibition of astrocyte mitochondrial function increases cell swelling and leads to necrotic cell death. Neurons undergo permanent damage due to repeated propagating depolarizations only if astrocyte mitochondrial function is impaired, and astrocyte mitochondria are required to reduce the pathophysiological increase in extracellular K' ions. which initiates propagating depolarizations. Activation of purinergic receptors on astrocytes leads to an increase in the content of mitochondrial Ca ²⁺ ions, which enhances the function of the citric acid cycle in mitochondria and increases the intensity of respiration and ATP production. Accordingly, in one aspect, the present invention relates to the discovery that activation of astrocyte purinergic receptors enhances the function of signaling pathways mediating brain cell survival, thereby ensuring the viability of both astrocytes and neurons under oxidative stress. In addition, activated astrocytes produce and deliver reduced glutathione, a key antioxidant that promotes the resistance of both astrocytes and neurons to oxidative stress. Thus, in one aspect of the present invention, there is provided a method of modulating astrocyte purinergic receptors to promote the survival and viability of one or more cell types in the brain of a patient following oxidative stress, such as oxidative stress caused by brain injury, ischemia reperfusion, or a neurodegenerative condition, comprising administering to the patient in need thereof, a solid form of Compound A or a pharmaceutically acceptable composition containing it.

In some embodiments, activation of astrocytes is achieved by bringing one or more purinergic receptors, such as adenosine receptors (AR), for example, associated with or expressed by astrocytes, into contact with an exposed compound, thereby modulating the activity of one or more receptors. In some embodiments, the compound activates astrocytes by acting on adenosine receptors on astrocytes, such as A1, A2A, A2b, and A3 receptors, to treat one or more disclosed diseases or conditions. In some embodiments, the disclosed compound, when administered to a patient in need thereof, affects one or more functions such as glutamate uptake, reactive gliosis, cell swelling, and the release of neurotrophic and neurotoxic factors affecting metabolic stress and its consequences, and thus thus providing treatment for one or more diseases or conditions. In some embodiments, the compound is an AR agonist. In some embodiments, the purinergic receptor is an adenosine A ₃ receptor (A3R). In some embodiments, the compound is an A3R agonist. In some embodiments, the compound is an A3 receptor (A3R) partial agonist, such as the human A3 receptor (hA3R), or a potency-biased A3R agonist, or a potency-biased partial A3R agonist. In some embodiments, the compound is a potency-biased A3 receptor antagonist. In some embodiments, the compound acts through dual agonism at A3R and A1R. In some embodiments, the compound is a solid form of Compound A or a pharmaceutically acceptable composition thereof.

- 10 046380

P2Y receptors are G protein-coupled receptors, and various subtypes of these receptors play important roles in processes such as synaptic communication, cell differentiation, ion flow, vasodilation, blood-brain barrier permeability, platelet aggregation, and neuromodulation. Characteristic members of the P2Y purinergic receptor family include mammalian receptors P2Y1, P2Y11, P2Y ₁₂ , and P2Y13, which bind adenine nucleotides; receptors P2Y ₄ , P2Y ₆ and P2Y ₁₄ , which bind to uracil nucleotides; and rodent P2Y2 receptors and P2Y4 receptors, which have mixed selectivity. In some embodiments, activation of astrocytes is achieved by bringing one or more purinergic receptors, such as P2Y receptors, such as those associated with or expressed by astrocytes, into contact with an exposed compound, thereby modulating the activity of the one or more receptors. In some embodiments, the compound activates astrocytes by targeting P2Y receptors, such as P2Y1, P2Y11, P2Y12, and P2Y13 receptors associated with or expressed by astrocytes, to treat one or more disclosed diseases or conditions. In some embodiments, the P2Y receptor is a P2Y| receptor. In some embodiments, the P2Y receptor _| located on intracellular mitochondrial membranes. In some embodiments, the compound is a P2Y agonist. In some embodiments, the compound is a P2Y agonist _| eg P2Y receptor agonist _| person. In some embodiments, the compound is a P2Y receptor agonist _| , such as the P2Y receptor _| human, with shifted activity, partial P2Y agonist _| or partial P2Y agonist _| with shifted activity. In some embodiments, the compound is a P2Y|receptor antagonist. with shifted activity. In some embodiments, the compound is a solid form of Compound A or a pharmaceutically acceptable composition thereof.

In another aspect, the present invention provides a method of treating or alleviating brain damage, such as brain damage due to TBI or a progressive neurodegenerative disorder, in a patient in need thereof, comprising administering to the patient an effective amount of a disclosed compound. In some embodiments, the subject has suffered a TBI, concussion, stroke, partial or complete spinal cord transection, or suffered from poor nutrition. In other embodiments, the subject suffers from toxic neuropathies, meningoencephalopathies, neurodegeneration caused by an inherited disease, age-related neurodegeneration, or vascular disease; or other disease disclosed in US Pat. No. 8,691,775, which is incorporated herein by reference. In some embodiments, the present invention provides a method of treating or alleviating brain damage, such as brain damage due to TBI or a progressive neurodegenerative disorder, in a patient in need thereof, comprising administering to the patient an effective amount of an A3R agonist. In other embodiments, the present invention provides a method of treating or alleviating brain damage, such as brain damage due to TBI or a progressive neurodegenerative disorder, in a patient in need thereof, comprising administering to the patient an effective amount of a P2Y ₁ agonist. In some embodiments, the compound is a potency-biased A ₃ receptor agonist, a partial A ₃ agonist, or a potency-biased partial A ₃ agonist. In some embodiments, the compound acts through dual agonism at A3R and A ₁ R. In some embodiments, the compound is a potency-biased P2Y1 receptor agonist, a partial P2Y ₁ agonist, or a potency-biased partial P2Y ₁ agonist, or a P2Y ₁ antagonist. In some embodiments, the compound is a solid form of Compound A or a pharmaceutically acceptable composition thereof. In another aspect of the present invention, there is provided a method of promoting or enhancing neuroprotection, restoring neural tissue function, or regenerating neural tissue in a patient suffering from a disease or condition, comprising administering to said patient an effective amount of a disclosed compound, for example, a solid form of Compound A or a pharmaceutically acceptable composition containing the same. . In some embodiments, the patient suffers from a neurodegenerative disease or condition. In some embodiments, the patient has suffered a TBI.

In another aspect, the present invention provides a method of promoting astrocyte-mediated neuroprotection or restoration of neural tissue function in a patient in need thereof, comprising administering to the patient an effective amount of a disclosed compound. In some embodiments, the present invention provides a method of promoting astrocyte-mediated neuroprotection or restoration of neural tissue function in a patient in need thereof, comprising administering to the patient an effective amount of an A3R agonist. In other embodiments, the present invention provides a method of promoting astrocyte-mediated neuroprotection or restoration of neural tissue function in a patient in need thereof, comprising administering to the patient an effective amount of a P2Y ₁ agonist. In some embodiments, the compound is an activity-biased A ₃ receptor agonist, a partial A ₃ agonist, or

- 11 046380 partial A3 agonist with shifted activity or A3 antagonist. In some embodiments, the compound acts through dual agonism at A3R and A1R. In some embodiments, the compound is an activity-biased P2Y ₁ receptor agonist, a partial P2Y|agonist. or a partial P2Y agonist, with biased activity, or a P2Yj antagonist. In some embodiments, the compound is a solid form of Compound A or a pharmaceutically acceptable composition thereof.

In another aspect of the present invention, there is provided a method of promoting the survival of neurons, glial cells, endothelial cells or other brain cells, such as cells in the ischemic penumbra, in a patient in need thereof, comprising administering to the patient an effective amount of a compound disclosed herein. In another aspect of the present invention, there is provided a method of promoting the survival of neurons, glial cells or other brain cells, such as cells in the ischemic penumbra, in a patient in need thereof, comprising administering to the patient an effective amount of an A3R agonist. In another aspect of the present invention, there is provided a method of promoting the survival of neurons, glial cells, endothelial cells or other brain cells, such as cells in the ischemic penumbra, in a patient in need thereof, comprising administering to the patient an effective amount of a P2Y ₁ agonist. In some embodiments, the compound is an activity-biased _A3 receptor agonist, an _A3 partial agonist, or an activity-biased partial _A3 agonist, or an _A3 antagonist. In some embodiments, the compound acts through dual agonism at A ₃ R and A ₁ R. In some embodiments, the compound is a potency-biased P2Y ₁ receptor agonist, a partial P2Y1 agonist, or a biased P2Y1 partial agonist, or a P2Y _v antagonist. In some embodiments, the compound is a solid form of Compound A or a pharmaceutically acceptable composition thereof. In additional embodiments, the patient suffers from or is at risk of suffering brain damage, such as those described below. Accordingly, methods for treating the conditions discussed below are also provided.

Traumatic brain injuries

Traumatic brain injury (TBI) is an unfortunately common condition and is projected to become the third leading cause of morbidity and mortality by 2020 worldwide. There are no approved treatments for TBI, and most patients with TBI are discharged from the hospital without pharmacological treatment (Witt 2006). Repeated TBI, such as concussions, can trigger age-related neurodegeneration, which leads to a variety of symptoms and disabilities over decades (McKee 2013). TBIs can occur as a result of sports injuries, motor vehicle accidents, falls, explosions, personal injury, and the like. Injuries range widely in complexity and severity from mild concussions with short-term changes in mental status, cognitive impairment, or loss of consciousness to severe unconsciousness and/or amnesia for long periods of time after injury. In the United States, approximately 1.7 million people annually sustain a TBI and seek medical treatment (according to USCSF and CDC), and the CDC estimates that an additional 1.6 to 3.8 million concussions occur each year. during sports and other recreational activities where victims do not go to hospitals or emergency rooms. (CDC; Langlois 2006) Approximately 5-10% of athletes suffer a concussion during each sporting season. (Sports Concussion Institute 2012) American football is the sport associated with the highest risk of concussion for men (75% chance of concussion), while European football has the highest risk of concussion for women (50% chance of concussion). TBI is the leading cause of death and disability among children and young adults (CDC) and the most commonly sustained injury in military operations; approximately 20% of US military personnel mobilized since 2003 have suffered at least one TBI. (Chronic Effects of Neurotrauma Consortium (CENC); Warden 2006; Scholten 2012; Taylor 2012; Gavett 2011; Guskiewicz 2005; Omalu 2005) Total indirect and direct medical costs associated with TBI are estimated at $77 billion per year (UCSF and CDC). At least 5 million Americans require ongoing daily assistance with activities due to TBI (CDC and Thurman 1999).

Activation of astrocytes in accordance with the present invention represents a new possible method for treating such conditions. Accordingly, one aspect of the present disclosure provides a method of treating TBI or promoting recovery from TBI, comprising administering to a patient in need thereof an effective amount of a disclosed compound. In some embodiments, the TBI is selected from traumatic injuries to the brain (such as concussion, blast injury, combat trauma) or the spinal cord (such as partial or complete transection of the spinal cord). In some embodiments, TBI results from a mild, moderate, or severe blow to the head, involves an open or closed head wound, or is the result of

- 12 046380 volume of a penetrating or non-penetrating blow to the head. In some embodiments, the present invention provides a method of treating TBI or promoting recovery from TBI, comprising administering to a patient in need thereof an effective amount of an A3R agonist. In some embodiments, the present invention provides a method of treating TBI or promoting recovery from TBI, comprising administering to a patient in need thereof an effective amount of a P2Y1 agonist. In some embodiments, the compound is a potency-biased A3 receptor agonist, an A3 partial agonist, or a potency-biased partial A3 agonist, or an A3 antagonist. In some embodiments, the compound acts through dual agonism at A3R and A1R. In some embodiments, the compound is a potency-biased P2Y1 receptor agonist, a partial P2Y1 agonist, or a potency-biased P2Y1 partial agonist, or a P2Y 1 antagonist. In some embodiments, the compound is a solid form of Compound A or a pharmaceutically acceptable composition thereof.

Stroke

A stroke occurs when the blood vessel that carries oxygen and nutrients to the brain is damaged due to ischemic blockage or hemorrhagic rupture of a blood vessel in the brain, causing the death of neurons, glia, and endothelial cells in the damaged area of the brain. The outcome of a stroke depends on the location and area of damage, and the effects of this damage are observed in the body functions regulated by the damaged area of the brain. Strokes can cause unilateral or bilateral paralysis, speech impairment and difficulty, memory loss, behavioral changes, and even death. Stroke is the fourth leading cause of death in the United States and a leading cause of disability in adults. Every year ~800,000 people suffer a new or recurrent stroke. More than 2,000 Americans suffer a stroke every day, and more than 400 of these cases result in death. In 2010, stroke accounted for ~1 in every 19 deaths in the United States. An estimated 6.8 million Americans aged >20 years have had a stroke. (American Health Association (AHA) and Go 2014) As of 2010, direct and indirect medical costs associated with stroke were estimated at $36.5 billion annually. Within minutes of a stroke, the lack of blood flow will cause permanent damage to the central mass of brain tissue. Between this damaged central mass of tissue and normal brain tissue is an area of tissue known as the ischemic penumbra—tissue that is subject to gradually increasing stress due to decreased blood flow and some disruption of energy metabolism. During the first 24-48 hours after stroke, the stress experienced by neurons and glial cells in the ischemic penumbra leads to either some recovery or further cell death.

In one aspect of the present invention, there is provided a method of neuroprotective therapy for a patient who has suffered a stroke, comprising administering to the patient in need thereof an effective amount of a disclosed compound. In some embodiments, such therapy rescues as much of the ischemic penumbra as possible and/or limits further acute tissue damage and/or stimulates neuronal recovery. In another aspect, there is provided a method of treating stroke or promoting recovery from stroke, comprising administering to a patient in need thereof an effective amount of a disclosed compound. In another aspect, there is provided a method of promoting or enhancing neuroprotection, neural tissue regeneration, or restoration of neural tissue function in a patient who has suffered a stroke, comprising administering to said patient an effective amount of a disclosed compound. In another aspect, there is provided a method of treating stroke or promoting stroke recovery, comprising administering to a patient in need thereof an effective amount of an A3R agonist. In some embodiments, the present invention provides a method of treating stroke or promoting stroke recovery, comprising administering to a patient in need thereof an effective amount of a P2Y1 agonist. In some embodiments, the compound is a potency-biased A3 receptor agonist, an A3 partial agonist, or a potency-biased partial A3 agonist, or an A3 antagonist. In some embodiments, the compound acts through dual agonism at A3R and A1R. In some embodiments, the compound is a potency-biased P2Y ₁ receptor agonist, a partial P2Y ₁ agonist, or a biased P2Y ₁ partial agonist, or a P2Y 1 antagonist. In some embodiments, the compound is a solid form of Compound A or a pharmaceutically acceptable composition thereof.

