Sublingual compositions for fasudil

Abstract

Disclosed herein are sublingual formulations of fasudil and methods for using the same to treat neurological conditions such as ALS.

Description

SUBLINGUAL COMPOSITIONS FOR FASUDILCROSS REFERENCE

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 553,771 filed February 15, 2024, which is incorporated by reference herein in its entirety.BACKGROUND

[0002] Neurodegenerative diseases, characterized by the progressive loss of structure or function of neurons, affect numerous people around the world. For example, amyotrophic lateral sclerosis, also known as ALS, is a disease that affects nerve cells in the brain and spinal cord, eventually causing loss of muscle control. Early death is common as the loss of muscle control eventually hinders vital functions, such as breathing.INCORPORATION BY REFERENCE

[0003] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.SUMMARY OF THE INVENTION

[0004] In some embodiments disclosed herein are compositions comprising fasudil or a pharmaceutically acceptable salt thereof, wherein the composition is a sublingual formulation. In some embodiments disclosed herein are methods for treating amyotrophic lateral sclerosis (ALS), the methods comprising sublingually administering to a subject in need thereof a therapeutically-effective amount of fasudil or a pharmaceutically acceptable salt thereof.

[0005] In some embodiments disclosed herein are pharmaceutical compositions comprising fasudil or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition is a sublingual dosage form.

[0006] In some embodiments disclosed herein are pharmaceutical compositions comprising fasudil or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition is a solid dosage form, wherein if the solid dosage form is subjected to a shake test in which the solid dosage form is placed in a glass beaker containing 2 mL of water and the beaker ismanually shaken to disintegrate the solid dosage form in the water, then the time needed for disintegration of the solid dosage form into fine particles is less than sixty seconds.

[0007] In some embodiments disclosed herein are pharmaceutical compositions comprising fasudil or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition is a solid dosage form, wherein if a sample of about 8 mg to about 13 mg of the solid dosage form is subjected to differential thermal analysis by placing the sample in a platinum pan of 100 pL capacity, and the sample is heated from 25 °C to 600 °C at a rate of 10 °C / min under air flow, then a resulting thermograph shows an endothermic peak at 145-170 °C and does not show an endothermic peak at 210-235 °C.

[0008] In some embodiments disclosed herein are methods for treating a condition, the method comprising sublingually administering to a subject in need thereof a therapeutically - effective amount of fasudil or a pharmaceutically acceptable salt thereof.FIGURE DESCRIPTIONS

[0009] Figure 1 shows an example of chemical synthesis of fasudil hydrochloride.

[0010] Figure 2A shows aJH NMR spectrum of fasudil. Figure 2B shows an IR spectrum of fasudil.

[0011] Figure 3 shows a chromatogram and chromatographic parameters of a fasudil peak at 216 nm.

[0012] Figures 4A-4B show the major peak in the 2D (Figure 4A) and 3D (Figure 4B) UV spectra of fasudil. The spectra were recorded from 200 nm to 500 nm.

[0013] Figures 5A-5B show chromatograms of fasudil standard solution (Figure 5A) and fasudil standard solution in the presence of excipients (Figure 5B).

[0014] Figures 6A-6B show the linearity of fasudil standard solutions (Figure 6A) and reconstituted standard solutions (Figure 6B).

[0015] Figure 7 illustrates an example schematic diagram of the in vitro dissolution method.

[0016] Figure 8 shows an image of the sublingual FSD tablets.

[0017] Figure 9 shows a graph of weight loss during differential thermal analysis measurements of fasudil hydrochloride, excipients, and formulated fasudil.

[0018] Figure 10 shows thermograms recorded during differential thermal analysis measurements of fasudil hydrochloride, excipients, and formulated fasudil.

[0019] Figure 11 shows weight variation of fasudil sublingual tablets.

[0020] Figure 12 shows the content uniformity of fasudil tablets.

[0021] Figure 13 shows in vitro dissolution of fasudil tablets.

[0022] Figure 14 shows an example schematic of a cell culture insert.

[0023] Figure 15 shows an example schematic of a cell preparation method.

[0024] Figures 16A-16C show the sample preparation of a fasudil solution (Figure 16A), a fasudil tablet (Figure 16B), and a placebo tablet (Figure 16C).

[0025] Figure 17A shows a plate plan for a bioavailability assay. Figure 17B shows another plate plan for a bioavailability assay. Figure 17C shows an additional plate plan for a bioavailability assay.

[0026] Figure 18A shows resistance (R) and TEER values calculated for each condition of the bioavailability assay for unfiltered samples. Figure 18B shows resistance (R) and TEER values calculated for each condition of the bioavailability assay for filtered samples.

[0027] Figures 19A-19B show the kinetics of the transfer of fasudil through a SCC-4 cell monolayer for unfiltered (Figure 19A) and filtered (Figure 19B) samples.

[0028] Figure 20A shows fasudil transepithelial transfer through SCC-4 cell monolayer versus time for unfiltered samples with an initial apical concentration of fasudil = 200 mg / L. Figure 20B shows fasudil transepithelial transfer through SCC-4 cell monolayer versus time for unfiltered samples with an initial apical concentration of fasudil = 1,000 mg / L. Figure 20C shows fasudil transepithelial transfer through SCC-4 cell monolayer versus time for filtered samples with an initial apical concentration of fasudil = 1,000 mg / L.DETAILED DESCRIPTION OF THE INVENTION

[0029] Amyotrophic lateral sclerosis (ALS) is a fatal disorder characterized by subtle onset of focal weakness, typically in the limbs but sometimes in bulbar muscles. ALS progresses to paralysis of almost all skeletal muscles. Significant clinico-pathological and genetic overlap exists between ALS and frontotemporal lobar dementia (FTLD). In ALS, death from respiratory paralysis is common within five years. The cellular pathology is focal at onset and spreads in a pattern and suggests successive involvement of contiguous neuronal populations. Death of motor neurons occurs in conjunction with deposition of aggregated proteins in motoneurons and oligodendrocytes, and neuroinflammation. While most cases of ALS are sporadic (SALS), about 10% are inherited, usually dominantly (familial ALS, FALS).

[0030] TDP-43 (Transactivating response element DNA binding protein 43 kDa) accumulates in cytoplasm of motor neurons in most cases of ALS. TDP-43 is a nuclear RNA-binding protein involved in several aspects of RNA processing that actively shuttles between the nucleus and cytoplasm. In ALS and frontotemporal dementia, TDP-43 is excluded from the nucleus. However, such cytoplasmic mislocalization is common inneuronal injury or stress. TDP-43 -positive inclusions represent secondary pathology in some neurodegenerative disorders. Possible mechanisms of death of motor neurons in ALS include a) glutamate-mediated excitotoxicity; b) decrease in neurotrophic factors (BDNF, GDNF) and associated signaling; c) mitochondrial alterations and oxidative damages; and d) abnormalities in cytoskeletal proteins resulting in neuronal atrophy and death.

[0031] Formulations of fasudil compositions for the treatment of neurodegenerative diseases (e.g., ALS) can be difficult to administer due to symptoms such as muscle twitches and difficulty chewing or swallowing. In contrast, a sublingual formulation of fasudil allows for treatment of neurodegenerative diseases (e.g., ALS) by allowing for administration.Disclosed herein are compositions comprising fasudil or a pharmaceutically acceptable salt thereof, wherein the compositions are a sublingual formulation, and methods of use thereof.Compounds of the Disclosure

[0032] Described herein are compositions of fasudil or a pharmaceutically-acceptable salt thereof. Fasudil is a nonspecific RhoA / ROCK inhibitor that can have an inhibitory effect on protein kinases. Fasudil is also a calcium channel antagonist that primarily acts through the inhibition of the Rho-kinase signaling pathway. This action allows for vasodilation through the activation of myosin phosphatase. Fasudil is an isoquinoline substituted with a (l,4-diazepan-l-yl)sulfonyl group at position 5. Fasudil has a molecular formula of C14H17N3O2S and a structure of:FasudilPharmaceutically-Acceptable Salts

[0033] The invention provides the use of pharmaceutically-acceptable salts of any compound described herein. Pharmaceutically-acceptable salts include, for example, acid-addition salts and base-addition salts. The acid that is added to the compound to form an acid-addition salt can be an organic acid or an inorganic acid. A base that is added to the compound to form a base-addition salt can be an organic base or an inorganic base. In some embodiments, a pharmaceutically-acceptable salt is a metal salt. In some embodiments, apharmaceutically-acceptable salt is an ammonium salt.

[0034] Metal salts can arise from the addition of an inorganic base to a compound of the invention. The inorganic base consists of a metal cation paired with a basic counterion, such as, for example, hydroxide, carbonate, bicarbonate, or phosphate. The metal can be an alkali metal, alkaline earth metal, transition metal, or main group metal. In some embodiments, the metal is lithium, sodium, potassium, cesium, cerium, magnesium, manganese, iron, calcium, strontium, cobalt, titanium, aluminum, copper, cadmium, or zinc.

[0035] In some embodiments, a metal salt is a lithium salt, a sodium salt, a potassium salt, a cesium salt, a cerium salt, a magnesium salt, a manganese salt, an iron salt, a calcium salt, a strontium salt, a cobalt salt, a titanium salt, an aluminum salt, a copper salt, a cadmium salt, or a zinc salt.

[0036] Ammonium salts can arise from the addition of ammonia or an organic amine to a compound of the invention. In some embodiments, the organic amine is triethyl amine, diisopropyl amine, ethanol amine, diethanol amine, triethanol amine, morpholine, N-methylmorpholine, piperidine, N-methylpiperidine, N-ethylpiperidine, dibenzylamine, piperazine, pyridine, pyrazole, piprazole, imidazole, or pyrazine.

[0037] In some embodiments, an ammonium salt is a tri ethyl amine salt, a diisopropyl amine salt, an ethanol amine salt, a diethanol amine salt, a triethanol amine salt, a morpholine salt, an N-methylmorpholine salt, a piperidine salt, an N-methylpiperidine salt, an N-ethylpiperidine salt, a dibenzylamine salt, a piperazine salt, a pyridine salt, a pyrazole salt, a piprazole salt, an imidazole salt, or a pyrazine salt.

[0038] Acid addition salts can arise from the addition of an acid to a compound of the invention. In some embodiments, the acid is organic. In some embodiments, the acid is inorganic. In some embodiments, the acid is hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, nitrous acid, sulfuric acid, sulfurous acid, a phosphoric acid, isonicotinic acid, lactic acid, salicylic acid, tartaric acid, ascorbic acid, gentisinic acid, gluconic acid, glucaronic acid, saccharic acid, formic acid, benzoic acid, glutamic acid, pantothenic acid, acetic acid, propionic acid, butyric acid, fumaric acid, succinic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, oxalic acid, or maleic acid.

[0039] In some embodiments, the salt is a hydrochloride salt, a hydrobromide salt, a hydroiodide salt, a nitrate salt, a nitrite salt, a sulfate salt, a sulfite salt, a phosphate salt, isonicotinate salt, a lactate salt, a salicylate salt, a tartrate salt, an ascorbate salt, a gentisinate salt, a gluconate salt, a glucaronate salt, a saccharate salt, a formate salt, a benzoate salt, aglutamate salt, a pantothenate salt, an acetate salt, a propionate salt, a butyrate salt, a fumarate salt, a succinate salt, a methanesulfonate salt, an ethanesulfonate salt, a benzenesulfonate salt, a p-toluenesulfonate salt, a citrate salt, an oxalate salt, or a maleate salt.

[0040] In some embodiments, the salt is a hydrochloride salt, forming fasudil hydrochloride. Fasudil hydrochloride has a chemical formula of C14H17N3O2S • HC1 and a structure of:Formulations

[0041] A pharmaceutical composition of the disclosure can provide a therapeutically-effective amount of fasudil. In some embodiments, fasudil or pharmaceutically-acceptable salt thereof is present in a formulation in an amount of from about 0.1 mg / mL to about 100 mg / mL, from about 0.1 mg / mL to about 1 mg / mL, from about 0.1 mg / mL to about 5 mg / mL, from about 5 mg / mL to about 10 mg / mL, from about 10 mg / mL to about 15 mg / mL, from about 15 mg / mL to about 20 mg / mL, from about 20 mg / mL to about 25 mg / mL, from about 25 mg / mL to about 30 mg / mL, from about 30 mg / mL to about 35 mg / mL, from about 35 mg / mL to about 40 mg / mL, from about 40 mg / mL to about 45 mg / mL, about 45 mg / mL to about 50 mg / mL, from about 50 mg / mL to about 55 mg / mL, from about 55 mg / mL to about 60 mg / mL, from about 60 mg / mL to about 65 mg / mL, from about 65 mg / mL to about 70 mg / mL, from about 70 mg / mL to about 75 mg / mL, about 75 mg / mL to about 80 mg / mL, from about 80 mg / mL to about 85 mg / mL, from about 85 mg / mL to about 90 mg / mL, from about 90 mg / mL to about 95 mg / mL, or from about 95 mg / mL to about 100 mg / mL.

[0042] In some embodiments, fasudil or a pharmaceutically-acceptable salt thereof is present in a formulation in an amount of about 1 mg / mL, about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, about 5 mg / mL, about 6 mg / mL, about 7 mg / mL, about 8 mg / mL, about 9 mg / mL, about 10 mg / mL, about 11 mg / mL, about 12 mg / mL, about 13 mg / mL, about 14 mg / mL, about 15 mg / mL, about 16 mg / mL, about 17 mg / mL, about 18 mg / mL, about 19 mg / mL, about 20 mg / mL, about 21 mg / mL, about 22 mg / mL, about 23 mg / mL, about 24 mg / mL, about 25 mg / mL, about 26 mg / mL, about 27 mg / mL, about 28 mg / mL, about 29 mg / mL,about 30 mg / mL, about 31 mg / mL, about 32 mg / mL, about 33 mg / mL, about 34 mg / mL, about 35 mg / mL, about 36 mg / mL, about 37 mg / mL, about 38 mg / mL, about 39 mg / mL, about 40 mg / mL, about 41 mg / mL, about 42 mg / mL, about 43 mg / mL, about 44 mg / mL, about 45 mg / mL, about 46 mg / mL, about 47 mg / mL, about 48 mg / mL, about 49 mg / mL, about 50 mg / mL, about 51 mg / mL, about 52 mg / mL, about 53 mg / mL, about 54 mg / mL, about 55 mg / mL, about 56 mg / mL, about 57 mg / mL, about 58 mg / mL, about 59 mg / mL, about 60 mg / mL, about 61 mg / mL, about 62 mg / mL, about 63 mg / mL, about 64 mg / mL, about 65 mg / mL, about 66 mg / mL, about 67 mg / mL, about 68 mg / mL, about 69 mg / mL, about 70 mg / mL, about 71 mg / mL, about 72 mg / mL, about 73 mg / mL, about 74 mg / mL, about 75 mg / mL, about 76 mg / mL, about 77 mg / mL, about 78 mg / mL, about 79 mg / mL, about 80 mg / mL, about 81 mg / mL, about 82 mg / mL, about 83 mg / mL, about 84 mg / mL, about 85 mg / mL, about 86 mg / mL, about 87 mg / mL, about 88 mg / mL, about 89 mg / mL, about 90 mg / mL, about 91 mg / mL, about 92 mg / mL, about 93 mg / mL, about 94 mg / mL, about 95 mg / mL, about 96 mg / mL, about 97 mg / mL, about 98 mg / mL, about 99 mg / mL, or about 100 mg / mL.

