Solid forms of spinous formation promoter

Abstract

Provided herein are solid forms of a spinous formation promoter. Provided herein are crystalline forms of 2-(2-(2-(2-(4-(benzo [d] thiazol-2-yl) phenoxy) ethoxy) ethoxy) ethoxy) ethyl-1-ol (Compound I), as well as methods of making and methods of using the crystalline forms. # imgabs0 #

Description

[0001] This application is a divisional application of the Chinese invention patent application with application date of January 30, 2020, application number 202080025035.4, and invention name “Solid form of spines growth promoter”.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims the benefit under 35 U.S.C. §119(e) of U.S. Provisional Application No. 62 / 799,644, filed on January 31, 2019, which is incorporated herein by reference in its entirety. Technical Field

[0004] The present disclosure generally relates to crystalline forms of the compound 2-(2-(2-(2-(4-(benzo[d]thiazol-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)ethan-1-ol, referred to herein as Compound 1, their pharmaceutical compositions, their therapeutic uses, and methods of preparing these forms. Background Art

[0005] Neurological diseases are diseases of the brain, spinal cord and peripheral nervous system. In terms of epidemiology and individual morbidity, the greatest social cost is caused by neurodegenerative conditions, which can cause damage or loss of neurons and synaptic connections therebetween. Among these neurological diseases, the most prominent are Alzheimer's disease and Parkinson's disease. Other neurodegenerative conditions include age-related conditions (e.g., Parkinson's dementia, vascular dementia, amyotrophic lateral sclerosis), genetic syndromes (e.g., Down syndrome), injury-related conditions (e.g., traumatic brain injury, chronic traumatic encephalopathy) and conditions typically considered as purely mental in nature, such as schizophrenia and depression.

[0006] Researchers have classified hundreds of neurological diseases, such as brain tumors, epilepsy, Alzheimer's disease, Parkinson's disease and stroke, and conditions associated with old age, such as dementia. Some of these conditions are caused by the progressive loss of synapses (the junction between two different neurons) and the loss of neurons (neurodegeneration). Unfortunately, neurodegenerative diseases are almost completely resistant to treatment. Neurons in the brain communicate with each other by sending neurons to synapses to release chemicals (neurotransmitters), thereby changing the potential of the receiving neurons. The part of the neuron that releases neurotransmitters is the axon (the presynaptic side of the synapse), and the part of the synapse affected by the neurotransmitter is called the dendritic spine (the postsynaptic side of the synapse). The number, position and even shape changes of synaptic junctions are the basis of memory, learning, thinking and personality. The part of the brain called the hippocampus is closely related to the formation of memory, and in neurodegenerative diseases, it will suffer from the obvious loss of synapses and neurons. The development of novel approaches to restore spine density in the hippocampus may have important implications for the treatment of many neurodegenerative and developmental cognitive disorders.

[0007] Therefore, small molecules that promote spine formation have potential use in ameliorating cognitive deficits in neuronal diseases, including neurodegenerative diseases such as Alzheimer's disease. However, for the treatment of diseases that respond to the promotion of spine formation, including neuronal diseases, there is a need for highly pure single polymorphic forms of compounds that are effective and exhibit improved pharmacokinetic and / or pharmacodynamic characteristics. Summary of the invention

[0008] Compound I exhibits activity in promoting spine generation and is described, for example, in International Publication No. WO2019 / 028164 (February 7, 2019), which is incorporated herein by reference in its entirety. Compound I has the following formula:

[0009]

[0010] Compound I can be synthesized according to the methods described herein and the methods described in International Publication No. WO2019 / 028164 (February 7, 2019).

[0011] The present disclosure provides forms of Compound 1 and salts, cocrystals, hydrates, and solvates thereof. Also described herein are methods of preparing forms of Compound 1, pharmaceutical compositions comprising crystalline forms of Compound 1, and methods of using such forms and pharmaceutical compositions to treat diseases responsive to spinogenesis. In some embodiments, the disease is a neuronal disease.

[0012] Thus, one embodiment is crystalline 2-(2-(2-(2-(4-(benzo[d]thiazol-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)ethan-1-ol (Compound 1 Form I) characterized by an X-ray powder diffraction pattern comprising the following peaks: about 4.6, about 20.8, and about 23.7 °2θ±0.2 °2θ, when measured using a wavelength of The Cu-Kα radiation was measured on a diffractometer.

[0013] Another embodiment is crystalline 2-(2-(2-(2-(4-(benzo[d]thiazol-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)ethan-1-ol (Compound I Form II) characterized by an X-ray powder diffraction pattern comprising peaks at about 9.2, about 13.8, and about 16.1 °2θ±0.2 °2θ, when a wavelength of The Cu-Kα radiation was measured on a diffractometer.

[0014] Another embodiment is crystalline 2-(2-(2-(2-(4-(Benzo[d]thiazol-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)ethan-1-ol (Compound I Form III) characterized by an X-ray powder diffraction pattern comprising peaks at about 7.7, about 10.3, and about 15.3 °2θ ± 0.2 °2θ, when a wavelength of The Cu-Kα radiation was measured on a diffractometer.

[0015] Another embodiment is crystalline 2-(2-(2-(2-(4-(Benzo[d]thiazol-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)ethan-1-ol (Compound I Form IV) characterized by an X-ray powder diffraction pattern comprising peaks at about 5.4, about 22.4, and about 23.3 °2θ ± 0.2 °2θ, as measured using a wavelength of The Cu-Kα radiation was measured on a diffractometer.

[0016] Another embodiment is crystalline 2-(2-(2-(2-(4-(Benzo[d]thiazol-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)ethan-1-ol (Compound I Form V) characterized by an X-ray powder diffraction pattern comprising peaks at about 4.6, about 9.2, and about 23.1 °2θ±0.2 °2θ, as measured using a wavelength of The Cu-Kα radiation was measured on a diffractometer.

[0017] Another embodiment is amorphous 2-(2-(2-(2-(4-(benzo[d]thiazol-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)ethan-1-ol.

[0018] Some embodiments provided herein relate to crystalline forms or co-crystals of a hydrate of Compound 1.

[0019] Another embodiment is directed to a pharmaceutical composition comprising one or more forms of Compound 1 as described herein and one or more pharmaceutically acceptable carriers.

[0020] Another embodiment is directed to the use of a Compound 1 form described herein or a pharmaceutical composition described herein for promoting spinogenesis in a patient in need thereof.

[0021] Another embodiment relates to a method of treating a disease responsive to spinogenesis in a patient in need thereof, comprising administering a therapeutically effective amount of a Compound I form as described herein or a pharmaceutical composition as described herein. In some embodiments, the disease is a neuronal disease. In some embodiments, the disease is selected from Alzheimer's disease, Parkinson's disease, Parkinson's dementia, autism, fragile X syndrome, and traumatic brain injury.

[0022] Another embodiment is directed to a method of treating Alzheimer's disease in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a Compound 1 form as described herein, or a pharmaceutical composition as described herein.

[0023] Another embodiment relates to the use of a Compound 1 form as described herein or a pharmaceutical composition as described herein for treating a disease selected from Alzheimer's disease, Parkinson's disease, Parkinson's dementia, autism, fragile X syndrome, and traumatic brain injury. The disease may be Alzheimer's disease.

[0024] Another embodiment relates to the use of a form of Compound 1 as described herein or a pharmaceutical composition as described herein for the preparation of a medicament for treating a disease selected from Alzheimer's disease, Parkinson's disease, Parkinson's dementia, autism, fragile X syndrome, and traumatic brain injury. The disease may be Alzheimer's disease.

[0025] In some embodiments, a method for preparing Compound 1 or a pharmaceutically acceptable salt thereof is provided:

[0026]

[0027]

[0028] Comprising contacting compound A with compound B under first reaction conditions comprising a halide to form compound I:

[0029]

[0030] In some embodiments, the method of preparing Compound 1 further comprises contacting Compound C with Compound D under second reaction conditions comprising a protic acid to form Compound A:

[0031]

[0032] In some embodiments, the method of preparing Compound I further comprises contacting Compound E with p-toluenesulfonyl chloride under third reaction conditions comprising a silver salt to form Compound B:

[0033]

[0034] The present invention also relates to the following items:

[0035] 1. Crystalline 2-(2-(2-(2-(4-(Benzo[d]thiazol-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)ethan-1-ol (Compound 1 Form I), characterized by an X-ray powder diffraction pattern comprising the following peaks (°2θ±0.2°2θ): about 4.6, about 20.8 and about 23.7, using a wavelength of The Cu-Kα radiation was measured on a diffractometer.

[0036] 2. The Compound 1 Form I according to claim 1, wherein the diffraction pattern further comprises peaks at about 9.2, about 16.3, about 18.6 and about 19.6 (°2θ±0.2°2θ).

[0037] 3. The Compound 1 Form I according to claim 1, characterized in that the differential scanning calorimetry (DSC) curve includes an endotherm at about 70°C.

[0038] 4. Crystalline 2-(2-(2-(2-(4-(Benzo[d]thiazol-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)ethan-1-ol (Compound I Form II), characterized by an X-ray powder diffraction pattern comprising the following peaks (°2θ±0.2°2θ): about 9.2, about 13.8 and about 16.1, using a wavelength of The Cu-Kα radiation was measured on a diffractometer.

[0039] 5. Compound I Form II according to item 4, wherein the diffraction pattern further comprises peaks at about 6.9, about 11.5 and about 18.4 (°2θ±0.2°2θ).

[0040] 6. Crystalline 2-(2-(2-(2-(4-(Benzo[d]thiazol-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)ethan-1-ol (Compound I Form III), characterized by an X-ray powder diffraction pattern comprising the following peaks (°2θ±0.2°2θ): about 7.7, about 10.3 and about 15.3, using a wavelength of The Cu-Kα radiation was measured on a diffractometer.

[0041] 7. Compound I Form III according to item 6, wherein the diffraction pattern further comprises peaks at about 5.1, about 12.8 and about 18.0 (°2θ±0.2°2θ).

[0042] 8. Crystalline 2-(2-(2-(2-(4-(Benzo[d]thiazol-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)ethan-1-ol (Compound I Form IV), characterized by an X-ray powder diffraction pattern comprising the following peaks (°2θ±0.2°2θ): about 5.4, about 22.4, and about 23.3, using a wavelength of The Cu-Kα radiation was measured on a diffractometer.

[0043] 9. Compound I Form IV according to item 8, wherein the diffraction pattern further comprises peaks at about 10.8, about 18.9, about 23.9 and about 26.8 (°2θ±0.2°2θ).

[0044] 10. Compound I Form IV according to item 8, characterized in that the differential scanning calorimetry (DSC) curve includes an endotherm at about 48°C and an endotherm at about 59°C.

[0045] 11. Crystalline 2-(2-(2-(2-(4-(Benzo[d]thiazol-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)ethan-1-ol (Compound I Form V), characterized by an X-ray powder diffraction pattern comprising the following peaks (°2θ±0.2°2θ): about 4.6, about 9.2, and about 23.1 at a wavelength of The Cu-Kα radiation was measured on a diffractometer.

[0046] 12. Compound I Form V according to item 11, wherein the diffraction pattern further includes peaks at about 13.8 and about 18.6 (°2θ±0.2°2θ).

[0047] 13. Compound I Form V according to item 11, characterized in that the differential scanning calorimetry (DSC) curve includes an endotherm at about 70°C.

[0048] 14. A pharmaceutical composition comprising one or more crystalline forms of any one of the preceding clauses and one or more pharmaceutically acceptable carriers.

[0049] 15. A method for promoting spine formation in a patient, comprising administering a therapeutically effective amount of the compound I form described in any one of items 1-13 or the pharmaceutical composition described in item 14.

[0050] 16. The method according to item 15, wherein the patient suffers from a neuronal disease.

[0051] 17. The method according to item 16, wherein the neuronal disease is selected from Alzheimer's disease, Parkinson's disease, Parkinson's dementia, autism, fragile X syndrome, depression and traumatic brain injury.

[0052] 18. The method according to item 17, wherein the neuronal disease is Alzheimer's disease.

[0053] 19. A method for preparing Compound I or a pharmaceutically acceptable salt thereof:

[0054]

[0055] Compound I

[0056] Comprising contacting compound A with compound B under first reaction conditions comprising a halide to form compound I:

[0057]

[0058] 20. The method according to item 19, wherein the halide is an alkali metal halide.

[0059] 21. The method according to item 20, wherein the alkali metal halide is an alkali metal iodide.

[0060] 22. The method according to item 21, wherein the alkali metal iodide is potassium iodide.

[0061] 23. The method according to item 19, wherein the first reaction condition further comprises an inorganic base.

[0062] 24. The method according to item 23, wherein the inorganic base is potassium carbonate.

[0063] 25. The method according to item 19, wherein the first reaction conditions comprise a temperature of 65 to 120°C.

[0064] 26. The method according to item 19, further comprising contacting compound C with compound D under second reaction conditions comprising a protic acid to form compound A:

[0065]

[0066] 27. The method according to item 26, wherein the protonic acid is an organic acid.

[0067] 28. The method according to item 27, wherein the organic acid is acetic acid.

[0068] 29. The method according to item 26, wherein the protic acid is present in a molar ratio greater than stoichiometric relative to compound C and compound D.

[0069] 30. The method according to item 29, wherein the protonic acid dissolves compound C and compound D.

[0070] 31. The method according to item 26, further comprising contacting compound E with p-toluenesulfonyl chloride under third reaction conditions comprising a silver salt to form compound B:

[0071]

[0072] 32. The method according to item 31, wherein the silver salt is Ag2O.

[0073] 33. The method according to item 31, wherein the third reaction condition further comprises an alkali metal iodide. BRIEF DESCRIPTION OF THE DRAWINGS

[0074] Figure 1 Shown is the X-ray powder diffraction (XRPD) of Compound 1 Form I.

[0075] Figure 2 Shown are the differential scanning calorimeter (DSC) curve and thermogravimetric analysis (TGA) of Compound 1 Form I.

[0076] Figure 3 Shown is the X-ray powder diffraction (XRPD) of Compound 1 Form II.

[0077] Figure 4 X-ray powder diffraction (XRPD) of Compound 1 Form III is shown.

[0078] Figure 5 X-ray powder diffraction (XRPD) of Compound 1 Form IV is shown.

[0079] Figure 6 Shown are the Differential Scanning Calorimeter (DSC) curve and Thermogravimetric Analysis (TGA) of Compound 1 Form IV.

[0080] Figure 7 X-ray powder diffraction (XRPD) of Compound 1 Form V is shown.

[0081] Figure 8 Shown are the differential scanning calorimeter (DSC) curve and thermogravimetric analysis (TGA) of Compound 1 Form V.

[0082] Fig. 9 The interconversion relationship of the crystalline forms of Compound I obtained during the polymorph screening process is shown.

[0083] Fig.10 An XRPD pattern of Compound 1 starting material is shown.

[0084] Fig.11 The TGA / DSC curve of Compound I starting material is shown.

