Leucines, acetylleucines and related analogs for treatment of disease

Abstract

The present disclosure relates to leucines, acetyl leucines and related analogs for the treatment of disease. The present disclosure provides methods of treating a disease, disorder, or condition in a patient in need thereof by administering a therapeutically effective amount of acetyl-L-leucine, or a pharmaceutically acceptable salt thereof.

Description

[0001] This application is a divisional application. The international application number of the original application is PCT / IB2020 / 051767, the application date is March 2, 2020, the application number of the Chinese national phase is 202080029158.5, and the name of the invention is "Leucine, acetylleucine and related analogs for treating diseases". Technical Field

[0002] The present disclosure provides methods of treating a disease, condition, disorder or syndrome, such as traumatic brain injury, in a patient in need thereof by administering a therapeutically effective amount of DL-leucine or a pharmaceutically acceptable salt thereof, D-leucine or a pharmaceutically acceptable salt thereof, L-leucine or a pharmaceutically acceptable salt thereof, L-leucine ethyl ester or a pharmaceutically acceptable salt thereof, acetyl-DL-leucine or a pharmaceutically acceptable salt thereof, acetyl-D-leucine or a pharmaceutically acceptable salt thereof, or acetyl-L-leucine or a pharmaceutically acceptable salt thereof. Background Art

[0003] Acetyl-DL-leucine has been used in France to treat acute vertigo since 1957. An FDG-μPET study in an acute unilateral labyrinthectomy rat model (Zwergal et al. (2016) Brain Struct Funct; 221(1): 159-70) showed a significant effect of the L-enantiomer, N-acetyl-L-leucine, on postural compensation by activating the vestibulocerebellum and inactivating the posterolateral thalamus (Gunther et al. (2015) PLoS One; 10(3): e0120891). In a case series of cerebellar patients, symptomatic improvement of cerebellar ataxia with the use of acetyl-DL-leucine was shown (Strupp et al. (2013) J Neurol; 260(10): 2556-61). Another case series found no benefit (Pelz et al. (2015) J Neurol; 262(5): 1373-5). Quantitative gait analysis showed that acetyl-DL-leucine improved temporal gait variability in patients with cerebellar ataxia (Schniepp et al. (2015) Cerebellum; 3:8). In a one-month study involving 12 patients with Niemann-Pick type C (NPC), symptomatic improvement of ataxia was shown (Bremova et al. (2015) Neurology; 85(16): 1368-75). In addition, PET studies in patients with ataxia given acetyl-DL-leucine showed increased metabolism in the midbrain and lower brainstem in responders (Becker-Bense et al. (2015) Abstract EAN).

[0004] WO 2018 / 029657 and WO 2018 / 029658 describe the use of leucine and acetylleucine for the treatment of certain neurodegenerative diseases such as Alzheimer's disease (AD), multiple sclerosis (MS), Parkinson's disease (PD) and amyotrophic lateral sclerosis (ALS) and certain lysosomal storage diseases (LSD). In WO 2018 / 029658, a Niemann-Pick disease type C (NPC) mouse model showed that NPC1 mouse brains exhibited accumulation of LC3-II, and administration of acetyl-DL-leucine was associated with a decrease in LC3-II, indicating that autophagic flux was partially restored.

[0005] Inflammation caused by the production of proinflammatory cytokines (e.g., IL-1β) and chemokines is the cornerstone of various neurodegenerative, neuroinflammatory, mitochondrial and other diseases. See, e.g., Chakrabarti et al., Curr Alzheimer Res. 15(10):894–904 (2018); Cherry et al., Journal of Neuroinflammation 11:98 (2014); Inácio et al., Journal of Neuroinflammation 12:211 (2015); Lappalainen et al., Am J Respir Cell Mol Biol 32:311–318 (2005); Coccia et al., J. Exp. Med. 209:1595-1609 (2012); Ray, Journal of the American College of Cardiology 63:1735-1438 (2014); Ren et al., Brain Res Rev. 60(1):57–64 (2009); Frank-Cannon et al., Molecular Neurodegeneration 4:47 (2009); DiSabato et al., J Neurochem 139(Suppl 2):136–153 (2016); Amor et al., Immunology 142:151–166 (2013); Chen et al., Molecular Medicine Reports 13:3391-3396 (2016); Tuttolomondo et al., Drug Design, Development and Therapy 8:2221–2239 (2014); Hong et al., Int Neurourol J 20 Suppl 1:S2-7 (2016); Gu et al., Journal of Biological Regulators & Homeostatic Agents 29:787-791 (2015); Belarbi et al., Journal of Neuroinflammation 9:23 (2012); Abdulkhaleq et al., Veterinary World, EISSN: 2231-0916 (available at www.veterinaryworld.org / Vol.11 / May-2018 / 9.pdf); Frankola et al., CNS Neurol Disord Drug Targets 10(3):391–403 (2011); Habbas et al., Cell 163:1730–1741 (2015); Debray et al., Current Opinion in Pediatrics 20:471–482 (2008); Niyazov et al., Mol Syndromol 7:122–137 (2016). .

[0006] For example, traumatic brain injury ("TBI") is a devastating condition associated with high morbidity and mortality. Given population growth and the increase in injury-related events such as traffic accidents, the incidence of TBI continues to rise. Other emergency situations, such as natural disasters, sports injuries, falls, assaults, and military conflicts contribute significantly to TBI cases. In the United States alone, it is estimated that more than 1.5 million people suffer TBI each year. The severity of TBI varies, ranging from mild, moderate to severe, and can be focal or diffuse. Focal injuries occur in specific locations, while diffuse injuries are associated with potentially widespread axonal dysfunction and vascular damage.

[0007] TBI damage to the brain occurs in two phases. The initial primary phase is the head trauma event itself, which is irreversible and only preventive measures can be taken to minimize the extent of the damage, followed by a persistent second phase that begins at the time of injury and persists over the following days, weeks, and years. This second phase results in a variety of physiological, cellular, and molecular responses aimed at restoring homeostasis of damaged tissue, which may lead to secondary brain injury.

[0008] There is a need in the art for agents to treat diseases responsive to modulation of the expression of pro-inflammatory mediators. Summary of the invention

[0009] The applicant unexpectedly found that acetyl-L-leucine regulates the expression of proinflammatory mediators in a controlled cortical impact-induced (CCI) traumatic brain injury model in mice. Therefore, in one aspect, the present disclosure provides a method for treating a disease, disorder, condition or syndrome, wherein the regulation of one or more proinflammatory mediators provides benefits. Among these diseases, disorders, conditions or syndromes, the most important are neuroinflammatory, nervous system, autoimmune, neurodegenerative protein diseases, psychosis and mitochondrial diseases, disorders, conditions or syndromes.

[0010] Without wishing to be bound by any particular theory, it is believed that leucine, acetyl-DL-leucine, acetyl-D-leucine, and acetyl-L-leucine modulate neuroinflammation, which is a pathway to counteract the molecular cascades that lead to neuroinflammatory and other diseases, disorders, conditions, or syndromes. For example, secondary brain injury in patients with TBI can be characterized by an overactive response, in which excessive proinflammatory cytokines can play a role in the maintenance of brain function and repair after TBI. Brain injury induced by head trauma can trigger excessive and / or uncontrolled release of these cytokines, particularly interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α, which can lead to severe brain damage. It is reported that proinflammatory cytokines play a role in the pathophysiology of neurodegenerative diseases such as AD, MS, PD and ALS (Wei-Wei et al., Mol. Med. Rep. 2016; 13(4): 3391-3396), and have been identified as a hallmark of hereditary-neurodegenerative disorders such as GM1 and GM2 gangliosides (Jeyakumar et al., Brain 2003; 126(Pt4): 974-87). In addition, disruption of normal lysosomal function in lysosomal storage diseases (LSDs) has been associated with inflammatory abnormalities such as abnormally elevated proinflammatory cytokines. (Simonaro, Journal of Inborn Errors of Metabolism & Screening, Vol. 4: 1-8 (2016)).

[0011] In addition, without wishing to be bound by any particular theory, it is also believed according to the present disclosure that DL-leucine, L-leucine, D-leucine, L-leucine ethyl ester, acetyl-DL-leucine, acetyl-D-leucine and acetyl-L-leucine regulate microtubule-associated protein Tau (which is an autophagy substrate) and / or microtubule-associated protein 1A / 1B-light chain 3-phosphatidylethanolamine conjugate (LC3-II), which can be a marker of TBI. LC3-II is a marker of autophagosome formation, and elevated levels of LC3-II can reflect impaired clearance of autophagic vacuoles. Autophagosomes are formed, but not cleared. Autophagy is impaired in AD, and AD brains show elevated LC3-II levels.

[0012] In one aspect, the disclosure provides a method of treating a disease, condition, disorder or syndrome in a subject in need thereof, or treating a symptom of a disease, condition, disorder or syndrome in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of the disclosure, wherein the disease, condition, disorder or syndrome is any one or more of the diseases listed in Tables 1-17, see below. "Compounds of the disclosure" are collectively referred to as DL-leucine ("DLL"), L-leucine ("LL"), D-leucine ("DL"), L-leucine ethyl ester ("LLE" or "LEE"), acetyl-DL-leucine ("ADLL"), acetyl-D-leucine ("ADL") or acetyl-L-leucine ("ALL", "NAL" or "NALL"), and pharmaceutically acceptable salts thereof.

[0013] In another aspect, the disclosure provides a method of treating TBI in a subject in need thereof or treating a symptom of TBI in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of the disclosure.

[0014] In another aspect, the present disclosure provides a compound of the present disclosure for use in treating a disease, disorder, condition or syndrome in a subject in need thereof, or for treating a symptom of a disease, disorder, condition or syndrome in a subject in need thereof, wherein the disease, disorder, condition or syndrome is any one or more of the diseases listed in Tables 1-17, see below.

[0015] In another aspect, the disclosure provides a compound of the disclosure for use in treating TBI in a subject in need thereof or for treating a symptom of TBI in a subject in need thereof.

[0016] In another aspect, the disclosure provides the use of a compound of the disclosure for the manufacture of a medicament for treating a disease, disorder, condition or syndrome in a subject in need thereof, or for the manufacture of a medicament for treating a symptom of a disease, disorder, condition or syndrome in a subject in need thereof, wherein the disease, disorder, condition or syndrome is any one or more of the diseases listed in Tables 1-17, see below.

[0017] In another aspect, the disclosure provides the use of a compound of the disclosure for the manufacture of a medicament for treating TBI in a subject in need thereof or for the manufacture of a medicament for treating TBI in a subject in need thereof.

[0018] In another aspect, the present disclosure provides a pharmaceutical composition for treating a disease, disorder, condition or syndrome in a subject in need thereof, or for treating a symptom of a disease, disorder, condition or syndrome in a subject in need thereof, the pharmaceutical composition comprising a compound of the present disclosure and a pharmaceutically acceptable carrier, wherein the disease, disorder, condition or syndrome is any one or more of the diseases listed in Tables 1-17, see below.

[0019] In another aspect, the present disclosure provides a method for regulating the expression of proinflammatory mediators (e.g., proinflammatory cytokines and proinflammatory chemokines) in a subject in need thereof, the method comprising administering a therapeutically effective amount of a compound of the present disclosure to the subject. In another aspect, proinflammatory mediators include but are not limited to NOS2, IL-18, IFNb, IL-1β, TNFα, NOX2, NLRP3, SOCS3, ARG1, IL-10, IL-4ra and / or YM1.

[0020] It should be understood that both the foregoing summary and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 is a schematic diagram showing NAL administration and experimental plan for the cortical impact-induced TBI model. Mice were orally fed NAL, and cortical tissues were collected on days 1, 3, and 7 after TBI for biochemical analysis.

[0022] Figure 2 is a bar graph showing the body weight changes of mice 7 days after NAL (N-acetyl-L-leucine or acetyl-L-leucine) treatment after TBI.

[0023] Figure 3 is a bar graph showing the spatial memory assessment of mice after NAL treatment 7 days after TBI using the Y-maze test.

[0024] Figure 4 is a bar graph showing IL-16 expression in mice 3 days after NAL treatment following CCI.

[0025] Figure 5 is a bar graph showing IL-1β expression in mice 3 days after NAL treatment following CCI.

[0026] Figure 6 is a bar graph showing INFb expression in mice 3 days after NAL treatment following CCI.

[0027] Figure 7 is a bar graph showing Arg1 expression in mice 3 days after NAL treatment following CCI.

[0028] Figure 8 is a bar graph showing SOCS3 expression in mice 3 days after NAL treatment following CCI.

[0029] Fig. 9 is a bar graph showing TNF expression in mice 3 days after NAL treatment following CCI.

[0030] Fig.10 is a bar graph showing IL-18 expression in mice 3 days after NAL treatment following CCI.

[0031] Fig.11 Western blot analysis was performed to evaluate autophagy and cell death after NAL treatment following CCI.

[0032] Fig.12 is a bar graph showing fodrin / b-actin levels after NAL treatment following CCI.

[0033] Fig.13 is a bar graph showing LC3-II / b-actin levels after NAL treatment following CCI.

[0034] Fig.14 is a histogram of the ratio of LC3-II expression to βIII-tubulin in the cerebellum of NPC1 brains from 9-12 weeks old mice treated for 3 weeks with ADLL, ALL, ADL, DLL, LL, DL, LLE, and miglu. relative to untreated NPC1 cerebellum (n=4, mean±SEM).

[0035] Fig.15 is a histogram of the ratio of APP-CTF:full-length APP expression in the cerebellum of NPC1 brains from 9-12 week old mice treated for 3 weeks with ADLL, ALL, ADL, DLL, LL, DL, LLE, and miglu. relative to untreated NPC1 cerebellum (n=4, mean±SEM).

