A2-73 crystalline polymorph compositions of matter and methods of use thereof

Abstract

Crystalline forms of tetrahydro-N, N-dimethyl-2, 2-diphenyl-3-furanmethanamine (A2 - 73), dosage forms containing them, and methods of using them in therapy are provided.SOLUTION: The present invention provides crystalline forms of tetrahydro-N, N-dimethyl-2, 2-diphenyl-3-furanmethanamine (A2 - 73) in free base or salified form. Also described are pharmaceutical formulations and dosage forms comprising the disclosed crystalline forms, and methods of using A2 - 73 in dosage forms for neuroprotection, including the treatment of neurodegenerative diseases and other diseases.SELECTED DRAWING: None

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No. 62 / 656,435, filed April 12, 2018, the entire disclosure of which is incorporated herein by reference.

[0002] The present invention relates generally to crystalline forms of tetrahydro-N,N-dimethyl-2,2-diphenyl-3-furanmethanamine (A2-73), dosage forms containing them, and methods of using them therapeutically. [Background technology]

[0003] Tetrahydro-N,N-dimethyl-2,2-diphenyl-3-furanmethanamine (ANAVEX2-73 or AV2-73) is a mixed muscarinic receptor ligand and Sig-1R agonist with affinity in the low micromolar range. A2-73 can treat neurodevelopmental disorders and has neuroprotective properties. Improved drug formulations that exhibit, for example, better bioavailability, greater stability, or enhanced delivery of pharmaceutically active compounds are consistently sought, and there is a continuing need for more fully characterized novel drug molecules. There is also a continuing need for methods to treat neurodegenerative diseases. Summary of the Invention

[0004] In one aspect, the present disclosure encompasses crystalline forms of tetrahydro-N,N-dimethyl-2,2-diphenyl-3-furanmethanamine (A2-73), wherein the crystalline form is a salt or a free base. The salt may be any pharmaceutically acceptable salt, such as hydrochloride, fumarate, sulfate, dihydrogen phosphate, benzoate, mesylate, edysilate, and oxalate. It is understood that in any of the pharmaceutical formulations, dosage forms, and methods disclosed herein, the crystalline A2-73 may be the free base disclosed herein or a salt disclosed herein, including any one or more of hydrochloride, fumarate, sulfate, dihydrogen phosphate, benzoate, mesylate, edysilate, and oxalate.

[0005] When A2-73 is a hydrochloride salt, the hydrochloride salt is characterized by the XRPD patterns shown in Figures 4, 6, 8, 9, 10, 11, 12, and 14. The hydrochloride salt characterized by the XRPD pattern shown in Figure 4 may be further characterized by the particle shape and size illustrated in Figures 2 and 3. The hydrochloride salt characterized by the XRPD pattern shown in Figure 6 may be further characterized by the particle shape and size illustrated in Figure 5. The hydrochloride salt characterized by the XRPD pattern shown in Figure 8 may be further characterized by the particle shape and size illustrated in Figure 7. The hydrochloride salt characterized by the XRPD pattern shown in Figure 14 may be further characterized by the particle shape and size illustrated in Figure 13.

[0006] The crystalline form of A2-73 can be the sulfate salt. The sulfate salt can be characterized by the XRPD patterns shown in Figures 18 and 19. The sulfate salt characterized by the XRPD pattern shown in Figure 18 can be further characterized by the particle shape illustrated in Figure 17.

[0007] The crystalline form of A2-73 can be the mesylate salt, which can be characterized by the XRPD pattern shown in Figure 20.

[0008] The crystalline form of A2-73 can be the oxalate salt, which can be characterized by the XRPD patterns shown in Figures 21, 22, and 23.

[0009] The crystalline form of A2-73 can be a dihydrogen phosphate salt. The dihydrogen phosphate salt can be characterized by the XRPD pattern shown in Figure 25. The dihydrogen phosphate salt characterized by the XRPD pattern shown in Figure 25 is further characterized by the particle shape depicted in Figure 24.

[0010] The crystalline form of A2-73 may be the edysilate salt, which may be characterized by the XRPD pattern shown in FIG.

[0011] The crystalline form of A2-73 may be the benzoate salt, which may be characterized by the XRPD pattern shown in Figure 27.

[0012] The crystalline form of A2-73 can be a fumarate salt. The fumarate salt can be characterized by the XRPD patterns shown in Figures 29, 30, 32, 33, and 34. The fumarate salt characterized by the XRPD pattern shown in Figure 29 can be further characterized by the particle shape shown in Figure 28, and the fumarate salt characterized by the XRPD pattern shown in Figure 32 can be further characterized by the particle shape shown in Figure 31.

[0013] The crystalline form of A2-73 can be the free base. The free base can be characterized by the XRPD pattern shown in Figure 16. The crystalline form characterized by the XRPD pattern shown in Figure 16 is further characterized by the particle shape depicted in Figure 15.

[0014] In another aspect, the disclosure encompasses a dosage form comprising a therapeutically effective amount of A2-73 in a crystalline form selected from the group consisting of A2-73 free base and A2-73 salts. The dosage form may contain from about 1 mg to about 50 g, from about 1 mg to about 500 mg, or from about 1 mg to about 100 mg of A2-73 free base or A2-73 salt.

[0015] The dosage form may be formulated for extended release of crystalline A2-73. In any dosage form, the crystalline A2-73 may be free base, and the dosage form may contain about 1 mg to about 500 mg of A2-73 free base. The dosage form may be a transdermal patch. The transdermal patch may contain about 40 mg to about 60 mg, about 80 mg to about 120 mg, or about 180 mg to about 220 mg of A2-73 free base. The dosage form may be an enteric-coated oral formulation, and the formulation may contain about 1 mg to about 50 mg of A2-73 free base.

[0016] In any dosage form, the crystalline A2-73 may be a pharmaceutically acceptable salt. The pharmaceutically acceptable salt may be selected from the group consisting of fumarate, sulfate, mesylate, dihydrogen phosphate, edisylate, benzoate, hydrochloride, and oxalate. In some embodiments, the A2-73 salt is a fumarate salt, and the dosage form may be a transdermal patch. The transdermal patch may contain about 1 mg to about 55 mg of A2-73 fumarate.

[0017] In some embodiments, the dosage form may be an enteric-coated oral formulation, which may contain about 10 mg to about 50 mg, about 20 mg to about 30 mg, or about 15 mg to about 25 mg of A2-73 fumarate.

[0018] In another embodiment, the disclosure encompasses a pharmaceutical formulation for the delivery of A2-73. , a therapeutically effective amount of a crystalline form of A2-73 selected from A2-73 free base and A2-73 salts.

[0019] The pharmaceutical formulation may further comprise one or more pharmaceutically acceptable non-medicinal excipients selected from chemical enhancers, humectants, pressure-sensitive adhesives, antioxidants, solubilizers, thickeners, plasticizers, adjuvants, carriers, excipients, vehicles, and any combination thereof. The one or more non-medicinal ingredients may be selected to prepare a formulation for oral, transdermal, parenteral, intraperitoneal, intravascular, subcutaneous, by inhalation spray, rectal, or pulmonary administration.

[0020] In any pharmaceutical formulation, crystalline A2-73 can be selected from the free base and any pharmaceutically acceptable salt. In one embodiment of the pharmaceutical formulation, crystalline A2-73 is a fumarate or hydrochloride salt. The pharmaceutical formulation can be, for example, an oral formulation containing about 1% to about 100% by weight of crystalline A2-73.

[0021] The pharmaceutical formulation may be prepared for extended delivery of crystalline A2-73 and may contain from about 1 mg to about 50 g of crystalline A2-73. The extended delivery formulation may be, for example, a subcutaneously injectable dosage formulation containing from about 0.5 g to about 3 g of crystalline A2-73.

[0022] The pharmaceutical formulation may be a transdermal patch. The patch may contain about 40 mg to about 60 mg, about 80 mg to about 120 mg, or about 180 mg to about 220 mg of A2-73 free base. The patch may also contain about 1 mg to about 55 mg of A2-73 fumarate.

[0023] The formulation may also be an oral formulation. The oral formulation may contain about 1 mg to about 50 mg of A2-73 free base, about 10 mg to about 50 mg, about 20 mg to about 30 mg, or about 15 mg to about 25 mg of A2-73 fumarate. The oral formulation may contain A2-73 hydrochloride.

[0024] The extended delivery formulation can be a subcutaneous dosage form containing about 0.1 to about 5 g of crystalline A2-73.

[0025] In yet another embodiment, the present disclosure encompasses a transdermal patch for extended delivery of A2-73. The patch may contain a therapeutically effective amount of a crystalline form of A2-73 selected from A2-73 free base and A2-73 salts. The transdermal patch may be, for example, a matrix patch. The patch may further contain one or more ingredients selected from chemical enhancers, humectants, pressure-sensitive adhesives, antioxidants, solubilizers, thickeners, plasticizers, and any combination thereof. The patch may be covered by a peripheral pressure-sensitive adhesive that extends beyond the patch in all directions.

[0026] A transdermal patch containing A2-73 free base can contain about 40 mg to about 60 mg, about 80 mg to about 120 mg, or about 180 mg to about 220 mg of A2-73 free base.

[0027] The transdermal patch may contain A2-73 fumarate. The patch containing A2-73 fumarate may contain about 1 mg to about 55 mg of A2-73 fumarate.

[0028] The surface area of ​​the transdermal patch in contact with the subject's skin is approximately 1 cm 2 ~about 20cm 2 , about 3cm 2 ~about 5cm 2 , or about 8 cm 2 ~about 10cm 2 The patch may be configured to provide extended release of A2-73 over a period ranging, for example, from about 1 day to about 7 days. Additionally, the patch may provide a dose of about 250-350 μg / cm. 2 The maximum transdermal flux of A2-73 from the matrix may be in the range of / h.

[0029] In other embodiments, the present disclosure encompasses oral formulations for extended delivery of A2-73. The oral formulation comprises a core comprising a therapeutically effective amount of a crystalline form of A2-73 selected from A2-73 free base and A2-73 salts, and an enteric coating surrounding the core.

[0030] The oral formulation may contain A2-73 free base, which may range from about 1 mg to about 50 mg of A2-73 free base. The oral formulation may also contain A2-73 fumarate in the core, which may range from about 10 mg to about 50 mg, about 20 mg to about 30 mg, or about 15 mg to about 25 mg, or A2-73 fumarate, which may range from about 35% to about 40% by weight of A2-73 free base or A2-73 fumarate. The oral formulation may also contain about 55% to about 70% by weight of hydroxypropylmethylcellulose acetate succinate, about 0.3% to about 0.9% by weight of magnesium stearate, and about 0.05% to about 0.5% by weight of colloidal silicon dioxide in the core. The hydroxypropyl methylcellulose acetate succinate may be soluble in aqueous solution having a pH of about 5.5 or higher, the second grade hydroxypropyl methylcellulose acetate succinate may be soluble in aqueous solution having a pH of about 6.8 or higher, and combinations thereof. The formulation may provide extended release of A2-73 over a period ranging from about 1 day to about 3 days, and may deliver about 15 to about 30 mg / day of A2-73 to a subject.

[0031] In one embodiment, the present disclosure encompasses a method of administering A2-73 to a subject in need thereof. The method includes administering A2-73 to the subject in a crystalline form of A2-73 selected from A2-73 free base and a pharmaceutically acceptable salt of A2-73. In various embodiments of the method, the crystalline A2-73 can be administered in a dosage form or pharmaceutical formulation containing crystalline A2-73 free base or a pharmaceutically acceptable salt of A2-73 as disclosed herein. The dosage form can be an immediate-release or extended-release dosage form as disclosed herein. In certain embodiments, the salt can be a fumarate salt or a hydrochloride salt. In other embodiments, the crystalline A2-73 can be administered to the subject for a period of about 30 days, about 60 days, about 120 days, or about 180 days.

[0032] In one embodiment, administration can include administration using an extended-release dosage form that can be administered transdermally using a transdermal patch. The transdermal patch can be replaced periodically, for example, daily, every other day, weekly, every 10 days to two weeks, or monthly, or more frequently. In one embodiment, the transdermal patch can maintain the A2-73 level in the subject's blood at a range of about 5 ng / ml to about 15 ng / ml, particularly about 10 ng / ml, over a period of time.

[0033] In another embodiment, administration can include administering using an enteric-coated oral dosage form containing crystalline A2-73. The enteric-coated oral dosage form can be administered daily or every other day and can deliver about 15 to about 30 mg of A2-73 per day. The enteric-coated oral dosage form containing crystalline A2-73 can provide administration of A2-73 for an extended period of time, which can be, for example, about 1 day, about 2 days (about 48 hours), 3 days (about 72 hours), about 4 days, about 5 days, about 6 days, about 7 days, or longer.

[0034] In another aspect, the disclosure includes a method of treating Alzheimer's disease in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a dosage form comprising a crystalline form of A2-73 selected from A2-73 free base and A2-73 salt.

[0035] In another aspect, the disclosure includes a method of treating progressive dementia in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a dosage form comprising a crystalline form of A2-73 selected from A2-73 free base and A2-73 salt.

[0036] In any of the present methods, the dosage form administered can be an extended dosage form as described herein.

[0037] In another aspect, the present disclosure encompasses a pharmaceutical composition for treating a neurodegenerative disease, comprising an anti-neurodegenerative effective amount of A2-73. The therapeutically effective amount can range from about 0.5 mg to about 20 mg, from about 1 mg to about 60 mg, from about 30 mg to about 50 mg, or from about 3 mg to about 5 mg.

[0038] In another aspect, the disclosure encompasses a dosage form comprising an anti-neurodegeneratively effective amount of A2-73 effective for treating a neurodegenerative disease. The anti-neurodegeneratively effective amount of A2-73 can be from about 0.01 to about 10 mg / kg or from about 0.01 to about 10 mg / kg.

[0039] In one embodiment, the present disclosure encompasses a method for treating a neurodegenerative disease in a subject in need thereof. The method comprises administering an anti-neurodegenerative effective amount of A2-73 to the subject. The degenerative disease may be Alzheimer's disease, Parkinson's disease, prion disease, Huntington's disease, motor neuron disease (MND) such as amyotrophic lateral sclerosis, spinocerebellar ataxia (SCA), or spinal muscular atrophy (SMA).

[0040] An anti-neurodegenerative effective amount of A2-73 can be about 0.5 mg / day to about 100 mg / day, about 1 to about 60 mg / day, about 20 to about 50 mg / day, about 20 to about 30 mg / day, or about 15 to about 25 mg / day. Additionally, administering an anti-neurodegenerative effective amount of A2-73 to a subject can provide a blood level of A2-73 of about 10 ng / ml or about 12 ng / ml. [Brief explanation of the drawings]

[0041] [Figure 1] 1 is an overlay of XRPD patterns for Anavex 2-73 (hydrochloride salt) solid form. [Figure 2] 1 shows a micrograph of Form I crystals in polarized light. [Figure 3] 1 is a micrograph of Form I crystals obtained by sublimation. [Figure 4] 1 shows the XRPD pattern of Anavex 2-73 Form I derived from single crystal results for copper Kα radiation versus the experimentally measured XRPD pattern obtained for an isolated bulk sample of Form I. [Figure 5] Polarized light microscopy (PLM) of Form II. [Figure 6]1 shows the XRPD pattern of Form II obtained using copper Kα radiation versus the XRPD pattern from a single crystal. [Figure 7] Polarized light microscopy (PLM) of Form III. [Figure 8] 1 shows the XRPD pattern of Form III obtained using copper Kα radiation versus the XRPD pattern from a single crystal. [Figure 9] 1 shows the XRPD pattern of Form IV obtained using copper Kα radiation. [Figure 10] 1 shows the XRPD pattern of Form V obtained using copper Kα radiation. [Figure 11] 1 shows the XRPD pattern of Form VI obtained using copper Kα radiation. [Figure 12] 1 shows the XRPD pattern of Form VII obtained using copper Kα radiation. [Figure 13] Polarized light microscopy (PLM) of Form VIII. [Figure 14] 1 shows the XRPD pattern of Form VIII obtained using copper Kα radiation versus the XRPD pattern from a single crystal. [Figure 15] Non-polarized light microscopy (PLM) of free base form I. [Figure 16] XRPD pattern of Anavex2-73 free base Form I obtained using copper Kα radiation. [Figure 17] Polarized light microscopy (PLM) of Anavex2-73 sulfate form I. [Figure 18] XRPD pattern of Anavex2-73 sulfate Form I obtained using copper Kα radiation. [Figure 19] XRPD pattern of Anavex2-73 sulfate form II. [Figure 20] XRPD pattern of Anavex2-73 mesylate form I. [Figure 21] XRPD pattern of Anavex2-73 oxalate form I. [Figure 22] XRPD pattern of Anavex2-73 oxalate form II. [Figure 23]XRPD pattern of Anavex2-73 oxalate form III. [Figure 24] Polarized light microscopy (PLM) of Anavex2-73 dihydrogen phosphate form I. [Figure 25] XRPD pattern of Anavex2-73 dihydrogen phosphate form I obtained using copper Kα radiation. [Figure 26] XRPD pattern of A2-73 edisylic acid form I obtained using copper Kα radiation. [Figure 27] XRPD pattern of A2-73 benzoate salt Form I obtained using copper Kα radiation. [Figure 28] Polarized light microscopy (PLM) of Anavex2-73 hydrogen fumarate form I. [Figure 29] XRPD pattern of A2-73 hydrogen fumarate form I obtained using copper Kα radiation. [Figure 30] XRPD pattern of A2-73 hydrogen fumarate form II obtained using copper Kα radiation. [Figure 31] Polarized light microscopy (PLM) of Anavex2-73 hydrogen fumarate form III. [Figure 32] XRPD pattern of Anavex 2-73 hydrogen fumarate form III obtained using copper Kα radiation. [Figure 33] XRPD pattern of Anavex 2-73 hydrogen fumarate form IV obtained using copper Kα radiation. [Figure 34] XRPD pattern of Anavex 2-73 hydrogen fumarate form V obtained using copper Kα radiation. [Figure 35A] Sig-1R activation enhances autophagic activity. Figure 35A shows a Western blot and plots quantifying the results of a Western blot of autophagic flux upon addition of ANAVEX2-73. [Figure 35B] Figure 35B is a Western blot and plot quantifying the results of the Western blot of autophagic flux upon addition of PRE-084. Statistics are shown as mean + / - SD. ***p<0.001, **p<0.01, t-test, n=4. [Figure 35C] Figure 35C illustrates representative confocal fluorescence microscopy images and plots quantifying puncta in HEK293 cells stably transfected with a GFP-LC3B reporter construct (scale bar = 20 μm or 10 μm, respectively. 30 cells per treatment in three independent experiments. ***p<0.001, t-test). [Figure 36A] Sig-1R activation stimulates ULK1 activation and affects the expression levels of different autophagy network factors. Figure 36A shows a Western blot of ULK1 phosphorylation at serine 555 (pS555) upon treatment of HeLa cells with ANAVEX2-73, and a plot quantifying the Western blot results. Statistics are shown as mean ± SD. **p<0.01, t-test, n=4. [Figure 36B] Figure 36B shows a Western blot of ULK1 phosphorylation at serine 555 (pS555) upon treatment of HeLa cells with PRE-084. Statistics are shown as mean ± SD. *p<0.05, t-test, n=4. [Figure 36C] Figure 36C is a plot illustrating the relative expression levels of autophagy network factors analyzed using the autophagy qPCR array. The expression of each gene is plotted relative to control cells (set at 1), and the thresholds for up-regulation or down-regulation are defined as 1.5 and 0.67, respectively. [Figure 37A] Sig-1R activation by ANAVEX2-73 enhances autophagy in C. elegans. Figure 37A shows a Western blot and plot quantifying the results of a Western blot of GFP-LGG1 after treating worms with ANAVEX2-73. Statistics are shown as mean ± SD. *p<0.05, t-test, n=3. [Figure 37B]Figure 37B shows representative confocal fluorescence microscopy images of C. elegans treated with ANAVEX2-73 and BafiA1 or DMSO, and a plot quantifying the number of puncta in the microscopy images. Scale bar = 50 and 25 gm. Arrows indicate autophagosome structures. GFP-positive puncta + BafiA1 were compared to puncta in the control. ***p<0.001, t-test, and autophagic flux was calculated as indicated. [Figure 38A] Sig-1R activation by ANAVEX2-73 increases proteostatic capacity and ameliorates Aβ42-induced paralysis in C. elegans. Figure 38A shows representative confocal fluorescence microscopy images of Thioflavin S-positive Aβ42 aggregates in the head region of nematodes. Scale bar = 50 μm. [Figure 38B] Figure 38B is a plot of the analysis of Aβ42-induced paralysis. Statistics were performed using the log-rank test. Three independent experiments with a total of approximately 70 worms per treatment. Black = control, light gray = 50 μM ANAVEX2-73, dark gray = 100 μM ANAVEX2-73. DETAILED DESCRIPTION OF THE INVENTION

[0042] The present disclosure is based, in part, on the surprising discovery that crystalline polymorphs of tetrahydro-N,N-dimethyl-2,2-diphenyl-3-furanmethanamine ("A2-73" or "Anavex 2-73"), in free base or pharmaceutically acceptable salt form, are suitable for oral, transdermal, subcutaneous, or other administration and can be formulated to provide immediate or extended release of A2-73 upon administration. Crystalline polymorphs of A2-73, dosage forms, and formulations containing crystalline polymorphs of A2-73 are described below. Methods of using crystalline polymorphs of A2-73 for therapy are also disclosed, including the use of A2-73 for neuroprotection, including the treatment of neurodegenerative diseases.