[0085] Fig.12 A DVS pattern of the Compound 1 starting material is shown.

[0086] Fig.13 Shown is an XRPD overlay of Compound 1 starting material before and after DVS testing.

[0087] Fig.14 Shown is an XRPD overlay of Compound I after storage at 30°C / 65% RH / open.

[0088] Fig.15 Shown is an XRPD overlay of Compound I after storage at 30°C / 65% RH / closed.

[0089] Fig.16 Shown is an XRPD overlay of Compound I after storage at 40°C / 75% RH / open.

[0090] Fig.17 Shown is an XRPD overlay of Compound I after storage at 40°C / 75% RH / closed. DETAILED DESCRIPTION

[0091] The compound 2-(2-(2-(2-(4-(benzo[d]thiazol-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)ethan-1-ol, designated herein as Compound I, has the formula:

[0092]

[0093] The present disclosure relates to various crystalline forms of Compound 1, and methods of making these crystalline forms. Also described herein are forms of Compound 1 further labeled herein as "Compound 1 Form I," "Compound 1 Form II," "Compound 1 Form III," "Compound 1 Form IV," "Compound 1 Form V," and "amorphous Compound 1." In some embodiments, these forms of Compound 1 may be anhydrous. In some embodiments, these forms of Compound 1 may be solvates or hydrates.

[0094] Other crystalline forms of Compound 1 are further described herein. In some embodiments, the crystalline form of Compound 1 may include a salt or co-crystal of Compound 1.

[0095] definition

[0096] The following words and phrases as used in this specification are generally intended to have the meanings as set out below, unless otherwise indicated in the context in which they are used.

[0097] The term "fascin" refers to a 54-58 kDa protein that is an actin cross-linking protein. The term "fascin" may refer to the amino acid sequence of human fascin 1. The term "fascin" includes a nucleotide sequence or protein in wild-type form and any mutants thereof. In some embodiments, "fascin" is wild-type fascin. In some embodiments, "fascin" is one or more mutant forms. In some embodiments, fascin is human fascin 1. In some embodiments, fascin is encoded in the nucleotide sequence corresponding to reference number GI:347360903. In some embodiments, fascin is encoded in the nucleotide sequence of RefSeq M_003088. In some embodiments, the fascin corresponds to the amino acid sequence of RefSeq NP_003079.1.

[0098] The term "spine generation" and the like refer in the usual and customary sense to the development (e.g., growth and / or maturation) of dendritic spines of neurons. In some embodiments, the compounds provided herein promote spine generation without affecting spine morphology. Promotion is the absence relative to the administration of the compound.

[0099] As used herein, the term "dendrite" refers to a branch extension of a neuronal cell. Dendrites are generally responsible for receiving electrochemical signals transmitted from the axons of neighboring neurons. The term "dendritic spines" or "dendritic spines" refers to protoplasmic protrusions on a neuronal cell (e.g., a neuronal cell on a dendrite). In some embodiments, dendritic spines can be described as having a membranous neck, which can be terminated by a small head (e.g., head). Dendritic spines are classified according to their shape: headless, thin, stubby, mushroom or branch-like. Dendritic spine density refers to the total number of dendritic spines per unit length of a neuronal cell. For example, the dendritic spine density can be given as the number of dendritic spines per micron.

[0100] The term "dendritic spine formation" etc. refers to the process that causes the number of dendritic spines to increase or the development of dendritic spines to increase in the usual and customary sense. The term "dendritic spine morphology" etc. refers to the physical characteristics (e.g., shape and structure) of dendritic spines in the usual and customary sense. The improvement of dendritic spine morphology is a morphological change (e.g., an increase in length or an increase in width), which can lead to functional enhancement (e.g., an increase in the number of contacts between neurons or a decrease in the space (e.g., synaptic cleft) between adjacent neurons). As known in the art and disclosed herein, an exemplary method for this characterization includes measuring the size (i.e., length and width) of dendritic spines. Therefore, the term "improving dendritic spine morphology" generally refers to increasing the length, width, or both the length and width of dendritic spines.

[0101] "Binding" refers to at least two different species (e.g., chemical compounds comprising biomolecules or cells) coming into close enough proximity to react or interact, resulting in the formation of a complex. For example, the binding of two different species (e.g., a protein and a compound described herein) may result in the formation of a complex, wherein the species interact through non-covalent or covalent bonds. In some embodiments, the resulting complex is formed when two different species (e.g., a protein and a compound described herein) interact through non-covalent bonds (e.g., electrostatic, van der Waals, or hydrophobic).

[0102] As defined herein, the terms "activation," "activate," "activating," and the like with respect to a protein-activator (e.g., agonist) interaction mean actively affecting (e.g., increasing) the activity or function of a protein relative to the activity or function of the protein in the absence of the activator (e.g., a compound described herein).

[0103] "Control" or "controlled experiment" is used according to its ordinary general meaning and refers to an experiment in which the subjects or agents of the experiment are treated as in a parallel experiment, except that the experimental procedures, agents, or variables are omitted. In some instances, a control is used as a standard of comparison in evaluating the effects of an experiment.

[0104] "Contacting" is used according to its general ordinary meaning and refers to the process of bringing at least two different species (e.g., chemical compounds including biomolecules, or cells) into close enough proximity to interact. The term "contacting" can include allowing the two species to react or physically touch, where the two species can be compounds, biomolecules, proteins, or enzymes as described herein. In some embodiments, contacting includes allowing a compound described herein to interact with a protein (e.g., fascin) or an enzyme.

[0105] As defined herein, the terms "inhibition", "inhibit", "inhibiting" and the like will be given their customary meaning to those skilled in the art. With respect to protein-inhibitor (e.g., antagonist) interactions, the terms "inhibition", "inhibit", "inhibiting" mean negatively affecting (e.g., reducing) the functional activity of a protein relative to the functional activity of the protein in the absence of the inhibitor.

[0106] As used herein, the term "about" used in the context of quantitative measurements means ±10% of the indicated amount, or alternatively, ±5% or ±1% of the indicated amount.

[0107] The term "complex" refers to a formation resulting from the interaction between Compound I and another molecule.

[0108] The term "solvate" refers to a complex formed by combining Compound I with a solvent. As used herein, the term "solvate" includes hydrates (ie, solvates when the solvent is water).

[0109] The term "co-crystal" refers to a molecular complex of ionized or non-ionized Compound 1 (or any other form, salt or compound disclosed herein) and one or more non-ionized co-crystal formers (e.g., pharmaceutically acceptable salts) linked by non-covalent interactions. In certain embodiments, the co-crystal may have improved properties compared to the free form (i.e., free molecule, zwitterion, hydrate, solvate, etc.) or salt (which includes salt hydrates and solvates). In further embodiments, the improved properties are selected from: increased solubility, increased dissolution, increased bioavailability, increased dose response, reduced hygroscopicity, crystalline forms of normally amorphous compounds, crystalline forms of compounds that are difficult or non-salifiable, reduced form diversity, more desirable morphology, etc.

[0110] The term "co-crystal former" or "coformer" refers to one or more pharmaceutically acceptable bases and / or pharmaceutically acceptable acids disclosed herein in association with Compound 1 or any other compound disclosed herein.

[0111] Any chemical formula or structure given herein, including Compound I, is also intended to represent unlabeled forms of these compounds as well as isotopically labeled forms. An isotopically labeled compound has a structure depicted by the chemical formula given herein, except that one or more atoms are replaced by atoms having a selected atomic mass or mass number. Examples of isotopes that can be incorporated into the compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, such as, but not limited to: 2 H (deuterium, D), 3 H (tritium), 11 C. 13 C. 14 C. 15 N. 18 F. 31 P. 32 P. 35 S. 36 Cl, and 125 I. Various isotopically labeled compounds of the present disclosure can be prepared, for example, wherein an isotope (such as 3 H. 13 C and 14Such isotopically labeled compounds are useful in metabolic studies, reaction kinetic studies, detection or imaging techniques (such as positron emission tomography (PET) or single photon emission computed tomography (SPECT)), including drug or substrate tissue distribution assays, or in radiotherapy of patients.

[0112] The present disclosure also includes compounds I in which 1 to "n" hydrogens attached to a carbon atom are replaced by deuterium, where n is the number of hydrogens in the molecule. Such compounds exhibit increased metabolic resistance and can therefore be used to increase the half-life of any compound I when administered to a mammal. See, for example, Foster, "Deuterium Isotope Effects in Studies of Drug Metabolism," Trends Pharmacol. Sci. 5(12): 524-527 (1984). Such compounds are synthesized using methods well known in the art, such as by using a starting material in which one or more hydrogen atoms have been replaced by deuterium.

[0113] Deuterium-labeled or substituted therapeutic compounds of the present disclosure may have improved DMPK (drug metabolism and pharmacokinetics) properties, which are related to distribution, metabolism and excretion (ADME). Isotope-labeled compounds of the present disclosure and their prodrugs can generally be prepared by carrying out the processes disclosed in the following schemes or in the following examples and preparations, which are prepared by replacing non-isotope-labeled reagents with readily available isotope-labeled reagents. Further, the use of heavier isotopes, particularly deuterium (i.e. 2 Substitution by H or D) may provide certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements or improved therapeutic index. Isotopic labeling for diagnostic purposes is also contemplated.

[0114] The concentration of such isotopes, specifically deuterium, can be defined by isotopic enrichment factors. In the compounds of the present disclosure, any atom not specifically designated as a particular isotope is intended to represent any isotope of that atom. Unless otherwise indicated, when a position is specifically designated as "H" or "hydrogen", the position is understood to contain hydrogen in its natural abundance isotopic composition. Therefore, in the compounds of the present disclosure, any atom specifically designated as deuterium (D) is intended to represent deuterium greater than natural abundance.

[0115] As used herein, "pharmaceutically acceptable carriers" include excipients or agents that are not harmful to the compounds of the invention or their use, such as solvents, diluents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, etc. The use of such carriers and agents to prepare compositions of pharmaceutically active substances is well known in the art (see, for example, Remington's Pharmaceutical Sciences, Mace Publishing Co., Philadelphia, Pa. 17th edition (1985); and Modern Pharmaceutics, Marcel Dekker Company, 3rd edition, (GS Banker & C. T. Rhodes, Eds.).

[0116] The term "therapeutically effective amount" refers to an amount of a compound as described herein sufficient to achieve treatment as defined below when administered in one or more doses to a patient (particularly a human) in need of such treatment. The therapeutically effective amount will vary depending on the patient, the disease being treated, the patient's weight and / or age, the severity of the disease, or the mode of administration determined by a qualified prescriber or caregiver.

[0117] The terms "treatment" or "treating" refer to the administration of a compound described herein for the purpose of:

[0118] (i) delaying the onset of disease, i.e., preventing or delaying the development of clinical symptoms or markers of disease;

[0119] (ii) inhibiting the disease, i.e., slowing or preventing the development of clinical symptoms or markers thereof, or the spread of the disease;

[0120] and / or

[0121] (iii) ameliorating the disease, i.e. causing regression of clinical symptoms or markers of severity.

[0122] "Prevention" or "preventing" refers to the treatment of any disease or condition that results in the clinical symptoms of the disease or condition not developing. In some embodiments, the compound can be administered to a subject (including a human) who is at risk for a disease or condition or has a family history of the disease or condition.

[0123] "Subject" refers to an animal, such as a mammal (including a human), that has been or will be the subject of treatment, observation, or experiment. The methods described herein can be used for human treatment and / or veterinary applications. In some embodiments, the subject is a mammal. In one embodiment, the subject is a human. When the subject is a human, the subject may be referred to as a "patient."

[0124] The methods described herein can be applied to cell populations in vivo or in vitro. "In vivo" means within a living individual, such as in an animal or human body. In this context, the methods described herein can be used therapeutically for an individual. "Ex vivo" means outside a living individual. Examples of ex vivo cell populations include in vitro cell cultures and biological samples, including fluid or tissue samples obtained from an individual. Such samples can be obtained by methods well known in the art. Exemplary biological fluid samples include blood, cerebrospinal fluid, urine, and saliva. In this context, the compounds and compositions described herein can be used for a variety of purposes, including therapeutic and experimental purposes. For example, the compounds and compositions described herein can be used in vitro for a given indication, cell type, individual, and other parameters to determine the optimal schedule and / or dosage for the administration of the compounds disclosed herein. The information collected from this use can be used for experimental purposes or in the clinic for setting up a regimen for in vivo treatment. Other ex vivo uses for which the compounds and compositions described herein may be suitable are described below, or will become apparent to those skilled in the art. The selected compound can be further characterized as a safe dose or a tolerable dose for checking human or non-human subjects. Such properties can be checked using methods generally known to those skilled in the art.

[0125] In addition, the abbreviations used herein have the following corresponding meanings:

[0126]

[0127]

[0128] Forms of Compound I

[0129] As generally described above, the present disclosure provides crystalline forms of Compound 1 and co-crystals thereof. Other forms are also further discussed herein.

[0130] Crystalline 2-(2-(2-(2-(4-(Benzo[d]thiazol-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)ethan-1-ol (Compound 1 Form I) is characterized by an X-ray powder diffraction pattern including the following peaks: about 4.6, about 20.8, and about 23.7 °2θ±0.2 °2θ, as measured using a wavelength of The diffraction pattern includes additional peaks at about 9.2, about 16.3, about 18.6, and about 19.6° 2θ ± 0.2° 2θ. Compound 1 Form I is also characterized by being substantially as follows Figure 1 Its complete X-ray powder diffraction pattern is shown. Compound 1 Form I can be characterized by one or more, e.g., 1, 2, 3, 4, 5, or 6 of the following XRPD peaks:

[0131] peak 4.6 9.2 18.6 19.6 20.8 23.7

[0132] In some embodiments, Compound 1 Form I is characterized by a differential scanning calorimetry (DSC) curve comprising an endothermic peak at about 70°C ± 2°C. Compound 1 Form I is also characterized by being substantially as Figure 2 Its complete DSC curve is shown.