[0036] Fig.16 is a histogram of the ratio of APP-CTF-6:full-length APP expression in the cerebellum of NPC1 brains from 9-12 weeks old mice treated for 3 weeks with ADLL, ALL, ADL, DLL, LL, DL, and LLE relative to untreated NPC1 cerebellum.

[0037] Fig.17is a histogram of the ratio of APP-CTF-7:full-length APP expression in the cerebellum of NPC1 brains from 9-12 weeks old mice treated for 3 weeks with ADLL, ALL, ADL, DLL, LL, DL, and LLE relative to untreated NPC1 cerebellum (n=4, mean±SEM).

[0038] Fig.18 is a histogram of the ratio of LC3-II normalized to βIII tubulin in rat primary cortical neurons treated with U18666A (2 μg / ml, 24 hours) in the absence or presence of ADLL, ALL, ADL, DLL, LL, DL and LLE (1 mM, 24 hours), expressed as a percentage of untreated (n=1).

[0039] Fig.19 Histogram of the ratio of APP-CTF-6 and -7 normalized to their respective CTFs in untreated cells in rat primary cortical neurons treated with U18666A (2 μg / ml, 24 h) in the absence or presence of ADLL, ALL, ADL, DLL, LL, DL and LLE (1 mM, 24 h), expressed as a percentage of untreated (n=1).

[0040] Fig. 20 Are eight Western blots of tau (5E2), synaptophysin, and βIII-tubulin in rat primary cortical neurons (neuron-enriched (NE) and neuron-glia (NG) cultures) in the absence and presence of ADLL, ALL, ADL, DLL, LL, DL, and LLE (1 mM, 5 days).

[0041] Fig.21 is a histogram of the ratio of full-length tau normalized to βIII tubulin in NE cultures at DIV14 in the absence and presence of ADLL, ALL, ADL, DLL, LL, DL, and LLE (1 mM, 5 days).

[0042] Fig. 22 is a histogram of the ratio of truncated tau (clone 5E2) normalized to βIII tubulin in NG cultures at DIV14 in the absence and presence of ADLL, ALL, ADL, DLL, LL, DL and LLE (1 mM, 5 days).

[0043] Fig.23is a histogram of the ratio of synaptophysin normalized to βIII tubulin in NE cultures at DIV14 in the absence and presence of ADLL, ALL, ADL, DLL, LL, DL, and LLE (1 mM, 5 days) (expressed as percentage of untreated control lysate; n=4, mean±SEM).

[0044] Fig.24 is a histogram of the ratio of synaptophysin normalized to βIII tubulin in NG cultures at DIV14 in the absence and presence of ADLL, ALL, ADL, DLL, LL, DL, and LLE (1 mM, 5 days) (expressed as percentage of untreated control lysate; n=4, mean±SEM).

[0045] Fig.25 is a bar graph showing lysosomal volume in wild-type and NPC1 -null Chinese hamster ovary cells treated with ALL at the indicated concentrations for 7 days (n=4).

[0046] Fig.26 is a bar graph showing lysosomal volume in wild-type and NPC1 -null Chinese hamster ovary cells treated with LEE at the indicated concentrations for 7 days (n=4).

[0047] Fig. 27 are bar graphs showing lysosomal volumes in wild-type and NPC1 -null Chinese hamster ovary cells treated with LL at the indicated concentrations for 7 days (n=3).

[0048] Fig.28 Western blot analysis of cortical tissue lysates from sham-operated and TBI mice fed NALL or vehicle to detect α-fodrin breakdown products was evaluated.

[0049] Fig.29 is a bar graph showing densitometric analysis of α-fodrin relative to actin. Data are presented as mean±SEM. n=5, **p<0.01 (two-way ANOVA with Bonferroni post hoc test).

[0050] Fig.30 is a series of four images of cortical brain sections of vehicle- or NALL-fed TBI mice stained for TUNEL. Data are presented as mean ± SEM. n=3 for vehicle-fed TBI mice and 4 for NALL-fed TBI mice.

[0051] Fig.31is a bar graph showing area quantification of cortical brain sections of TBI mice stained for TUNEL. Data are presented as mean±SEM. n=3 for vehicle-fed TBI mice and 4 for NALL-fed TBI mice.

[0052] Fig.32 is a bar graph showing cell quantification of TBI mouse cortical brain sections stained for TUNEL. Data are presented as mean ± SEM. n=3 for vehicle-fed TBI mice and 4 for NALL-fed TBI mice.

[0053] Fig.33 Western blot analysis of cortical tissue lysates from sham-operated and TBI mice fed NALL or vehicle was evaluated for the autophagosome marker LC3 and the autophagic cargo protein p62 / SQSTM1.

[0054] Fig.34 is a bar graph showing densitometric analysis of LC3-II relative to actin. Data are presented as mean±SEM. n=5, *p<0.05 (two-way ANOVA with Bonferroni post hoc test).

[0055] Fig.35 is a bar graph showing densitometric analysis of p62 relative to actin. Data are presented as mean±SEM. n=5, *p<0.05 (two-way ANOVA with Bonferroni post hoc test).

[0056] Fig.36 is a bar graph showing the relative mRNA levels of NOS2 in the cortex of sham-operated mice and TBI mice fed with NALL or vehicle. Data are presented as mean ± SEM. n=5, ***p<0.001, *p<0.05 (two-way ANOVA with Bonferroni post hoc test).

[0057] Fig.37 is a bar graph showing the relative mRNA levels of NLRP3 in the cortex of sham-operated mice and TBI mice fed with NALL or vehicle. Data are presented as mean ± SEM. n=5, ***p<0.001, *p<0.05 (two-way ANOVA with Bonferroni post hoc test).

[0058] Fig.38 is a bar graph showing relative mRNA levels of IL-1β in the cortex of sham-operated and TBI mice fed with NALL or vehicle. Data are presented as mean ± SEM. n=5, ***p<0.001, *p<0.05 (two-way ANOVA with Bonferroni post hoc test).

[0059] Fig.39 is a bar graph showing relative mRNA levels of TNF-α in the cortex of sham-operated and TBI mice fed with NALL or vehicle. Data are presented as mean ± SEM. n=5, ***p<0.001, *p<0.05 (two-way ANOVA with Bonferroni post hoc test).

[0060] Fig.40 is a bar graph showing relative mRNA levels of IFN-β in the cortex of sham-operated mice and TBI mice fed with NALL or vehicle. Data are presented as mean ± SEM. n=5, ***p<0.001, *p<0.05 (two-way ANOVA with Bonferroni post hoc test).

[0061] Fig.41 is a bar graph showing the relative mRNA levels of NOX2 in the cortex of sham-operated mice and TBI mice fed with NALL or vehicle. Data are presented as mean ± SEM. n=5, ***p<0.001, *p<0.05 (two-way ANOVA with Bonferroni post hoc test).

[0062] Fig.42 is a bar graph showing the relative mRNA levels of SOCS3 in the cortex of sham-operated mice and TBI mice fed with NALL or vehicle. Data are presented as mean ± SEM. n=5, ***p<0.001, *p<0.05 (two-way ANOVA with Bonferroni post hoc test).

[0063] Fig.43 is a bar graph showing the relative mRNA levels of YM-1 in the cortex of sham-operated mice and TBI mice fed with NALL or vehicle. Data are presented as mean ± SEM. n=5, ***p<0.001, *p<0.05 (two-way ANOVA with Bonferroni post hoc test).

[0064] Fig.44 is a bar graph showing the relative mRNA levels of IL-4ra in the cortex of sham-operated mice and TBI mice fed with NALL or vehicle. Data are presented as mean ± SEM. n=5, ***p<0.001, *p<0.05 (two-way ANOVA with Bonferroni post hoc test).

[0065] Fig.45is a bar graph showing the relative mRNA levels of Arg-1 in the cortex of sham-operated mice and TBI mice fed with NALL or vehicle. Data are presented as mean ± SEM. n=5, ***p<0.001, *p<0.05 (two-way ANOVA with Bonferroni post hoc test).

[0066] Fig.46 Figure 2 shows that untreated Npc1 - / - Mice and Npc1 treated with ADLL, ALL, and ADL - / - Images of cerebellar Purkinje cell loss and microglial activation progressing from the anterior to the posterior lobe in mice. Cerebellum was stained with calbindin-b (Purkinje cells) or CD68 (activated microglia).

[0067] Fig.47 Figure 2 shows that untreated Npc1 - / - Mice and Npc1 treated with ADLL, ALL, and ADL - / - Scatter plot of Purkinje cell survival in mice at 59 days of age.

[0068] Fig.48 Figure 2 shows that untreated Npc1 - / - Mice and Npc1 treated with ADLL, ALL, and ADL - / - Scatter plot of the frequency of CD68-positive activated microglia in mice at 59 days of age. CD68 cell density was measured in the cerebellum. n = 5 animals per group. Mean ± SD, ***p < 0.0009 (one-way ANOVA).

[0069] Fig.49 is a line graph showing motor function (beam walking) results in a controlled cortical impact TBI model in mice treated with NAL.

[0070] Fig.50 is a bar graph showing spatial memory (Y-maze) results in the controlled cortical impact TBI model in mice treated with NAL.

[0071] Fig.51 is a bar graph showing novel object recognition (NOR) results in a controlled cortical impact TBI model in mice treated with NAL. DETAILED DESCRIPTION

[0072] As used herein, a "subject" can be a vertebrate, a mammal, or a domestic animal. Thus, compositions according to the present disclosure can be used to treat any mammal, such as livestock (e.g., horses, cattle, sheep, or pigs), pets (e.g., cats, dogs, rabbits, or guinea pigs), laboratory animals (e.g., mice or rats), or can be used for other veterinary applications. In one embodiment, the subject is a human. "Subject" and "patient" are used interchangeably.

[0073] As used herein, the singular forms "a," "an," and "the" include plural referents.

[0074] The terms "about" and "approximately" mean substantially the same as a referenced number or value, including an acceptable degree of error for the measured quantity given the nature or accuracy of the measurement.

[0075] As used herein, the terms "approximately" and "about" should generally be understood to cover ±20% of a specified amount, frequency or value. Unless otherwise specified, the numerical quantities given herein are approximate, meaning that the terms "about" and "approximately" can be inferred when not explicitly stated.

[0076] As used herein, the terms "administering," "applying," or "administering" refer to (1) providing, administering, administering, and / or prescribing a compound of the present disclosure by or under the direction of a health practitioner or his / her authorized agent, and (2) the patient or he / she himself / herself inserting, taking, or consuming a compound of the present disclosure. Any reference to "leucine," "acetyl-DL-leucine," and "acetyl-L-leucine," or any other compound of the present disclosure includes pharmaceutically acceptable salts thereof, even if not expressly stated.

[0077] Unless otherwise indicated, the term "leucine" refers to L-leucine.

[0078] As mentioned herein, "pharmaceutically acceptable salts" are any salt preparations suitable for pharmaceutical use. Pharmaceutically acceptable salts include, but are not limited to, amine salts such as N,N'-dibenzylethylenediamine, chloroprocaine, choline, ammonia, diethanolamine and other hydroxyalkylamines, ethylenediamine, N-methylglucamine, procaine, N-benzylphenethylamine, 1-p-chlorobenzyl-2-pyrrolidin-1'-ylmethylbenzimidazole, diethylamine and other alkylamines, piperazine, tris(hydroxymethyl)aminomethane, etc.; alkali metal salts such as lithium, potassium, sodium, etc.; alkaline earth metal salts such as barium, calcium, magnesium, etc.; transition metal salts such as zinc, aluminum, etc.; other metal salts such as sodium hydrogen phosphate, disodium phosphate, etc.; inorganic acids such as hydrochloride, sulfate, etc.; and salts of organic acids such as acetate, lactate, malate, tartrate, citrate, ascorbate, succinate, butyrate, valerate, fumarate, etc.

[0079] The compounds of the present disclosure can be prepared according to teachings known in the art and applied to a subject. For example, DL-leucine, L-leucine, D-leucine, L-leucine ethyl ester, acetyl-DL-leucine, acetyl-D-leucine or acetyl-L-leucine can be formulated into a pharmaceutical composition. The pharmaceutical composition can include DL-leucine, L-leucine, D-leucine, L-leucine ethyl ester, acetyl-DL-leucine, acetyl-D-leucine or acetyl-L-leucine and a pharmaceutically acceptable carrier. Mention of the pharmaceutical composition encompasses a single activating agent, that is, DL-leucine, L-leucine, D-leucine, L-leucine ethyl ester, acetyl-DL-leucine, acetyl-D-leucine or acetyl-L-leucine, or in the form of a pharmaceutical composition.

[0080] The pharmaceutical composition may take any of a variety of different forms, depending on how it is to be used. Thus, for example, it may be in the form of a powder, tablet, capsule, liquid, ointment, cream, gel, hydrogel, aerosol, spray, micellar solution, transdermal patch, liposomal suspension or any other suitable form that can be administered to a human or animal in need of treatment.

[0081] As referred to herein, "pharmaceutically acceptable carrier" is any known compound or combination of known compounds known to those skilled in the art to be useful for formulating pharmaceutical compositions, such as excipients, carriers, etc. It should be understood that the carrier of a pharmaceutical composition should be one that is tolerated by the subject to which it is administered.