[0043] I. Polymorphism In one aspect, the present disclosure provides crystalline polymorphs of A2-73. Each crystalline polymorph can be in free base form or salt form. The crystalline polymorphs are characterized by XRPD and other data provided herein. The properties of each crystalline polymorph are described below.

[0044] Form I, hydrochloride Form I is anhydrous, crystalline (birefringent plates and plate fragments) and has been shown via single crystal X-ray analysis to form racemic crystals, meaning that each individual crystal contains both enantiomers. Form I is the thermodynamically favored racemic crystalline form of the hydrochloride salt, the current Anavex 2-73 active pharmaceutical ingredient (API). Figure 2 shows polarized light microscopy (PLM) of Form I. Figure 3 shows crystals of Form I obtained by sublimation.

[0045] Single crystal X-ray analysis overview: Crystal type: Racemic Space group: Monoclinic P21 / c Unit cell parameters: a=14.1623(4)Å α=90.00° b=9.0974(3)Å β=102.103(3)° c=13.4052(4)Å γ=90.00° Volume = 1688.73(9) Å3 Z=4,Z'=1 Density (ρ calculated value) = 1.250g / cm3

[0046] The XRPD pattern of Anavex 2-73 Form I derived from single crystal results for copper Kα radiation versus the experimentally measured XRPD pattern obtained for an isolated bulk sample of Form I is shown in FIG.

[0047] Twenty best results for Anavex 2-73 Form I measured using copper Kα radiation The most intense XRPD peaks are shown in Table 1. [Table 1]

[0048] b. Form II, hydrochloride Form II is a crystalline hydrate (approximately monohydrate) consisting of columnar (rod-like) crystals. Single-crystal X-ray analysis indicates that Form II is a conglomerate consisting of homochiral crystals (meaning a physical mixture of optically pure crystals of the individual enantiomers). Form II is slightly hygroscopic; although the single-crystal results are modeled based on 0.8 moles of water per mole of Anavex 2-73, the crystal lattice can potentially hold up to 1.75 moles of water per mole of Anavex 2-73, making Form II likely a variable hydrate and undesirable from a pharmaceutical development perspective. Figure 5 shows polarized light microscopy (PLM) of Form II.

[0049] Single crystal X-ray analysis overview: Crystal type: homochiral Space group:Orthorhombic P212121 Unit cell parameters: a=7.10738(5)Å α=90.00° b=14.22620(10)Å β=90.00° c=17.18510(10)Å γ=90.00°volume=1737.608(16)Å 3 Z=4,Z'=1 Density (ρ 計算値 )=1.173g / cm 3

[0050] The XRPD pattern of Anavex 2-73 Form II derived from single crystal results for copper Kα radiation versus the experimentally measured XRPD pattern obtained for an isolated bulk sample of Form II is shown in FIG.

[0051] The 20 most intense XRPD peaks for Anavex 2-73 Form II measured using copper Kα radiation are shown in Table 2. [Table 2]

[0052] c. Form III, hydrochloride Form III is a slightly hygroscopic, anhydrous crystalline material that exhibits a columnar (rod-like) morphology. In some embodiments, single crystal X-ray analysis indicates that Form III is an optically pure form. In other embodiments, single crystal X-ray analysis indicates that Form III is a homochiral crystal (individual Form III appears to be a thermodynamically favored, optically pure form of Anavex 2-73. Figure 7 shows polarized light microscopy (PLM) of Form III.

[0053] Single crystal X-ray analysis overview: Crystal type: homochiral ·Space group:Orthorhombic P212121 Unit cell parameters: a=7.10738(5)Å α=90.00° b=14.22620(10)Å β=90.00° c=17.18510(10)Å γ=90.00° Volume = 1737.608(16) Å3 Z=4,Z'=1 Density (ρ calculated value) = 1.215g / cm3

[0054] XRPD pattern of Anavex2-73 Form III derived from single crystal results of copper Kα radiation versus experimentally measured XRPD pattern obtained for an isolated bulk sample of Form III (FIG. 8).

[0055] The 20 most intense XRPD peaks for Anavex 2-73 Form III measured using copper Kα radiation are shown in Table 3. [Table 3]

[0056] d. Form IV, hydrochloride Form IV is crystalline and can be isolated as a physical mixture with amorphous material during lyophilization of Anavex 2-73 from water. The XRPD pattern of Anavex 2-73 Form IV obtained using copper Kα radiation is shown in Figure 9.

[0057] The 11 most intense XRPD peaks for Anavex 2-73 Form IV measured using copper Kα radiation are shown in Table 4. [Table 4]

[0058] e. Form V, hydrochloride Form V is crystalline and can be isolated upon rotary evaporation of Anavex 2-73 from dichloromethane. The XRPD pattern of Anavex 2-73 Form V obtained using copper Kα radiation can be seen in Figure 10. The 20 most intense XRPD peaks of Anavex 2-73 Form V measured using copper Kα radiation are shown in Table 5. [Table 5]

[0059] f. Form VI, Hydrochloride Form VI is crystalline and was isolated by rapidly cooling an aqueous solution of Anavex 2-73 to 5° C. The XRPD pattern of Anavex 2-73 Form VI obtained using copper Kα radiation can be seen in FIG.

[0060] The 20 most intense XRPD peaks of Anavex 2-73 Form VI measured using copper Kα radiation are shown in Table 6. [Table 6]

[0061] g. Form VII, Hydrochloride Form VII is both crystalline and anhydrous and was isolated via air evaporation of Anavex 2-73 from methanol. The XRPD pattern of Anavex 2-73 Form VII obtained using copper Kα radiation is shown in Figure 12. The 20 most intense XRPD peaks for Anavex 2-73 Form VII measured using copper Kα radiation are shown in Table 7. [Table 7]

[0062] h. Form VIII, Hydrochloride Form VIII is a trihydrate crystalline form of Anavex 2-73. Single crystal X-ray analysis indicates that Form VIII consists of racemic crystals, meaning that each individual crystal contains both enantiomers. Without being bound by any particular theory, Form VIII is believed to be a layer or channel hydrate, in which the water of hydration is weakly associated and easily removed by grinding, drying, etc. The resulting material, a dehydrated lattice, is unstable and rapidly collapses to Form I. Figure 13 shows polarized light microscopy (PLM) of Form VIII.

[0063] Single crystal X-ray analysis overview: Crystal type: Racemic Space group: Monoclinic P21 / c Unit cell parameters: a=17.7753(11)Å α=90.00° b=9.0306(4)Å β=101.535(5)° c=13.2638(5)Å γ=90.00° Volume = 2086.12(12) Å 3 Z=4,Z'=1 density(ρ 計算値 )==1.184g / cm 3

[0064] The XRPD pattern of Anavex 2-73 Form VIII derived from a single crystal result with copper Kα radiation versus the experimentally measured XRPD pattern obtained for an isolated bulk sample of Form VIII is shown in Figure 14. The 20 most intense XRPD peaks for Anavex 2-73 Form VIII measured using copper Kα radiation are shown in Table 8. [Table 8]

[0065] i. Form I, free base A2-73 free base Form I is crystalline by XRPD and PLM, exhibiting highly birefringent aggregates of columnar (rod-like) crystals. Thermogravimetric analysis (TGA) showed no significant weight loss until after melting, indicating that Form I is anhydrous. This was confirmed by gravimetric vapor sorption (GVS) analysis, which showed minimal water renewal (0.3 wt.%) up to 90% RH. XRPD analysis after GVS showed no change in morphology. Differential scanning calorimetry (DSC) shows a sharp melting endotherm with an onset of approximately 89°C (peak temperature at approximately 91°C). Upon further heating, Form I appears to begin sublimating above approximately 120°C, with approximately 99% weight loss observed at 212.6°C. 1 The H NMR spectrum and HPLC-MS results were consistent with the structure of A2-73 free base. A2-73 free base was 99.9% and CAD analysis confirmed the absence of chloride in the sample. Figure 15 is a non-polarized light microscopy (PLM) of free base Form I. The XRPD pattern of Anavex2-73 free base Form I obtained using copper Kα radiation is shown in Figure 16.

[0066] The 20 most intense XRPD peaks for Anavex 2-73 free base Form I measured using copper Kα radiation are shown in Table 9. [Table 9]

[0067] Solubility: A2-73 free base surprisingly exhibited high solubility in all solvents tested except water, as shown in Table 10 below. [Table 10]

[0068] j. Sulfate Form I A2-73 sulfate Form I is crystalline and melts with a DSC onset temperature of approximately 184° C. Form I consisted of highly birefringent crystals. Its PLM and XRPD patterns are provided below.

[0069] Figure 17 is a polarized light microscopy (PLM) of Anavex 2-73 sulfate Form I. The XRPD pattern of Anavex 2-73 sulfate Form I obtained using copper Kα radiation is shown in Figure 18. The 20 most intense XRPD peaks for Anavex 2-73 sulfate Form I measured using copper Kα radiation are shown in Table 11. [Table 11]

[0070] k. Sulfate Form II A2-73 sulfate Form II is crystalline and melts with a DSC onset temperature of approximately 190° C. Form II appears to be metastable, converting to A2-73 sulfate Form I after storage at 40° C. / 75% RH. Its XRPD pattern, obtained using copper Kα radiation, is shown in Figure 19. The 20 most intense XRPD peaks for Anavex 2-73 sulfate Form II measured using copper Kα radiation are shown in Table 12. [Table 12]

[0071] l. Mesylate Form I A2-73 mesylate Form I is crystalline and melts with a DSC onset temperature of about 159° C. The XRPD pattern obtained using copper Kα radiation is shown in FIG.

[0072] The 20 most intense XRPD peaks for Anavex 2-73 mesylate Form I measured using copper Kα radiation are shown in Table 13. [Table 13]

[0073] m. Oxalate Form I A2-73 oxalate Form I is crystalline. Its XRPD pattern, obtained using copper Kα radiation, is provided in Figure 21. The 20 most intense XRPD peaks for Anavex 2-73 oxalate Form I measured using copper Kα radiation are listed in Table 14. [Table 14]

[0074] n.Oxalate Form II A2-73 oxalate Form II is crystalline. Its XRPD pattern, obtained using copper Kα radiation, is provided in Figure 22. The 20 most intense XRPD peaks for Anavex 2-73 oxalate Form II measured using copper Kα radiation in Table 15. [Table 15]

[0075] o.Oxalate Form III A2-73 oxalate Form III is anhydrous, crystalline, and melts with a DSC onset temperature of about 154° C. Its XRPD pattern, obtained using copper Kα radiation, is provided in FIG.

[0076] The 20 most intense XRPD peaks for Anavex 2-73 oxalate Form III measured using copper Kα radiation. [Table 16]

[0077] p. Dihydrogen phosphate form I A2-73 dihydrogen phosphate (mono-A2-73 phosphate) Form I is crystalline, hygroscopic, and melts with a DSC onset temperature of approximately 187-193°C. Isolated samples of Form I consist of small agglomerates of highly birefringent crystals and exhibit solubilities of approximately 47.2 and 33.1 mg / mL at pH 1.2 and 4.5, respectively. The pH of a saturated aqueous solution of Form I is 2.66. Its PLM and XRPD patterns are provided in Figures 24 and 25, respectively.

[0078] FIG. 24 is a polarized light microscopy (PLM) of Anavex 2-73 dihydrogen phosphate Form I.

[0079] The XRPD pattern of Anavex 2-73 dihydrogen phosphate Form I obtained using copper Kα radiation is shown in FIG.

[0080] The 20 most intense XRPD peaks of Anavex2-73 dihydrogen phosphate form I measured using copper Kα radiation. [Table 17]

[0081] q. Edisylate Form I A2-73 edisylic acid Form I is crystalline. Its XRPD pattern, obtained using copper Kα radiation, is provided in Figure 26.

[0082] The 20 most intense XRPD peaks of Anavex2-73 edisylate form I measured using copper Kα radiation. [Table 18]

[0083] r. Benzoate Form I A2-73 benzoate Form I is crystalline and melts with a DSC onset temperature of about 116° C. Its XRPD pattern, obtained using copper Kα radiation, is provided in FIG.

[0084] The 20 most intense XRPD peaks of Anavex 2-73 benzoate Form I measured using copper Kα radiation. [Table 19]

[0085] s. Fumaric acid hydrogen form I A2-73 hydrogen fumarate (mono-A2-73 fumarate) Form I is anhydrous, crystalline, and melts with a DSC onset temperature of approximately 193° C. Form I consists of small agglomerates of highly birefringent crystals. Its PLM and XRPD patterns are provided in Figures 28 and 29, respectively.

[0086] FIG. 28 shows polarized light microscopy (PLM) of Anavex 2-73 hydrogen fumarate Form I.

[0087] The XRPD pattern of Anavex 2-73 hydrogen fumarate Form I obtained using copper Kα radiation I is shown in FIG.

[0088] The 20 most intense XRPD peaks for Anavex 2-73 hydrogen fumarate form I measured using copper Kα radiation. [Table 20]

[0089] t. Fumaric acid hydrogen form II A2-73 fumarate salt Form II is crystalline and melts with a DSC onset temperature of about 196° C. Its XRPD pattern, obtained using copper Kα radiation, is provided in FIG.

[0090] The 17 most intense XRPD peaks for Anavex 2-73 fumarate Form II measured using copper Kα radiation. [Table 21]

[0091] u. Fumaric acid hydrogen form III A2-73 hydrogen fumarate (mono-A2-73 fumarate) Form III is anhydrous, crystalline, melts at a DSC onset temperature of approximately 197°C, is slightly hygroscopic, and picks up approximately 0.9 w / w% at 90% RH. Form III consists of highly birefringent prismatic (lathe-shaped) crystals. Form III exhibits moderate solubility at gastric pH (generally, approximately 1.5-3.5) and lower solubility at GI pH. GI pH refers to approximately pH 6 in the duodenum, gradually increasing to approximately pH 7.4 in the terminal ileum, then decreasing to approximately pH 5.7 in the cecum and gradually increasing to approximately pH 6.7 in the rectum. Measured solubilities of approximately 43.0, 4.1, and 12.6 mg / mL were observed for Form III at pH 1.2, 4.5, and 6.8, respectively. The pH of a saturated solution of Form III is 3.61. The PLM and XRPD patterns are provided in Figures 31 and 32.

[0092] FIG. 31 shows polarized light microscopy (PLM) of Anavex 2-73 hydrogen fumarate Form III.

[0093] The XRPD pattern of Anavex 2-73 hydrogen fumarate Form III obtained using copper Kα radiation is shown in FIG.

[0094] The 20 most intense XRPD peaks for Anavex 2-73 fumarate Form III measured using copper Kα radiation. [Table 22]

[0095] v. Fumarate Form IV A2-73 fumarate salt Form IV is anhydrous, crystalline, and melts with a DSC onset temperature of approximately 170° C., followed by sublimation. Its XRPD pattern, obtained using copper Kα radiation, is provided in FIG.

[0096] The 20 most intense XRPD peaks for Anavex 2-73 form fumarate IV measured using copper Kα radiation. [Table 23]

[0097] w. Fumarate Form V A2-73 fumarate salt (di-A2-73 fumarate salt) Form V is crystalline. Its XRPD pattern, obtained using copper Kα radiation, is provided in Figure 34.

[0098] The 20 most intense XRPD peaks for Anavex2-73 form V measured using copper Kα radiation. [Table 24]

[0099] II. Pharmaceutical Preparations One aspect of the present disclosure encompasses a pharmaceutical formulation for the delivery of A2-73, comprising a therapeutically effective amount of a crystalline form of A2-73 selected from A2-73 free base and any pharmaceutically acceptable salt of A2-73, as disclosed herein.

[0100] Pharmaceutical formulations can be prepared for extended or sustained release, or substantially immediate release, as known in the art, and can contain from about 1 mg to about 50 g of crystalline A2-73. For example, formulations for immediate delivery can contain a crystalline A2-73 salt, such as the hydrochloride salt. In other embodiments, formulations can be prepared for extended release of crystalline A2-73. In a non-limiting example, a formulation for extended release of crystalline A2-73 can contain crystalline A2-73 free base.

[0101] The pharmaceutical formulation further comprises one or more pharmaceutically acceptable non-medicinal ingredients. Non-limiting examples of non-medicinal ingredients include chemical enhancers, humectants, pressure-sensitive adhesives, antioxidants, solubilizers, thickeners, plasticizers, adjuvants, carriers, excipients, vehicles, coatings, and any combination thereof. The one or more non-medicinal ingredients may be present in a pharmaceutical composition for oral administration. The formulation may be selected for transdermal, parenteral, intraperitoneal, intravascular, subcutaneous, by inhalation spray, rectal, or pulmonary administration.

[0102] Crystalline A2-73 can generally be formulated to improve patient compliance and prevent the subject from removing the drug delivery device. For example, the formulation can be formulated to improve patient compliance and prevent the subject from removing the drug delivery device by providing a formulation for extended delivery. Extended delivery can range from more than one day to several months. This can be particularly relevant for patients with impaired cognitive and / or motor control abilities. Extended delivery can range from about one day to about one year, about one day to about one week, about three days to about one month, about two weeks to about six months, or about two months to about four months.

[0103] Extended-release formulations can be used for substantially continuous delivery of a drug at a preselected rate. For example, for crystalline A2-73, the drug can be delivered at a rate of about 1 mg to about 100 mg / day, about 40 to about 60 gm / day, or about 10 to about 30 gm / day. The appropriate amount of crystalline A2-73 can be readily determined by one of skill in the art based on, for example, the intended duration of administration of the drug via the extended-release formulation, the delivery mechanism, the particular formulation, and the relative potency of the drug, among other factors.

[0104] i. Binder Non-limiting examples of binders suitable for the various embodiments of the formulation include starch, pregelatinized starch, gelatin, polyvinylpyrrolidone, cellulose, methylcellulose, sodium carboxymethylcellulose, ethylcellulose, polyacrylamide, polyvinyloxoazolidone, polyvinyl alcohol, C 12 -C 18 These include fatty acid alcohols, polyethylene glycols, polyols, sugars, oligosaccharides, polypeptides, oligopeptides, and combinations thereof. Polypeptides can be any sequence of amino acids ranging from about 100 to about 300,000 daltons.