[0043] In some embodiments, fasudil or a pharmaceutically-acceptable salt thereof is present in a formulation in an amount of from about 0.1 mg to about 100 mg, from about 10 mg to about 100 mg, from about 10 mg to about 80 mg, from about 20 mg to about 100 mg, from about 20 mg to about 60 mg, from about 20 mg to about 40 mg, from about 1 mg to about 20 mg, from about 40 mg to about 50 mg, from about 0.1 mg to about 1 mg, from about 0.5 mg to about 1 mg, from about 0.5 mg to about 10 mg, from about 0.1 mg to about 5 mg, from about 5 mg to about 10 mg, from about 10 mg to about 15 mg, from about 15 mg to about 20 mg, from about 20 mg to about 25 mg, from about 25 mg to about 30 mg, from about 30 mg to about 35 mg, from about 35 mg to about 40 mg, from about 40 mg to about 45 mg, about 45 mg to about 50 mg, from about 50 mg to about 55 mg, from about 55 mg to about 60 mg, from about 60 mg to about 65 mg, from about 65 mg to about 70 mg, from about 70 mg to about 75 mg, about 75 mg to about 80 mg, from about 80 mg to about 85 mg, from about 85 mg to about 90 mg, from about 90 mg to about 95 mg, or from about 95 mg to about 100 mg.

[0044] In some embodiments, fasudil or a pharmaceutically-acceptable salt thereof is present in a formulation in an amount of about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, about 10 mg, about 11 mg, about 12 mg, about 13 mg, about 14 mg, about 15 mg, about 16 mg, about 17 mg, about 18 mg, about 19 mg, about 20 mg, about 21 mg, about 22 mg, about 23 mg, about 24 mg, about 25 mg, about 26 mg, about 27 mg, about 28 mg, about 29 mg, about 30 mg, about 31 mg, about 32mg, about 33 mg, about 34 mg, about 35 mg, about 36 mg, about 37 mg, about 38 mg, about 39 mg, about 40 mg, about 41 mg, about 42 mg, about 43 mg, about 44 mg, about 45 mg, about 46 mg, about 47 mg, about 48 mg, about 49 mg, about 50 mg, about 51 mg, about 52 mg, about 53 mg, about 54 mg, about 55 mg, about 56 mg, about 57 mg, about 58 mg, about 59 mg, about 60 mg, about 61 mg, about 62 mg, about 63 mg, about 64 mg, about 65 mg, about 66 mg, about 67 mg, about 68 mg, about 69 mg, about 70 mg, about 71 mg, about 72 mg, about 73 mg, about 74 mg, about 75 mg, about 76 mg, about 77 mg, about 78 mg, about 79 mg, about 80 mg, about 81 mg, about 82 mg, about 83 mg, about 84 mg, about 85 mg, about 86 mg, about 87 mg, about 88 mg, about 89 mg, about 90 mg, about 91 mg, about 92 mg, about 93 mg, about 94 mg, about 95 mg, about 96 mg, about 97 mg, about 98 mg, about 99 mg, or about 100 mg.

[0045] In some embodiments, fasudil or a pharmaceutically-acceptable salt thereof is present in a formulation in a concentration of at least about 10 nM, at least about 20 nM, at least about 30 nM, at least about 40 nM, at least about 50 nM, at least about 60 nM, at least about 70 nM, at least about 80 nM, at least about 90 nM, at least about 100 nM, at least about 150 nM, at least about 200 nM, at least about 250 nM, at least about 300 nM, at least about 350 nM, at least about 400 nM, at least about 450 nM, at least about 500 nM, at least about 550 nM, at least about 600 nM, at least about 650 nM, at least about 700 nM, at least about 750 nM, at least about 800 nM, at least about 950 nM, at least about 1 pM, at least about 2 pM, at least about 3 pM, at least about 4 pM, at least about 5 pM, at least about 6 pM, at least about 7 pM, at least about 8 pM, at least about 9 pM, at least about 10 pM, at least about 11 pM, at least about 12 pM, at least about 13 pM, at least about 14 pM, at least about 15 pM, at least about 16 pM, at least about 17 pM, at least about 18 pM, at least about 19 pM, or at least about 20 pM.

[0046] In some embodiments, fasudil or a pharmaceutically-acceptable salt thereof is present in a formulation in a concentration of at most about 20 pM, at most about 19 pM, at most about 18 pM, at most about 17 pM, at most about 16 pM, at most about 15 pM, at most about 14 pM, at most about 13 pM, at most about 12 pM, at most about 11 pM, at most about 10 pM, at most about 9 pM, at most about 8 pM, at most about 7 pM, at most about 6 pM, at most about 5 pM, at most about 4 pM, at most about 3 pM, at most about 2 pM, at most about 1 pM, at most about 950 nM, at most about 900 nM, at most about 850 nM, at most about 800 nM, at most about 750 nM, at most about 700 nM, at most about 650 nM, at most about 600 nM, at most about 550 nM, at most about 500 nM, at most about 450 nM, at most about 400 nM, at most about 350 nM, at most about 300 nM, at most about 250 nM, at mostabout 200 nM, at most about 150 nM, at most about 100 nM, at most about 90 nM, at most about 80 nM, at most about 70 nM, at most about 60 nM, at most about 50 nM, at most about 40 nM, at most about 30 nM, at most about 20 nM, or at most about 10 nM.

[0047] In some embodiments, fasudil or a pharmaceutically-acceptable salt thereof is present in a formulation in a concentration between 20 nM to 500 nM. In some embodiments, or pharmaceutically-acceptable salt thereof is present in a formulation in a concentration between 50 nM to 300 nM. In some embodiments, fasudil or a pharmaceutically-acceptable salt thereof is present in a formulation in a concentration between 70 nM to 200 nM.

[0048] A formulation that is disclosed herein can be made more soluble by the addition of an additive or agent. A non-limiting example of a solubilizing agent includes an organic solvent. Non-limiting examples of organic solvents include alcohols, for example, C1-C4 linear alkyl, C3-C4 branched alkyl, ethanol, ethylene glycol, glycerin, 2-hydroxypropanol, propylene glycol, maltitol, sorbitol, xylitol, substituted or unsubstituted aryl, and benzyl alcohol.

[0049] A formulation as disclosed herein can comprise a therapeutic compound (e.g., fasudil or a pharmaceutically-acceptable salt thereof) and an additive in a ratio of 1: 1. For example, a formulation containing about 100 mg of a therapeutic compound can comprise about 100 mg of an additive. In another embodiment, the ratio can be based on the number, or moles, of a therapeutic compound compared to the number, or moles, of an additive. Additional nonlimiting ratios of a therapeutic compound (e.g., fasudil or a pharmaceutically-acceptable salt thereof) and an additive can be about 20:about 1; about 19.9:about 1; about 19.8:about 1; about 19.7:about 1; about 19.6:about 1; about 19.5:about 1; about 19.4:about 1; about 19.3:about 1; about 19.2:about 1; about 19.1:about 1; about 19:about 1; about 18.9:about 1; about 18.8:about 1; about 18.7:about 1; about 18.6:about 1; about 18.5:about 1; about 18.4:about 1; about 18.3:about 1; about 18.2:about 1; about 18.1:about 1; about 18:about 1; about 17.9:about 1; about 17.8:about 1; about 17.7:about 1; about 17.6:about 1; about 17.5:about 1; about 17.4:about 1; about 17.3:about 1; about 17.2:about 1; about 17.1 :about 1; about 17:about 1; about 16.9:about 1; about 16.8:about 1; about 16.7:about 1; about 16.6:about 1; about 16.5:about 1; about 16.4:about 1; about 16.3:about 1; about 16.2:about 1; about 16.1 : about 1; about 16: about 1; about 15.9: about 1; about 15.8: about 1; about 15.7:about 1; about 15.6:about 1; about 15.5:about 1; about 15.4:about 1; about 15.3:about 1; about 15.2:about 1; about 15.1 :about 1; about 15:about 1; about 14.9:about 1; about 14.8:about 1; about 14.7:about 1; about 14.6:about 1; about 14.5:about 1; about 14.4:about 1; about 14.3:about 1; about 14.2: about 1; about 14.1:about 1; about 14:about 1; about 13.9:about 1; about 13.8:about 1; about 13.7:about 1; about 13.6:about 1; about 13.5:about 1;about 13.4:about 1; about 13.3:about 1; about 13.2:about 1; about 13.1 :about 1; about 13:about 1; about 12.9:about 1; about 12.8:about 1; about 12.7:about 1; about 12.6:about 1; about 12.5:about 1; about 12.4:about 1; about 12.3:about 1; about 12.2:about 1; about 12.1 :about 1; about 12:about 1; about 11.9:about 1; about 11.8:about 1; about 11.7:about 1; about 11.6:about 1; about 11.5:about 1; about 11.4:about 1; about 11.3:about 1; about 11.2:about 1; about l l.l :about 1; about l l :about 1; about 10.9:about 1; about 10.8:about 1; about 10.7:about 1; about 10.6:about 1; about 10.5:about 1; about 10.4:about 1; about 10.3:about 1; about 10.2:about 1; about 10.1:about 1; about 10:about 1; about 9.9:about 1; about 9.8:about 1; about 9.7:about 1; about 9.6:about 1; about 9.5:about 1; about 9.4:about 1; about 9.3:about 1; about 9.2:about 1; about 9.1 :about 1; about 9:about 1; about 8.9:about 1; about 8.8:about 1; about 8.7:about 1; about 8.6:about 1; about 8.5:about 1; about 8.4:about 1; about 8.3:about 1; about 8.2:about 1; about 8.1 :about 1; about 8:about 1; about 7.9:about 1; about 7.8:about 1; about 7.7:about 1; about 7.6:about 1; about 7.5:about 1; about 7.4:about 1; about 7.3:about 1; about 7.2:about 1; about 7.1 :about 1; about 7:about 1; about 6.9:about 1; about 6.8:about 1; about 6.7:about 1; about 6.6:about 1; about 6.5:about 1; about 6.4:about 1; about 6.3:about 1; about 6.2:about 1; about 6.1 :about 1; about 6:about 1; about 5.9:about 1; about 5.8:about 1; about 5.7:about 1; about 5.6:about 1; about 5.5:about 1; about 5.4:about 1; about 5.3:about 1; about 5.2:about 1; about 5.1 :about 1; about 5:about 1; about 4.9:about 1; about 4.8:about 1; about 4.7:about 1; about 4.6:about 1; about 4.5:about 1; about 4.4:about 1; about 4.3:about 1; about 4.2:about 1; about 4.1 :about 1; about 4:about 1; about 3.9:about 1; about 3.8:about 1; about 3.7:about 1; about 3.6:about 1; about 3.5:about 1; about 3.4:about 1; about 3.3:about 1; about 3.2:about 1; about 3.1 :about 1; about 3:about 1; about 2.9:about 1; about 2.8:about 1; about 2.7:about 1; about 2.6:about 1; about 2.5:about 1; about 2.4:about 1; about 2.3:about 1; about 2.2:about 1; about 2.1 :about 1; about 2:about 1; about 1.9:about 1; about 1.8:about 1; about 1.7:about 1; about 1.6:about 1; about 1.5:about 1; about 1.4:about 1; about 1.3:about 1; about 1.2:about 1; about l. l :about 1; or about l :about 1.

[0050] An additive or agent can increase the solubility of the formulation by about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75% about 80%, about 85%, about 90%, about 95%, about 100%, about 110%, about 120%, about 130%, about 140%, about 150%, about 160%, about 170%, about 180%, about 190%, about 200%, about 225%, about 250%, about 275%, about 300%, about 325%, about 350%, about 375%, about 400%, about 450%, or about 500%.

[0051] A formulation disclosed herein can be stable for about 1 day, about 2 days, about 3days, about 4 days, about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 2 weeks, about 4 weeks, about 6 weeks, about 8 weeks, about 10 weeks, about 12 weeks, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, or about one year. A formulation disclosed herein can be stable, for example, at about 0 °C, about 5 °C, about 10 °C, about 15 °C, about 20 °C, about 25 °C, about 30 °C, about 35 °C, about 40 °C, about 45 °C, about 50 °C, about 60 °C, about 70 °C, or about 80 °C.

[0052] A pharmaceutical composition of the invention can be a combination of any pharmaceutical compounds described herein with other chemical components, such as carriers, stabilizers, diluents, dispersing agents, suspending agents, thickening agents, or excipients. Pharmaceutical compositions can be formulated using one or more physiologically-acceptable carriers comprising excipients and auxiliaries, which facilitate processing of the active compounds into preparations that can be used pharmaceutically. Formulations can be modified depending upon the route of administration chosen.Pharmaceutical compositions comprising a compound described herein can be manufactured, for example, by mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping, or compression processes.

[0053] The pharmaceutical compositions can include at least one pharmaceutically- acceptable carrier, diluent, or excipient and compounds described herein as free-base or pharmaceutically-acceptable salt form. The methods and pharmaceutical compositions described herein include the use of crystalline forms (also known as polymorphs), and active metabolites of these compounds having the same type of activity.

[0054] A pharmaceutical composition can comprise one or more carriers. Non-limiting examples of pharmaceutically-acceptable carriers include saline solution, Ringer's solution, and dextrose solution. The pH of the solution can be from about 5 to about 8. The pH of the solution can be from about 7 to about 7.5. Further carriers include sustained release preparations such as semipermeable matrices of solid hydrophobic polymers containing the fasudil or a pharmaceutically-acceptable salt thereof, where the matrices are in the form of shaped articles, such as films, liposomes, microparticles, and microcapsules.