[0133] Crystalline 2-(2-(2-(2-(4-(Benzo[d]thiazol-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)ethan-1-ol (Compound 1 Form II) is characterized by an X-ray powder diffraction pattern including the following peaks: about 9.2, about 13.8, and about 16.1 °2θ±0.2 °2θ, as measured using a wavelength of The diffraction pattern includes additional peaks at about 6.9, about 11.5, and about 18.4° 2θ ± 0.2° 2θ. Compound I Form II is also characterized by being substantially as follows Figure 3 Its complete X-ray powder diffraction pattern is shown. In some embodiments, Compound 1 Form II exists as a wet material. Compound 1 Form II can be characterized by one or more, e.g., 1, 2, 3, 4, 5, or 6 of the following XRPD peaks:

[0134] peak 6.9 9.2 11.5 13.8 16.1 18.4

[0135] Crystalline 2-(2-(2-(2-(4-(Benzo[d]thiazol-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)ethan-1-ol (Compound 1 Form III) is characterized by an X-ray powder diffraction pattern including the following peaks: about 7.7, about 10.3, and about 15.3 °2θ±0.2 °2θ, as measured using a wavelength of The diffraction pattern includes additional peaks at about 5.1, about 12.8, and about 18.0° 2θ ± 0.2° 2θ. Compound I Form III is also characterized by being substantially as follows Figure 4 Its complete X-ray powder diffraction pattern is shown. In some embodiments, Compound 1 Form III exists as a wet material. Compound 1 Form III can be characterized by one or more, e.g., 1, 2, 3, 4, 5, or 6 of the following XRPD peaks:

[0136] peak 5.1 7.7 10.3 12.8 15.3 18.0

[0137] Crystalline 2-(2-(2-(2-(4-(Benzo[d]thiazol-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)ethan-1-ol (Compound 1 Form IV) is characterized by an X-ray powder diffraction pattern including the following peaks: about 5.4, about 22.4, and about 23.3 °2θ±0.2 °2θ, as measured using a wavelength of The diffraction pattern includes additional peaks at about 10.8, about 18.9, about 23.9, and about 26.8° 2θ ± 0.2° 2θ. Compound 1 Form IV is also characterized by being substantially as follows Figure 5 Its complete X-ray powder diffraction pattern is shown. Compound 1 Form IV can be characterized by one or more, e.g., 1, 2, 3, 4, 5, or 6 of the following XRPD peaks:

[0138] peak 5.4 10.8 18.9 22.4 23.3 23.9 26.8

[0139] In some embodiments, Compound 1 Form IV is characterized by a differential scanning calorimetry (DSC) curve comprising an endothermic peak at about 48°C ± 2°C and an endothermic peak at about 59°C ± 2°C. Compound 1 Form IV is also characterized by being substantially as Figure 6 Its complete DSC curve is shown.

[0140] Crystalline 2-(2-(2-(2-(4-(Benzo[d]thiazol-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)ethan-1-ol (Compound I Form V) is characterized by an X-ray powder diffraction pattern including the following peaks: about 4.6, about 9.2, and about 23.1 °2θ±0.2 °2θ, as measured using a wavelength of The diffraction pattern includes additional peaks at about 13.8 and about 18.6° 2θ ± 0.2° 2θ. Compound I Form V is also characterized by being substantially as follows Figure 7 Its complete X-ray powder diffraction pattern is shown. Compound 1 Form IV can be characterized by one or more, e.g., 1, 2, 3, 4, 5, or 6 of the following XRPD peaks:

[0141] peak 4.6 7.0 9.2 13.8 18.4 23.1

[0142] In some embodiments, Compound I Form V is characterized by a differential scanning calorimetry (DSC) curve comprising an endothermic peak at about 70°C ± 2°C. Compound I Form V is also characterized by being substantially as Figure 8 Its complete DSC curve is shown.

[0143] Some embodiments are directed to compositions comprising a form of Compound 1 as described herein without any other form of Compound 1. In some embodiments, the composition comprises greater than 95% of a form of Compound 1 as described herein relative to other forms of Compound 1. In some embodiments, the composition comprises greater than 97% of a form of Compound 1 as described herein relative to other forms of Compound 1. In some embodiments, the composition comprises greater than 99% of a form of Compound 1 as described herein relative to other forms of Compound 1.

[0144] Some embodiments are directed to compositions comprising other forms of Compound 1 comprising crystalline Compound 1 Form I as described herein. In some embodiments, the composition comprises greater than 95% crystalline Compound 1 Form I as described herein relative to other forms of Compound 1. In some embodiments, the composition comprises greater than 97% crystalline Compound 1 Form I as described herein relative to other forms of Compound 1. In some embodiments, the composition comprises greater than 99% crystalline Compound 1 Form I as described herein relative to other forms of Compound 1.

[0145] In some embodiments, the composition comprises crystalline Compound 1 Form I at a purity greater than 95%. In some embodiments, the composition comprises crystalline Compound 1 Form I at a purity greater than 97%. In some embodiments, the composition comprises crystalline Compound 1 Form I at a purity greater than 99%.

[0146] Some embodiments relate to methods of preparing a form of Compound I as described herein. In some embodiments, the method is as described in the Examples provided herein.

[0147] In some embodiments, the method for preparing Compound 1 Form I is selected from solid vapor diffusion, anti-solvent addition, liquid vapor diffusion, slow cooling, slurry conversion, temperature cycling and slow evaporation. Solid vapor diffusion can be carried out at room temperature with a solvent selected from acetone, THF, EtOH, H2O, EtOAc, dioxane, toluene, DCM and acetonitrile. Anti-solvent addition can be carried out by adding an anti-solvent selected from IPA, MTBE, CPME and heptane to a solution of Compound 1 in a solvent selected from THF, MeOH, DCM and EtOAc. Slow cooling can be carried out by cooling a solution of Compound 1 in a solvent or solvent system selected from MEK, dioxane: toluene (optionally at a ratio of about 1: 1 v / v), EtOAc: acetonitrile (optionally at a ratio of 1: 1 v / v), DCM: IPA (optionally at a ratio of about 4: 1 v / v) and THF: heptane (optionally at a ratio of about 4: 1 v / v). Slow evaporation can be performed by forming a solution of Compound 1 in a solvent or solvent system selected from THF, EtOAc, MEK, acetonitrile, and dioxane:H2O (optionally in a ratio of about 9:1 v / v).

[0148] In some embodiments, Compound 1 Form I is stable at 30°C / 65% RH (closed or open) and 40°C / 75% RH (closed or open) for at least 4 weeks. In some embodiments, Compound 1 Form I retains at least 99% purity after storage for at least 4 weeks, optionally at 30°C / 65% RH (closed or open) or 40°C / 75% RH (closed or open).

[0149] Drug composition and dosage

[0150] The form of Compound I as described herein can be administered in the form of a pharmaceutical composition. Therefore, also provided herein are pharmaceutical compositions containing one or more forms of Compound I as described herein and one or more pharmaceutically acceptable vehicles, the vehicle being selected from carriers, adjuvants and excipients. Suitable pharmaceutically acceptable vehicles may include, for example, inert solid diluents and fillers, diluents comprising sterile aqueous solutions and various organic solvents, penetration enhancers, solubilizers and adjuvants. Such compositions are prepared in a manner well known in the pharmaceutical field. See, for example, Remington's Pharmaceutical Sciences, Mace Publishing Co., Philadelphia, Pa., 17th edition (1985); and Modern Pharmaceutics, Marcel Dekker, 3rd edition, (GS Banker and CT Rhodes, ed.). Pharmaceutical compositions may be administered alone or in combination with other therapeutic agents.

[0151] The pharmaceutical composition can be administered in single dose or multiple doses. The pharmaceutical composition can be administered by various methods, including, for example, rectal, buccal, intranasal and transdermal routes. In certain embodiments, the pharmaceutical composition can be administered by intra-arterial injection, intravenous, intraperitoneal ("ip"), parenteral, intramuscular, subcutaneous, oral, topical or as an inhalant.

[0152] One mode of use is parenteral, such as by injection. The pharmaceutical compositions described herein may be incorporated into forms for administration by injection including, for example, aqueous or oil suspensions or emulsions with sesame oil, corn oil, cottonseed oil or peanut oil as well as elixirs, mannitol, glucose or sterile aqueous solutions and similar pharmaceutical vehicles.

[0153] Oral administration can be another route of administering one or more forms of Compound I described herein. Administration can be by, for example, capsules or enteric-coated tablets. In the preparation of pharmaceutical compositions comprising one or more compounds described herein, the active ingredient is typically diluted with an excipient and / or enclosed in a carrier, which can be in the form of a capsule, sachet, paper or other container. When an excipient is used as a diluent, it can be in the form of a solid, semisolid or liquid material that acts as an excipient, carrier, or medium for the active ingredient. Thus, the composition can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (in the form of solid or liquid media), ointments (containing, for example, up to 10% by weight of active ingredient), soft and hard gelatin capsules, sterile injectable solutions, and sterile packaged powders.

[0154] Some examples of suitable excipients include lactose, glucose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinyl pyrrolidone, cellulose, sterile water, syrup and methylcellulose. The formulation may additionally include: lubricants, such as talc, magnesium stearate and mineral oil; wetting agents; emulsifying and suspending agents; preservatives, such as methyl and propyl hydroxybenzoates; sweeteners; and flavoring agents.

[0155] Compositions comprising one or more forms of Compound I described herein can be formulated to provide rapid, sustained or delayed release of the active ingredient after administration to a subject using procedures known in the art. Controlled release drug delivery systems for oral administration include osmotic pump systems and dissolution systems, which include reservoirs or drug-polymer matrix formulations coated with polymers. Examples of controlled release systems are given below: U.S. Patent Nos. 3,845,770; 4,326,525; 4,902,514; and 5,616,345. Another formulation for the methods disclosed herein employs a transdermal delivery device ("patch"). Such transdermal patches can be used to provide continuous or intermittent infusion of the forms of Compound I described herein in controlled amounts. The construction and use of transdermal patches for delivering medicaments are well known in the art. See, for example, U.S. Patent Nos. 5,023,252, 4,992,445, and 5,001,139. Such patches may be constructed for continuous, pulsatile, or on demand delivery of pharmaceutical agents.

[0156] For preparing solid compositions, such as tablets, the principal active ingredient may be mixed with a pharmaceutical excipient to form a solid preformulation composition containing a homogenous mixture of a form of Compound I described herein. When referring to these preformulation compositions as homogenous, the active ingredient is dispersed evenly throughout the composition so that the composition can be readily subdivided into equally effective unit dosage forms such as tablets, pills, and capsules.

[0157] Tablets or pills in the form of Compound I as described herein can be coated or otherwise compounded to provide a dosage with the advantage of prolonged action or to protect the stomach from acidic conditions. For example, a tablet or pill can include an inner dosage component and an outer dosage component, the latter being in the form of a film on the former. The two components can be separated by an enteric layer, which is used to resist disintegration in the stomach and allows the inner component to be passed intact into the duodenum or delayed release. A variety of materials can be used for such enteric layers or coatings, including many polymeric acids and mixtures of polymeric acids with such materials as shellac, hexadecanol, and cellulose acetate.

[0158] Compositions for inhalation or insufflation may include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents or mixtures thereof and powders. Liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described above. In some embodiments, the composition is administered by oral or nasal respiratory route for local or systemic effect. In other embodiments, the composition in a pharmaceutically acceptable solvent may be atomized by the use of an inert gas. The atomized solution may be inhaled directly from an atomizing device, or the atomizing device may be connected to a mask tent or an intermittent positive pressure breathing machine. The solution, suspension or powder composition may be preferably administered orally or nasally from a device that delivers the formulation in an appropriate manner.

[0159] The forms of Compound I as described herein can be administered in a pharmaceutically effective amount. For oral administration, each dosage unit may contain 1 mg to 2 grams, 1 mg to 1 gram, or 1 mg to 500 mg of Compound I. In some embodiments, the dosage is 1 mg to 250 mg of Compound I. In some embodiments, the dosage range of Compound I is about 20 mg twice a day to about 50 mg twice a day. In some embodiments, the dosage is 2 mg, 4 mg, 6 mg, 8 mg, 10 mg, 12 mg, 14 mg, 16 mg, 18 mg, 20 mg, 25 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 75 mg, 100 mg, 200 mg, or 500 mg of Compound I. However, it should be understood that the amount of compound actually administered is usually determined by the physician according to relevant circumstances, including the condition to be treated, the route of administration selected, and the co-administered compound, as well as, if applicable, the age, weight, response of the individual patient, the severity of the patient's symptoms, etc.

[0160] The form of the compound I of the present application or its composition can be used to apply once, twice, three times or four times a day using any suitable mode mentioned above. In addition, the form of the compound I of the present application or its composition can be applied once or twice a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks or once every six weeks. In some embodiments, the form of the compound I of the present application or its composition can be applied once a day for 4 weeks, 8 weeks, 12 weeks, 16 weeks, 20 weeks, 24 weeks, 28 weeks, 32 weeks, 36 weeks, 40 weeks, 44 weeks, 48 ​​weeks, 52 weeks or longer as needed.

[0161] The active agent of the present application, such as the form of Compound I described herein, or its pharmaceutical composition, the specific dosage level for any particular subject will depend on a variety of factors, including the activity of the specific compound used, age, body weight, general health, sex, diet, administration time, route of administration and secretion rate, drug combination, and the severity of the specific disease of the subject undergoing therapy. For example, the dosage can be expressed as milligrams of the compound described herein per kilogram of the subject's body weight (mg / kg). A dosage between about 0.1 and 150 mg / kg may be appropriate. In some embodiments, about 0.1 and 100 mg / kg may be appropriate. In other embodiments, a dosage between 0.5 mg / kg and 60 mg / kg may be appropriate. Normalization according to the subject's body weight can be particularly useful when adjusting the dosage between subjects of widely different sizes, such as when using drugs in both children and adult humans or when converting the effective dosage of non-human subjects (such as dogs) to a dosage suitable for human subjects.

[0162] The daily dose can also be described as the total amount of the compound described herein administered per dose or per day. The daily dose of the compound described herein or its salt can be between about 1 mg and 4,000 mg, between about 2,000 mg / day to 4,000 mg / day, between about 1 mg / day to 2,000 mg / day, between about 1 mg / day to 1,000 mg / day, between about 10 mg / day to 500 mg / day, between about 20 mg / day to 500 mg / day, between about 50 mg / day to 300 mg / day, between about 75 mg / day to 200 mg / day, or between about 15 mg / day to 150 mg / day.

[0163] When administered nasally, the total daily dose for a human subject may be between 1 mg and 1,000 mg, between about 1,000-2,000 mg / day, between about 10-500 mg / day, between about 50-300 mg / day, between about 75-200 mg / day, or between about 100-150 mg / day. In various embodiments, the daily dose is about 10 mg, about 30 mg, about 50 mg, about 75 mg, about 100 mg, about 200 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, or about 1000 mg, or a range of values ​​therebetween.

[0164] The forms of Compound 1 described herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, can be administered once, twice, three times, or four times a day using any suitable means described above. Likewise, administration or treatment can continue for several days; for example, for a cycle of treatment, typically treatment will continue for at least 7 days, 14 days, or 28 days. Treatment cycles are well known and often alternate between the remaining period of about 1 day to 28 days, typically about 7 days or about 14 days, or between cycles. In other embodiments, treatment cycles can also be continuous. Administration or treatment can continue indefinitely.

[0165] In certain embodiments, the method comprises administering to the subject an initial daily dose of about 1 mg to 800 mg of a form of a compound described herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, and increasing the dose incrementally until clinical efficacy is achieved. The dose may be increased in increments of about 5 mg, 10 mg, 25 mg, 50 mg, or 100 mg. The dose may be increased daily, every other day, twice a week, or once a week.

