Pharmaceutical composition and use for lysosomal storage diseases

Abstract

The present invention relates to pharmaceutical compositions and uses against lysosomal storage disorders. The present disclosure provides treatment of lysosomal storage disease (LSD) comprising administering acetyl-leucine or a pharmaceutically acceptable salt thereof.

Description

[0001] This application is a divisional application. The original application has the International Application No. PCT / IB2017 / 054928, the filing date of 11 August 2017, the Chinese National Phase Application No. 201780062740.X, and the invention title of "Pharmaceutical compositions and uses for lysosomal storage disorders".

[0002] This application claims priority from UK 1613828.1 filed on 11 August 2016, UK 1702552.9 filed on 16 February 2017, UK 1705762.1 filed on 10 April 2017, and UK 1706854.5 filed on 28 April 2017; all of which are incorporated herein by reference in their entirety.

[0003] Lysosomal storage disorders (LSDs) are a group of inherited metabolic diseases caused by defects in lysosomal homeostasis. To date, LSDs include over 70 diseases with a clinical frequency of 1 :5000 live births. These diseases can be divided into two broad categories: primary storage disorders caused by a direct deficiency in a degradation pathway (typically a lysosomal enzyme deficiency), and secondary storage disorders caused by a downstream lysosomal protein malfunction or a process that affects the lysosome (e.g. a defect in a transport pathway).

[0004] The pathology of LSDs affects many systems of the body, but most commonly the nervous system. Progressive neurodegeneration leading to physical disability and mental deterioration is a common symptom. The diseases are typically severe, progressive and unrelenting. They tend to present in the first years of life, and severe progression leads to frequent hospitalisation. If left untreated, patients often die in their teens. Adult-onset patients have also been described.

[0005] Current treatment options for LSDs are limited. Few treatments exist, and if they do, many treatment options can only improve quality of life. For example, certain LSDs respond to bone marrow transplantation or enzyme replacement therapy. In addition, certain benefits have been reported in a clinical trial using substrate reduction therapy (SRT) with the glycosphingolipid (GSL) biosynthesis inhibitor: the iminosugar drug Miglustat (Patterson, 2006). However, there is currently no universal, non-specific treatment that benefits all LSDs. There is therefore a need to develop improved treatment options for LSDs.

[0006] The present disclosure addresses this need and describes acetyl-leucine for use in treating an LSD or one or more symptoms of an LSD in a subject in need thereof.

[0007] In one embodiment, acetyl-leucine or a pharmaceutically acceptable salt thereof is disclosed for use in a method of treating LSD or one or more symptoms associated with LSD in a subject in need thereof, wherein the LSD is not Niemann Pick Type C disease.

[0008] In one embodiment of the disclosure, acetyl-leucine or a pharmaceutically acceptable salt thereof is disclosed for use in a method of treating LSD in a subject in need thereof, wherein the subject is asymptomatic.

[0009] In another embodiment, acetyl-leucine or a pharmaceutically acceptable salt thereof is disclosed for use in a method of delaying the onset of LSD or one or more symptoms of LSD that would otherwise be expected to occur according to typical disease progression.

[0010] In yet another embodiment, the disclosure includes acetyl-leucine or a pharmaceutically acceptable salt thereof for use in a method of treating LSD or one or more symptoms associated with LSD in a subject in need thereof, wherein the method comprises administering to the subject in need thereof a therapeutically effective amount of acetyl-leucine for a duration selected from the group consisting of at least about 3 months, at least about 6 months, at least about 1 year, at least about 2 years, and at least about 5 years.

[0011] In one embodiment, the disclosure describes acetyl-leucine or a pharmaceutically acceptable salt thereof for use in a method of delaying the progression of LSD or one or more symptoms associated with LSD over time as compared to typical disease progression, wherein the method comprises administering to a subject in need thereof a therapeutically effective amount of acetyl-leucine for a duration selected from the group consisting of at least about 3 months, at least about 6 months, at least about 1 year, at least about 2 years, and at least about 5 years.

[0012] In yet another embodiment, acetyl-leucine or a pharmaceutically acceptable salt thereof is disclosed for use in a method of reversing the progression of LSD or one or more symptoms associated with LSD over time, wherein the method comprises administering to a subject in need thereof a therapeutically effective amount of acetyl-leucine for a duration selected from the group consisting of at least about 3 months, at least about 6 months, at least about 1 year, at least about 2 years, and at least about 5 years.

[0013] In another embodiment, acetyl-leucine or a pharmaceutically acceptable salt thereof is disclosed for use in a method of improving a biochemical marker of LSD over time in a subject in need thereof, wherein the method comprises administering to the subject in need thereof a therapeutically effective amount of acetyl-leucine for a duration selected from the group consisting of at least about 3 months, at least about 6 months, at least about 1 year, at least about 2 years, and at least about 5 years.

[0014] In another embodiment, the present disclosure includes acetyl-leucine, or a pharmaceutically acceptable salt thereof, for use in a method of reducing the severity of LSD, or reducing the severity of or eliminating one or more existing symptoms associated with LSD, in a subject in need thereof, wherein the LSD is not Niemann-Pick Type C disease.

[0015] In yet another embodiment, the present disclosure includes acetyl-leucine, or a pharmaceutically acceptable salt thereof, for use in a method of increasing neuroprotection in a subject having, suspected of having, or at risk of having LSD, wherein the method comprises administering to the subject a therapeutically effective amount of acetyl-leucine for a duration selected from the group consisting of at least about 3 months, at least about 6 months, at least about 1 year, at least about 2 years, and at least about 5 years.

[0016] Additional embodiments of the present disclosure include acetyl-leucine or a pharmaceutically acceptable salt thereof for use in a method of delaying progression of a lysosomal storage disorder (LSD) in a subject. Acetyl-leucine or a pharmaceutically acceptable salt thereof for use in a method of providing neuroprotection to a subject suffering from a LSD. The acetyl-leucine is in the racemic form, enantiomeric excess of the L enantiomer, or enantiomeric excess of the D enantiomer. The method further includes administering acetyl-leucine at a dose of 1.5 grams to 10 grams per day. Further, the method further includes administering acetyl-leucine for a treatment duration of more than two weeks. The method can further include administering acetyl-leucine or a pharmaceutically acceptable salt thereof prior to the onset of symptoms of a LSD. The method can further include administering another therapy or agent for the prevention or treatment of a LSD. A further embodiment of the present disclosure is a kit for delaying progression of a LSD in a subject, the kit comprising: a device for diagnosing or predicting a LSD, and acetyl-leucine or a pharmaceutically acceptable salt thereof. The kit comprises: a device for diagnosing or predicting a LSD, and acetyl-leucine or a pharmaceutically acceptable salt thereof. Yet another embodiment of the present disclosure is the use of acetyl-leucine or a pharmaceutically acceptable salt thereof as a neuroprotective agent in a subject suffering from a LSD. In further embodiments of the method, kit, or use, the LSD is Niemann-Pick disease type C (NPC1 and / or NPC2 deficiency), Smith-Lemli-Opitz Syndrome (SLOS), an inborn error of cholesterol synthesis, Tangier disease, Pelizaeus-Merzbacher disease, neuronal ceroid lipofuscinosis, primary glycosphingolipidosis, Farber disease, or multiple sulfatase deficiency. Further, in another embodiment of the method, kit, or use, the primary glycosphingolipidosis is Gaucher disease, Fabry disease, GM1 gangliosidosis, GM2 gangliosidosis, Krabbe disease, or metachromatic leukodystrophy (MLD).In further embodiments of the method, kit or use, the LSD is NPC, Tay-Sachs disease, Sandhoff disease, GM1 gangliosidosis, Fabry disease, neurodegenerative mucopolysaccharidosis, MPS I, MPS IH, MPS IS, MPS II, MPS III, MPS IIIA, MPS IIIB, MPS IIIC, MPS HID, MPS, IV, MPS IV A, MPS IV B, MPS VI, MPS VII, MPS IX, a disease with secondary lysosomal involvement, SLOS, or Danon disease. In another embodiment of the method, kit or use, the LSD is Niemann Pick disease, Niemann Pick disease type C, Niemann Pick disease type A, Sandhoff disease, Tay-Sachs disease, or Mucolipidosis II.

[0017] These and other embodiments and features of the present disclosure will be apparent from the description and claims that follow. BRIEF DESCRIPTION OF DRAWINGS

[0018] Figure 1 shows photographs of treated (left panel) Figure 1A ) and untreated (right panel) Figure 1B ) 9-week-old mice. Npc1 - / -

[0019] Figure 2A and 2B show that, with and without acetyl-DL-leucine treatment from weaning, Npc1 - / - weight data for mice compared to wild-type Npc1 + / + mice.

[0020] Npc1 show that, with and without acetyl-DL-leucine treatment from weaning, Figures 3A-3G - / - gait analysis data for mice compared to wild-type Npc1 + / + mice. For example, forepaw support, stride frequency, and stride sequence data are shown in Npc1 . Npc1 3E show forepaw (FP) data (average number of stands and stride cycle in inset D; duty cycle in inset E). Figures 3A-3C 3G ​​​Hindpaw (HP) data (average number of stands and step cycle in inset F; duty cycle in inset G) are shown.