In some embodiments, the stroke is selected from ischemic stroke, hemorrhagic stroke, subarachnoid hemorrhage, cerebral vasospasm, or transient ischemic attack (TIA). In some embodiments, the stroke is an ischemic stroke. In some embodiments, the stroke is a hemorrhagic stroke. In some embodiments, the compound is administered within 48 hours of the stroke. In some embodiments, the compound is administered within 24 hours of the stroke. In some embodiments, the compound is administered within 16 hours of the stroke. In some implementations

- 13 046380 tions the compound is administered within 8, 4, 2 or 1 hour after the stroke. In some embodiments, the compound is administered for at least the first 1 to 72 hours after the stroke. In some embodiments, the compound is administered for at least the first 8 to 52 hours after the stroke. In some embodiments, the compound is administered for at least the first 8 to 48 hours after the stroke. In some embodiments, the compound is administered for at least the first 2448 hours after the stroke. In some embodiments, the compound is administered chronically to treat a stroke after it has occurred. In some embodiments, the compound is chronically administered to treat transient ischemic attacks (TIAs).

In some embodiments, the compound is chronically administered to treat ischemic stroke, hemorrhagic stroke, subarachnoid hemorrhage, cerebral vasospasm, transient ischemic attack (TIA), or to treat a patient at increased risk of stroke, such as a patient who has had a stroke in the past. and who is at risk of recurrent stroke, for example, a patient over 40, 45, 50, 55, 60, 65, 70, 75 or 80 years of age.

In some embodiments, the compound is used to treat ischemia-reperfusion injury caused by stroke.

Non-degenerative diseases

Neurodegenerative diseases are incurable, progressive, and ultimately disabling syndromes that result from the progressive degeneration and/or death of neurons in the brain and spinal cord. Neurodegeneration leads to movement disorders (ataxias) and/or cognitive impairment (dementias) and includes a spectrum of diseases such as Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), multiple sclerosis (MS), amyotrophic lateral disease multiple sclerosis (ALS) and chronic traumatic encephalopathy (CTE). While many neurodegenerative diseases are primarily hereditary, other causes may include viruses, alcoholism, tumors or toxins, and, as is now clear, repeated brain damage.

Over time, neurons accumulate cellular damage as a consequence of the above factors, which is generally considered to be the cause of many neurodegenerative diseases associated with long-term cellular stress, such as Alzheimer's disease and Parkinson's disease, in older people. Dementias are the main consequences of neurodegenerative diseases, with AD accounting for approximately 60–70% of cases. (Kandale 2013) As discussed above, activation of mechanisms of neuroprotection and restoration of neural tissue function can alleviate the progression of one or more neurodegenerative diseases. Accordingly, in one aspect of the present invention there is provided a method of treating a neurodegenerative disease or promoting recovery from a neurodegenerative disease, comprising administering to a patient in need thereof an effective amount of a solid form of Compound A or a pharmaceutically acceptable composition containing the same. In one aspect of the present invention, there is provided a method of promoting neuroprotection or restoration of neural tissue function in a patient suffering from a neurodegenerative disease, comprising administering to said patient an effective amount of a disclosed compound. In some embodiments, there is provided a method of promoting neuroprotection or restoration of neural tissue function in a patient suffering from a neurodegenerative disease, comprising administering to the patient an effective amount of an A ₃ R agonist. In other embodiments, there is provided a method of promoting neuroprotection or restoration of neural tissue function in a patient suffering from a neurodegenerative disease, comprising administering to said patient an effective amount of a P2Y ₁ agonist. In some embodiments, the compound is a potency-biased A3 receptor agonist, an A3 partial agonist, or a potency-biased partial A3 agonist, or an A3 antagonist. In some embodiments, the compound acts through dual agonism at A3R and A1R. In some embodiments, the compound is an activity-biased P2Y ₁ receptor agonist, a partial P2Y|agonist. or a partial P2Y ₁ agonist with biased activity or a P2Y1 antagonist. In some embodiments, the compound is a solid form of Compound A or a pharmaceutically acceptable composition thereof.

Alzheimer's disease (AD)

An estimated 5.2 million Americans of all ages suffered from AD in 2014; 11% of the population aged 65 years and older suffer from asthma. (Alzheimer's Association) By 2050, the number of people with AD aged 65 years and older is projected to nearly triple to 13.8 million. In the United States, medical costs for patients with AD are approximately $214 billion per year; 70% of these costs are covered by Medicare and Medicaid. On current trends, these costs will rise to $1.2 trillion per year by 2050.

Activation of astrocytes and promotion of neuroprotection and restoration of nerve tissue function in accordance with the present invention represents a new possible method for the treatment of AD.

- 14 046380

Accordingly, one aspect of the present disclosure provides a method of treating AD or promoting neuroprotection or restoration of neural tissue function in a patient suffering from AD, comprising administering to said patient an effective amount of a compound disclosed herein. In some embodiments, the present invention provides a method of treating AD or promoting neuroprotection or restoration of neural tissue function in a patient suffering from AD, comprising administering to the patient an effective amount of an A3R agonist. In some embodiments, the present invention provides a method of treating AD or promoting neuroprotection or restoration of neural tissue function in a patient suffering from AD, comprising administering to the patient an effective amount of a P2Y1 agonist. In some embodiments, the compound is a potency-biased A3 receptor agonist, an A3 partial agonist, or a potency-biased partial A3 agonist, or an A3 antagonist. In some embodiments, the compound acts through dual agonism at A3R and A1R. In some embodiments, the compound is a potency-biased P2Y1 receptor agonist, a partial P2Y1 agonist, or a potency-biased P2Y1 partial agonist, or a P2Y 1 antagonist. In some embodiments, the compound is a solid form of Compound A or a pharmaceutically acceptable composition thereof.

Parkinson's disease (PD)

At least one million Americans live with PD, and approximately 60,000 Americans are newly diagnosed with PD each year, not counting the thousands of cases that go undiagnosed. (Parkinson's Disease Foundation) The total direct and indirect medical costs associated with PD, including therapeutic treatments, Social Security benefits, and lost income, are estimated to total nearly $25 billion annually in the United States. (Parkinson's Disease Foundation and Huse 2005)

Activation of neuroprotection and restoration of nerve tissue function in accordance with the present invention represents a new possible method for the treatment of PD. Accordingly, one aspect of the present disclosure provides a method of treating PD or promoting neuroprotection or restoration of neural function in a patient suffering from PD, comprising administering to said patient an effective amount of a disclosed compound. In some embodiments, the present invention provides a method of treating PD or promoting neuroprotection or restoration of neural tissue function in a patient suffering from PD, comprising administering to said patient an effective amount of an A ₃ R agonist.

In some embodiments, the present invention provides a method of treating PD or promoting neuroprotection or restoration of neural tissue function in a patient suffering from PD, comprising administering to the patient an effective amount of a P2Y1 agonist. In some embodiments, the compound is a potency-biased A3 receptor agonist, an A3 partial agonist, or a potency-biased partial A3 agonist, or an A3 antagonist. In some embodiments, the compound acts through dual agonism at A3R and A1R. In some embodiments, the compound is a biased P2Y1 receptor agonist, a partial _P2Y1 agonist, or a biased _P2Y1 partial agonist, or a P2Y1 antagonist. In some embodiments, the compound is a solid form of Compound A or a pharmaceutically acceptable composition thereof.

Multiple sclerosis (MS)

More than 400,000 people in the United States suffer from multiple sclerosis. In young adults, MS is the most common disease of the central nervous system. (Multiple Sclerosis Foundation) Astrocytes are able to reverse MS-induced destruction of myelin sheaths on nerve cells through their effects of restoring nerve tissue function and promoting the healing of CNS damage in patients with MS.

Thus, the activation of neuroprotection and restoration of function of nervous tissue in the CNS in accordance with the present invention represents a new possible method for the treatment of MS. Accordingly, in one aspect of the present disclosure there is provided a method of treating MS or promoting neuroprotection or restoration of neural function in a patient suffering from MS, comprising administering to said patient an effective amount of a disclosed compound. In some embodiments, the present invention provides a method of treating MS or promoting neuroprotection or restoration of neural tissue function in a patient suffering from MS, comprising administering to the patient an effective amount of an A3R agonist. In some embodiments, the present invention provides a method of treating MS or promoting neuroprotection or restoring neural tissue function in a patient suffering from MS, comprising administering to said patient an effective amount of a P2Y 1 agonist. In some embodiments, the compound is an activity-biased A3 receptor agonist, partial A3 agonist or partial A3 agonist with biased activity or A3 antagonist. In some embodiments, the compound acts through dual agonism at A ₃ R and A ₁ R. In some embodiments, the compound is a biased P2Y ₁ receptor agonist, a partial P2Y ₁ agonist, or a biased P2Y ₁ partial agonist or P2Y ₁ antagonist. In some embodiments, the compound is a solid form of Compound A or a pharmaceutically acceptable composition thereof.

Amyotrophic lateral sclerosis (ALS)/Lou Gehrig's disease

ALS is diagnosed in approximately 5,600 people each year in the United States; along with this, at least 30,000 Americans may suffer from this disease. (ALS Association) Activation of astrocytes may promote repair and repair of neurons and their connections in a patient with ALS.

Accordingly, one aspect of the present disclosure provides a method of treating ALS or promoting neuroprotection or restoration of neural function in a patient suffering from ALS, comprising administering to said patient an effective amount of a disclosed compound. Other embodiments also provide a method of promoting recovery and repair of neurons and their connections in a patient with ALS, comprising administering to the patient an effective amount of a compound disclosed herein. In some embodiments, the present invention provides a method of treating ALS or promoting neuroprotection or restoring neural tissue function in a patient suffering from ALS, comprising administering to the patient an effective amount of an A ₃ R agonist. In some embodiments, the present invention provides a method of treating ALS or promoting neuroprotection or restoring neural tissue function in a patient suffering from ALS, comprising administering to said patient an effective amount of a P2Y1 agonist. In some embodiments, the compound is a potency-biased A3 receptor agonist, a partial A3 agonist, or a potency-biased partial A3 agonist, or an A3 antagonist. In some embodiments, the compound acts through dual agonism at A3R and A1R. In some embodiments, the compound is a potency-biased P2Y1 receptor agonist, a partial P2Y1 agonist, or a potency-biased partial P2Y1 agonist, or a P2Y1 antagonist. In some embodiments, the compound is a solid form of Compound A or a pharmaceutically acceptable composition thereof.

Chronic traumatic encephalopathy (CTE)

CTE (a form of tauopathy) is a progressive neurodegenerative disease found in people who have suffered one or more (often multiple or repeated over time) severe blows to the head. CTE is most commonly diagnosed in professional athletes involved in American football, European soccer, hockey, professional wrestling, stunt racing, bull riding and rodeo, motocross and other contact sports who have suffered traumatic brain injury and/or repeated concussions. A subgroup of CTE patients suffer from chronic traumatic encephalomyopathy (CTEM), which is characterized by symptoms of motor neuron disease that resemble those of ALS. Progressive muscle weakness and disturbances in movement and gait are considered early signs of CTEM. Symptoms of the first stage of CTE include progressive attention deficits, confusion, dizziness and headaches. Stage two symptoms include memory loss, social instability, erratic behavior, and inability to judge. In stages three and four, patients suffer from progressive dementia, slow movements, tremors, hypomia, vertigo, speech impediments, hearing loss and suicidality, and may also suffer from dysarthria, dysphagia and visual disturbances such as ptosis.

Accordingly, one aspect of the present disclosure provides a method of treating or preventing CTE or promoting neuroprotection or restoring neural tissue function in a patient suffering from CTE, comprising administering to said patient an effective amount of a disclosed compound. Other embodiments also provide a method of promoting repair and repair of neurons and their connections in a patient with CTE, comprising administering to the patient an effective amount of a disclosed compound. In some embodiments, the compound provides treatment for one or more symptoms of stage one, stage two, stage three, or stage four of CTE. In some embodiments, the present invention provides a method of treating CTE or promoting neuroprotection or restoring neural tissue function in a patient suffering from CTE, comprising administering to the patient an effective amount of an A ₃ R agonist. In some embodiments, the present invention provides a method of treating CTE or promoting neuroprotection or restoring neural tissue function in a patient suffering from CTE, comprising administering to said patient an effective amount of a P2Y1 agonist. In some embodiments, the compound is an activity-biased A3 receptor agonist, an A3 partial agonist, or an activity-biased partial A3 agonist, or an A3 antagonist. In some embodiments, the compound acts through dual agonism at A3R and A1R. In some embodiments, the compound is a potency-biased P2Y1 receptor agonist, a partial P2Y ₁ agonist, or a potency-biased partial P2Y ₁ agonist, or a P2Y ₁ antagonist. In some embodiments, the connection is solid

- 16 046380 form of compound A or a pharmaceutically acceptable composition containing it.

At the microscopic level, pathology includes neuronal death, deposition of tau protein, deposition of DNA transactivation response element (TAR) protein 43 (TDP 43) as beta-amyloid, changes in cerebral white matter, and other abnormalities. Tau deposition involves the increasing presence of dense neurofibrillary tangles (NFTs), neurites, and glial tangles, which are composed of astrocytes and other glial cells. Thus, in some embodiments, the method provides for the treatment, promotion of elimination, or prevention of neuronal death, tau protein deposition, TAR DNA binding protein 43 (TDP 43) beta-amyloid deposition, changes in cerebral white matter, and other disorders associated with CTE.