[0082] In one embodiment, the pharmaceutically acceptable carrier can be solid, and the composition can be in the form of powder or tablet. Solid pharmaceutically acceptable carriers may include, but are not limited to, one or more substances, which may also serve as flavoring agents, buffers, lubricants, stabilizers, solubilizers, suspending agents, wetting agents, emulsifiers, dyes, fillers, glidants, compression aids, inert adhesives, sweeteners, preservatives, dyes, coating agents or tablet disintegrants. The carrier may also be an encapsulating material. In powders, the carrier may be a finely ground solid mixed with a finely ground active agent according to the present invention. In tablets, the active agent may be mixed with a carrier having the necessary compression properties in a suitable ratio and pressed into a desired shape and size. For example, powders and tablets may contain up to 99% of the active agent. Suitable solid carriers include, for example, calcium phosphate, magnesium stearate, talc, sugar, lactose, dextrin, starch, gelatin, cellulose, polyvinyl pyrrolidine, low melting point waxes and ion exchange resins. In another embodiment, the pharmaceutically acceptable carrier may be a gel, and the composition may be in the form of an ointment or the like.

[0083] The carrier may include, but is not limited to, one or more excipients or diluents. Examples of such excipients are gelatin, gum arabic, lactose, microcrystalline cellulose, starch, sodium starch glycolate, calcium hydrogen phosphate, magnesium stearate, talc, colloidal silicon dioxide, and the like.

[0084] In another embodiment, the pharmaceutically acceptable carrier can be a liquid. In one embodiment, the pharmaceutical composition is in the form of a solution. The liquid carrier is used to prepare solutions, suspensions, emulsions, syrups, elixirs and pressurized compositions. The compounds disclosed herein can be dissolved or suspended in a pharmaceutically acceptable liquid carrier (such as water, an organic solvent, a mixture of the two) or in a pharmaceutically acceptable oil or fat. The liquid carrier may contain other suitable pharmaceutical additives, such as solubilizers, emulsifiers, buffers, preservatives, sweeteners, flavoring agents, suspending agents, thickeners, colorants, viscosity modifiers, stabilizers or osmotic pressure regulators. Suitable examples of liquid carriers for oral and parenteral administration include water (partially containing the above-mentioned additives, such as cellulose derivatives, such as sodium carboxymethylcellulose solution), alcohols (including monohydric alcohols and polyols, such as diols) and derivatives thereof, and oils (such as fractionated coconut oil and peanut oil). For parenteral administration, the carrier may also be a grease, such as ethyl oleate and isopropyl myristate. Sterile liquid carriers can be used in sterile liquid form compositions for parenteral administration. The liquid carrier for pressurized compositions can be halogenated hydrocarbon or other pharmaceutically acceptable propellant.

[0085] Liquid pharmaceutical compositions (which are sterile solutions or suspensions) can be utilized by, for example, intramuscular, intrathecal, epidural, intraperitoneal, intravenous, and subcutaneous injection. The active agent can be prepared as a sterile solid composition that can be dissolved or suspended using sterile water, saline or other suitable sterile injectable media when administered.

[0086] The composition can be orally administered in the form of a sterile solution or suspension containing other solutes or suspending agents (e.g., saline or glucose sufficient to make the solution isotonic), bile salts, gum arabic, gelatin, sorbitan monooleate, polysorbate 80 (oleate copolymerized with sorbitol and its anhydride and ethylene oxide), etc. The composition can also be orally administered in the form of a liquid or solid composition. Compositions suitable for oral administration include solid forms (such as pills, capsules, granules, tablets and powders) and liquid forms (such as solutions, syrups, elixirs and suspensions). Forms that can be used for parenteral administration include sterile solutions, emulsions and suspensions.

[0087] The composition may alternatively be administered by inhalation (e.g., intranasally). The composition may also be formulated for topical use. For example, a cream or ointment may be applied to the skin.

[0088] The compounds of the present disclosure may be incorporated into a slow or delayed release device. Such devices may, for example, be inserted on or under the skin, and the drug may be released over weeks or even months. Such devices may be advantageous when long-term treatment with the compounds of the present disclosure used according to the present disclosure is required and frequent administration (e.g., at least daily administration) may be required.

[0089] In one embodiment, the pharmaceutical composition is a solid oral dosage form, such as a tablet. In a tablet, the active agent may be mixed with a vehicle having the necessary compression properties, such as a pharmaceutically acceptable carrier, in a suitable ratio and compressed into a desired shape and size. The tablet may contain up to 99% by weight of the active agent.

[0090] Pharmaceutical compositions in solid oral dosage forms such as tablets can be prepared by any method known in the pharmaceutical art. Pharmaceutical compositions are generally prepared by mixing the active agent with a conventional pharmaceutically acceptable carrier.

[0091] Tablets may be formulated as known in the art. For example, Wheat starch, pregelatinized corn (maize) starch, calcium carbonate and magnesium stearate are included as excipients. For example, the same or similar excipients can be used in the present disclosure.

[0092] Each 700mg The composition of the tablets is as follows: 500 mg acetyl-DL-leucine, 88 mg wheat starch, 88 mg pregelatinized corn (maize) starch, 13 mg calcium carbonate, and 11 mg magnesium stearate. For example, the same tablets can be used in the methods of the present disclosure.

[0093] As described above, the compounds of the present disclosure can be formulated into pharmaceutical compositions in a variety of different forms and administered. For example, the compounds of the present disclosure can be formulated into pharmaceutical compositions to facilitate their delivery across the blood-brain barrier. As other examples, the compounds of the present disclosure can be formulated into pharmaceutical compositions for bypassing the blood-brain barrier. Preparations that facilitate delivery across the blood-brain barrier or are suitable for delivery in a manner that bypasses the blood-brain barrier can be used to prepare and administer leucine (unacetylated) as described herein.

[0094] In one embodiment, for example, a pharmaceutical composition comprising acetyl-L-leucine or a salt thereof is formulated for nano-delivery, such as a colloidal drug-carrier system. Suitable examples include, but are not limited to, liposomes, nanoparticles (e.g., polymers, lipids, and inorganic nanoparticles), nanogels, dendrimers, micelles, nanoemulsions, polymer vesicles, exosomes, and quantum dots. See, e.g., Patel et al., "Crossing the Blood-Brain Barrier: Recent Advances in Drug Delivery to the Brain," CNSDrugs 31: 109-133 (2017); Kabanov et al., "New Technologies for Drug Delivery across the Blood Brain Barrier," Curr Pharm Des., 10(12): 1355-1363 (2004); Cheng et al., "Highly Stabilized Curcumin Nanoparticles Tested in an In Vitro Blood–Brain Barrier Model and in Alzheimer's Disease Tg2576 Mice," The AAPS Journal, Vol. 15, No. 2, pp. 324-336 (2013); et al. "Production of L-Leucine Nanoparticles under Various Conditions Using an Aerosol Flow Reactor Method," Journal of Nanomaterials, Vol. 2008, Article ID 680897 (2008).

[0095] In one embodiment, for example, a pharmaceutical composition comprising acetyl-L-leucine or a salt thereof is formulated for direct delivery to the central nervous system (CNS), such as by injection or infusion. Preparations and methods for direct delivery to the CNS are known in the art. See, for example, U.S. Patent No. 9,283,181. Examples of such administration include, but are not limited to, intranasal, intraventricular, intrathecal, intracranial, and delivery by nasal mucosal transplantation.

[0096] In one embodiment, the pharmaceutical composition is formulated for intranasal delivery (and is administered by intranasal delivery). See, for example, Hanson et al., "Intranasal delivery bypasses the blood-brain barrier to target therapeutic agents to the central nervous system and treat neurodegenerative disease," BMC Neurosci.9 (Suppl 3): S5 (2008). In one embodiment, the pharmaceutical composition is formulated for delivery via a nasal mucosal graft (and is administered by delivery via a nasal mucosal graft). In one embodiment, the pharmaceutical composition is formulated for intraventricular injection or infusion (and is administered by intraventricular injection or infusion). In another embodiment, the pharmaceutical composition is formulated for intrathecal intracisternal injection or infusion (and is administered by intrathecal intracisternal injection or infusion). In one embodiment, the pharmaceutical composition is formulated for intrathecal lumbar injection or infusion (and is administered by intrathecal lumbar injection or infusion).

[0097] Various techniques can be used, including but not limited to injection through a burr hole in the skull or a cisternal or lumbar puncture as known in the art. Various devices, whether internal (e.g., implanted) or external, can be used for delivery, as known in the art, such as pumps, catheters, reservoirs, etc. In one embodiment, the administration interval is once every two weeks.

[0098] In one embodiment, the administration interval is once a month. In one embodiment, the administration interval is once every two months. In one embodiment, the administration interval is twice a month. In one embodiment, the administration interval is once a week. In one embodiment, the administration interval is twice or several times a week. In one embodiment, the administration interval is once a day. In one embodiment, administration is continuous, such as continuous infusion.

[0099] In one embodiment, the dose or equivalent amount of a compound of the present disclosure may be adjusted to account for its direct delivery to the CNS or its delivery across the blood-brain barrier.

[0100] The present disclosure describes compounds of the present disclosure, including pharmaceutical compositions thereof, for use in treating various diseases, conditions, disorders and syndromes in a subject in need thereof.

[0101] As used herein, "traumatic brain injury" refers to any injury to the brain that has an initial phase consisting of at least one head trauma event (i.e., an object entering the brain and / or a load or force on the brain that causes the brain to move rapidly and / or unnaturally within the subject's skull) and a subsequent second phase (including physiological, cellular and / or molecular disturbances caused by head trauma). Examples of head trauma include, for example, objects that penetrate the skull, such as bullets, arrows, and other objects that pass through the skull and enter the brain; impact loads or forces applied to the patient's head or other parts of the body; trauma caused by surgery; shock caused by explosions; tremors caused by non-explosive means such as sports injuries, car accidents, building collapses, and earthquakes. The results of TBI can be of various types, and all forms of TBI are within the scope of the present disclosure. For example, the severity of TBI can range from a brief change in mental state or consciousness to a long period of unconsciousness or memory loss. In one embodiment, TBI is caused by a concussion. In one embodiment, TBI is caused by a contusion. In one embodiment, TBI is caused by an open head injury. In another embodiment, TBI is caused by a closed head injury. In one embodiment, the TBI is caused by focal injury. In another embodiment, the TBI is caused by diffuse injury. In one embodiment, the TBI is in the form of mild TBI. In another embodiment, the TBI is in the form of moderate TBI. In another embodiment, the TBI is in the form of severe TBI.

[0102] As used herein, a "subject in need thereof" can be any subject having a disease, disorder, condition, or syndrome, such as any of the diseases, disorders, conditions, or syndromes provided in any of Tables 1-17. The subject may or may not have been diagnosed with the disease, disorder, condition, or syndrome. For example, a subject may not have been diagnosed (clinically or otherwise) with TBI, but may have one or more symptomatic TBIs. The subject may also have biochemical or other similarly identifiable markers of TBI.

[0103] A "therapeutically effective amount" of a compound of the present disclosure is any amount that is required to produce the desired effect when administered to a subject, which for the present disclosure may be therapeutic and / or prophylactic. The dosage may be determined according to various parameters, such as the specific compound of the present disclosure used; the age, weight, and condition of the patient to be treated; the route of administration; and the desired regimen. The physician will be able to determine the route of administration and dosage required for any particular patient. For example, the daily dosage may be about 10 to about 225 mg / kg, about 10 to about 150 mg / kg, or about 10 to about 100 mg / kg body weight.

[0104] As used herein, "treating" or "treatment" refers to successfully preventing, suppressing or ameliorating a disease, disorder, condition or syndrome, such as a disease, disorder, condition or syndrome provided in any one of Tables 1-17, and / or preventing, suppressing or ameliorating any signs of any one or more symptoms of a disease, disorder, condition or syndrome, such as a disease, disorder, condition or syndrome provided in any one of Tables 1-17, in a subject, including any objective or subjective parameter, such as alleviating; alleviating; preventing, reducing, inhibiting or eliminating one or more symptoms; making the disease, disorder, condition or syndrome more tolerable to the subject; slowing the progression of the disease, disorder, condition or syndrome; or improving the physical or mental health of a subject in need thereof.

[0105] The terms "treating" or "treatment" also encompass, for example, inducing inhibition, regression, or arrest of a disease, disorder, condition, or syndrome. For example, treatment of a patient or subject in need of treatment for TBI includes alleviating the symptoms of TBI in the subject, inducing a clinical response, inhibiting or alleviating the progression of TBI, or inhibiting or alleviating complications of TBI.

[0106] Preventing, arresting or ameliorating damage or pathology of a disease, condition, disorder or syndrome, such as preventing, reducing, inhibiting or eliminating one or more symptoms of a disease, condition, disorder or syndrome can be based on objective and / or subjective parameters, including, for example, the results of a physical examination, a neurological examination and / or a psychiatric assessment. The success of treatment of certain diseases listed in Tables 1-17 (e.g., TBI) can be measured or assessed by, for example, comparing the severity of the disease before starting treatment with the compounds of the present disclosure (e.g., objective and / or subjective parameters of TBI) with the severity of the disease after starting treatment with the compounds of the present disclosure (e.g., objective and / or subjective parameters of TBI). For example, the severity of TBI can be assessed using a scale, index, rating or score. In one embodiment, the treatment described herein improves this assessment from a value or degree characteristic of a symptomatic subject to a value or degree characteristic of an asymptomatic subject. In one embodiment, the treatment described herein improves this assessment compared to a baseline. The baseline can be, for example, the condition of the subject before starting any treatment for the disease (e.g., TBI), or before starting treatment of the disease with the compounds of the present disclosure. Alternatively, the baseline can be, for example, the condition of the subject after a period of treatment of the disease. In one embodiment, treatment with a compound of the present disclosure as described herein improves the subject's assessment (e.g., a scale, index, rating, or score of an objective and / or subjective parameter) by at least 10%, at least 20%, at least 30%, at least 40%, or at least 50% compared to baseline. In one embodiment, the assessment is improved by at least 60%, at least 70%, at least 80%, at least 90%, or 100%.