[0021] Figure 3D It is shown that treatment with acetyl-DL-leucine from weaning onwards, Figure 3F - / - Motor function analysis data comparing mice to wild-type ( Figures 4A-4H + / + ) mice. Center-feeding, activity, rearing, and fore-to-rear (FR) counts are shown in Npc1 Activity time, movement time, rearing time, and total manual rearing counts are shown in Npc1

[0022] Figures 4A-4D It is shown that treatment with acetyl-DL-leucine (0.1 g / kg from 3 weeks of age) is associated with a small but statistically significant increase in lifespan of Npc1- / - mice.

[0023] Figures 4E-4H and 6B It is shown that lysosomal volume is reduced in non-neuronal NPC cells after treatment with acetyl-DL-leucine. Figure 5 The effect of treatment with acetyl-DL-leucine on lysosomal volume in fibroblasts from NPA, ML II, MPS IIIB, aspartylglucosaminuria, MLIIIA, and MPS VII patients is shown.

[0024] Figure 6A Survival curves representing mortality of wild-type and Sandhoff mice that were untreated or treated with acetyl-leucine are shown. Figures 6C-6H Bar crossing scores for Sandhoff model mice that were untreated and treated with acetyl-leucine are shown. Figure 7A Step cycle times for Sandhoff mice that were untreated and treated with acetyl-leucine, assessed at 12 weeks of age, are shown.

[0025] Figure 7B The effect of treatment with acetyl-DL-leucine on glycosphingolipid (GSL) levels in fibroblasts from GM2 gangliosidosis patients (Tay-Sachs, Sandhoff disease, and the AB variant of Tay-Sachs, respectively) is shown.

[0026] Figure 7C and 9B The effect of treatment with acetyl-DL-leucine on overall clinical severity score (CSS) and overall annual severity increment score (ASIS) for 10 NPC patients over time is shown.

[0027] ​Figures 8A-8C The effect of treatment with acetyl-DL-leucine on the CSS subscores over time is shown for each of the ten NPC patients. DETAILED DESCRIPTION

[0028] Acetyl-leucine in its racemic form (acetyl-DL-leucine) and its salts are effective in the treatment of vertigo of various origins, in particular Meniere's vertigo and vertigo of inflammatory (vestibular neuritis) or toxic origin. For example, acetyl-leucine is marketed in its racemic form by Pierre Fabre Medicament as an anti-vertigo drug under the trademark name Tanganil®. The clinical results reported by different authors with Tanganil® show an improvement of vertigo symptoms in more than 95% of cases, including the disappearance of vertigo attacks.

[0029] Acetyl-DL-leucine has been used in France since 1957 for the treatment of acute vertigo with a good safety profile, but its long-term safety in long-term use has not been determined. Its pharmacological and electrophysiological mode of action remains unclear, despite numerous hypotheses, including the stability of the membrane potential. (Vibert et al. (2001) Figure 9A ; 13(4): 735-48; Ferber-Viart et al. (2009) Figures 10A-10J ; 14(1): 17-25). FDG-µPET studies in a rat model of acute unilateral labyrinthectomy (Zwergal et al. (2016) Eur J Neurosci ; 221(1): 159-70) showed a significant effect of the L enantiomer, N-acetyl-L-leucine, on the vestibulo-cerebellar activation and the deactivation of the posterior lateral thalamus leading to a compensation of the posture (Gunther et al. (2015) Audiol Neurootol ; 10(3): e0120891). The symptomatic improvement of cerebellar ataxia using acetyl-DL-leucine was shown in a series of cases of cerebellar patients (Strupp et al. (2013) J Brain Struct Funct ; 260(10): 2556-61). Another series of cases did not find a benefit (Pelz et al. (2015) PLoS One ; 262(5): 1373-5). Quantitative gait analysis showed that acetyl-DL-leucine improved the temporal gait variability in patients with cerebellar ataxia (Schniepp et al. (2015) Neurol ; 3:8). In a one-month study involving 12 patients with Niemann-Pick disease type C (NPC), a symptomatic improvement of ataxia was shown (Bremova et al. (2015) J Neurol(2015) Neurology 85(16): 1368-75). Moreover, PET studies in ataxia patients given acetyl-DL-leucine showed increased metabolism in the midbrain and lower brainstem in responders (Becker-Bense et al. (2015) Cerebellum ) Neurology 85(16): 1368-75).

[0030] However, it was not known that acetyl-leucine treats LSD, which typically progresses over the course of years to decades. The present disclosure surprisingly demonstrates that acetyl-leucine, or a pharmaceutically acceptable salt thereof, can be used in methods of treating LSD in a subject in need thereof, e.g., by treating by delaying the onset of LSD or one or more symptoms of LSD that would otherwise be expected to occur according to typical disease progression, and / or by treating by delaying or reversing the progression (e.g., long-term) of LSD or one or more symptoms of LSD as compared to typical disease progression. These exemplary applications of the present disclosure, as well as other applications described herein, are completely unexpected because these benefits were not previously observed and could not be inferred, nor are they taught by the prior art. LSD is one of a heterogeneous group of genetic diseases that are typically characterized by the accumulation of undigested or partially digested macromolecules, leading to cellular dysfunction (e.g., increased lysosomal volume as compared to healthy subjects) and clinical abnormalities. As demonstrated by the examples, but without wishing to be bound by any particular theory, the present inventors have found, inter alia, that acetyl-leucine can improve cellular dysfunction (e.g., by reducing lysosomal volume to control values) and clinical abnormalities in subjects having LSD.

[0031] Accordingly, the present disclosure provides acetyl-leucine, or a pharmaceutically acceptable salt thereof, for use in a method of treating LSD or one or more symptoms of LSD in a subject in need thereof.

[0032] As used herein, "LSD" refers to any disorder involving dysfunction or disruption in the late endosomal / lysosomal system and accumulation of undigested or partially digested macromolecules. LSD can involve increased storage of lipids or non-lipid.

[0033] As used herein, "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, e.g., a domestic animal (e.g., a horse, cow, sheep, or pig), a pet (e.g., a cat, dog, rabbit, or guinea pig), a laboratory animal (e.g., a mouse or rat), or can be used in other veterinary applications. For example, the subject is a human.

[0034] As used herein, the singular forms "a," "an," and "the" include plural referents.

[0035] The terms“about” and“approximately” mean nearly the same as a reference number or value, including an acceptable degree of error for the amount measured in the given precision of the property or measurement. As used herein, the terms“about” and“approximately” are generally understood to encompass ±20% of the indicated amount, frequency, or value. Numerical values given herein are approximations unless otherwise stated, meaning that the term“about” or“approximately” can be inferred if not expressly stated.

[0036] The term“administering” as used herein means (1) providing, giving, dosing, and / or prescribing a composition according to the present disclosure by a health practitioner or an authorized agent or under the direction of the same, and (2) taking, consuming, or ingesting a composition according to the present invention by a patient or himself or herself.

[0037] All references to“acetyl- leucine” include pharmaceutically acceptable salts thereof, even if not expressly stated.

[0038] Acetyl-leucine can be in racemic form, meaning that the compound contains about equal amounts of enantiomers. Alternatively, it can be in enantiomeric excess of the L enantiomer or the D enantiomer. Acetyl-leucine can be in single enantiomeric form of the L enantiomer or the D enantiomer. In one embodiment, the single enantiomeric form is the L enantiomer. Racemic and enantiomeric forms can be included according to procedures known in the art.

[0039] References herein to“pharmaceutically acceptable salts” are any salt preparation suitable for pharmaceutical applications. 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-methylglucosamine, procaine, N-benzylphenethylamine, 1-p-chlorobenzyl-2-pyrrolidine-l'-ylmethylbenzimidazole, diethylamine, and other alkylamines, piperazine, tris(hydroxymethyl)aminomethane, and the like; alkali metal salts such as lithium, potassium, sodium, and the like; alkaline earth metal salts such as barium, calcium, magnesium, and the like; transition metal salts such as zinc, aluminum, and the like; other metal salts such as sodium hydrogen phosphate, disodium phosphate, and the like; inorganic acids such as hydrochloride, sulfate, and the like; organic acid salts such as acetate, lactate, malate, tartrate, citrate, ascorbate, succinate, butyrate, valerate, and fumarate, and the like.

[0040] Acetyl-leucine or a pharmaceutically acceptable salt thereof can be formulated and administered to a subject according to teachings known in the art. For example, acetyl-leucine, or a pharmaceutically acceptable salt thereof, can be formulated into a pharmaceutical composition. The pharmaceutical composition can include acetyl-leucine, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. References to a pharmaceutical composition can include the active agent alone or in the form of a pharmaceutical composition.

[0041] The pharmaceutical composition can take any of a wide variety of different forms, depending, inter alia, on the manner in which it is to be used. Thus, for example, it can be in the form of a powder, tablet, capsule, liquid, ointment, cream, gel, hydrogel, aerosol, spray, micellar solution, transdermal patch, liposome suspension or any other suitable form which can be administered to a human or animal in need of treatment.

[0042] Reference herein to a "pharmaceutically acceptable carrier" is any known compound or combination of known compounds which are known to those skilled in the art to be useful in formulating pharmaceutical compositions. It will be appreciated that the carrier of a pharmaceutical composition should be a substance that is tolerable by the subject to which it is to be administered.