[0105] The binder may be introduced into the mixture to be granulated in a solid form, including but not limited to, crystals, particles, powder, or any other finely divided solid form known in the art. Alternatively, the binder may be dissolved or suspended in a solvent and sprayed onto the mixture in the granulation device as a binder fluid during granulation.

[0106] ii. Diluent Non-limiting examples of diluents (also referred to as "fillers" or "thinners") include carbohydrates, inorganic compounds, and biocompatible polymers such as polyvinylpyrrolidone (PVP). Other non-limiting examples of diluents include dibasic calcium sulfate, tribasic calcium sulfate, starch, calcium carbonate, magnesium carbonate, microcrystalline cellulose, dibasic calcium phosphate, tribasic calcium phosphate, magnesium carbonate, magnesium oxide, calcium silicate, talc, modified starches, sugars such as sucrose, dextrose, lactose, microcrystalline cellulose, fructose, xylitol, and sorbitol, polyhydric alcohols, starches, pre-fabricated direct compression diluents, and mixtures of any of the foregoing.

[0107] iii. Disintegrants Disintegrants can be effervescent or non-effervescent.Non-limiting examples of non-effervescent disintegrants include starches such as corn starch, potato starch, pregelatinized and modified starches thereof, sweeteners, clays such as bentonite, microcrystalline cellulose, alginates, sodium starch glycolate, gums such as agar, guar, carob, karaya, pesitin, and tragacanth.Suitable effervescent disintegrants include, but are not limited to, sodium bicarbonate in combination with citric acid, and sodium bicarbonate in combination with tartaric acid. Examples include sodium bicarbonate.

[0108] iv. Preservatives Non-limiting examples of preservatives include, but are not limited to, ascorbic acid and its salts, ascorbyl palmitate, ascorbyl stearate, anoxomer, N-acetylcysteine, benzyl isothiocyanate, m-aminobenzoic acid, o-aminobenzoic acid, p-aminobenzoic acid (PABA), butylhydroxyanisole (BHA), butylhydroxytoluene (BHT), caffeic acid, canthaxanthin, alpha-carotene, beta-carotene, beta-carotene, beta-apocarotenoic acid, carnosol, Carvacrol, catechin, cetyl gallate, chlorogenic acid, citric acid and its salts, clove extract, coffee bean extract, p-coumaric acid, 3,4-dihydroxybenzoic acid, N,N'-diphenyl-p-phenylenediamine (DPPD), dilauryl thiodipropionate, distearyl thiodipropionate, 2,6-di-tert-butylphenol, dodecyl gallate, edetic acid, ellagic acid, erythorbic acid, sodium erythorbate, esculetin, esculin, 6-ethoxy-1,2-dihydro-2,2,4-Trimethylquinoline, ethyl gallate, ethyl maltol, ethylenediaminetetraacetic acid (EDTA), eucalyptus extract, eugenol, ferulic acid, flavonoids (e.g., catechin, epicatechin, epicatechin gallate, epigallocatechin (EGC), epigallocatechin gallate (EGCG), polyphenol epigallocatechin-3-gallate), flavones (e.g., apigenin, chrysin, luteolin), flavonols (e.g., dacisin, myricetin, denferro), flavanones, fraxetin, fumaric acid, gallic acid, gentian extract, gluconic acid, glycine, gum yu bok choy (gum guaiacum), hesperetin, alpha-hydroxybenzylphosphinic acid, hydroxycinnamic acid, hydroxyglutaric acid, hydroquinone, N-hydroxysuccinic acid, hydroxytrilosol, hydroxyurea, rice bran extract, lactic acid and its salts, lecithin, lecithin citrate, R-alpha-lipoic acid, lutein, lycopene, malic acid, maltol, 5-methoxytryptamine, methyl gallate, monoglyceride citrate, monoisopropyl citrate, morin, beta-naphthoflavone, nordihydroguaretic acid (NDGA), octyl gallate, oxalic acid, palmityl citrate, phenothiazine, phosphatidylcholine, phosphoric acid, phosphate salts, phytic acid, phytyl bichromel, pimento extract, gallic acid Propyl, polyphosphate, quercetin, trans-resveratrol, rosemary extract, rosmarinic acid, sage extract, sesamol, silymarin, sinapic acid, succinic acid, stearyl citrate, syringic acid, tartaric acid, thymol, tocopherols (i.e., alpha-, beta-, gamma-, and delta-tocopherol), tocotrienols (i.e., alpha-, beta-, gamma-, and delta-tocotrienol), tyrosol, vanillic acid, 2,6-di-tert-butyl-4-hydroxymethylphenol (i.e., Ionox 100), 2,4-(tris-3',5'-bi-tert-butyl-4'-hydroxybenzyl)-mesitylene (i.e., Ionox 330), 2,4,5-trihydroxybutyrophenone, ubiquinone, tertiary butylhydroquinone (TBHQ), thiodipropionic acid, trihydroxybutyrophenone, tryptamine, tyramine, uric acid, vitamin K and derivatives, vitamin Q10, wheat germ oil, zeaxanthin, or combinations thereof.

[0109] v. Flavor modifiers Suitable flavor modifiers include flavors, flavorings, sweeteners, etc. Flavorings include, but are not limited to, synthetic flavor oils, flavoring aromatics, and / or natural oils, extracts from plants, leaves, flowers, fruits, and combinations thereof. Other non-limiting examples of flavors include cinnamon oil, oil of wintergreen, peppermint oil, clover oil, hay oil, anise oil, eucalyptus, vanilla, citrus oils such as lemon oil, orange oil, grape and grapefruit oil, fruit essential oils including apple, peach, pear, strawberry, raspberry, cherry, plum, pineapple, and apricot.

[0110] Flavoring agents include, but are not limited to, cellulose hydroxypropyl ethers (HPC), such as Klucel®, Nisswo HPC, and PrimaFlo HP22, low-substituted hydroxypropyl ethers (L-HPC), Seppifilm-LC, Pharmacoat®, Metolose SR, Opadry YS, PrimaFlo, MP3295A, Benecel MP824, and Benecel Cellulose hydroxypropyl methyl ether (HPMC) such as MP843, methylcellulose polymers such as Methocel® and Metolose®, ethylcellulose (EC) and mixtures thereof such as E461, Ethocel®, Aqualon®-EC, Surelease, polyvinyl alcohol (PVA) such as Opadry AMB, hydroxyethylcellulose such as Natrosol®, carboxymethylcellulose and salts of carboxymethylcellulose (CMC) such as Aualon®-CMC, Kollicoat Polyvinyl alcohol and polyethyl glycol copolymers such as IR®, monoglycerides (Myverol), triglycerides (KLX), polyethylene glycol, modified food starch, acrylic polymers and mixtures of acrylic polymers with cellulose ethers such as Eudragit® EPO, Eudragit® RD100, and Eudragit® E100, Sepifilm such as a mixture of cellulose acetate phthalate, HPMC and stearic acid, cyclodextrin, and mixtures of these materials. In other embodiments, additional flavor masking agents contemplated are those described in U.S. Patent Nos. 4,851,226, 5,075,114, and 5,876,759, each of which is incorporated herein by reference in its entirety.

[0111] Non-limiting examples of sweeteners include glucose (corn syrup), dextrose, invert sugar, fructose, and mixtures thereof (when not used as a carrier), saccharin and its various salts such as sodium ene, dipeptide sweeteners such as aspartame, dihydrochalcone compounds, glycyrrhizin, Stevia rebaudiana (stevioside), chloro derivatives of sucrose such as sucralose, sugar alcohols such as sorbitol, mannitol, and sciritol, hydrogenated starch hydrolysates, and the synthetic sweetener 3,6-dihydro-6-methyl-1,2,3-oxathiazin-4-one-2,2-dioxide, particularly the potassium salt (acesulfame-K), as well as the sodium and calcium salts thereof.

[0112] vi. Lubricants and glidants Lubricant compositions can be utilized to lubricate the components forming the pharmaceutical composition. As glidants, lubricants facilitate the removal of solid dosage forms during the manufacturing process. Non-limiting examples of lubricants and glidants include magnesium stearate, calcium stearate, zinc stearate, hydrogenated vegetable oil, Sterotex, polyoxyethylene monostearate, talc, polyethylene glycol, sodium benzoate, sodium lauryl sulfate, magnesium lauryl sulfate, and light mineral oil. Pharmaceutical compositions will generally contain from about 0.01% to about 10% by weight of a lubricant. In some embodiments, pharmaceutical compositions will contain from about 0.1% to about 5% by weight of a lubricant. In further embodiments, pharmaceutical compositions will contain from about 0.5% to about 2% by weight of a lubricant.

[0113] vii. Dispersants Dispersing agents may include, but are not limited to, starch, alginic acid, polyvinylpyrrolidone, guar gum, kaolin, bentonite, purified wood cellulose, sodium starch glycolate, isomorphous silicates, and microcrystalline cellulose as a high hydrophilic lipophilic balance (HLB) emulsifier surfactant.

[0114] viii. Coloring agents Depending on the embodiment of the present disclosure, it may be desirable to include a coloring agent. Suitable color additives include, but are not limited to, Food, Drug, and Cosmetic Colors (FD&C), Drug and Cosmetic Colors (D&C), or External Drug and Cosmetic Colors (Ext. D&C). These colors or dyes, along with their corresponding lakes and certain natural and derived colorants, may be suitable for use in various embodiments of the present disclosure.

[0115] ix. pH modifiers Non-limiting examples of pH modifiers include citric acid, acetic acid, tartaric acid, malic acid, fumaric acid, lactic acid, phosphoric acid, sorbic acid, benzoic acid, sodium carbonate, and sodium bicarbonate.

[0116] (x) chelating agent Chelating agents may be included as non-medicinal ingredients to immobilize oxidized groups, including, but not limited to, metal ions, to inhibit oxidative degradation of the morphinan by these oxidized groups. Non-limiting examples of chelating agents include lysine, methionine, glycine, gluconate, polysaccharides, glutamate, aspartate, and disodium ethylenediaminetetraacetic acid (NaEDTA).

[0117] (xi) antibacterial agents Antimicrobial agents may be included as non-medicinal ingredients to minimize degradation of compounds according to the present disclosure by microbial agents, including but not limited to bacteria and fungi. Non-limiting examples of antimicrobial agents include parabens, chlorobutanol, phenol, calcium propionate, sodium nitrate, sodium nitrite, Na2EDTA, and sulfites, including but not limited to sulfur dioxide, sodium bisulfite, and potassium bisulfite.

[0118] (xii) controlled release polymer The release-controlling polymer can be included in various embodiments of the solid dosage pharmaceutical composition incorporating the compound according to the present disclosure. In one embodiment, the release-controlling polymer can be used as a tablet coating. In other embodiments, including but not limited to, bilayer tablets, the release-controlling polymer can be mixed with granules and other non-medicinal ingredients before forming tablets by known processes, including but not limited to, compression in a tablet mold. Suitable release-controlling polymers include, but are not limited to, hydrophilic polymers and hydrophobic polymers.

[0119] Suitable hydrophilic release-controlling polymers include, but are not limited to, cellulose acetate, cellulose diacetate, cellulose triacetate, cellulose ethers, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, microcrystalline cellulose, nitrocellulose, cross-linked starch, agar, casein, chitin, collagen, gelatin, maltose, mannitol, maltodextrin, pectin, pullulan, sorbitol, xylitol, polysaccharides, ammonium alginate, sodium alginate, calcium alginate, potassium alginate, propylene glycol alginate, carmellose sodium alginate, carmellose calcium, caramelose cellulose, and cellulose acetate. Genan, fucoidan, furcellaran, gum arabic, carrageen gum, gum ghatti, guar gum, karaya gum, locust bean gum, okra gum, tragacanth gum, scleroglucan gum, xanthan gum, hypnea, laminaran, acrylic polymers, acrylate polymers, carboxyvinyl polymers, copolymers of maleic anhydride and styrene, copolymers of maleic anhydride and ethylene, copolymers of propylene maleic anhydride or copolymers of isobutylene maleic anhydride, cross-linked polyvinyl alcohol and poly N-vinyl-2-pyrrolidone, diesters of polyglucans, polyacrylamide, polyacrylic acid, polyamide, polyethylene glycol Examples of suitable hydrogels include chol, polyethylene oxide, poly(hydroxyalkyl methacrylate), polyvinyl acetate, polyvinyl alcohol, polyvinyl chloride, polystyrene, polyvinylpyrrolidone, anionic and cationic hydrogels, and combinations thereof.

[0120] (xiii) Coating A solid dosage containing a compound according to the present disclosure may include a coating, which may control the release of the compound, function as a moisture barrier or buffer, or modify the pH. As used herein, a "control-releasing coating" or "controlled-release coating" is defined to mean a functional coating that may include, for example, at least one pH-independent polymer, pH-dependent polymer (e.g., an enteric or reverse enteric polymer), soluble polymer, insoluble polymer, lipid, lipid material, or a combination thereof. When applied to a dosage form, the coating may be slow (e.g., when applied to a normal-release matrix dosage form), even slower (e.g., when applied to a controlled-release matrix dosage form), or, when applied to an uncoated dosage form, modify the release rate of a compound according to the present disclosure. For example, a controlled-release coating may be designed such that, when applied to a dosage form, the dosage form in combination with the controlled-release coating may exhibit a release of a compound according to the present disclosure such as a "modified release," "controlled release," "sustained release," "extended release," "delayed release," "extended release," or combinations thereof. A "controlled release coating" may optionally include additional materials that may alter the function of the controlled release coating.

[0121] As used herein, the term "moisture barrier" refers to a barrier that inhibits or delays the absorption of moisture. The compounds of the present disclosure may be hygroscopic and therefore susceptible to degradation over time under high humidity conditions. The proportions of the moisture barrier components and the amount of moisture barrier optionally applied on the controlled-release coating or on the core typically result in the moisture barrier not falling within the USP definition and requirements of an enteric coating. Preferably, the moisture barrier may comprise an enteric and / or acrylic polymer, preferably an acrylic polymer, optionally a plasticizer, and a permeation enhancer. The permeation enhancer is a hydrophilic substance that allows water to enter without physically disrupting the coating. The moisture barrier may additionally comprise other conventional inactive non-medicinal ingredients, which may improve the processing of the extended-release formulation.

[0122] Coating and matrix materials that can be used in accordance with the present invention are those known in the art for use in controlled-release formulations, such as synthetic polymers of the polyvinyl type, e.g., polyvinyl chloride, polyvinyl acetate and its copolymers, polyvinyl alcohol, and polyvinylpyrrolidone, synthetic polymers of the polyethylene type, e.g., polyethylene and polystyrene, acrylic acid polymers, biopolymers or modified biopolymers, e.g., cellulose polymers, shellac, and gelatin, fats, oils, higher fatty acids and higher alcohols (i.e., acids and alcohols containing alkyl chains of at least 10 carbon atoms), e.g., aluminum monostearate, cetyl alcohol, hydrogenated beef tallow, hydrogenated castor oil, 12-hydroxystearyl alcohol, glyceryl mono- or dipalmitate, glyceryl mono-, di-, or tristearate, myristyl alcohol, stearic acid, stearyl alcohol, and polyethylene glycol, waxes, sugars, and sugar alcohols.

[0123] The pH buffering properties of the coating are enhanced by the use of compounds from the group of compounds commonly used in antacid preparations, e.g., magnesium oxide, hydroxide, or carbonate, aluminum or calcium hydroxide, carbonate or silicate, complex aluminum / magnesium compounds, e.g., Al2O3·6MgO·CO2·12H2O, (Mg6Al2(OH) 16 CO3 4H2O) , MgO·Al2O3·2SiO2.nH2O, aluminum bicarbonate coprecipitate or similar compounds, or other pharmaceutically acceptable pH buffering compounds, such as sodium, potassium, calcium, magnesium, and aluminum salts of phosphorus, carbon, citric acid, or other suitable weak, inorganic, or organic acids, or suitable organic bases, including basic amino acids, and salts or combinations thereof, may be incorporated into the coating material.

[0124] The pH-dependent coating serves to release the drug in the desired region of the gastrointestinal (GI) tract, for example, the stomach or small intestine. If a pH-independent coating is desired, the coating is designed to achieve optimal release regardless of pH changes in the environmental fluid, for example, the GI tract. When the coating is formulated to release the compound according to the present disclosure in the intestine (particularly the upper small intestine), the coating is often referred to as an "enteric coating." pH-dependent coatings may include, but are not limited to, acrylic acid polymers and copolymers, for example, polymers formed from acrylic acid, methacrylic acid, methyl acrylate, ammoniomethyl acrylate, ethyl acrylate, methyl methacrylate, and / or ethyl methacrylate (e.g., Eudragit™); cellulose polymers such as hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methyl cellulose, methyl cellulose, ethyl cellulose, cellulose acetate, cellulose acetate phthalate (CAP), cellulose acetate trimellitate, hydroxypropyl methyl cellulose phthalate, hydroxypropyl methyl cellulose succinate, and sodium carboxymethyl cellulose; vinyl polymers and copolymers such as shellac (purified lac), polyvinylpyrrolidone, polyvinyl acetate, polyvinyl acetate phthalate (PVAP), vinyl acetate chlorine phosphate copolymer, and ethylene-vinyl acetate copolymer; zein; and salts and combinations thereof.

[0125] A. Transdermal Administration One aspect of the present disclosure encompasses formulations of A2-73 for transdermal administration. Non-limiting examples of transdermal formulations include, but are not limited to, those used in transdermal patches, such as gels, ointments, emulsions, microemulsions, aqueous gels, foams, sprays, lotions, or creams.

[0126] In one embodiment, the transdermal formulation of the crystalline form of A2-73 is a transdermal patch. The transdermal patch comprises a therapeutically effective amount of the crystalline form of A2-73. The crystalline form of A2-73 can be A2-73 free base or an A2-73 salt.

[0127] The crystalline form of A2-73 in the patch can be A2-73 free base. When the crystalline form of A2-73 is A2-73 free base, the patch can contain about 40 mg to about 60 mg, about 80 mg to about 120 mg, or about 180 mg to about 220 mg of A2-73 free base. Alternatively, the crystalline form of A2-73 in the patch can be A2-73 fumarate. When the crystalline form of A2-73 is A2-73 fumarate, the patch can contain about 1 mg to about 55 mg of A2-73 fumarate.

[0128] Transdermal patches include those formulated for extended or sustained release and those formulated for substantially immediate release. For example, extended-release transdermal patches may contain crystalline forms of A2-73, such as the free base or A2-73 fumarate salt, as disclosed herein. For example, immediate-release patch forms may contain A2-73 salts, such as the HCl salt.

[0129] The transdermal patch may provide extended release of A2-73 over a period ranging from about 1 day to about 7 days. Additionally, the transdermal patch may provide a dose of about 250-350 μg / cm 2 Maximum transdermal flow in the range of / h A2-73 may be present.

[0130] Transdermal patch can be matrix patch or reservoir patch.In one embodiment, patch is matrix patch.The transdermal patch that contains the compound amount to be delivered in matrix or reservoir for the extended delivery of compound is known in the art, and can be as described in, for example, U.S. Patent No. 9,656,441 and U.S. Patent Publication No. 2019 / 0099383, the disclosures of which are incorporated herein in their entirety.

[0131] The matrix patch may be covered by a peripheral pressure-sensitive adhesive that extends beyond the patch in all directions. The patch may further contain one or more other non-medicinal ingredients, such as those described in Section III herein, and may be selected from chemical enhancers, humectants, pressure-sensitive adhesives, antioxidants, solubilizers, thickeners, plasticizers, and any combination thereof.

[0132] The matrix layer of the transdermal patch can be formulated for extended release. For example, in addition to containing a therapeutically effective amount of active ingredient, the matrix formulation can further contain one or more pharmaceutically acceptable carriers or non-medicinal ingredients. Non-limiting examples of pharmaceutically acceptable carriers or non-medicinal ingredients include chemical penetration enhancers (CPEs), chemical enhancers, wetting agents, pressure-sensitive adhesives, antioxidants, solubilizers, thickeners, plasticizers, and any combination thereof.