[0107] For example, in one embodiment, the severity of a patient's disease (e.g., TBI) can be quantified by the Glasgow Coma Scale (GCS). The GCS is a cumulative score of three examination areas: eyes, language, and motor function. For eye examination, the patient is graded from 1 to 4 as follows: 1-no eyes open to any stimulus, 2-open eyes only when responding to pain, 3-open eyes when speaking, and 4-eyes open automatically. For language examination, the patient is graded from 1 to 5 as follows: 1-no language response, 2-unintelligible sounds, 3-inappropriate words, 4-confusion, and 5-direction. For motor examination, the patient is graded from 1 to 6 as follows: 1-no motor response, 2-pain extension, 3-pain abnormal flexion, 4-pain withdrawal, 5-pain positioning, and 6-obeying commands. The GCS score is the sum of the three scores received for eyes, language, and motor response. In general, TBI is classified as mild, moderate, or severe based on the Glasgow Coma Scale as follows: mild: GCS>13, moderate: GCS 9-12, and severe: GCS<8. The level of recovery of TBI patients can be quantified by the Glasgow Outcome Scale (GOS). The Glasgow Outcome Scale is a 5-point score: 1-death, 2-vegetative state, 3-severe disability, 4-moderate disability, and 5-good recovery. The GOS is generally divided into "favorable" outcomes (moderate disability and good recovery) and "unfavorable" outcomes (death, vegetative state, and severe disability).

[0108] "Symptoms" of a disease, disorder, condition, or syndrome include any clinical or laboratory manifestation associated with the disease, disorder, condition, or syndrome, and are not limited to what can be felt or observed by a subject. For example, symptoms of certain diseases listed in Tables 1-17' (e.g., TBI) include, but are not limited to, loss of consciousness, confusion, restlessness, agitation, disorientation, memory loss, headache, dizziness, vertigo, blurred vision or eyestrain, fatigue or drowsiness, changes in sleep patterns, behavioral or mood swings, memory, concentration, attention and / or thinking problems, repeated nausea or vomiting, convulsions or seizures, inability to wake from sleep, slurred speech, weakness or numbness in the limbs, and loss of coordination.

[0109] In one embodiment, the compounds of the present disclosure may be administered, for example, in a dosage range of about 500 mg to about 30 g per day, or in a dosage range of about 500 mg to about 15 g per day, such as in a dosage range of about 1.5 g to about 10 g per day, optionally by a solid oral or liquid oral route. The compounds of the present disclosure may be administered, for example, in a dosage range of about 1.5 g to about 10 g per day. The dosage is administered at a dosage of 1.5 g to 2 g per day for adults, taken in two divided doses of 3-4 tablets in the morning and evening.

[0110] If a single enantiomer is administered, i.e., L-leucine, acetyl-D-leucine, or acetyl-L-leucine, the dosage may be reduced accordingly. For example, if only acetyl-L-leucine is administered or if only acetyl-D-leucine is administered, the dosage may be in the range of about 250 mg to about 15 g per day, in the range of about 250 mg to about 10 g per day, or in the range of about 250 mg to about 5 g per day, such as about 0.75 g to about 5 g per day.

[0111] In one embodiment, the administered dose is in the range of about 1 g to about 30 g per day, about 1 g to about 15 g per day, about 1 g to about 10 g per day, or about 1.5 g to about 7 g per day, 15.1 g to about 30 g per day, 16 g to about 30 g per day, 17 g to about 30 g per day, 18 g to about 30 g per day, 19 g to about 30 g per day, or 20 g to about 30 g per day. It can be about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 g to about 15 g per day. It can be about 2, 3, 4, 5, 6, 7, 8, or 9 g to about 10 g per day. It can be 15.1, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 25, 27, 28, or 29 g to about 30 g per day. It can be more than about 1.5g per day, but less than about 15, 14, 13, 12, 11, 10, 9, 8, 7, 6 or 5g per day. In one embodiment, the dosage is in the range of about 4g to about 6g per day. In one embodiment, the dosage is in the range of about 4g to about 5g per day. In one embodiment, the dosage is about 4.5g per day. In one embodiment, the dosage is about 5g per day. In one embodiment, the dosage is about 1g per day, about 2g per day, about 3g per day, about 4g per day, about 5g per day, about 6g per day, about 7g per day, about 8g per day, about 9g per day, about 10g per day, about 11g per day, about 12g per day, about 13g per day, about 14g per day or about 15g per day. In another embodiment, the dosage is about 16g per day, about 17g per day, about 18g per day, about 19g per day or about 20g per day. In another embodiment, the dosage is about 21 g per day, about 22 g per day, about 23 g per day, about 24 g per day, about 25 g per day, about 26 g per day, about 27 g per day, about 28 g per day, about 29 g per day, or about 30 g per day. In one embodiment, these dosages are administered in solid oral dosage forms, particularly tablets. In another embodiment, these dosages are for acetyl-leucine in its racemic form. When there is an enantiomeric excess, the dosage of acetyl-leucine may be lower, for example, about 50% lower. Therefore, when halved, the above dosage ranges are also clearly included in the present disclosure.

[0112] In one embodiment, the total daily dose can be distributed in multiple administrations, i.e., administration can occur twice or more times a day to reach the total daily dose. For example, the number of tablets required to provide the total daily dose of the compound of the present disclosure can be divided into two administrations (e.g., morning and evening) or three administrations (e.g., morning, noon and evening). Each dose can be appropriately administered with or without food. For example, acetyl-L-leucine or acetyl-DL-leucine can be administered about 1 or about 2 hours before a meal, such as at least about 20 minutes, at least about 30 minutes, at least about 40 minutes or at least about 1 hour before a meal, or can be administered about 1, about 2 or about 3 hours after a meal, such as waiting for at least about 20 minutes, at least about 30 minutes, at least about 1 hour, at least about 1.5 hours, at least about 2 hours or at least about 2.5 hours after a meal. For example, the total daily dose of 4.5g acetyl-DL-leucine can be taken as three tablets before breakfast, during breakfast or after breakfast. (or equivalent) tablets, another three tablets before, at, or after lunch, and another three tablets before, at, or after dinner.

[0113] In one embodiment, with respect to TBI, administration of the compounds of the present disclosure according to the present disclosure begins at or around (e.g., within 24 hours) the time of head trauma. In one embodiment, administration begins after a period of time (e.g., days, weeks, months, or even years) after the head trauma. In one embodiment, administration begins after the subject is diagnosed (clinically or otherwise) with TBI. In one embodiment, administration begins after the subject is diagnosed with TBI using the Glasgow Coma Scale (GCS). In one embodiment, the subject's GCS score is 13-15. In another embodiment, the subject's GCS score is 9-12. In another embodiment, the subject's GCS score is 3-8. In one embodiment, administration begins before head trauma (or before subsequent head trauma if one or more head traumas have occurred). In one embodiment, administration begins when, before, after, or after the subject is found to have a biochemical or other similar identifiable marker of TBI. In one embodiment, administration begins when, before, after, or after the subject experiences one or more symptoms of TBI.

[0114] The treatment duration of any disease listed in Table 1-17 can be, for example, about seven days or longer, about two weeks or longer, about three weeks or longer, about one month or longer, about six weeks or longer, about seven weeks or longer, or about two months or longer. In one embodiment, the treatment duration is about three months or longer, about four months or longer, about five months or longer, or about six months or longer. The treatment duration can be about 1 year or longer, about 2 years or longer, about 4 years or longer, about 5 years or longer, or about 10 years or longer. The treatment duration can be the survival period of the experimenter.

[0115] The present disclosure contemplates and encompasses any and all combinations of dosage forms, dosages, dosing schedules, and durations of treatment. In one embodiment, the dosage is about 4 g to about 10 g per day, administered once, twice, or three times per day, for a duration of treatment of about two months or longer. In another embodiment, the dosage is more than 4 g but not more than 5 g per day, administered once, twice, or three times per day, for a duration of treatment of about six months or longer. The dosage form may be a solid oral dosage form, particularly a tablet.

[0116] The compounds of the present disclosure can be used as a monotherapy (e.g., an active agent used alone) to treat a disease, disorder, condition, or syndrome in a subject, such as a disease, disorder, condition, or syndrome provided in any one of Tables 1 to 17. Alternatively, the compounds of the present disclosure can be used as an adjunct to or in combination with other known therapies for treating a disease, disorder, condition, or syndrome, such as a disease, disorder, condition, or syndrome provided in any one of Tables 1 to 17.

[0117] Also disclosed is a kit for treating a disease, disorder, condition or syndrome in a subject, such as a disease, disorder, condition or syndrome provided in any one of Tables 1-17, comprising a device for diagnosing or prognosing the disease, disorder, condition or syndrome and a compound of the present disclosure.

[0118] The kit may also include a buffer or aqueous solution.The kit may also include instructions for using the compounds of the disclosure according to the methods of the disclosure.

[0119] All features described herein (including any accompanying claims, abstract and drawings) and / or all steps of any method so disclosed may be combined in any combination with any of the above aspects, except combinations where at least some of such features and / or steps are mutually exclusive.

[0120] The phrase "a disease, condition, disorder or syndrome in which modulation of one or more proinflammatory mediators provides a benefit" and the like refers to (i): a disease, condition, disorder or syndrome in which one or more proinflammatory mediators are important or necessary, for example, for the onset, progression or expression of the disease, condition, disorder or syndrome, or; (ii) a disease, condition, disorder or syndrome known to be responsive to modulation of proinflammatory mediators. Examples of such diseases include, but are not limited to, neuroinflammatory diseases, nervous system diseases, autoimmune diseases, neurodegenerative protein diseases, psychiatric disorders (e.g., depression) and mitochondrial diseases. Proinflammatory mediators are well known in the art. See, for example, "Really Essential Medical Immunology," 2nd edition, Rabson, Roitt, Delves (Blackwell, 2005), exemplary non-limiting proinflammatory mediators include cytokines, such as interleukin (IL) -1β, IL-8, tumor necrosis factor α (TNF-α), IL-6 and IL-12, chemokines and growth factors.

[0121] In one embodiment, the disclosure provides a method of treating a disease, disorder, condition or syndrome in a subject in need thereof, or treating a symptom of a disease, disorder, condition or syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the disclosure, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 1.

[0122] Table 1

[0123]

[0124]

[0125]

[0126]

[0127] In another embodiment, the disclosure provides a method of treating a disease, disorder, condition or syndrome in a subject in need thereof, or treating a symptom of a disease, disorder, condition or syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the disclosure, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 2.

[0128] Table 2

[0129]

[0130]

[0131] In one embodiment, the disclosure provides a method of treating a disease, disorder, condition or syndrome in a subject in need thereof, or treating a symptom of a disease, disorder, condition or syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the disclosure, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 3.

[0132] Table 3

[0133]

[0134]

[0135] In another embodiment, the disclosure provides a method of treating a neuroinflammatory disease, disorder, condition or syndrome in a subject in need thereof, or treating a symptom of a neuroinflammatory disease, disorder, condition or syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the disclosure.

[0136] In another embodiment, the disclosure provides a method of treating a disease, disorder, condition or syndrome in a subject in need thereof, or treating a symptom of a disease, disorder, condition or syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the disclosure, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 4.

[0137] Table 4

[0138]

[0139]

[0140] In another embodiment, the disclosure provides a method of treating a disease, disorder, condition or syndrome in a subject in need thereof, or treating a symptom of a disease, disorder, condition or syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the disclosure, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 5. In another embodiment, the disclosure provides a method of treating traumatic brain injury.

[0141] Table 5

[0142]

[0143]

[0144] In another embodiment, the disclosure provides a method of treating a neurological disease, disorder, condition or syndrome in a subject in need thereof, or treating a symptom of a neurological disease, disorder, condition or syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the disclosure.

[0145] In another embodiment, the disclosure provides a method of treating a disease, disorder, condition or syndrome in a subject in need thereof, or treating a symptom of a disease, disorder, condition or syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the disclosure, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 6.

[0146] Table 6

[0147]

[0148]

[0149]

[0150]

[0151] In another embodiment, the disclosure provides a method of treating a disease, disorder, condition or syndrome in a subject in need thereof, or treating a symptom of a disease, disorder, condition or syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the disclosure, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 7.

[0152] Table 7

[0153]

[0154]

[0155]

[0156] In another embodiment, the disclosure provides a method of treating an autoimmune disease, disorder, condition or syndrome in a subject in need thereof, or treating a symptom of an autoimmune disease, disorder, condition or syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the disclosure.

[0157] In another embodiment, the disclosure provides a method of treating a disease, disorder, condition or syndrome in a subject in need thereof, or treating a symptom of a disease, disorder, condition or syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the disclosure, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 8.

[0158] Table 8

[0159] Autoimmune thrombocytopenia diabetes Idiopathic thrombocytopenic purpura (ITP) IgA nephropathy IgM polyneuropathy Inflammatory bowel disease Juvenile rheumatoid arthritis Lupus nephritis Multiple sclerosis Myasthenia gravis psoriasis Raynaud's syndrome Rheumatoid arthritis Sjögren's syndrome and glomerulonephritis Systemic lupus erythematosus (SLE) Thrombotic thrombocytopenic purpura (TTP) Vasculitis Wegener's disease

[0160] In another embodiment, the disclosure provides a method of treating a disease, disorder, condition or syndrome in a subject in need thereof, or treating a symptom of a disease, disorder, condition or syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the disclosure, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 9.