[0043] In one embodiment, the pharmaceutically acceptable carrier can be a solid, and the composition can be in the form of a powder or tablet. The solid pharmaceutically acceptable carrier can include, but is not limited to, one or more substances which can also be a flavouring agent, a buffering agent, a lubricating agent, a stabilizing agent, a solubilizing agent, a suspending agent, a wetting agent, an emulsifying agent, a dye, a filler, a glidant, a compression aid, an inert binder, a sweetening agent, a preservative, a dye, a coating or a tablet disintegrant. The carrier can also be an encapsulating material. In a powder, the carrier can be a finely divided solid which is pre-mixed with the finely divided active agent according to the present application. In a tablet, the active agent can be mixed with a carrier having the necessary compression properties in the proper proportion and compressed into the desired shape and size. For example, the powder and tablet can 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, polyvinylpyrrolidine, low-melting wax and ion-exchange resins. In another embodiment, the pharmaceutically acceptable carrier can be a gel, and the composition can be in the form of a cream or the like.

[0044] The carrier can include, but is not limited to, one or more excipients or diluents. Examples of such excipients are gelatin, acacia, lactose, microcrystalline cellulose, starch, sodium starch glycolate, dicalcium phosphate, magnesium stearate, talc and colloidal silicon dioxide carbon and the like.

[0045] In another embodiment, the pharmaceutically acceptable carrier can be a liquid. In one embodiment, the pharmaceutical composition is in the form of a solution. Liquid carriers are used in preparing solutions, suspensions, emulsions, syrups, elixirs, and pressurized compositions. Acetyl-leucine can be dissolved or suspended in a pharmaceutically acceptable liquid carrier such as water, an organic solvent, a mixture of both or an oil or fat which is pharmaceutically acceptable. The liquid carrier can contain other suitable pharmaceutical additives such as solubilizers, emulsifiers, buffers, preservatives, sweeteners, flavoring agents, suspending agents, thickening agents, colorants, viscosity regulators, stabilizers, or osmo-regulators. Suitable examples of liquid carriers for oral and parenteral administration include water, partially

[0046] Liquid pharmaceutical compositions which are sterile solutions or suspensions can be utilized by, for example, intramuscular, intrathecal, epidural, intraperitoneal, intravenous, and especially subcutaneous injection. The active agent can be prepared as a sterile solid composition which can be dissolved or suspended at the time of administration using sterile water, saline, or other appropriate sterile injectable medium.

[0047] Compositions can be orally administered in the form of sterile solutions or suspensions containing other solutes or suspending agents, for example, enough saline or glucose to make the solution isotonic, bile salts, acacia, gelatin, sorbitan monoleate, polysorbate 80 (oleate esters of sorbitan with anhydrides of ethylene oxide) and the like. Compositions can also be administered in the form of liquid or solid compositions. 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 suitable for parenteral administration include sterile solutions, emulsions, and suspensions.

[0048] Acetyl-leucine and compositions comprising the same can alternatively be administered by inhalation (e.g., intranasally). Compositions can also be formulated for topical use. For example, a cream or ointment can be applied to the skin.

[0049] Acetyl-leucine can be incorporated into slow- or delayed-release devices. Such devices can for example be inserted on or under the skin, and the drug can be released over weeks or even months. Such devices can be advantageous when long-term treatment with acetyl-leucine according to the present disclosure is required, typically requiring frequent administration (e.g. at least daily administration).

[0050] In one embodiment, the pharmaceutical composition is in the form of a tablet. In a tablet, the active agent can be mixed with a carrier, e.g. a pharmaceutically acceptable carrier, having the necessary compression properties, in the proper proportion and compressed into a desired shape and size. A tablet can contain up to 99% by weight of the active agent.

[0051] For example, acetyl-leucine or a pharmaceutically acceptable salt thereof can be provided in a solid dosage form suitable for oral administration, in particular in the form of a tablet.

[0052] The pharmaceutical composition of a solid oral dosage form, e.g. a tablet, can be prepared by any method known in the pharmaceutical art. The pharmaceutical composition is typically prepared by mixing acetyl-leucine or a pharmaceutically acceptable salt thereof with conventional pharmaceutically acceptable carriers.

[0053] Tablets can be formulated as known in the art. For example Tanganil® ® Comprising wheat starch, pregelatinized corn (maize) starch, calcium carbonate and magnesium stearate as excipients. The same or similar excipients can for example be used with the present disclosure.

[0054] The composition of a 700 mg Tananil® tablet per tablet 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. The same tablet can for example be used with the present disclosure.

[0055] The present disclosure describes acetyl-leucine, including compositions and methods thereof, for treating LSD or one or more symptoms of LSD in a subject in need thereof. A subject in need can have a genetic, biochemical, or other similar identifiable marker of LSD. For example, a marker of LSD can be a cellular marker. A subject in need can be diagnosed with LSD. For example, a subject can have been diagnosed with LSD according to a genetic, biochemical, or other similar identifiable marker. A subject in need can be suspected of having or at risk of having LSD. For example, a subject can have a genetic predisposition for LSD (e.g., the subject can have one or more family members with LSD). A subject in need can be symptomatic (i.e., have one or more symptoms associated with LSD). A subject in need can be asymptomatic. It should be understood that the terms “symptomatic” and “asymptomatic” are used in reference to symptoms of LSD. A subject having a genetic, biochemical, or other similar identifiable marker of LSD, e.g., a subject diagnosed with LSD based on a genetic, biochemical, or other similar identifiable marker but without further symptoms of the disorder, is included within the scope of “asymptomatic” for the purposes of the present disclosure.

[0056] As used herein, “treating LSD or one or more symptoms of LSD” and the like refers to delaying the onset of LSD or one or more symptoms of LSD that would otherwise be expected to occur in accordance with typical disease progression, reducing the severity of LSD, or reducing the severity of or eliminating one or more existing symptoms associated with LSD, delaying the progression of LSD or one or more symptoms of LSD over time as compared to typical disease progression, and / or reversing the progression of LSD or one or more symptoms of LSD over time. “Treating LSD or one or more symptoms of LSD” can also be referred to as improving a biochemical marker of LSD.

[0057] As used herein, "typical disease progression," "generally expected disease progression," and the like refer to the typical or expected progression of an LSD, one or more symptoms associated with an LSD, or a biochemical marker of an LSD if the subject were not treated. Typical or expected disease progression can be based, for example, on a known scale, index, rating, or score, or other suitable test, for assessing the progression of an LSD, one or more symptoms associated with an LSD, or a biochemical marker of an LSD, such as those described by way of example herein. The scale, index, rating, or score, or other suitable test, can correspond to the overall progression of the disease or the progression of one or more symptoms associated with the disease. For example, typical or expected disease progression can be based on the typical or expected onset or severity of an LSD or a symptom or set of symptoms associated with an LSD. Typical or expected disease progression can be determined on a subject-by-subject basis, or can be based on what is generally observed or experienced by a collection of subjects affected by an LSD, such as a population or subpopulation of subjects. A subpopulation can include, for example, a subpopulation of the same gender, a subpopulation of the same or similar age, a subpopulation of the same or similar onset timing of one or more symptoms, and the like.

[0058] In one embodiment, "treating an LSD or one or more symptoms of an LSD" refers to delaying the onset of an LSD or one or more symptoms of an LSD that would otherwise be expected to occur according to typical disease progression. As used herein, "delaying the onset of an LSD or one or more symptoms of an LSD," and the like, refers to increasing the time until the onset of an LSD or one or more symptoms of an LSD, or preventing the onset thereof. For example, the onset can be said to be delayed when the time until the manifestation of an LSD or one or more symptoms of an LSD is at least 5% longer than would be observed according to typical disease progression. Further, for example, the time is observed to be increased by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%. In one embodiment, the subject is asymptomatic. Administration of acetyl-leucine can be initiated at a time when the subject is asymptomatic in order to delay the onset of an LSD or one or more symptoms of an LSD that would otherwise be expected to occur according to typical disease progression. In another embodiment, the subject is symptomatic. Administration of acetyl-leucine can be initiated at a time when the subject is symptomatic in order to delay the onset of one or more additional symptoms of an LSD that would otherwise be expected to occur according to typical disease progression. The subject in need can continue to receive acetyl-leucine treatment for the duration described herein. In one embodiment, the treatment prevents the onset of one or more symptoms of an LSD that would otherwise be expected to occur according to typical disease progression.

[0059] In one embodiment, "treating LSD or one or more symptoms of LSD" means reducing the severity of LSD, or reducing the severity of or eliminating one or more existing symptoms associated with LSD. Severity of LSD or existing symptoms can be assessed using a known scale, index, rating, or score, such as those described by way of example herein, or using another suitable test for assessing severity. For example, the scale, index, rating, or score, or other suitable test can correspond to overall severity of the disease or severity of one or more symptoms associated with the disease. In one embodiment, the treatment improves such assessment from a symptomatic patient's value or degree characteristic to an asymptomatic patient's value or degree characteristic.

[0060] In one embodiment, "treating LSD or one or more symptoms of LSD" means delaying progression of LSD or one or more symptoms associated with LSD, or reversing progression of LSD or one or more symptoms associated with LSD over time, as compared to typical disease progression. The time over which the treatment delays or reverses progression can be consistent with the treatment duration described herein. The treatment can delay or reverse progression over a duration of, for example, about 7 days or more, about 2 weeks or more, about 3 weeks or more, about 1 month or more, about 6 weeks or more, about 7 weeks or more, or about 2 months or more. For example, the treatment delays or reverses progression over a duration of about 3 months or more, about 4 months or more, about 5 months or more, or about 6 months or more. It can delay or reverse progression over a duration of, for example, about 1 year or more, about 2 years or more, about 3 years or more, about 4 years or more, about 5 years or more, or about 10 years or more. The treatment can delay or reverse progression of LSD or one or more symptoms associated with LSD over the lifetime of the patient.