[0166] The forms of Compound I described herein, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof, can be administered under fed conditions. The term "fed conditions" or variations thereof refers to the consumption or ingestion of food in solid or liquid form or calories in any suitable form prior to or simultaneously with the administration of the active ingredient. For example, the forms of Compound I described herein, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof, can be administered to a subject (e.g., a patient) within a few minutes or hours of consuming calories (e.g., a meal). In some embodiments, the forms of Compound I described herein, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof, can be administered to a subject (e.g., a patient) within 5-10 minutes, about 30 minutes, or about 60 minutes of consuming calories.

[0167] Treatments and uses

[0168] Fascin

[0169] Described herein is a method for regenerating spine synapses lost due to neurodegenerative diseases by targeting cytoskeletal proteins by administering to a subject a form of compound 1 as described herein. Unexpectedly, inhibition of the cytoskeletal protein fascin 1 (FSCN1) was observed to result in rapid upregulation of dendritic spines in vivo and in vitro. Dendritic spines contain filamentous actin (F-actin), which is a cytoskeletal polymer that confers cell structure and subcellular specialization. Without wishing to be bound by theory, it is believed that dendritic spines require the formation of highly branched assemblies of F-actin, and the formation of such assemblies can be prevented or significantly reduced by bundling into parallel arrays by fascin 1.

[0170] In some embodiments, a method of binding fascin is provided, the method comprising contacting fascin with an effective amount of Compound 1 described herein or a pharmaceutically acceptable salt thereof. In some embodiments, the method inhibits fascin. It is believed that forms of Compound 1 or a pharmaceutically acceptable salt thereof can promote dendritic spine formation by inhibiting fascin.

[0171] Fascin is an important actin cross-linker that does not have amino acid sequence homology with other actin binding proteins. Three forms of fascin are found in vertebrates: fascin 1, which is widely found in the nervous system and elsewhere; fascin 2, which is found in retinal photoreceptor cells; and fascin 3, which is found only in the testis. In some embodiments, fascin is human fascin 1. Fascin has a molecular weight of 55 kDa and can act as a monomeric entity and cross-link actin filaments into straight, tight and rigid bundles, thereby imparting mechanical rigidity to actin bundles. It is believed that fascin holds parallel actin filaments together to form filopodia with a diameter of approximately 60-200 nm.

[0172] During neuronal development, long bundles of f-actin are believed to push out the membranes of neurons to form structures such as axons, dendrites, filopodia, and lamellipodia. Fascin is considered to be involved in the cytoskeletal reorganization of newborn dendritic processes. Therefore, actin bundles of fascin are generally considered to be required for the formation and extension of axons and dendrites. It was surprisingly found that inhibiting the activity of fascin in the formation of actin bundles can promote the formation of dendritic spines, the protrusion of the plasma membrane of dendritic cells.

[0173] Neuronal diseases and conditions and treatment thereof

[0174] The unifying feature of neurodegenerative conditions with cognitive components is the loss of synapses that utilize glutamate amino acids as neurotransmitters ("glutamatergic synapses"), which are believed to be the most numerous types of synapses in humans and other mammals. Importantly, about 90% of glutamatergic synapses involve postsynaptic dendritic spines. Most of the synapses lost in neurodegenerative diseases are synapses where axons contact dendritic spines, so-called "axonal synapses." Under normal circumstances, changes in the density, shape, and protein composition of dendritic spines affect the strength of synaptic transmission and are the basis for several forms of synaptic changes (i.e., "plasticity") involved in learning and memory, cognitive flexibility, adaptability to injury and disease, and other processes. It is believed that these changes in axonal synapses are important for the memory encoding function of structures such as the hippocampus. Therefore, it is believed that the early and progressive loss of dendritic spines in the hippocampus and other regions is the driving force for memory loss and cognitive decline in Alzheimer's disease and other dementias. The development of novel methods to regenerate spine density may have important implications for the treatment of many neurodegenerative and developmental cognitive disorders.

[0175] Dendritic spines are specialized protrusions that are responsible for receiving synaptic inputs and providing important functions in the communication between neurons. The morphology of dendritic spines and their overall density are related to synaptic function and are strongly involved in memory and learning. Cellular changes in brain cells may contribute to the pathogenesis of neuronal diseases. For example, abnormal levels (such as reduction) of dendritic spine density in the brain may contribute to the onset of neuronal diseases. Therefore, it is believed that changes or disorders of dendritic spines can affect synaptic function and play an important role in various neurological and psychiatric disorders, such as autism, fragile X syndrome, Parkinson's disease (PD) and Alzheimer's disease (AD). For example, in AD, increasing evidence shows that defects begin with changes in hippocampal synaptic function caused by amyloid β (Αβ) protein before neuronal loss. Therefore, targeting initial synaptic loss rather than late stage disease intervention strategies can provide better prognosis for the treatment of AD. In addition, since most cognitive disorders can cause abnormal forms and functions of dendritic spines, it will be desirable to use small molecules to directly target them to change or alleviate these spine changes. For example, fragile X syndrome is characterized by too many immature spines.

[0176] Provided herein are methods that can be used to promote dendritic spine generation. In some embodiments, the method includes administering to a subject an effective amount of a form of Compound I described herein or a pharmaceutical composition thereof, as described herein including embodiments. It can be observed that dendritic spine generation is an increase in the average number of spines per neuron or an increase in the unit length of neurons, which increase can be referred to as an increase in dendritic spine density. Dendritic spine generation can be observed as an improvement in dendritic spine morphology. For example, it can be observed that the improvement in dendritic spine morphology is an increase in the average size of the spine head. It can be observed that dendritic spine generation is an improvement in dendritic spine size, spine plasticity, spine motility, spine density and / or synaptic function. It can be observed that dendritic spine generation is an increase in the local spatial average of membrane potential. It can be observed that dendritic spine generation is Ca 2+ An increase in the postsynaptic concentration (e.g., average volume) of. It can be observed that dendritic spine generation is an increase in the average ratio of mature to immature spines. In some embodiments, the form of Compound I described herein or its pharmaceutical composition increases dendritic spine density relative to a control. In some embodiments, compared to that observed at the beginning of treatment, the form of Compound I described herein or its pharmaceutical composition increases dendritic spine density. In some embodiments, the increase in dendritic spine density leads to a decrease in the symptoms of a neuronal disease or disorder in a subject or patient. In some embodiments, the increase in dendritic spine density is explained by anatomical observations. In some embodiments, an increase in dendritic spine density is observed in primary hippocampal neurons.

[0177] In some embodiments, the average dendritic spine density increases by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, 250%, 300%, 400%, 500%, 750% or 1000% or any range between any two of the numbers, including endpoints, relative to the time of starting treatment with a form of Compound I described herein or a pharmaceutical composition thereof. In some embodiments, the dendritic spine density increases by about 50% to about 500% relative to the time of starting treatment with a form of Compound I described herein or a pharmaceutical composition thereof. In some embodiments, the dendritic spine density increases by about 100% to about 300% relative to the time of starting treatment with a form of Compound I described herein or a pharmaceutical composition thereof. In some embodiments, dendritic spine density increases by about 200% to about 300% relative to the time of initiation of treatment with a form of Compound 1 described herein, or a pharmaceutical composition thereof. In some embodiments, the duration of treatment with a form of Compound 1 described herein, or a pharmaceutical composition thereof, is 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 1 day, 3 days, 5 days, 7 days, 14 days, 28 days, 90 days, 180 days, or 365 days.

[0178] In some embodiments, the method increases spine density by promoting the formation of new spines. In some embodiments, relative to a control (e.g., spine density in the absence of a compound), the method increases average spine density by at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, 100%, 125%, 150%, 175%, 200%, 250%, 300%, 400%, 500%, 750% or 1000% or any range between any two of the numbers, including endpoints. In some embodiments, relative to a control (e.g., spine density in the absence of a compound), the method increases average spine density by about 50% to about 500%. In some embodiments, relative to a control (e.g., spine density in the absence of a compound), the method increases spine density by about 100% to about 300%. In some embodiments, the method increases spine density by about 200% to about 300% relative to a control (eg, spine density in the absence of the compound).

[0179] In some embodiments, the method increases the spine density by increasing neuron length. In some embodiments, relative to the time of starting treatment with a form of Compound I as described herein or its pharmaceutical composition, the method increases the average neuron length by about 100nm, 300nm, 500nm, 700nm, 1 micron, 2 microns, 3 microns, 4 microns, 5 microns, 7 microns, 10 microns, 15 microns, 20 microns, 25 microns or any range between any two numbers in the numbers, including endpoints. In some embodiments, relative to the control (e.g., the length of neurons in the absence of the compound), the method increases the average neuron length by about 500nm to about 25 microns. In some embodiments, relative to the control (e.g., the length of neurons in the absence of the compound), the method increases the neuron length by about 10% to about 300%. In some embodiments, relative to the control (e.g., the length of neurons in the absence of the compound), the method increases the neuron length by about 200% to about 300%.

[0180] In some embodiments, the method increases the average number of spines per neuron relative to the time of initiation of treatment with a form of Compound I described herein, or a pharmaceutical composition thereof. In some embodiments, the average number of spines per unit length of neurons increases by at least about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500, 600, 700, 800, 900, or about 1000 or more, or any range between any two of the numbers, including endpoints. In some embodiments, the time is 1 hour, 2 hours, 4 hours, 8 hours, 1 day, 3 days, 5 days, 7 days, 14 days, 28 days, 90 days, 180 days, or 365 days.

[0181] In some embodiments, forms of Compound 1 described herein, or pharmaceutical compositions thereof, can be used to treat neuronal diseases and disorders. Neuronal diseases are diseases or conditions in which the function of the nervous system of a subject becomes impaired. Neuronal diseases or disorders can be neurological diseases or disorders. Neuronal diseases or disorders can be associated with neurodegenerative diseases or disorders.

[0182] In one aspect, a method for treating a neuronal disease in a patient in need is provided, the method comprising administering to the patient a therapeutically effective amount of a form of Compound I as described herein or a pharmaceutical composition thereof. In some embodiments, the neuronal disease is Alzheimer's disease. In some embodiments, the neuronal disease is Parkinson's disease. In some embodiments, the neuronal disease is Parkinson's dementia. In some embodiments, the neuronal disease is autism. In some embodiments, the neuronal disease is fragile X syndrome. In some embodiments, the disease or disorder is associated with the accumulation of amyloid plaques (e.g., characterized by it). In some embodiments, the neuronal disease is traumatic brain injury. In some embodiments, the patient with the neuronal disease suffers from traumatic brain injury before, during, or after the onset of the neuronal disease. In some embodiments, the neuronal disease comprises damaged neurons. Damaged neurons may include atrophy or other reduction of the effective function of neurons. For example, it is known that Alzheimer's disease manifests as damaged neurons, especially cortical neurons, such as hippocampal neurons and neurons near the hippocampus. The loss of synapses may be associated with the loss of dendritic spines and neurodegeneration.

[0183] In some embodiments, the neuronal disease is associated with abnormal dendritic spine morphology, spine size, spine plasticity, spine motility, spine density and / or abnormal synaptic function. In some embodiments, the neuronal disease is associated with abnormal (e.g., reduced) levels of dendritic spine density.

[0184] In some embodiments, the neuronal disease is Alzheimer's disease. In some embodiments, the neuronal disease is Parkinson's disease. In some embodiments, the neuronal disease is Parkinson's disease with dementia. In some embodiments, the neuronal disease is autism. In some embodiments, the neuronal disease is stroke. In some embodiments, the neuronal disease is post-traumatic stress disorder (PTSD). In some embodiments, the neuronal disease is traumatic brain disorder (TBD). In some embodiments, the neuronal disease is chronic traumatic encephalopathy (CTE). In some embodiments, the neuronal disease is schizophrenia. In some embodiments, the neuronal disease is dementia (e.g., common dementia). In some embodiments, the neuronal disease is attention deficit / hyperactivity disorder (ADHD). In some embodiments, the neuronal disease is amyotrophic lateral sclerosis (ALS). In some embodiments, the neuronal disease is frontotemporal lobar degeneration (FTLD) (e.g., FTLD-tau, FTLD-TDP or FTLD-FUS). In some embodiments, the neuronal disease is memory loss. In some embodiments, the neuronal disease comprises memory loss. In some embodiments, the neuronal disease is age-related memory loss. In some embodiments, the neuronal disease comprises age-related memory loss. In some embodiments, the neuronal disease is hypertensive encephalopathy. In some embodiments, the neuronal disease is chronic stress. In some embodiments, the neuronal disease comprises chronic stress. In some embodiments, the neuronal disease is FTLD-TDP type I. In some embodiments, the neuronal disease is FTLD-TDP type II. In some embodiments, the neuronal disease is FTLD-TDP type III. In some embodiments, the neuronal disease is FTLD-TDP type IV.

[0185] Examples of neuronal diseases that can be treated with a form of Compound 1 described herein, or a pharmaceutical composition thereof, or the methods described herein include Alexander's disease, Alper's disease, Alzheimer's disease, depression, perinatal asphyxia, Parkinson's disease dementia ("PD dementia"), amyotrophic lateral sclerosis, ataxia telangiectasia, Batten disease (also known as Spielmeyer-Vogt-Sjogren-Battenberg disease), spongiform encephalopathy (e.g., bovine spongiform encephalopathy (mad cow disease), kuru disease, Creutzfeldt-Jakob disease, fatal familial insomnia, Canavan disease, Cockayne syndrome, corticobasal degeneration, fragile X syndrome, frontotemporal dementia, Gerstmann-Straussler-Scheinker syndrome, syndrome), Huntington's disease, HIV-related dementia, Kennedy's disease, Krabbe's disease,Lewy body dementia, Machado-Joseph disease (spinocerebellar ataxia type 3), multiple sclerosis, multiple system atrophy, narcolepsy, neuroborreliosis, Parkinson's disease, Pelizaeus-Merzbacher disease, Pick's disease, primary lateral sclerosis, prion disease, Refsum's disease, Sandhoff's disease, Schilder's disease, subacute combined degeneration of the spinal cord secondary to pernicious anemia, schizophrenia, spinocerebellar ataxia (various types with different characteristics), spinal muscular atrophy, Steele-Richardson-Olszewski disease, Tabes dorsalis, drug-induced parkinsonism, progressive supranuclear palsy, corticobasal degeneration, multiple system atrophy, idiopathic Parkinson's disease, autosomal dominant Parkinson's disease, familial type 1 (PARK1), Parkinson's disease 3, autosomal dominant Lewy bodies (PARK3), Parkinson's disease 4, autosomal dominant Lewy bodies (PARK4), Parkinson's disease 5 (PARK5), Parkinson's disease 6, autosomal recessive early onset (PARK6), Parkinson's disease 2, autosomal recessive early onset (PARK2), Parkinson's disease 7, autosomal recessive early onset (PARK7), Parkinson's disease 8 (PARK8), Parkinson's disease 9 (PARK9), Parkinson's disease 10 (PARK10), Parkinson's disease 11 (PARK11), Parkinson's disease 12 (PARK12), Parkinson's disease 13 (PARK13), or mitochondrial Parkinson's disease. In some embodiments, the neuronal disease is Alzheimer's disease, Parkinson's disease, Parkinson's dementia, autism, stroke, post-traumatic stress disorder (PTSD), traumatic brain disorder (TBD), chronic traumatic encephalopathy (CTE), schizophrenia, dementia (e.g., common dementia), attention-deficit / hyperactivity disorder (ADHD), amyotrophic lateral sclerosis (ALS), frontotemporal lobar degeneration (FTLD) (e.g., FTLD-tau, FTLD-TDP, or FTLD-FUS), memory loss (e.g., age-related memory loss), hypertensive encephalopathy, or chronic stress. ,