[0133] In some embodiments, the matrix layer comprises one or more CPEs, non-limiting examples of which include anionic surfactants, cationic surfactants, zwitterionic surfactants, nonionic surfactants, fatty acids, fatty acid esters, azone and azone-like compounds, ethanol, glycerol monolaurate, DMF, polyethylene glycol monolaurate, DMSO, ethyl alcohol, oleic acid, oleyl alcohol, glycerol monooleate, levulinic acid, dipropylene glycol, diethylene glycol monoethyl ether, lauric acid lactate, and combinations thereof.

[0134] In some embodiments, the transdermal patch of the present disclosure includes a matrix layer having an upper side and a lower side, the matrix containing a therapeutically effective amount of a crystalline form of A2-73 selected from A2-73 free base and A2-73 salt. The patch also includes an adhesive layer having an upper side and a lower side, the lower side of the adhesive layer contacting the upper side of the matrix layer, the adhesive layer having a first portion covering the upper side of the matrix layer and a second portion extending to the side of the matrix layer. The bottom side of the matrix contacts the user's skin.

[0135] The transdermal patch may further include a protective layer covering the underside of the matrix and the underside of the second portion of the adhesive layer. Furthermore, the transdermal patch may include a cover layer on the upper side of the matrix. Preferably, the cover layer is at least partially bielastic. For example, the bielastic cover layer may include an acrylic copolymer with hydroxyl functionality. In some cases, the transdermal patch may include a separation layer located between the upper side of the matrix layer and the lower side of the adhesive layer.

[0136] The surface area of ​​the matrix in contact with the subject's skin is approximately 1 cm 2 ~about 20cm 2 , about 3cm 2 ~about 5cm 2 , or about 8 cm 2 ~about 10cm 2 The range may be:

[0137] In one embodiment, the patch comprises a matrix containing either A2-73 free base or A2-73 fumarate, or a reservoir containing A2-73 free base or A2-73 fumarate. Other non-medicinal ingredients / chemicals / reagents that may be included in the patch matrix or reservoir include ethyl oleate (EO), Tween 60, Tween 4, 0, Tween 80, triethanolamine and ethanol, propylene glycol (PG) and polyvinyl alcohol (PVA), polyethylene glycol 400 (PEG 400), and methanol, or any combination thereof. Patch components may include a backing membrane (3M-9720), a rate-controlling membrane (3M-CoTran 9728 (2 mil) and 9716 (4 mil)), and a release liner (SCOTCHPAK 9755), an acrylate adhesive Duro-Tak 387 / 2510.

[0138] B. Oral Formulations Some embodiments of the present disclosure include oral formulations for delivering crystalline forms of A2-73 free base and salts. Oral formulations are known in the art and include, but are not limited to, tablets, including suspension tablets, chewable tablets, effervescent tablets, or caplets; pills; powders, such as sterile packaged powders, dispensable powders, and effervescent powders; capsules, including both soft and hard gelatin capsules, such as HPMC capsules; lozenges; sachets; sprinkles; reconstitutable powders or shakes; troches; pellets; granules; liquids; suspensions; emulsions; or semisolids and gels. Alternatively, pharmaceutical compositions can be incorporated into food or powders for mixing with liquid, or can be orally administered after mixing with non-food liquids only.

[0139] Oral dosage forms include those formulated for extended or sustained release and those formulated for substantially immediate release. For example, extended release oral dosage forms may contain crystalline forms of A2-73 as free base or A2-73 fumarate as disclosed herein. For example, immediate release oral dosage forms may contain A2-73 salts, such as HCl salts. Release characteristics and release times may be measured according to methods known in the art.

[0140] In one embodiment, an oral dosage form comprising crystalline A2-73 can provide extended release of A2-73 over a period ranging from about 1 day to about 3 days, 4 to 24 hours, e.g., 6 to 24 hours, preferably 12 to 24 hours, and can provide delivery of about 15 to about 30 mg / day of A2-73 to a subject.

[0141] In another embodiment, an oral dosage form comprising crystalline A2-73 provides substantially immediate release of A2-73 as understood in the art, and may include, for example, an immediate release oral dosage form of A2-73 hydrochloride.

[0142] In one embodiment, the oral formulation is an enteric-coated oral dosage form comprising a core matrix ("core") containing a therapeutically effective amount of a crystalline form of A2-73. The crystalline A2-73 can be A2-73 free base or an A2-73 salt. The core is surrounded by a coating. Preferably, the coating is an enteric coating.

[0143] The solid core, e.g., capsule or tablet formulations of the present disclosure contain crystalline A2-73 along with non-medicinal ingredients, non-limiting examples of which may be as described in Section III above and include binders, diluents (fillers), disintegrants, effervescent disintegration agents, preservatives (antioxidants), flavor modifiers, lubricants and glidants, dispersants, colorants, pH modifiers, chelating agents, antimicrobial agents, release-controlling polymers, and combinations of any of these agents.

[0144] Non-limiting examples of binders suitable for oral formulations include starch, pregelatinized starch, gelatin, polyvinylpyrrolidone, cellulose, methylcellulose, sodium carboxymethylcellulose, ethylcellulose, polyacrylamide, polyvinyloxoazolidone, polyvinyl alcohol, C 12 -C 18 These include fatty acid alcohols, polyethylene glycols, polyols, sugars, oligosaccharides, polypeptides, oligopeptides, and combinations thereof. Polypeptides can be any sequence of amino acids ranging from about 100 to about 300,000 daltons.

[0145] Non-limiting examples of diluents (also referred to as "fillers" or "thinners") include carbohydrates, inorganic compounds, and biocompatible polymers such as polyvinylpyrrolidone (PVP). Other non-limiting examples of diluents include dibasic calcium sulfate, tribasic calcium sulfate, starch, calcium carbonate, magnesium carbonate, microcrystalline cellulose, dibasic calcium phosphate, tribasic calcium phosphate, magnesium carbonate, magnesium oxide, calcium silicate, talc, modified starches, sugars such as sucrose, dextrose, lactose, microcrystalline cellulose, fructose, xylitol, and sorbitol, polyhydric alcohols, starches, pre-fabricated direct compression diluents, and mixtures of any of the foregoing.

[0146] Disintegrants can be effervescent or non-effervescent.Non-effervescent disintegrants include, but are not limited to, starches such as corn starch, potato starch, pregelatinized and modified starches thereof, sweeteners, clays such as bentonite, microcrystalline cellulose, alginates, sodium starch glycolate, gums such as agar, guar, carob, karaya, pesitin, and tragacanth.Suitable effervescent disintegrants include, but are not limited to, sodium bicarbonate in combination with citric acid and sodium bicarbonate in combination with tartaric acid.

[0147] Dispersing agents may include, but are not limited to, starch, alginic acid, polyvinylpyrrolidone, guar gum, kaolin, bentonite, purified wood cellulose, sodium starch glycolate, isomorphous silicates, and microcrystalline cellulose as a high hydrophilic lipophilic balance (HLB) emulsifier surfactant.

[0148] Non-limiting examples of pH modifiers include citric acid, acetic acid, tartaric acid, malic acid, fumaric acid, lactic acid, phosphoric acid, sorbic acid, benzoic acid, sodium carbonate, and sodium bicarbonate.

[0149] The release-controlling polymer can be included in the oral formulation incorporating the compound according to the present disclosure.In one embodiment, the release-controlling polymer can be used as a tablet coating.In other embodiments, including but not limited to, bilayer tablets, the release-controlling polymer can be mixed with granules and other non-medicinal ingredients before forming tablets by known processes, including but not limited to, compression in a tablet mold.Suitable release-controlling polymers include but are not limited to hydrophilic polymers and hydrophobic polymers.

[0150] The coating may control the release of the compound, function as a moisture barrier or buffer, or modify the pH. As used herein, "control-releasing coating" or "controlled-release coating" is defined to mean a functional coating that may include, for example, at least one pH-independent polymer, pH-dependent polymer (e.g., enteric or reverse enteric polymer), soluble polymer, insoluble polymer, lipid, lipid material, or a combination thereof. When applied to a solid dosage form, the coating may be slow (e.g., when applied to a normal-release matrix dosage form), or even slower (e.g., when applied to a controlled-release matrix dosage form), or when applied to an uncoated dosage form, may modify the release rate of a compound according to the present disclosure. For example, a controlled-release coating may be designed such that, when applied to a dosage form, the dosage form in combination with the controlled-release coating may exhibit a release of a compound according to the present disclosure such as "immediate release," "modified release," "controlled release," "sustained release," "extended release," "delayed release," "extended release," or a combination thereof. The coating may optionally include additional materials that may modify the function of the controlled release coating.

[0151] The pH buffering properties of the coating are enhanced by the use of compounds from the group of compounds commonly used in antacid preparations, e.g., magnesium oxide, hydroxide, or carbonate, aluminum or calcium hydroxide, carbonate or silicate, complex aluminum / magnesium compounds, e.g., Al2O3·6MgO·CO2·12H2O, (Mg6Al2(OH) 16 CO3·4H2O), MgO·Al2O3·2SiO2·nH2O, aluminum bicarbonate coprecipitate or similar compounds, or other pharmaceutically acceptable pH buffering compounds, such as sodium, potassium, calcium, magnesium, and aluminum salts of phosphorus, carbon, citric acid, or other suitable weak, inorganic, or organic acids, or suitable organic bases, including basic amino acids, and salts or combinations thereof, may be incorporated into the coating material.

[0152] The pH-dependent coating serves to release the drug in the desired region of the gastrointestinal (GI) tract, for example, the stomach or small intestine. If a pH-independent coating is desired, the coating is designed to achieve optimal release regardless of pH changes in the environmental fluid, for example, the GI tract. When the coating is formulated to release the compound according to the present disclosure in the intestine (particularly the upper small intestine), the coating is often referred to as an "enteric coating." pH-dependent coatings may include, but are not limited to, acrylic acid polymers and copolymers, for example, polymers formed from acrylic acid, methacrylic acid, methyl acrylate, ammoniomethyl acrylate, ethyl acrylate, methyl methacrylate, and / or ethyl methacrylate (e.g., Eudragit™); cellulose polymers such as hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methyl cellulose, methyl cellulose, ethyl cellulose, cellulose acetate, cellulose acetate phthalate (CAP), cellulose acetate trimellitate, hydroxypropyl methyl cellulose phthalate, hydroxypropyl methyl cellulose succinate, and sodium carboxymethyl cellulose; vinyl polymers and copolymers such as shellac (purified lac), polyvinylpyrrolidone, polyvinyl acetate, polyvinyl acetate phthalate (PVAP), vinyl acetate chlorine phosphate copolymer, and ethylene-vinyl acetate copolymer; zein; and salts and combinations thereof.

[0153] Coating and core materials that may be used in accordance with the present invention are those known in the art for use in controlled-release formulations, such as synthetic polymers of the polyvinyl type, for example, polyvinyl chloride, polyvinyl acetate, and copolymers thereof, polyvinyl alcohol, and polyvinylpyrrolidone; synthetic polymers of the polyethylene type, for example, polyethylene and polystyrene; acrylic acid polymers; biopolymers or modified biopolymers, for example, cellulose polymers, shellac, and gelatin; fats, oils, higher fatty acids, and higher alcohols (i.e., acids and alcohols containing alkyl chains of at least 10 carbon atoms), for example, aluminum monostearate, cetyl alcohol, hydrogenated tallow, hydrogenated castor oil, 12-hydroxystearyl alcohol, glyceryl mono- or dipalmitate; glyceryl mono-, di-, or tristearate; myristyl alcohol, stearic acid, stearyl alcohol, and polyethylene glycol; waxes; sugars and sugar alcohols.

[0154] The crystalline A2-73 in the core can be crystalline A2-73 free base, and the core can contain about 1 g to about 50 g of crystalline A2-73. The core can contain about 1 mg to about 50 mg of A2-73 free base. The core can also contain about 1 g to about 50 g, about 10 mg to about 50 mg, about 20 mg to about 30 mg, or about 15 mg to about 25 mg of A2-73 fumarate. The core can contain about 35% to about 40% by weight of A2-73 free base or A2-73 fumarate. In one embodiment, the core contains about 35% to about 40% by weight of A2-73 free base or A2-73 fumarate, about 55% to about 70% by weight of hydroxypropylmethylcellulose. The composition contains cellulose acetate succinate, about 0.3% to about 0.9% by weight of magnesium stearate, and about 0.05% to about 0.5% by weight of colloidal silicon dioxide. The hydroxypropyl methylcellulose acetate succinate is soluble in aqueous solution having a pH of about 5.5 or higher, the second grade hydroxypropyl methylcellulose acetate succinate is soluble in aqueous solution having a pH of about 6.8 or higher, and combinations thereof.

[0155] C. Subcutaneous The formulation may be a subcutaneously injectable dosage formulation. In some embodiments, the formulation may be an extended-release subcutaneously injectable dosage formulation or a substantially immediate-delivery formulation. The extended-release subcutaneous dosage formulation may contain from about 0.1 to about 5 g of crystalline A2-73 to about 0.5 g to about 3 g of crystalline A2-73.

[0156] Injectable dosage formulations for extended release of drugs are known in the art and may include injectable formulations formulated for extended delivery of drugs, such as implantable drug delivery devices. As used herein, the term "drug delivery device" refers to any implantable device suitable for delivering a formulation according to the present disclosure. Non-limiting examples of devices include any implantable device having any mechanism of action, including diffusive, erodible, or convective systems, such as osmotic pumps, biodegradable implants, electrodiffusion systems, electroosmotic systems, vapor pressure pumps, electrolytic pumps, effervescent pumps, piezoelectric pumps, erosion-based systems, or electromechanical systems.

[0157] III. Dosage form One embodiment of the present disclosure encompasses a dosage form of A2-73. The dosage form contains a therapeutically effective amount of A2-73 in crystalline form. For example, the dosage form may contain a neuroprotective amount of crystalline A2-73. In some embodiments, the neuroprotective amount is an anti-neurodegenerative amount of A2-73 in a crystalline form as disclosed herein. The crystalline A2-73 may be A2-73 free base or a crystalline A2-73 salt. The dosage form may be formulated as described in Section II above.

[0158] Dosage forms can contain about 1 mg to about 50 g, about 1 mg to about 500 mg, or about 1 mg to about 100 mg of A2-73 free base or A2-73 salt.

[0159] Dosage forms can contain about 1 mg to about 500 mg, about 50 to about 400 mg, about 75 to about 150 mg, or about 150 to about 200 mg of A2-73 free base or A2-73 salt. For example, dosage forms can contain 1, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 140, 160, 180, 200, 220, 240, 260, 280, or 300 mg or more of A2-73 free base or A2-73 salt. In some embodiments, dosage forms can contain about 1 mg to about 500 mg, or about 1 mg to about 100 mg of A2-73 free base or A2-73 salt.

[0160] In some embodiments, the crystalline form of A2-73 in the dosage form is the free base. When A2-73 is the free base, the dosage form can contain from about 1 mg to about 500 mg, from about 40 mg to about 60 mg, from about 80 mg to about 120 mg, or from about 180 mg to about 220 mg of A2-73 free base.

[0161] In other embodiments, the crystalline form of A2-73 in the dosage form is a salt of A2-73. When A2-73 is a salt, the dosage form may contain from about 1 mg to about 500 mg, from about 1 mg to about 55 mg, from about 40 mg to about 60 mg, from about 80 mg to about 120 mg, or from about 180 mg to about 220 mg of the A2-73 salt.

[0162] Dosage forms include those formulated for extended or sustained release and those formulated for immediate release. For example, an immediate-release dosage form may contain a crystalline form of A2-73 as the free base or an A2-73 salt, as disclosed herein. For example, a fast-disintegrating oral dosage form may contain an A2-73 salt, such as, for example, an HCl salt. Alternatively, a dosage form may contain a crystalline form of A2-73 as the free base or an A2-73 salt formulated for inhaled drug delivery, either as a dry powder or an aerosol spray.

[0163] Dosage forms also include those formulated for topical administration. For example, the dosage form may be formulated as one or more of a gel, ointment, emulsion, microemulsion, solution, suspension, paste, gel, foam, spray, lotion, or cream. In one embodiment, the topical administration dosage form is a transdermal patch. The transdermal patch may be, for example, as described in Section III below. When the dosage form is formulated as a transdermal patch, the transdermal patch may contain about 40 mg to about 60 mg, about 80 mg to about 120 mg, or about 180 mg to about 220 mg of A2-73 free base in crystalline form.

[0164] Alternatively, dosage form can be formulated for oral administration.The dosage form formulated for oral administration can be the tablet, capsule and chewable capsule, powder, granule, tea, drops, or liquid medicine or syrup that can be swallowed, chewed, or dissolved in water or under the tongue.Preferably, dosage form is enteric coated oral preparation.Enteric coated oral preparation can be as described in the following section IV.

[0165] When the dosage form is an enteric coated oral formulation, the formulation may contain about 0.1 mg to about 60 mg of A2-73 free base, preferably about 1 mg to about 50 mg of A2-73 free base.

[0166] The enteric-coated oral formulation may also contain a crystalline form of an A2-73 salt. The A2-73 salt may be a fumarate, sulfate, mesylate, dihydrogen phosphate, edisylate, benzoate, hydrochloride, or oxalate salt. In one embodiment, the A2-73 salt is a fumarate salt. When the A2-73 salt is a fumarate salt, the enteric-coated oral formulation may contain about 0.1 to about 100 mg of A2-73 fumarate, preferably about 1 mg to about 55 mg of A2-73 fumarate.

[0167] Dosage forms also include those formulated for subcutaneous and / or intramuscular injection. For example, intramuscular dosage forms may contain the free base form of A2-73 dissolved in an oil matrix for intramuscular injection, or alternatively, prepared as a suspension of the free base for intramuscular injection. Dosage forms formulated for subcutaneous or intramuscular injection may contain the salt or free base form of A2-73 disclosed herein prepared as microspheres using methods known in the art. Alternatively, the free base or salt form of A2-73 may be coated with a thin layer coating, such as a zinc oxide coating, using, for example, atomic layer deposition (ALD) techniques, and used in formulations for subcutaneous or intramuscular injection. Alternatively, A2-73 free base may be dissolved in a biodegradable polymer matrix and then implanted subcutaneously (or used in a transdermal patch, as further described below).

[0168] IV.A2-73 Administration and Treatment Methods One aspect of the present disclosure includes a method of administering A2-73 to a subject, the method comprising administering A2-73 to the subject in a dosage form comprising a crystalline form of A2-73 selected from A2-73 free base and A2-73 salts.

[0169] The dosage form may be as described in Section III above. The dosage form may be formulated for immediate or extended release, and may be formulated for oral, transdermal, subcutaneous, or other modes of administration. The formulation may be as described in Section II above.

[0170] Extended-release dosage forms containing crystalline A2-73 can administer A2-73 for a period of about 2 weeks, 30 days, about 45 days, about 60 days, about 90 days, or about 120 to about 180 days.

[0171] In some embodiments, the method includes administering a topical dosage form. The topical dosage form can be a transdermal patch. The transdermal patch can be replaced daily, weekly, or more frequently. In some embodiments, the transdermal patch can maintain a blood A2-73 level in the subject's blood for a period of time ranging from about 5 ng / ml to about 15 ng / ml, and particularly about 10 ng / ml.

[0172] The crystalline forms of A2-73 can also be administered orally using a dosage form formulated for oral administration. Preferably, the dosage form is an enteric-coated oral formulation.

[0173] The enteric-coated oral dosage form can be administered every other day. The dosage form can deliver about 15 to about 30 mg of A2-73 per day. Furthermore, when formulated for extended release of A2-73, the oral dosage form can deliver A2-73 for a period that can range from about 1 day to about 7 days, about 48 hours, about 72 hours, or longer.