Some embodiments of the present invention provide long-term administration of a compound disclosed herein, such as a potency-biased _A3R agonist, a partial _A3R agonist, or a biased _A3R partial agonist, or a dual _A3R and A1R agonist, or an activity-biased P2Y ₁ agonist, a partial P2Y ₁ agonist, or an activity-biased partial P2Y ₁ agonist, for the treatment of a neurodegenerative disease such as those discussed above and below.

Cardiovascular diseases

The disclosed compounds are also suitable for use in the treatment of various cardiovascular diseases and conditions. In some embodiments, the present invention provides a method of treating a heart disease (heart disease) or cardiovascular disease, such as cardiac ischemia, myocardial infarction, cardiomyopathy, coronary artery disease, arrhythmia, myocarditis, pericarditis, angina, hypertensive heart disease, endocarditis, rheumatic fever. heart disease, congenital heart disease or atherosclerosis, comprising administering to a patient in need thereof an effective amount of a disclosed compound, such as a solid form of Compound A or a pharmaceutically acceptable composition thereof. In some embodiments, the disclosed compound modulates ATP-dependent potassium channels, for example, through activity-biased agonism, partial agonism or activity-biased partial agonism at the A ₃ R receptor, or dual agonism at the A ₃ R and A1R.

In some embodiments, the heart disease or cardiovascular disease is cardiac ischemia or myocardial infarction. In some embodiments, the present invention provides a method of promoting or enhancing cardioprotection, cardiac function restoration, or cardiac regeneration in a patient suffering from a heart disease (heart disease or condition) or a cardiovascular disease or condition, comprising administering to said patient an effective amount of a disclosed compound, e.g. , a solid form of Compound A or a pharmaceutically acceptable composition containing it.

In some embodiments, the heart disease (heart disease) or cardiovascular disease from which the patient is suffering is cardiac ischemia, myocardial infarction, cardiomyopathy, coronary heart disease, arrhythmia, myocarditis, pericarditis, angina, hypertensive heart disease, endocarditis, rheumatic heart disease , congenital heart disease or atherosclerosis. In some embodiments, the disclosed compound modulates ATP-dependent potassium channels, for example, through activity-biased agonism, partial agonism or activity-biased partial agonism at the A ₃ R receptor, or dual agonism at the A ₃ R and A1R.

Other diseases

Compounds that modulate beneficial effects such as neuroprotection, for example by increasing astrocytic mitochondrial activity, also have potential for treating a number of other diseases. For example, due to the role of astrocytes in neuroprotection disclosed in the present invention, activation of astrocytes, for example, through modulation of the A ₃ R and/or P2Y ₁ receptor, may be suitable for use in the treatment of various diseases and conditions discussed below.

Accordingly, some embodiments of the present invention provide a method of treating neurodegeneration in a patient suffering from a disease or condition, comprising administering to said patient an effective amount of a disclosed compound, for example, a solid form of Compound A or a pharmaceutically acceptable composition containing the same.

In some embodiments, the present invention provides a method of promoting or enhancing neuroprotection, restoring neural function, or regenerating neural tissue in a patient suffering from a disease or condition, comprising administering to said patient an effective amount of a disclosed compound, e.g., a solid form of Compound A or a pharmaceutically acceptable form containing the same compositions.

In some embodiments, the disease or condition is selected from autoimmune diseases, allergic diseases and/or graft rejection and graft-versus-host disease (for information on the use of certain nucleoside and nucleotide compounds in the treatment of these conditions, see, for example, WO 2007/20018, which included in this description by

- 17 046380 via reference). In other embodiments, the disease or condition is selected from ocular hypertension and/or glaucoma (for information on the use of certain nucleoside and nucleotide compounds in the treatment of these conditions, see, for example, WO 2011/77435, which is incorporated herein by reference). In other embodiments, the disease or condition is selected from odor sensitivity and/or smell impairment (for information on the use of certain nucleoside and nucleotide compounds in the treatment of these conditions, see, for example, EP1624753, which is incorporated herein by reference). In other embodiments, the disease or condition is selected from type 2 diabetes and/or pain management (for information on the use of certain nucleoside and nucleotide compounds in the treatment of these conditions, see, for example, US 2010/0256086, which is incorporated herein by reference).

In other embodiments, the disease or condition is selected from respiratory diseases and/or cardiovascular (CV) diseases (for information on the use of certain nucleoside and nucleotide compounds in the treatment of these conditions, see, for example, FASEB J. (2013) 27: 1118.4 (meeting abstracts), which is incorporated herein by reference). In other embodiments, the disease or condition is selected from CNS dysfunction, learning disabilities, and/or cognitive deficits (for information on the use of certain nucleoside and nucleotide compounds in the treatment of these conditions, see, for example, Neuropsychopharmacology. 2015 Jan;40( 2):305-14. doi: 10.1038/npp.2014.173. Epub 2014 Jul 15. Impaired cognition after stimulation of a P2Yi receptor in the rat medial prefrontal cortex, Koch, H. et al. PMID: 25027332, which is included herein description by reference). In other embodiments, the disease or condition is selected from a neurodegenerative disease such as Alzheimer's disease, Parkinson's disease, Huntington's disease, prion disease, and/or amyotrophic lateral sclerosis (for information on the use of certain nucleoside and nucleotide compounds in the treatment of these conditions, see, e.g., US 8691775, which is incorporated herein by reference). In other embodiments, the disease or condition is selected from hearing impairment, Meniere's disease, endolymphatic hydrops, progressive hearing loss, dizziness, vertigo, tinnitus, concomitant brain injury associated with radiation therapy for cancer, and/or treatment of migraine (information on the use of certain nucleoside and nucleotide compounds in the treatment of these conditions, see, for example, US 2009/0306225; UY31779 and US 8399018, each of which is incorporated herein by reference). In other embodiments, the disease or condition is selected from sleep disorders, depression, sleep disorders in the elderly, Parkinson's disease, Alzheimer's disease, epilepsy, schizophrenia, and/or symptoms occurring in recovering alcoholics (information on the use of certain nucleoside and nucleotide compounds in the treatment for these conditions, see, for example, US 2014/0241990, which is incorporated herein by reference). In other embodiments, the disease or condition is selected from injury to neurons or nerves of the peripheral nervous system during surgery (for information on the use of certain nucleoside and nucleotide compounds in the treatment of these conditions, see, for example, US 8,685,372, which is incorporated herein by reference). . In other embodiments, the disease or condition is cancer, such as prostate cancer (for information on the use of certain nucleoside and nucleotide compounds in the treatment of these conditions, see, for example, Biochem Pharmacol. 2011 August 15; 82(4): 418- 425. doi:10.1016/j.bcp.2011.05.013, Activation of the P2Y1 Receptor Induces Apoptosis and Inhibits Proliferation of Prostate Cancer Cells, Qiang Wei et al., which is incorporated herein by reference). In other embodiments, the disease or condition is selected from one or more gastrointestinal disorders, such as constipation and/or diarrhea (for information on the use of certain nucleoside and nucleotide compounds in the treatment of these conditions, see, for example, Acta Physiol (Oxf) 2014 Dec;212(4):293-305. doi: 10.1111/apha.l2408. Differential functional role of purinergic and nitrergic inhibitory cotransmitters in human colonic relaxation, Mane N1, Gil V, Martinez-Cutillas M, Clave P, Gallego D, Jimenez M.; and Neurogastroenterol. Motil. 2014 Jan;26(1): 115-23. doi: 10.1111/nmo.12240. Epub 2013 Oct 8. Calcium responses in subserosal interstitial cells of the guinea-pig proximal colon, Tamada H., Hashitani H. PMID: 24329947, which is incorporated herein by reference). In other embodiments, the disease or condition is selected from CNS-mediated pain, such as neuropathic pain, inflammatory pain, and/or acute pain (for information on the use of certain nucleoside and nucleotide compounds in the treatment of these conditions, see, for example, Br J Pharmacol. 2010 Mar;159(5): 1106-17. doi: 10.1111/j.1476-5381.2009.00596.x. Epub 2010 Feb 5. A comparative analysis of the activity of ligands acting at P2X and P2Y receptor subtypes in models of neuropathic , acute and inflammatory pain And0 RD1, Mehesz B, Gyires K, Illes P, Sperlagh B. PMID: 20136836, which is incorporated herein by reference).

In other embodiments, the disease or condition is selected from brain cancer, such as glioblastoma (for information on the use of certain nucleoside and nucleotide compounds in the treatment of these conditions, see, for example, Purinergic Signal. 2015 Sep;11(3):331-46 doi:

- 18 046380

10.1007/s11302-015-9454-7. Epub 2015 May 15. Potentiation of temozolomide antitumor effect by purine receptor ligands able to restrain the in vitro growth of human glioblastoma stem cells. D'Alimonte, I. et al. PMID: 25976165, which is incorporated herein by reference). In other embodiments, the disease or condition is pain (for information on the use of certain nucleoside and nucleotide compounds in the treatment of pain, see, for example, Pharmacol Biochem Behav. 2015 Jan;128:23-32. doi: 10.1016/j.pbb. 2014.11.001. Epub 2014 Nov 6. Participation of peripheral P2Y1, P2Y ₆ and P2Yn receptors in formalin-induced inflammatory pain in rats. Barragan-Iglesias P. et al. PMID: 25449358; and Neuropharmacology. 2014 Apr;79:368- 79. doi: 10.1016/j.neuropharm.2013.12.005. Epub 2013 Dec 12. Blockade of peripheral P2Yi receptors prevents the induction of thermal hyperalgesia via modulation of TRPV1 expression in carrageenan-induced inflammatory pain rats: involvement of p38 MAPK phosphorylation in DRGs Kwon SG, Roh DH, Yoon SY, Moon JY, Choi SR, Choi HS, Kang SY, Han HJ, Beitz AJ, Lee JH (PMID: 24333674, which is incorporated herein by reference). In other embodiments, the disease or condition is selected from a gastrointestinal disorder such as diarrhea (for information on the use of certain nucleoside and nucleotide compounds in the treatment of these conditions, see, for example, Acta Physiol (Oxf). 2014 Dec;212(4 ):293-305. doi: 10.1111/apha. 12408. Differential functional role of purinergic and nitrergic inhibitory cotransmitters in human colonic relaxation, Mane N., Gil V, Martinez-Cutillas M., Clave P., Gallego D., Jimenez M., which is incorporated herein by reference). In other embodiments, the disease or condition is a cognitive impairment (for information on the use of certain nucleoside and nucleotide compounds in the treatment of this condition, see, for example, Neuropsychopharmacology. 2015 Jan;40(2):305-14. doi: 10.1038/ npp.2014.173. Epub 2014 Jul 15. Impaired cognition after stimulation of P2Y1 receptors in the rat medial prefrontal cortex, Koch H, Bespalov A, Drescher K, Franke H, Krugel U. PMID: 25027332, which is incorporated herein by reference) .

In some embodiments, the present invention provides a method of treating a disease or condition associated with brain damage or a neurodegenerative condition, such as epilepsy, migraine, concomitant brain damage associated with radiation therapy for cancer, depression, mood or behavioral changes, dementia, erratic behavior , suicidal tendencies, tremors, Huntington's chorea, decreased motor coordination, hearing loss, speech disorders, dry eye syndrome, hypomia, attention deficit, memory loss, cognitive impairment, vertigo, dysarthria, dysphagia, visual disturbances or disorientation, including administration to someone in need to the patient an effective amount of the disclosed compound. In some embodiments, the compound is an A ₃ R agonist. In some embodiments, the compound is a P2Y1 agonist. In some embodiments, the compound is a potency-biased A3 receptor agonist, an A3 partial agonist, or a potency-biased partial A3 agonist, or an A3 antagonist. In some embodiments, the compound acts through dual agonism at A ₃ R and A1R. In some embodiments, the compound is a potency-biased P2Y1 receptor agonist, a partial P2Y1 agonist, or a potency-biased partial P2Y1 agonist, or a P2Y1 antagonist. In some embodiments, the compound is a solid form of Compound A or a pharmaceutically acceptable composition thereof.

In additional embodiments, the present invention provides a method of treating a neurodegenerative disease selected from the group consisting of Alzheimer's disease, Parkinson's disease, Huntington's disease, multiple sclerosis, amyotrophic lateral sclerosis, and prion disease in a patient in need thereof, comprising administering an effective amount of a disclosed compound. In some embodiments, the compound is an A3R agonist. In some embodiments, the compound is a P2Y 1 agonist. In some embodiments, the compound is a potency-biased A3 receptor agonist, an A3 partial agonist, or a potency-biased partial A3 agonist, or an A3 antagonist. In some embodiments, the compound acts through dual agonism at A ₃ R and A1R. In some embodiments, the compound is a potency-biased P2Y1 receptor agonist, a partial P2Y1 agonist, or a potency-biased P2Y1 partial agonist, or a P2Y 1 antagonist. In some embodiments, the compound is a solid form of Compound A or a pharmaceutically acceptable composition thereof.

In some embodiments, an improvement in cognitive or neurological function is defined as an increase in score from about 1% to 20% on the delayed verbal recall task of the Wechsler Memory Scale-Revised. For example, an improvement in cognitive function may be defined as an increase in score from about 1% to 10% or from about 1% to 5%.

In some embodiments, the present invention provides a method of treating a brain or central nervous system (CNS) injury or condition selected from traumatic brain injury (TBI) or stroke, comprising administering to a patient in need thereof an effective amount of a solid form of Compound A or a pharmaceutically acceptable form containing the same

- 19 046380 compositions.

In some embodiments, the injury or condition of the brain or central nervous system (CNS) is a TBI. In some embodiments, the TBI is selected from concussion, blast injury, combat trauma, or mild, moderate, or severe blow to the head.

In some embodiments, a solid form of Compound A or a pharmaceutically acceptable composition containing it is administered within 24 hours of TBI or stroke.

In some embodiments, a solid form of Compound A or a pharmaceutically acceptable composition containing it is administered within 8 hours of TBI or stroke.

In some embodiments, a solid form of Compound A or a pharmaceutically acceptable composition containing it is administered for at least the first 8 to 48 hours after TBI or stroke.

In some embodiments, the injury or condition of the brain or central nervous system (CNS) is a stroke.

In some embodiments, a solid form of Compound A or a pharmaceutically acceptable composition containing it is chronically administered to treat stroke over a period of time from the onset of the stroke until resolution of the stroke.