[0186] In some embodiments, the neuronal disease is Alzheimer's disease (AD). Alzheimer's disease is characterized by symptoms of memory loss in the early stages of the disease. Apoε4 carriers are at greater risk of AD. It is generally believed that APOε4 is not as effective as other isotypes in clearing Aε, and therefore will be associated with greater amyloid load, tau phosphorylation, synaptic toxicity and reduced synaptic density. Having experienced traumatic brain injury (TBI) is another risk factor for AD, and studies have shown that those who experience TBI have a significantly increased risk of AD. Cognitive decline has been associated with the progressive loss of synapses. As the disease progresses, symptoms include confusion, long-term memory loss, paraphasia, vocabulary loss, aggressiveness, irritability and / or mood swings. In the later stages of the disease, there is a loss of physical function. Patients with Alzheimer's disease (AD) show many characteristic neuropathies, such as increased oxidative stress, mitochondrial dysfunction, synaptic dysfunction, destruction of calcium homeostasis, deposition of senile plaques and neurofibrillary tangles, and brain atrophy. Without wishing to be bound by any theory, it is believed that both the cause and effect of these neurological diseases are the accumulation of harmful forms of aggregated amyloid β (Αβ) peptides in the brain. AD-related conditions include: senile dementia of AD type (SDAT), frontotemporal dementia (FTD), vascular dementia, mild cognitive impairment (MCI), and age-associated memory impairment (AAMI). In some embodiments, a method for treating or preventing Alzheimer's disease is provided, the method comprising administering to a patient in need thereof a therapeutically effective amount of a form of Compound I described herein or a pharmaceutically acceptable salt thereof. In some embodiments, the patient is an Apoε2 or Apoε3 carrier. In some embodiments, the patient has suffered from TBI. In some embodiments, the patient is an Apoε4 carrier. In some embodiments, the patient is an Apoε4 carrier suffering from TBI.

[0187] In some embodiments, neuronal disease is fragile X syndrome (FXS). As known in the art, FXS is a genetic syndrome related to multiple disorders (e.g., autism and hereditary intellectual disability). The range of a series of values ​​that disability can present is mild to severe. Observe that in the task of performing the task that requires working memory, the male with FXS begins to gradually have more serious problems, and this usually starts after 40 years old. Observe that this is especially true for verbal working memory. In some embodiments, neuronal disease is autism. As known in the art, autism is a neurodevelopmental disorder. Without wishing to be bound by any theory, it is believed that autism affects the information processing in the brain by changing how nerves and synapses are connected and organized.

[0188] In further embodiments, compositions and methods are provided for alleviating, reducing or reversing a symptom of a neuronal disease or disorder.The symptom may be any symptom described herein.

[0189] The term "memory" and the like refer in the usual and customary sense to the process by which a subject encodes, stores and retrieves information. In the context of memory, the terms "encoding", "registering", etc. refer in the usual and customary sense to receiving, processing and combining information that affects the sensation caused by chemical or physical stimulation. In this context, the terms "storage", etc. refer in the usual and customary sense to the creation of a record of the encoded information. In this context, the terms "retrieval", "recall", etc. refer in the usual and customary sense to recalling the stored information. As known in the art, retrieval can be a response to a prompt. In some embodiments, memory loss refers to a reduction in the ability to encode, store or retrieve information. In some embodiments, memory can be recognition memory or recall memory. In this context, "recognition memory" refers to the recollection of previously encountered stimuli. As known in the art, stimuli can be, for example, words, scenes, sounds, smells, etc. A broader category of memory is "recall memory", which requires retrieval of previously learned information, such as a list of a series of actions, words or numbers previously encountered by the subject, etc. Methods for evaluating the level of memory encoding, storage and retrieval displayed by the subject are well known in the art, including methods disclosed herein. For example, in some embodiments, the method improves the memory of a subject in need, wherein the subject suffers from a neuronal disease. In some embodiments, the method improves the memory of the subject. In some embodiments, the method treats the neuron or cognitive impairment of the subject. In some embodiments, the method treats the neuron of the subject. In some embodiments, the method treats the cognitive impairment of the subject.

[0190] Further, for any aspect disclosed herein, in some embodiments, the subject suffers from brain injury. The type of brain injury comprises brain injury (i.e., destruction or degeneration of brain cells), traumatic brain injury (i.e., damage caused by external force to the brain), stroke (i.e., vascular events that temporarily or permanently damage the brain by hypoxia) and acquired brain injury (i.e., brain injury that does not exist at birth). In some embodiments, the method improves the memory of the subject. In some embodiments, the method improves the learning of the subject. In some embodiments, the method treats the neurons or cognitive impairment of the subject. In some embodiments, the method treats the neurons of the subject. In some embodiments, the method treats the cognitive impairment of the subject.

[0191] In some embodiments, a method for promoting dendritic spine generation in a patient in need is provided, the method comprising administering to the patient a form of compound I described herein or a pharmaceutically acceptable salt thereof. In some embodiments, a method for treating or preventing a neuronal disease or condition is provided, the method comprising administering to a patient in need thereof a therapeutically effective amount of a form of compound I described herein or a pharmaceutically acceptable salt thereof. In some embodiments, a form of compound I or a pharmaceutically acceptable salt thereof for treating a neuronal disease or condition is provided. In some embodiments, a form of compound I or a pharmaceutically acceptable salt thereof for preparing a drug for treating a neuronal disease or condition is provided. In some embodiments, the neuronal disease or condition is selected from Alzheimer's disease, Parkinson's disease, Parkinson's dementia, autism, fragile X syndrome, and traumatic brain injury. In some embodiments, the neuronal disease or condition is Alzheimer's disease. In some embodiments, a form of compound I described herein or a pharmaceutical composition thereof inhibits cross-linking of f-actin. In some embodiments, a form of compound I described herein or a pharmaceutical composition thereof is anti-metastatic.

[0192] Combination therapy

[0193] In one embodiment, the forms of Compound 1 disclosed herein can be used in combination with one or more additional therapeutic agents that are used and / or being developed to treat a neuronal disease or disorder.

[0194] When used to treat or prevent the above-mentioned diseases and conditions, the forms of Compound 1 described herein, or pharmaceutical compositions thereof, can be administered together with one or more other therapeutic agents, e.g., other therapeutic agents approved for the treatment or prevention of a particular disease or condition, more particularly agents that are considered to constitute the current standard of care. Where combination therapy is contemplated, the active agents may be administered simultaneously, separately, or sequentially in one or more pharmaceutical compositions.

[0195] The latest strategy for treating AD comprises the generation or aggregation state of the specific isotype of control A β peptide.Other strategy comprises the toxic form of tau that prevents, reduces or removes phosphorylated.Other strategy relates to small molecule targeted enzyme, and the enzyme produces A β peptide by processing amyloid precursor protein, plays a role in the abundance of A β peptide in the attempt to reduce the brain.In addition, the information about the effect of non-amyloid neuropathy (such as sporadic inheritance of specific mutations in tau disease or apolipoprotein E gene) is increasing, which stimulates the other strategy of resisting neurodegeneration.

[0196] The one or more additional therapeutic agents can be tacrine, donepezil, galantamine, rivastigmine, memantine, levodopa, carbidopa, lisuride, rasagiline, tolcapone, entacapone, clozapine, desipramine, citalopram, nortriptyline, paroxetine, atomoxetine, venlafaxine, amantadine, donepezil, rivastigmine, memantine, levodopa, carbidopa, lisuride, rasagiline, tolcapone, entacapone, clozapine, desipramine, citalopram, nortriptyline, paroxetine, atomoxetine, venlafaxine, amantadine, donepezil, rivastigmine, rivastigmine, bromocriptine, cabergoline, pergolide, pramipexole, ropinirole, rotigotine, benserazide, selegiline, omigapil, CEP-1347, isradipine, DOPA, lithium, riluzole, levetiracetam, ezogabine, pregabalin, rufmamide, felbamate, carbamazepine, valproate, sodium valproatevalproate, lamotrigine, phenytoin, oxcarbazepine, ethosuximide, gabapentin, tiagabine, topiramate, vigabatrin, phenobarbital, primidone, clonazepam, interferon beta-la, interferon beta-lb, mitoxantrone, natalizumab, fmgolimod, natalizumab, teriflunomide, dimethyl fumarate fumarate), glatiramer, ATOH1 gene therapy, ozanezumab, arimoclomol, tirasemtiv, dexpramipexole, pridopidine or galantamine; or phosphoglycerate kinase (PGK) as described in US2018 / 0147263. In some embodiments, the one or more additional therapeutic agents may be an acetylcholinesterase inhibitor (AChEI), such as acotiamide, alpha-pinene, ambenonium, demecarium, DFP (diisopropyl fluorophosphate), donepezil, edrophonium, galantamine, huperzine A, lactucorin, ladostinigil, neostigmine, physostigmine, pyridostigmine, diisopropyl fluorophosphate (dyflos), echothiophate, rivastigmine, rosmarinic acid (rosmarinic acid), succinate, ...In some embodiments, the one or more additional therapeutic agents may be amyloid-clearing antibodies, such as bapineuzumab, solanezumab, gantenerumab, crenezumab, ponezumab, BAN2401, or aducanumab.

[0197] The one or more additional therapeutic agents may be a sedative-hypnotic such as chloral hydrate, estazolam, flurazepam hydrochloride, pentobarbital, pentobarbital sodium, phenobarbital sodium, secobarbital sodium, temazepam, zaleplon or zolpidem tartrate; an anticonvulsant such as acetazolamide sodium, carbamazepine, clonazepam, clorazepate dipotassium, diazepam, divalproex sodium, ethosuximide, fosphenytoin sodium, or sedative. sodium, gabapentin, lamotrigine, magnesium sulfate, phenobarbital, phenobarbital sodium, phenytoin, phenytoin sodium, primidone, tiagabine hydrochloride, topiramate, valproate sodium, or valproic acid;Antidepressants, such as amitriptyline hydrochloride, amitriptyline pamoate, amoxapine, bupropion hydrochloride, citalopram hydrobromide, clomipramine hydrochloride, desipramine hydrochloride, doxepin hydrochloride, fluoxetine hydrochloride, imipramine hydrochloride, imipramine pamoate, mirtazapine, nefazodone hydrochloride, nortriptyline hydrochloride, paroxetine hydrochloride, phenelzine sulfate, sertraline hydrochloride hydrochloride, tranylcypromine sulfate, trimipramine maleate, or venlafaxine hydrochloride; antianxiety medicines such as alprazolam, buspirone hydrochloride, chlordiazepoxide, chlordiazepoxide hydrochloride, clorazepate dipotassium, diazepam, doxepin hydrochloride, hydroxyzine embonate, hydroxyzine hydrochloride, hydroxyzine pamoate, lorazepam, mephrobamate, midazolam hydrochloride, or oxazepam;Antipsychotics, such as chlorpromazine hydrochloride, clozapine, fluphenazine decanoate, fluephenazine enanthate, fluphenazine hydrochloride, haloperidol, haloperidol decanoate, haloperidol lactate, loxapine hydrochloride, loxapine succinate, mesoridazine besylate, molindone hydrochloride, olanzapine, perphenazine, pimozide, prochlorperazine, quetiapine fumarate fumarate, risperidone, thioridazine hydrochloride, thiothixene, thiothixene hydrochloride, or trifluoperazine hydrochloride; central nervous system stimulants such as caffeine, doxapram hydrochloride, methylphenidate hydrochloride, modafinil, pemoline, or phentermine hydrochloride;anti-Parkinson's disease agents, such as amantadine hydrochloride, benztropine mesylate, biperiden hydrochloride, biperiden lactate, bromocriptine mesylate, carbidopa-levodopa, entacapone, levodopa, pergolide mesylate, pramipexole dihydrochloride, ropinirole hydrochloride, selegiline hydrochloride, tolcapone, or trihexyphenidyl hydrochloride; or central nervous system agents, such as bupropion hydrochloride, donepezil hydrochloride, hydrochloride, droperidol, fluvoxamine maleate, lithium carbonate, lithium citrate, naratriptan hydrochloride, nicotine polacrilex, nicotine transdermal system, propofol, rizatriptan benzoate, sumatriptan succinate, tacrine hydrochloride, or zolmitriptan; cholinergics (e.g., parasympathomimetics) such as bethanechol chloride, edrophonium chloride, neostigmine bromide, neostigmine methylsulfate, methylsulfate), physostigminesalicylate or pyridostigmine bromide;Anticholinergics (such as atropine sulfate, dicyclomine hydrochloride, glycopyrrolate, hyoscyamine, hyoscyamine sulfate, propantheline bromide, scopolamine, scopolamine butylbromide, or scopolamine hydrobromide); Adrenergics (sympathomimetics) (such as dobutamine hydrochloride, dopamine hydrochloride, metaraminol bitartrate, norepinephrine bitartrate, and sedatives) bitartrate, phenylephrine hydrochloride, pseudoephedrine hydrochloride, or pseudoephedrine sulfate; adrenergic blockers (sympatholytics), such as dihydroergotamine mesylate, ergotamine tartrate, methysergide maleate, or propranolol hydrochloride; skeletal muscle relaxants, such as baclofen, carisoprodol, chlorzoxazone, cyclobenzaprine hydrochloride, dantrolene sodium, methocarbamol, or tizanidine hydrochloride;neuromuscular blockers such as atracurium besylate, cisatracurium besylate, doxacurium chloride, mivacurium chloride, pancuronium bromide, pipecuronium bromide, rapacuronium bromide, rocuronium bromide, succinylcholine chloride, tubocurarine chloride, or vecuronium bromide;or corticosteroids such as betamethasone, betamethasone acetate or betamethasone sodium phosphate, betamethasone sodium phosphate, cortisone acetate, dexamethasone, dexamethasone acetate, dexamethasone sodium phosphate, fludrocortisone acetate, hydrocortisone, hydrocortisone acetate, hydrocortisone cypionate, hydrocortisone sodium phosphate, hydrocortisone sodium succinate, methylprednisolone, methylprednisolone acetate acetate, methylprednisolone sodium succinate, prednisolone, prednisolone acetate, prednisolone sodium phosphate, prednisolone tebutate, prednisone, triamcinolone, triamcinoloneacetonide, or triamcinolone diacetate.