[0061] In one embodiment, "treating LSD or one or more symptoms of LSD" refers to delaying the progression of LSD or one or more symptoms of LSD over time as compared to typical disease progression. As used herein, "delaying the progression of LSD or one or more symptoms of LSD over time," and the like, refers to slowing and / or stopping the progression of the disease or one or more symptoms of the disease over time (e.g., slowing and / or stopping the worsening or increasing severity of the disease or one or more symptoms of the disease). Disease progression can be determined, for example, using a known scale, index, rating, or score, such as those described by way of example herein, or using another suitable test for assessing progression. For example, the scale, index, rating, or score, or other suitable test, can correspond to the overall progression of the disease, or to the progression of one or more symptoms associated with the disease. In one embodiment, "delaying the progression of LSD or one or more symptoms of LSD" means that the severity value (e.g., overall severity or severity of one or more symptoms) of the disease in the subject, as determined by a scale, index, rating, or score, or other suitable test for assessing severity, does not increase meaningfully (e.g., at least remains substantially the same). In one embodiment, "delaying the progression of LSD or one or more symptoms of LSD" means preventing the subject from reaching a severity value, or increasing the time for the subject to reach a severity value (e.g., decreasing the rate of increase in severity), as compared to a value corresponding to typical disease progression, as determined by a known scale, index, rating, or score, or other suitable test for assessing progression. For example, progression can be said to be delayed when the time to reach a severity value is at least 5% longer than would be observed according to typical disease progression. Further, for example, the time is observed to increase by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100%. The time over which the treatment delays the progression of LSD or one or more symptoms of LSD can be consistent with the treatment duration described herein. In one embodiment, the treatment delays progression for at least about 3 months, at least about 4 months, at least about 5 months, or at least about 6 months. In another embodiment, the treatment delays progression for at least about 1 year, at least about 2 years, at least about 3 years, at least about 4 years, at least about 5 years, or at least about 10 years. The treatment can delay progression throughout the patient's life.

[0062] In one embodiment, "treating LSD or one or more symptoms of LSD" refers to reversing the progression of LSD or one or more symptoms of LSD over time. As used herein, "reversing the progression of LSD or one or more symptoms of LSD over time" and the like refer to halting and reducing the severity of the disease or one or more symptoms of the disease over time. Disease progression and severity are determined, for example, using a known scale, index, rating, or score, such as those described by way of example herein, or using another suitable test for assessing progression and severity. For example, the scale, index, rating, score, or other suitable test can correspond to overall progression and severity of the disease, or to progression and severity of one or more symptoms associated with the disease. In one embodiment, "reversing the progression of LSD or one or more symptoms of LSD over time" indicates that the subject's disease severity value (e.g., overall severity or severity of one or more symptoms) as determined by a known scale, index, rating, or score, and the like, or other suitable test for assessing severity, improves over time (i.e., shows a decrease in severity over time). The time over which treatment reverses the progression of LSD or one or more symptoms of LSD can be consistent with the treatment duration described herein. In one embodiment, treatment reverses the progression for at least about 3 months, at least about 4 months, at least about 5 months, or at least about 6 months. In another embodiment, treatment reverses the progression for at least about 1 year, at least about 2 years, at least about 3 years, at least about 4 years, at least about 5 years, or at least about 10 years. Treatment can reverse the progression throughout the patient's life.

[0063] In one embodiment, "treating LSD or one or more symptoms of LSD" refers to improving a biochemical marker of LSD in a subject (e.g., increasing the level of a storage metabolite or secondary biochemical changes resulting from the primary storage). Biochemical markers are signals of disease activity that can provide a continuous indication of disease severity and progression over time. In one embodiment, the biochemical marker is improved relative to a control value. In one embodiment, the biochemical marker is selected from the group consisting of increased lysosomal volume and increased levels of glycosphingolipids (GSLs). In one embodiment, the biochemical marker is increased lysosomal volume, and the treatment reduces lysosomal volume in the subject. In one embodiment, the biochemical marker is increased levels of glycosphingolipids (GSLs), and the treatment reduces GSL levels in the subject. In one embodiment, the treatment improves the biochemical marker over time. For example, in one embodiment, improving the biochemical marker over time means that the treatment improves the biochemical marker toward a control value over time, prevents progression of the biochemical marker over time, and / or delays progression of the biochemical marker over time as compared to typical disease progression. The time over which the treatment improves the biochemical marker can be consistent with the treatment duration described herein. In one embodiment, the treatment improves the biochemical marker for at least about 3 months, at least about 4 months, at least about 5 months, or at least about 6 months. In yet another embodiment, the treatment improves the biochemical marker for at least about 1 year, at least about 2 years, at least about 3 years, at least about 4 years, at least about 5 years, or at least about 10 years. The treatment can improve the biochemical marker throughout the patient's life.

[0064] A "symptom" of LSD includes any clinical or laboratory manifestation associated with LSD and is not limited to what a subject can feel or observe. Symptoms described herein include, but are not limited to, neurological symptoms and psychiatric symptoms. Examples of neurological symptoms include ataxia, other movement disorders such as hypokinesia, rigidity, tremor, or dystonia, central ocular motor disorders such as vertical and horizontal supranuclear gaze palsies, and neuro-psychological deficits such as dementia. Examples of psychiatric symptoms include depression, behavioral disturbances, or psychosis. The onset of symptoms can be from birth to adulthood.

[0065] The progression of LSD or one or more symptoms of LSD over time or through treatment can be monitored, for example, by monitoring at two or more time points using one or more known tests and comparing the results. Disease progression and / or severity can be assessed, for example, using the Ataxia Assessment Rating Scale (SARA), Spinocerebellar Ataxia Functional Index (SCAFI), International Collaborative Common Ataxia Rating Scale (ICARS), Brief Ataxia Rating Scale (BARS), Modified Disability Rating Scale (mDRS), EuroQol 5Q-5D-5L (EQ-5D-5L), Visual Analogue Scale (VAS), Wechsler Adult Intelligence Scale Revised Version (WAIS-R), Wechsler Intelligence Scale for Children-IV (WISC-IV), Montreal Cognitive Assessment (MoCA), or other appropriate tests. For certain LSDs, such as NPCs, specific scores have been developed and validated over the past few decades, such as the Clinical Severity Score (CSS) and the Annual Severity Increment Score (ASIS) (see Yanjanin et al., “Linear Clinical Progression, Independent of Age of Onset, in Niemann–Pick disease, Type C,” Neurology 153B:132–140), and the modified 6-Domain NP-C Disability Scale (mDRS score). For example, the severity of NPC patients can be quantified by assigning a CSS, which assesses various parameters of the disease (walking, seizures, eye movements, etc.) and gives a score of 5 for each parameter. A higher score equals a more severe degree. The ASIS quantifies the annual rate of change of CSS by dividing the CSS by the patient's age. In this respect, certain scores in these tests are characteristic of symptomatic LSD patients and those with clear disease progression and / or severity.

[0066] Thus, "treating LSD or one or more symptoms of LSD" can be equivalent, for example, to achieving an improvement in an assessment, such as SARA, SCAF1, ICARS, BARS, mDRS, EQ-5D-5L, VAS, WAIS-R, WISC-IV, CSS, and / or MoCA score, or another test result suitable for characterizing an LSD subject, as described herein. For example, in one embodiment, "reducing the severity of LSD, or reducing the severity of or eliminating one or more existing symptoms of LSD" means that the SARA, SCAF1, ICARS, BARS, mDRS, EQ-5D-5L, VAS, WAIS-R, WISC-IV, CSS, and / or MoCA score, or another test result suitable for assessing severity, such as a score or result, will change from a value characteristic of a symptomatic subject to a value characteristic of an asymptomatic subject. In another embodiment, "delaying progression of LSD or one or more symptoms of LSD" means that the SARA, SCAF1, ICARS, BARS, mDRS, EQ-5D-5L, VAS, WAIS-R, WISC-IV, CSS, and / or MoCA score, or another suitable test result for assessing progression, of a subject does not increase significantly (e.g., at least remains essentially constant). In yet another embodiment, "delaying progression of LSD or one or more symptoms of LSD" means preventing a subject's SARA, SCAF1, ICARS, BARS, mDRS, EQ-5D-5L, VAS, WAIS-R, WISC-IV, CSS, and / or MoCA score, or another suitable test result for assessing progression, from reaching a value comparable to that typically seen in disease progression, or increasing the time it takes to reach that value. In another embodiment, "reversing progression of LSD or one or more symptoms of LSD over time" means that a subject's SARA, SCAF1, ICARS, BARS, mDRS, EQ-5D-5L, VAS, WAIS-R, WISC-IV, CSS, and / or MoCA score, or another suitable test result for assessing progression, improves over time (i.e., shows a decrease in severity over time).