In some embodiments, the patient experiences increased neuroprotection or recovery of neural tissue function compared to that of a patient not receiving treatment.

In some embodiments, the solid form of Compound A, or a pharmaceutically acceptable composition containing it, is a partial agonist of the human adenosine A3 receptor (A3R) with biased activity. In some embodiments, the compound acts through dual agonism at A3R and A1R.

In some embodiments, the A3R is subject to partial agonism biased toward the neuroprotective functions of the A3R receptor.

In some embodiments, the solid form of Compound A, or a pharmaceutically acceptable composition containing it, is administered orally, intravenously, or parenterally.

In one aspect, the present invention provides a method of enhancing neuroprotection or restoring neural tissue function in a patient who has suffered a TBI or stroke, comprising administering to the patient in need thereof an effective amount of a solid form of Compound A or a pharmaceutically acceptable composition containing the same.

In some embodiments, neuroprotection or restoration of neural tissue function provides a shorter recovery period after TBI or stroke compared to that of an untreated patient. In some embodiments, the solid form of Compound A, or a pharmaceutically acceptable composition containing it, is a partial agonist of the human adenosine A3 receptor (A3R) with biased activity, and the A3R is subject to partial agonism biased toward the neuroprotective functions of the A3R receptor. In some embodiments, the compound acts through dual agonism at A ₃ R and A _| R

In some embodiments, the solid form of Compound A, or a pharmaceutically acceptable composition containing it, is administered orally, intravenously, or parenterally.

In one aspect of the present invention, there is provided a method of treating an injury, disease or condition selected from traumatic brain injury (TBI), stroke, neurodegenerative condition, or heart or cardiovascular disease, comprising administering to a patient in need thereof an effective amount of a solid form of Compound A or containing a pharmaceutically acceptable composition thereof.

In some embodiments, the injury, disease, or condition is a TBI. In some embodiments, the TBI is selected from concussion, blast injury, combat trauma, or mild, moderate, or severe blow to the head.

In some embodiments, the injury, disease, or condition is a stroke selected from ischemic stroke, hemorrhagic stroke, subarachnoid hemorrhage, cerebral vasospasm, or transient ischemic attack (TIA).

In some embodiments, the neurodegenerative disease is selected from Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), multiple sclerosis (MS), amyotrophic lateral sclerosis (ALS), chronic traumatic encephalopathy (CTE), or a neurodegenerative condition caused by a virus, alcoholism, tumor, toxin, or repeated brain damage.

In some embodiments, the injury, disease, or condition is Parkinson's Disease.

In some embodiments, the injury, disease, or condition is Alzheimer's disease, migraine, brain surgery, or a neurological side effect due to cancer chemotherapy.

In some embodiments, the heart disease or cardiovascular disease you

- 20 046380 for cardiac ischemia, myocardial infarction, cardiomyopathy, coronary heart disease, arrhythmia, myocarditis, pericarditis, angina pectoris, hypertensive heart disease, endocarditis, rheumatic heart disease, congenital heart disease or atherosclerosis.

In some embodiments, the heart disease or cardiovascular disease is cardiac ischemia or myocardial infarction.

In some embodiments, a solid form of Compound A or a pharmaceutically acceptable composition containing it is chronically administered to treat stroke, cardiac ischemia, or myocardial infarction over a period of time from the onset of the injury to its resolution.

In some embodiments, the patient experiences increased neuroprotection or recovery of neural tissue function compared to that of a patient not receiving treatment.

In some embodiments, the A3R is subject to partial agonism biased toward the neuroprotective functions of the A3R receptor through preferential activation of intracellular calcium mobilization with less or no activation of other A3R-mediated signaling pathways, or through preferential activation of GqH-mediated intracellular calcium mobilization, Gi-mediated modulation cAMP production or Gi-mediated phosphorylation of ERK1/2 and Akt.

In some embodiments, A ₃ R is subject to partial agonism biased toward the cardioprotective functions of the A ₃ R receptor, through preferential activation of intracellular calcium mobilization with less or no activation of other A ₃ R-mediated signaling pathways, or through preferential activation of GqH-mediated intracellular calcium mobilization. calcium, Gi-mediated modulation of cAMP production, or Gi-mediated phosphorylation of ERK1/2 and Akt.

In some embodiments, the method provides increased neuroprotection or restoration of neural tissue function in a patient suffering from a neurological side effect caused by or resulting from cancer chemotherapy or brain surgery. In some embodiments, the solid form of Compound A or a pharmaceutically acceptable composition containing it is administered orally. In one aspect of the present invention, there is provided a method of enhancing neuroprotection or restoring neural tissue function in a patient who has suffered a TBI or stroke, thereby providing a method of treating TBI or stroke, comprising administering to a patient in need thereof an effective amount of a solid form of Compound A or a pharmaceutically acceptable composition containing the same. .

In one aspect of the present invention, there is provided a method of enhancing cardioprotection or regeneration of damaged heart tissue in a patient who has suffered cardiac ischemia or myocardial infarction, thereby providing a method of treating cardiac ischemia or myocardial infarction, comprising administering to the patient in need thereof an effective amount of a solid form of Compound A or containing a pharmaceutically acceptable composition thereof.

In some embodiments, the recovery period from TBI, stroke, cardiac ischemia, or myocardial infarction is reduced compared to that of an untreated patient.

In some embodiments, the A ₃ R is subject to partial agonism biased toward the neuroprotective functions of the A ₃ R receptor.

In some embodiments, the A ₃ R is subject to partial agonism biased toward the cardioprotective functions of the A ₃ R receptor.

In some embodiments, the solid form of Compound A or a pharmaceutically acceptable composition containing it is administered orally.

In some embodiments, the compound is a potency-biased A3R agonist with improved cardioprotective function compared to a full A3R agonist.

In some embodiments, the solid form of Compound A, or a pharmaceutically acceptable composition containing it, is an A3R agonist with biased activity with improved cardioprotective function compared to a full _A3R agonist, mediated by preferential activation of one or more of the following A3R-mediated signaling pathways: activation GqH-mediated mobilization intracellular calcium, Gi-mediated modulation of cAMP production, Gi-mediated phosphorylation of ERK1/2 and Akt, or modulation of beta-arrestin activation.

In some embodiments, the solid form of Compound A or a pharmaceutically acceptable composition containing it is an A ₃ R agonist with biased activity with improved cardioprotective function compared to a full A3R agonist, mediated by preferential activation of intracellular calcium mobilization with less or no activation of other A3R-mediated signaling pathways such.

In some embodiments, the solid form of Compound A or a pharmaceutically acceptable composition containing it is a partial A3R agonist with improved cardioprotective function compared to a full A3R agonist.

Addictive disorders

The disclosed compounds are also suitable for use in the treatment of addictions, addictive

- 21 046380 behavior, behavioral addictions, compulsive disorders and compulsive behavior and related conditions.

The use of compounds such as Compound A in the treatment of such addictions, behaviors and disorders is described in WO/2019/157317, the contents of which are incorporated herein by reference. Mice self-administering cocaine exhibit significantly higher levels of glutamate in the VTA (ventral tegmental area) of the brain. VTAs, particularly VTA dopamine neurons, have several functions in reward, motivation, cognition, and drug addiction and may be a focus of attention in some psychiatric disorders. The increased glutamate content appears to be due, at least in part, to decreased glutamate uptake in astrocytes. Without being bound by theory, it is believed that decreased glutamate availability negatively affects astrocyte function, and this decreased function influences neuronal activity and drug-seeking behavior. The compounds disclosed herein have now been found to provide treatment or prevention of relapse in individuals suffering from addiction, for example, by reversing such decline in astrocyte function. This reduction in astrocyte function may be due in part to decreased expression of the glutamate transporter (GLT-1) in astrocytes. Because astrocytes metabolize glutamate to produce ATP, this likely impairs glutamate uptake, impairs astrocyte oxidative metabolism and subsequent ATP-dependent processes, and thereby impairs the ability of astrocytes to maintain an environment optimal for VTA neuronal activity.

Accordingly, in one aspect of the present invention there is provided a method of preventing, ameliorating, treating or accelerating recovery from addiction, addictive behavior, behavioral dependence, brain reward system disorder, compulsive disorder or a related condition, comprising administering to a subject in need thereof an effective amount of a solid form of the compound A or a pharmaceutically acceptable composition containing it.

In some embodiments, the addiction is a dependence on an addictive substance. In some embodiments, the addictive substance is a prescription drug or a recreational drug.

In some embodiments, the addictive substance is selected from alcohol, nicotine, a stimulant, a cannabinoid receptor agonist, or an opioid receptor agonist. In some embodiments, the addictive substance is selected from heroin, cocaine, alcohol, a volatile drug, an opioid, nicotine, amphetamine or a synthetic analogue, a salt, a composition, or a combination thereof.

In some embodiments, the amphetamine is selected from bupropion, cathinone, methylenedioxymethamphetamine (MDMA), or methamphetamine.

In some embodiments, the prescription drug or recreational drug is selected from a cannabinoid receptor agonist or an opioid receptor agonist.

In some embodiments, the addiction is alcohol or nicotine addiction.

In some embodiments, the subject suffers from a combined substance abuse disorder.

In some embodiments, the prescription drug or recreational drug is selected from cocaine, heroin, bupropion, cathinone, MDMA or methamphetamine, morphine, oxycodone, hydromorphone, fentanyl, or a combination thereof.

In some embodiments, the disclosed compound enhances astrocyte-mediated energy metabolism, such as astrocyte mitochondria. In some embodiments, the compound reverses the reduction in glutamate uptake in astrocytes caused by a substance with addictogenic potential.

In some embodiments, the compound at least partially reverses the remodeling of the brain's reward system. In some embodiments, such effects are mediated by brain or CNS A ₃ adenosine receptors, such as the A ₃ R of astrocytes in the VTA or the A 3 R of microglia.

In another aspect of the present invention, there is provided a method of preventing, ameliorating, treating, or accelerating recovery from addiction, addictive behavior, behavioral addiction, brain reward system disorder, compulsive disorder, or related condition by enhancing energy metabolism mediated by astrocytes, glia, microglia, neurons, endothelial cells or other cells of the brain and/or central nervous system, comprising administering to a subject in need thereof an effective amount of a solid form of Compound A or a pharmaceutically acceptable composition containing the same. In some embodiments, the method provides treatment or prevention of relapse of addiction or addictive behavior in a subject. In some embodiments, the subject is addicted to one or more addictive substances, such as addictive drugs (drugs having addictive potential). As described below, such substances include formulations

- 22 046380 medicinal and recreational drugs such as heroin, cocaine, nicotine or opioid agonist. In another aspect, the present invention provides a method of treating or preventing withdrawal symptoms caused by dependence on one or more addictive substances or drugs, comprising administering to a subject in need thereof an effective amount of a solid form of Compound A or a pharmaceutically acceptable composition containing the same. In some embodiments, the compound provides a reduction in the severity of withdrawal symptoms in a person suffering from addiction and in a state of withdrawal. In some embodiments, the compound provides treatment for withdrawal symptoms in a person suffering from addiction and in a state of withdrawal. In some embodiments, the method further includes co-administration of another drug to treat withdrawal symptoms and, optionally, counseling such as psychotherapy. In some embodiments, the method further includes cognitive behavioral therapy. In some embodiments, the method further includes a digital therapeutic agent. Digital therapeutics include, for example, reSET or reSET-O (Pear Therapeutics).

In some embodiments, the present invention provides a method of treating or preventing the relapse of a compulsive disorder or compulsive behavior, comprising administering to a subject in need thereof an effective amount of a solid form of Compound A or a pharmaceutically acceptable composition containing the same.

In some embodiments, the compulsive disorder is obsessive-compulsive disorder (OCD), Tourette's syndrome, trichotillomania, anorexia, bulimia, anxiety disorder, psychosis, or post-traumatic stress disorder.

In accordance with another aspect of the present invention, there is provided a method of treating one or more types of behavioral addiction and addictive behavior or addictive disorder, comprising administering to a subject in need thereof a solid form of Compound A or a pharmaceutically acceptable composition containing it. Behavioral addictions and addictive disorders result from the intoxication that the subject feels as a result of the release of chemicals in the brain (for example, serotonin, adrenaline, epinephrine, and the like) during certain activities. Such disorders are known in the art and include gambling addiction, sex addiction, pornography addiction, eating disorders, spending addictions, rage/anger, workaholism, exercise addiction, risky behavior addictions (e.g., kleptomania and pyromania). ), perfectionism, internet or video game addiction, and uncontrolled use of electronic devices such as texting and checking social media, to name a few.

In some embodiments, activation of astrocytes is achieved by bringing one or more purinergic receptors, such as adenosine receptors (AR), such as those associated with or expressed by astrocytes or microglia, into contact with an exposed compound, thereby modulating the activity of the one or more receptors. In some embodiments, the compound activates astrocytes by acting on adenosine receptors on astrocytes, such as A1, A2A, A2B, and A3 receptors, to treat one or more disclosed diseases or conditions. In some embodiments, upon administration to a subject in need thereof, the disclosed compound affects one or more functions, such as glutamate uptake, affecting astrocyte energy metabolism or neuronal function, thereby providing treatment for one or more diseases or conditions. In some embodiments, the compound is an AR agonist. In some embodiments, the purinergic receptor is an adenosine A ₃ receptor (A ₃ R). In some embodiments, the compound is an _A3R agonist. In some embodiments, the compound is an A3 receptor (A3R) partial agonist, such as the human A3 receptor (hA3R), or a potency-biased A3R agonist, or a potency-biased _A3R partial agonist. activity. In some embodiments, the compound is an A ₃ receptor antagonist with biased activity. In some embodiments, the compound acts through dual agonism at A3R and A1R. In some embodiments, the compound is a solid form of Compound A or a pharmaceutically acceptable composition thereof.