[0198] Reagent test kit

[0199] Also provided herein are kits comprising a form of Compound 1 described herein or a pharmaceutical composition thereof, an optional second active agent, and suitable packaging. In one embodiment, the kit further comprises instructions for use. In one aspect, the kit comprises a form of Compound 1 described herein or a pharmaceutical composition thereof, and a label and / or instructions for using the pharmaceutical composition in a therapeutic indication, including a disease or condition described herein.

[0200] Also provided herein are articles of manufacture comprising a form of Compound 1 described herein, or a pharmaceutical composition thereof, in a suitable container. The container can be a vial, a jar, an ampoule, a preloaded syringe, a nebulizer, an aerosol dispensing device, a dropper, or an intravenous bag.

[0201] synthesis

[0202] In some embodiments, the present disclosure provides methods for synthesizing Compound 1.

[0203] The disclosed method and its conventional modification can be utilized to carry out the method of the present disclosure, and the modification will be apparent in view of the content disclosed herein and methods known in the art. In addition to the teachings herein, conventional and well-known synthetic methods can be used. The synthesis of typical compounds described herein, such as compound I, compound A, etc., or other formulas or compounds disclosed herein can be completed as described in the following examples. If available, reagents can be purchased commercially from, for example, SigmaAldrich or other chemical suppliers.

[0204] Typical embodiments of compounds according to the invention can be synthesized using the general reaction schemes described below.A given agent may be defined as a general class or category (eg, function or structure), which should be interpreted as including any agent matching the given descriptor.

[0205] The compounds disclosed herein can be prepared from readily available starting materials using, for example, the following general methods and procedures. It should be understood that, unless otherwise stated, other process conditions may also be used given typical or preferred process conditions (i.e., reaction temperature, time, molar ratio of reactants, solvent, pressure, etc.). Optimum reaction conditions may vary with the specific reactants or solvents used, but such conditions may be determined by conventional optimization procedures by those skilled in the art.

[0206] In addition, it will be apparent to those skilled in the art that conventional protecting groups may be those described herein. Suitable protecting groups for various functional groups and suitable conditions for protecting and deprotecting specific functional groups are known in the art. For example, many protecting groups are described in TW Greene and GM Wuts (1999) Protective Groups in Organic Synthesis, 3rd edition, Wiley, New York, and references cited therein.

[0207] The starting materials for the following reactions are generally known compounds or can be prepared by known methods or obvious modifications thereof. For example, many starting materials are available from suppliers such as Aldrich Chemical Co. (Milwaukee, Wisconsin, USA), Bachen (Torrance, California, USA), Emka-Chemce or Sigma (St. Louis, Missouri, USA). Other compounds can be prepared by procedures described in the following standard reference texts, or obvious modifications thereof: Fieser and Fieser's Reagents for Organic Synthesis, Vols. 1-15 (John Wiley and Sons, 1991), Rodd's Chemistry of Carbon Compounds, Vols. 1-5 and Supplementals (Elsevier Science Publishers, 1989), Organic Reactions, Vols. 1-40 (John Wiley and Sons, 1991), March's Advanced Organic Chemistry (John Wiley and Sons, 5th ed., 2001), and Larock's Comprehensive Organic Transformations.

[0208] (VCH Publishers Inc., 1989).

[0209] The term "inert organic solvent" or "inert solvent" refers to a solvent that is inert under the reaction conditions described in connection therewith (including, for example, benzene, toluene, acetonitrile, tetrahydrofuran ("THF"), dimethylformamide ("DMF"), chloroform, methylene chloride (or dichloromethane), ether, methanol, pyridine, acetic acid, etc.). Unless otherwise indicated, the reaction is carried out under an inert gas such as nitrogen or argon. The "solvent" does not have to be inert.

[0210] In each exemplary scheme, it may be advantageous to separate the reaction products from each other and / or from the starting materials. The desired product of each step or series of steps is separated and / or purified (hereinafter separated) to the desired homogeneity by techniques commonly used in the art. Typically, such separations involve multiphase extraction, crystallization from a solvent or solvent mixture, distillation, sublimation, or chromatography. Chromatography can involve a variety of methods, including, for example: reverse phase and normal phase; size exclusion; ion exchange; high, medium, and low pressure liquid chromatography methods and apparatus; small-scale analysis; simulated moving bed (SMB) and preparative thin or thick layer chromatography, as well as small-scale thin layer and flash chromatography techniques.

[0211] Another class of separation methods involves treating the mixture with a medium selected to bind or otherwise separate the desired product, unreacted starting materials, reaction byproducts, etc. Such media include adsorbents or absorbents such as activated carbon, molecular sieves, ion exchange media, etc. Alternatively, acids (in the case of basic materials) or bases (in the case of acidic materials), binding agents such as antibodies, binding proteins, selective chelating agents such as crown ethers, liquid / liquid ion extraction agents (LIX), etc.

[0212] The choice of an appropriate separation method depends on the nature of the materials involved. For example, boiling point and molecular weight in distillation and sublimation, the presence or absence of polar functional groups in chromatography, the stability of materials in acidic and basic media in multiphase extraction, etc. One skilled in the art will apply the technique most likely to achieve the desired separation.

[0213] Raw materials and reagents were purchased from Shanghai Nenghua Titan Technology Co., Ltd. and used directly without further purification. Oxygen and moisture sensitive reactions were carried out under a nitrogen atmosphere. The reaction was monitored by thin layer chromatography (TLC), and the thin layer chromatography plate was visualized by exposure to ultraviolet light. Elution was monitored at 254nm. Flash column chromatography was usually performed on silica gel (300-400 mesh). Unless otherwise stated, yields refer to separated chromatographic and spectrally homogeneous materials.

[0214] Scheme 1 represents an exemplary synthesis of Compound I and can be performed according to the embodiments described herein. It is contemplated that the exemplary synthesis shown in Scheme 1 may be particularly advantageous. For example, the synthesis avoids toxic reagents. The synthesis can also utilize milder reaction conditions and may require fewer purification steps (e.g., avoiding column chromatography). The synthesis can also provide higher yields. The specific reaction conditions and reagents used in Scheme 1 are discussed below.

[0215] Compound I can be synthesized according to the general scheme, Scheme 1:

[0216] Solution 1

[0217]

[0218] In some embodiments, a method for preparing Compound 1 or a pharmaceutically acceptable salt thereof is provided:

[0219]

[0220] Comprising contacting compound A with compound B under first reaction conditions comprising a halide to form compound I:

[0221]

[0222] In some embodiments of the method for preparing Compound I, the halide is an alkali halide. In some embodiments of the method for preparing Compound I, the alkali halide is an alkali iodide. In some embodiments of the method for preparing Compound I, the alkali iodide is potassium iodide. In some embodiments of the method for preparing Compound I, the first reaction condition further comprises an inorganic base. In some embodiments of the method for preparing Compound I, the inorganic base is potassium carbonate. In some embodiments of the method for preparing Compound I, the first reaction condition comprises a temperature of 65 to 120°C.

[0223] In some embodiments, the method of preparing Compound A and optionally Compound I comprises contacting Compound C with Compound D under second reaction conditions comprising a protic acid to form Compound A:

[0224]

[0225] In some embodiments of the methods of preparing Compound I, the protic acid is an organic acid. In some embodiments of the methods of preparing Compound I, the organic acid is acetic acid. In some embodiments of the methods of preparing Compound I, the protic acid is present in a molar ratio greater than stoichiometric relative to Compound C and Compound D. In some embodiments of the methods of preparing Compound I, the protic acid dissolves Compound C and Compound D. In some embodiments of the methods of preparing Compound I, acetic acid is a non-inert solvent.

[0226] In some embodiments, the method of preparing Compound B and optionally Compound I comprises contacting Compound E with p-toluenesulfonyl chloride under third reaction conditions comprising a silver salt to form Compound B:

[0227]

[0228] In some embodiments of the method for preparing Compound I, the silver salt is Ag2O. In some embodiments of the method for preparing Compound I, the third reaction condition further comprises an alkali metal iodide. In some embodiments of the method for preparing Compound I, the alkali metal iodide is potassium iodide.

[0229] Example

[0230] Example 1

[0231] Experimental procedures

[0232] Compound I was subjected to various crystallization conditions, including solid vapor diffusion, anti-solvent addition, liquid vapor diffusion, slow cooling, slurry conversion at various temperatures, temperature cycling, and slow evaporation. The crystalline form of Compound I was analyzed by X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA), while the metastable form was analyzed by XRPD. XRPD was performed using a Panalytical X'Pert3 Powder XRPD with a Si zero background stand. The 2θ position was calibrated according to a Panalytical Si reference standard disk. The instrument parameters used are listed in Table 1-1.

[0233] Table 1-1: Parameters used for XRPD analysis

[0234]

[0235] TGA data were collected using a TA Q500 and Q550 from TA Instruments. DSC was performed using a TA Q2000 from TA Instruments. The DSC was calibrated using an indium reference standard and the TGA was calibrated using a nickel reference standard. Table 1-2 lists the detailed parameters used in the TGA and DSC tests.

[0236] Table 1-2: Parameters used for TGA and DSC tests

[0237] parameter TGA DSC method Slow Rise Slow Rise Sample tray Platinum, Open Aluminum, curled temperature RT - desired temperature 25℃-desired temperature Heating speed 10℃ / min 10℃ / min Purge gas <![CDATA[N2]]> <![CDATA[N2]]>

[0238] PLM images were captured using a Zeiss Axio Scope.A1 microscope. PLM images of samples obtained in the single crystal growth experiment were taken at room temperature using a Shanghai Cewei PXS9-T stereo microscope.

[0239] DVS was measured by SMS (Surface Measurement System) DVS Intrinsic. Relative humidity at 25°C was calibrated based on the deliquescent points of LiCl, Mg(NO3)2 and KCl. The actual parameters of the DVS test are shown in Table 1-3.

[0240] Table 1-3: Parameters used for DVS testing

[0241]

[0242] An Agilent 1260 HPLC equipped with a VWD detector was used and the detailed chromatographic conditions for purity analysis are listed in Tables 1-4.

[0243] Table 1-4: HPLC conditions and parameters

[0244]

[0245] A total of five crystalline forms were observed for Compound I, including an anhydrous form (Form I), a hydrate / solvate (Form IV), and three metastable forms (Forms II, III, and V). Forms II and III only exist in a wet state, and no solids were isolated for thermal analysis. Form V solids were obtained, however, Form V became Form I when dried in a vacuum oven at about 50°C. A summary of the isolated forms and conditions is shown in Tables 1-5 and 1-6.

[0246] Table 1-5: Isolation conditions of compound I forms

[0247] method result Solid vapor diffusion Form I and Form V Antisolvent addition Forms I, II, II, IV and V, amorphous Liquid vapor diffusion Form I and Form V Slow cooling Forms I, II and V, amorphous Slurry conversion Form I and Form V Temperature Cycle Form I and Form V Slow evaporation Form I and Form III

[0248] Table 1-6: Compound I Form Separation and Brief Description

[0249] Crystalline form Weight loss (%) Endothermic (peak, ℃) describe Form I Ignore 70.3 Anhydrous Form II <![CDATA[Not applicable 1 > <![CDATA[Not applicable 1 > Metastable Form III <![CDATA[Not applicable 2 > <![CDATA[Not applicable 2 > Metastable Form IV 4.4 48.0,58.9 Solvate / hydrate Form V 6.6 45.7(transformation),70.2 Metastable

[0250] Example 2

[0251] Forms of Compound I

[0252] Compound 1 Form I

[0253] Compound 1 Form I is an anhydrous form of Compound 1, and it is believed to be the most thermodynamically stable polymorph of the compound. To date, Form I is the only stable anhydrate form of Compound 1 identified.

[0254] Compound 1 Form I was obtained by various methods. Figure 1 The XRPD results in show that Form I is crystalline. TGA / DSC curve ( Figure 2 and Figure 3 ) showed negligible weight loss and an endothermic peak at 70.3° C. Based on these characterization data, Form I was considered to be an anhydrate.

[0255] Compound 1 Form I was isolated using the methods provided herein.

[0256] Compound I Form II

[0257] Form II was obtained by various methods. XRPD results showed that Form II was crystalline. It only existed in the form of wet cake and became a mixture of Form II and C or a mixture of Form III and Form V during storage. In order to further study Form II, several attempts to re-prepare were made, and the results formed Form III, IV, V or amorphous forms, indicating that Form II was a metastable form. Therefore, DSC and TGA data of Form II were not obtained.

[0258] Compound I Form III

[0259] Form III was obtained by various methods. XRPD results showed that Form III was crystalline. The Form III solid obtained by slow evaporation in MeOH was rapidly converted to Form I after exposure to open air during XRPD analysis. In order to further study Form III, several attempts to re-prepare were made, resulting in the formation of Form I or amorphous form, indicating that Form III is a metastable form. Therefore, DSC and TGA data of Form III were not obtained.

[0260] Compound I Form IV

[0261] Form IV was obtained by adding an antisolvent of H2O to the MEK stock solution. XRPD results showed that Form IV was crystalline. The TGA / DSC curve showed a weight loss of 4.4% before 150°C and two endothermic peaks at 48.0 and 58.9°C. Based on these characterization data, Form IV is considered to be a hydrate or solvate.

[0262] Compound I Form V

[0263] Form V was obtained by various methods. XRPD results show that Form V is crystalline. The TGA / DSC curve shows that the weight loss is 6.6% before 150°C, and two thermal events occur at 45.7°C (transition, onset) and 70.2°C (melting, peak). The XRPD data also show that the Form V solid is converted to Form I after vacuum drying at about 50°C. Upon dehydration / desolvation at about 50°C, Compound I Form V is converted to Compound I Form I. These results indicate that Form V is a metastable hydrate / solvate that is converted to Form I after dehydration / desolvation.

[0264] Amorphous Compound I

[0265] Amorphous Compound 1 was observed under the conditions described herein.

[0266] Example 3

[0267] Compound I Methods

[0268] Solubility

[0269] The approximate solubility of Compound I in 20 single solvents was determined at room temperature. Approximately 2 mg of powder sample was added to a 3 mL glass vial. The corresponding solvent was added to each vial stepwise (50 μL → 100 μL → 300 μL → 1000 μL) until the powder solid was visually completely dissolved or a total volume of 1 mL of solvent was added. The approximate solubility values ​​were calculated based on sample mass, solvent volume, and experimental observations. The results summarized in Table 3-1 are used to guide the selection of solvents in subsequent polymorph screening experimental designs.