[0067] For example, for assessing the overall nervous system status, the mDRS, a four-domain scale (ambulation, manipulation, language, and swallowing) can be applied. Cerebellar function can be assessed using the SARA, a eight-item clinical rating scale (gait, stance, sitting posture, speech, fine motor function, and praxis; range 0-40, where 0 is the best neurological state and 40 is the worst), SCAFI, 9-hole peg test (9HPT), and the number of “PATA” repetitions in 10 seconds. Subjective impairment and quality of life can be assessed using the EQ-5D-5L assessment questionnaire and VAS. For assessing eye movement function, the three-dimensional video-oculography (EyeSeeCam) can be used to measure the peak velocity of saccades, gain of smooth pursuit, peak slow-phase velocity of gaze-evoked nystagmus (gaze holding function), peak slow-phase velocity of optokinetic nystagmus, and gain of the vestibulo-ocular reflex. For assessing cognitive status, the WAIS-R or WISC-IV and MoCA can be used to assess different cognitive domains, including attention and concentration, executive function, memory, language, visual-spatial skills, conceptual thinking, calculation, and orientation, with a maximum of 30 points and a cutoff score of 26. The skilled person will know how to perform these tests and other such tests.

[0068] Acetyl-leucine or a pharmaceutically acceptable salt thereof can be administered, for example, in a dose of about 500 mg to about 15 g per day or about 500 mg to about 10 g per day, for example about 1.5 g to about 10 g per day, optionally by the solid oral or liquid oral route. Acetyl-leucine or a pharmaceutically acceptable salt thereof can be administered, for example, in a dose according to Tanganil® ® , for which the prescription for adults is a dose of 1.5 g to 2 g per day, 3-4 tablets, divided into two doses.

[0069] If one enantiomer is administered, the dose can be reduced accordingly. For example, if only acetyl-L-leucine or only acetyl-D-leucine is administered, the dose can be about 250 mg to about 15 g per day, about 250 mg to about 10 g per day, or about 250 mg to about 5 g per day, for example about 0.75 g to about 5 g per day.

[0070] In one embodiment, the dose administered is 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. 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 more than about 1.5 g per day, but less than about 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, or 5 g per day. In one embodiment, the dose is about 4 g to about 6 g per day. In one embodiment, the dose is about 4 g to about 5 g per day. In one embodiment, the dose is about 4.5 g per day. In one embodiment, the dose is about 5 g per day. In one embodiment, these doses are administered in solid oral dosage forms, in particular tablets. In another embodiment, these doses are for acetyl- leucine when in racemic form. When there is an enantiomeric excess, the dose of acetyl-leucine can be lower than those described herein, e.g., about 50% lower. Thus, the present disclosure also expressly encompasses the above dose ranges when halved.

[0071] The total daily dose can be distributed over multiple administrations, i.e. the administration can be done in two or more portions each day to achieve the total daily dosage. For example, the number of tablets needed to provide the total daily dose of acetyl-leucine can be divided into two administrations (e.g. in the morning and in the evening) or three administrations (e.g. in the morning, at noon and in the evening). Each administration can be suitably administered with or without food. For example, acetyl-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 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, a total daily dose of 4.5 grams of acetyl-DL-leucine can be administered as 3 tablets of Tanganil (or equivalent) before, during or after breakfast, another 3 tablets before, during or after lunch, and a further 3 tablets before, during or after dinner. ® (Or equivalent), another 3 tablets before, during or after lunch, and a further 3 tablets before, during or after dinner.

[0072] Acetyl- leucine administration according to the present disclosure can be initiated prior to or after the subject is found to have a genetic, biochemical, or other similarly identifiable marker of LSD, e.g., in the former case, when the subject is suspected of having or at risk of having LSD. Administration can be initiated at or around the time the subject is found to have a genetic, biochemical, or other similarly identifiable marker of LSD. Likewise, administration can be initiated prior to, at or around the time of, or after the subject is diagnosed with LSD, or prior to, at or around the time of, or after the subject is found to have a genetic, biochemical, or other similarly identifiable marker of LSD. Acetyl-leucine administration can be initiated while the subject is symptomatic or asymptomatic. In particular, one of the advantages of acetyl-leucine treatment according to the present disclosure is that acetyl-leucine administration can be initiated as early as the subject is found to have a genetic and / or biochemical marker of LSD but prior to the subject showing symptoms of LSD (symptoms other than the genetic and / or biochemical markers, i.e., the subject is asymptomatic) or prior to the subject showing signs considered markers of the disease. Treatment can delay the onset of LSD or one or more symptoms associated with LSD, as described herein. Treatment can also last for a duration as described herein.

[0073] As discussed herein, an advantage of acetyl-leucine treatment according to the present disclosure is that acetyl-leucine can be administered over a long period of time to, e.g., delay or even reverse the progression of LSD or one or more symptoms of LSD in the subject as compared to typical disease progression. The duration of treatment can be, e.g., more than about 7 days, more than about 2 weeks, more than about 3 weeks, more than about 1 month, more than about 6 weeks, more than about 7 weeks, or more than about 2 months. In one embodiment, it is more than about 3 months, more than about 4 months, more than about 5 months, or more than about 6 months. The duration of treatment can be more than about 1 year, more than about 2 years, more than about 3 years, more than about 4 years, more than about 5 years, or more than about 10 years. The duration of treatment can be for the life of the patient.

[0074] Any and all combinations of dosage form, dosage, dosing regimen, and duration of treatment are contemplated and encompassed by the present invention. In one embodiment, the dosage is from about 4 g to about 10 g per day, administered in one, two, or three administrations per day, for a duration of treatment of about 2 months or more. In another embodiment, the dosage is more than 4 grams but not more than 5 grams per day, administered in one, two, or three administrations per day, for a duration of treatment of about 6 months or more. The dosage form can be a solid oral dosage form, in particular a tablet.

[0075] The pharmaceutical compositions can be used as monotherapy (e.g., using the active agent alone) to treat LSD in a subject. Alternatively, the pharmaceutical compositions can be used as an adjunct or in combination with other known therapies, e.g., for treating LSD in a subject.

[0076] All LSDs, which can be classified in various ways, are within the scope of the present disclosure. In one embodiment, the LSD is selected from any one of a glycogen storage disease, a mucopolysaccharide storage disease, a mucolipid storage disease, an oligosaccharide storage disease, a lipid storage disease, a sphingolipid storage disease, and a lysosomal transport disease.

[0077] The sphingolipid storage disease can be selected from any one of Niemann Pick disease type A / B, Gaucher disease types I, II, and III, Krabbe disease, Fabry disease, Schindler disease, GM1 gangliosidosis, Morquio B disease, GM2 gangliosidosis, metachromatic leukodystrophy, Farber disease, multiple sulfatase deficiency, lysosomal acid lipase deficiency, and galactosidosis. In one embodiment, the sphingolipid storage disease is selected from Niemann Pick disease type A, GM1 gangliosidosis, Tay Sachs disease, AB variant of Tay Sachs disease, and Sandhoff disease.

[0078] The mucolipid storage disease can be selected from any one of Mucolipidosis type I, Mucolipidosis type II, Mucolipidosis type III, and Mucolipidosis type IV. In one embodiment, the mucolipid storage disease is Mucolipidosis type II or Mucolipidosis type III.

[0079] The mucopolysaccharide storage disease can be selected from any one of MPS IH, MPS IH-S, MPS IS, MPS IIA, MPS IIB, MPS IIIA-D, MPS IVA, MPS VI, MPS VII, and MPS IX. In one embodiment, the mucopolysaccharide storage disease is MPS III or MPS VII. In one embodiment, the mucopolysaccharide storage disease is MPS IIIB.

[0080] The oligosaccharide storage disease can be selected from any one of β-mannosidosis, a-fucosidosis, and aspartylglucosaminuria. In one embodiment, the oligosaccharide storage disease is aspartylglucosaminuria.

[0081] The lipid storage disease can be selected from any one of Niemann Pick disease type C, Niemann Pick disease type D, neuronal ceroid lipofuscinosis (including types I-X), and Wolman disease. In one embodiment, the lipid storage disease is Niemann Pick disease type C.

[0082] The glycogen storage disorder can be selected from Pompe disease, late-onset Pompe disease and Danon disease.

[0083] The lysosomal transport disease can be selected from cystinosis, pycnodysostosis, sialic acid storage disease and infantile free sialic acid storage disease.

[0084] The LSD can be a primary lysosomal hydrolase deficiency, a post-translational processing deficiency of a lysosomal enzyme, a transport deficiency of a lysosomal enzyme, a protection deficiency of a lysosomal enzyme, a soluble non-enzyme lysosomal protein deficiency, a transmembrane (non-enzyme) protein deficiency or an unclassified deficiency.