P2Y receptors are G protein-coupled receptors, and various subtypes of these receptors play important roles in processes such as synaptic communication, cell differentiation, ion flow, vasodilation, blood-brain barrier permeability, platelet aggregation, and neuromodulation. Characteristic members of the P2Y purinergic receptor family include the mammalian P2Y ₁ , P2Y ₁₁ , P2Y ₁₂ and P2Y13 receptors, which bind to adenine nucleotides; receptors P2Y4, P2Y6 and P2Y ₁₄ , which bind to uracil nucleotides; and rodent P2Y2 receptors and P2Y4 receptors, which have mixed selectivity. In some embodiments, activation of astrocytes is achieved by driving one or more purinergic receptors, such as P2Y receptors, such as those associated with astrocytes

- 23 046380 or expressed by them, into contact with the disclosed compound, which provides modulation of the activity of one or more receptors. In some embodiments, the compound activates astrocytes by targeting P2Y receptors, such as P2Y1, P2Y11, P2Y12, and P2Y13 receptors associated with or expressed by astrocytes, to treat one or more disclosed diseases or conditions. In some embodiments, the P2Y receptor is a P2Y receptor _| . In some embodiments, the P2Y1 receptor is located on intracellular mitochondrial membranes. In some embodiments, the compound is a P2Y agonist. In some embodiments, the compound is a P2Y agonist _| eg P2Y receptor agonist _| person. In some embodiments, the compound is a P2Y| receptor agonist, such as an activity-biased human P2Y1 receptor, a P2Y1 partial agonist, or a P2Y| partial agonist. with shifted activity. In some embodiments, the compound is a P2Y|receptor antagonist. with shifted activity. In some embodiments, the compound is a solid form of Compound A or a pharmaceutically acceptable composition thereof.

As used herein, the term addiction includes, unless otherwise specified, physical or psychological dependence on a substance. Addiction may include withdrawal symptoms or mental or physical suffering if the substance is withdrawn. Addiction includes drug addiction, drug dependence, habituation, neurological and/or synaptic changes, development of disturbances in the brain's reward system, behavioral changes, or other signs or symptoms of addiction in the subject.

As used herein, the term addictive drug or drug having addictogenic potential includes drugs and other substances, such as nicotine, whether approved by a regulatory authority for the treatment of a disease or not, which are known to cause to clinical, behavioral or neurological manifestations of addiction or compulsive behavior. In some embodiments, the addictive drug includes nicotine, a cannabinoid receptor agonist, a stimulant, or an opioid receptor agonist. Addictive substance refers to addictive drugs as well as other substances of abuse, such as alcohol. Examples of addictive substances include heroin, cocaine, alcohol, opiates, nicotine, volatile drugs, amphetamines and their synthetic analogues.

Pharmaceutically acceptable compositions

According to another embodiment of the present invention, there is provided a composition containing the disclosed compound and a pharmaceutically acceptable carrier, adjuvant or transport medium. In some embodiments, a composition of the present invention is formulated for administration to a patient in need of such a composition. In some embodiments, the composition of the present invention is formulated for oral administration to a patient.

The term biological sample as used herein includes, without limitation, cell cultures or extracts thereof; biopsy material obtained from a mammal, or extracts thereof; and blood, saliva, urine, feces, semen, tears or other body fluids or extracts thereof.

The term patient as used herein means an animal, preferably a mammal, and most preferably a human.

The term pharmaceutically acceptable carrier, adjuvant or vehicle refers to a non-toxic carrier, adjuvant or vehicle that does not interfere with the pharmacological activity of the compound with which it is formulated. Pharmaceutically acceptable carriers, adjuvants or transfer media that may be used in the compositions of the present invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffers such as phosphates, glycine, sorbic acid, potassium sorbate, mixtures of partial glycerides of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium phosphate, dipotassium phosphate, sodium chloride, zinc salts, colloidal silicon dioxide, magnesium trisilicate, polyvinylpyrrolidone , cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyoxyethylene-polyoxypropylene block copolymers, polyethylene glycol and lanolin.

Pharmaceutically acceptable derivative means any non-toxic salt, ester, ester salt or other derivative of a compound of the present invention which, upon administration to a recipient, is capable of directly or indirectly providing the presence of a compound of the present invention or a metabolite or residue thereof having inhibitory activity.

The compounds and compositions, in accordance with the method according to the present invention, are administered using any amount and any route of administration effective for treating or reducing

- 24 046380 severity of the disorder mentioned above. The exact amount required will vary from subject to subject, depending on the species, age and general condition of the subject, severity of the infection, the specific agent, route of administration, and the like. The compounds of the present invention are preferably formulated in unit dosage form for ease of administration and uniformity of dosage. The expression unit dosage form as used herein refers to a physically discrete unit of agent suitable for the patient being treated. It should be understood, however, that the general daily use of the compounds and compositions of the present invention will be determined by the attending physician within the bounds of sound medical judgment. The specific effective dose level for any particular patient or organism will depend on a variety of factors, including the disorder being treated and the severity of the disorder; the activity of the particular compound used; the specific composition used; the patient's age, weight, general health, gender, and diet; time, route and speed of withdrawal of the specific connection used; duration of treatment; drugs used in combination or coincident with the particular compound used, and similar factors well known in medicine.

The pharmaceutically acceptable compositions of the present invention can be administered to humans and other animals orally, rectally, parenterally, intracisternally, intravaginally, intraperitoneally, topically (eg, as powders, ointments, or drops), buccally, as oral or nasal sprays, or the like. , depending on the severity of the infection being treated. In some embodiments, the compounds of the present invention are administered orally or parenterally at a dose level of from about 0.01 mg/kg to about 50 mg/kg, and preferably from about 0.01 mg/kg to about 25 mg/kg of the subject's body weight in day, one or more pA3 per day, to obtain the desired therapeutic effect. In some embodiments, the compounds of the present invention are administered orally or parenterally at a dose level of from about 0.01 mg/kg to about 50 mg/kg, or from about 0.01 mg/kg to about 25 mg/kg, or from about 0 .05 mg/kg to about 10 mg/kg, or about 0.05 mg/kg to about 5 mg/kg, or about 0.1 mg/kg to about 2.5 mg/kg of the subject's body weight per day , one or more pA3 per day, to obtain the desired therapeutic effect.

Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, liposomes, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate , propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (especially cottonseed, peanut, corn, wheat germ, olive, castor and sesame oils), glycerin, tetrahydrofurfuryl alcohol, polyethylene glycol and sorbitan fatty acid esters, and their mixtures. In addition to inert diluents, oral compositions may also include substances such as wetting agents, emulsifying and suspending agents, sweetening, flavoring and perfuming agents.

Injectable preparations, for example sterile injectable aqueous or oleaginous suspensions, can be prepared in accordance with what is known in the art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in a non-toxic parenterally acceptable diluent or solvent, for example, a solution in 1,3-butanediol. Acceptable carriers and solvents that may be used include water, USP Ringer's solution, and isotonic sodium chloride solution. In addition, sterile non-volatile oils are usually used as a solvent or suspending medium. Any light fatty oil, including synthetic mono- or diglycerides, can be used for this purpose. In addition, fatty acids such as oleic acid are used to obtain injection preparations.

Injectable formulations can be sterilized, for example, by filtration through a bacteria-retaining filter or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved or dispersed in sterile water or other sterile injectable media prior to use.

To prolong the effect of the compound of the present invention, it is often desirable to slow down the absorption of the compound from the site of subcutaneous or intramuscular administration. This can be achieved by using a liquid suspension of a crystalline or amorphous mass with poor water solubility. The rate of absorption of a compound accordingly depends on its rate of dissolution, which in turn may depend on crystal size and crystalline form. Alternatively, delayed absorption of the parenterally administered compound form is achieved by dissolving or suspending the compounds in an oil vehicle. Injection depot forms are made by forming microencapsular matrices of compounds in biodegradable poly

- 25 046380 measures such as polylactide-polyglycolide. Depending on the ratio of the compound to the polymer and the nature of the particular polymer used, the rate of release of the compound can be adjusted. Examples of other biodegradable polymers include poly(orthoesters), poly(anhydrides), as well as cyclodextrins and modified cyclodextrins (such as SBE-bCD). Injectable depot formulations are also prepared by incorporating the compound into liposomes or microemulsions that are compatible with body tissues.

Compositions for rectal or vaginal administration are preferably suppositories, which can be prepared by mixing the compounds of the present invention with suitable non-irritating excipients or carriers, such as cocoa butter, polyethylene glycol or suppository wax, which are solid at ambient temperature, but liquid at body temperature and thus melt in the rectum or vaginal cavity and release the active compound. Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In such solid dosage forms, the active compound is mixed with at least one inert pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or a) excipients or diluents such as starches, lactose, sucrose, glucose, mannitol and silicic acid , b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose and gum arabic) humectants such as glycerin, d) various agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid , some silicates and sodium carbonate, e) dissolution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium, g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also contain buffering agents.

Solid compositions of this type can also be used as fillers in soft and hard gelatin capsules using excipients such as lactose or milk sugar, as well as high molecular weight polyethylene glycols and the like. Solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared with coatings and coatings, such as enteric coatings and other coatings well known in the pharmaceutical art. They may optionally contain opacifying agents, and may also be of a composition in which they release the active ingredients only or preferentially in a certain part of the intestinal tract, optionally with a delay. Examples of excipient compositions that can be used include polymeric substances and waxes. Solid compositions of this type can also be used as fillers in soft and hard gelatin capsules using excipients such as lactose or milk sugar, as well as high molecular weight polyethylene glycols and the like.

The active compounds may also be in microencapsulated form with one or more excipients, as noted above. Solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared with coatings and coatings, such as enteric coatings, controlled release coatings and other coatings well known in the pharmaceutical formulation art. In such solid dosage forms, the active compound may be admixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms may also contain, as is common practice, foreign substances other than inert diluents, for example, tableting lubricants and other tabletting aids such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, dosage forms may also contain buffering agents. They may optionally contain opacifying agents, and may also be of a composition in which they release the active ingredients only or preferentially in a certain part of the intestinal tract, optionally with a delay. Examples of excipient compositions that can be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of the compounds of this invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalers or patches. The active ingredient is admixed under sterile conditions with a pharmaceutically acceptable carrier and any necessary preservatives or buffers that may be required. Ophthalmic composition, ear drops and eye drops are also considered to be within the scope of this invention. In addition, the present invention involves the use of transdermal patches, which have the additional advantage of providing controlled delivery of compounds into the body. Such dosage forms can be made by dissolving or dosing the compound in the correct medium. Absorption enhancers can also be used to increase the flux of compounds through the skin. Rate control can be accomplished either by providing a rate-controlling membrane or by dispersing the compound in a polymer matrix

- 26 046380 or gel.

The compounds of the present invention can also be administered topically, eg directly into the eye, eg in the form of eye drops or ophthalmic ointment. Eye drops typically contain an effective amount of at least one compound of the present invention and a carrier that is capable of being safe when applied to the eye. For example, eye drops are in the form of an isotonic solution, and the pH of the solution is adjusted so that there is no irritation to the eyes. In many cases, the epithelial barrier prevents molecules from entering the eye. Thus, most currently used ophthalmic drugs contain some form of penetration enhancer. These penetration enhancers act by weakening the tight junctions of the epithelial cells closest to the surface (Burstein, 1985, Trans Ophthalmol Soc U K 104(Pt 4): 402-9; Ashton et al., 1991, J Pharmacol ExpTher 259(2): 719- 24; Green et al., 1971, Am J Ophthalmol 72(5): 897-905). The most commonly used penetration enhancer is benzalkonium chloride (Tang et al., 1994, J Pharm Sci 83(1): 85-90; Burstein et al, 1980, Invest Ophthalmol Vis Sci 19(3): 308-13), which also acts as a preservative to prevent contamination by microorganisms. It is usually added to a final concentration of 0.01-0.05%.

Combinations with other therapeutic agents

Depending on the particular condition or disease being treated, additional therapeutic agents that are typically administered to treat that condition may also be present in the compositions of the present invention. As used herein, additional therapeutic agents that are typically administered to treat a particular disease or condition are known to be relevant to the disease or condition being treated. In some embodiments, the present compound or composition thereof is administered to a patient in need thereof in combination with other therapeutic agents, such as tissue plasminogen activators, anticoagulants, statins, ACE inhibitors, angiotensin II receptor blockers (ARBs), beta blockers, calcium channel blockers or diuretics. In some embodiments, tissue plasminogen activators used in combination with the compounds or compositions of the present invention include, but are not limited to, alteplase, desmoteplase, reteplase, tenecteplase, or combinations of any of the above.

In some embodiments, anticoagulants used in combination with the compounds or compositions of the present invention include, but are not limited to, warfarin, heparin, apixaban, clopidogrel, aspirin, rivaroxaban, dabigatran, or combinations of any of the above. In some embodiments, statins used in combination with the compounds or compositions of the present invention include, but are not limited to, atorvastatin, rosuvastatin, fluvastatin, lovastatin, pravastatin, simvastatin and pitavastatin, cerivastatin, mevastatin, or combinations of any of the above.

In some embodiments, ACE inhibitors used in combination with the compounds or compositions of the present invention include, but are not limited to, captopril, enalapril, fosinopril, lisinopril, moexipril, perindopril, quinapril, ramipril, trandolapril, benazepril, or combinations of any of the above .

In some embodiments, angiotensin II receptor blockers (ARBs) used in combination with the compounds or compositions of the present invention include, but are not limited to, azilsartan, candesartan, eprosartan, irbesartan, losartan, olmesartan, telmisartan, valsartan, fimasartan, or combinations of any from those mentioned above.

In some embodiments, beta blockers used in combination with the compounds or compositions of the present invention include, but are not limited to, atenolol, bisoprolol, betaxolol, carteolol, carvedilol, labetalol, metoprolol, nadolol, nebivolol, oxprenolol, penbutolol, pindolol, propranolol, timolol or combinations of any of the above.

In some embodiments, calcium channel blockers used in combination with the compounds or compositions of the present invention include, but are not limited to, dihydropyridines: amlodipine, cilnidipine, clevidipine, felodipine, isradipine, lercanidipine, levamlodipine, nicardipine, nifedipine, nimodipine, nisoldipine, nitrendipine, diltiazem, verapamil or combinations of any of the above.

In some embodiments, diuretics used in combination with the compounds or compositions of the present invention include, but are not limited to, loop diuretics, thiazide diuretics, thiazide-like diuretics and potassium-sparing diuretics, or combinations of any of the above.