[0174] One aspect of the present disclosure includes a method of treating Alzheimer's disease in a subject in need thereof, comprising administering a therapeutically effective amount of a dosage form comprising a crystalline form of A2-73 selected from A2-73 free base and A2-73 salt.

[0175] One aspect of the present disclosure includes a method of treating progressive dementia in a subject in need thereof, comprising administering a therapeutically effective amount of a dosage form comprising a crystalline form of A2-73 selected from A2-73 free base and A2-73 salt.

[0176] V. Treatment of Neurodegenerative Diseases Sig-1R expression or activity is associated with neurodegeneration, and Sig-1R activation is associated with neuroprotection in different in vitro and in vivo models using different types of pharmacological Sig-1R activators with different pharmacological profiles. The inventors have surprisingly discovered that the mixed muscarinic receptor ligand A2-73 and Sig-1R agonists can be used to treat neurodegenerative diseases. As such, the crystalline form of A2-73, as well as any of the disclosed topical and oral dosage forms, can be administered to a subject in need thereof for neuroprotection, including the treatment of neurodegenerative diseases.

[0177] As such, one aspect of the present disclosure includes a pharmaceutical composition for treating a neurodegenerative disease. The composition comprises an anti-neurodegenerative effective amount of A2-73. A2-73 may be a crystalline polymorph of A2-73, and may be the free base or a salt. Preferably, A2-73 is the hydrochloride salt of A2-73.

[0178] The effective anti-neurodegenerative amount can range from about 0.5 mg to about 20 mg, from about 1 mg to about 60 mg, from about 30 mg to about 50 mg, or from about 3 mg to about 5 mg.

[0179] Another embodiment of the present disclosure includes a dosage form comprising an amount of A2-73 effective for treating a neurodegenerative disease. The amount of A2-73 in the dosage form can be from about 0.01 to about 10 mg / kg.

[0180] Another aspect of the present disclosure includes a method of treating a neurodegenerative disease in a subject in need thereof, comprising administering to the subject an anti-neurodegenerative effective amount of A2-73. The disease may be selected from Alzheimer's disease, Parkinson's disease, prion diseases, Huntington's disease, motor neuron diseases (MND) such as amyotrophic lateral sclerosis, spinocerebellar ataxia (SCA), and spinal muscular atrophy (SMA).

[0181] An anti-neurodegenerative effective amount of A2-73 can range from about 0.5 mg / day to about 100 mg / day, from about 1 to about 60 mg / day, from about 20 to about 50 mg / day, from about 20 to about 30 mg / day, or from about 15 to about 25 mg / day. Administering an anti-neurodegenerative effective amount of A2-73 can provide blood levels of about 10 ng / ml, about 12 ng / ml, or about A2-73.

[0182] definition Unless otherwise defined, all technical and scientific terms used herein have the meaning commonly understood by one of ordinary skill in the art to which this invention belongs. The following references provide those skilled in the art with general definitions of many of the terms used in this invention: Singleton et al., Dictionary of Microbiology and Molecular Biology (2nd ed.1994), The Cambridge Dictionary of Science and Technology (Walker ed., 1988), The Glossary of Genetics, 5th Ed., R. Rieger et al. (eds.), Springer Verlag (1991), and Hale & Marham, The Harper. Collins Dictionary of Biology (1991). As used herein, the following terms have the meanings ascribed to them unless otherwise specified.

[0183] When introducing elements of the disclosure or preferred embodiment(s) thereof, the articles "a," "an," "the," and "said" are intended to mean that there are one or more of the elements. The terms "comprising," "including," and "having" are intended to be inclusive and mean that there may be additional elements other than the listed elements.

[0184] Since various changes can be made in the cells and methods described above without departing from the scope of the present invention, it is intended that all matter contained in the above description and in the examples given below should be interpreted in an illustrative and not a limiting sense.

[0185] The term "comprising" means "including, but not necessarily limited to," and specifically indicates open-ended inclusion or membership in such listed combinations, groups, series, etc. As used herein, the terms "comprising" and "including" are inclusive and / or open-ended and do not exclude additional, unrecited elements or method steps. The term "consisting essentially of" is more restrictive than "comprising," but less restrictive than "consisting of." Specifically, the term "consisting essentially of" limits membership to specified materials or steps, and to materials or steps that do not materially affect the essential characteristics of the claimed invention.

[0186] As used herein, the term "subject" refers to a mammalian subject, including, but not limited to, humans, non-human primates, mice, rats, guinea pigs, and dogs.

[0187] As used herein, the terms "extended" or "sustained" release or delivery are used interchangeably and can be understood in contrast to immediate release compositions. In extended release formulations, the active ingredient is gradually and continuously released over time at a rate appropriate for the intended use of the dosage form. In particular, the term refers to the fact that the formulation does not release the entire dose of the active ingredient immediately after administration, and that the formulation is administered more frequently. A sustained or extended release dosage form, used synonymously with long-acting, sustained release, or modified release, is a dosage form that allows for a reduction in dosing frequency or a significant increase in patient compliance or therapeutic performance compared to that presented as an immediate release dosage form (e.g., as a solution or conventional solid dosage form of immediate drug release). [Example]

[0188] The publications discussed above are provided solely for their disclosure prior to the filing date of the present application. Nothing herein should be construed as an admission that the present invention is not entitled to antedate such disclosure by virtue of prior invention.

[0189] The following examples are included to demonstrate the present disclosure. It should be understood by those skilled in the art that the techniques disclosed in the following examples represent techniques discovered by the inventors to work well in the practice of the present disclosure. However, those skilled in the art should understand in light of the present disclosure that many changes can be made in the present disclosure without departing from the spirit and scope of the present disclosure and still obtain the same or similar results, and therefore all matters described should be interpreted as illustrative and not limiting.

[0190] Example 1. Preparation of Form I, Hydrochloride Salt Form I can be obtained by crystallization of Anavex2-73 from an anhydrous solvent, such as isopropyl alcohol (IPA). At 70°C, Form I can be obtained, for example, from IPA containing up to at least 2.5% volume / volume water. Form I can also be obtained by sublimation. Some examples of Form I preparation are as follows:

[0191] (i) Example 1 Approximately 100 mg of Anavex 2-73 was weighed into a sample vial, to which 0.5 mL or 1 mL of 2-ethoxyethanol, 1-propanol, acetone, acetonitrile, dichloromethane, dimethyl sulfoxide, ethanol, N,N'-dimethylacetamide, dimethylformamide, N-methyl-2-pyrrolidone, or tert-butanol was added. Additional Anavex 2-73 was added to these vials as needed to ensure a mobile slurry was observed. The slurries were agitated for approximately 72 hours using an incubator shaker, cycling between ambient temperature (approximately 20-25 °C) and 40 °C (2 hours at each temperature). After approximately 72 hours of temperature cycling, the saturated solution was separated from the slurry using a 0.45 μm syringe filter. Approximately one-quarter of each filtrate was added with tert-butyl methyl ether to precipitate Form I, which was characterized by XRPD.

[0192] (ii) Example 2 Approximately 100 mg of Anavex 2-73 was weighed into a sample vial, to which 0.5 mL or 1 mL of 2-ethoxyethanol, 1-propanol, acetone, acetonitrile, dichloromethane, dimethyl sulfoxide, ethanol, N,N'-dimethylacetamide, dimethylformamide, N-methyl-2-pyrrolidone, or tert-butanol was added. Additional Anavex 2-73 was added to the vials as needed to ensure a mobile slurry was observed. The slurry was agitated for approximately 72 hours using an incubator shaker, with temperature cycling between ambient and 40°C (2 hours at each temperature). After approximately 72 hours of temperature cycling, the slurry was filtered through a 0.45 μm filter to isolate Form I precipitate, which was characterized by XRPD.

[0193] (iii) Example 3 Anavex 2-73 was diluted with I 100% ethanol containing approximately 2.5% volume / volume water (or less). The precipitate was slurried in PA and stirred for 20 hours at 70° C. The Form I precipitate was then isolated by filtration and characterized by XRPD.

[0194] (iv) Example 4 Approximately 100 mg of Anavex2-73 was heated to approximately 200° C. in a 5 mL beaker containing ice at atmospheric pressure. The residue was collected and characterized by XRPD.

[0195] Example 2. Preparation of Form II, Hydrochloride Salt Form II can be obtained by crystallization of Anavex 2-73 from an organic solvent:water mixture, such as isopropyl alcohol (IPA):water (90:10). It can also be obtained by air evaporation at ambient temperature from 1-butanol, chloroform, ethanol, or tert-butanol. An example of Form II preparation is as follows.

[0196] (i) Example 1 Approximately 100 mg of Anavex 2-73 was weighed into a sample vial, to which 0.5 mL or 1 mL of water, IPA:water (95:5 v / v), or IPA:water (97.5:2.5 v / v) was added. Additional Anavex 2-73 was added to the vial as needed to ensure a mobile slurry was observed. The slurry was agitated for approximately 72 hours using an incubator shaker, with temperature cycling between ambient temperature and 40°C (2 hours at each temperature). After approximately 72 hours of temperature cycling, the saturated solution was separated from the slurry using a 0.45 μm syringe filter. To approximately one-quarter of each filtrate, tert-butyl methyl ether, or THF in the case of water, was added to precipitate Form II, which was characterized by XRPD.

[0197] (ii) Example 2 Approximately 100 mg of Anavex 2-73 was weighed into a sample vial, to which 0.5 mL or 1 mL of IPA containing 2.5-10% v / v water was added. Additional Anavex 2-73 was added to these vials as needed to ensure a mobile slurry was observed. The slurry was agitated for approximately 72 hours using an incubator shaker, with temperature cycling between ambient and 40 °C (2 hours at each temperature). After approximately 72 hours of temperature cycling, the slurry was filtered through a 0.45 μm filter to isolate Form II precipitate, which was characterized by XRPD.

[0198] (iii) Example 3 Anavex 2-73 was slurried in IPA containing approximately 2.5-92.5% volume / volume water and stirred for 20 hours at a temperature of 20° C. The Form II precipitate was then isolated by filtration and characterized by XRPD.

[0199] (iv) Example 4 Anavex 2-73 was slurried in IPA containing approximately 7.5-45% volume / volume water and stirred for 20 hours at a temperature of 70° C. The Form II precipitate was then isolated by filtration and characterized by XRPD.

[0200] (v) Example 5 Form II was produced at a 250 mg scale using both IPA:water (95:5% v / v) and IPA:water (97.5:2.5% v / v). Approximately 250 mg of Anavex 2-73 was weighed into a 20 mL scintillation vial. To each vial, 2.5 mL of either IPA:water (95:5% v / v) or IPA:water (97.5:2.5% v / v) was added. The slurries were incubated for approximately 48 hours using an incubator shaker with temperature cycling between ambient and 40°C (2 hours at each temperature). The mixture was stirred for approximately 48 hours. After approximately 48 hours of temperature cycling, the saturated solution was separated from the slurry. The saturated solution was then allowed to evaporate at ambient temperature. The remaining solid material after temperature cycling was analyzed by XRPD. The remaining solid was air-dried at ambient temperature and again analyzed by XRPD.

[0201] Example 3. Preparation of Form III, Chloride Salt Form III can be obtained by crystallization of Anavex 2-73 from water and isopropyl alcohol (IPA) at 20°C.

[0202] In some embodiments, the preparation of Form III is as follows.

[0203] (i) Example 1 In one embodiment, an example of the preparation of Form III is as follows: Approximately 100 mg of Anavex 2-73 was weighed into a sample vial, and 0.5 mL of water was added thereto. Additional Anavex 2-73 was added to the vial to ensure a mobile slurry was observed. The slurry was agitated for approximately 72 hours using an incubator shaker with temperature cycling between ambient temperature and 40°C (2 hours at each temperature). After approximately 72 hours of temperature cycling, the saturated solution was separated from the slurry using a 0.45 μm syringe filter. Approximately one-quarter of the filtrate was evaporated at ambient temperature (approximately 20°C). XRPD data was collected on the solid residue.

[0204] (ii) Example 2 In another embodiment, an example of the preparation of Form III is as follows: Anavex2-73 was slurried in IPA and stirred for 20 hours at a temperature of 20° C. The Form III precipitate was then isolated by filtration and characterized by XRPD.

[0205] (iii) Example 3 In one embodiment, an example of the preparation of Form III is as follows: Form III material was produced on a 500 mg scale using water. Approximately 500 mg of Anavex 2-73 was weighed into a 20 mL scintillation vial and 600 μL of water was added. The slurry was then agitated using an incubator shaker with temperature cycling between ambient temperature and 40° C. (2 hours at each temperature) for approximately 48 hours. After approximately 24 hours, a thin slurry was observed, and an additional 130 mg of Anavex 2-73 was added to the slurry. After the full 48-hour temperature cycle, the saturated solution was separated from the slurry. The saturated solution was then allowed to evaporate at ambient temperature. The remaining solid material after temperature cycling was analyzed by XRPD. The remaining solid was air-dried at ambient temperature and reanalyzed by XRPD.

[0206] Example 4. Preparation of Form IV, Hydrochloride Salt Form IV was obtained by lyophilization of Anavex 2-73 from water. In one embodiment, the preparation of Form IV is as follows.

[0207] (i) Example 1 Approximately 20 mg of Anavex 2-73 was weighed into a 2 mL sample vial and dissolved in 200 μL of deionized water. The sample was then placed in a −20° C. freezer. Once frozen, the sample was lyophilized and characterized by XRPD to assess the crystalline nature of the material.

[0208] Example 5. Preparation of Form V, Hydrochloride Salt Form V was obtained by rotary evaporation of Anavex 2-73 from dichloromethane. In one embodiment, Form V is prepared as follows.

[0209] (i) Example 1 Approximately 20 mg of Anavex 2-73 was dissolved in 300 μL of dichloromethane and rapidly evaporated in a fume hood using a rotary evaporator. XRPD data was collected on the resulting solid material.

[0210] Example 6. Preparation of Form VI, Hydrochloride Salt Form VI was obtained after rapid cooling of an aqueous solution of Anavex 2-73 to 5° C. A specific example of the preparation of Form VI is as follows.

[0211] (i) Example 1 Approximately 100 mg of Anavex 2-73 was weighed into a sample vial, to which 0.5 mL of water was added. Additional Anavex 2-73 was added to the vial to ensure a mobile slurry was observed. The slurry was agitated for approximately 72 hours using an incubator shaker, with temperature cycling between ambient temperature and 40°C (2 hours at each temperature). After approximately 72 hours of temperature cycling, the saturated solution was separated from the slurry using a 0.45 μm syringe filter. Approximately one-quarter of the filtrate was placed in a refrigerator at 5°C and stored until a solid precipitate was observed. XRPD data was collected on the wet precipitate to prevent potential desolvation.

[0212] Example 7. Preparation of Form VII, Hydrochloride Salt Form VII was obtained by air evaporation of Anavex 2-73 from methanol. A specific example of the preparation of Form VII is as follows.

[0213] (i) Example 1 Approximately 100 mg of Anavex 2-73 was weighed into a sample vial, to which 0.5 mL of methanol was added. Additional Anavex 2-73 was added to the vial to ensure a mobile slurry was observed. The slurry was agitated for approximately 72 hours using an incubator shaker with temperature cycling between ambient temperature and 40°C (2 hours at each temperature). After approximately 72 hours of temperature cycling, the saturated solution was separated from the slurry using a 0.45 μm syringe filter. Approximately one-quarter of the filtrate was evaporated at ambient temperature (approximately 20°C). XRPD data was collected on the solid residue.

[0214] Example 8. Preparation of Form VIII, Hydrochloride Salt Form VIII is obtained by slurrying Anavex 2-73 in water at 20° C. A specific example of the preparation of Form VIII is as follows.

[0215] (i) Example 1 Anavex 2-73 was added to approximately 1.5 mL of water at 20° C. until a slurry was obtained, and the suspended material was analyzed using XRPD after slurried for 20 hours.

[0216] (ii) Example 2 Anavex 2-73 was completely dissolved in water and allowed to air evaporate at 20° C. until precipitation was observed. The resulting Form VIII was isolated while still wet with water and, upon drying, converted to Form I. XRPD patterns were collected before and after drying.

[0217] Example 9. Preparation of Form I, Free Base To a 250 mL separatory funnel was added 150 mL of EtOAc and 500 mg of A2-73 (hydrochloride salt), followed by 100 mL of concentrated NaHCO. The resulting mixture was shaken and the aqueous layer was removed. Two additional 100 mL aliquots of concentrated NaHCO were added, each time shaking the mixture and removing the aqueous layer.

[0218] The organic layer was washed with 100 mL of deionized water, then dried over magnesium sulfate, and then filtered. The filtrate was collected, and the EtOAc was removed using a rotary evaporator. After evaporation, a clear oil was obtained, which was dried under a stream of nitrogen to give a white solid. The resulting solid was weighed and analyzed by XRPD.

[0219] The data show that A2-73 free base is (i) crystalline, (ii) not highly water soluble (unlike the HCl salt), and (iii) non-hygroscopic. Its molecular weight (MW) of 280 is below the accepted general transdermal cutoff MW of approximately 400, and its calculated LogP is 3.5. In addition, because A2-73 free base has only two hydrogen bond donor / acceptor sites, it falls below the general rule-of-thumb limit of approximately five hydrogen bond donor / acceptor sites, which would potentially limit transdermal delivery.

[0220] Therefore, A2-73 free base is a useful active pharmaceutical ingredient in transdermal extended-release formulations. Reference is made to an oral dose of 20 mg of A2-73. Transdermal administration is performed twice weekly at a significantly lower dosage, based on the belief that avoiding first-pass liver or liver first-pass effect is a beneficial outcome of the present invention. The transdermal patch matrix layer may usefully contain, as non-limiting examples, oleyl oleate, povidone K90, levulinic acid, or cross-linked poly[acrylic acid-co-butyl acrylate-co-(2-ethylhexyl)acrylate-co-vinyl acetate]. It is also important to note the use of absorption enhancers (penetration enhancers) in the matrix formulation to generate a high flux of the active compound when the system is applied to the skin. Typical known enhancers include ethanol, glycerol monolaurate, DMF, polyethylen glycol monolaurate, etc.

[0221] Example 10. Preparation of Sulfate Salt Form I A2-73 sulfate Form I was obtained by addition of sulfuric acid (in THF) to A2-73 free base in ethanol, THF, acetone, 2-propanol, or 2-ethoxyethanol. A specific example of the preparation of A2-73 sulfate Form I is as follows.

[0222] (i) Example 1 Approximately 200 mg of A2-73 free base was weighed into a 20 mL scintillation vial, and then 4 mL of ethanol was added to the vial. The apparent pH of the resulting solution was determined using a pH meter. A sulfuric acid stock solution (1119.4 μL of a solution containing 4.1 μL of 98% sulfuric acid per 74.6 μL of THF) was added to the vial, followed by stirring. The apparent pH of the solution was measured again. The sample was then temperature cycled between ambient temperature and 40°C for approximately 24 hours. At the end of approximately 24 hours, no solids were observed, so the sample was left uncapped in a fume hood and allowed to air evaporate for approximately 72 hours, followed by drying under a nitrogen stream. Since no solids were observed, the sample was placed in a vacuum oven for 1 hour, yielding a colorless, sticky solid. The sample was further dried in the vacuum oven for approximately 4 hours, yielding a white solid, which was characterized by XRPD.

[0223] (ii) Example 2 A stock solution of sulfuric acid was prepared in THF (272.0 μL sulfuric acid in 4728.0 μL THF). Separately, 20 mg of A2-73 free base was weighed into 1.5 mL HPLC vials, and 300 μL of the appropriate solvent (THF, ethanol, acetone, 2-propanol, or 2-ethoxyethanol) was added to each vial along with 74.6 μL of the acid stock solution (1.05 equivalents of acid). Samples were temperature cycled between ambient and 40°C for 72 hours in 4-hour cycles. XRPD data was collected on the isolated precipitates.