[0270] Table 3-1: Solubility of Compound I at room temperature; * indicates that the sample precipitated from the clear solution (S < 40 mg / mL) after being placed at room temperature for about 10 minutes, and additional solvent was added to completely dissolve the remaining solid

[0271] Solvents Solubility (mg / mL) Solvents Solubility (mg / mL) DMF S>44 EtOH 22>S>7.3* DCM S>38 acetone 18>S>6* THF S>36 Toluene 18>S>6* 1,4-Dioxane S>32 MIBK 8.3>S>2.5* MeOH 42>S>21* IPA 5.7>S>1.7* EtOAc 40>S>20* CPME S<2.6* 2-MeTHF 40>S>20* Heptane S<2.2* MEK 36>S>18* MTBE S<2* DMSO 36>S>18* <![CDATA[H2O]]> S<1.8* Acetonitrile 24>S>8* IPA S<1.9*

[0272] Solid vapor diffusion

[0273] Solid vapor diffusion experiments were performed using 9 different solvents. Approximately 15 mg of the starting material was weighed into a 4 mL vial, which was then placed into a 20 mL vial containing 3 mL of a volatile solvent. The 20 mL vial was then sealed with a lid and kept at room temperature for 7 days to allow the solvent vapor to interact with the sample. If the solid was completely dissolved in the solvent, a slow evaporation was performed at room temperature. The obtained solid was analyzed by XRPD. The results summarized in Table 3-2 indicate that Form I and Form V were observed.

[0274] Table 3-2: Summary of solid vapor diffusion experiments of Compound I; * indicates that the powder solid is completely dissolved in the solvent, so the solid sample is obtained by slow evaporation.

[0275] Solvents Solid form acetone Form I* THF Form I* EtOH Form I + Form V <![CDATA[H2O]]> Form I EtOAc Form I Dioxane Form I* Toluene Form I DCM Form I* Acetonitrile Form I

[0276] Antisolvent addition

[0277] Antisolvent addition experiments were performed under 12 conditions. Approximately 20 mg of the starting material was dissolved in 0.5 or 1.0 mL of solvent to obtain a clear solution. The solution was then filtered and the antisolvent was added at a rate of 0.2 mL per step for magnetic stirring. The antisolvent was added until no more solid precipitated or the total volume of the solvent reached 5.0 mL. If the solution remained clear, the sample was stirred at 5 ° C to induce precipitation. If precipitation did not occur at 5 ° C, slow evaporation was performed at room temperature to obtain a solid. The solid precipitate was separated by centrifugation for subsequent XRPD analysis. The results in Table 3-3 show that Forms I, II, III, IV and V as well as an amorphous form were observed.

[0278] Table 3-3: Summary of antisolvent addition experiments for Compound I; * indicates solid obtained by slow evaporation at room temperature

[0279]

[0280] 1: Leave sample in solvent mixture and reanalyze on day 3; 2: Prepare sample on 2nd attempt and analyze immediately; 3: Prepare sample on 3rd attempt and analyze immediately; 4: Leave sample in solvent mixture and reanalyze on day 2 of 2nd attempt

[0281] Liquid vapor diffusion

[0282] Liquid vapor diffusion experiments were performed under 6 conditions. Briefly, about 20 mg of the starting material was dissolved in 1 mL of solvent and filtered into a 5 mL vial using a PTFE syringe filter with a pore size of 0.45 μm. The filtrate was then placed in a 20 mL vial containing 3 mL of a volatile solvent. The 20 mL vial was sealed with a lid and kept at room temperature to allow the organic solvent vapor to interact with the solution for at least 14 days. The precipitate was then separated for XRPD analysis. The results summarized in Tables 3-4 indicate that Form I and Form II were observed.

[0283] Table 3-4: Summary of liquid vapor diffusion experiments of compound I

[0284]

[0285]

[0286] Slow cooling

[0287] Slow cooling experiments were performed in 7 solvents or solvent combinations. Briefly, about 20 mg of the starting material was dissolved in 0.5 or 1 mL of solvent at 50 ° C and filtered into a new vial using a PTFE syringe filter with a pore size of 0.45 μm. The filtrate was slowly cooled from 50 ° C to 5 ° C at a rate of 0.1 ° C / min. The obtained solid was kept constant at 5 ° C before centrifugation and XRPD analysis. If the solution remains clear, slow evaporation is performed to obtain a solid for XRPD analysis. The results are summarized in Tables 3-5. The results show that Forms I, III, V and amorphous forms were observed.

[0288] Table 3-5: Summary of slow cooling experiments of Compound I; * indicates that the solution remained clear after cooling, so slow evaporation was performed

[0289] Solvent (v / v) Solid form MeOH <![CDATA[Form III* 1 > MEK Form I* Dioxane: toluene, 1:1 Form I* Ethyl acetate:acetonitrile, 1:1 Form I* DCM:IPA,4:1 Form I + Form V* THF:heptane, 4:1 Form I* DMSO:CPME, 4:1 Gel, amorphous*

[0290] 1: After drying, it becomes form I

[0291] Slurry conversion

[0292] Slurry conversion experiments were performed at room temperature or 60°C in 17 solvent systems. Approximately 20 mg of the starting material was suspended in 0.35 mL of solvent for 7 days at the target temperature. The remaining solid was separated for XRPD analysis. If the sample was completely dissolved, additional solid was added until a total of approximately 100 mg of powdered solid was added. The solid was then separated by centrifugation and analyzed with XRPD. The results summarized in Tables 3-6 indicate that Form I and Form V were observed.

[0293] Table 3-6: Summary of slurry conversion experiments of compound I

[0294]

[0295]

[0296] 1: Additional solid (≤100 mg) was added; 2: The solution remained clear after adding ~100 mg of solid, and after 7 days, the solution was kept at 5 °C to obtain solid for analysis

[0297] Temperature Cycle

[0298] The temperature cycling experiment was performed under 6 conditions. In brief, about 20 mg of the starting material was dissolved in 0.35 mL of solvent and stirred magnetically. The suspension was kept at 50 ° C for 2 hours, cooled to 5 ° C at a rate of 0.05 ° C / min, kept at 5 ° C for 2 hours, then raised to 50 ° C at a rate of 3 ° C / min, kept constant for 2 hours, and then cooled to 5 ° C at a rate of 0.05 ° C / min. The sample was stored at 5 ° C before centrifugation. The results are summarized in Tables 3-7. Form I and Form V were formed.

[0299] Table 3-7: Summary of temperature cycling experiments of compound I

[0300]

[0301]

[0302] *: After drying, it becomes Form I

[0303] Slow evaporation

[0304] Slow evaporation experiments were performed under 7 conditions. Approximately 20 mg of the starting material was dissolved in 0.5 or 1.0 mL of solvent and filtered using a PTFE syringe filter with a pore size of 0.45 μm. The filtrate was covered and stored at RT. The resulting solid was collected for XRPD analysis. The results are summarized in Tables 3-8. It was shown that Form I and Form III crystalline forms were observed.

[0305] Table 3-8: Summary of slow evaporation experiments of Compound I; * indicates that the solution remained clear after cooling, so slow evaporation was performed

[0306] Solvent (v / v) Solid form THF Form I* DCM <![CDATA[Form III + additional peak* 1 > MeOH <![CDATA[Form III* 1 > EtOAc Form I MEK Form I* Acetonitrile Form I <![CDATA[1,4 - Dioxane: H2O, 9:1]]> Form I*

[0307] 1: After drying, it is converted into form I

[0308] Example 4

[0309] Solid-state stability of compound I

[0310] The solid state stability of Compound I was evaluated for 4 weeks at 30°C / 65%RH (open petri dish and closed container) and 40°C / 75%RH (open petri dish and closed container). The detailed procedures for stability evaluation are listed as follows: 1) weigh approximately 10 mg of Compound I starting material and place it in a 3 mL glass vial; 2) for open petri dish conditions, use Cover the vial and poke 6 holes. For the airtight condition, seal the vial with a lid and keep the vial under the corresponding condition; for the 40°C / 75%RH condition, place the vial in a 40°C / 75%RH stabilization chamber; for the 30°C / 65%RH condition, place the vial in a closed atmosphere generated by a 30°C biochemical saturated potassium iodide aqueous solution; 4) Collect samples after 3 days, 1 week, 2 weeks and 4 weeks for XRPD, PLM and HPLC purity.

[0311] According to the XRPD results, no form change was observed after storage under all the above conditions. PLM characterization results showed that the compound I sample was still irregular flaky crystals with agglomeration. HPLC analysis showed that no significant decrease in HPLC purity was observed after 4 weeks at 30°C / 65% RH (open and closed) and 40°C / 75% RH (open and closed).

[0312] Solid-state stability evaluation of Compound 1 Form I starting material was performed under conditions of 30°C / 65% RH (closed and open) and 40°C / 75% RH (closed and open). X-ray powder diffraction (XRPD), polarizing microscopy (PLM), and HPLC purity were characterized at 4 sampling points (3 days, 1 week, 2 weeks, 4 weeks). No form changes and significant HPLC purity reduction of Compound 1 Form I were observed within 4 weeks under both conditions of 30°C / 65% RH and 40°C / 75% RH. The results of the solid-state stability evaluation are summarized in Table 4-1.

[0313] For single crystal structure determination, 83 single crystal growth experiments were performed by different methods including slow evaporation, liquid vapor diffusion, slow cooling, heating-cooling and solvent-thermal synthesis. Stability evaluation experiments were performed using a stability chamber at 40°C / 75% RH, while 30°C / 65% RH conditions were established in a 30°C biochemical incubator using a saturated potassium iodide aqueous solution.

[0314] Table 4-1: Summary of Solid State Stability Evaluation of Compound 1 Form I (HPLC Purity)

[0315]

[0316]

[0317] The Compound 1 starting material was a delivered sample. The starting material was characterized by XRPD, thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), PLM, and dynamic vapor sorption (DVS).

[0318] The TGA / DSC results of the compound I starting material showed that the weight loss was 0.45% before 150°C, and an obvious endotherm appeared at 68.2°C (starting temperature), which may be due to melting. The PLM results showed that the starting material had an irregular morphology and agglomeration. DVS diagram ( Fig.12 ) showed a water absorption of 0.64% at 25°C / 80% RH and 21.43% at 25°C / 95% RH. No form change was observed after DVS testing, as Fig.13 The sample information and characterization results are summarized in Table 4-2.

[0319] Table 4-2: Summary of characterization of starting materials of Compound I

[0320] weight Weight loss (%, up to 150°C) Endothermic (℃, start) 1.08g 0.45 68.2

[0321] Example 5

[0322] Single crystal growth

[0323] A total of 83 single crystal growth experiments of compound I were performed by different methods including slow evaporation, liquid vapor diffusion, slow cooling, heating-cooling and solvent-thermal synthesis. However, only flake-like and overlapping crystal samples were obtained, which were too thin for SCXRD characterization. Typical experimental procedures and details are described.

[0324] Slow evaporation

[0325] A total of 40 slow evaporation experiments were performed, and some single crystal samples with flake-like morphology were obtained. Typical experimental steps: Weigh the compound I starting material into a 3 mL glass vial, and add the selected solvent to the vial to dissolve the solid (use a vortex oscillator or an ultrasonic instrument to accelerate the dissolution). Filter the solution into a 4 mL shell vial (44.6 mm × 14.65 mm) using a PTFE filter (0.45 μm) and a disposable syringe. In some experiments, a small amount of compound I starting material was added as a seed crystal. Subsequently, the shell vial was covered with a PE-Plug with a pinhole on it and placed at the corresponding temperature for slow evaporation. After a few days, the sample was observed by PLM. Detailed experimental information is listed in Table 5-1, Table 5-2, Table 5-3 and Table 5-4.

[0326] Table 5-1: Slow evaporation single crystal growth experiment of compound I

[0327]

[0328] a: Compound I; b: A small amount of Compound I was added as a seed crystal in this experiment; c: Experimental results were observed after slow evaporation for 4 days

[0329] Table 5-2: Slow evaporation single crystal growth experiment of compound I

[0330]

[0331]

[0332] a: Compound I; b: A small amount of Compound I was added as a seed crystal in this experiment; c: Experimental results were observed after slow evaporation for 3 days

[0333] Table 5-3: Slow evaporation single crystal growth experiment of compound I

[0334]

[0335] a: Compound I starting material; b: A small amount of Compound I was added as seed crystals in this batch of experiments; c: Experimental results were observed after slow evaporation for 1 day; d: Experimental results were observed after slow evaporation for 4 days

[0336] Table 5-4: Slow evaporation single crystal growth experiment of compound I

[0337]

[0338] a: Compound I starting material; b: No seed crystals were added in this experiment; c: Experimental results were observed after slow evaporation for 1 day

[0339] Slow evaporation

[0340] A total of 22 liquid vapor diffusion experiments were carried out, and some single crystal samples with flake-like morphology were obtained.

[0341] Typical experimental steps: Weigh the compound I starting material into a 3 mL glass vial, add the selected solvent to the vial to dissolve the solid (a vortex oscillator or ultrasonic instrument can be used to accelerate dissolution). Filter the solution into a 4 mL shell vial (44.6 mm × 14.65 mm) using a PTFE filter (0.45 μm) and a disposable syringe. In some experiments, a very small amount of compound I starting material was added as a seed to induce crystallization. Subsequently, the shell vial was covered with a PE-Plug with a pinhole on it. The 4 ml shell vial was then placed in a 20 ml glass vial containing 3.0 ml anti-solvent, sealed and stored at the corresponding temperature for liquid vapor diffusion. After a few days, the sample was observed by PLM. Detailed experimental information is listed in Table 5-5, Table 5-6 and Table 5-7.

[0342] Table 5-5: Liquid vapor diffusion single crystal growth experiment of compound I

[0343]

[0344] a: Compound I starting material; b: A small amount of Compound I was added as a seed crystal in this group of experiments; c: Experimental results observed after 3 days of liquid vapor diffusion

[0345] Table 5-6: Liquid vapor diffusion single crystal growth experiment of compound I

[0346]

[0347] a: Compound I starting material; b: A small amount of Compound I was added as a seed crystal in this batch of experiments; c: Experimental results observed after 4 days of liquid vapor diffusion

[0348] Table 5-7: Liquid vapor diffusion single crystal growth experiment of compound I

[0349]

[0350] a: Compound I starting material; b: No seed crystals were added in this batch of experiments; c: EtOAc was saturated with H2O; d: Experimental results observed after 2 days of liquid vapor diffusion

[0351] Slow cooling

[0352] A total of 6 slow cooling experiments were carried out, and some single crystal samples with flake morphology were obtained.

[0353] Typical experimental procedure: The compound I starting material is weighed into a 3 ml glass vial and 0.5 mL of the selected solvent is added. After accelerating the dissolution process by a vortex oscillator or an ultrasonic device, the suspension is kept in a 50 ° C oven for about 0.5 hours. The hot solution is then filtered into another 3 ml glass bottle with a PTFE filter (0.45 μm) and a 2.0 ml disposable syringe (PTFE filter, disposable syringe and 3 ml glass bottle are preheated at 50 ° C), and a small amount of starting material is added to the vial as a seed. The vial is then sealed and placed in a biochemical incubator for slow cooling (cooling program: 50 ° C → 5 ° C, 0.01 ° C / min).

[0354] Table 5-8: Slow cooling single crystal growth experiment of compound I

[0355]

[0356] a: Compound I starting material; b: Cooling procedure: 50°C → 5°C, 0.01°C / min; c: Observation of experimental results after slow cooling for 4 days (cooling is completed)

[0357] Heating-Cooling

[0358] A total of 9 heating-cooling experiments were carried out, and some single crystal samples with flake morphology were obtained.