In some embodiments, loop diuretics used in combination with the compounds or compositions of the present invention include, but are not limited to, bumetanide, ethacrynic acid, furosemide, torsemide, or combinations of any of the above.

In some embodiments, thiazide diuretics used in combination with the compounds or compositions of the present invention include, but are not limited to, epitizide, hydrochlorothiazide and chlorothiazide, bendroflumethiazide, methyclothiazide, polythiazide, or a combination

- 27 046380 nations of any of the above.

In some embodiments, thiazide-like diuretics used in combination with the compounds or compositions of the present invention include, but are not limited to, indapamide, chlorthalidone, metolazone, or combinations of any of the above.

In some embodiments, potassium-sparing diuretics used in combination with the compounds or compositions of the present invention include, but are not limited to, amiloride, triamterene, spironolactone, eplerenone, or combinations of any of the above.

In some embodiments, the present compound or composition thereof is administered to a patient in need thereof in combination with a mechanical thrombectomy device. In some embodiments, the mechanical thrombectomy device is a stroke thrombectomy device or a coil embolization device for cerebral aneurysm. In some embodiments, such a device includes, but is not limited to, an aspiration device or a stent retriever.

In some embodiments, a combination of 2 or more therapeutic agents may be co-administered with the compounds or compositions of the present invention. In some embodiments, a combination of 3 or more therapeutic agents may be co-administered with the compounds or compositions of the present invention.

These additional agents may be administered separately from a composition containing a compound of the present invention as part of a multiple dosing regimen. Alternatively, these agents may be part of a single dosage form, being mixed together with a compound of the present invention to form a single composition. When administered as part of a multiple-dose regimen, the two active agents may be delivered simultaneously, sequentially, or spaced apart, typically within five hours from administration of one to administration of the other. As used herein, the term combination, combined and related terms refer to the simultaneous or sequential administration of therapeutic agents according to the present invention. For example, a compound of the present invention may be co-administered with another therapeutic agent simultaneously or sequentially in separate unit dosage forms or together in a single unit dosage form. Accordingly, the present invention provides a single unit dosage form containing a compound of the present invention, an additional therapeutic agent, and a pharmaceutically acceptable carrier, adjuvant, or vehicle.

The amount of both the compound of the invention and the additional therapeutic agent (in those compositions containing the additional therapeutic agent described above) that can be combined with carrier substances to form a single dosage form will vary depending on the host being treated and specific route of administration. Preferably, the compositions of the present invention should be formulated such that a dose of a compound of the present invention ranging from 0.01 to 100 mg/kg body weight per day can be administered.

In those compositions that contain an additional therapeutic agent, the additional therapeutic agent and the compound of the present invention may act synergistically. Therefore, the amount of additional therapeutic agent in such compositions will be less than the amount required for monotherapy using only that therapeutic agent. In such compositions, a dose of the additional therapeutic agent may be administered ranging from about 0.001 to about 100 mg/kg body weight per day, or a dose of the additional therapeutic agent may be administered ranging from about 0.001 mg/kg to about 500 μg/kg, or from about 0.005 mg/kg to about 250 μg/kg, or about 0.01 mg/kg to about 100 μg/kg body weight per day.

The amount of additional therapeutic agent present in the compositions of the present invention will be no more than the amount that is typically administered in a composition containing that therapeutic agent as the sole active agent. Preferably, the amount of additional therapeutic agent in the presently disclosed compositions will be from about 50% to 100% of the amount typically present in a composition containing that agent as the sole therapeutically active agent.

In one embodiment, the present invention provides a composition comprising a compound of the present invention and one or more additional therapeutic agents. Therapeutic agents may be administered concomitantly with a compound of the present invention, or may be administered before or after administration of a compound of the present invention. Suitable therapeutic agents are described in more detail below. In some embodiments, a compound of the present invention may be administered prior to administration of the therapeutic agent at intervals of up to 5 minutes, 10 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 o'clock, 8 o'clock, 9 o'clock, 10 o'clock, 11 o'clock, 12 o'clock, 13 o'clock, 14 o'clock,

- 28 046380 hours, 16 hours, 17 hours or 18 hours. In other embodiments, a compound of the present invention may be administered after administration of the therapeutic agent at intervals of up to 5 minutes, 10 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 o'clock, 8 o'clock, 9 o'clock, 10 o'clock, 11 o'clock, 12 o'clock, 13 o'clock, 14 o'clock, 15 o'clock, 16 o'clock, 17 o'clock or 18 o'clock.

In some embodiments, the present invention provides a medicament comprising at least one compound of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

All features of each aspect of the present invention apply to all other aspects mutatis mutandis.

In order that the invention described herein may be more clearly understood, the following examples are given. It should be understood that these examples are for illustrative purposes only and should not be construed as limiting the present invention in any way.

Illustrative Example

As shown in the examples below, in some specific example embodiments, compounds were prepared in accordance with the following general procedures. It should be understood that while the general methods depict the synthesis of certain compounds of the present invention, the following general methods and other methods known to one skilled in the art can be applied to all compounds and subclasses and species of each of these compounds as described herein .

General procedures

X-ray powder diffraction (XRPD) analysis was performed using a Rigaku Smart-Lab X-ray diffraction system. The Rigaku SmartLab X-ray diffraction system was configured for Bragg-Brentano reflection geometry using a line source X-ray beam. The X-ray source was a long thin focus copper (Cu) tube operated at 40 kV and 44 mA. This source provides an incident beam profile on the sample that varies from a narrow line at high angles to a wide rectangle at low angles. A linear X-ray source uses slits to control the beams to ensure that the maximum beam size is less than 10 mm both along the line and perpendicular to the line. The Bragg-Brentano geometry is a parafocusing geometry controlled by passive divergence and receiving slits, with the sample itself acting as the focusing component for the optics. The resolution of the Bragg-Brentano geometry is determined in part by the radius of the diffractometer and the width of the reception slit used. Typically, Rigaku Smart-Lab is used to obtain peaks with a width of 0.1°2θ or less. The axial divergence of the X-ray beam is controlled using 5.0° Soller slits in both the incident and diffracted beam paths. The device is qualified using the American Society for Testing and Materials (ASTM) silicon standard on the day of analysis.

Powdered samples were prepared in a low-background silicon (Si) holder using light finger pressure to keep the surface of the samples flat and flush with the reference surface of the sample holder. Each sample was analyzed over a range of 2θ angles from 2 to 40° using a continuous scan of 6°2θ per minute with an effective step size of 0.02°2θ.

Thermogravimetric analysis (TGA) was carried out using a TA Instruments Q50 instrument. The scales of this instrument were calibrated using class M weights and temperature calibration was performed using alumel. Nitrogen purge was ~40 ml per minute in the scale area and ~60 ml per minute in the oven area. Each sample (approximately 2-5 mg) was placed in a pre-tared platinum crucible and heated from 20°C to 350°C at a rate of 10°C per minute. The heating rate may affect the analysis results. Nitrogen purge rates may vary depending on specific instrument specifications.

Differential scanning calorimetry (DSC) analyzes were performed using a TA Instruments Q2000. Temperature calibration of this device was performed using indium. During each analysis, the DSC cell was purged with nitrogen at a rate of ~50 ml per minute. The sample (approximately 1-2 mg) was placed in a standard rolled aluminum crucible and heated from 25°C to 350°C at a rate of 10°C per minute. The type of crucible, the preparation of the crucible for analysis, and the heating rate can affect the results of the analysis. Nitrogen purge rates may vary depending on specific instrument specifications.

Dynamic vapor sorption (DVA) analysis was performed using a TA Instruments Q5000 dynamic vapor sorption analyzer. This instrument was calibrated using standard weights and a humidity standard of sodium bromide. Approximately 20 mg of sample was added to a metal-lined quartz crucible for analysis. The specified sample was subjected to analysis at 25°C with a maximum equilibrium time of one hour, in increments

- 29 046380 relative humidity (r.h.) 10% in the range from 5 to 95% r.h. ow. (adsorption cycle) and from 95 to 5% rel. ow. (desorption cycle). The transition from one step to the next occurred either after meeting the equilibrium criterion, which was 0.01% change in mass, or, if the specified equilibrium criterion was not met, after one hour. Percentage weight change values were calculated using Microsoft Excel®. Temperature during EAF analysis can affect the results.

Karl Fischer (KF) analysis was performed using a Mettler-Toledo C20 coulometric KF titrator. This instrument was calibrated using a Hydranal water standard containing 1% water. The titrant was a methanol solution of Hydranal. The sample was analyzed in triplicate.

Optical microscopy (OM) analysis was performed on a Leica DM 2500 P compound microscope with a 10× eyepiece and a 10× objective for a total magnification of 100×. Images were acquired using a Qlmaging MicroPublisher 3.3 RTV camera. The polarization microscopy image (in color) was obtained using the sample in mineral oil with transmitted light from the microscope and polarizers in working position.

Infrared (IR) spectra were obtained using a Thermo Nicolet Model 6700 Fourier Transform (FTIR) spectrophotometer equipped with a deuterated triglycine sulfate (DTGS) detector, a potassium bromide (KBr) beam splitter, and an IR light source with electronic temperature control ( ECT) Ever-Glo®. The specified device was equipped with a SMART iTR attenuated total internal reflection (NOVO) sampling attachment with a diamond crystal. Single-beam scans of the background (air) and sample were obtained using 128 signal-averaged scans with a resolution of 2 cm ^-1 in the spectral range of 4000-400 cm ^-1 . The final spectrum of the sample was automatically calculated and presented in Log 1/R units. Wavelength calibration was verified using a certified polystyrene standard. Data collection and processing were carried out using Omnic 9.7.46 software.

Raman spectra with SF (Raman-SF spectra) were obtained on a Nicolet spectrometer, model 6700, coupled with an additional Nexus Raman module. This device is equipped with an Nd:YAG laser operating at a wavelength of 1024 nm, a CaF ₂ beam splitter and an indium gallium arsenide detector. Samples were packaged in a 3-inch glass NMR spectroscopy vial for analysis. The Raman-PF spectrum was obtained using 256 signal-averaged scans with a resolution of 4 cm ^-1 in the spectral range 3700-100 cm ^-1 . Data collection and processing were carried out using Omnic 9.7.46 software.

¹³ C nuclear magnetic resonance (NMR) spectra were obtained through ¹³ C cross-polarization and magic angle spinning (CPMAS) solid-state experiments performed on a 363 MHz Tecmag-based spectrometer. Each sample (approximately 200 mg) was packaged in a 7 mm zirconia rotor capped with polytrifluorochloroethylene (kel-F) end caps for subsequent data collection. Glycine, the value of which was taken to be 176.0 ppm, was used as an external standard. The data collection and processing parameters used are presented in Table 3 below.

Table 3

Data acquisition and processing parameters for ¹³ C NMR

Core ¹³ C Temperature (K) 293

- 30 046380

Observed Frequency (MHz) 91.37 Sweep Range (Hz) 29762 Time between sampling two adjacent points (µs) 33.6 1H pulse (µs) 5 Cross-polarization time (CP) (ms) 1 Contact time by Hartmann-Hahn (ms) 1 Pulse Width (µs) 1000 Accumulation time (ms) 34 Waiting time for magnetization recovery (s) 20 Rotation speed (kHz) 7.0 Number of scans 176 Processing options Reference external Line broadening (Hz) 15

Grinding was carried out using a Retsch Mill. Approximately 20 mg of material was placed in a plastic grinding cup, followed by the addition of 10 μL of water and a stainless steel ball. The sample was then crushed at 100% power for 20 minutes.

Example A - General Procedure for Preparation of Compound A

The title compound was prepared in accordance with the steps and intermediates (see, for example, Scheme 1 and Scheme 2) described below and in the '131 patent and the '363 publication, the contents of each and every one of which are incorporated herein by reference in their entirety. .

- 31 046380

Scheme 1 - Synthesis of compound A acetone, H ₂ SO ₄

D-Ribose --------but

93%

He

TBDPSCI, Et ₃ N, CH ₂ C1 ₂ TBDPSO

99%

TBDPS=tert-butyl diphenylsilyl

He

Ph ₃ PCH ₃ Br, KOBu', THF

93%

COCI ₂ , DMSO, CH ₂ CI ₂ , -78°C

RO

RO

96%

Vinyl MgBr, THF, -78°C

84% no.

ro——/ o' .o

Zhan cat-1 B, toluene, 60°C, 2 h

BUT.

80% ro- )—' o\ .0

O\ .0

NaBH ₄ , CeC1 _{3.7H 2} O, MeOH

84%

RO

CH ₂ I ₂

Et ₂ Zn, pQ

CH ₂ CI ₂ \

2,6-dichloropurine, Ph ₃ P, DIAD, THF, rt

RO

HE

O. .O

81% he is o. .O

75%

DIAD=diisopropyl azodicarboxylate

Zhang's catalyst Zhan cat-1B has the following structure:

Scheme 2 shows the remainder of the synthesis.

Scheme 2 - Synthesis of compound A (continued) CI.

RO

N

RO

N

N

P = TBDPS

NH ₂

N

'CI

80%

N

SMe 'SMe

90%

10% TFA, MeOH, 70 °C tube

75%

2M NH ₃ in isopropanol, sealed tube, °C PO

OH OH _l , 14 but'

PDC, MS, DMF

84%

PDC=pyridinium dichromate

C>O '01

MeSNa, DMF, 90 °C

Example 1 - Preparation of Compound A Free Base Form A

Compound A was prepared in accordance with the methods described in detail in Example 9 of the '131 patent and the '363 publication, the contents of each or each of which are incorporated herein by reference in their entirety. Scheme 3 below provides details of the synthesis of Compound A. The intermediate from the synthesis shown in Scheme 3 was used to prepare Form A of Compound A.

- 32 046380

Scheme 3 - Synthesis of compound A

Form A of Compound A

Compound A Form A was prepared as described below.