[0224] Example 11. Preparation of Sulfate Salt Form II A2-73 sulfate Form II is prepared by dissolving A2-73 free salt in acetonitrile with sulfuric acid (in THF). A specific example of the preparation of A2-73 sulfate Form II is as follows:

[0225] (i) Example 1 A stock solution of sulfuric acid was prepared in THF (272.0 μL sulfuric acid in 4728.0 μL THF). Separately, 20 mg of A2-73 free base was weighed into a 1.5 mL HPLC vial, and 300 μL of acetonitrile was added along with 74.6 μL of the acid stock solution (1.05 equivalents of acid). The sample was temperature cycled between ambient and 40° C. for 72 hours in 4-hour cycles, and the resulting precipitate was analyzed by XRPD.

[0226] Example 12. Preparation of Mesylate Form I A2-73 mesylate Form I was obtained by addition of methanesulfonic acid (in THF) to A2-73 free base in ethanol, acetonitrile, acetone, 2-propanol, or 2-ethoxyethanol. A specific example of the preparation of A2-73 mesylate Form I is as follows.

[0227] (i) Example 1 A stock solution of methanesulfonic acid was prepared in THF (324.4 μL methanesulfonic acid in 4675.6 μL THF). Separately, 20 mg of A2-73 free base was weighed into a 1.5 mL HPLC vial, to which 300 μL of the appropriate solvent (ethanol, acetonitrile, acetone, 2-propanol, or 2-ethoxyethanol) was added along with 74.6 μL of the acid stock solution (1.05 equivalents of acid). The samples were temperature cycled between ambient and 40°C for 72 hours in 4-hour cycles. The samples were filtered, and approximately 100 μL of the mother liquor from each salt-forming reaction was added to a 2 mL glass vial. The vials were left uncapped in a fume hood to allow evaporation. The observed post-evaporation solids were analyzed by XRPD.

[0228] Example 13. Preparation of Oxalate Salt Form I A2-73 oxalate Form I was obtained by addition of oxalic acid (in THF) to A2-73 free base in THF. A specific example of the preparation of A2-73 oxalate Form I is as follows.

[0229] (i) Example 1 A stock solution of oxalic acid was prepared in THF (450.2 μL oxalic acid in 4549.8 μL THF). Separately, 20 mg of A2-73 free base was weighed into a 1.5 mL HPLC vial, to which 300 μL of THF was added along with 74.6 μL of the acid stock solution (1.05 equivalents of acid). The sample was temperature cycled between ambient and 40° C. in 4-hour cycles for 72 hours, and the resulting precipitate was analyzed by XRPD.

[0230] Example 14. Preparation of Oxalate Salt Form II A2-73 oxalate Form II was obtained by adding oxalic acid (in THF) to A2-73 free base in acetone. A specific example of the preparation of A2-73 oxalate Form II is as follows.

[0231] (i) Example 1 A stock solution of oxalic acid was prepared in THF (450.2 μL oxalic acid in 4549.8 μL THF). Separately, 20 mg of A2-73 free base was weighed into a 1.5 mL HPLC vial, to which 300 μL of acetone was added along with 74.6 μL of the acid stock solution (1.05 equivalents of acid). The sample was temperature cycled between ambient and 40° C. in 4-hour cycles for 72 hours, and the resulting precipitate was analyzed by XRPD.

[0232] Example 15. Preparation of Oxalate Salt Form III A2-73 oxalate Form III was obtained by adding oxalic acid (in THF) to A2-73 free base in ethanol. A specific example of the preparation of A2-73 oxalate Form III is as follows.

[0233] (i) Example 1 Approximately 200 mg of A2-73 free base was weighed into a 20 mL scintillation vial, and then 4 mL of ethanol was added to the vial. The apparent pH of the resulting solution was determined using a pH meter. An oxalic acid stock solution (1119.4 μL of a solution containing 6.72 mg of phosphoric acid per 74.6 μL of THF) was added to the vial (1.05 equivalents) followed by stirring. The apparent pH of the solution was again measured. The sample was then temperature cycled between ambient temperature and 40° C. for approximately 24 hours, and the precipitated solid was isolated at the end of the approximately 24 hour period, air-dried for approximately 72 hours, and subsequently characterized by XRPD.

[0234] (ii) Example 2 A stock solution of oxalic acid was prepared in THF (450.2 μL oxalic acid in 4549.8 μL THF). Separately, 20 mg of A2-73 free base was weighed into a 1.5 mL HPLC vial, to which 300 μL of ethanol was added along with 74.6 μL of the acid stock solution (1.05 equivalents of acid). The sample was temperature cycled between ambient and 40° C. for 72 hours in 4-hour cycles, and the resulting precipitate was analyzed by XRPD.

[0235] (iii) Example 3 A stock solution of oxalic acid was prepared in THF (450.2 μL oxalic acid in 4549.8 μL THF). Separately, 20 mg of A2-73 free base was weighed into a 1.5 mL HPLC vial, to which 300 μL of ethanol was added along with 74.6 μL of the acid stock solution (1.05 equivalents of acid). The sample was temperature cycled between ambient and 40°C for 72 hours in 4-hour cycles. The sample was filtered, and approximately 100 μL of the filtrate was added to a 2 mL glass vial. The vial was left uncapped in a fume hood to allow evaporation. The observed post-evaporation solid was analyzed by XRPD.

[0236] (iv) Example 4 A stock solution of oxalic acid was prepared in THF (450.2 μL oxalic acid in 4549.8 μL THF). Separately, 20 mg of A2-73 free base was weighed into a 1.5 mL HPLC vial, to which 300 μL of ethanol was added along with 74.6 μL of the acid stock solution (1.05 equivalents of acid). The sample was temperature cycled between ambient and 40°C in 4-hour cycles for 72 hours. The sample was filtered, and approximately 100 μL of the filtrate was weighed into a 1.5 mL HPLC vial. The vial was capped and placed in a refrigerator at approximately 5°C for approximately 24 hours. The sample was checked periodically, and any solids observed were analyzed by XRPD. If the sample remained in solution, it was placed in a freezer at approximately -20°C for approximately 24 hours. The sample was checked periodically, and any solids observed were analyzed by XRPD.

[0237] (v) Example 5 A stock solution of oxalic acid was prepared in THF (450.2 μL oxalic acid in 4549.8 μL THF). Separately, 20 mg of A2-73 free base was weighed into a 1.5 mL HPLC vial, to which 300 μL of ethanol was added along with 74.6 μL of the acid stock solution (1.05 equivalents of acid). The sample was temperature cycled between ambient and 40°C for 72 hours in 4-hour cycles. The sample was filtered, and approximately 100 μL of the filtrate was transferred to a 1.5 mL HPLC vial. 100 μL aliquots of tert-methyl ether were added until precipitation was observed. XRPD data was collected on the precipitate.

[0238] Example 16. Preparation of Dihydrogen Phosphate Form I A2-73 dihydrogen phosphate Form I was obtained by addition of phosphoric acid (in THF) to a solution of A2-73 free base in THF, ethanol, acetonitrile, acetone, 2-propanol, or 2-ethoxyethanol.

[0239] A specific example of the preparation of A2-73 dihydrogen phosphate Form I is as follows.

[0240] (i) Example 1 Approximately 300 mg of A2-73 free base was weighed into a 20 mL scintillation vial, and then 4 mL of acetone was added to the vial. The apparent pH of the resulting solution was determined using a pH meter. Phosphate stock solution (1119.4 μL of solution containing 7.31 mg of phosphoric acid per 74.6 μL of acetone) was added to the vial (1.05 equivalents), followed by stirring. The apparent pH of the solution was measured again. The sample was then temperature cycled between ambient temperature and 40° C. for approximately 24 hours, and the precipitated solid was isolated at the end of approximately 24 hours, air-dried for approximately 72 hours, and subsequently characterized by XRPD.

[0241] (ii) Example 2 A stock solution of phosphoric acid was prepared in THF (490 μL of phosphoric acid in 4510 μL of THF). Separately, 20 mg of A2-73 free base was weighed into a 1.5 mL HPLC vial, and 300 μL of the appropriate solvent (THF, ethanol, acetonitrile, acetone, 2-propanol, or 2-ethoxyethanol) was added along with 74.6 μL of the acid stock solution (1.05 equivalents of acid). The sample was temperature cycled between ambient and 40°C for 72 hours in 4-hour cycles, and the resulting precipitate was analyzed by XRPD.

[0242] Example 17. Preparation of Edisylate Salt Form I A2-73 edisylate Form I was obtained by adding 1,2-ethanedisulfonic acid (in THF) to A2-73 free base in ethanol, acetonitrile, acetone, 2-propanol, or 2-ethoxyethanol. A specific example of the preparation of A2-73 edisylate Form I is as follows.

[0243] (i) Example 1 A stock solution of 1,2-ethanedisulfonic acid was prepared in THF (1170.7 μL of 1,2-ethanedisulfonic acid in 3829.3 μL of THF). Separately, 20 mg of A2-73 free base was weighed into a 1.5 mL HPLC vial, to which 300 μL of the appropriate solvent (ethanol, acetonitrile, acetone, 2-propanol, or 2-ethoxyethanol) was added along with 74.6 μL of the acid stock solution (1.05 equivalents of acid). The samples were temperature cycled between ambient and 40°C for 72 hours in 4-hour cycles. The samples were filtered, and approximately 100 μL of each filtrate was added to a 2 mL glass vial. The vials were left uncapped in a fume hood to allow evaporation. The observed post-evaporation solids were analyzed by XRPD.

[0244] (ii) Example 2 A stock solution of 1,2-ethanedisulfonic acid was prepared in THF (1170.7 μL of 1,2-ethanedisulfonic acid in 3829.3 μL of THF). Separately, 20 mg of A2-73 free base was weighed into a 1.5 mL HPLC vial, to which 300 μL of the appropriate solvent (ethanol, acetone, or 2-propanol) was added along with 74.6 μL of the acid stock solution (1.05 equivalents of acid). The samples were temperature cycled between ambient and 40°C for 72 hours in 4-hour cycles. The samples were filtered, and approximately 100 μL of each filtrate was weighed into a 1.5 mL HPLC vial. The vials were capped and refrigerated at approximately 5°C for approximately 24 hours. The sample was placed in a freezer. The sample was checked periodically and any solids observed were analyzed by XRPD. The sample, which appeared as a solution, was placed in a freezer at approximately -20°C for approximately 24 hours. The sample was checked periodically and any solids observed were analyzed by XRPD.

[0245] Example 18. Preparation of Benzoate Salt Form I A2-73 benzoate Form I was obtained by addition of benzoic acid (in THF) to A2-73 free base in THF, acetonitrile, or acetone. A specific example of the preparation of A2-73 benzoate Form I is as follows.

[0246] (i) Example 1 A stock solution of benzoic acid was prepared in THF (610.6 μL benzoic acid in 4389.4 μL THF). Separately, 20 mg of A2-73 free base was weighed into a 1.5 mL HPLC vial, to which 300 μL of the appropriate solvent (THF, acetonitrile, or acetone) was added along with 74.6 μL of the acid stock solution (1.05 equivalents of acid). The samples were temperature cycled between ambient and 40°C for 72 hours in 4-hour cycles. The samples were filtered, and approximately 100 μL of each filtrate was added to a 2 mL glass vial. The vials were left uncapped in a fume hood to allow evaporation. The observed post-evaporation solids were analyzed by XRPD.

[0247] Example 19. Preparation of hydrogen fumarate form I A2-73 fumarate salt Form I was obtained by addition of fumaric acid (in THF) to A2-73 free base in ethanol or THF. A specific example of the preparation of A2-73 fumarate salt Form I is as follows.

[0248] (i) Example 1 Approximately 200 mg of A2-73 free base was weighed into a 20 mL scintillation vial, and then 4 mL of ethanol was added to the vial. The apparent pH of the resulting solution was determined using a pH meter. Fumaric acid stock solution (1119.4 μL of solution containing 8.66 mg of fumaric acid per 74.6 μL of ethanol) was added to the vial (1.05 equivalents), followed by stirring. The apparent pH of the solution was measured again. The sample was then temperature cycled between ambient temperature and 40° C. for approximately 24 hours. Since no solids were observed at the end of approximately 24 hours, the sample was left uncapped in a fume hood and allowed to air evaporate for approximately 72 hours, followed by characterization by XRPD.

[0249] A stock solution of fumaric acid was prepared in THF (580.3 μL fumaric acid in 4419.7 μL THF). Separately, 20 mg of A2-73 free base was weighed into a 1.5 mL HPLC vial, to which 300 μL of THF was added along with 74.6 μL of the acid stock solution (1.05 equivalents of acid). The sample was temperature cycled between ambient and 40°C for 72 hours in 4-hour cycles. The sample was filtered, and approximately 100 μL was transferred to a 2 mL glass vial. The vial was left uncapped in a fume hood to allow evaporation. The observed post-evaporation solid was analyzed by XRPD.

[0250] A stock solution of fumaric acid was prepared in THF (580.3 μL fumaric acid in 4419.7 μL THF). Separately, 20 mg of A2-73 free base was weighed into a 1.5 mL HPLC vial, to which 300 μL of THF was added along with 74.6 μL of the acid stock solution (1.05 equivalents of acid). The sample was temperature cycled between ambient temperature and 40°C for 72 hours in 4-hour cycles. The sample was filtered, and approximately 100 μL of the filtrate was transferred to a 1.5 mL HPLC vial. 100 μL aliquots of tert-butyl methyl ether were added until precipitation was observed, and the resulting precipitate was analyzed by XRPD.

[0251] Example 20 Preparation of Fumarate Form II A2-73 fumarate Form II was obtained by adding fumaric acid (in THF) to A2-73 free base in ethanol. A specific example of the preparation of A2-73 fumarate Form II is as follows.

[0252] (i) Example 1 A stock solution of fumaric acid was prepared in THF (580.3 μL fumaric acid in 4419.7 μL THF). Separately, 20 mg of A2-73 free base was weighed into a 1.5 mL HPLC vial, to which 300 μL of ethanol was added along with 74.6 μL of the acid stock solution (1.05 equivalents of acid). The sample was temperature cycled between ambient and 40° C. in 4-hour cycles for 72 hours, and the resulting precipitate was analyzed by XRPD.

[0253] Example 21. Preparation of Fumaric Acid Hydrogen Form III A2-73 hydrogen fumarate Form III was obtained by addition of fumaric acid (in THF) to A2-73 free base in IPA. A specific example of the preparation of A2-73 hydrogen fumarate Form III is as follows:

[0254] (i) Example 1 Approximately 300 mg of A2-73 free base was weighed into a 20 mL scintillation vial, and then 4 mL of IPA was added to the vial. The apparent pH of the resulting solution was determined using a pH meter. Fumaric acid stock solution (1119.4 μL of a 1 M fumaric acid stock solution in IPA) was added to the vial (1.05 equivalents), followed by stirring. The apparent pH of the solution was measured again. The sample was then temperature cycled between ambient temperature and 40° C. for approximately 24 hours, and the precipitated solid was isolated by centrifugation at the end of approximately 24 hours. The isolated solid was air-dried for approximately 72 hours and subsequently characterized by XRPD.

[0255] A stock solution of fumaric acid was prepared in THF (580.3 μL fumaric acid in 4419.7 μL THF). Separately, 20 mg of A2-73 free base was weighed into a 1.5 mL HPLC vial, to which 300 μL of IPA was added along with 74.6 μL of the acid stock solution (1.05 equivalents of acid). The sample was temperature cycled between ambient and 40° C. in 4-hour cycles for 72 hours, and the resulting precipitate was analyzed by XRPD.

[0256] Example 22. Preparation of Fumarate Salt Form IV A2-73 fumarate salt Form IV was obtained by addition of fumaric acid (in THF) to A2-73 free base in 2-ethoxyethanol. A specific example of the preparation of A2-73 fumarate salt Form IV is as follows.

[0257] (i) Example 1 A stock solution of fumaric acid was prepared in THF (580.3 μL fumaric acid in 4419.7 μL THF). Separately, 20 mg of A2-73 free base was weighed into a 1.5 mL HPLC vial, to which 300 μL of 2-ethoxyethanol was added along with 74.6 μL of the acid stock solution (1.05 equivalents of acid). The sample was temperature cycled between ambient and 40°C for 72 hours in 4-hour cycles. The sample was filtered, and approximately 100 μL was transferred to a 2 mL glass vial. The vial was left uncapped in a fume hood to allow evaporation. The observed post-evaporation solid was analyzed by XRPD.

[0258] Example 23. Anavex 2-73 fumarate form V A2-73 fumarate salt Form V was obtained by addition of fumaric acid (in THF) to A2-73 free base in IPA. A specific example of the preparation of A2-73 fumarate salt Form V is as follows.

[0259] (i) Example 1 Approximately 300 mg of A2-73 free base was weighed into a 20 mL scintillation vial, and then 4 mL of IPA was added to the vial. The apparent pH of the resulting solution was determined using a pH meter. Fumaric acid stock solution (1119.4 μL of a 1 M fumaric acid stock solution in IPA) was added to the vial (1.05 equivalents), followed by stirring. The apparent pH of the solution was measured again. The sample was then temperature cycled between ambient temperature and 40° C. for approximately 24 hours, and the precipitated solid was isolated by centrifugation at the end of approximately 24 hours. The isolated solid was air-dried for approximately 72 hours and subsequently characterized by XRPD.

[0260] Example 24. Transdermal patch A 63-year-old man presents with early signs of Alzheimer's disease. He is administered a pharmaceutical composition via a transdermal patch containing 100 mg of ANAVEX 2-73 free base, replacing a 4 cm patch approximately every 3 days for 120 days. Cognitive function remains stable over that time, with no further cognitive loss detected.

[0261] Example 25. Transdermal patch A 57-year-old woman presents with early signs of Alzheimer's disease. Anavex 2-73 is administered via a 9cm transdermal patch containing 200mg of a pharmaceutical composition containing free base, with the patch replaced weekly for 180 days. A blood level of approximately 12ng / ml is maintained. Cognitive function remains stable throughout this time, with no further cognitive loss detected.

[0262] Example 26. Extended Release Oral Dosage Form An 84-year-old man with unspecified progressive dementia received 30 mg of ANAVEX 2-73 fumarate in enteric-coated tablets every other day for 180 days. Blood levels revealed approximately 25 mg per day of administration. Cognitive function remained stable over that time, with no additional cognitive loss detected.

[0263] Example 27. Extended Release Oral Dosage Form A 77-year-old woman with unspecified progressive dementia received 50 mg of ANAVEX 2-73 free base in enteric-coated tablets every other day for 180 days. Blood levels revealed approximately 20 mg per day of administration. Cognitive function remained stable over that time, with no additional cognitive loss detected.

[0264] Example 28. The Sig1-R agonist ANAVEX2-73 enhances autophagy activity. To study the effect of ANAVEX2-73 on autophagy, human HeLa cells were treated with the compound and autophagic activity was analyzed by examining the flux of LC3-II. LC3-II is a lipidated form of LC3 that remains (partially) bound to autophagosomes and is thus degraded by lysosomes. Therefore, quantification of LC3-II flux using bafilomycin A1 (BafiA1, 2 μM) to inhibit lysosomal degradation directly corresponds to cellular autophagic activity. As shown in Figure 35, ANAVEX2-73 significantly induced autophagic flux compared to control conditions. There was a concentration-dependent and significant increase in autophagic flux after application of ANAVEX2-73: a more than 2-fold increase at 10 μM and a more than 1.5-fold increase at 1 μM ANAVEX2-73 (Figure 35A). As a standard positive control to trigger the induction of autophagy, HeLa cells were incubated with EBSS, which resembles nutrient deprivation as an autophagy stimulus.