[0055] A pharmaceutical composition can comprise one or more fillers. Non-limiting examples of fillers can include lactose, sucrose, magnesium stearate, glucose, cellulose, and calcium carbonate. In some embodiments, a filler can be cellulose (e.g., microcrystaHine cellulose).

[0056] In some embodiments, a filler (e.g., a cellulose or microcrystalline cellulose) ispresent in a composition in an amount of present in a formulation in an amount of from about 0.1 mg to about 100 mg, from about 10 mg to about 100 mg, from about 10 mg to about 80 mg, from about 20 mg to about 100 mg, from about 20 mg to about 60 mg, from about 20 mg to about 40 mg, from about 1 mg to about 20 mg, from about 40 mg to about 50 mg, from about 0. 1 mg to about 1 mg, from about 0.5 mg to about 1 mg, from about 0.5 mg to about 10 mg, from about 0.1 mg to about 5 mg, from about 5 mg to about 10 mg, from about 10 mg to about 15 mg, from about 15 mg to about 20 mg, from about 20 mg to about 25 mg, from about 25 mg to about 30 mg, from about 30 mg to about 35 mg, from about 35 mg to about 40 mg, from about 40 mg to about 45 mg, about 45 mg to about 50 mg, from about 50 mg to about 55 mg, from about 55 mg to about 60 mg, from about 60 mg to about 65 mg, from about 65 mg to about 70 mg, from about 70 mg to about 75 mg, about 75 mg to about 80 mg, from about 80 mg to about 85 mg, from about 85 mg to about 90 mg, from about 90 mg to about 95 mg, or from about 95 mg to about 100 mg.

[0057] In some embodiments, a filler (e.g., cellulose or microcrystalline cellulose) is present in a formulation in an amount of about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, about 10 mg, about 11 mg, about 12 mg, about 13 mg, about 14 mg, about 15 mg, about 16 mg, about 17 mg, about 18 mg, about 19 mg, about 20 mg, about 21 mg, about 22 mg, about 23 mg, about 24 mg, about 25 mg, about 26 mg, about 27 mg, about 28 mg, about 29 mg, about 30 mg, about 31 mg, about 32 mg, about 33 mg, about 34 mg, about 35 mg, about 36 mg, about 37 mg, about 38 mg, about 39 mg, about 40 mg, about 41 mg, about 42 mg, about 43 mg, about 44 mg, about 45 mg, about 46 mg, about 47 mg, about 48 mg, about 49 mg, about 50 mg, about 51 mg, about 52 mg, about 53 mg, about 54 mg, about 55 mg, about 56 mg, about 57 mg, about 58 mg, about 59 mg, about 60 mg, about 61 mg, about 62 mg, about 63 mg, about 64 mg, about 65 mg, about 66 mg, about 67 mg, about 68 mg, about 69 mg, about 70 mg, about 71 mg, about 72 mg, about 73 mg, about 74 mg, about 75 mg, about 76 mg, about 77 mg, about 78 mg, about 79 mg, about 80 mg, about 81 mg, about 82 mg, about 83 mg, about 84 mg, about 85 mg, about 86 mg, about 87 mg, about 88 mg, about 89 mg, about 90 mg, about 91 mg, about 92 mg, about 93 mg, about 94 mg, about 95 mg, about 96 mg, about 97 mg, about 98 mg, about 99 mg, or about 100 mg.

[0058] A pharmaceutical composition can comprise one or more binders. Non-limiting examples of binders can include starch, sodium alginate, gelatin, polyvinyl pyrrolidone (PVP), methylcellulose, hydroxy propyl methyl cellulose (PHMC), polymethacrylate, sodium carboxy methyl cellulose, polyethylene glycol (PEG), methylcellulose, lactose, sucrose,sorbitol, xylitol, or mannitol. In some embodiments, a binder can be mannitol.

[0059] In some embodiments, a binder (e.g., mannitol) is present in a formulation in an amount of from about 0. 1 mg to about 100 mg, from about 10 mg to about 100 mg, from about 10 mg to about 80 mg, from about 20 mg to about 100 mg, from about 20 mg to about60 mg, from about 20 mg to about 40 mg, from about 1 mg to about 20 mg, from about 40 mg to about 50 mg, from about 0. 1 mg to about 1 mg, from about 0.5 mg to about 1 mg, from about 0.5 mg to about 10 mg, from about 0. 1 mg to about 5 mg, from about 5 mg to about 10 mg, from about 10 mg to about 15 mg, from about 15 mg to about 20 mg, from about 20 mg to about 25 mg, from about 25 mg to about 30 mg, from about 30 mg to about 35 mg, from about 35 mg to about 40 mg, from about 40 mg to about 45 mg, about 45 mg to about 50 mg, from about 50 mg to about 55 mg, from about 55 mg to about 60 mg, from about 60 mg to about 65 mg, from about 65 mg to about 70 mg, from about 70 mg to about 75 mg, about 75 mg to about 80 mg, from about 80 mg to about 85 mg, from about 85 mg to about 90 mg, from about 90 mg to about 95 mg, or from about 95 mg to about 100 mg.

[0060] In some embodiments, a binder (e.g., mannitol) is present in a formulation in an amount of about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, about 10 mg, about 11 mg, about 12 mg, about 13 mg, about 14 mg, about 15 mg, about 16 mg, about 17 mg, about 18 mg, about 19 mg, about 20 mg, about 21 mg, about 22 mg, about 23 mg, about 24 mg, about 25 mg, about 26 mg, about 27 mg, about 28 mg, about 29 mg, about 30 mg, about 31 mg, about 32 mg, about 33 mg, about 34 mg, about 35 mg, about 36 mg, about 37 mg, about 38 mg, about 39 mg, about 40 mg, about 41 mg, about 42 mg, about 43 mg, about 44 mg, about 45 mg, about 46 mg, about 47 mg, about 48 mg, about 49 mg, about 50 mg, about 51 mg, about 52 mg, about 53 mg, about 54 mg, about 55 mg, about 56 mg, about 57 mg, about 58 mg, about 59 mg, about 60 mg, about61 mg, about 62 mg, about 63 mg, about 64 mg, about 65 mg, about 66 mg, about 67 mg, about 68 mg, about 69 mg, about 70 mg, about 71 mg, about 72 mg, about 73 mg, about 74 mg, about 75 mg, about 76 mg, about 77 mg, about 78 mg, about 79 mg, about 80 mg, about 81 mg, about 82 mg, about 83 mg, about 84 mg, about 85 mg, about 86 mg, about 87 mg, about 88 mg, about 89 mg, about 90 mg, about 91 mg, about 92 mg, about 93 mg, about 94 mg, about 95 mg, about 96 mg, about 97 mg, about 98 mg, about 99 mg, or about 100 mg.

[0061] A pharmaceutical composition can comprise one or more superdisintegrants. Nonlimiting examples of superdisintegrants can include croscarmellose, crospovidone, sodium starch glycolate, or magnesium aluminum silicate. In some embodiments, a superdisintegrant can be sodium starch glycolate.

[0062] In some embodiments, a superdisintegrant (e.g., sodium starch glycolate) is present in a formulation in an amount of from about 0.1 mg to about 100 mg, from about 10 mg to about 100 mg, from about 10 mg to about 80 mg, from about 20 mg to about 100 mg, from about 20 mg to about 60 mg, from about 20 mg to about 40 mg, from about 1 mg to about 20 mg, from about 40 mg to about 50 mg, from about 0. 1 mg to about 1 mg, from about 0.5 mg to about 1 mg, from about 0.5 mg to about 10 mg, from about 0. 1 mg to about 5 mg, from about 5 mg to about 10 mg, from about 10 mg to about 15 mg, from about 15 mg to about 20 mg, from about 20 mg to about 25 mg, from about 25 mg to about 30 mg, from about 30 mg to about 35 mg, from about 35 mg to about 40 mg, from about 40 mg to about 45 mg, about45 mg to about 50 mg, from about 50 mg to about 55 mg, from about 55 mg to about 60 mg, from about 60 mg to about 65 mg, from about 65 mg to about 70 mg, from about 70 mg to about 75 mg, about 75 mg to about 80 mg, from about 80 mg to about 85 mg, from about 85 mg to about 90 mg, from about 90 mg to about 95 mg, or from about 95 mg to about 100 mg.

[0063] In some embodiments, a superdisintegrant (e.g., sodium starch glycolate) is present in a formulation in an amount of about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, about 10 mg, about 11 mg, about 12 mg, about 13 mg, about 14 mg, about 15 mg, about 16 mg, about 17 mg, about 18 mg, about 19 mg, about 20 mg, about 21 mg, about 22 mg, about 23 mg, about 24 mg, about 25 mg, about 26 mg, about 27 mg, about 28 mg, about 29 mg, about 30 mg, about 31 mg, about 32 mg, about 33 mg, about 34 mg, about 35 mg, about 36 mg, about 37 mg, about 38 mg, about 39 mg, about 40 mg, about 41 mg, about 42 mg, about 43 mg, about 44 mg, about 45 mg, about46 mg, about 47 mg, about 48 mg, about 49 mg, about 50 mg, about 51 mg, about 52 mg, about 53 mg, about 54 mg, about 55 mg, about 56 mg, about 57 mg, about 58 mg, about 59 mg, about 60 mg, about 61 mg, about 62 mg, about 63 mg, about 64 mg, about 65 mg, about 66 mg, about 67 mg, about 68 mg, about 69 mg, about 70 mg, about 71 mg, about 72 mg, about 73 mg, about 74 mg, about 75 mg, about 76 mg, about 77 mg, about 78 mg, about 79 mg, about 80 mg, about 81 mg, about 82 mg, about 83 mg, about 84 mg, about 85 mg, about 86 mg, about 87 mg, about 88 mg, about 89 mg, about 90 mg, about 91 mg, about 92 mg, about 93 mg, about 94 mg, about 95 mg, about 96 mg, about 97 mg, about 98 mg, about 99 mg, or about 100 mg.

[0064] A pharmaceutical composition can comprise one or more lubricants. Non-limiting examples of lubricants can include magnesium stearate, stearic acid, calcium stearate, sodium stearyl fumarate, polyethylene glycol, silicone dioxide, talc, or beeswax. In some embodiments, a lubricant can comprise magnesium stearate.

[0065] In some embodiments, a lubricant (e.g., magnesium stearate) is present in a formulation in an amount of from about 0. 1 mg to about 100 mg, from about 10 mg to about 100 mg, from about 10 mg to about 80 mg, from about 20 mg to about 100 mg, from about 20 mg to about 60 mg, from about 20 mg to about 40 mg, from about 1 mg to about 20 mg, from about 40 mg to about 50 mg, from about 0. 1 mg to about 1 mg, from about 0.5 mg to about 1 mg, from about 0.5 mg to about 10 mg, from about 0. 1 mg to about 5 mg, from about 5 mg to about 10 mg, from about 10 mg to about 15 mg, from about 15 mg to about 20 mg, from about 20 mg to about 25 mg, from about 25 mg to about 30 mg, from about 30 mg to about 35 mg, from about 35 mg to about 40 mg, from about 40 mg to about 45 mg, about 45 mg to about 50 mg, from about 50 mg to about 55 mg, from about 55 mg to about 60 mg, from about 60 mg to about 65 mg, from about 65 mg to about 70 mg, from about 70 mg to about 75 mg, about 75 mg to about 80 mg, from about 80 mg to about 85 mg, from about 85 mg to about 90 mg, from about 90 mg to about 95 mg, or from about 95 mg to about 100 mg.

[0066] In some embodiments, a lubricant (e.g., magnesium stearate) is present in a formulation in an amount of about 1 mg, about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, about 10 mg, about 11 mg, about 12 mg, about 13 mg, about 14 mg, about 15 mg, about 16 mg, about 17 mg, about 18 mg, about 19 mg, about 20 mg, about 21 mg, about 22 mg, about 23 mg, about 24 mg, about 25 mg, about 26 mg, about 27 mg, about 28 mg, about 29 mg, about 30 mg, about 31 mg, about 32 mg, about 33 mg, about 34 mg, about 35 mg, about 36 mg, about 37 mg, about 38 mg, about 39 mg, about 40 mg, about 41 mg, about 42 mg, about 43 mg, about 44 mg, about 45 mg, about 46 mg, about 47 mg, about 48 mg, about 49 mg, about 50 mg, about 51 mg, about 52 mg, about 53 mg, about 54 mg, about 55 mg, about 56 mg, about 57 mg, about 58 mg, about 59 mg, about 60 mg, about 61 mg, about 62 mg, about 63 mg, about 64 mg, about 65 mg, about 66 mg, about 67 mg, about 68 mg, about 69 mg, about 70 mg, about 71 mg, about 72 mg, about 73 mg, about 74 mg, about 75 mg, about 76 mg, about 77 mg, about 78 mg, about 79 mg, about 80 mg, about 81 mg, about 82 mg, about 83 mg, about 84 mg, about 85 mg, about 86 mg, about 87 mg, about 88 mg, about 89 mg, about 90 mg, about 91 mg, about 92 mg, about 93 mg, about 94 mg, about 95 mg, about 96 mg, about 97 mg, about 98 mg, about 99 mg, or about 100 mg.

[0067] Pharmaceutical formulations can include additional carriers, thickeners, diluents, buffers, preservatives, and surface-active agents in addition to the compounds disclosed herein. An excipient can fdl a role as simple and direct as being an inert fdler, or an excipient as used herein can be part of a pH stabilizing system or coating to insure delivery of theingredients safely to the stomach or other desired target.

[0068] A pharmaceutical composition comprising fasudil or a pharmaceutically-acceptable salt thereof can be sublingual formulation. The formulation provides sufficient solubility for fasudil or a pharmaceutically acceptable salt thereof to be incorporated into the sublingual formulation at therapeutically-effective doses and sublingually delivered. The formulation can be an oral disintegrating formulation of fasudil or a pharmaceutically-acceptable salt thereof.

[0069] Pharmaceutical compositions can be provided in the form of a rapid release formulation, in the form of an extended-release formulation, or in the form of an intermediate release formulation. A rapid release form can provide an immediate release. An extended- release formulation can provide a controlled release or a sustained delayed release.

[0070] For buccal or sublingual administration, the compositions can be tablets, lozenges, or gels. For example, a composition can be in a solid dosage form.

[0071] A solid dosage form is a pharmaceutically acceptable composition that is safe for oral administration to subjects (e.g., humans), in which case all excipients in the dosage form are to be used in the oral formulation. A solid dosage form can be an immediate release tablet, an immediate release capsule, a controlled release tablet, a controlled release capsule, a fastdissolving form, a chewable form, a sachet, etc. In some embodiments, the solid dosage form is a tablet.