[0161] Table 9

[0162] Autoimmune thrombocytopenia Idiopathic thrombocytopenic purpura (ITP) IgA nephropathy IgM polyneuropathy Inflammatory bowel disease Lupus nephritis Myasthenia gravis psoriasis Raynaud's syndrome Rheumatoid arthritis Sjögren's syndrome and glomerulonephritis Systemic lupus erythematosus (SLE) Thrombotic thrombocytopenic purpura (TTP) Wegener's disease

[0163] In another embodiment, the present disclosure provides a method of treating a neurodegenerative protein disease, disorder, condition or syndrome in a subject in need thereof, or treating a symptom of a neurodegenerative protein disease, disorder, condition or syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the present disclosure.

[0164] In another embodiment, the disclosure provides a method of treating a disease, disorder, condition or syndrome in a subject in need thereof, or treating a symptom of a disease, disorder, condition or syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the disclosure, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 10.

[0165] Table 10

[0166]

[0167]

[0168]

[0169]

[0170] In another embodiment, the disclosure provides a method of treating a disease, disorder, condition or syndrome in a subject in need thereof, or treating a symptom of a disease, disorder, condition or syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the disclosure, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 11.

[0171] Table 11

[0172]

[0173]

[0174]

[0175] In another embodiment, the disclosure provides a method of treating a psychiatric disease, disorder, condition or syndrome, or treating a symptom of a psychiatric disease, disorder, condition or syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the disclosure.

[0176] In another embodiment, the disclosure provides a method of treating a disease, disorder, condition or syndrome in a subject in need thereof, or treating a symptom of a disease, disorder, condition or syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the disclosure, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 12.

[0177] Table 12

[0178] depression Generalized anxiety disorder Major depressive disorder Obsessive-compulsive disorder Panic disorder Post-traumatic stress Social phobia

[0179] In another embodiment, the disclosure provides a method of treating a disease, disorder, condition or syndrome in a subject in need thereof, or treating a symptom of a disease, disorder, condition or syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the disclosure, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 13.

[0180] Table 13

[0181] Generalized anxiety disorder Obsessive-compulsive disorder Panic disorder Post-traumatic stress Social phobia

[0182] In another embodiment, the present disclosure provides a method of treating a mitochondrial disease, disorder, condition or syndrome in a subject in need thereof, or treating a symptom of a mitochondrial disease, disorder, condition or syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the present disclosure.

[0183] In another embodiment, the disclosure provides a method of treating a disease, disorder, condition or syndrome in a subject in need thereof, or treating a symptom of a disease, disorder, condition or syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the disclosure, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 14.

[0184] Table 14

[0185]

[0186] In another embodiment, the disclosure provides a method of treating a disease, disorder, condition or syndrome in a subject in need thereof, or treating a symptom of a disease, disorder, condition or syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the disclosure, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 15.

[0187] Table 15

[0188]

[0189] In another embodiment, the disclosure provides a method of treating a disease, disorder, condition or syndrome in a subject in need thereof, or treating a symptom of a disease, disorder, condition or syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the disclosure, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 16.

[0190] Table 16

[0191] Spinal muscular atrophy Friedreich's ataxia Charcot-Marie-Tooth Disease Type 2K Wilson disease

[0192] In another embodiment, the disclosure provides a method of treating a disease, disorder, condition or syndrome in a subject in need thereof, or treating a symptom of a disease, disorder, condition or syndrome in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of the disclosure, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 17.

[0193] Table 17

[0194] Acoustic neuroma Anencephaly Batten's disease catatonia Cerebral palsy Chorea in children and adults Chromosome 8P Churg-Struass disease Conus and cauda equina syndrome Dopamine-responsive dystonia Dropped foot Facioscapulohumeral dystrophy Infant Scoliosis Kennedy disease Klippel-Feil syndrome Labyrinthitis Lambert-Eaton myasthenic syndrome (LEMS) Lin-Gordon syndrome Lesch-Nyhan disease Menkes disease Moyamoya disease Olivopontocerebellar atrophy (OPCA) Pantothenate kinase-associated neurodegeneration Guillain-Barré syndrome in children Stiff-person syndrome West disease Emory Dreifuss Muscular Dystrophy

[0195] In another embodiment, the present disclosure provides a method for treating a disease, condition, illness or syndrome (e.g., a disease, condition, illness or syndrome provided in any one of Tables 1-17) or treating a disease, condition, illness or syndrome of a subject in need thereof, the method comprising administering to the subject about 1 g to about 30 g of a compound of the present disclosure per day. In another embodiment, about 2 g to about 15 g of a compound of the present disclosure is administered to the subject per day. In another embodiment, about 3 g to about 10 g of a compound of the present disclosure is administered to the subject per day. In another embodiment, about 4 g to about 8 g of a compound of the present disclosure is administered to the subject per day. In another embodiment, about 4 g to about 5 g of a compound of the present disclosure is administered to the subject per day. In another embodiment, about 15.1 g to about 30 g of a compound of the present disclosure is administered to the subject per day. In another embodiment, about 5 g of a compound of the present disclosure is administered to the subject per day.

[0196] In another embodiment, the present disclosure provides a method of treating a disease, condition, illness, or syndrome (e.g., a disease, condition, illness, or syndrome provided in any one of Tables 1-17) in a subject in need thereof, or treating a symptom of a disease, condition, illness, or syndrome in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of DL-leucine or a pharmaceutically acceptable salt thereof. In another embodiment, a therapeutically effective amount of DL-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the disease, condition, illness, or syndrome. In another embodiment, a therapeutically effective amount of DL-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the symptom of the disease, condition, illness, or syndrome.

[0197] In another embodiment, the present disclosure provides a method for treating a disease, condition, illness or syndrome (e.g., a disease, condition, illness or syndrome provided in any one of Tables 1-17) in a subject in need thereof, or treating a symptom of a disease, condition, illness or syndrome in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of L-leucine or a pharmaceutically acceptable salt thereof. In another embodiment, a therapeutically effective amount of L-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the disease, condition, illness or syndrome. In another embodiment, a therapeutically effective amount of L-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the symptom of the disease, condition, illness or syndrome.

[0198] In another embodiment, the present disclosure provides a method of treating a disease, condition, illness, or syndrome (e.g., a disease, condition, illness, or syndrome provided in any one of Tables 1-17) in a subject in need thereof, or treating a symptom of a disease, condition, illness, or syndrome in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of D-leucine or a pharmaceutically acceptable salt thereof. In another embodiment, a therapeutically effective amount of D-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the disease, condition, illness, or syndrome. In another embodiment, a therapeutically effective amount of D-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the symptom of the disease, condition, illness, or syndrome.

[0199] In another embodiment, the present disclosure provides a method of treating a disease, condition, illness, or syndrome (e.g., a disease, condition, illness, or syndrome provided in any one of Tables 1-17) in a subject in need thereof, or treating a symptom of a disease, condition, illness, or syndrome in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of L-leucine ethyl ester or a pharmaceutically acceptable salt thereof. In another embodiment, a therapeutically effective amount of L-leucine ethyl ester or a pharmaceutically acceptable salt thereof is administered to the subject to treat the disease, condition, illness, or syndrome. In another embodiment, a therapeutically effective amount of L-leucine ethyl ester or a pharmaceutically acceptable salt thereof is administered to the subject to treat the symptom of the disease, condition, illness, or syndrome.

[0200] In another embodiment, the present disclosure provides a method of treating a disease, disorder, condition, or syndrome (e.g., a disease, disorder, condition, or syndrome provided in any one of Tables 1-17) in a subject in need thereof, or treating a symptom of a disease, disorder, condition, or syndrome in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of acetyl-DL-leucine or a pharmaceutically acceptable salt thereof. In another embodiment, a therapeutically effective amount of acetyl-DL-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the disease, disorder, condition, or syndrome. In another embodiment, a therapeutically effective amount of acetyl-DL-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the disease, disorder, condition, or syndrome.

[0201] In another embodiment, the present disclosure provides a method of treating a disease, disorder, condition, or syndrome (e.g., a disease, disorder, condition, or syndrome provided in any one of Tables 1-17) in a subject in need thereof, or treating a symptom of a disease, disorder, condition, or syndrome in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of acetyl-D-leucine or a pharmaceutically acceptable salt thereof. In another embodiment, a therapeutically effective amount of acetyl-D-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the disease, disorder, condition, or syndrome. In another embodiment, a therapeutically effective amount of acetyl-D-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the disease, disorder, condition, or syndrome.

[0202] In another embodiment, the present disclosure provides a method of treating a disease, disorder, condition, or syndrome (e.g., a disease, disorder, condition, or syndrome provided in any one of Tables 1-17) in a subject in need thereof, or treating a symptom of a disease, disorder, condition, or syndrome in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of acetyl-L-leucine or a pharmaceutically acceptable salt thereof. In another embodiment, a therapeutically effective amount of acetyl-L-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the disease, disorder, condition, or syndrome. In another embodiment, a therapeutically effective amount of acetyl-L-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the disease, disorder, condition, or syndrome.

[0203] In another aspect, the present disclosure provides a method of regulating the expression of one or more proinflammatory mediators (e.g., proinflammatory cytokines and proinflammatory chemokines) in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of the present disclosure. In another embodiment, the proinflammatory mediator is any one or more of NOS2, IL-18, IFNb, IL-1β, TNFα, NOX2, NLRP3, SOCS3, ARG1, IL-10, IL-4ra or YM1.

[0204] In another embodiment, administration of the compounds of the present disclosure reduces or inhibits neuroinflammation in a subject in need thereof. In another embodiment, the reduced or inhibited neuroinflammation includes reduction or inhibition of excessive proinflammatory cytokines. In another embodiment, neuroinflammation is inhibited or reduced compared to neuroinflammation in a subject with a disease, disorder, illness or syndrome, such as a disease, disorder, illness or syndrome (e.g., TBI) provided in any one of Tables 1-17, to which a compound of the present disclosure is not administered.

[0205] In another embodiment, the administration of the compounds of the present disclosure regulates the elevated levels of microtubule-associated protein Tau and / or microtubule-associated protein 1A / 1B-light chain 3-phosphatidylethanolamine conjugate (LC3-II) in subjects in need of treatment for TBI. In one embodiment, the elevated levels of Tau protein and / or LC3-II are regulated compared to elevated levels of Tau protein and / or LC3-II in subjects with a disease, disorder, condition or syndrome, such as any one of Tables 1-17 provided (e.g., TBI), to which the compounds of the present disclosure are not administered.

[0206] In another embodiment, administration of a Compound of the Disclosure is effective to improve recovery in a subject in need thereof. In another embodiment, recovery is improved compared to recovery in a subject with a disease, disorder, condition, or syndrome, such as a disease, disorder, condition, or syndrome provided in any one of Tables 1-17 (e.g., TBI), to whom a Compound of the Disclosure is not administered.

[0207] In another embodiment, recovery is assessed using the Glasgow Outcome Scale (GOS). In one embodiment, administration of leucine, acetyl-leucine, or any other compound of the present disclosure, or a pharmaceutically acceptable salt thereof, is effective to increase the percentage of subjects with a "favorable" outcome according to the GOS by at least 10%, at least 20%, at least 30%, at least 50%, or at least 70% compared to baseline, such as compared to subjects with TBI who were not administered leucine, acetyl-leucine, or a pharmaceutically acceptable salt thereof. In one embodiment, the subject is assessed using the GOS three months after the injury is inflicted. In another embodiment, the subject is assessed using the GOS six months after the injury is inflicted. In another embodiment, the subject is assessed using the GOS 12 months or more after the injury is inflicted.

[0208] In another embodiment, the administration of the compounds of the present disclosure effectively improves the cognitive function of a subject in need. In another embodiment, compared with the cognitive function of a subject with a disease, condition, illness or syndrome, such as a disease, condition, illness or syndrome (e.g., TBI) provided in any one of Tables 1-17 to which the compounds of the present disclosure are not administered, the cognitive function is improved. In one embodiment, the cognitive function is memory. In one embodiment, memory is long-term memory. In another embodiment, the cognitive function is learning. In another embodiment, the cognitive function is spatial navigation.

[0209] In another embodiment, administration of a compound of the present disclosure reduces or inhibits at least one symptom of a disease, disorder, condition, or syndrome, such as a disease, disorder, condition, or syndrome provided in any one of Tables 1-17 (e.g., TBI), in a subject in need thereof. In another embodiment, at least one symptom is reduced or inhibited as compared to a symptom of a subject with a disease, disorder, condition, or syndrome, such as a disease, disorder, condition, or syndrome provided in any one of Tables 1-17 (e.g., TBI) who is not administered a compound of the present disclosure. In another embodiment, the disorder is TBI, and at least one symptom of TBI is confusion, dizziness, disorientation, loss of coordination, memory loss, inability to form new memories, sleep disturbances, behavioral or mood changes, increased agitation, depression, convulsions, and / or seizures.

[0210] In another embodiment, administration of a compound of the present disclosure reduces the severity of at least one symptom of a disease, disorder, condition or syndrome, e.g., a disease, disorder, condition or syndrome provided in any one of Tables 1-17 (e.g., TBI) by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 70%, at least 90%, or 100% compared to baseline, such as compared to a subject with a disease, disorder, condition or syndrome, e.g., a disease, disorder, condition or syndrome provided in any one of Tables 1-17 (e.g., TBI) not administered leucine, acetyl-leucine, or a pharmaceutically acceptable salt thereof.

[0211] In another embodiment, administration of a Compound of the Disclosure is effective to improve the level of physical and / or mental activity in a subject in need thereof. In another embodiment, recovery is improved compared to recovery of a subject with a disease, disorder, condition, or syndrome, such as a disease, disorder, condition, or syndrome provided in any one of Tables 1-17 (e.g., TBI), who is not administered a Compound of the Disclosure.