[0265] ANAVEX2-73 and other known experimental Sig-1R agonists were used in the experiments. Such compounds include (+)-pentazocine, (+)-SKF10,047, SA4503 (14243,4-dimethoxyphenyldecyl)-4-(3-phenylpropionyl)-4-(2-methyl-4-phenylpropionyl)-4-(3-phenylpropionyl)-4-(2-methyl ... These include anabexil (piperazine), and PRE-084 (2-morpholin-4-ylethyl 1-phenylcyclohexane-1-carboxylate). In contrast to ANAVEX2-73, PRE-084 and other experimental compounds are not applicable to clinical studies for various reasons. However, because the Sig-1R ligand PRE-084 exhibits activity in the central nervous system in animal models, such as nootropic and antidepressant activity, this compound was included in some of the flux assays as a control. PRE-084 was also found to promote autophagic flux in HeLa cells (Figure 35B). At 1 μM, PRE-084 induced autophagic flux greater than 1.5-fold, comparable to that of ANAVEX2-73 at the same concentration (Figure 35B).

[0266] Next, we complemented the Western blot experiments by directly visualizing the extent of autophagosome appearance in HEK293 cells. To do so, we applied ANAVEX2-73 (1 µM) to HEK293 cells stably expressing a GFP-LC3B reporter construct. This cellular model allows us to directly monitor the accumulation of LC3-II-positive autophagosome structures upon BafiA1 supplementation by confocal fluorescence microscopy. Indeed, ANAVEX2-73 treatment resulted in an overall increased number of LC3-II-positive puncta and autophagic flux (Figure 35C).

[0267] In summary, Sig-1R activation induced significantly increased autophagic flux in both independent cellular assays and two different human cell lines. Part of the effect of ANAVEX2-73 as a Sig-1R ligand could potentially be attributed to its effect at muscarinic ACh receptors. However, little is known about the influence of mACh receptors on autophagy. In fact, to date, only one report in the literature has demonstrated that ACh-induced autophagy has a cytoprotective effect through muscarinic ACh receptor-activated AMPK-mTOR pathway. On the other hand, our finding that PRE-084, a unique selective Sig-1R agonist, also induced autophagic flux strongly supports the effect of ANAVEX2-73 on autophagy as mediated by Sig-1R activation. Furthermore, as shown in Example 30 below, there is no experimental data demonstrating that activation of muscarinic ACh receptors has beneficial effects on protein aggregation and proteostasis, as clearly demonstrated by ANAVEX2-73.

[0268] Example 29. Sig-1R activation induces ULK1 phosphorylation and affects the expression levels of different autophagy network factors. Activation of the serine / threonine protein kinase ULK1 (unc-51-like kinase 1) via phosphorylation at serine 555 indicates stimulation of the canonical autophagy pathway. ANAVEX2-73 significantly induced ULK1 serine 555 phosphorylation (up to 2-fold at 1 μM, Figure 36A). PRE-084 was also analyzed as a Sig-1R agonist and was found to similarly promote ULK1 serine 555 phosphorylation (up to 1.5-fold at 1 μM, Figure 36B). It should be noted that this activated ULK1 phosphorylation can be inhibited not only by mTOR but also stimulated via AMPK kinase. Both are fundamental physiological sensors of nutritional status and important signal transducers for canonical autophagy stimulation. ULK1 is, in fact, the signal mediating the induction of phagocyte formation during the autophagic process and, therefore, a central promoter of autophagy. ULK1 itself functions in a complex with at least three protein partners: FIP200 (200 kDa focal adhesion kinase family interacting protein), ATG (autophagy-related protein) 13 (ATG13), and ATG101. The fact that the complex patterns of upstream pathways (including mTOR and AMPK) converge on ULK1 indicates that this complex functions as a node, transducing multiple signals into autophagosome formation.

[0269] Sig-1R activation markedly induces ULK1 phosphorylation and autophagic flux Given our findings, we next investigated the relative expression levels of key autophagy network factors, which represent distinct set points in the autophagic process, after treatment of HeLa cells with ANAVEX2-73 using a PCR autophagy array (Figure 36C). Most notably, we found ANAVEX2-73-mediated induction of mRNA expression of GABA type A receptor-associated protein-like 1 (GABARAPL1, expression level approximately 2.7; the cutoff for induction was set at an expression level of 1.5), which, like GABARAP, is associated with autophagic vesicles and is involved in the autophagic process.

[0270] GABARAPL1 belongs to the human MAP1LC3 family, which consists of six ATG8 orthologs, MAP1LC3A, MAP1LC3B, and MAP1LC3C, and three MAP1LC3 paralogs, the GABA receptor-associated protein GABARAP1, GABARAPL1, and GABARAPL2, which have partially redundant roles in autophagy. Additionally, the expression of the ubiquitin and autophagy receptor SQSTM1 / p62, involved in the selective macroautophagy pathway, was enhanced by ANAVEX2-73 (expression levels of approximately 2.9). Furthermore, there was a clear trend toward the induction of ATG12, which is conjugated to ATG5 and assembles the autophagosomal protein complex that ultimately acts together with ATG16L1 in autophagosome biogenesis. Consistently, ATG16L1 expression also appeared to be enhanced after treatment of cells with ANAVEX2-73 (Figure 36C). Furthermore, it is clear that none of the autophagy network factors included in this PCR array was downregulated in its expression upon treatment with ANAVEX2-27, supporting the important finding that Sig-1R activation has a positive regulatory effect on autophagy.

[0271] Example 30. ANAVEX2-73 positively regulates autophagy, increases proteostasis capacity, and ameliorates protein aggregation-mediated paralysis in C. elegans. The in vitro autophagy regulation by ANAVEX2-73 and its effects on several key autophagy network components prompted us to further analyze the effects of ANAVEX2-73-mediated Sig-1R activation on autophagy and proteostasis in vivo using a C. elegans model. The C. elegans orthologue of human Sig-1R is W08F4.3, which is expressed in several tissues, including the musculature. To monitor autophagic flux in vivo, we used a GFP-LGG-1 reporter worm strain. LGG-1 is the C. elegans orthologue of mammalian GABARAP, allowing us to assess autophagic activity using Western blotting and confocal fluorescence microscopy. Western blotting was used to analyze the levels of GFP-LGG-1-II with and without BafiA1, as well as flux measurements in HeLa cells, as shown in Figure 37. Indeed, ANAVEX2-73 (80 μM) significantly enhanced autophagic flux by nearly two-fold in C. elegans (worms treated with BafiA1 or DMSO for 6 hours; FIG. 37A).

[0272] To further substantiate this finding, we directly visualized autophagic structures, as indicated by GFP-LGG-1 positive puncta, using confocal fluorescence microscopy. ANAVEX2-73 supplementation (plus / minus BafiA1) significantly increased the number of GFP-LGG1 puncta, indicating increased autophagic activity. Treatment of worms with ANAVEX2-73 resulted in a relative increase in the number of puncta after BafiA1 treatment compared to control worms. Indeed, a significant increase was found; autophagic flux, as observed in vivo, was induced approximately 2.5-fold by ANAVEX2-73 (Figure 37B; the number of GFP-positive autophagosome structures (indicated by arrows) was counted in three independent experiments and in each experiment in the head region of at least 8–11 worms), which is consistent with Western blot analysis (Figure 37A).

[0273] Taken together, our in vitro and in vivo data clearly demonstrate that the Sig-1R agonist ANAVEX2-73 induces autophagy, as demonstrated by autophagic flux measurements. This encouraged us to further investigate the functional consequences of autophagy induction, focusing on the impact of degradation pathways on proteostasis in vivo. Therefore, we used human Aβ42-expressing worms, which are characterized by time-dependent paralysis due to the accumulation of Aβ42 oligomers and high-molecular-weight aggregates in body wall muscle cells. Here, we emphasize that Aβ42-expressing worms should not be considered as a model of AD, but rather as an experimental model of general proteostatic stress and proteotoxicity, in which protein aggregation in muscle cells leads to a distinct phenotype (here, paralysis). Aβ42 protein aggregates were stained in situ with thioflavin. Compared to control worms, treatment of Aβ42-worms with ANAVEX2-73 reduced the number of thioflavin-positive Aβ42 aggregates (Figure 38A; worms were treated with 80 μM ANAVEX2-73 or M9 medium (control) for 9 consecutive days), indicating that autophagy induction affects proteostasis and reduces tissue deposition of aggregates, possibly through enhanced clearance of Aβ42 aggregates. Accumulation of Aβ42 aggregates in muscle cells is known to result in enhanced paralysis of worms over time. To analyze the effect of ANAVEX2-73-induced autophagy on time-dependent locomotor behavior, we investigated the extent of this paralysis. C. elegans were treated with the compounds (or M9 buffer as a control) for up to 12 days, and paralysis was quantified daily. Using two concentrations of ANAVEX2-73 (50 and 100 μM), we found a clear reduction in paralysis in the two ANAVEX2-73-treated groups, which clearly distinguished these groups from controls in terms of the degree of paralysis (Figure 38B; worms were maintained in the presence of ANAVEX2-73 or M9 buffer and examined daily for the paralysis phenotype). The paralysis fraction was significantly different comparing ANAVEX2-73-treated and control worms. Thus, ANAVEX2-73 clearly slowed the rate of paralysis in Aβ42-expressing worms, counteracting the time-dependent motor impairment.

[0274] The findings described herein that Sig-1R agonist-mediated autophagy induction directly affects proteostasis by reducing protein aggregation and proteotoxicity-induced behavioral impairment in worms indicate a role for Sig-1R activation in the prevention and treatment of neurodegeneration associated with imbalanced protein homeostasis. Consistent with the ANAVEX2-73-induced increase in proteostatic capacity observed herein, the involvement of Sig-1R deficiency or dysfunction has been described in ALS, a disorder characterized by severely disrupted protein homeostasis and characteristic intracellular protein aggregation. For example, (1) Sig-1R missense mutations can cause ALS, (2) knockout of Sig-1R accelerates disease in SOD1 mutant mice, and (3) ALS-associated mutant Sig-1R causes the accumulation of autophagic material and reduced autophagy. Furthermore, supporting the protective role of Sig-1R activity, (1) treatment with the experimental drug PRE-084 ameliorates SOD1 mouse pathology, (2) mutant Sig-1R expression induces cytoplasmic accumulation of ALS-associated TDP43 and FUS in cells, and (3) PRE-084 improves motor function and motor neuron survival in ALS mice. In complete agreement with the findings herein, overexpression of the Sig-1A receptor increases the number of p62 / SQSTM1 and LC3B puncta, indicative of autophagy activation, in human disease tissue.

[0275] Several steps in the autophagy process are amenable to therapeutic modulation, and different autophagy-activating compounds have already been studied at various experimental levels (in vitro and in vivo) and models of human diseases, including cancer and neurodegeneration. Regarding effective intervention of neurodegenerative disorders, of course, for any compound to be studied in humans in the context of the central nervous system, in addition to toxicity and safety issues, blood-brain barrier permeability must also be ensured. One example of a compound that targets autophagy is lithium, which has been used to treat bipolar disorder and acts as an activator of autophagy by interfering with upstream steps in autophagy induction. Metformin and simvastatin have also been shown experimentally to promote autophagy, possibly via activation of AMPK, and are used to treat diabetes and obesity, respectively. Sig-1R agonists are under intense investigation for the treatment of different neurodegenerative diseases, including AD and ALS. Without being bound by theory, it is the combination of receptor activities that may make ANAVEX2-73 an interesting compound for AD therapy.

[0276] Taken together, the results presented in Examples 28-30 herein demonstrate that Sig-1R activation (a) enhances autophagic flux in human cells and C. elegans and (b) has a positive effect on proteostasis. The novel activity of ANAVEX2-73, a compound with dual selective Sig-1R / muscarinic activity in neurons, is described. This drug's activity includes potent induction of autophagy in vitro and in vivo, resulting in increased proteostatic capacity and even beneficial effects on the time-dependent paralysis phenotype in Aβ42-expressing C. elegans. The specific induction of the autophagic process and subsequent stabilization of proteostasis in neurons represents a key step toward stabilizing neuronal survival and function and may help prevent age-related neurodegeneration.

[0277] Introduction of Examples 28-30 The pathogenesis of neurodegenerative disorders, including Alzheimer's disease and Parkinson's disease (AD, PD), as well as amyotrophic lateral sclerosis (ALS), is associated with disrupted protein homeostasis. Therefore, the control and maintenance of proteome integrity and proteostasis are of paramount importance. Cellular proteostasis involves protein folding, protein assembly, refolding of damaged proteins, and protein degradation, and is under the control of a fine-tuned network of factors, including chaperones such as heat shock protein 70 (HSP70) and distinct co-chaperones. For intact cell function and long-term survival, it is crucial to remove misfolded proteins through specialized processes. Two major cellular degradation pathways are the ubiquitin-proteasome system (UPS) and autophagy. The UPS is particularly important for physiological protein turnover but is limited in degradative substrates, while the autophagy-lysosomal pathway is responsible for the clearance of aggregated and disease-related proteins, particularly under pathogenic and aging conditions.

[0278] Autophagy is a highly dynamic, vesicle-mediated cellular degradation pathway involving double-membrane vesicles called autophagosomes that sequester large protein complexes (protein aggregates) and even entire organelles and deliver them to lysosomes for degradation. Under low nutrient and energy conditions, autophagy ensures energy supply by generating amino acid building blocks through recycling. In addition, autophagy plays an important role as a stress and adaptive response and rescue mechanism for maintaining cell survival and function. Canonical autophagy responds to environmental cues through various factors, primarily homologs of the autophagy-related (atg) genes originally identified in yeast. The mammalian target of rapamycin (mTOR) complex 1 (mTORC1) negatively regulates autophagic activity through the inhibitory phosphorylation of ULK1, a major early regulator of canonical autophagy. More downstream membrane swelling is regulated by two ubiquitin-like conjugation systems (ATG12-ATG5 and ATG8 / LC3) and the ATG18 protein family members of the WD repeat domain phosphoinositide interacting 1-3 (WIPI1-3).

[0279] There is a large body of data linking autophagy dysfunction and failure to neurodegenerative diseases, consistent with its role in proteostasis, leading to the accumulation of protein aggregates. Thus, modulation of autophagy has become one major pharmacological target in neurodegeneration. Indeed, there are multiple overlaps in autophagy and pathogenic pathways between AD, PD, and ALS. Recently, , Different alternative views and new pharmacological targets towards AD prevention and treatment are evolving, including a strong focus on the autophagic process.

[0280] There are two subtypes of Sigma receptors, Sigma-1 and Sigma-2, both of which are highly expressed in the central nervous system. The Sigma-1 receptor (Sig-1R) was cloned in 1996 and represents a 223-amino acid integral membrane protein localized to the endoplasmic reticulum (ER) (and the ER-mitochondria interface), suggesting its role as an ER chaperone. Sig-1R has been shown to promote cell survival by (1) ensuring Ca2+ signaling from the ER to mitochondria, (2) enhancing ER signaling to the nucleus, and (3) attenuating free radical damage by regulating the activity of the redox-responsive transcription factor Nrf2. Structurally, Sig-1R ligand binding has been characterized, and the crystal structure of the human receptor has been elucidated.

[0281] In general, deficiencies in Sig-1R expression or activity are associated with neurodegeneration, and activation of Sig-1R is associated with neuroprotection in different in vitro and in vivo models using different types of pharmacological Sig-1R activators with distinct pharmacological profiles. Pharmacological activation of Sig-1R leads to pluripotent regulatory downstream effects, and it has been strongly suggested that misfunction of Sig-1R is also involved in the pathogenesis of neurodegeneration. This is the basis for efforts to design novel and highly specific pharmacological Sig-1R activators for the therapy of neurodegenerative diseases, including:

[0282] In this context, a novel Sig-1R agonist, tetrahydro-N,N-dimethyl-1-2,2-diphenyl-1-3-furanmethanamine hydrochloride (ANAVEX 2-73), was developed. Pharmacologically, ANAVEX 2-73 exhibits mixed activity at Sig-1R and muscarinic receptors, acting with described affinities in the low micromolar range. Previous preclinical studies in animal models demonstrated the robust disease-modifying activity of ANAVEX 2-73. For AD, ANAVEX 2-73 was tested in a Phase 2a study in patients, demonstrating a favorable safety profile and concentration-dependent improvement on exploratory endpoints. Various neuromodulatory and neuroprotective effects, including mitochondrial protection, modulation of ERK activation, and promotion of astrocyte survival in mouse models of AD, as well as protection against oxidative stress, are also known for ANAVEX 2-73.

[0283] The first evidence of a possible link between Sig-1R, autophagy, and neurodegeneration was recently shown in the context of ALS. It was discovered that ALS-binding mutant Sig-1R caused the accumulation of autophagic material and actually reduced autophagy. In addition, small molecule Sig-1R modulators were found to induce autophagic degradation of programmed death-ligand 1 (PD-L1) in cancer cells. These findings prompted us to investigate the potential of ANAVEX2-73 to induce autophagy in human HeLa and HEK293 cells (in vitro) and C. elegans (in vivo) using standard measures for analyzing autophagic activity that have been well established by the present inventors. Furthermore, we investigated the effect of ANAVEX2-73 on protein aggregation and the subsequent impact of protein aggregates on motility behavior in C. elegans. Interestingly, ANAVEX2-73 is a potent inducer of autophagic flux in vitro and in vivo, ameliorating protein aggregate formation and paralysis in C. elegans.

[0284] Materials and Methods for Examples 28-30 Cell culture and microscopy. HeLa and HEK293A cells were cultured in DM supplemented with activated FBS (Life Technologies GmbH, Carlsbad, CA, USA, 10270106), 1x ABAM (Invitrogen, 15240-062), and 1 mM sodium pyruvate (Invitrogen, 1136-088). Cells were cultured in EM (Invitrogen, Carlsbad, USA, 41965062). After medium replacement, cells were treated with 10, 1, and 0.1 μM ANAVEX2-73 and PRE-084 (Tocris, Bristol, UK, 0589), respectively, for 2 h. ANAVEX2-73 was provided by ANAVEX Life Sciences Corp, New York, NY, USA. Bafilomycin A1 (Bafi.A1, 2 μM) (Toronto Research Chemicals, North York, ON, Canada, B110000) or DMSO was then added for an additional 2 h, and cells were finally harvested. Western blot analysis was performed as previously described [40, 41]. Briefly, cells were subjected to SDS-PAGE using precast NuPAGE 4%–12% Bis-Tris gels (Invitrogen, NP0322). Proteins were detected by chemiluminescence using an Amersham Imager 600 (GE).

[0285] Confocal fluorescence microscopy analysis of HEK293A cells stably expressing GFP-LC3B was performed with a laser scanning microscope LSM 710 (Zeiss, Oberkochen).

[0286] C. elegans strains, maintenance, and methods. C. elegans were maintained according to standard procedures on nematode growth medium (NGM) plates seeded with HB101 E. coli. In this study, the following strains were used: GFP::LGG-1 (ex[Plggl::Iggl::GFP] / pRF4), maintained at 20°C, and strain CL2006 (dvls2[pCL12(unc-54 / human Aβ peptide 1-42)+pRF4]), maintained at 15°C.

[0287] For analysis of paralysis rate, synchronized CL2006 nematodes were cultured at 15°C on plates seeded with HB101 E. coli resuspended in M9 (control) or 100 μM and 50 μM ANAVEX2-73. Starting on the first day of adulthood, worms were transferred to fresh plates daily and tested for paralysis by tapping their snouts with a platinum wire. Worms that moved their snouts but failed to move their bodies were scored as paralyzed. Dead worms or those displaying other phenotypes were not included in the statistics. Staining for amyloid-β42 aggregates using thioflavin S (Sigma T1892) was performed as previously described. Worms were mounted on 2% agar pads on glass slides, and confocal fluorescence microscopy analysis was performed with an LSM 710 (Zeiss, Oberkochen) laser scanning microscope.

[0288] For analysis of autophagic activity, synchronized nematodes expressing GFP::LGG-1 were cultured at 20 °C. On the first day of adulthood, worms were transferred to 80 µM ANAVEX2-73 or control M9 liquid culture medium for 2 h, followed by treatment with bafilomycin A1 or DMSO (control) for 4–6 h. Worms were then lysed for Western blot or analyzed by confocal fluorescence microscopy.