[0072] A solid dosage form tablet can comprise a single-layer tablet. Alternatively, a solid dosage form tablet can comprise a double-layer tablet. A solid dosage form can comprise a round tablet, a square tablet, a rectangular tablet, a circular tablet, a spherical tablet, an ovoid tablet, a triangular tablet, or a polygonal tablet. A solid dosage form tablet can be convex on one side. A solid dosage form tablet can be convex on two sides (e.g., biconvex).Alternatively, a solid dosage form tablet can be concave one side. A solid dosage form tablet can be concave on two sides (e.g., biconcave).

[0073] A solid dosage form tablet can have a diameter from about 1 mm to about 15 mm, from about 1 mm to about 2 mm, from about 2 mm to about 3 mm, from about 3 mm to about 4 mm, from about 4 mm to about 5 mm, from about 5 mm to about 6 mm, from about 6 mm to about 7 mm, from about 7 mm to about 8 mm, from about 8 mm to about 9 mm, from about 9 mm to about 10 mm, from about 10 mm to about 11 mm, from about 11 mm to about 12 mm, from about 12 mm to about 13 mm, from about 13 mm to about 14 mm, from about 14 mm to about 15 mm, from about 1 mm to about 3 mm, from about 2 mm to about 4 mm, from about 3 mm to about 5 mm, from about 4 mm to about 6 mm, from about 5 mm to about7 mm, from about 6 mm to about 8 mm, from about 7 mm to about 9 mm, from about 8 mm to about 10 mm, from about 9 mm to about 11 mm, from about 10 mm to about 12 mm, from about 11 mm to about 13 mm, from about 12 mm to about 14 mm, or from about 13 mm to about 15 mm. In some examples, a solid dosage form tablet can have a diameter from about 7 mm to about 9 mm. In some examples, a dosage form tablet can have a diameter from about 8 mm to about 8.2 mm.

[0074] A solid dosage form tablet can have a thickness from about from about 1 mm to about 10 mm, from about 1 mm to about 2 mm, from about 1.5 mm to about 2.5 mm, from about 2 mm to about 3 mm, from about 2.5 mm to about 3.5 mm, from about 3 mm to about 4 mm, from about 3.5 mm to about 4.5 mm, from about 4 mm to about 5 mm, from about 4.5 mm to about 5.5 mm, from about 5 mm to about 6 mm, from about 5.5 mm to about 6.5 mm, from about 6 mm to about 7 mm, from about 6.5 mm to about 7.5 mm, from about 7 mm to about 8 mm, from about 7.5 mm to about 8.5 mm, from about 8 mm to about 9 mm, from about 8.5 mm to about 9.5 mm, or from about 9 mm to about 10 mm. In some examples, a solid dosage form tablet can have a thickness from about 3.5 mm to about 4.5 mm. In some examples, a dosage form tablet can have a thickness from about 3.8 mm to about 4 mm.

[0075] Dissolution tests (e.g., a shake test) can be performed on a solid dosage form to simulate the sublingual cavity. A solid dosage form can be subjected to a shake test in which the solid dosage form is placed in water and the shaken to disintegrate the solid dosage form in the water. A solid dosage form can disintegrate into fine particles during a shake test in less than about 20 seconds, less than about 25 seconds, less than about 30 seconds, less than about 35 seconds, less than about 40 seconds, less than about 45 seconds, less than about 50 seconds, less than about 55 seconds, less than about 60 seconds, less than about 65 seconds, less than about 70 seconds, less than about 75 seconds, less than about 80 seconds, less than about 85 seconds, or less than about 90 seconds. In some embodiments, a solid dosage form can disintegrate into fine particles during a shake test in less than about 60 seconds. In some embodiments, a solid dosage form can disintegrate into fine particles during a shake test in less than about 40 seconds. In some embodiments, a solid dosage form can disintegrate into fine particles during a shake test in about 30± 2 seconds.

[0076] Thermal analysis can be performed on a solid dosage form to determine stability of the solid dosage form. A dosage form (e.g., a sublingual dosage form) can be heated. The resulting thermograph from the thermal analysis can show an endothermic peak from about 145-170 °C. Alternatively, the resulting thermograph from the thermal analysis can show an endothermic peak from about 156-161 °C.

[0077] In practicing the methods of treatment or use provided herein, therapeutically- effective amounts of the compounds described herein are administered in pharmaceutical compositions to a subject having a disease or condition to be treated. In some embodiments, the subject is a mammal such as a human. A therapeutically-effective amount can vary widely depending on the severity of the disease, the age and relative health of the subject, the potency of the compounds used, and other factors.

[0078] A composition of the invention can be, for example, an immediate release form or a controlled release formulation. An immediate release formulation can be formulated to allow the compounds to act rapidly. Non-limiting examples of immediate release formulations include readily dissolvable formulations. A controlled release formulation can be a pharmaceutical formulation that has been adapted such that drug release rates and drug release profiles can be matched to physiological and chronotherapeutic requirements or, alternatively, has been formulated to effect release of a drug at a programmed rate. Nonlimiting examples of controlled release formulations include granules, delayed release granules, hydrogels (e.g., of synthetic or natural origin), other gelling agents (e.g., gelforming dietary fibers), matrix-based formulations (e.g., formulations comprising a polymeric material having at least one active ingredient dispersed through), granules within a matrix, polymeric mixtures, and granular masses.

[0079] In some, a controlled release formulation is a delayed release form. A delayed release form can be formulated to delay a compound's action for an extended period of time. A delayed release form can be formulated to delay the release of an effective dose of one or more compounds, for example, for about 4, about 8, about 12, about 16, or about 24 hours.

[0080] A controlled release formulation can be a sustained release form. A sustained release form can be formulated to sustain, for example, the compound's action over an extended period of time. A sustained release form can be formulated to provide an effective dose of any compound described herein (e.g., provide a physiologically-effective blood profile) over about 4, about 8, about 12, about 16 or about 24 hours.

[0081] Non-limiting examples of pharmaceutically-acceptable excipients can be found, for example, in Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa. Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa. 1975; Liberman, H. A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y., 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins 1999), each of which is incorporated by reference in its entirety.

[0082] The disclosed methods include administration of a fasudil or a pharmaceutically- acceptable salt thereof, in combination with a pharmaceutically-acceptable carrier. The carrier can be selected to minimize any degradation of the active ingredient and to minimize any adverse side effects in the subject.

[0083] The fasudil or a pharmaceutically-acceptable salt thereof as described herein can be formulated into pharmaceutical compositions composed of one or more pharmaceutically- acceptable carriers. See e.g., Remington's Pharmaceutical Sciences, latest edition, by E.W. Martin Mack Pub. Co., Easton, Pa., which discloses carriers and methods of preparing pharmaceutical compositions that can be used in conjunction with the preparation of formulations of the compound described herein and which is incorporated by reference herein.

[0084] The disclosed methods relate to sublingual administration of fasudil or a pharmaceutically-acceptable salt thereof as part of a pharmaceutical composition. In some embodiments, a method of treating a disease of a subject by administering the sublingual formulation is provided. The method comprises providing a sublingual formulation as described herein having an effective amount of fasudil or a pharmaceutically-acceptable salt, solvate, anomer, hydrate, or prodrug thereof, and administering the formulation to a subject to treat the diseased state.Pharmaceutical Compositions

[0085] Pharmaceutical compositions containing the compounds described herein can be administered for prophylactic or therapeutic treatments. In therapeutic applications, the compositions can be administered to a subject already suffering from a disease or condition, in an amount sufficient to cure or at least partially arrest the symptoms of the disease or condition, or to cure, heal, improve, reduce, lessen, or ameliorate the disease or condition. Compounds can also be administered to lessen or reduce a likelihood of developing, contracting, or worsening a condition. Amounts effective for this use can vary based on the severity and course of the disease or condition, previous therapy, the subject's health status, weight, response to the drugs, and the judgment of the treating physician.

[0086] Compounds and compositions of the invention can be packaged as a kit. In some embodiments, the invention provides a kit comprising a compound disclosed herein, or a pharmaceutically-acceptable salt thereof, and written instructions on use of the kit in the treatment of a condition described herein. In some embodiments, the invention provides a kit comprising a compound disclosed herein, or a pharmaceutically-acceptable salt thereof, and written instructions on use of the kit in the treatment of a condition described herein.

[0087] The compounds described herein can be administered before, during, or after the occurrence of a disease or condition, and the timing of administering the composition containing a compound can vary. For example, the compounds can be used as a prophylactic and can be administered continuously to subjects with a propensity to conditions or diseases in order to lessen or reduce a likelihood of the occurrence of the disease or condition. The compounds and compositions can be administered to a subject during or as soon as possible after the onset of the symptoms. The administration of the compounds can be initiated within the first 48 hours of the onset of the symptoms, within the first 24 hours of the onset of the symptoms, within the first 6 hours of the onset of the symptoms, or within 3 hours of the onset of the symptoms. The initial administration can be via any route practical, such as by any route described herein using any formulation described herein.

[0088] A compound can be administered as soon as is practical after the onset of a disease or condition is detected or suspected, and for a length of time necessary for the treatment of the disease, such as, for example, from about 1 month to about 3 months. In some embodiments, the length of time a compound can be administered can be about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 6 days, about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 1 month, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 2 months, about 9 weeks, about 10 weeks, about 11 weeks, about 12 weeks, about 3 months, about 13 weeks, about 14 weeks, about 15 weeks, about 16 weeks, about 4 months, about 17 weeks, about 18 weeks, about 19 weeks, about 20 weeks, about 5 months, about 21 weeks, about 22 weeks, about 23 weeks, about 24 weeks, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, about 1 year, about 13 months, about 14 months, about 15 months, about 16 months, about 17 months, about 18 months, about 19 months, about 20 months, about 21 months, about 22 months about 23 months, about 2 years, about 2.5 years, about 3 years, about 3.5 years, about 4 years, about 4.5 years, about 5 years, about 6 years, about 7 years, about 8 years, about 9 years, or about 10 years. The length of treatment can vary for each subject.

[0089] Pharmaceutical compositions described herein can be in unit dosage forms suitable for single administration of precise dosages. In unit dosage form, the formulation is divided into unit doses containing appropriate quantities of one or more compounds. The unit dosage can be in the form of a package containing discrete quantities of the formulation. Nonlimiting examples are packaged injectables, vials, or ampoules. Aqueous suspension compositions can be packaged in single-dose non-reclosable containers. Multiple-dose reclosable containers can be used, for example, in combination with or without apreservative. Formulations for parenteral injection can be presented in unit dosage form, for example, in ampoules, or in multi-dose containers with a preservative.

[0090] A fasudil compound described herein can be present in a composition in a range of from about 1 mg to about 5 mg, from about 5 mg to about 10 mg, from about 10 mg to about 15 mg, from about 15 mg to about 20 mg, from about 20 mg to about 25 mg, from about 25 mg to about 30 mg, from about 30 mg to about 35 mg, from about 35 mg to about 40 mg, from about 40 mg to about 45 mg, from about 45 mg to about 50 mg, from about 50 mg to about 55 mg, from about 55 mg to about 60 mg, from about 60 mg to about 65 mg, from about 65 mg to about 70 mg, from about 70 mg to about 75 mg, from about 75 mg to about 80 mg, from about 80 mg to about 85 mg, from about 85 mg to about 90 mg, from about 90 mg to about 95 mg, from about 95 mg to about 100 mg, from about 100 mg to about 125 mg, from about 125 mg to about 150 mg, from about 150 mg to about 175 mg, from about 175 mg to about 200 mg, from about 200 mg to about 225 mg, from about 225 mg to about 250 mg, or from about 250 mg to about 300 mg.

[0091] A fasudil compound described herein can be present in a composition in an amount of about 1 mg, about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, about 100 mg, about 125 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about 250 mg, or about 300 mg.

[0092] A sublingual formulation of fasudil can achieve a lower therapeutic dose than that of orally administered fasudil. A sublingual formulation of fasudil can achieve a therapeutic dose that is about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, or about 150 mg less than that of an amount of orally administered fasudil that has a similar or same therapeutic effect.

[0093] A sublingual formulation of fasudil can produce a rapid therapeutic onset of action. A sublingual formulation of fasudil can produce a therapeutic onset of action within about 10 seconds, about 15 seconds, about 20 seconds, about 25 seconds, about 30 seconds, about 35 seconds, about 40 seconds, about 45 seconds, about 50 seconds, about 55 seconds, about 1 minute, about 2 minutes, about 3 minutes, about 4 minutes, about 5 minutes, about 6 minutes, about 7 minutes, about 8 minutes, about 9 minutes, about 10 minutes, about 11 minutes, about 12 minutes, about 13 minutes, about 14 minutes, about 15 minutes, about 16 minutes, about 17 minutes, about 18 minutes, about 19 minutes, about 20 minutes, about 21 minutes,about 22 minutes, about 23 minutes, about 24 minutes, about 25 minutes, about 26 minutes, about 27 minutes, about 28 minutes, about 29 minutes, about 30 minutes, about 35 minutes, about 40 minutes, about 45 minutes, about 50 minutes, about 55 minutes, or about 1 hour.

[0094] A sublingual formulation of fasudil can produce a therapeutic onset of action faster than that of orally administered fasudil. A sublingual formulation of fasudil can produce a therapeutic onset of action at least about 1 minute, at least about 2 minutes, at least about 3 minutes, at least about 4 minutes, at least about 5 minutes, at least about 6 minutes, at least about 7 minutes, at least about 8 minutes, at least about 9 minutes, at least about 10 minutes, at least about 11 minutes, at least about 12 minutes, at least about 13 minutes, at least about 14 minutes, at least about 15 minutes, at least about 16 minutes, at least about 17 minutes, at least about 18 minutes, at least about 19 minutes, at least about 20 minutes, at least about 21 minutes, at least about 22 minutes, at least about 23 minutes, at least about 24 minutes, at least about 25 minutes, at least about 26 minutes, at least about 27 minutes, at least about 28 minutes, at least about 29 minutes, at least about 30 minutes, at least about 35 minutes, at least about 40 minutes, at least about 45 minutes, at least about 50 minutes, at least about 55 minutes, at least about 1 hour, at least about 2 hours, at least about 3 hours, at least about 4 hours, at least about 5 hours, or at least about 6 hours faster than that of orally administered fasudil.