[0085] In one embodiment, the LSD is selected from a primary lysosomal hydrolase deficiency. Primary lysosomal hydrolase deficiencies include, but are not limited to, Tay-Sachs disease (beta-hexosaminidase A deficiency), Sandhoff disease (beta-hexosaminidase A+B deficiency), Fabry disease (alpha-galactosidase A deficiency), Krabbe disease (beta-galactosylceramidase deficiency), Niemann Pick disease types A and B (sphingomyelinase deficiency), Metachromatic leukodystrophy (arylsulfatase A deficiency), MPS IH (Hurler syndrome; alpha-iduronidase deficiency), MPS IS (Scheie syndrome; alpha-iduronidase deficiency), MPS IH-S (Hurler-Scheie syndrome; alpha-iduronidase deficiency), MPS II (Hunter syndrome; iduronate sulfatase deficiency), MPS IIIA (Sanfilippo A syndrome; heparan acetyl esterase deficiency), MPS IIIB (Sanfilippo B syndrome; acetyl alpha-glucosaminidase deficiency), MPS IIIC (Sanfilippo C syndrome; acetyl CoA:alpha-glucosaminidase N-acetyltransferase deficiency), MPS IIID (Sanfilippo D syndrome; N-acetylglucosamine-6-sulfatase deficiency), MPS IV A (Morquio A disease; acetyl galactosamine-6-sulfatase deficiency), MPS IVB (Morquio B disease; beta-galactosidase deficiency), MPS V (renamed MPS IS), MPS VI (Maroteaux Lamy Syndrome; acetyl galactosamine-4-sulfatase (arylsulfatase B) deficiency), MPS VII (Sly syndrome; beta-glucuronidase deficiency), MPS IX (hyaluronidase deficiency), Wolman / Cholesterol ester storage disease (WD; acid lipase deficiency), Pompe disease (type II; alpha 1,4-glucosidase deficiency), aspartylglycosaminuria (glycosylasparaginase deficiency), fucosidosis (alpha-fucosidase deficiency), alpha-mannosidosis (alpha-mannosidase deficiency), beta-mannosidosis (beta-mannosidase deficiency), Schindler disease (N-acetylgalactosaminidase deficiency), sialidosis / ML I (alpha-neuraminidase deficiency), infantile neuronal ceroid lipofuscinosis (CLN1; palmitoyl protein thioesterase deficiency), late infantile neuronal ceroid lipofuscinosis (CLN2; carboxypeptidase deficiency), early infantile GM1 gangliosidosis, late infantile GM1 gangliosidosis, adult infantile GM1 gangliosidosis, Gaucher disease type 1 (non-neuronopathic), Gaucher disease types 2 / 3 (neuronopathic), neuronal ceroid lipofuscinosis type 4 (CLN4; Kufs disease; adult NCL; palmitoyl protein thioesterase-1 deficiency (type A);Cathepsin F deficiency (B-type), Neuronal ceroid lipofuscinosis type 10 (CLN10; congenital cathepsin D deficiency), Dense Osteogenesis Imperfecta (cathepsin K deficiency), Infantile Pompe disease, Late-onset Pompe disease, Fabry disease (Fabry's lipogranulomatosis; ceramidase deficiency; fibrochondrogenesis; lipogranulomatosis), and Galactosialidosis (protective protein cathepsin A deficiency, PPCA deficiency). In one embodiment, the primary lysosomal hydrolase deficiency is selected from the group consisting of Tay-Sachs disease, Sandhoff disease, Niemann-Pick disease type A, Niemann-Pick disease type B, neuronal ceroid lipofuscinosis, Gaucher disease, Fabry disease, Krabbe disease, GM1 gangliosidosis, GM2 gangliosidosis, metachromatic leukodystrophy, and Fabry disease. In one embodiment, the primary lysosomal hydrolase deficiency is selected from the group consisting of Tay-Sachs disease, Sandhoff disease, Niemann-Pick disease type A, Niemann-Pick disease type B, and GM1 gangliosidosis.

[0086] In one embodiment, the LSD is selected from the group consisting of post-translational processing deficiencies of lysosomal enzymes. Post-translational processing deficiencies of lysosomal enzymes include, but are not limited to, multiple sulfatase deficiency (MSD; deficiency of multiple sulfatases), MLII (I-cell disease; N-acetylglucosamine phosphotransferase deficiency), and MLIII (pseudo-Hurler polydystrophy; N-acetylglucosamine phosphotransferase deficiency).

[0087] In one embodiment, the LSD is selected from the group consisting of lysosomal enzyme transport deficiencies. Lysosomal enzyme transport deficiencies include, but are not limited to, mucolipidosis type II (I-cell disease; N-acetylglucosamine phosphotransferase deficiency), mucolipidosis type ID (pseudo-Hurler polydystrophy; N-acetylglucosamine phosphotransferase deficiency), and mucolipidosis type III C.

[0088] In one embodiment, the LSD is a lysosomal enzyme protection deficiency. Lysosomal enzyme protection deficiencies include, but are not limited to, galactosialidosis (protective protein cathepsin A (PPCA) deficiency).

[0089] In one embodiment, the LSD is a soluble non-enzyme lysosomal protein deficiency. Soluble non-enzyme lysosomal protein deficiencies include, but are not limited to, GM2 activator protein deficiency (AB variant), Niemann-Pick disease type C2 (NPC2), sphingolipid activator protein (SAP) deficiency.

[0090] In one embodiment, the LSD is a transmembrane (non-enzyme) protein deficiency. Transmembrane (non-enzyme) protein deficiencies include, but are not limited to, Danon disease (lysosome-associated membrane protein 2 (LAMP2) deficiency), NPC (NPC1 deficiency), cystinosis (cystinosin deficiency), infantile free sialic acid storage disease (ISSD; sialic acid transporter deficiency), Salla disease (free sialic acid storage disease; sialic acid transporter deficiency), juvenile neuronal ceroid lipofuscinosis (CLN3, Batten disease), adult neuronal ceroid lipofuscinosis (Kufs disease; adult NCL; palmitoyl protein thioesterase-1 deficiency (type A); cathepsin F deficiency (type B)), neuronal ceroid lipofuscinosis (NCL) (CLN6, CLN7, and LN8), and mucolipidosis type IV (mucolipidin deficiency). In one embodiment, the LSD is Niemann Pick disease type Cl or Niemann Pick disease type C2.

[0091] In one embodiment, the LSD is an unclassified deficiency. Unclassified deficiencies include, but are not limited to, neuronal ceroid lipofuscinosis (NCL) (CLN5 and CLN9).

[0092] The LSD to be treated by the compositions and methods of the present application can be any one of neuronal ceroid lipofuscinosis, primary sphingolipidoses (i.e., Gaucher disease, Fabry disease, GM1, GM2 gangliosidoses, Krabbe disease, and metachromatic leukodystrophy (MLD)), Fabry disease, and multiple sulfatase deficiencies. In one embodiment, the LSD has significant central nervous system (CNS) involvement. For example, the LSD can be selected from the group consisting of NPC, Tay Sachs disease, Sandhoff disease, GM1 gangliosidosis, or Fabry disease.

[0093] In one embodiment, the LSD is Niemann Pick disease type A. In another embodiment, the LSD is Niemann Pick disease type B. In another embodiment, the LSD is Niemann Pick disease type C (Cl or C2). Niemann Pick disease is a heterogeneous group of autosomal recessive LSDs. Common cellular features include abnormal sphingomyelin (SM) storage in mononuclear phagocytes and parenchymal tissues, and hepatosplenomegaly. In three major subgroups (A-C), NPC (previously classified as NPC and NPD, now considered a single disease) is classified as a fatal neuroendocrine LSD caused by an accumulation of unesterified cholesterol induced by abnormal intracellular cholesterol trafficking in the late endosomal / lysosomal compartment. Outside the CNS, cellular features of NPC include abnormal accumulation of unesterified cholesterol and other lipids (such as GSLs) within the late endosomal / lysosomal compartment. In contrast, there is no net elevation of cholesterol in the CNS (although it does have an altered distribution) but there are highly elevated levels of GSLs. Progressive neurodegeneration is particularly characterized by the continuous degeneration of GABAergic Purkinje neurons of the cerebellum, which parallels the onset and progression of cerebellar ataxia and other aspects of neurological dysfunction that occur during NPC. Genetic studies have shown that NPC disease is caused by mutations in either the Npc1 or Npc2 genes. The precise mechanistic link between these two genes remains unknown, and the functional roles of these proteins remain enigmatic. NPC1 encodes a multi-membrane spanning protein of the limiting membrane of the late endosome / lysosome, while NPC2 is a soluble cholesterol-binding protein of the lysosome. When NPC1 is inactivated, sphingosine is the first lipid to be stored, suggesting that NPC1 plays a role in the transport of sphingosine in the lysosome, which is normally produced as part of sphingolipid catabolism. Elevated sphingosine in turn leads to a defect in calcium entry into the acidic store, resulting in a greatly reduced release of calcium from this compartment. This in turn can prevent late endosome-lysosome fusion, which is a calcium-dependent process, and leads to a secondary accumulation of lipids (cholesterol, sphingomyelin, and glycosphingolipids) that are cargo transported through the late endocytic pathway. Other secondary consequences of inhibiting NPC1 function include defective endocytosis and failure to clear autophagic vacuoles. It has been shown that the NPC1 / NPC2 cellular pathway is targeted by pathogenic mycobacteria to facilitate their survival in the late endosome.

[0094] Tay Sachs disease is a fatal genetic disease of lipid metabolism characterized particularly in CNS tissues, which is caused by a deficiency of the A isozyme of β-hexosaminidase. Mutations in the HEXA gene, which encodes the alpha subunit of β-hexosaminidase, result in the A isozyme deficiency. Tay Sachs is the prototype of a group of disorders, GM2 gangliosidoses, characterized by a defect in the degradation of GM2 ganglioside. GM2 ganglioside (monosialo ganglioside 2) has begun to accumulate in neurons during fetal life.

[0095] Sandhoff disease is caused by a deficiency in the A and B (basic) isozymes of beta- hexosaminidase. Mutations in the HEXB gene, which encodes the beta subunit of beta- hexosaminidase, result in deficiency of the B isozyme.