[0212] In another embodiment, the administration of the compounds of the present disclosure effectively treats encephalitis. In another embodiment, encephalitis is infectious encephalitis. In another embodiment, encephalitis is autoimmune encephalitis. The symptoms of encephalitis include but are not limited to headache, mild flu-like symptoms (pain, fatigue, slight fever), sensitivity to light, stiff neck, sleepiness (sleepiness) or lethargy (lethargy), more irritability, epileptic seizures, changes in alertness, confusion or hallucinations, lack of energy, loss of appetite, unsteady gait, nausea and vomiting, personality changes and difficulty in talking and / or changes in speech. In some embodiments, the symptoms of encephalitis are abnormal movements (chorea, incoordination), atypical movement symptoms, such as ataxia or hemiparesis, or epileptic seizures.

[0213] In some embodiments, encephalitis is caused by an upper respiratory tract infection, a disease causing diarrhea, nausea and vomiting, measles, mumps, rubella, varicella, herpes simplex virus, West Nile virus, rabies, Lyme disease, tuberculosis, syphilis, an infection caused by a parasite (e.g., toxoplasmosis), or an autoimmune reaction of the subject. Therefore, in some embodiments, the present disclosure provides a method of preventing encephalitis in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of the present disclosure, wherein the subject suffers from an upper respiratory tract infection, a disease causing diarrhea, nausea and vomiting, measles, mumps, rubella, varicella, herpes simplex virus, West Nile virus, rabies, Lyme disease, tuberculosis, syphilis, an infection caused by a parasite (e.g., toxoplasmosis), or an autoimmune reaction.

[0214] In another embodiment, administration of the compounds of the present disclosure can be effective in treating sleep disorders associated with traumatic brain injury, such as difficulty falling asleep and / or staying asleep, fatigue, sleepiness, sleep apnea, narcolepsy, and / or hypersomnia, e.g., excessive daytime sleepiness.

[0215] In another embodiment, the administration of the compounds of the present disclosure is effective to treat metabolic abnormalities in the CNS. In another embodiment, the administration of the compounds of the present disclosure is effective to treat symptoms and / or diseases associated with metabolic abnormalities in the CNS. In another embodiment, the metabolic abnormality is an imbalance in glucose metabolism. In another embodiment, the metabolic abnormality is an imbalance in glucose metabolism in the cerebellum. In another embodiment, the administration of the compounds of the present disclosure increases the metabolism of glucose in, for example, the cerebellum, in the CNS.

[0216] The present disclosure also provides the following specific embodiments.

[0217] Embodiment I: Leucine, acetyl-leucine, or a pharmaceutically acceptable salt thereof, for use in a method of treating traumatic brain injury (TBI) in a subject in need thereof.

[0218] Embodiment II: Leucine, acetyl-leucine or a pharmaceutically acceptable salt thereof, for use in a method according to Embodiment I, wherein the method comprises administering to the subject a therapeutically effective amount of leucine, acetyl-leucine or a pharmaceutically acceptable salt thereof.

[0219] Embodiment III: Leucine, acetyl-leucine or a pharmaceutically acceptable salt thereof, for use in a method according to any one of embodiments I and II, wherein the leucine is DL-leucine or the acetyl-leucine is acetyl-DL-leucine.

[0220] Embodiment IV: Leucine, acetyl-leucine or a pharmaceutically acceptable salt thereof, for use in a method according to any one of Embodiments I and II, wherein the leucine or acetyl-leucine has an enantiomeric excess of the L-enantiomer or the D-enantiomer.

[0221] Embodiment V: Leucine, acetyl-leucine, or a pharmaceutically acceptable salt thereof, for use in a method according to any one of Embodiments I-IV, wherein the method comprises administering acetyl-leucine to a subject in need thereof in a therapeutically effective amount of about 1 g to about 15 g per day, about 1 g to about 10 g per day, about 1.5 g to about 7 g per day, about 4 g to about 6 g per day, or about 4 g to about 5 g per day.

[0222] Embodiment VI: A method of inhibiting or reducing one or more symptoms of traumatic brain injury (TBI) in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of leucine, acetyl-leucine, or a pharmaceutically acceptable salt thereof.

[0223] Embodiment VII: The method according to embodiment VI, wherein said leucine is DL-leucine or said acetyl-leucine is acetyl-DL-leucine.

[0224] Embodiment VIII: A method according to embodiment VI, wherein the leucine or acetyl-leucine has an enantiomeric excess of the L-enantiomer or the D-enantiomer.

[0225] Embodiment IX: The method according to any one of embodiments VI-VIII, wherein the therapeutically effective amount is about 1 g to about 15 g per day, about 1 g to about 10 g per day, about 1.5 g to about 7 g per day, about 4 g to about 6 g per day, or about 4 g to about 5 g per day.

[0226] The present disclosure also provides the following specific embodiments.

[0227] Embodiment 1. A compound of the present disclosure for use in treating a disease, disorder, condition or syndrome in a subject in need thereof, or for treating a symptom of a disease, disorder, condition or syndrome in a subject in need thereof, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 1.

[0228] Embodiment 2. A compound of the present disclosure for use as described in Embodiment 1, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 2.

[0229] Embodiment 3. A compound of the present disclosure for use as described in Embodiment 1, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 3.

[0230] Embodiment 4. A compound of the present disclosure for use as described in Embodiment 1, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 4.

[0231] Embodiment 5. A compound of the present disclosure for use as described in Embodiment 1, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 5.

[0232] Embodiment 6. A compound of the disclosure for use as described in Embodiment 1, wherein the disease, condition, disorder or syndrome is traumatic brain injury.

[0233] Embodiment 7. A compound of the present disclosure for use as described in Embodiment 1, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 6.

[0234] Embodiment 8. A compound of the present disclosure for use as described in Embodiment 1, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 7.

[0235] Embodiment 9. A compound of the present disclosure for use as described in Embodiment 1, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 8.

[0236] Embodiment 10. A compound of the present disclosure for use as described in Embodiment 1, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 9.

[0237] Embodiment 11. A compound of the present disclosure for use as described in Embodiment 1, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 10.

[0238] Embodiment 12. A compound of the present disclosure for use as described in Embodiment 1, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 11.

[0239] Embodiment 13. A compound of the present disclosure for use as described in Embodiment 1, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 12.

[0240] Embodiment 14. A compound of the disclosure for use as described in Embodiment 1, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 13.

[0241] Embodiment 15. A compound of the present disclosure for use as described in Embodiment 1, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 14.

[0242] Embodiment 16. A compound of the present disclosure for use as described in Embodiment 1, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 15.

[0243] Embodiment 17. A compound of the disclosure for use as described in Embodiment 1, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 16.

[0244] Embodiment 18. The compound of the present disclosure for use as described in any one of embodiments 1-17, wherein about 1 g to about 30 g of the compound of the present disclosure is administered to the subject per day.

[0245] Embodiment 19. The compound of the present disclosure for use as described in embodiment 18, wherein about 2 g to about 15 g of the compound of the present disclosure is administered to the subject per day.

[0246] Embodiment 20. The compound of the present disclosure for use as described in embodiment 19, wherein about 3 g to about 10 g of the compound of the present disclosure is administered to the subject per day.

[0247] Embodiment 21. The compound of the present disclosure for use as described in embodiment 20, wherein about 4 g to about 8 g of the compound of the present disclosure is administered to the subject per day.

[0248] Embodiment 22. The compound of the present disclosure for use as described in embodiment 21, wherein about 4 g to about 5 g of the compound of the present disclosure is administered to the subject per day.

[0249] Embodiment 23. The compound of the present disclosure for use as described in embodiment 22, wherein about 5 g of the compound of the present disclosure is administered to the subject per day.

[0250] Embodiment 24. A compound of the present disclosure for use as described in any one of embodiments 1-23, wherein a therapeutically effective amount of leucine or a pharmaceutically acceptable salt thereof is administered to the subject.

[0251] Embodiment 25. A compound of the present disclosure for use as described in Embodiment 24, wherein a therapeutically effective amount of leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the disease, condition, disorder or syndrome.

[0252] Embodiment 26. A compound of the present disclosure for use as described in Embodiment 24, wherein a therapeutically effective amount of leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the symptoms of the disease, condition, disorder or syndrome.

[0253] Embodiment 27. A compound of the present disclosure for use as described in any one of embodiments 1-23, wherein a therapeutically effective amount of acetyl-DL-leucine or a pharmaceutically acceptable salt thereof is administered to the subject.

[0254] Embodiment 28. A compound of the present disclosure for use as described in Embodiment 27, wherein a therapeutically effective amount of acetyl-DL-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the disease, condition, disorder or syndrome.

[0255] Embodiment 29. A compound of the present disclosure for use as described in Embodiment 27, wherein a therapeutically effective amount of acetyl-DL-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the symptoms of the disease, condition, disorder or syndrome.

[0256] Embodiment 30. A compound of the present disclosure for use as described in any one of embodiments 1-23, wherein a therapeutically effective amount of acetyl-L-leucine or a pharmaceutically acceptable salt thereof is administered to the subject.

[0257] Embodiment 31. A compound of the present disclosure for use as described in Embodiment 30, wherein a therapeutically effective amount of acetyl-L-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the disease, condition, disorder or syndrome.

[0258] Embodiment 32. A compound of the present disclosure for use as described in Embodiment 30, wherein a therapeutically effective amount of acetyl-L-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the symptoms of the disease, condition, disorder or syndrome.

[0259] Embodiment 32A. A compound of the present disclosure for use as described in any one of Embodiments 1-23, wherein a therapeutically effective amount of acetyl-D-leucine or a pharmaceutically acceptable salt thereof is administered to the subject.

[0260] Embodiment 32B. A compound of the present disclosure for use as described in Embodiment 32A, wherein a therapeutically effective amount of acetyl-D-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the disease, condition, disorder or syndrome.

[0261] Embodiment 32C. A compound of the present disclosure for use as described in Embodiment 32A, wherein a therapeutically effective amount of acetyl-D-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the symptoms of the disease, condition, disorder or syndrome.

[0262] Embodiment 33. Use of a compound of the present disclosure for the manufacture of a medicament for treating a disease, disorder, condition, or syndrome in a subject in need thereof, or for the manufacture of a medicament for treating a symptom of a disease, disorder, condition, or syndrome in a subject in need thereof, wherein the disease, disorder, condition, or syndrome is any one or more of the diseases, disorders, conditions, or syndromes provided in Table 1.

[0263] Embodiment 34. The use according to embodiment 33, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 2.

[0264] Embodiment 35. The use of embodiment 33, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 3.

[0265] Embodiment 36. The use of embodiment 33, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 4.

[0266] Embodiment 37. The use according to embodiment 33, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 5.

[0267] Embodiment 38. The use of embodiment 37, wherein the disease, condition, disorder or syndrome is traumatic brain injury.

[0268] Embodiment 39. The use of embodiment 33, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 6.

[0269] Embodiment 40. The use of embodiment 33, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 7.

[0270] Embodiment 41. The use of embodiment 33, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 8.

[0271] Embodiment 42. The use of embodiment 33, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 9.

[0272] Embodiment 43. The use of embodiment 33, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 10.

[0273] Embodiment 44. The use of embodiment 33, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 11.

[0274] Embodiment 45. The use of embodiment 33, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 12.

[0275] Embodiment 46. The use of embodiment 33, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 13.

[0276] Embodiment 47. The use of embodiment 33, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 14.

[0277] Embodiment 48. The use of embodiment 33, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 15.

[0278] Embodiment 49. The use of embodiment 33, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 16.

[0279] Embodiment 50. The use of any one of embodiments 33-49, wherein about 1 g to about 30 g of the compound of the disclosure is administered to the subject per day.

[0280] Embodiment 51. The use according to embodiment 50, wherein about 2 g to about 15 g of the compound of the present disclosure is administered to the subject per day.

[0281] Embodiment 52. The use according to embodiment 51, wherein about 3 g to about 10 g of the compound of the disclosure is administered to the subject per day.

[0282] Embodiment 53. The use according to embodiment 52, wherein about 4 g to about 8 g of the compound of the present disclosure is administered to the subject per day.

[0283] Embodiment 54. The use according to embodiment 53, wherein about 4 g to about 5 g of the compound of the present disclosure is administered to the subject per day.

[0284] Embodiment 55. The use according to embodiment 54, wherein about 5 g of the compound of the present disclosure is administered to the subject per day.

[0285] Embodiment 56. The use of any one of embodiments 33-55, wherein a therapeutically effective amount of leucine or a pharmaceutically acceptable salt thereof is administered to the subject.

[0286] Embodiment 57. The use according to embodiment 56, wherein a therapeutically effective amount of leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the disease, condition, disorder or syndrome.

[0287] Embodiment 58. The use according to embodiment 56, wherein a therapeutically effective amount of leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the symptoms of the disease, condition, disorder or syndrome.

[0288] Embodiment 59. The use of any one of embodiments 33-55, wherein a therapeutically effective amount of acetyl-DL-leucine or a pharmaceutically acceptable salt thereof is administered to the subject.

[0289] Embodiment 60. The use according to embodiment 59, wherein a therapeutically effective amount of acetyl-DL-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the disease, condition, disorder or syndrome.

[0290] Embodiment 61. The use according to embodiment 59, wherein a therapeutically effective amount of acetyl-DL-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the symptoms of the disease, condition, disorder or syndrome.

[0291] Embodiment 62. The use of any one of embodiments 33-55, wherein a therapeutically effective amount of acetyl-L-leucine or a pharmaceutically acceptable salt thereof is administered to the subject.