[0095] A sublingual formulation of fasudil can reduce side effects of fasudil. A sublingual formulation of fasudil can reduce the total drug load necessary to result in a therapeutic effect. In some embodiments, a lower sublingual dosage formulation of fasudil can deliver similar therapeutic effects compared to a higher oral dosage formulation of fasudil or enhanced therapeutic effects as compared to a higher oral dosage formulation. In some embodiments, a sublingual dosage formulation of fasudil can reduce side effects of fasudil as compared to an oral formulation of fasudil.Treatment of Subjects with Neurodegenerative Disease

[0096] The invention discloses methods for treating a subject afflicted a neurodegenerative disease. A neurodegenerative disease is a progressive disease in which cells of the central nervous system stop working and / or die. Non-limiting examples of neurodegenerative diseases include Alzheimer’s disease, Parkinson’s disease, prion disease, Amyotrophic lateral sclerosis (ALS), motor neuron disease, Huntington’s disease, dementia, spinal muscular atrophy, and spinocerebellar ataxia.

[0097] Amyotrophic lateral sclerosis (ALS), also known as Lou Gehrig’s disease, is a neurodegenerative disease that results in the progressive loss of motor neurons that controlvoluntary muscles. Different types of ALS can be classified by the types of motor neurons that are affected - upper motor neurons (e.g., in the motor cortex of the brain) or lower motor neurons (e.g., neurons of the spinal cord).

[0098] Dementia is a disorder characterized by the loss of cognitive function (e.g., thinking, remembering, reasoning) to such an extent that interferes with daily life. One form of dementia is Alzheimer’s disease. Alzheimer’s disease is a neurodegenerative disease that interferes with cognitive and functional impairment (e.g., memory impairment).

[0099] Parkinson’s disease is a neurodegenerative disorder characterized by unintended or uncontrollable movements such as shaking, stiffness, or difficulty with balance and coordination. Parkinson’s disease can be caused by falling dopamine levels, which can be caused by nerve cell damage.

[0100] The subject can be a human. Treatment can include treating a human in a clinical trial. A treatment can comprise administering to a subject a pharmaceutical composition comprising one or more of fasudil or a pharmaceutically-acceptable salt thereof, as described throughout the disclosure.

[0101] In some embodiments, the invention provides fasudil or a pharmaceutically- acceptable salt thereof for use in treatment of a neurodegenerative disease (e.g., ALS, dementia, Parkinson’s disease, Alzheimer’s disease). In some embodiments, the invention provides fasudil or a pharmaceutically-acceptable salt thereof for use in the manufacture of a medicament for the treatment of a neurodegenerative disease (e.g., ALS, dementia, Parkinson’s disease, Alzheimer’s disease).

[0102] Treatment of a neurodegenerative disease (e.g., ALS, dementia, Parkinson’s disease, Alzheimer’s disease) can result in reducing muscle weakness by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 100%.

[0103] Treatment of a neurodegenerative disease (e.g., ALS, dementia, Parkinson’s disease, Alzheimer’s disease) can result in reducing motor neuron loss by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 100%.

[0104] Non-limiting examples of possible subjects for administration include the following.Subjects can be humans, non-human primates such as chimpanzees, and other apes and monkey species; farm animals such as cattle, horses, sheep, goats, and swine; domestic animals such as rabbits, dogs, and cats; and laboratory animals including rats, mice, and guinea pigs. A subject can be of any age. Subjects can be, for example, elderly adults, adults, adolescents, pre-adolescents, children, toddlers, and infants.EMBODIMENTS

[0105] Embodiment 1. A pharmaceutical composition comprising fasudil or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition is a sublingual dosage form.

[0106] Embodiment 2. The pharmaceutical composition of embodiment 1, wherein the pharmaceutical composition further comprises a pharmaceutically-acceptable excipient.

[0107] Embodiment 3. The pharmaceutical composition of embodiment 1, wherein the pharmaceutical composition further comprises cellulose.

[0108] Embodiment 4. The pharmaceutical composition of embodiment 1, wherein the pharmaceutical composition further comprises mannitol.

[0109] Embodiment 5. The pharmaceutical composition of embodiment 1, wherein the pharmaceutical composition further comprises sodium starch glycolate.

[0110] Embodiment 6. The pharmaceutical composition of embodiment 1, wherein the pharmaceutical composition further comprises magnesium stearate.

[0111] Embodiment 7. The pharmaceutical composition of embodiment 1, wherein the fasudil or the pharmaceutically-acceptable salt thereof is present in the pharmaceutical composition in a concentration of about 50 nM to about 300 nM.

[0112] Embodiment 8. The pharmaceutical composition of embodiment 1, wherein the fasudil or the pharmaceutically acceptable salt thereof is present in the pharmaceutical composition in a concentration of at least 1 pM.

[0113] Embodiment 9. The pharmaceutical composition of embodiment 1, wherein the pharmaceutical composition comprises the pharmaceutically-acceptable salt, and the pharmaceutically acceptable salt is a hydrochloride salt.

[0114] Embodiment 10. The pharmaceutical composition of embodiment 1, wherein the fasudil or the pharmaceutically acceptable salt thereof is present in the pharmaceutical composition in an amount of about 20 mg to about 60 mg.

[0115] Embodiment 11. The pharmaceutical composition of embodiment 1, wherein the fasudil or the pharmaceutically acceptable salt thereof is present in the pharmaceutical composition in an amount of about 40 mg to about 50 mg.

[0116] Embodiment 12. The pharmaceutical composition of embodiment 1, wherein the fasudil or the pharmaceutically acceptable salt thereof is present in the pharmaceutical composition in an amount of about 45 mg.

[0117] Embodiment 13. The pharmaceutical composition of embodiment 1, further comprising a filler, a binder, a superdisintegrant, and a lubricant.

[0118] Embodiment 14. The pharmaceutical composition of embodiment 1, further comprising microcrystalline cellulose, mannitol, sodium starch glycolate, and magnesium stearate.

[0119] Embodiment 15. The pharmaceutical composition of embodiment 1, further comprising: a) microcrystalline cellulose in an amount of about 20 mg to about 40 mg; b) mannitol in an amount of a about 20 mg to about 100 mg; c) sodium starch glycolate in an amount of about 1 mg to about 20 mg; and d) magnesium stearate in an amount of about 0.5 mg to about 10 mg.

[0120] Embodiment 16. The pharmaceutical composition of embodiment 1, further comprising: a) microcrystalline cellulose in an amount of about 33 mg; b) mannitol in an amount of a about 60 mg; c) sodium starch glycolate in an amount of about 9 mg; and d) magnesium stearate in an amount of about 3 mg.

[0121] Embodiment 17. The pharmaceutical composition of embodiment 1, wherein the sublingual dosage form is a tablet.

[0122] Embodiment 18. The pharmaceutical composition of embodiment 1, wherein the sublingual dosage form is a tablet that is round and biconcave.

[0123] Embodiment 19. The pharmaceutical composition of embodiment 1, wherein the sublingual dosage form is a tablet that has a diameter from about 7 mm to about 9 mm and a thickness from about 3.5 mm to about 4.5 mm.

[0124] Embodiment 20. The pharmaceutical composition of embodiment 1, wherein the sublingual dosage form is a tablet that has a diameter from about 8 mm to about 8.2 mm and a thickness from about 3.8 mm to about 4 mm.

[0125] Embodiment 21. The pharmaceutical composition of embodiment 1, wherein the sublingual dosage form is solid, wherein if the sublingual dosage form is subjected to a shake test in which the sublingual dosage form is placed in a glass beaker containing 2 mL of water and the beaker is manually shaken to disintegrate the sublingual dosage form in the water, then the time needed for disintegration of the sublingual dosage form into fine particles is less than sixty seconds.

[0126] Embodiment 22. The pharmaceutical composition of embodiment 1, wherein the sublingual dosage form is solid, wherein if the sublingual dosage form is subjected to a shake test in which the sublingual dosage form is placed in a glass beaker containing 2 mL of water and the beaker is manually shaken to disintegrate the sublingual dosage form in the water, then the time needed for disintegration of the sublingual dosage form into fine particles is less than forty seconds.

[0127] Embodiment 23. The pharmaceutical composition of embodiment 1, wherein the sublingual dosage form is solid, wherein if the sublingual dosage form is subjected to a shake test in which the sublingual dosage form is placed in a glass beaker containing 2 mL of water and the beaker is manually shaken to disintegrate the sublingual dosage form in the water, then the time needed for disintegration of the sublingual dosage form into fine particles is 30 ± 2 seconds.

[0128] Embodiment 24. The pharmaceutical composition of embodiment 1, wherein if a sample of about 8 mg to about 13 mg of the sublingual dosage form is subjected to differential thermal analysis by placing the sample in a platinum pan of 100 pL capacity, and the sample is heated from 25 °C to 600 °C at a rate of 10 °C / min-l under air flow, then a resulting thermograph shows an endothermic peak at 145-170 °C and does not show an endothermic peak at 210-235 °C.

[0129] Embodiment 25. The pharmaceutical composition of embodiment 1, wherein if a sample of about 8 mg to about 13 mg of the sublingual dosage form is subjected to differential thermal analysis by placing the sample in a platinum pan of 100 pL capacity, and the sample is heated from 25 °C to 600 °C at a rate of 10 °C / min-l under air flow, then a resulting thermograph shows an endothermic peak at 156-161 °C and does not show an endothermic peak at 218-223 °C.

[0130] Embodiment 26. The pharmaceutical composition of embodiment 1, wherein the sublingual dosage form is solid, wherein if the sublingual dosage form is subjected to a dissolution study in which the sublingual dosage form is dipped and held in a medium of 2 mL of distilled water at 25 °C in a 15 mL glass funnel for sixty seconds, at which point themedium is drawn by vacuum through a 0.45 gm membrane into a collection tube, after which the medium is analyzed by HPLC, then a percent dissolution of fasudil or the pharmaceutically acceptable salt thereof of 80% - 100% is obtained.

[0131] Embodiment 27. The pharmaceutical composition of embodiment 1, wherein the sublingual dosage form is solid, wherein if the sublingual dosage form is subjected to a dissolution study in which the sublingual dosage form is dipped and held in a medium of 2 mL of distilled water at 25 °C in a 15 mL glass funnel for sixty seconds, at which point the medium is drawn by vacuum through a 0.45 pm membrane into a collection tube, after which the medium is analyzed by HPLC, then a percent dissolution of fasudil or the pharmaceutically acceptable salt thereof of 85% - 95% is obtained.

[0132] Embodiment 28. The pharmaceutical composition of embodiment 1, wherein the sublingual dosage form is solid, wherein if the sublingual dosage form is subjected to a dissolution study in which the sublingual dosage form is dipped and held in a medium of 2 mL of distilled water at 25 °C in a 15 mL glass funnel for sixty seconds, at which point the medium is drawn by vacuum through a 0.45 pm membrane into a collection tube, after which the medium is analyzed by HPLC, then a percent dissolution of fasudil or the pharmaceutically acceptable salt thereof of 89.6 ± 4.0% is obtained.

[0133] Embodiment 29. A pharmaceutical composition comprising fasudil or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition is a solid dosage form, wherein if the solid dosage form is subjected to a shake test in which the solid dosage form is placed in a glass beaker containing 2 mL of water and the beaker is manually shaken to disintegrate the solid dosage form in the water, then the time needed for disintegration of the solid dosage form into fine particles is less than sixty seconds.

[0134] Embodiment 30. The pharmaceutical composition of embodiment 29, wherein the time needed for disintegration of the solid dosage form into fine particles is less than forty seconds.

[0135] Embodiment 31. The pharmaceutical composition of embodiment 29, wherein the time needed for disintegration of the solid dosage form into fine particles is 30 ± 2 seconds.

[0136] Embodiment 32. The pharmaceutical composition of embodiment 29, wherein the pharmaceutical composition further comprises a pharmaceutically-acceptable excipient.

[0137] Embodiment 33. The pharmaceutical composition of embodiment 29, wherein the pharmaceutical composition further comprises cellulose.

[0138] Embodiment 34. The pharmaceutical composition of embodiment 29, wherein the pharmaceutical composition further comprises mannitol.

[0139] Embodiment 35. The pharmaceutical composition of embodiment 29, wherein the pharmaceutical composition further comprises sodium starch glycolate.

[0140] Embodiment 36. The pharmaceutical composition of embodiment 29, wherein the pharmaceutical composition further comprises magnesium stearate.

[0141] Embodiment 37. The pharmaceutical composition of embodiment 29, wherein the fasudil or the pharmaceutically-acceptable salt thereof is present in the pharmaceutical composition in a concentration of about 50 nM to about 300 nM.

[0142] Embodiment 38. The pharmaceutical composition of embodiment 29, wherein the fasudil or the pharmaceutically acceptable salt thereof is present in the pharmaceutical composition in a concentration of at least 1 pM.

[0143] Embodiment 39. The pharmaceutical composition of embodiment 29, wherein the pharmaceutical composition comprises the pharmaceutically-acceptable salt, and the pharmaceutically acceptable salt is a hydrochloride salt.

[0144] Embodiment 40. The pharmaceutical composition of embodiment 29, wherein the fasudil or the pharmaceutically acceptable salt thereof is present in the pharmaceutical composition in an amount of about 20 mg to about 60 mg.

[0145] Embodiment 41. The pharmaceutical composition of embodiment 29, wherein the fasudil or the pharmaceutically acceptable salt thereof is present in the pharmaceutical composition in an amount of about 40 mg to about 50 mg.

[0146] Embodiment 42. The pharmaceutical composition of embodiment 29, wherein the fasudil or the pharmaceutically acceptable salt thereof is present in the pharmaceutical composition in an amount of about 45 mg.

[0147] Embodiment 43. The pharmaceutical composition of embodiment 29, further comprising a filler, a binder, a superdisintegrant, and a lubricant.

[0148] Embodiment 44. The pharmaceutical composition of embodiment 29, further comprising microcrystalline cellulose, mannitol, sodium starch glycolate, and magnesium stearate.

[0149] Embodiment 45. The pharmaceutical composition of embodiment 29, further comprising: a) microcrystalline cellulose in an amount of about 20 mg to about 40 mg; b) mannitol in an amount of a about 20 mg to about 100 mg; c) sodium starch glycolate in an amount of about 1 mg to about 20 mg; andd) magnesium stearate in an amount of about 0.5 mg to about 10 mg.

[0150] Embodiment 46. The pharmaceutical composition of embodiment 29, further comprising: a) microcrystalline cellulose in an amount of about 33 mg; b) mannitol in an amount of a about 60 mg; c) sodium starch glycolate in an amount of about 9 mg; and d) magnesium stearate in an amount of about 3 mg.