[0096] GM1 gangliosidosis is caused by a deficiency in beta-galactosidase, which results in lysosomal accumulation of GM1 ganglioside (monosialylated ganglioside 1).

[0097] Fabry disease is caused by a deficiency in alpha-galactosidase, which results in lysosomal accumulation of ceramide trihexoside.

[0098] In one embodiment, the LSD is selected from the group consisting of Tay-Sachs disease, AB variant of Tay-Sachs disease, Sandhoff disease, Niemann Pick disease type A, Mucolipidosis type II, Mucolipidosis type III, MPS III, MPS VII, GM1 gangliosidosis, and aspartylglucosaminuria. In one embodiment, the LSD is Sandhoff disease. In one embodiment, the LSD is Tay-Sachs disease. In one embodiment, the LSD is the AB variant of Tay-Sachs disease. In one embodiment, the LSD is Mucolipidosis type II. In one embodiment, the LSD is Mucolipidosis type III. In one embodiment, the LSD is GM1 gangliosidosis. In one embodiment, the LSD is MPS III. In one embodiment, the LSD is MPS VII. In one embodiment, the LSD is Niemann Pick disease type A. In one embodiment, the LSD is aspartylglucosaminuria.

[0099] In one embodiment, the LSD is not Niemann Pick disease. In one embodiment, the LSD is not Niemann Pick disease type C.

[0100] In one embodiment, acetyl-leucine or a pharmaceutically acceptable salt thereof treats weight loss, gait deterioration, and / or deterioration of motor function associated with Niemann Pick disease (e.g., Niemann Pick disease type C or A) or Mucolipidosis type II. For example, acetyl-leucine or a pharmaceutically acceptable salt thereof can delay the onset of, reduce the severity of, or eliminate it, or delay or reverse its progression, of weight loss, gait deterioration, and / or deterioration of motor function associated with Niemann Pick disease (e.g., Niemann Pick disease type C or A) or Mucolipidosis type II. In one embodiment, the weight loss, gait deterioration, and / or deterioration of motor function is associated with Niemann Pick disease type A or Mucolipidosis type II.

[0101] In one embodiment, acetyl- leucine or a pharmaceutically acceptable salt thereof treats gait deterioration, motor function deterioration, and / or motor decline associated with Sandhoff disease. For example, acetyl-leucine or a pharmaceutically acceptable salt thereof can delay onset, reduce severity, or eliminate gait deterioration, motor function deterioration, and / or motor decline associated with Sandhoff disease, or delay or reverse progression thereof.

[0102] In one embodiment, acetyl- leucine or a pharmaceutically acceptable salt thereof treats coordination decline, tremor, motor decline, cognitive impairment, and / or gait deterioration associated with Tay Sachs disease. For example, acetyl-leucine or a pharmaceutically acceptable salt thereof can delay onset, reduce severity, or eliminate coordination decline, tremor, motor decline, cognitive impairment, and / or gait deterioration associated with Tay Sachs disease, or delay or reverse progression thereof.

[0103] Also provided is a method of treating LSD or one or more symptoms of LSD in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of acetyl-leucine or a pharmaceutically acceptable salt thereof.

[0104] A "therapeutically effective amount" of an agent is any amount which, when administered to a subject, is needed to produce a desired effect, which for the present disclosure can be therapeutic and / or prophylactic. Dosages can be determined in accordance with a variety of parameters, such as the particular form of acetyl-leucine used; the age, weight, and condition of the patient being treated; the type of disease; the route of administration; and the desired treatment regimen. A physician will be able to determine a desired route of administration and dosage for any particular patient. For example, a daily dose can 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.

[0105] Also disclosed is a kit for treating LSD in a subject in need thereof (e.g., a subject having, suspected of having, or at risk of having LSD), comprising a means for diagnosing or prognosing LSD, and acetyl-leucine or a pharmaceutically acceptable salt thereof.

[0106] The means for diagnosing or prognosing LSD can include a specific binding agent, a probe, a primer, a primer pair or primer combination, an enzyme, or an antibody, including antibody fragments, capable of detecting or aiding in the detection of LSD as defined herein. The kit can comprise LysoTracker® ® which is a fluorescent marker, and is commercially available from Invitrogen, also from Lonza. LysoTracker® can be blue, blue-white, yellow, green, or red.

[0107] The kit also comprises acetyl-leucine or a pharmaceutically acceptable salt thereof as defined herein. The kit can further comprise a buffer or aqueous solution. The kit can further comprise instructions for using acetyl-leucine or a pharmaceutically acceptable salt thereof in the methods of the application.

[0108] In yet another embodiment, acetyl-leucine or a pharmaceutically acceptable salt thereof is disclosed for use in a method of providing neuroprotection in a subject having, suspected of having, or at risk of having LSD.

[0109] As used herein, "neuroprotection" and synonyms thereof refer to the prevention of neurodegeneration, slowing and / or reversing the progression of neurodegeneration, including but not limited to progressive loss of neuronal structure, progressive loss of neuronal function, and / or progressive neuronal death. Providing neuroprotection can result in delaying the onset of LSD or one or more symptoms of LSD that would otherwise be expected to occur in accordance with typical disease progression, reducing the severity of LSD, or reducing the severity of or eliminating one or more preexisting symptoms associated with LSD, delaying the progression of LSD or one or more symptoms of LSD over time as compared to typical disease progression, and / or reversing the progression of LSD or one or more symptoms of LSD over time. The time at which neuroprotection is provided can coincide with the duration of treatment described herein. Further, for example, the treatment provides neuroprotection for a duration of, e.g., greater than about 7 days, greater than about 2 weeks, greater than about 3 weeks, greater than about 1 month, greater than about 6 weeks, greater than about 7 weeks, or greater than about 2 months. The treatment can provide neuroprotection for a duration of greater than about 3 months, greater than about 4 months, greater than about 5 months, or greater than about 6 months. In another embodiment, it provides neuroprotection for a duration of, e.g., greater than about 1 year, greater than about 2 years, greater than about 3 years, greater than about 4 years, greater than about 5 years, or greater than about 10 years. The treatment provides neuroprotection over the course of the patient's life.

[0110] As demonstrated by the examples, the inventors of the present application believe that acetyl-leucine acts as a neuroprotective agent and thus inhibits neurodegeneration that would otherwise be expected to occur.

[0111] In one embodiment, there is a method of providing neuroprotection in a subject having, suspected of having, or at risk of having LSD, the method comprising administering to the subject a therapeutically effective amount of acetyl-leucine or a pharmaceutically acceptable salt thereof.

[0112] Also disclosed is a kit for providing neuroprotection in a subject having, suspected of having, or at risk of having LSD, the kit comprising a means for diagnosing or predicting LSD, and acetyl-leucine or a pharmaceutically acceptable salt thereof.

[0113] The present disclosure also includes the use of acetyl-leucine, or a pharmaceutically acceptable salt thereof, as a neuroprotective agent in a subject having, suspected of having, or at risk of having LSD.

[0114] All features described herein (including any accompanying claims, abstract and drawings) and / or all of the steps of any method so disclosed can be combined in any combination, except combinations where at least some of such features and / or steps are mutually exclusive.

[0115] EMBODIMENTS

[0116] The application will now be further explained in the following examples, which demonstrate the effect of acetyl-leucine in treating LSD in a subject and providing neuroprotection in said subject.

[0117] EMBODIMENT 1

[0118] In vivo mouse studies - methods

[0119] Abstract EAN

[0120] This study takes advantage of a true mouse model of NPC, i.e. Am J Med Genet Part B - / - (BALB / cNctr- Mouse model m1N / J ) mice, which are null for the NPC1 protein and show all hallmarks of the clinical disease (Loftus, 1997).

[0121] This mutant line arises spontaneously and has a lifespan of 10-14 weeks, thus having a more acute course of the disease than the vast majority of patients. Mutant mice have been successfully developed, not only for determining the individual genesis of the disease and potential pathogenic mechanisms, but also for evaluating experimental therapies. Analyses using these mice have been performed at the whole-animal, cellular and molecular levels (Baudry, 2003; Smith, 2009; Cologna, 2014; Cologna, 2012). It is the most thoroughly studied animal model of NPC.

[0122] Before approximately 4-5 weeks of age, Npc1 - / - Mice do not have overt signs of disease behavior that distinguish them from wild-type littermates. The first signs of behavioral deficits, such as tremors and ataxic gait, appear at 5-6 weeks; by 7-8 weeks, the deficits in motor coordination become more pronounced, and by 9-10 weeks, ataxia progresses, accompanied by weight loss and exacerbation of poor coat condition due to difficulties in feeding and drinking water (human end points are applied) (Smith, 2009).

[0123] Wild type Npc1 - / - Littermate mice were used as controls.

[0124] Npc1

[0125] From weaning (3 weeks of age), Npc1 - / - Mice groups and Treatment procedure + / + Mice groups were treated with 0.1 g / kg acetyl-DL-leucine, which was mixed in the mice chow. Separate Npc1 - / - and Npc1 + / + Mice groups were treated for 9 weeks, as controls.

[0126] Npc1

[0127] By simple observation of 9-week-old mice, the coat condition of mice treated and not treated with acetyl-DL-leucine was compared. Npc1 - / - The coat condition of mice was evaluated by simple observation.