Form A of Compound A - Preparation Method 1

Compound 13 according to Scheme 3 (350 g, 0.58 mol) and water (2 L) were added to a 3 L round bottom flask. 200 ml of trifluoroacetic acid (TFA) was added and the resulting mixture was stirred at 60°C for 12 hours. HPLC indicated that the reaction was complete. The mixture was cooled to room temperature, washed with methylene chloride (MX, 300 ml * 3) and concentrated. Amberlyst 21 base resin was added to the resulting mixture to achieve a pH of approximately 9, and the resulting mixture was mixed until

- 33 046380 16 hours at room temperature. The mixture was then filtered, washed with methanol (MeOH) and concentrated. Purify by column chromatography [SiO2 (5X), MX/MeOH = 15/1 to 5/1 (100 vol)] to give Compound A as a light yellow solid. Compound A was then dissolved in HPLC grade MeOH to a clear solution, and the resulting solution was evaporated on a rotary evaporator (water pump, bath temperature 35 °C). Form A of compound A was gradually formed during the concentration process. After most of the MeOH had been removed (until there was no change in mass), deionized water was added three times and the resulting solution was evaporated to dryness using a rotary evaporator (oil pump, bath temperature 35 °C) to remove the remaining MeOH. The resulting compound was further dried by rotary evaporation (oil pump, bath temperature 35°C) until no change in mass (~16 hours) to obtain Form A of compound A.

Form A of Compound A - Preparation Method 2

18.4 mg of compound A and 10 μl of water were added to a plastic grinding cup. A stainless steel ball was added. The sample was then ground in a Retsch mill at 100% power for 20 minutes at room temperature.

Form A of Compound A - Preparation Method 3

200.0 mg of compound A and 3.0 ml of water were added to a 20 ml glass vial. A magnetic stir bar was added and the vial was capped. The vial was placed on a heating/stirring plate and the suspension was stirred magnetically for 7 days at ambient temperature. The vial was centrifuged for 10 minutes and the mother liquor was decanted. The vial was blown with dry air for 10 minutes to dry the solid. Then the bottle was placed in a vacuum desiccator for 2 hours to work at room temperature.

Form A of Compound A - Preparation Method 4

200.0 mg of compound A and 3.0 ml of water were added to a 20 ml glass vial. A magnetic stir bar was added and the vial was capped. The vial was placed on a heating/stirring plate set at 60°C and the suspension was stirred magnetically for 3 days. The vial was centrifuged for 10 minutes and the mother liquor was decanted. The vial was blown with dry air for 10 minutes to dry the solid. Then the bottle was placed in a vacuum desiccator for 2 hours to work at room temperature.

The above table 1 is shown below and shows the X-ray diffraction peaks observed for Form A of Compound A.

Table 1

XRPD Peak Positions for Form A of Compound A

°20 Relative intensity °20 Relative intensity °20 Relative intensity 7.6 1.49 21.5 35.63 30.8 12.79 8.0 100 22.8 2.30 32.0 8.32 9.0 3.49 23.1 6.16 32.8 8.27 10.8 4.60 23.7 4.21 32.9 13.11 11.8 1.59 23.9 9.39 33.7 9.73 12.5 4.59 24.9 54.39 34.7 4.31 13.1 59.02 26.1 32.89 36.3 3.98 16.2 34.56 26.5 13.26 36.7 8.82 16.7 37.68 26.6 22.22 37.9 12.68 17.2 10.65 27.1 60.62 38.2 3.16 17.9 45.59 28.6 10.52 38.5 1.75 18.1 16.05 29.3 1.66 38.7 2.83 18.3 10.91 29.7 9.05 39.6 2.37 19.8 4.96 30.1 1.89 — — 21.0 30.62 30.4 1.77 — —

In table Figure 4 below shows the IR-PF peaks observed for Form A of Compound A.

- 34 046380

Table 4

List of peaks in the IR-PF spectrum for Form A of compound A

Wave numbers (cm') 3460 1329 870 3293 1310 828 3178 1245 786 3114 1227 742 3008 1199 680 2921 1129 636 2887 1089 560 2871 1072 545 1629 1054 523 1580 1041 477 1510 1020 450 1457 975 418 1423 945 — 1406 886 —

In table Figure 5 below shows the peaks in the Raman spectrum observed for Form A of Compound A.

Table 5

List of peaks in the Raman-PF spectrum for Form A of compound A

Raman shift (cm ¹ ) 3085 1105 3052 1042 3009 975 2924 895 2902 847 2873 783 2826 742 1578 727 1512 715 1461 703 1447 628 1424 613 1341 564 1327 542 1251 — 1223 —

In table Figure 6 below shows the peaks in the ¹³ C NMR spectrum observed for Form A of Compound A.

- 35 046380

Table 6

List of peaks in the ¹³ C NMR spectrum for Form A of compound A

In table Figure 7 below shows selected ¹³ C NMR peaks and chemical shifts from the downfield peaks observed for Form A of Compound A.

Table 7 ¹³ C NMR peaks and chemical shifts for Form A of compound A

Peak positions in parts per million (ppm) A (ppm) from the lowest field peak 116.6 46 143.7 19 149.6 13 154.2 9 163.0

In fig. Figure 1 shows the XRPD diffraction pattern of Form A of Compound A.

In fig. Figure 2 shows the DSC curve of Form A of Compound A.

In fig. Figure 3 shows the TGA curve of Form A of Compound A.

In fig. Figure 4 shows the chipboard curve of Form A of connection A.

In fig. Figure 5 shows an XRPD diffraction pattern of Form A of Compound A before (top) and after (bottom) DSP analysis.

In fig. 6 shows the IR-PF spectra of Form A of compound A.

In fig. Figure 7 shows the Raman-PF spectra of Form A of compound A.

In fig. Figure 8 shows the solid-state ¹³ C spectra of Form A of compound A.

In fig. Figure 9 is an optical microscope image of Form A of Compound A.

Form A of Compound A was found to have the characteristics described below.

Based on the XRPD data, Form A of Compound A is crystalline and exhibits clear and well-resolved X-ray diffraction signals.

Based on optical microscopy images, Form A of Compound A appears as blade-shaped crystals of similar size and shape.

Form A of Compound A loses water of hydration and exhibits a melting point of approximately 196°C based on DSC data.

Based on the TG and CF data, Form A of Compound A is hydrated, containing 1.2 to 1.4 moles of water per mole of Form A of Compound A.

Form A of Compound A is stable under varying humidity conditions. It is moderately hygroscopic based on EAF data where a moisture gain of approximately 9.3% was observed during moisture sorption and a moisture loss of approximately 9.3% was observed during the desorption cycle.

- 36 046380 shaya approximately 2%. The crystalline form remained unchanged after EAF analysis where the sample was exposed to relative humidity up to 95% RH. ow. and reduced humidity to 5% rel. ow. Form A of Compound A is stable when crushed in the presence of water or when stirred in water at temperatures ranging from room temperature to 60°C. Form A of Compound A is more stable than Form B in solvent systems having a water activity greater than or equal to 0.72. If the solvent system has a water activity greater than or equal to 0.72 and the two crystalline forms are mixed, the mixture becomes Form A. These solvent systems can be a mixture of any organic solvents and organic solvents containing water.

Form A of Compound A has a solid-state ¹³ C NMR spectrum containing single ¹³ C signals for each carbon position in the chemical structure. Thus, there is one molecule in the asymmetric unit of crystal form A.

Based on the foregoing, Form A of Compound A is a moderately hygroscopic, stable, hydrated, crystalline material.

Example 2 - Preparation of Compound A Free Base Form B

HE

Compound A

Form B of compound A

Compound A Form B was prepared as described below.

Form B of compound A - preparation method 1

To a 20 ml glass vial, 200.4 mg of Compound A Form A and 3.0 ml of acetone were added. A magnetic stir bar was added and the vial was capped. The vial was placed on a heating/stirring plate set at 60°C and the suspension was stirred magnetically for 3 days. The vial was centrifuged for 10 minutes and the mother liquor was decanted. The vial was blown with dry air for 10 minutes to dry the solid. Then the bottle was placed in a vacuum desiccator for 2 hours to work at room temperature.

Form B of compound A - preparation method 2

17.2 mg Form A of Compound A and 1 ml ethyl acetate were added to a 1 dram glass vial. The resulting suspension was stirred magnetically on a heating/stirring plate set at 60° C. and ethanol (absolute) was added until dissolution occurred (1 ml was added). The magnetic stirrer armature was removed, the vial was capped, and the heating was turned off. After cooling to ambient temperature, no solid was detected and the vial was left overnight at ambient temperature, during which time no crystallization occurred. The vial was then placed in a refrigerator at approximately 5°C and left for 3 days, during which time no crystallization occurred. The vial was then placed in a freezer at approximately -15°C and left for 2 days, during which time crystallization occurred. The solid was isolated by filtration.

Form B of Compound A - Preparation Method 3

18.4 mg of Form A of Compound A and 10 μl of ethanol were added to a plastic grinding cup. A stainless steel ball was added. The sample was then ground in a Retsch mill at 100% power for 20 minutes.

Form B of compound A - preparation method 4

3.8 mg of Form A of Compound A and 0.7 ml of tetrahydrofuran were added to a 1 gram glass vial. The solid dissolved. The contents of the vial were then placed into a 20 ml glass vial containing 2 ml of hexanes. The 20 ml vial was stoppered and the sample was left for 6 days at ambient temperature during which crystallization occurred. The solvent was decanted and the solids were allowed to air dry.

The above table 2 is shown below and shows the X-ray diffraction peaks observed for Form B of Compound A.

- 37 046380

table 2

XRPD Peak Positions for Form B of Compound A

°20 Relative intensity °20 Relative intensity °20 Relative intensity 4.7 2.43 20.1 88.35 29.5 4.27 7.6 3.96 21.0 42.70 29.9 3.19 9.5 14.73 21.5 68.69 30.2 6.20 10.0 10.55 23.0 2.13 30.6 7.95 10.5 7.20 23.8 53.95 31.6 4.20 13.8 20.73 24.3 8.09 32.3 1.42 14.2 10.53 24.6 3.80 32.7 5.03 14.7 40.39 25.4 5.74 33.1 4.67 15.2 6.59 25.6 8.51 33.6 3.56 15.4 14.70 25.9 35.31 35.9 7,I 16.2 4.80 26.2 20.76 37.0 2.87 17.1 24.58 26.6 16.53 37.4 1.78 17.9 58.03 27.6 7.29 39.0 1.26 18.3 12.94 28.8 25.11 — — 19.0 100 29.1 8.40 — —

In table Figure 8 below shows the IR-PF peaks observed for Form B of Compound A.

Table 8

List of peaks in the IR-PF spectrum for Form B of compound A

Wave numbers (cm ¹ ) 3307 1335 835 3248 1298 791 3172 1239 750 3114 1205 721 2898 1190 694 2867 1171 668 1667 1120 639 1589 1093 613 1573 1074 607 1502 1042 574 1443 1023 553 1432 999 499 1409 970 436 1379 932 427 1368 884 418 1347 858 401

In table Figure 9 below shows the peaks in the Raman spectrum observed for Form B of Compound A.

- 38 046380

Table 9

List of peaks in the Raman-PF spectrum for Form B of compound A.

Raman shift (cm') 3511 997 3424 931 3115 837 3036 788 2931 750 2900 715 2731 669 1570 641 1502 613 1455 546 1371 446 1337 433 1319 446 1288 403 1248 306 1207 183 1190 160 1039 —

In table 10 below shows the peaks in the ¹³ C NMR spectrum observed for Form B of Compound A.

Table 10

List of peaks in the ¹³ C NMR spectrum for Form B of compound A

In table 11 below shows selected ¹³ C NMR peaks and chemical shifts from the downfield peaks observed for Form B of Compound A.

- 39 046380

Table 11 ¹³ C NMR peaks and chemical shifts for Form B of compound A

Peak positions in parts per million (ppm) A (ppm) from the lowest field peak 116.7 49 136.3 29 137.9 27 150.4 15 155.3 10 165.3

In fig. 10 shows the XRPD diffraction pattern of Form B of compound A.

In fig. Figure 11 shows the DSC curve of Form B of compound A.

In fig. 12 shows the TGA curve of Form B of Compound A.

In fig. 13 shows the chipboard curve of Form B of connection A.

In fig. 14 shows an XRPD diffraction pattern of Form B of Compound A before (top) and after (bottom) DSP analysis.

In fig. 15 shows the IR-PF spectra of Form B of compound A.

In fig. 16 shows the Raman-PF spectra of Form B of compound A.

In fig. Figure 17 shows the solid-state ¹³ C spectra of Form B of compound A.

In fig. 18 is an optical microscope image of Form B of Compound A.

Form B of Compound A was found to have the characteristics described below.

Based on the XRPD data, Form B of Compound A is crystalline and exhibits clear and well-resolved X-ray diffraction signals.

Based on optical microscopy images, Form B of Compound A appears as irregular crystals of similar size and shape.

Form B of Compound A exhibits a melting point of approximately 196°C based on DSC data.

Based on the TG and CF data, Form B of Compound A is anhydrous and unsolvated.

Form B of Compound A is moderately hygroscopic based on chipboard data where an increase in moisture of approximately 6.4% was observed during moisture sorption. During the desorption cycle, Form B of Compound A lost all of the resulting moisture. Additionally, the crystalline form remained unchanged after EAF analysis where the sample was exposed to relative humidity up to 95% RH. ow. and reduced to 5% rel. ow.

Form B of Compound A is more stable than Form A when suspended in organic/aqueous solvent systems having an aqueous activity of no more than 0.63. These solvent systems can be a mixture of any organic solvents and organic solvents containing water.

Form B of Compound A has a solid-state ¹³ C NMR spectrum containing some peaks that clearly appear to be two separate peaks. Thus, there are two conformationally distinct molecules in the asymmetric unit of crystalline form B.

Based on the foregoing, Form B of Compound A is a moderately hygroscopic, stable, unsolvated, anhydrous crystalline material.

Example 3 - Studies of single crystals of compound A

He

Compound A

Diffraction data for determining the structure of compound A are obtained by analysis using a cryo-electron microscope (Cryo-TEM) using microelectron diffraction (microED) techniques. microED techniques have been described in Jones et al., ACS Central Science 2018, 4, 1587-1592; Gruene et al., Angew. Chem. Int. Ed. 2018, 57, 16313-16317; and Shi etal, eLife 2013, 2, e01345, doi: 10.7554/eLife.01345.