[0289] Western blot analysis was performed as previously described. Typically, 12 worms were subjected to SDS-PAGE using precast NuPAGE 4-12% Bis-Tris gels (Invitrogen, NP0322). Proteins were detected by chemiluminescence using a Fuji LAS-3000 dark box (Fujifilm, Dusseldorf). Aspects of the present invention include the following. [Appendix 1] A crystalline form of tetrahydro-N,N-dimethyl-2,2-diphenyl-3-furanmethanamine (A2-73), which is a salt or free base. [Appendix 2] 2. The crystalline form of claim 1, wherein the crystalline form is a pharmaceutically acceptable salt. [Appendix 3] 3. The crystalline form of claim 2, wherein the salt is selected from hydrochloride, fumarate, sulfate, dihydrogen phosphate, benzoate, mesylate, edysilate, and oxalate. [Appendix 4] 4. The crystalline form of claim 3, wherein the salt is a hydrochloride salt. [Appendix 5] Attachment 4. The crystalline form of claim 4, wherein the hydrochloride salt is characterized by the XRPD patterns shown in Figures 4, 6, 8, 9, 10, 11, 12, and 14. [Appendix 6] 4 is further characterized by the particle shape and size depicted in Figures 2 and 3; the crystalline form characterized by the XRPD pattern depicted in Figure 6 is further characterized by the particle shape and size depicted in Figure 5; the crystalline form characterized by the XRPD pattern depicted in Figure 8 is further characterized by the particle shape and size depicted in Figure 7; and the crystalline form characterized by the XRPD pattern depicted in Figure 14 is further characterized by the particle shape and size depicted in Figure 13. [Appendix 7] 2. The crystalline form of claim 1, wherein the salt is a fumarate salt. [Appendix 8] 8. The crystalline form of claim 7, wherein the Fumarate Salt is characterized by the XRPD patterns shown in Figures 29, 30, 32, 33, and 34. [Appendix 9] 29 is further characterized by the particle shape depicted in FIG. 28, and the fumarate salt characterized by the XRPD pattern depicted in FIG. 32 is further characterized by the particle shape depicted in FIG. 31. [Appendix 10] 2. The crystalline form of claim 1, wherein the crystalline form is a free base. [Appendix 11] 11. The crystalline form of claim 10, wherein the free base is characterized by the XRPD pattern shown in Figure 16. [Appendix 12] 12. The crystalline form of claim 11, wherein the crystalline form characterized by the XRPD pattern shown in Figure 16 is further characterized by the particle shape depicted in Figure 15. [Appendix 13] A dosage form comprising a therapeutically effective amount of A2-73 in a crystalline form selected from the group consisting of A2-73 free base and A2-73 salts. [Appendix 14] 14. The dosage form of claim 13, wherein the dosage form comprises about 1 mg to about 50 g, about 1 mg to about 500 mg, or about 1 mg to about 100 mg of A2-73 free base or A2-73 salt. [Appendix 15] 14. The dosage form of claim 13, wherein the dosage form is formulated for extended release of crystalline A2-73. [Appendix 16] 16. The dosage form of claim 15, wherein A2-73 is a free base. [Appendix 17] 17. The dosage form of claim 16, wherein the dosage form comprises from about 1 mg to about 500 mg of A2-73 free base. [Appendix 18] 17. The dosage form of claim 16, wherein the dosage form is a transdermal patch. [Appendix 19] 19. The dosage form of claim 18, wherein the transdermal patch comprises about 40 mg to about 60 mg, about 80 mg to about 120 mg, or about 180 mg to about 220 mg of A2-73 free base. [Appendix 20] 17. The dosage form of claim 16, wherein the dosage form is an enteric-coated oral formulation. [Appendix 21] 21. The dosage form of claim 20, wherein the enteric coated oral formulation comprises about 1 mg to about 50 mg of A2-73 free base. [Appendix 22] 16. The dosage form of claim 15, wherein A2-73 is a salt. [Appendix 23] 23. The dosage form of claim 22, wherein the A2-73 salt is selected from the group consisting of fumarate, sulfate, mesylate, dihydrogen phosphate, edisylate, benzoate, hydrochloride, and oxalate. [Appendix 24] 24. The dosage form of claim 23, wherein the A2-73 salt is a fumarate salt. [Appendix 25] 25. The dosage form of claim 24, wherein the dosage form is a transdermal patch. [Appendix 26] 26. The dosage form of claim 25, wherein the transdermal patch comprises about 1 mg to about 55 mg of A2-73 fumarate. [Appendix 27] 25. The dosage form of claim 24, wherein the dosage form is an enteric coated oral formulation. [Appendix 28] 28. The dosage form of claim 27, wherein the enteric coated oral formulation comprises about 10 mg to about 50 mg, about 20 mg to about 30 mg, or about 15 mg to about 25 mg of A2-73 fumarate. [Appendix 29] A pharmaceutical formulation for the delivery of A2-73, comprising a therapeutically effective amount of a crystalline form of A2-73 selected from A2-73 free base and A2-73 salts. [Appendix 30] 30. The formulation of claim 29, wherein the formulation further comprises one or more pharmaceutically acceptable non-medicinal ingredients selected from chemical enhancers, humectants, pressure-sensitive adhesives, antioxidants, solubilizers, thickeners, plasticizers, adjuvants, carriers, excipients, vehicles, and any combination thereof. [Appendix 31] 31. The formulation of claim 30, wherein the one or more non-medicinal ingredients are selected for oral, transdermal, parenteral, intraperitoneal, intravascular, subcutaneous, by inhalation spray, rectal, or pulmonary administration. [Appendix 32] 30. The formulation of claim 29, wherein the crystalline A2-73 is selected from the free base, the fumarate salt, and the hydrochloride salt. [Appendix 33] 30. The formulation of claim 29, wherein the formulation is an oral formulation comprising about 1% to about 100% by weight of crystalline A2-73. [Appendix 34] 30. The formulation of claim 29, wherein the formulation is for extended delivery of crystalline A2-73. [Appendix 35] 35. The formulation of claim 34, wherein the formulation comprises about 1 mg to about 50 g of crystalline A2-73. [Appendix 36] 35. The formulation of claim 34, wherein the formulation is a subcutaneously injectable formulation containing about 0.5 g to about 3 g of crystalline A2-73. [Appendix 37] 30. The formulation of claim 29, wherein the formulation is a transdermal patch. [Appendix 38] 38. The formulation of claim 37, wherein the patch comprises about 40 mg to about 60 mg, about 80 mg to about 120 mg, or about 180 mg to about 220 mg of A2-73 free base. [Appendix 39] 38. The formulation of claim 37, wherein the patch comprises about 1 mg to about 55 mg of A2-73 fumarate. [Appendix 40] 30. The formulation of claim 29, wherein the formulation is an oral formulation. [Appendix 41] 41. The formulation of claim 40, wherein the oral formulation comprises about 1 mg to about 50 mg of A2-73 free base. [Appendix 42] 41. The formulation of claim 40, wherein the oral formulation comprises about 10 mg to about 50 mg, about 20 mg to about 30 mg, or about 15 mg to about 25 mg of A2-73 fumarate. [Appendix 43] 41. The formulation of claim 40, wherein the oral formulation comprises A2-73 hydrochloride. [Appendix 44] 30. The formulation of claim 29, wherein the formulation is a subcutaneous dosage form containing about 0.1 to about 5 g of crystalline A2-73. [Appendix 45] A method of administering A2-73 to a subject in need thereof, comprising administering the A2-73 to the subject in a dosage form containing a crystalline form of A2-73 selected from A2-73 free base and A2-73 salt. [Appendix 46] 46. ​​The method of claim 45, wherein the crystalline A2-73 is a free base. [Appendix 47] 46. ​​The method of claim 45, wherein the crystalline A2-73 is a fumarate salt. [Appendix 48] 46. ​​The method of claim 45, wherein the crystalline A2-73 is a fumarate salt. [Appendix 49] 46. ​​The method of claim 45, wherein the dosage form is an extended-release transdermal patch and the crystalline A2-73 is administered topically using the transdermal patch. [Appendix 50] 50. The method of claim 49, wherein the transdermal patch is replaced weekly. [Appendix 51] 50. The method of claim 49, wherein the transdermal patch maintains a level of A2-73 in the subject's blood in the range of about 5 ng / ml to about 15 ng / ml, and particularly about 10 ng / ml. [Appendix 52] 46. ​​The method of claim 45, wherein the dosage form is an enteric-coated oral dosage form and the crystalline A2-73 is orally administered using the enteric-coated oral dosage form. [Appendix 53] A method for treating Alzheimer's disease in a subject in need thereof, comprising administering a therapeutically effective amount of a dosage form containing a crystalline form of A2-73 selected from A2-73 free base and A2-73 salt. [Appendix 54] A method for treating progressive dementia in a subject in need thereof, comprising administering a therapeutically effective amount of a dosage form containing a crystalline form of A2-73 selected from A2-73 free base and A2-73 salt. [Appendix 55] A method for treating a neurodegenerative disease in a subject in need thereof, comprising administering to the subject an anti-neurodegenerative effective amount of crystalline A2-73 selected from A2-73 free base and A2-73 salts. [Appendix 56] 53. The method of claim 52, wherein the degenerative disease is selected from Alzheimer's disease, Parkinson's disease, prion diseases, Huntington's disease, motor neuron diseases (MNDs) such as amyotrophic lateral sclerosis, spinocerebellar ataxias (SCAs), and spinal muscular atrophy (SMA). [Appendix 57] 53. The method of claim 52, wherein the anti-neurodegenerative effective amount of A2-73 is about 0.5 mg / day to about 100 mg / day. [Appendix 58] 53. The method of claim 52, wherein the anti-neurodegenerative effective amount of A2-73 is about 1 to about 60 mg / day. [Appendix 59] 53. The method of claim 52, wherein the anti-neurodegenerative effective amount of A2-73 is about 20 to about 50 mg / day. [Appendix 60] 53. The method of claim 52, wherein the anti-neurodegenerative effective amount of A2-73 is about 20 to about 30 mg / day. [Appendix 61] 53. The method of claim 52, wherein the anti-neurodegenerative effective amount of A2-73 is about 15 to about 25 mg / day. [Appendix 62] 53. The method of claim 52, wherein the anti-neurodegenerative effective amount of A2-73 provides a blood level of about 10 ng / ml, about 12 ng / ml, or about A2-73.

Claims

1. A crystalline form of tetrahydro-N,N-dimethyl-2,2-diphenyl-3-furanmethanamine (A2-73), which is a salt or a free base.

2. 10. The crystalline form of claim 1, wherein the crystalline form is a pharmaceutically acceptable salt.

3. 3. The crystalline form of claim 2, wherein the salt is selected from hydrochloride, fumarate, sulfate, dihydrogen phosphate, benzoate, mesylate, edysylate, and oxalate.

4. The crystalline form of claim 3, wherein the salt is a hydrochloride salt.

5. 5. The crystalline form of claim 4, wherein the hydrochloride salt is characterized by the XRPD patterns shown in Figures 4, 6, 8, 9, 10, 11, 12, and 14.

6. 6. The crystalline form of claim 5, wherein the hydrochloride salt characterized by the XRPD pattern shown in FIG. 4 is further characterized by the particle shape and size depicted in FIGS. 2 and 3; the crystalline form characterized by the XRPD pattern shown in FIG. 6 is further characterized by the particle shape and size depicted in FIG. 5; the crystalline form characterized by the XRPD pattern shown in FIG. 8 is further characterized by the particle shape and size depicted in FIG. 7; and the crystalline form characterized by the XRPD pattern shown in FIG. 14 is further characterized by the particle shape and size depicted in FIG.

13.

7. 2. The crystalline form of claim 1, wherein the salt is a fumarate salt.

8. 8. The crystalline form of claim 7, wherein the Fumarate Salt is characterized by the XRPD patterns shown in Figures 29, 30, 32, 33, and 34.

9. 9. The crystalline form of claim 8, wherein the fumarate salt characterized by the XRPD pattern shown in Figure 29 is further characterized by the particle shape illustrated in Figure 28, and the fumarate salt characterized by the XRPD pattern shown in Figure 32 is further characterized by the particle shape illustrated in Figure 31.

10. 2. The crystalline form of claim 1, wherein the crystalline form is a free base.

11. 11. The crystalline form of claim 10, wherein the free base is characterized by the XRPD pattern shown in Figure 16.

12. 12. The crystalline form of claim 11, wherein the crystalline form characterized by the XRPD pattern shown in Figure 16 is further characterized by the particle shape depicted in Figure 15.

13. A dosage form comprising a therapeutically effective amount of A2-73 in a crystalline form selected from the group consisting of A2-73 free base and A2-73 salt.

14. 14. The dosage form of claim 13, wherein the dosage form comprises about 1 mg to about 50 g, about 1 mg to about 500 mg, or about 1 mg to about 100 mg of A2-73 free base or A2-73 salt.

15. 14. The dosage form of claim 13, wherein the dosage form is formulated for extended release of crystalline A2-73. Dosage form.

16. 16. The dosage form of claim 15, wherein A2-73 is a free base.

17. 17. The dosage form of claim 16, wherein the dosage form comprises from about 1 mg to about 500 mg of A2-73 free base.

18. 17. The dosage form of claim 16, wherein the dosage form is a transdermal patch.

19. 19. The dosage form of claim 18, wherein the transdermal patch comprises about 40 mg to about 60 mg, about 80 mg to about 120 mg, or about 180 mg to about 220 mg of A2-73 free base.

20. 17. The dosage form of claim 16, wherein the dosage form is an enteric coated oral formulation.

21. 21. The dosage form of claim 20, wherein the enteric coated oral dosage form comprises from about 1 mg to about 50 mg of A2-73 free base.

22. 16. The dosage form of claim 15, wherein A2-73 is a salt.

23. 23. The dosage form of claim 22, wherein the A2-73 salt is selected from the group consisting of fumarate, sulfate, mesylate, dihydrogen phosphate, edisylate, benzoate, hydrochloride, and oxalate.

24. 24. The dosage form of claim 23, wherein the A2-73 salt is a fumarate salt.

25. 25. The dosage form of claim 24, wherein the dosage form is a transdermal patch.

26. 26. The dosage form of claim 25, wherein the transdermal patch comprises from about 1 mg to about 55 mg of A2-73 fumarate.

27. 25. The dosage form of claim 24, wherein the dosage form is an enteric coated oral formulation.

28. 28. The dosage form of claim 27, wherein the enteric coated oral formulation comprises about 10 mg to about 50 mg, about 20 mg to about 30 mg, or about 15 mg to about 25 mg of A2-73 fumarate.

29. A pharmaceutical formulation for the delivery of A2-73, comprising a therapeutically effective amount of a crystalline form of A2-73 selected from A2-73 free base and A2-73 salts.

30. 30. The formulation of claim 29, wherein the formulation further comprises one or more pharmaceutically acceptable non-medicinal ingredients selected from chemical enhancers, humectants, pressure sensitive adhesives, antioxidants, solubilizers, thickeners, plasticizers, adjuvants, carriers, excipients, vehicles, and any combination thereof.

31. 31. The formulation of claim 30, wherein the one or more non-medicinal ingredients are selected for oral, transdermal, parenteral, intraperitoneal, intravascular, subcutaneous, by inhalation spray, rectal, or pulmonary administration.

32. 30. The formulation of claim 29, wherein the crystalline A2-73 is selected from the free base, the fumarate salt, and the hydrochloride salt. zu

33. 30. The formulation of claim 29, wherein the formulation is an oral formulation comprising about 1% to about 100% by weight of crystalline A2-73.

34. 30. The formulation of claim 29, wherein the formulation is for extended delivery of crystalline A2-73.

35. 35. The formulation of claim 34, wherein the formulation comprises from about 1 mg to about 50 g of crystalline A2-73.

36. 35. The formulation of claim 34, wherein the formulation is a subcutaneous injectable dosage formulation comprising about 0.5 g to about 3 g of crystalline A2-73.

37. 30. The formulation of claim 29, wherein the formulation is a transdermal patch.

38. 38. The formulation of claim 37, wherein the patch comprises about 40 mg to about 60 mg, about 80 mg to about 120 mg, or about 180 mg to about 220 mg of A2-73 free base.

39. 38. The formulation of claim 37, wherein the patch comprises from about 1 mg to about 55 mg of A2-73 fumarate.

40. 30. The formulation of claim 29, wherein the formulation is an oral formulation.

41. 41. The formulation of claim 40, wherein the oral formulation comprises from about 1 mg to about 50 mg of A2-73 free base.

42. 41. The formulation of claim 40, wherein the oral formulation comprises about 10 mg to about 50 mg, about 20 mg to about 30 mg, or about 15 mg to about 25 mg of A2-73 fumarate.

43. 41. The formulation of claim 40, wherein the oral formulation comprises A2-73 hydrochloride.

44. 30. The formulation of claim 29, wherein the formulation is a subcutaneous dosage form comprising about 0.1 to about 5 g of crystalline A2-73.

45. A method of administering A2-73 to a subject in need thereof, comprising administering said A2-73 to said subject in a dosage form comprising a crystalline form of A2-73 selected from A2-73 free base and A2-73 salt.

46. 46. ​​The method of claim 45, wherein the crystalline A2-73 is a free base.

47. 46. ​​The method of claim 45, wherein the crystalline A2-73 is the fumarate salt.

48. 46. ​​The method of claim 45, wherein the crystalline A2-73 is the fumarate salt.

49. 46. ​​The method of claim 45, wherein the dosage form is an extended-release transdermal patch and the crystalline A2-73 is administered topically using the transdermal patch.

50. 50. The method of claim 49, wherein the transdermal patch is replaced weekly.

51. The transdermal patch reduces the level of A2-73 in the subject's blood to between about 5 ng / ml and about 1 50. The method of claim 49, wherein the concentration is maintained in the range of 5 ng / ml, in particular at about 10 ng / ml.

52. 46. ​​The method of claim 45, wherein the dosage form is an enteric coated oral dosage form and the crystalline A2-73 is orally administered using the enteric coated oral dosage form.

53. A method of treating Alzheimer's disease in a subject in need thereof, comprising administering a therapeutically effective amount of a dosage form comprising a crystalline form of A2-73 selected from A2-73 free base and A2-73 salt.

54. A method of treating progressive dementia in a subject in need thereof, comprising administering a therapeutically effective amount of a dosage form comprising a crystalline form of A2-73 selected from A2-73 free base and A2-73 salt.

55. A method of treating a neurodegenerative disease in a subject in need thereof, comprising administering to the subject an anti-neurodegenerative effective amount of crystalline A2-73 selected from A2-73 free base and A2-73 salts.

56. 53. The method of claim 52, wherein the degenerative disease is selected from Alzheimer's disease, Parkinson's disease, prion diseases, Huntington's disease, motor neuron diseases (MNDs) such as amyotrophic lateral sclerosis, spinocerebellar ataxias (SCAs), and spinal muscular atrophy (SMA).

57. 53. The method of claim 52, wherein the anti-neurodegenerative effective amount of A2-73 is from about 0.5 mg / day to about 100 mg / day.

58. 53. The method of claim 52, wherein the anti-neurodegenerative effective amount of A2-73 is from about 1 to about 60 mg / day.

59. 53. The method of claim 52, wherein the anti-neurodegenerative effective amount of A2-73 is from about 20 to about 50 mg / day.

60. 53. The method of claim 52, wherein the anti-neurodegenerative effective amount of A2-73 is about 20 to about 30 mg / day.

61. 53. The method of claim 52, wherein the anti-neurodegenerative effective amount of A2-73 is about 15 to about 25 mg / day.

62. 53. The method of claim 52, wherein the anti-neurodegenerative effective amount of A2-73 provides a blood level of about 10 ng / ml, about 12 ng / ml, about A2-73.

Images