[0292] Embodiment 63. The use according to embodiment 62, wherein a therapeutically effective amount of acetyl-L-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the disease, condition, disorder or syndrome.

[0293] Embodiment 64. The use according to embodiment 62, wherein a therapeutically effective amount of acetyl-L-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the symptoms of the disease, condition, disorder or syndrome.

[0294] Embodiment 64A. The use of any one of Embodiments 33-55, wherein a therapeutically effective amount of acetyl-D-leucine or a pharmaceutically acceptable salt thereof is administered to the subject.

[0295] Embodiment 64B. The use according to embodiment 64A, wherein a therapeutically effective amount of acetyl-D-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the disease, condition, disorder or syndrome.

[0296] Embodiment 64C. The use of embodiment 64A, wherein a therapeutically effective amount of acetyl-D-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the symptoms of the disease, condition, disorder or syndrome.

[0297] Embodiment 65. A pharmaceutical composition for treating a disease, disorder, condition or syndrome in a subject in need thereof, or for treating a symptom of a disease, disorder, condition or syndrome in a subject in need thereof, the pharmaceutical composition comprising a compound of the present disclosure and a pharmaceutically acceptable carrier, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 1.

[0298] Embodiment 66. A pharmaceutical composition for use as described in Embodiment 65, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 2.

[0299] Embodiment 67. A pharmaceutical composition for use as described in Embodiment 65, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 3.

[0300] Embodiment 68. A pharmaceutical composition for use as described in Embodiment 65, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 4.

[0301] Embodiment 69. A pharmaceutical composition for use as described in Embodiment 65, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 5.

[0302] Embodiment 70. A pharmaceutical composition for use according to embodiment 65, wherein the disease, condition, disorder or syndrome is traumatic brain injury.

[0303] Embodiment 71. A pharmaceutical composition for use as described in Embodiment 65, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 6.

[0304] Embodiment 72. A pharmaceutical composition for use as described in Embodiment 65, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 7.

[0305] Embodiment 73. A pharmaceutical composition for use as described in Embodiment 65, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 8.

[0306] Embodiment 74. A pharmaceutical composition for use as described in Embodiment 65, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 9.

[0307] Embodiment 75. A pharmaceutical composition for use as described in Embodiment 65, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 10.

[0308] Embodiment 76. A pharmaceutical composition for use as described in Embodiment 65, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 11.

[0309] Embodiment 77. A pharmaceutical composition for use as described in Embodiment 65, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 12.

[0310] Embodiment 78. A pharmaceutical composition for use as described in Embodiment 65, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 13.

[0311] Embodiment 79. A pharmaceutical composition for use as described in Embodiment 65, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 14.

[0312] Embodiment 80. A pharmaceutical composition for use as described in Embodiment 65, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 15.

[0313] Embodiment 81. A pharmaceutical composition for use as described in Embodiment 65, wherein the disease, disorder, condition or syndrome is any one or more of the diseases, disorders, conditions or syndromes provided in Table 16.

[0314] Embodiment 82. A pharmaceutical composition for use as described in any one of embodiments 65-81, wherein about 1 g to about 30 g of a compound of the disclosure is administered to the subject per day.

[0315] Embodiment 83. A pharmaceutical composition for use as described in Embodiment 82, wherein about 2 g to about 15 g of a compound of the disclosure is administered to the subject per day.

[0316] Embodiment 84. A pharmaceutical composition for use as described in Embodiment 83, wherein about 3 g to about 10 g of a compound of the disclosure is administered to the subject per day.

[0317] Embodiment 85. A pharmaceutical composition for use as described in Embodiment 84, wherein about 4 g to about 8 g of a compound of the disclosure is administered to the subject per day.

[0318] Embodiment 86. A pharmaceutical composition for use as described in Embodiment 85, wherein about 4 g to about 5 g of a compound of the disclosure is administered to the subject per day.

[0319] Embodiment 87. A pharmaceutical composition for use as described in embodiment 86, wherein about 5 g of a compound of the present disclosure is administered to the subject per day.

[0320] Embodiment 88. A pharmaceutical composition for use as described in any one of embodiments 65-87, wherein a therapeutically effective amount of leucine or a pharmaceutically acceptable salt thereof is administered to the subject.

[0321] Embodiment 89. A pharmaceutical composition for use as described in embodiment 88, wherein a therapeutically effective amount of leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the disease, condition, disorder or syndrome.

[0322] Embodiment 90. A pharmaceutical composition for use as described in embodiment 88, wherein a therapeutically effective amount of leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the symptoms of the disease, condition, disorder or syndrome.

[0323] Embodiment 91. A pharmaceutical composition for use as described in any one of embodiments 65-87, wherein a therapeutically effective amount of acetyl-DL-leucine or a pharmaceutically acceptable salt thereof is administered to the subject.

[0324] Embodiment 92. A pharmaceutical composition for use as described in embodiment 91, wherein a therapeutically effective amount of acetyl-DL-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the disease, condition, disorder or syndrome.

[0325] Embodiment 93. A pharmaceutical composition for use as described in Embodiment 91, wherein a therapeutically effective amount of acetyl-DL-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the symptoms of the disease, condition, disorder or syndrome.

[0326] Embodiment 94. A pharmaceutical composition for use as described in any one of embodiments 65-87, wherein a therapeutically effective amount of acetyl-L-leucine or a pharmaceutically acceptable salt thereof is administered to the subject.

[0327] Embodiment 95. A pharmaceutical composition for use as described in Embodiment 94, wherein a therapeutically effective amount of acetyl-L-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the disease, condition, disorder or syndrome.

[0328] Embodiment 96. A pharmaceutical composition for use as described in Embodiment 94, wherein a therapeutically effective amount of acetyl-L-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the symptoms of the disease, condition, disorder or syndrome.

[0329] Embodiment 96A. A pharmaceutical composition for use as described in any one of embodiments 65-87, wherein a therapeutically effective amount of acetyl-D-leucine or a pharmaceutically acceptable salt thereof is administered to the subject.

[0330] Embodiment 96B. A pharmaceutical composition for use as described in Embodiment 96A, wherein a therapeutically effective amount of acetyl-D-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the disease, condition, disorder or syndrome.

[0331] Embodiment 96C. A pharmaceutical composition for use as described in Embodiment 96A, wherein a therapeutically effective amount of acetyl-D-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the symptoms of the disease, condition, disorder or syndrome.

[0332] Example

[0333] Example 1

[0334] Acetyl-DL-leucine was administered to a male patient with traumatic brain injury at a dose of 3 g per day for the first week and then 5 g per day. The patient's therapist and nurse reported that the patient exhibited more active behavior after treatment with acetyl-DL-leucine. The medication was later discontinued and the patient's activity level returned to the original level before treatment with acetyl-DL-leucine.

[0335] Example 2

[0336] Efficacy of N-acetyl-L-leucine in the CCI mouse model of TBI

[0337] Controlled cortical impact model

[0338] Controlled cortical impact (CCI) induced TBI was performed in male C57BL6 / J mice (20-25 g). Briefly, after a 10-mm midline incision on the skull, the skin and fascia were retracted and a 4 mm craniotomy was performed in the central portion of the left parietal bone of the mouse under surgical anesthesia (2%-3% isoflurane vaporized in a gas mixture containing 70% N2O and 30% O2). Moderate injury was induced in the exposed brain by a custom microprocessor-controlled and compressed air-driven 3.5 mm diameter tip pneumatic impactor with an impact velocity of 6 m / s and a deformation depth of 2 mm.

[0339] NAL Treatment

[0340] N-acetyl-L-leucine ("NAL" or "NALL") was dissolved in ethanol to prepare a 50 mg / ml solution, which was then diluted in water to give a 10 mg / ml solution. Four days after CCI-induced TBI, mice were orally administered approximately 0.25 ml of NAL solution (10 mg / ml) daily by oral gavage at a dose of 10 mg / kg (2.5 mg NAL / 25 g mouse). Mice were also fed NAL at 0.5 mg / kg of food powder mixed with dietary gel for up to 7 days after injury. See Figure 1 No significant differences in food intake were observed between sham-operated mice and TBI mice fed with NALL (data not shown). Although a slight decrease in body weight was detected in vehicle-treated TBI mice, no significant changes in body weight were observed in NALL-treated TBI mice (data not shown).

[0341] Cell Death After TBI

[0342] The levels of α-fodrin cleavage products in the cortex of NALL-treated, vehicle-treated sham-operated, and TBI mice were determined by Western blot analysis and using the TUNEL assay to assess whether NALL treatment reduces cortical cell death after TBI.

[0343] Western blot analysis

[0344] About 5 mm of tissue from the ipsilateral cortex around the injury site of TBI mice or the same volume of corresponding tissue around the same cortical area of ​​sham-operated mice was removed and processed. Tissue lysates were resolved on 4%-20% SDS-PAGE gels (Bio-Rad, 5671095) and transferred to PVDF membranes (Millipore, IPVH00010) using semi-dry transfer (Bio-Rad), blocked with 5% skim milk in tris-buffered saline containing 0.05% tween 20 (TBST), probed with primary antibodies in 1% BSA in TBST at 4°C overnight, and incubated with HRP-conjugated secondary antibodies (KPL, 474-1506, 474-1806, 14-16-06 and 14-13-06) in blocking solution at room temperature for 1 hour. Protein bands were detected using a chemiluminescent kit (Pierce, 34076) and visualized using the Chemi-doc system (Bio-Rad). Band intensities were analyzed using Image Lab software (Bio-Rad) and normalized to a loading control (β-actin). Primary antibodies: LC3 (1:1000; Novus, NB100-2220), p62 / SQSTM1 (1:1000; BD Bioscience, 610832), β-actin / ACTB (1:10,000; A1978), and fodrin / spectrin (1:5000; Enzo Life Science International, BML-FG6090).

[0345] TUNEL assay

[0346] From the sham-operated mice and TBI mice brains that were fixed with 4% paraformaldehyde (PFA, pH 7.4) and protected in 30% sucrose or NAL fed frozen sections (20 μm). According to the manufacturer's protocol, ApopTag in situ apoptosis detection kit (Millipore, S7165) was used to perform TUNEL assay on frozen brain sections. Fluorescence Nikon Ti-E inverted microscope was used to obtain images after TUNEL assay at 20X (CFIPlan APO VC 20X NA 0.75WD 1mm).

[0347] TBI induces calpain- and caspase-mediated cleavage of α-fodrin, generating 145-150 kDa and 150-120 kDa fragments, respectively. Lower levels of the 145-150 kDa fragment of α-fodrin were observed in the cortex of mice fed NALL compared to the cortex of vehicle-fed mice after TBI, suggesting a protective benefit of NALL in preventing cortical cell death. Fig.28 and 29 Lower levels of TUNEL-positive cells were also detected in brain sections from injured mice fed NALL compared to TBI mice fed vehicle, suggesting a protective role for NALL in attenuating cell death after TBI. Fig.30 , 31 and 32.

[0348] Autophagic flux after TBI

[0349] Autophagy is a lysosome-dependent cellular degradation process that is disrupted due to lysosomal damage after TBI. See, e.g., Sarkar et al., Autophagy 10:2208-2222 (2014); Sarkar et al., Autophagy, 1-20 (2019). We evaluated whether the attenuation of cell death after NALL treatment in TBI mice was mediated by restoration of autophagic flux. The results are presented in Figure 11-13 and 33-35.

[0350] LC3-II and p62 / SQSTM1 levels were determined by Western blot analysis in cortical tissue lysates prepared from vehicle- and NAL-fed naive and TBI mice. LC3-II levels were reduced in the brains of NALL-fed mice compared to vehicle-treated mice as early as day 1 after TBI. Fig.33 and 34 We also observed a decrease in p62 / SQSTM1 levels in the cortex of mice fed NALL compared with vehicle-treated mice after TBI. Fig.33 and 35 These results suggest that NALL improves autophagic flux and thereby restores neuroprotection in the mouse cortex after TBI.

[0351] Inflammatory cytokines in the brain of TBI mice

[0352] The expression levels of M1 proinflammatory markers Nos2, Nlrp3, IL-1β, TNF, IFNβ, and Nox2 in the perilesional areas of the cortex of TBI mice were determined by real-time RT-PCR. The expression levels of M2 markers Socs3, YM-1, IL4ra, and Arg-1 in cortical tissue were also determined. The results are presented in Figure 2-10 and Figure 36-45 middle.

[0353] Real-time PCR

[0354] Total RNA isolated from the ipsilateral cortex using the miRNeasy Mini Kit (Qiagen, Catalog No. 217004) was converted to cDNA using the High Capacity RNA to cDNA Kit (Applied Biosystem, Catalog No. 4387406) according to the manufacturer's instructions. Universal Master Mix II (Applied Biosystems, catalog number 4440040) was used for quantitative real-time PCR amplification as described previously, using Gene expression assays (Applied Biosystems) were performed using the following mouse genes: Gapdh (Mm99999915_g1), NLRP3 (Mm00840904_m1), NOX2 (Mm01287743_m1), iNOS (Mm00440502_m1), TNFα (Mm00443258_m1), IFNβ1 (Mm00439 552_s1), IL-10 (Mm01288386_m1), IL1b (Mm00434228_m1), Arg1 (Mm00475988_m1), SOCS3 (Mm00545913_s1), YM-1 (Mm00657889_mH) and IL-4ra (Mm01275139_m1) (Applied Biosystems). The reaction was amplified and quantified using the 7900HT fast real-time PCR system and corresponding software (Applied Biosystems). Relative gene expression normalized to Gapdh was calculated based on the comparative Ct method.