[0151] Embodiment 47. The pharmaceutical composition of embodiment 29, wherein the solid dosage form is a tablet.

[0152] Embodiment 48. The pharmaceutical composition of embodiment 29, wherein the solid dosage form is a tablet that is round and biconcave.

[0153] Embodiment 49. The pharmaceutical composition of embodiment 29, wherein the solid dosage form is a tablet that has a diameter from about 7 mm to about 9 mm and a thickness from about 3.5 mm to about 4.5 mm.

[0154] Embodiment 50. The pharmaceutical composition of embodiment 29, wherein the solid dosage form is a tablet that has a diameter from about 8 mm to about 8.2 mm and a thickness from about 3.8 mm to about 4 mm.

[0155] Embodiment 51. A pharmaceutical composition comprising fasudil or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition is a solid dosage form, wherein if a sample of about 8 mg to about 13 mg of the solid dosage form is subjected to differential thermal analysis by placing the sample in a platinum pan of 100 pL capacity, and the sample is heated from 25 °C to 600 °C at a rate of 10 °C / min-l under air flow, then a resulting thermograph shows an endothermic peak at 145-170 °C and does not show an endothermic peak at 210-235 °C.

[0156] Embodiment 52. The pharmaceutical composition of embodiment 51, wherein the resulting thermograph shows an endothermic peak at 156-161 °C and does not show an endothermic peak at 218-223 °C.

[0157] Embodiment 53. The pharmaceutical composition of embodiment 51, wherein the pharmaceutical composition further comprises a pharmaceutically-acceptable excipient.

[0158] Embodiment 54. The pharmaceutical composition of embodiment 51, wherein the pharmaceutical composition further comprises cellulose.

[0159] Embodiment 55. The pharmaceutical composition of embodiment 51, wherein the pharmaceutical composition further comprises mannitol.

[0160] Embodiment 56. The pharmaceutical composition of embodiment 51, wherein the pharmaceutical composition further comprises sodium starch glycolate.

[0161] Embodiment 57. The pharmaceutical composition of embodiment 51, wherein the pharmaceutical composition further comprises magnesium stearate.

[0162] Embodiment 58. The pharmaceutical composition of embodiment 51, wherein the fasudil or the pharmaceutically-acceptable salt thereof is present in the pharmaceutical composition in a concentration of about 50 nM to about 300 nM.

[0163] Embodiment 59. The pharmaceutical composition of embodiment 51, wherein the fasudil or the pharmaceutically acceptable salt thereof is present in the pharmaceutical composition in a concentration of at least 1 pM.

[0164] Embodiment 60. The pharmaceutical composition of embodiment 51, wherein the pharmaceutical composition comprises the pharmaceutically-acceptable salt, and the pharmaceutically acceptable salt is a hydrochloride salt.

[0165] Embodiment 61. The pharmaceutical composition of embodiment 51, wherein the fasudil or the pharmaceutically acceptable salt thereof is present in the pharmaceutical composition in an amount of about 20 mg to about 60 mg.

[0166] Embodiment 62. The pharmaceutical composition of embodiment 51, wherein the fasudil or the pharmaceutically acceptable salt thereof is present in the pharmaceutical composition in an amount of about 40 mg to about 50 mg.

[0167] Embodiment 63. The pharmaceutical composition of embodiment 51, wherein the fasudil or the pharmaceutically acceptable salt thereof is present in the pharmaceutical composition in an amount of about 45 mg.

[0168] Embodiment 64. The pharmaceutical composition of embodiment 51, further comprising a filler, a binder, a superdisintegrant, and a lubricant.

[0169] Embodiment 65. The pharmaceutical composition of embodiment 51, further comprising microcrystalline cellulose, mannitol, sodium starch glycolate, and magnesium stearate.

[0170] Embodiment 66. The pharmaceutical composition of embodiment 51, further comprising: a) microcrystalline cellulose in an amount of about 20 mg to about 40 mg; b) mannitol in an amount of a about 20 mg to about 100 mg; c) sodium starch glycolate in an amount of about 1 mg to about 20 mg; and d) magnesium stearate in an amount of about 0.5 mg to about 10 mg.

[0171] Embodiment 67. The pharmaceutical composition of embodiment 51, further comprising: a) microcrystalline cellulose in an amount of about 33 mg; b) mannitol in an amount of a about 60 mg; c) sodium starch glycol ate in an amount of about 9 mg; and d) magnesium stearate in an amount of about 3 mg.

[0172] Embodiment 68. The pharmaceutical composition of embodiment 51, wherein the solid dosage form is a tablet.

[0173] Embodiment 69. The pharmaceutical composition of embodiment 51, wherein the solid dosage form is a tablet that is round and biconcave.

[0174] Embodiment 70. The pharmaceutical composition of embodiment 51, wherein the solid dosage form is a tablet that has a diameter from about 7 mm to about 9 mm and a thickness from about 3.5 mm to about 4.5 mm.

[0175] Embodiment 71. The pharmaceutical composition of embodiment 51, wherein the solid dosage form is a tablet that has a diameter from about 8 mm to about 8.2 mm and a thickness from about 3.8 mm to about 4 mm.

[0176] Embodiment 72. A method for treating a condition, the method comprising sublingually administering to a subject in need thereof a therapeutically-effective amount of fasudil or a pharmaceutically acceptable salt thereof.

[0177] Embodiment 73. A method for treating a condition, the method comprising administering to a subject in need thereof a therapeutically-effective amount of the pharmaceutical composition of any one of embodiments 1-71.

[0178] Embodiment 74. The method of any one of embodiments 72 or 73, wherein the condition is a neurodegenerative disease.

[0179] Embodiment 75. The method of any one of embodiments 72-74, wherein the condition is a central nervous system disease.

[0180] Embodiment 76. The method of any one of embodiments 72-75, wherein the condition is an aging condition.

[0181] Embodiment 77. The method of any one of embodiments 72-76, wherein the condition is amyotrophic lateral sclerosis (ALS).

[0182] Embodiment 78. The method of any one of embodiments 72-76, wherein the condition is dementia.

[0183] Embodiment 79. The method of any one of embodiments 72-76, wherein the condition is Alzheimer’s Disease.

[0184] Embodiment 80. The method of any one of embodiments 72-76, wherein the condition is Parkinson’s Disease.

[0185] Embodiment 8E The method of any one of embodiments 72-80, wherein the therapeutically-effective amount is about 20 mg to about 60 mg.

[0186] Embodiment 82. The method of any one of embodiments 72-81, wherein the therapeutically-effective amount is about 40 mg to about 50 mg.

[0187] Embodiment 83. The method of any one of embodiments 72-82, wherein the therapeutically-effective amount is about 45 mg.

[0188] Embodiment 84. The method of any one of embodiments 72-83, wherein treating the condition reduces muscle weakness in the subject by at least 10%.

[0189] Embodiment 85. The method of any one of embodiments 72-83, wherein treating the condition reduces motor neuron loss in the subject by at least 10%.EXAMPLESEXAMPLE 1: General Methods

[0190] Chemical synthesis of fasudil and fasudil hydrochloride

[0191] The starting material for synthesis of fasudil and fasudil hydrochloride was isoquinoline. The chemical synthesis procedure used is shown in FIG. 1. Confirmation of synthesis was determined byJH NMR and IR spectra (FIGs. 2A-2B). Synthesized fasudil was confirmed to have the pharmacokinetic properties shown in Table 1.Table 1: Pharmacokinetic Properties of Fasudil

[0192] Creation of a placebo control

[0193] The placebo solution was prepared by mixing the selected excipients in the same proportions as the tablet formulation. 73 mg of microcrystalline cellulose, 133 mg of mannitol, 20 mg of sodium starch glycolate and 6.6 mg of magnesium stearate were suspended in about 80 mL of mobile phase. The mixture was sonicated for about 5 min. The volume was then completed to 100 mL with the same solvent. About 20 mL of the solution were taken, spun by centrifuge at 8000 G for 10 min, and filtered through 0.45 pm PVDF membrane.

[0194] Creation of fasudil hydrochloride solutions

[0195] A fasudil hydrochloride solution was formed by dissolving! 0 mg fasudil hydrochloride into 8 mL of a mobile phase. The mobile phase used in this experiment was 100 pL of trifluoroacetic acid dissolved in a water / acetonitrile (85:15 v / v) solution. After complete dissolution, the volume was completed to the gauge in a 10 mL volumetric flask to obtain a fasudil hydrochloride concentration of 1 mg / mL stock solution.

[0196] Five standard solutions of FSD hydrochloride were prepared in concentrations between 10 pg / mL and 100 pg / mL by diluting the stock solution in the mobile phase. To evaluate the effect of excipients on the chromatographic separation, five reconstituted solutions were prepared with FSD concentrations ranging from 10 pg / mL to 100 pg / mL by diluting the stock solutions and placebo in the mobile phase in appropriate proportions. Table 2 shows respective volumes of standard solution preparations.Table 2: Fasudil Standard Solutions

[0197] To evaluate the effect of excipients on the chromatographic separation, five reconstituted solutions were prepared with FSD concentrations ranging from 10 pg / mL to 100 pg / mL by diluting the stock solutions and placebo in the mobile phase in appropriate proportions. The volumes of such preparation are reported in Table 3.Table 3: Fasudil Reconstituted Solutions

[0198] In vitro characterization of fasudil

[0199] Fasudil (FSD) hydrochloride (100 pM) inhibited cell spreading, the formation ofstress fibers, and expression of a-SMA with concomitant suppression of cell growth in rat HSCs (hepatic stellate cells) and human HSC-derived TWNT-4 cells. FSD hydrochloride (50-100 pM; 24 hours) inhibited the LPA (lysophosphatidic acid)-induced phosphorylation of ERK1 / 2, JNK, and p38 detected by western blotting in rat HSCs and human HSC-derived TWNT-4 cells. FSD hydrochloride (25-100 pM; 24 hours) suppressed transcription of collagen and TIMP and stimulated transcription of MMP-1 in human HSC-derived TWNT-4 cells. FSD hydrochloride (25-100 pM; 24 hours) increased active MMP-1 and pro-MMP-1 activity in human HSC-derived TWNT-4 cells as determined with a MMP-1 activity assay, increased MMP-1 expression and decreased TIMP-1 expression detected by ELISA.

[0200] RT-PCR experiments also used rat HSCs and human HSC-derived TWNT-4 cells FSD hydrochloride (25-100 pM; 24 hours) reduced the expression of type I collagen, a-SMA, and TIMP-1 normalized to glyceraldehyde-3 -phosphate dehydrogenase (GAPDH) expression. Type I collagen expression was assessed with 5’- AGGGTGAGACAGGCGAACAG-3’ (SEQ ID NO. l)(forward primer) and 5’- CTCTTGAGGTGGCTGGGGCA-3’ (SEQ ID NO. 2) (reverse primer). SMA expression was assessed with 5’-AATGAGATGGCCACTGCCGC-3’ (SEQ ID NO. 3) (forward primer) and 5’-CAGAGTATTTGCGCTCCGGA-3’ (SEQ ID NO. 4) (reverse primer). MMP1 expression was assessed with 5’-GATCATCGGGACAACTCTCCT-3’ (SEQ ID NO. 5) (forward primer) and 5’-TCCGGGTAGAAGGGATTTGTG-3’(SEQ ID NO. 6)(reverse primer).TIMP-1 expression was assessed with 5’-TTCTGCAATTCCGACCTCGT-3’ (SEQ ID NO.7) (forward primer) and 5’-TCCGTCCACAAGCAATGAGT-3’ (SEQ ID NO. 8) (reverse primer).

[0201] Each FSD standard solution was calibrated by the creation of a calibration curve wherein peak area versus FSD concentrations was plotted and a regression coefficient was calculated from the straight-line equation. The FSD concentration was determined from the peak area.

[0202] The concentration of standard solutions was obtained from the following equation: 100where mF is the FSD hydrochloride weight used for the preparation of FSD stock solution in mg, P is the purity of the FSD hydrochloride standard in decimal (e.g., 0.99), V is the volume of the stock solution sample and Vd is the final dilution volume of standard solution (e.g., 5 mL).

[0203] The concentration of FSD hydrochloride in the test solutions were obtained from thefollowing formula:

[0204] where AF is the area of FSD peak in mAU*min, b is the intercept of calibration curve and a is the slope of calibration curve.

[0205] The concentration of FSD in the test solutions were obtained using the following formula:where CFCI is the concentration of FSD hydrochloride in pg / mL, MF is the molecular weight of FSD (i.e. 291.37 g / mol) and MFCI is the molecular weight of FSD hydrochloride (e.g., 327.83 g / mol).

[0206] High Performance Liquid Chromatography (HPLC) Assay

[0207] A HPLC assay method was developed and optimized to determine specificity, linearity, accuracy, and repeatability of FSD solutions with concentrations ranging from 10 to lOO mg / L.

[0208] Chromatographic parameters were defined using a standard solution of FSD at 100 mg / L. Chromatogram and characteristic values of the FSD peak at 216 nm are shown in FIG.3. The 2D and 3D UV spectra of the FSD peak are shown in FIGs. 4A-4B. The spectra were recorded from 200 to 500 nm. FSD has maximum absorbance at 216 nm in the mobile phase. Specificity was verified by comparing the chromatogram of a FSD standard solution (FIG. 5A) to a chromatogram of a FSD standard solution in the presence of excipients (FIG. 5B). No interfering peaks coming from excipients were observed.

[0209] A linearity study was carried out with standard solutions and reconstituted standard solutions (formulated FSD) from 10 to 100 mg / L. Each solution was prepared by successive dilution from a single stock solution. Regression lines were calculated, excluding the origin (FIGs. 6A-6B). The response was linear from 10 to 100 mg / L.

[0210] Accuracy was demonstrated for standard and reconstituted standard solutions. The recovery of each individual sample was determined by comparing the values of the reconstituted standard solutions with the linear regression curve of the standard solutions.Results are found in Table 4. Individual recoveries varied between 98.18% and 100.94% and the mean recovery (99.71%) was within 98%-102%.Table 4: Accuracy

[0211] Repeatability of responses was assessed by six injections of 100 mg / L reconstituted standard solution. The relative standard deviation of the FSD peak areas was calculated. Results are found in Table 5. The %RSD was less than 2.0%.Table 5: Repeatability

[0212] The HPLC method was qualified for the analysis of FSD solutions. The HPLC method demonstrated specificity and repeatability. Linearity and accuracy were confirmed with FSD solutions from 10 mg / L to 100 mg / L in both the absence and the presence of excipients.