[0359] Typical experimental procedure: The compound I starting material is weighed into a 3 ml glass vial and 0.5 mL of the selected solvent is added. After accelerating the dissolution process by a vortex oscillator or an ultrasonic device, the suspension is kept in a 50 ° C oven for about 0.5 hours. The hot solution is then filtered into another 3 ml glass bottle with a PTFE filter (0.45 μm) and a 2.0 ml disposable syringe (PTFE filter, disposable syringe and 3 ml glass bottle are preheated at 50 ° C). A small amount of starting material is added to the vial as a seed crystal. The vial is then sealed and placed in a biochemical incubator for heating-cooling (heating-cooling program: 50 ° C → 5 ° C, 0.05 ° C / min, 5 cycles).

[0360] Table 5-9: Heating-cooling single crystal growth experiment of compound I

[0361]

[0362] a: Compound I starting material; b: Cooling procedure: 50°C → 5°C, 0.05°C / min, 5 cycles; c: Observation of experimental results after slow cooling for 4 days (cooling is completed)

[0363] Solvent-heat

[0364] A total of 6 solvent-thermal experiments were performed, and some single crystal samples with needle-like morphology were obtained. Typical experimental procedure: Compound I starting material was weighed into a 3 ml glass vial, and 0.4 mL of the selected solvent was added. Subsequently, the vial was placed in a hydrothermal reactor, sealed and placed in an oven for solvent-thermal experiments (temperature program: 25 °C → 80 °C → 25 °C).

[0365] Table 5-10 Solvent-heat single crystal growth experiment of compound I

[0366]

[0367]

[0368] a: Compound I starting material; b: Temperature program: 25°C → 80°C → 25°C; c: Observation of experimental results after 3 days of reaction (cooling ended)

[0369] Example 6

[0370] synthesis

[0371] Compound E is tosylated to form compound B

[0372]

[0373] Compound B

[0374] Compound E (99.8 g) was dissolved in anhydrous DCM (2.0 L) and stirred at room temperature. After 5 minutes, potassium iodide (18.1 g), Ag2O (179.8 g) and p-toluenesulfonyl chloride (108.5 g) were added to the solution in sequence. The reaction mixture was stirred vigorously overnight under N2. After diatomaceous earth filtration to remove solids, the filtrate was concentrated and purified by column chromatography (PE: EA = 1: 1 to PE: EA = 1: 2 to DCM: MeOH = 25: 1) to obtain a colorless oily compound B (110.0 g, 61.4% yield). 1 H NMR (400MHz, CDCl3) δ7.75 (d, J = 8.0 Hz, 2H), 7.34 (d, J = 8.0 Hz, 2H), 4.18-4.15 (m, 2H), 3.72-3.59 (m, 14H).

[0375] Compound C and compound D are condensed to form compound A

[0376]

[0377] Compound A

[0378] In a 3-L flask, compound C (160.3g) and compound D (172.0g) were dissolved in AcOH (1.5L). The mixture was refluxed at 105°C for 3.5 hours. After cooling to room temperature, the solution was poured into ice water and then filtered. The filtrate was concentrated and purified by recrystallization using EtOH (~1.4L) to produce dark gray solid compound A (105.7g, 36.3% yield). Recrystallization procedure: The crude product was dissolved in 1.4L EtOH to form a solution at 80°C. The solution was refluxed for a few minutes and then slowly cooled to room temperature. The solid was collected and washed with EtOH. The solid was vacuum dried at 50°C for about 8 hours. 1 H NMR (400MHz, CD3OD) δ7.95-7.91(m,4H),7.51-7.47(m,1H),7.40-7.36(m,1H),6.93-6.91(m,2H).

[0379] Nucleophilic addition of compound B to compound A forms compound I

[0380]

[0381] Compound B

[0382] Compound I is synthesized by the condensation of compound A and compound B. First, the condensation of compound A and compound B was attempted on a small scale. Compound B (5.0 g), compound A (3.3 g), K2CO3 (4.0 g), KI (0.2 g) and dry DMF (60 mL) were loaded into a flask. The resulting mixture was heated at 80 ° C overnight under N2. After cooling to room temperature, the product was extracted into EA by washing the aqueous layer with EA. The EA solvent was removed under reduced pressure, and the residue was purified by column chromatography (PE: EA = 3: 1) to obtain a white powder (4.3 g, 74.0% yield). Then, the preparation scale of compound I was expanded to hundreds of grams. Compound B (105.0 g), compound A (68.5 g), K2CO3 (83.4 g), KI (5.0 g) and dry DMF (1.1 L) were loaded into a flask. The resulting mixture was heated at 80 ° C overnight under N2. After cooling to room temperature, the product was extracted into EA by washing the aqueous layer (about 5 L H2O) with EA (about 7 L). The solvent of EA was removed under reduced pressure and the residue was purified twice by column chromatography (PE:EA=3:1) to give a white powder (~110.0 g). Two batches of compound I were mixed into DCM to form a solution. The DCM solvent was evaporated under reduced pressure to give pure compound I (103.0 g, 80.8% yield). The structure of compound I is shown in Table 1. 1 H NMR and MS confirmation. MS (m / z): [M+H]+=404.1. 1HNMR (400MHz, CDCl3) δ8.18 (d, J = 8.0 Hz, 1H), 8.11 (d, J = 8.0 Hz, 2H), 7.82 (d, J = 4.0 Hz, 1H), 7.49-7.45 (m, 1H), 7.38-7.34 (m, 1H), 7.01-6.98 (m, 2H), 4.18-4.15 (m, 2H), 3.84-3.81 (m, 2H), 3.67-3.61 (m, 10H), 3.55-3.53 (m, 2H). The HPLC purity of Compound I was also tested (HPLC, 230nm and 254nm UV). When detected at 230nm, the HPLC purity of Compound I was determined to be 99.8 area % and 99.7 area % by HPLC at 254nm (Tables 6-1 and 6-2, respectively). The XRPD pattern of the prepared Compound 1 was consistent with Form 1.

[0383] Table 6-1: HPLC purity of compound I at 230 nm

[0384]

[0385]

[0386] Table 6-2: HPLC purity of compound I at 254 nm

[0387] Peak number RRT area% 1 1.00 99.68 2 1.06 0.09 3 1.49 0.24

[0388] The source of the material is consistent with Table 6-3

[0389] Table 6-3: Sources of Materials

[0390]

[0391] Example 7

[0392] In vitro spinogenesis using benzothiazole compounds

[0393] To demonstrate the efficacy of the compounds in promoting spine formation, the effects of the compounds described herein on synaptic sites and synapses of mouse cortical neurons were studied.

[0394] Primary mouse cortical neurons were treated with 5 μM test compounds at DIV 15. As a control, primary mouse cortical neurons were treated with vehicle only (10% DMSO, 90% phosphate buffered saline (PBS)). After 24 hours, DIV 16 neurons were fixed, immunostained with the presynaptic vesicle protein synaptophysin (P38), counterstained with the nuclear dye DAPI (4',6-diamidino-2-phenylindole) and counted. Immunolabeled neurons were imaged on a Leica confocal microscope. The number of P38 immunopositive puncta was analyzed using FIJI with the Squash plugin.

[0395] In similar experiments, primary mouse cortical neurons were treated with 1 μM test compounds at DIV 15 using the same DMSO / PBS buffer solution as above as a control vehicle. 24 h later, DIV 16 neurons were fixed, immunolabeled with synaptophysin, stained with DAPI, and counted as above.

[0396] Example 8

[0397] Computer simulation of fascin binding

[0398] To evaluate the ability of the compounds described herein to bind to fascin and thereby inhibit its ability to bundle actin fibrils, computer simulation studies were performed using test compounds and available crystal structures of human fascin 1. Binding sites were identified on the surface of each fascin crystal structure, and then test compounds were virtually docked in each pocket to determine favorable binding conformations. See International Publication No. WO 2019 / 028164 (February 7, 2019).

[0399] Analysis and Preparation of Fascin Crystal Structure

[0400] All available fascin crystal structures were downloaded from the PDB and prepared for structural analysis (see Sedeh, R. S. et al. J. Mol. Biol. 400, 589–604 (2010); Chen, L. et al. Nature 464, 1062–1066 (2010); Jansen, S. et al. J. Biol. Chem. 286, 30087–30096 (2011); Yang, S. et al. J. Biol. Chem. 288, 274–284 (2013)). The structures were analyzed by eye and by standard automated protocols embedded in MolSoft's ICM-Pro software. Hydrogen atoms were added to the structures, and consideration was given to the correct orientation of Asn and Gln side chains, ligand and protein charge, histidine orientation and protonation state, and any crystallographic quality markers, such as high b-factors or low occupancy.

[0401] Pocket recognition

[0402] MolSoft's ICM PocketFinder algorithm was used to identify potential ligand binding pockets and cavities in all available fascin crystal structures (see An, J., et al. Genome Inform. Int. Conf. Genome Inform. 15, 31-41 (2004); Kufareva, I., et al. Nucleic Acids Res. 40, D535-540 (2012)). First, the pocket in the active chain A of the crystal structure 3LLP was searched because this structure was found to have the highest resolution. Four “drug-like” pockets were identified as having properties suitable for binding small molecules.

[0403] ■Ligand docking and scoring

[0404] The head group and head+tail of the test compound were docked into each of the four pockets using MolSoft's ICM-Docking software, version 3.8-6a (Abagyan, R. & Totrov, MJ Mol. Biol. 235, 983–1002 (1994)). The docking score for each pocket was determined. The lower the docking score, the better the "compound-fascin binding pocket" interaction.

[0405] Pocket B, located at actin binding site 1, was considered to result in the lowest docking score. Binding pocket B was further investigated in other fascin crystal structures. The docked head group was used as an anchor point, and the tail group was then docked to generate the final energetically favorable compound pose.

[0406] The scope of the present invention is not limited by the specific embodiments disclosed in the examples, which are intended to illustrate several embodiments of the invention, nor by any functionally equivalent embodiments within the scope of the present disclosure. Indeed, various modifications to the present disclosure, in addition to those shown and described herein, will become apparent to those skilled in the art and are intended to fall within the scope of the appended claims. To this end, it should be noted that one or more hydrogen atoms or methyl groups may be omitted from drawn structures consistent with the recognized shorthand notation for such organic compounds, and their presence will be readily understood by those skilled in the art of organic chemistry.

Claims

1. Crystalline 2-(2-(2-(2-(4-(Benzo[d]thiazol-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)ethan-1-ol (Compound I Form II), characterized in that The X-ray powder diffraction pattern includes the following peaks (°2θ±0.2°2θ): about 9.2, about 13.8 and about 16.1, using a wavelength of The Cu-Kα radiation was measured on a diffractometer.

2. The Compound 1 Form II of claim 1, wherein the diffraction pattern further comprises peaks at about 6.9, about 11.5, and about 18.4 (° 2θ ± 0.2° 2θ).

3. Crystalline 2-(2-(2-(2-(4-(Benzo[d]thiazol-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)ethan-1-ol (Compound I Form III), characterized in that The X-ray powder diffraction pattern includes the following peaks (°2θ±0.2°2θ): about 7.7, about 10.3 and about 15.3, using a wavelength of The Cu-Kα radiation was measured on a diffractometer.

4. The Compound 1 Form III of claim 3, wherein the diffraction pattern further comprises peaks at about 5.1, about 12.8, and about 18.0 (° 2θ ± 0.2° 2θ).

5. Crystalline 2-(2-(2-(2-(4-(Benzo[d]thiazol-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)ethan-1-ol (Compound I Form IV), characterized in that The X-ray powder diffraction pattern includes the following peaks (°2θ±0.2°2θ): about 5.4, about 22.4 and about 23.3, using a wavelength of The Cu-Kα radiation was measured on a diffractometer.

6. The Compound 1 Form IV of claim 5, wherein the diffraction pattern further comprises peaks at about 10.8, about 18.9, about 23.9, and about 26.8 (° 2θ ± 0.2° 2θ).

7. Compound 1 Form IV according to claim 5, characterized in that The differential scanning calorimetry (DSC) curve includes an endotherm at about 48°C and an endotherm at about 59°C.

8. Crystalline 2-(2-(2-(2-(4-(Benzo[d]thiazol-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)ethan-1-ol (Compound I Form V), characterized in that The X-ray powder diffraction pattern includes the following peaks (°2θ±0.2°2θ): about 4.6, about 9.2, and about 23.1, using a wavelength of The Cu-Kα radiation was measured on a diffractometer.

9. Compound 1 Form V according to claim 8, wherein the diffraction pattern further comprises peaks at about 13.8 and about 18.6 (° 2θ ± 0.2° 2θ).

10. Compound I Form V according to claim 8, characterized in that The differential scanning calorimetry (DSC) curve included an endotherm at about 70°C.

11. A pharmaceutical composition comprising one or more crystalline forms as claimed in any one of the preceding claims and one or more pharmaceutically acceptable carriers.

12. Use of a form of Compound I as described in any one of claims 1 to 10 or a pharmaceutical composition as described in claim 14 for the preparation of a medicament for promoting spines in a patient.

13. The use according to claim 12, wherein the patient suffers from a neuronal disease.

14. Use according to claim 13, wherein the neuronal disease is selected from the group consisting of Alzheimer's disease, Parkinson's disease, Parkinson's dementia, autism, fragile X syndrome and traumatic brain injury.

15. Use according to claim 14, wherein the neuronal disease is Alzheimer's disease.

16. A method for preparing Compound I or a pharmaceutically acceptable salt thereof: Comprising contacting compound A with compound B under first reaction conditions comprising a halide to form compound I:

17. The method of claim 16, wherein the halide is an alkali metal halide.

18. The method of claim 17, wherein the alkali metal halide is an alkali metal iodide.

19. The method of claim 18, wherein the alkali metal iodide is potassium iodide.

20. The method of claim 16, wherein the first reaction condition further comprises an inorganic base.

21. The process of claim 20, wherein the inorganic base is potassium carbonate.

22. The method of claim 16, wherein the first reaction conditions comprise a temperature of 65 to 120°C.

23. The method of claim 16, further comprising contacting compound C with compound D to form compound A under second reaction conditions comprising a protic acid:

24. The method of claim 23, wherein the protic acid is an organic acid.

25. The method of claim 24, wherein the organic acid is acetic acid.

26. The method of claim 23, wherein the protic acid is present in a greater than stoichiometric molar ratio relative to Compound C and Compound D.

27. The method of claim 26, wherein the protic acid dissolves Compound C and Compound D.

28. The method of claim 23, further comprising contacting compound E with p-toluenesulfonyl chloride under third reaction conditions comprising a silver salt to form compound B:

29. The method of claim 28, wherein the silver salt is Ag2O.

30. The method of claim 28, wherein the third reaction condition further comprises an alkali metal iodide.

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