[0355] Elevated levels of all M1-type proinflammatory markers were observed, starting at day 1 and peaking at day 3 after cortical injury in all TBI mice, independent of treatment. Figure 36-41 However, it was observed that the mRNA levels of IFNβ and Nox2 were decreased in the cortex of NAL-fed mice compared with the cortex of vehicle-fed TBI mice.

[0356] Similar to the M1-type markers, higher expression of the M2-type markers Socs3, YM-1, IL4ra, and Arg-1 was observed in the cortical tissue of all TBI mice compared to sham-operated animals. Figure 42-45Among these markers, NAL treatment reduced the level of Arg-1 in the cortex of injured mice compared with vehicle-fed TBI mice. The levels of Socs3, YM-1, and IL-4ra remained unchanged in the cortex of NAL-fed mice compared with sham-operated mice.

[0357] These results demonstrate that NAL reduced the expression of several inflammatory markers, thereby indicating an overall reduction in neuroinflammation after TBI in mice.

[0358] Statistics

[0359] All data are presented as mean ± standard error of the mean (SEM). One-way ANOVA was performed followed by appropriate post hoc tests (Tukey's) as specified in the figure legends. For data with only two groups, a 2-tailed Student's t-test with equal variance was used. p values ​​≤ 0.05 were considered statistically significant.

[0360] Example 3

[0361] EAL flux in NPC mice

[0362] Accumulation of amyloid β deposition and autophagic vacuoles is a pathological feature of Alzheimer's disease (AD). Dysregulation of the endosome-autophagy-lysosome (EAL) pathway, which impairs the processing of amyloid precursor protein, is one of the earliest changes in AD. Xu and Ren, Annu Rev Physiol. 77: 57-80 (2015). Lysosomal storage diseases (such as NPC) also show autophagy dysfunction, and their pathological changes are reminiscent of the features seen in AD, such as dystrophic axons, ectopic dendrites, neurofibrillary tangles, and amyloid aggregation. Boland et al., The Journal of Biological Chemistry 285: 37415–37426 (2010).

[0363] The efficacy of ADLL, ALL, ADL, DLL, LL, DL, and LLE in alleviating impaired EAL flux in NPC1 mice was investigated. Acetyl DL-leucine (ADLL) and L-leucine ethyl ester (LLE) effectively restored autophagic flux, thereby reducing LC3-II levels ( Fig.14 However, the reduction in LC3-II levels caused by ADLL and LLE was not accompanied by a decrease in APP-CTF, especially CTF-6 and -7 ( Fig.15Without wishing to be bound by any particular theory, it is possible that ADL and LLE reduce autophagic vacuoles (AV) accumulation by inhibiting autophagosome formation, possibly through activation of mTORC1. Although they were able to reduce AV accumulation, their inability to reduce elevated APP-CTF levels suggests that ADL and LLE do not increase lysosomal digestion capacity per se. Surprisingly, L-leucine did reduce the accumulation of CTF-6 in NPC1 mice ( Fig.16 ). In addition, acetyl L-leucine (ALL), acetyl D-leucine (ADL), and L-leucine (LL) decreased CTF-7 expression, suggesting some improvement in lysosomal proteolysis ( Fig.17 The positive effects of acetyl D-leucine, acetyl L-leucine, and D-leucine on CTF-7 clearance in NPC1 mice suggest that they promote endosome-lysosome fusion without affecting autophagosome-lysosome clearance.

[0364] Example 4

[0365] Lysosomal fusion in primary cortical neurons

[0366] Neurons have a heterogeneous distribution of organelles belonging to the EAL pathway, ranging from cell bodies to distal neurites. These include autophagosomes, endosomes and amphisomes (endosomes fused with autophagosomes). U18666A is an agent that induces NPC1-like phenotypes in cells by inhibiting NPC1 protein and preventing endosome-lysosome fusion. The elimination of autophagosome-lysosome fusion and endosome-lysosome fusion mediated by U18666A leads to an increase in LC3-II and APP-CTF-1 to -5, while preventing the generation of CTF-6 and -7. Boland et al., The Journal of Biological Chemistry 285:37415–37426 (2010).

[0367] The efficacy of ADLL, ALL, ADL, DLL, LL, DL, and LLE in alleviating U18666A-mediated impairment of autophagosome-lysosome fusion and endosome-lysosome fusion was investigated. In this cell-based EAL flux impairment assay, basal levels of APP CTF-6 and -7 reappeared, indicating improved endosome-lysosome fusion.

[0368] Rat primary cortical neurons (DIV14) treated with U18666A (2 μg / mL, 24 hours) completely lacked CTF-6 and 7 ( Fig.18 U18666A treatment for 24 hours did not result in an increase in LC3-II accumulation ( Fig.18 ).

[0369] Neurons were co-treated with U18666A (2 μg / mL) and ADLL, ALL, ADL, DLL, LL, DL, and LLE (1 mM, 24 hours). All co-treated neuronal cultures showed signs of improved endosome-lysosome fusion, as evidenced by the reappearance of basal amounts of CTF-6 and -7 in these cells ( Fig.18 ). ADLL, ALL, and ADL reduced LC3-II levels below baseline ( Fig.18 ). This suggests that the three compounds enhance autophagosome-lysosome fusion and endosome-lysosome fusion in primary cortical neurons.

[0370] Example 5

[0371] Truncated forms of tau in neuron-glia co-cultures

[0372] The efficacy of ADLL, ALL, ADL, DLL, LL, DL, and LLE in reducing truncated tau levels in neuronal-glial (NG) cultures was investigated. The results of this study are provided in Figure 20-24 One objective of this study was to determine whether ADLL, ALL, ADL, DLL, LL, DL, and LLE confer intrinsic neuroprotection by maintaining neuronal function under conditions of cellular overgrowth, or whether they act indirectly through nonautonomous regulation of glial cells. Regardless of whether the compounds act directly or indirectly on neurons, preventing the production of truncated tau has a beneficial effect on neuronal integrity.

[0373] Cultures were treated with 1 mM ADLL, ALL, ADL, DLL, LL, DL, or LLE for five days (DIV 9-14) and subsequently assessed for a number of neuron-specific markers that provide a readout of neuronal health: βIII-tubulin, full-length tau, truncated tau (antibody 5E2), and synaptophysin. ADLL, ALL, ADL, DLL, LL, DL, and LLE were well tolerated in both NE and NG cultures, as demonstrated by consistently similar levels of synaptophysin and βIII-tubulin detected in untreated (control) and drug-treated samples ( Fig. 20 , 23 and 24). ADLL, ALL, ADL, DLL, LL, DL, and LLE did not reduce the basal level of truncated tau in NG cultures at DIV14 ( Fig. 20 and Fig. 22 ). These findings suggest that ADLL, ALL, ADL, DLL, LL, DL, and LLE are particularly useful for treating neurodegenerative disorders involving accumulation of substrates in the EAL network.

[0374] Example 6

[0375] Lysosomal volume measured by flow cytometry

[0376] Wild-type and NPC1-deficient Chinese hamster ovary (CHO) cells were grown in T75 culture flasks and treated with N-acetyl-L-leucine (ALL), L-leucine ethyl ester hydrochloride (LEE) or L-leucine (1 or 5 mM, purchased from Sigma-Aldrich) for 7 days and then extracted. The cells were trypsinized, centrifuged (270 x g, 5 minutes), washed twice with 1x PBS, centrifuged again and stained with 100 nM LysoTracker-green DND-26 (Invitrogen) in PBS for 20 minutes at room temperature. After incubation, the cells were centrifuged (800 x g, 5 minutes), resuspended in 0.5 ml FACS buffer (0.1% BSA, 0.02 M NaN3 in 1×PBS) and stored on ice for flow cytometry analysis (BD Biosciences FACSCanto II or Accuri C6 Plus). The hemacytometer was calibrated before the assay using Cytometer Setup and Tracking beads (BD) and compensated using Lysotracker or propidium iodide-stained cells using BD FACSDiva software (BD) or BD Accuri C6 Plus software (BD). 10,000 recorded cells were analyzed to obtain the median value for further statistical analysis. Results are presented in Figure 25-27 Shown in.

[0377] Example 7

[0378] Purkinje cell survival

[0379] Npc1- / - mice exhibit progressive neurodegeneration, with cerebellar Purkinje cell loss progressing from the anterior to the posterior lobes, accompanied by microglial activation ( Fig.46 ). ADLL treatment increased Purkinje cell survival at 59 days of age compared with untreated Npc1- / - littermates: there were 133% more Purkinje cells in lobules I and II (p=0.0027), and 402% more Purkinje cells in lobule III (p=0.0108). Other treatments did not increase Purkinje cell survival in any cerebellar lobule (Npc1- / -ALL lobule I-II p=0.107, lobule III p=0.157, lobule IV-V p=0.533; Npc1- / -ADL lobule I-II p=0.60, lobule III p=0.11, lobule IV-V p=0.766 compared with Npc1- / -UT) ( Fig.46 and47 In addition, ALL treatment reduced (by 20%, p=0.0177) the frequency of CD68-positive activated microglia. Other treatments had no effect (p=0.1353 for Npc1- / -ADLL and p=0.0553 for Npc1- / -ADL compared to Npc1- / -UT) ( Fig.46 and 48 ADL did not mediate any long-term neuroprotective effects. The combination of increased neuronal survival and reduced brain inflammatory environment may improve brain function, quality of life and slow disease progression in human subjects.

[0380] Example 8

[0381] Controlled cortical impact model

[0382] Mice were subjected to either moderate controlled cortical impact TBI or sham surgery. NAL mice were treated with oral NAL starting after initial recovery after surgery (by oral gavage, once daily for 4 days) and continued for the duration of the experiment (food ad libitum). Vehicle mice received equivalent administrations of oral gavage vehicle followed by standard chow. Groups: sham + vehicle = 5 mice; sham + NAL = 4 mice; TBI + vehicle = 10 mice; and TBA + NAL = 9 mice. Functional outcomes were assessed up to 28 days after surgery in the beam walking (motor function), Y-maze (spatial memory), and novel object recognition (NOR, non-spatial memory) assessments. Results are presented in Fig.49 , Fig.50 and Fig.51 Available in.

[0383] It should be understood that the foregoing embodiments and examples are not intended to limit the scope of the present disclosure in any respect, and that the claims set forth herein are intended to cover all embodiments and examples, whether or not explicitly set forth herein.

[0384] All patents and publications cited herein are fully incorporated by reference in their entirety.

Claims

1. Use of acetyl-L-leucine or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating a disease, disorder or condition of a subject in need thereof or treating a symptom of the disease, disorder or condition of a subject in need thereof, wherein: The disease, disorder or condition is medial aortic amyloidosis, apathy, attention deficit disorder (ADD), attention deficit hyperactivity disorder (ADHD), autism, autism spectrum disorder (ASD), bipolar disorder, brain inflammation, brain injury, catatonia, cerebral amyloid angiopathy, cerebral hemorrhage with amyloidosis, cerebral palsy, cerebrovascular disease, chorea in children and adults, complications of cerebral hemorrhage, post-anoxia complications, concussion, conus and cauda equina syndrome, dementia, depression, type 1 diabetes, diffuse Lewy body disease, dopamine-responsive dystonia, drop foot, dyslexia, Emory Dreifuss muscular dystrophy, encephalitis (infectious or autoimmune), epilepsy, frontal lobe syndrome, frontotemporal lobar degeneration, generalized anxiety disorder, hydrocephalus, hyperalgesia or sensory disturbances, hypercalcemia, hyperkinesia, hypoglycemia, hypothyroidism, depression, malnutrition, mania, mild cognitive impairment, mitochondrial disorder, mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes (MELAS), moyamoya disease, multi-infarct dementia, neurotrauma, neurodegeneration with brain iron accumulation, neuroinflammation, neuropsychiatric disorders, pain, panic disorder, pantothenate kinase-associated neurodegeneration, Pick's disease, post-traumatic stress disorder, progressive bulbar palsy, progressive muscular atrophy, schizophrenia, social phobia, stroke, tardive dyskinesia, Tourette syndrome, transverse myelitis, traumatic brain injury, vascular amyloidosis, vascular dementia, or vasculitis.

2. The use according to claim 1, wherein: The disease, disorder or condition is Tourette syndrome.

3. The use according to claim 1, wherein: The disease, disorder or condition is attention deficit hyperactivity disorder (ADHD).

4. The use according to any one of claims 1 to 3, wherein: 1 g to 30 g of acetyl-L-leucine or a pharmaceutically acceptable salt thereof is administered to the subject daily.

5. The use according to claim 4, wherein: 2 g to 15 g of acetyl-L-leucine or a pharmaceutically acceptable salt thereof is administered to the subject daily.

6. The use according to claim 5, wherein: 3 g to 10 g of acetyl-L-leucine or a pharmaceutically acceptable salt thereof is administered to the subject daily.

7. The use according to claim 6, wherein: 4 g to 8 g of acetyl-L-leucine or a pharmaceutically acceptable salt thereof is administered to the subject daily.

8. The use according to claim 7, wherein: 4 g to 5 g of acetyl-L-leucine or a pharmaceutically acceptable salt thereof is administered to the subject daily.

9. The use according to claim 8, wherein: 5 g of acetyl-L-leucine or a pharmaceutically acceptable salt thereof is administered to the subject daily.

10. The use according to any one of claims 1 to 9, wherein: A therapeutically effective amount of acetyl-L-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the disease, disorder or condition.

11. The use according to any one of claims 1 to 9, wherein: A therapeutically effective amount of acetyl-L-leucine or a pharmaceutically acceptable salt thereof is administered to the subject to treat the symptoms of the disease, disorder or condition.

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