-

Claims (41)

Materials and methods Compound A is prepared according to Example A described above without further crystallization or chemical modification other than crushing, grinding, grinding, grinding or other means of reducing particle size. To prepare Compound A samples for microED, approximately 1 mg of Compound A is placed between two microscope slides and ground to a fine powder. The ground powder is placed in an Eppendorf tube along with a transmission electron microscopy (TEM) grid and shaken. The loaded TEM grid is then removed from the Eppendorf tube and the said grid is gently tapped on filter paper to remove excess powder. After preparing grids containing samples of Compound A, they are then immersed in liquid nitrogen, placed in a sample cartridge, and introduced into a microscope for analysis. The copper grid with the perforated carbon film is cooled to liquid nitrogen temperatures and transferred to a cryoelectron microscope operating at an accelerating voltage of 200 kV (Thermo Fisher Talos Arctica). Diffraction data for 140 degrees is collected from the mononanocrystal by continuous rotation at 0.5 degrees per second. The complete data set is collected as a movie using a CetaD bottom-mount CMOS detector equipped with a thick scintillator layer for diffraction studies. Software written to convert film frames to SigmaTel Motion Video (SMV) format provides processing capability in X-ray detector software (XDS). Although the authors of the present description have described a number of embodiments of the present invention, it is obvious that the basic examples of the present invention can be modified to obtain other embodiments that use the compounds and methods of the present invention. Thus, it is to be understood that the scope of the present invention is to be defined by the appended claims and not by the specific embodiments which have been presented by way of example. CLAIM 1. Solid form of compound A: wherein said solid form is characterized by 3 or more peaks in the XRPD diffractogram selected from those at the following °2θ (degrees 2-theta) values: 7.6, 8.0, 9.0, 10.8, 11.8, 12, 5, 13.1, 16.2, 16.7, 17.2, 17.9, 18.1, 18.3, 19.8, 21.0, 21.5, 22.8, 23.1, 23.7, 23.9, 24.9, 26.1, 26.5, 26.6, 27.1, 28.6, 29.3, 29.7, 30.1, 30.4, 30, 8, 32.0, 32.8, 32.9, 33.7, 34.7, 36.3, 36.7, 37.9, 38.2, 38.5, 38.7 and 39.6, in this case, each value of °2θ is within ± 0.2 of the specified value. 2. Solid form of compound A: wherein said solid form is characterized by 3 or more peaks in the XRPD diffractogram selected from those at the following values of °2θ (degrees 2-theta): 4.7, 7.6, 9.5, 10.0, 10.5, 13, 8, 14.2, 14.7, 15.2, 15.4, 16.2, 17.1, 17.9, 18.3, 19.0, 20.1, 21.0, 21.5, 23.0, 23.8, 24.3, 24.6, 25.4, 25.6, 25.9, 26.2, 26.6, 27.6, 28.8, 29.1, 29, 5, 29.9, 30.2, 30.6, 31.6, 32.3, 32.7, 33.1, 33.6, 35.9, 37.0, 37.4 and 39.0, in this case, each value of °2θ is within ± 0.2 of the specified value. 3. Solid form according to claim 1 or 2, wherein said solid form is crystalline. 4. Solid form according to claim 1 or 2, wherein said solid form is a crystalline solid and does not contain amorphous compound A. 5. Solid form according to any one of claims 1 to 4, where said solid form does not contain impurities. 6. The solid form of claim 1, characterized by one or more peaks in an X-ray powder diffraction (XRPD) pattern selected from the peaks at 8.0 and 13.1 degrees 2-theta, wherein - 41 046380 each °2θ value is within ±0.2 of the specified value. 7. The solid form of claim 6, characterized by two peaks in the XRPD diffractogram selected from the peaks at 8.0 and 13.1 degrees 2-theta, each °2θ value being within ±0.2 of the indicated value. 8. A solid form according to claim 1, wherein said solid form is Form A. 9. The solid form according to claim 1, characterized by an XRPD diffraction pattern corresponding to the diffraction pattern shown in FIG. 1. 10. The solid form of claim 2, characterized by one or more peaks in the XRPD diffractogram selected from peaks at 9.5, 10.5 and 13.8 degrees 2-theta, each °2θ value being within ±0. 2 from the specified value. 11. The solid form of claim 10, characterized by at least two peaks in the XRPD diffractogram selected from peaks at 9.5, 10.5 and 13.8 degrees 2-theta, each °2θ value being within ± 0.2 of the specified value. 12. The solid form according to claim 11, wherein said solid form is Form B. 13. The solid form of claim 2, characterized by an XRPD diffraction pattern corresponding to the diffraction pattern shown in FIG. 10. 14. The solid form of claim 10, characterized by peaks in the XRPD diffractogram at 9.5, 10.5 and 13.8 degrees 2-theta, with each °2θ value being within ±0.2 of the indicated value. 15. The solid form of claim 1, characterized by 5 or more peaks in the XRPD diffractogram selected from those at the following °2θ (degrees 2-theta): 7.6, 8.0, 9.0, 10.8, 11 ,8, 12.5, 13.1, 16.2, 16.7, 17.2, 17.9, 18.1, 18.3, 19.8, 21.0, 21.5, 22.8 , 23.1, 23.7, 23.9, 24.9, 26.1, 26.5, 26.6, 27.1, 28.6, 29.3, 29.7, 30.1, 30 ,4, 30.8, 32.0, 32.8, 32.9, 33.7, 34.7, 36.3, 36.7, 37.9, 38.2, 38.5, 38.7 and 39.6, with each °2θ value being within ±0.2 of the specified value. 16. The solid form of claim 15, characterized by one or more peaks in the XRPD diffractogram selected from 8.0 ± 0.2 and 13.1 ± 0.2 degrees 2-theta. 17. The solid form of any one of claims 1, 15 or 16, characterized by one or more peaks in the XRPD diffractogram selected from 24.9 ± 0.2 and 27.1 ± 0.2 degrees 2-theta. 18. Solid form according to any one of claims 1 or 15-17, containing at least 95% by weight of crystalline compound A and free of impurities. 19. Solid form according to any one of claims 1 or 15-18, characterized in that said solid form is hydrated. 20. Solid form according to any one of claims 1 or 15-19, containing from 1.2 to 1.4 moles of water per mole of compound A. 21. The solid form of claim 1, having a solid ¹³ C spectroscopy spectrum containing one or more of the following peaks, in parts per million (ppm): 12.8, 15.0, 20.4, 33.9, 60, 4, 66.7, 71.8, 75.4, 116.6, 143.7, 149.6, 154.2 and 163.0, while the specified spectrum contains peaks at 20.4, 33.9, 71 .8, 143.7 and 163.0. 22. The solid form according to claim 1, having a solid-state ¹³ C spectroscopy spectrum corresponding to the spectrum shown in FIG. 8. 23. Solid form according to claim 1, having a Raman-PF spectrum containing one or more of the following peaks, cm ^-1 : 3085, 3052, 3009, 2924, 2902, 2873, 2826, 1578, 1512, 1461, 1447 , 1424, 1341, 1327, 1251, 1223, 1105, 1042, 975, 895, 847, 783, 742, 727, 715, 703, 628, 613, 564 and 542, with the specified spectrum containing peaks at 1578 and 1512 . 24. The solid form according to claim 1, having a Raman-PF spectrum corresponding to the spectrum shown in FIG. 7. 25. Solid form according to claim 1, having an IR-PF spectrum containing one or more of the following peaks, cm ^-1 : 3460, 3293, 3178, 3114, 3008, 2921, 2887, 2871, 1629, 1580, 1510, 1457, 1423, 1406, 1329, 1310, 1245, 1227, 1199, 1129, 1089, 1072, 1054, 1041, 1020, 975, 945, 886, 870, 828, 786, 742, 680, 636, 560, 545, 523, 477, 450 and 418, with this spectrum containing a peak at 1629. 26. Solid form according to claim 1, having an IR-PF spectrum corresponding to the spectrum shown in FIG. 6. 27. The solid form of claim 2, characterized by 5 or more peaks in the XRPD diffractogram selected from those at the following °2θ (degrees 2-theta): 4.7, 7.6, 9.5, 10.0, 10 .5, 13.8, 14.2, 14.7, 15.2, 15.4, 16.2, 17.1, 17.9, 18.3, 19.0, 20.1, 21.0 , 21.5, 23.0, 23.8, 24.3, 24.6, 25.4, 25.6, 25.9, 26.2, 26.6, 27.6, 28.8, 29 ,1, 29.5, 29.9, 30.2, 30.6, 31.6, 32.3, 32.7, 33.1, 33.6, 35.9, 37.0, 37.4 and 39.0, with each °2θ value being within ±0.2 of the specified value. 28. The solid form according to any one of claims 2 or 27, characterized by one or more peaks in the XRPD diffractogram selected from peaks at 14.2 ± 0.2, 14.7 ± 0.2, 15.2 ± 0.2, 15.4 ± 0.2 and 19.0 ± 0.2 degrees 2-theta. 29. Solid form according to any one of claims 2, 27 or 28, characterized by two or more peaks in the XRPD diffractogram selected from peaks at 14.2 ± 0.2, 14.7 ± 0.2, 15.2 ± 0, 2, 15.4 ± 0.2 and 19.0 ± 0.2 - 42 046380 degrees 2-theta. 30. The solid form according to any one of claims 2 or 27-29, characterized by peaks in the XRPD diffractogram at 14.7 ± 0.2 and 19.0 ± 0.2 degrees 2-theta. 31. Solid form according to any one of claims 2 or 27-30, characterized by one or more peaks in the XRPD diffractogram selected from peaks at 9.5 ± 0.2, 10.5 ± 0.2 and 13.8 ± 0, 2 degrees 2-theta. 32. The solid form according to any one of claims 2, 27 or 28, characterized by XRPD peaks at 9.5 ± 0.2, 10.5 ± 0.2 and 13.8 ± 0.2 degrees 2-theta. 33. Solid form according to any one of claims 2 or 27-32, containing at least 95% by weight of crystalline compound A and free of impurities. 34. Solid form according to any one of claims 2 or 27-33, characterized in that said solid form is anhydrous and unsolvated. 35. The solid form of claim 2, having a solid ¹³ C spectroscopy spectrum containing one or more of the following peaks, in parts per million (ppm): 12.8, 23.2, 36.0, 37.5, 61. 9, 63.8, 68.2, 73.9, 76.4, 77.4, 116.7, 136.3, 137.9, 150.4, 155.3, and 165.3, with the specified the spectrum contains peaks at 23.2, 63.8, 73.9, 77.4, 136.3, 137.9 and 165.3. 36. The solid form according to claim 2, having a solid-state ¹³ C spectroscopy spectrum corresponding to the spectrum shown in FIG. 17. 37. Solid form according to claim 2, having a Raman-PF spectrum containing one or more of the following peaks, cm ^-1 : 3511, 3424, 3115, 3036, 2931, 2900, 2731, 1570, 1502, 1455, 1371, 1337, 1319, 1288, 1248, 1207, 1190, 1039, 997, 931, 837, 788, 750, 715, 669, 641, 613, 546, 446, 433, 446, 403, 306, 183 and 1 60, while the indicated spectrum contains peaks at 1570 and 1502. 38. The solid form according to claim 2, having a Raman-PF spectrum corresponding to the spectrum shown in FIG. 16. 39. Solid form according to claim 2, having an IR-PF spectrum containing one or more of the following peaks, cm ^-1 : 3307, 3248, 3172, 3114, 2898, 2867, 1667, 1589, 1573, 1502, 1443, 1432, 1409, 1379, 1368, 1347, 1335, 1298, 1239, 1205, 1190, 1171, 1120, 1093, 1074, 1042, 1023, 999, 970, 932, 884, 858, 83 5, 791, 750, 721, 694, 668, 639, 613, 607, 574, 553, 499, 436, 427, 418 and 401, with the spectrum containing a peak at 1667. 40. The solid form according to claim 2, having an IR-PF spectrum corresponding to the spectrum shown in FIG. 15. 41. A pharmaceutical composition containing a solid form according to any one of claims 1 to 40 and a pharmaceutically acceptable carrier, excipient or adjuvant, for the treatment of traumatic brain injury (TBI), stroke, neurodegenerative condition, heart disease or cardiovascular disease, concussion, partial or complete transection of the spinal cord, malnutrition, toxic neuropathies, meningoencephalopathies, neurodegeneration caused by hereditary disease, age-related neurodegeneration, vascular disease, Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), multiple sclerosis (MS) , amyotrophic lateral sclerosis (ALS), chronic traumatic encephalopathy (CTE), autoimmune diseases, allergic diseases, transplant rejection, graft-versus-host disease, intraocular hypertension, glaucoma, odor sensitivity, smell impairment, type 2 diabetes and/or pain management, respiratory diseases, central nervous system dysfunction, learning disabilities, cognitive impairment, hearing impairment, Meniere's disease, endolymphatic hydrops, progressive hearing loss, dizziness, vertigo, tinnitus, concomitant brain damage associated with radiation therapy for cancer, migraine treatment, sleep disorders in the elderly, epilepsy, schizophrenia, symptoms occurring in recovering alcoholics, neuronal or peripheral nervous system damage during surgery, gastrointestinal disorders, CNS-mediated pain, migraines, concomitant brain damage associated with radiation therapy cancer, depression, changes in mood or behavior, dementia, erratic behavior, suicidal tendencies, tremor, Huntington's chorea, decreased motor coordination, hearing loss, speech disorders, dry eye syndrome, hypomimia, attention deficit, memory loss, cognitive impairment, dysarthria, dysphagia , visual disturbances or disorientation; a condition associated with brain damage or a neurodegenerative condition; neuropathic pain, inflammatory pain or acute pain; or to enhance cardioprotection or regeneration of damaged cardiac tissue in a patient in need who has suffered cardiac ischemia or myocardial infarction; or enhancing neuroprotection or restoration of neural tissue function in a patient in need who has suffered a TBI or stroke; or to prevent, alleviate, treat, or promote recovery from addiction, addictive behavior, behavioral addiction, brain reward system disorder, compulsive disorder, or related condition. -