[0213] In vivo characterization of fasudil

[0214] FSD hydrochloride was administered to rats with myocardial ischemia and reperfusion at 10 mg / kg; intravenous injection; 1 hour before operation. Results showed that FSD hydrochloride exhibited protectable effects on cardiovascular disease and reduced the activation of JNK and attenuated mitochondrial-nuclear translocation of AIF under ischemic injury.

[0215] FSD hydrochloride (50 mg / kg / d; i.p.) inhibited acute and relapsing EAE (experimental autoimmune encephalomyelitis) induced by proteolipid protein PLP pl39-151, reduced lymphocyte proliferation, and resulted in down regulation of interleukin (IL)-l 7 and a marked decrease of the IFN-y / IL-4 ratio.

[0216] FSD hydrochloride (100 mg / kg / d; p.o.) significantly reduced incidence and pathological examination score of EAE (experimental autoimmune encephalomyelitis) in SJL / J mice, and decreased inflammation, demyelination, axonal loss and APP positive in spinal cord in mice.EXAMPLE 2: Sublingual Tablets of Fasudil

[0217] Preparation of Sublingual Tablets

[0218] FSD hydrochloride, cellulose, microcrystalline, mannitol, and sodium starch glycolate were accurately massed according to the proportions shown in Table 6.Table 6: FSD Sublingual Tablet

[0219] The powders were combined and mixed homogeneously by trituration in a glass mortar with a pestle using geometric dilution. Then magnesium stearate was added for lubrication and triturated well. The blended material was compressed into 150 mg tablets having a thickness of 8 mm.

[0220] Placebo Preparation

[0221] Cellulose microcrystafline, mannitol, and sodium starch glycolate were accurately massed according to the proportions shown in Table 7.Table 7: Placebo Sublingual Tablet

[0222] Powders were combined and mixed homogeneously by trituration in a glass mortar with a pestle using geometric dilution. Then magnesium stearate was added for lubrication and triturated well. The blended material was compressed into 150 mg tablets having thickness of 8 mm.

[0223] Evaluation of Sublingual Tablets

[0224] Ten randomly chosen FSD and placebo tablets were taken, and the thickness and diameter were determined with a vernier caliper.

[0225] Compatibility between FSD and excipients was carried out by Differential Thermal Analysis (DTA). Approximately 8-13 mg of each sample of FSD, placebo and formulation were placed in a platinum pan of 100 pL capacity. Samples were heated from 25 °C to 600 °C at a rate of 10 °C7min-1under air flow. Indium, aluminum, and zinc were used as standards for temperature calibration.

[0226] 20 tablets from each formulation were randomly taken and accurately massed to test tablet mass variation. The average mass of the tablets was determined. The individual tablet masses were compared with average mass.

[0227] Ten randomly chosen FSD tablets were assayed individually to test uniformity of drug content. Each tablet was powdered and suspended in water in a 50 mL volumetric flask. The active ingredient was extracted by a continuous stirring. The contents of the flask were spun by centrifuge at 8000 G for 10 minutes and fdtered through 0.45 pm PVDF membrane. After suitable dilution, the solution was assayed by HPLC according to the developed method described previously. Drug content was expressed as a percentage of claimed labels and should be 100±15%.

[0228] A shake test was performed to evaluate the disintegration time of FSD and placebo tablets. A tablet was placed in a 20 mL glass beaker containing 2 mL of water. The beaker was manually shaken back and forth to disperse the tablet. The time needed for the tablet disintegration into fine particles was recorded. Tests were performed in triplicate.

[0229] In vitro dissolution tests were performed according to a method specifically designed for rapidly disintegrating sublingual tablet dosage forms. This method simulates the sublingual cavity conversely to the official compendium method, which uses large volumes of dissolution medium with constant agitation around the tablets. A schematic diagram of the apparatus and a brief description of the in vitro dissolution method are shown in FIG. 7.

[0230] A 0.45 pm PVDF membrane was pre-wetted with distilled water and placed between the 15 mL glass funnel and the fritted glass base, which were clamped and inserted into a Buchner flask. A 10 mL disposable plastic collection tube was placed at the outlet tip of the clamped unit to collect the filtrate. The Buchner flask was connected to a vacuum line controlled by automatic shut-off, quick-disconnect coupling inserts (on / off switches).

[0231] 2 mL of distilled water at 25 °C, as a dissolution medium, was added into the 15 mL glass funnel. The tablet was dipped in the dissolution medium undisturbed. Time points from30 to 120 seconds, were previously tested to assess the dissolution rate of a representative formulation of FSD tablets. Based on these results, the 60 second time-point was selected for the subsequent experiments. At each time-point, the full vacuum was applied by opening the on / off switch causing the total volume of dissolution medium to be withdrawn instantly through a 0.45 pm membrane into the collection tube and terminating any further dissolution. The membrane prevented the passage of any undissolved particles and was replaced by a new membrane for each dissolution analysis. Then, each sample was analyzed by HPLC. Undissolved particles retained by the membrane were also analyzed by HPLC.

[0232] The reusable parts including the glass funnel and the fritted glass base were thoroughly cleaned between tests. Any FSD remaining in the dissolution apparatus was totally removed prior to the next dissolution test in order to maintain the integrity of testing.

[0233] Results

[0234] The direct compression technique was used for the preparation of fast disintegrating sublingual tablets of FSD. Sodium starch glycolate was used as superdisintegrant, mannitol as binder, cellulose microcrystaHine as filler, and magnesium stearate as lubricant, in the proportions reported in Table 8.Table 8: Composition of the FSD Tablet

[0235] The FSD dose was 40 mg (e.g., 45 mg of FSD hydrochloride) in a 150 mg tablet. FSD and placebo tablets were off white, circular, biconcave, and odorless with the following geometry: Diameter ranging from 8.09 mm to 8.14 mm, and athickness ranging from 3.84 mm to 3.92 mm (FIG. 8).

[0236] FSD, mixture of excipients, and formulated FSD were sampled and analyzed by DTA. Analyses were carried out under air. Temperature was increased from room temperature to 600 °C at 10 °C / minute.

[0237] At 600 °C, the mass losses were 93.87%, 97.2%, and 98.43% for FSD hydrochloride, excipients, and formulated FSD respectively. FIG. 9 shows that the mass loss profde after 200 °C was similar for the two ampules containing FSD. The mass losses before 150 °C were due to residual water evaporation.

[0238] Thermograms recorded during DTA measurements are shown in FIG. 10. FSD exhibited two endothermic peaks at 138.08 °C referring to water evaporation and at 220.66 °C referring to the FSD melting.

[0239] Endothermic peaks observed at 158.20 °C and 168.80 °C on “Formulated FSD” and “excipients” , respectively, correspond to the melting of mannitol. On the “Formulated FSD” curve, the endothermic peak that corresponds to the melting of FSD (220.66°C) did not occur. This observation was not necessarily characteristic of an incompatibility between FSD and excipients as mannitol melts before FSD and dissolution of FSD it is possible.

[0240] For tablets massing between 80-250 mg, a deviation of less than 7.5% was sought.Twenty tablets were massed individually. Results are shown in FIG. 11. Prepared tablets had uniform mass ranging from 148.69 mg to 151.02 mg with a %RSD = 0.52%.

[0241] Drug content uniformity was tested on ten tablets (FIG. 12). Drug Content is expressed in mg of FSD per tablet and in percentage of the claimed label (40 mg). %RSD was found to be 97.5 ± 3.3 % and proved satisfactory and uniform distribution of FSD in all tablets.

[0242] Shake tests were performed as previously described. Results are shown in Table 9. Tablets disintegrated into fine particles in about 30 seconds.Table 9: Disintegration time of FSD tablets

[0243] Dissolution in vitro

[0244] The in vitro method used was specifically designed to evaluate the dissolution of rapidly disintegrating sublingual tablets. The method has the advantage of testing dissolution in media volumes as small as 2 mL, which corresponds to a volume of saliva secreted in 2 minutes.

[0245] Dissolution tests were conducted following the methods described herein. The time for retaining sublingual tablets under the tongue is 120 seconds. However, the time-point for collecting the dissolved FSD was set at 60 seconds. The FSD content in the filtrate was compared to the average FSD content in ten tablets (FIG. 13). Almost 90% of FSD was released during dissolution tests within 60 seconds.

[0246] In conclusion, fast dissolving sublingual tablets of FSD were prepared. The direct compression method was used leading to tablets free from chipping, laminating, and capping. DTA tests did not reveal any incompatibility between FSD and the chosen excipients. Mass variation of the produced tablets was lower than 1%. In vitro disintegration time of FSD tablet was found to be about 30 seconds. FSD content in the produced tablets was 97.5% stated on the label (40 mg). Almost 90% of the FSD was released during the dissolution tests.EXAMPLE 3: Bioavailability

[0247] Drugs administered via the sublingual route are directly absorbed into blood circulation, avoiding the hepatic first-pass metabolism. FSD transport through the oral mucosal epithelial barrier was evaluated in vitro using cell culture inserts (e.g., Transwell® cell culture inserts) (FIG. 14). Bioavailability of FSD was studied on SCC4 cell monolayers as an alternative sublingual barrier model. SCC4 cell lines are cells of oral epithelium extracted from human tongue squamous cell carcinoma. Cells were prepared according to the preparation method shown in FIG. 15.

[0248] A stock solution of FSD in HBSS buffer was prepared (FIG. 16A). 45 mg of FSD hydrochloride was massed in a 10 mL volumetric flask. 8 mL of HBSS buffer solution were added, and the suspension was shaken until complete dissolution of the FSD occurred. The volume was completed to the gauge with HBSS. The solution was sterilized by filtration on 0.2 mm PVDF membrane under a BSC laminar flow hood.

[0249] A stock solution of a FSD tablet in HBSS was prepared (FIG. 16B). 8 mL of HBSS buffer solution and one 40 mg FSD tablet were added to a 10 mL volumetric flask. The flask was shaken and sonicated until a homogeneous suspension was obtained. The volume was completed to the gauge with HBSS. The solution was filtered on 0.2 mm PVDF membrane under a MSC laminar flow hood.

[0250] A stock solution of the placebo was prepared (FIG. 16C). 8 mL of HBSS buffer solution and 1 Placebo FSD tablet were added to a 10 mL volumetric flask. The flask was shaken and sonicated until a homogeneous suspension was obtained. The volume was completed to the gauge with HBSS. The solution was filtered on 0.2 mm PVDF membrane under a MSC laminar flow hood.

[0251] Fasudil tablet and placebo tablet suspensions were either filtered or not filtered before filling the inserts (FIG. 17A-17C). The culture media was removed from each insert and well. The apical compartment was rinsed with 500 pL HBSS lx. The basolateral compartment was rinsed with 500 pL HBSS lx. The two compartments were emptied and 500 pL HBSS lx was added to both the apical compartment and the basolateral compartment. Cell monolater resistance was measured (R at t=0), and the two compartments were emptied. 500 pL HBSS lx was added to each basolateral compartment and 500 pL of a sample was added to a well. The concentration of FSD was followed by HPLC in each compartment at t = 0, 15, 30, 60, and 180 minutes. After 180 minutes, the solutions of each compartment were individually collected. 500 pL HBSS lx was added to both the apical compartment and the basolateral compartment and the resistance of the cell monolayers was measured (R at t=0).

[0252] Transepithelial / transendothelial electrical resistance (TEER) is accepted as a quantitative technique to measure the integrity of tight junction dynamics in cell culture models of epithelial monolayers. Cell monolayer resistances were measured before and after the bioavailability tests. Results are shown in FIGs. 18A-18B.

[0253] R determined the resistance to ion flux between the apical and basolateral sides of the cell monolayer and therefore the integrity of the intercellular tight junctions. TEER values represented the resistance per surface unit corrected by subtraction of the background (“Control without cell monolayer”). Obtained TEER values were close to those reported for other oral epithelial cells (H376 TEER = 31.40 ±4.28 Q cm’2; TR146 TEER = 50.02 ±2.87 Q cm’2). Under experimental conditions, the integrity of the cell monolayers was maintained all along the tests.

[0254] Analytical results of both apical and basolateral compartments during the transepithelial transfer of FSD are shown in FIGs. 19A-19B. Apparent permeability Papp was used to determine the degree of permeability to rank different drug substances and to comprehend transportation pathways. The Papp is calculated using the equation:

[0255] where dQ / dt is the slope of the cumulative fraction absorbed versus time in seconds, A is the area of the filter in cm2, and Co is the initial concentration in the apical chamber. Drug permeability according to Papp is summarized in Table 10. Calculated Papp varied between 7.6*1 O'6to 9*1 O'6cm / s, indicating that FSD is part of the standard permeability drugs.Table 10: Apparent drug permeability index

[0256] The kinetics of the transepithelial transfer of FSD through SCC-4 monolayer is shown in FIGs. 20A-20C. The kinetic transfer was linear. From 15 to 180 minutes, the transfer rate from the apical chamber to the basolateral chamber was V = 0.09 pg / min*cm2. At the initial times, no difference was observed between the filtered and the non-filtered suspensions.When the initial concentration of FSD in the apical chamber was [FSD]=1 000 mg / L, the transfer rate was V = 0.49 pg / min*cm2.

[0257] In conclusion, SCC-4 cell line (oral epithelium extracted from human tongue squamous cell carcinoma) were suitable for modeling the sublingual bioavailability of FSD. Excipients had no deleterious effect on the cell monolayer. The transfer kinetics of FSD corresponded to what is expected of a standard transfer (0.5 pg / minute). No difference was observed between the bioavailability of FSD alone and the FSD tablet.EXAMPLE 4: Therapy

[0258] A subject is diagnosed with ALS. The subject is prescribed a sublingual tablet as described herein for ALS therapy. The subject begins a course of therapy for ALS by taking a unit dosage form sublingually on a prescribed schedule.

Claims

CLAIMSWHAT IS CLAIMED IS:

1. A pharmaceutical composition comprising fasudil or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition is a sublingual dosage form.

2. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition further comprises a pharmaceutically-acceptable excipient.

3. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition further comprises cellulose.

4. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition further comprises mannitol.

5. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition further comprises sodium starch glycolate.

6. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition further comprises magnesium stearate.

7. The pharmaceutical composition of claim 1, wherein the fasudil or the pharmaceutically-acceptable salt thereof is present in the pharmaceutical composition in a concentration of about 50 nM to about 300 nM.

8. The pharmaceutical composition of claim 1, wherein the fasudil or the pharmaceutically acceptable salt thereof is present in the pharmaceutical composition in a concentration of at least 1 pM.

9. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition comprises the pharmaceutically-acceptable salt, and the pharmaceutically acceptable salt is a hydrochloride salt.

10. The pharmaceutical composition of claim 1, wherein the fasudil or the pharmaceutically acceptable salt thereof is present in the pharmaceutical composition in an amount of about 20 mg to about 60 mg.

11. The pharmaceutical composition of claim 1, wherein the fasudil or the pharmaceutically acceptable salt thereof is present in the pharmaceutical composition in an amount of about 40 mg to about 50 mg.

12. The pharmaceutical composition of claim 1, wherein the fasudil or the pharmaceutically acceptable salt thereof is present in the pharmaceutical composition in an amount of about 45 mg.

13. The pharmaceutical composition of claim 1, further comprising a filler, a binder, asuperdisintegrant, and a lubricant.

14. The pharmaceutical composition of claim 1, further comprising microcrystalline cellulose, mannitol, sodium starch glycolate, and magnesium stearate.

15. The pharmaceutical composition of claim 1, further comprising: a) microcrystalline cellulose in an amount of about 20 mg to about 40 mg; b) mannitol in an amount of a about 20 mg to about 100 mg; c) sodium starch glycolate in an amount of about 1 mg to about 20 mg; and d) magnesium stearate in an amount of about 0.5 mg to about 10 mg.

16. The pharmaceutical composition of claim 1, further comprising: a) microcrystalline cellulose in an amount of about 33 mg; b) mannitol in an amount of a about 60 mg; c) sodium starch glycolate in an amount of about 9 mg; and d) magnesium stearate in an amount of about 3 mg.

17. The pharmaceutical composition of claim 1, wherein the sublingual dosage form is a tablet.

18. The pharmaceutical composition of claim 1, wherein the sublingual dosage form is a tablet that is round and biconcave.

19. The pharmaceutical composition of claim 1, wherein the sublingual dosage form is a tablet that has a diameter from about 7 mm to about 9 mm and a thickness from about 3.5 mm to about 4.5 mm.

20. The pharmaceutical composition of claim 1, wherein the sublingual dosage form is a tablet that has a diameter from about 8 mm to about 8.2 mm and a thickness from about 3.8 mm to about 4 mm.

21. The pharmaceutical composition of claim 1, wherein the sublingual dosage form is solid, wherein if the sublingual dosage form is subjected to a shake test in which the sublingual dosage form is placed in a glass beaker containing 2 mL of water and the beaker is manually shaken to disintegrate the sublingual dosage form in the water, then the time needed for disintegration of the sublingual dosage form into fine particles is less than sixty seconds.

22. The pharmaceutical composition of claim 1, wherein the sublingual dosage form is solid, wherein if the sublingual dosage form is subjected to a shake test in which the sublingual dosage form is placed in a glass beaker containing 2 mL of water and the beaker is manually shaken to disintegrate the sublingual dosage form in the water, then the time needed for disintegration of the sublingual dosage form into fine particles is less than forty seconds.

23. The pharmaceutical composition of claim 1, wherein the sublingual dosage form issolid, wherein if the sublingual dosage form is subjected to a shake test in which the sublingual dosage form is placed in a glass beaker containing 2 mL of water and the beaker is manually shaken to disintegrate the sublingual dosage form in the water, then the time needed for disintegration of the sublingual dosage form into fine particles is 30 ± 2 seconds.

24. The pharmaceutical composition of claim 1, wherein if a sample of about 8 mg to about 13 mg of the sublingual dosage form is subjected to differential thermal analysis by placing the sample in a platinum pan of 100 pL capacity, and the sample is heated from 25 °C to 600 °C at a rate of 10 °C / min'1under air flow, then a resulting thermograph shows an endothermic peak at 145-170 °C and does not show an endothermic peak at 210-235 °C.

25. The pharmaceutical composition of claim 1, wherein if a sample of about 8 mg to about 13 mg of the sublingual dosage form is subjected to differential thermal analysis by placing the sample in a platinum pan of 100 pL capacity, and the sample is heated from 25 °C to 600 °C at a rate of 10 °C / min'1under air flow, then a resulting thermograph shows an endothermic peak at 156-161 °C and does not show an endothermic peak at 218-223 °C.

26. The pharmaceutical composition of claim 1, wherein the sublingual dosage form is solid, wherein if the sublingual dosage form is subjected to a dissolution study in which the sublingual dosage form is dipped and held in a medium of 2 mL of distilled water at 25 °C in a 15 mL glass funnel for sixty seconds, at which point the medium is drawn by vacuum through a 0.45 pm membrane into a collection tube, after which the medium is analyzed by HPLC, then a percent dissolution of fasudil or the pharmaceutically acceptable salt thereof of 80% - 100% is obtained.

27. The pharmaceutical composition of claim 1, wherein the sublingual dosage form is solid, wherein if the sublingual dosage form is subjected to a dissolution study in which the sublingual dosage form is dipped and held in a medium of 2 mL of distilled water at 25 °C in a 15 mL glass funnel for sixty seconds, at which point the medium is drawn by vacuum through a 0.45 pm membrane into a collection tube, after which the medium is analyzed by HPLC, then a percent dissolution of fasudil or the pharmaceutically acceptable salt thereof of 85% - 95% is obtained.

28. The pharmaceutical composition of claim 1, wherein the sublingual dosage form is solid, wherein if the sublingual dosage form is subjected to a dissolution study in which the sublingual dosage form is dipped and held in a medium of 2 mL of distilled water at 25 °C in a 15 mL glass funnel for sixty seconds, at which point the medium is drawn by vacuum through a 0.45 pm membrane into a collection tube, after which the medium is analyzed by HPLC, then a percent dissolution of fasudil or the pharmaceutically acceptable salt thereof of89.6 ± 4.0% is obtained.

29. A pharmaceutical composition comprising fasudil or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition is a solid dosage form, wherein if the solid dosage form is subjected to a shake test in which the solid dosage form is placed in a glass beaker containing 2 mL of water and the beaker is manually shaken to disintegrate the solid dosage form in the water, then the time needed for disintegration of the solid dosage form into fine particles is less than sixty seconds.

30. The pharmaceutical composition of claim 29, wherein the time needed for disintegration of the solid dosage form into fine particles is less than forty seconds.

31. The pharmaceutical composition of claim 29, wherein the time needed for disintegration of the solid dosage form into fine particles is 30 ± 2 seconds.

32. The pharmaceutical composition of claim 29, wherein the pharmaceutical composition further comprises a pharmaceutically-acceptable excipient.

33. The pharmaceutical composition of claim 29, wherein the pharmaceutical composition further comprises cellulose.

34. The pharmaceutical composition of claim 29, wherein the pharmaceutical composition further comprises mannitol.

35. The pharmaceutical composition of claim 29, wherein the pharmaceutical composition further comprises sodium starch glycolate.

36. The pharmaceutical composition of claim 29, wherein the pharmaceutical composition further comprises magnesium stearate.

37. The pharmaceutical composition of claim 29, wherein the fasudil or the pharmaceutically-acceptable salt thereof is present in the pharmaceutical composition in a concentration of about 50 nM to about 300 nM.

38. The pharmaceutical composition of claim 29, wherein the fasudil or the pharmaceutically acceptable salt thereof is present in the pharmaceutical composition in a concentration of at least 1 pM.

39. The pharmaceutical composition of claim 29, wherein the pharmaceutical composition comprises the pharmaceutically-acceptable salt, and the pharmaceutically acceptable salt is a hydrochloride salt.

40. The pharmaceutical composition of claim 29, wherein the fasudil or the pharmaceutically acceptable salt thereof is present in the pharmaceutical composition in an amount of about 20 mg to about 60 mg.

41. The pharmaceutical composition of claim 29, wherein the fasudil or thepharmaceutically acceptable salt thereof is present in the pharmaceutical composition in an amount of about 40 mg to about 50 mg.

42. The pharmaceutical composition of claim 29, wherein the fasudil or the pharmaceutically acceptable salt thereof is present in the pharmaceutical composition in an amount of about 45 mg.

43. The pharmaceutical composition of claim 29, further comprising a filler, a binder, a superdisintegrant, and a lubricant.

44. The pharmaceutical composition of claim 29, further comprising microcrystaHine cellulose, mannitol, sodium starch glycolate, and magnesium stearate.

45. The pharmaceutical composition of claim 29, further comprising: a) microcrystalline cellulose in an amount of about 20 mg to about 40 mg; b) mannitol in an amount of a about 20 mg to about 100 mg; c) sodium starch glycolate in an amount of about 1 mg to about 20 mg; and d) magnesium stearate in an amount of about 0.5 mg to about 10 mg.

46. The pharmaceutical composition of claim 29, further comprising: a) microcrystalline cellulose in an amount of about 33 mg; b) mannitol in an amount of a about 60 mg; c) sodium starch glycolate in an amount of about 9 mg; and d) magnesium stearate in an amount of about 3 mg.

47. The pharmaceutical composition of claim 29, wherein the solid dosage form is a tablet.

48. The pharmaceutical composition of claim 29, wherein the solid dosage form is a tablet that is round and biconcave.

49. The pharmaceutical composition of claim 29, wherein the solid dosage form is a tablet that has a diameter from about 7 mm to about 9 mm and a thickness from about 3.5 mm to about 4.5 mm.

50. The pharmaceutical composition of claim 29, wherein the solid dosage form is a tablet that has a diameter from about 8 mm to about 8.2 mm and a thickness from about 3.8 mm to about 4 mm.

51. A pharmaceutical composition comprising fasudil or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition is a solid dosage form, wherein if a sample of about 8 mg to about 13 mg of the solid dosage form is subjected to differential thermal analysis by placing the sample in a platinum pan of 100 pL capacity, and the sample is heated from 25 °C to 600 °C at a rate of 10 °C / min'1under air flow, then a resultingthermograph shows an endothermic peak at 145-170 °C and does not show an endothermic peak at 210-235 °C.

52. The pharmaceutical composition of claim 51, wherein the resulting thermograph shows an endothermic peak at 156-161 °C and does not show an endothermic peak at 218- 223 °C.

53. The pharmaceutical composition of claim 51, wherein the pharmaceutical composition further comprises a pharmaceutically-acceptable excipient.

54. The pharmaceutical composition of claim 51, wherein the pharmaceutical composition further comprises cellulose.

55. The pharmaceutical composition of claim 51, wherein the pharmaceutical composition further comprises mannitol.

56. The pharmaceutical composition of claim 51, wherein the pharmaceutical composition further comprises sodium starch glycolate.

57. The pharmaceutical composition of claim 51, wherein the pharmaceutical composition further comprises magnesium stearate.

58. The pharmaceutical composition of claim 51, wherein the fasudil or the pharmaceutically-acceptable salt thereof is present in the pharmaceutical composition in a concentration of about 50 nM to about 300 nM.

59. The pharmaceutical composition of claim 51, wherein the fasudil or the pharmaceutically acceptable salt thereof is present in the pharmaceutical composition in a concentration of at least 1 pM.

60. The pharmaceutical composition of claim 51, wherein the pharmaceutical composition comprises the pharmaceutically-acceptable salt, and the pharmaceutically acceptable salt is a hydrochloride salt.

61. The pharmaceutical composition of claim 51, wherein the fasudil or the pharmaceutically acceptable salt thereof is present in the pharmaceutical composition in an amount of about 20 mg to about 60 mg.

62. The pharmaceutical composition of claim 51, wherein the fasudil or the pharmaceutically acceptable salt thereof is present in the pharmaceutical composition in an amount of about 40 mg to about 50 mg.

63. The pharmaceutical composition of claim 51, wherein the fasudil or the pharmaceutically acceptable salt thereof is present in the pharmaceutical composition in an amount of about 45 mg.

64. The pharmaceutical composition of claim 51, further comprising a fdler, a binder, asuperdisintegrant, and a lubricant.

65. The pharmaceutical composition of claim 51, further comprising microcrystalline cellulose, mannitol, sodium starch glycolate, and magnesium stearate.

66. The pharmaceutical composition of claim 51, further comprising: a) microcrystalline cellulose in an amount of about 20 mg to about 40 mg; b) mannitol in an amount of a about 20 mg to about 100 mg; c) sodium starch glycolate in an amount of about 1 mg to about 20 mg; and d) magnesium stearate in an amount of about 0.5 mg to about 10 mg.

67. The pharmaceutical composition of claim 51, further comprising: a) microcrystalline cellulose in an amount of about 33 mg; b) mannitol in an amount of a about 60 mg; c) sodium starch glycolate in an amount of about 9 mg; and d) magnesium stearate in an amount of about 3 mg.

68. The pharmaceutical composition of claim 51, wherein the solid dosage form is a tablet.

69. The pharmaceutical composition of claim 51, wherein the solid dosage form is a tablet that is round and biconcave.

70. The pharmaceutical composition of claim 51, wherein the solid dosage form is a tablet that has a diameter from about 7 mm to about 9 mm and a thickness from about 3.5 mm to about 4.5 mm.

71. The pharmaceutical composition of claim 51, wherein the solid dosage form is a tablet that has a diameter from about 8 mm to about 8.2 mm and a thickness from about 3.8 mm to about 4 mm.

72. A method for treating a condition, the method comprising sublingually administering to a subject in need thereof a therapeutically-effective amount of fasudil or a pharmaceutically acceptable salt thereof.

73. A method for treating a condition, the method comprising administering to a subject in need thereof a therapeutically-effective amount of the pharmaceutical composition of any one of claims 1-71.

74. The method of any one of claims 72 or 73, wherein the condition is a neurodegenerative disease.

75. The method of any one of claims 72-74, wherein the condition is a central nervous system disease.

76. The method of any one of claims 72-75, wherein the condition is an aging condition.

77. The method of any one of claims 72-76, wherein the condition is amyotrophic lateral sclerosis (ALS).

78. The method of any one of claims 72-76, wherein the condition is dementia.

79. The method of any one of claims 72-76, wherein the condition is Alzheimer’sDisease.

80. The method of any one of claims 72-76, wherein the condition is Parkinson’s Disease.

81. The method of any one of claims 72-80, wherein the therapeutically-effective amount is about 20 mg to about 60 mg.

82. The method of any one of claims 72-81, wherein the therapeutically-effective amount is about 40 mg to about 50 mg.

83. The method of any one of claims 72-82, wherein the therapeutically-effective amount is about 45 mg.

84. The method of any one of claims 72-83, wherein treating the condition reduces muscle weakness in the subject by at least 10%.

85. The method of any one of claims 72-83, wherein treating the condition reduces motor neuron loss in the subject by at least 10%.

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