[0128] Coat condition

[0129] Animals were weighed twice a week. The body weight of all mice in each group was averaged (mean) and compared.

[0130] Npc1

[0131] Gait analysis was performed on 8-week-old mice using the CatWalk® system according to the manufacturer's instructions (Noldus, Nottingham, UK). Five runs were recorded for each animal. ® 15.0

[0132] The CatWalk® parameters measured were: 1. Stance mean: average duration (s) of the paw contact with the glass plate; 2. Step cycle: duration (s) between two consecutive contacts of the same paw; 3. Duty cycle: percentage of time the paw is in contact with the plate compared to the time to complete a step cycle; 4. Step sequence (AB): percentage of time spent walking in the LF-RH-RF-LH alternating pattern (LF: left front; RH: right rear; RF: right front; LH: left rear); 5. Step frequency: number of steps per second during the trial; 6. Diagonal support: percentage of time the diagonal paws are in contact with the glass plate (RF & LH or RH & LF).

[0133] Body weight data

[0134] Motor function analysis was performed on 8-week-old and 9-week-old mice using the OpenField Activity Monitor, following the manufacturer's instructions (Linton Instruments, Amlogger Software). Each mouse was placed in a plastic cage with bedding and analyzed for 5 minutes. Rears were manually calculated.

[0135] The measured motion function parameters are: 1. Central upright posture: Mice stand upright on their hind legs without support; 2. Upright posture: Mice stood upright on their hind legs, both with and without cage wall support; 3. Activity: Regular exercise for animals, including walking; 4. Front-to-back (FR) counting: the movement of the animal from the front to the back of the cage; 5. Activity Duration: The duration of the activity (s / min), regardless of movement; 6. Movement time: Duration of movement (s / min); 7. Standing time: The duration of any upright position.

[0136] result

[0137] Gait analysis

[0138] Motor function analysis Showing untreated Coat condition - / - Age-matched littermate mice. They were observed to have difficulty feeding and drinking water. Figure 1B - / - Mice had poor coat condition at 9 weeks of age (see [reference]). Npc1 ).

[0139] In stark contrast, Npc1 This shows the results of treatment with acetyl-DL-leucine from weaning onwards. Figure 1B - / - Mice. Treated with acetyl-DL-leucine Figure 1A - / - The mice have smooth, glossy fur, reminiscent of wild-type mice. Npc1 + / + )Livestock mice (see Npc1 ).

[0140] Npc1

[0141] from Figure 1A It can be seen from the wild type (Body weight data + / + ) Mice gradually gained weight during the study, i.e. from 3 to 10 weeks of age. In addition, Figure 2A The average weight of each group of mice at each time point is shown Npc1 - / - Untreated, n = 1 ; Figure 2A - / - Acetyl-DL-leucine 0.1 g / kg, n = 3; Npc1 + / + Untreated, n = 3; Npc1 + / + Acetyl-DL-leucine 0.1 g / kg, n = 2).

[0142] Treatment with acetyl-DL-leucine had no significant effect on this weight gain.

[0143] Npc1 - / - Mice initially gained weight, mainly with Npc1 + / + In the same way as the control group. However, then Npc1 - / - Mice started to lose weight from six weeks. At the end of the study (10 weeks of age), the mice had almost as little weight as at four weeks of age.

[0144] Treatment with acetyl-DL-leucine delayed these weight loss symptoms by two weeks compared to the untreated group.

[0145] Npc1 The weight of each group of mice with and without acetyl-DL-leucine treatment is shown Npc1 - / - Comparison of the weight change of the mice. In particular, Figure 2B Only for Npc1 - / - The weight change (%) of each group of mice at each time point is shown for the mice. From this figure, the beneficial effect of acetyl-DL-leucine treatment delaying weight loss is clearly visible.

[0146] Figure 2B

[0147] The results of the gait analysis are shown in Figure 3. The diagonal support, step frequency and step sequence data are shown in Npc1 respectively. Gait analysis and 3E The forepaw (FP) data Figures 3A-3C show the average stance and step cycle in Figure 3D show the duty cycle in Figure 3D and 3G The hindpaw (HP) data Figure 3Eaverage and stride cycle in the stance phase; Figure 3F Data are expressed as mean ± SEM. For untreated Figure 3F + / + , n = 3, for treated Figure 3G + / + , n = 2, for untreated Npc1 - / - , n = 1 (thus no statistical analysis was performed), for treated Npc1 - / - , n = 3.

[0148] The first bar in each graph shows the gait characteristics of wild-type ( Npc1 + / + ) mice.

[0149] The second bar in each graph shows the gait characteristics of wild-type ( Npc1 + / + ) mice treated with acetyl-DL-leucine. There was no significant difference in gait characteristics between these mice and untreated littermates.

[0150] The third bar in each graph shows the gait characteristics of Npc1 - / - mice. Overall, these mice exhibited a poor gait compared to Npc1 + / + mice. The mice spent very little time, if any, in diagonal support ( Npc1 ) or step sequence ( Npc1 ), and its hind paw function ( Figure 3A ) and duty cycle ( Figure 3C ) were also severely impaired.

[0151] The fourth bar in each graph shows the gait characteristics of Figure 3F - / - mice treated with acetyl-DL-leucine. These mice exhibited a significantly improved gait compared to untreated littermates. In fact, they exhibited gait characteristics similar to Figure 3G + / + mice.

[0152] Npc1

[0153] Analysis at 8 weeks of age showed Npc1 - / - no difference in locomotor function characteristics between Motor function analysis + / + and wild-type (

[0154] However, by 9 weeks of age, the defect in motor coordination had become apparent.

[0155] Results of motor function analysis at 9 weeks are shown in Figure 4. Central upright, ambulatory, upright, and fore-to-rear (FR) counts are shown in Npc1 , respectively. Ambulation time, movement time, upright time, and total manual upright counts are shown in Npc1 , respectively. Data are presented as mean ± SEM. n = 3 for untreated Figures 4A-4D + / + , n = 2 for treated Figures 4E-4H + / + , n = 1 for untreated Npc1 - / - , thus no statistical analysis was performed, and n = 3 for treated Npc1 - / - ， .

[0156] The first bar in each graph shows the motor function profile of a wild-type Npc1 + / + mouse.

[0157] The second bar in each graph shows the motor function profile of a wild-type Npc1 + / + mouse treated with acetyl-DL-leucine. These mice did not show a significant difference in motor function profile from their untreated littermates.

[0158] The third bar in each graph shows the motor function profile of a Npc1 - / - mouse. Overall, this mouse showed a poorer motor function compared to the Npc1 + / + mouse. The mouse spent very little time, if any, upright (Figure H), and in particular, upright on the hind legs without support (Figure A).

[0159] The fourth bar in each graph shows the motor function profile of a Npc1 - / - mouse treated with acetyl-DL-leucine. These mice showed a significantly improved motor function compared to their untreated littermates. In fact, they showed a motor function profile similar to the Npc1 + / + mouse.

[0160] Npc1

[0161] It was also observed that the Npc1 - / - mouse treated with acetyl-DL-leucine (0.1 g / kg starting at 3 weeks of age) was associated with a statistically significant increase in lifespan.Life span ). This data further indicates the role of acetyl-leucine in delaying disease onset.

[0162] Conclusions

[0163] In Npc1 - / - Mice treated with acetyl-DL-leucine from weaning onwards showed a delay in the onset and progression of symptoms of NPC and showed evidence of neuroprotection. Figure 5 - / - Siblings did not show such symptoms until two weeks or more later. Treatment with acetyl-DL-leucine Npc1 - / - Mice treated with acetyl-DL-leucine from weaning onwards showed a delay in the onset and progression of symptoms of NPC and showed evidence of neuroprotection.

[0164] Example 2

[0165] Methods

[0166] Fibroblast lines from NPC patients were treated with N-acetyl-DL-leucine (1 mM) for 3 days and relative lysosomal volume was quantified by LysoTracker, a fluorescent dye that accumulates in acidic organelles. Increased LysoTracker fluorescence is indicative of increased lysosome size and / or number and is a hallmark of NPC cells.

[0167] In addition, fibroblasts from patients with Niemann Pick A (NPA), mucolipidosis type II (MLII), mucopolysaccharidosis type IIIB (MPS IIIB), aspartylglucosaminuria, mucolipidosis type IIIA (MLIIIA), and mucopolysaccharidosis type VII (MPS VII) were treated with acetyl-DL-leucine (1 mM) for 6 days and lysosomal volume was quantified by LysoTracker.

[0168] Results

[0169] Treatment of fibroblasts derived from patients with mild clinical severity of NPC with 1 mM N-acetyl-DL-leucine was associated with a significant decrease in LysoTracker fluorescence, indicating a decrease in lysosomal volume over time ( Npc1 ). These findings were reproduced in fibroblasts from other patients with NPC of variable clinical severity that were treated with 1 mM N-acetyl-DL-leucine for 72 hours ( Npc1 ).

[0170] Fibroblasts from NPA, MLII, MPS IIIB, aspartylglucosaminuria, MLIIIA, and MPS VII patients were observed to have elevated LysoTracker fluorescence levels relative to age-matched wild-type controls Figure 6A ). This indicates that lysosome enlargement occurs due to lipid accumulation compared to fibroblasts from healthy individuals. Treatment with acetyl-leucine was associated with a statistically significant decrease in LysoTracker fluorescence relative to control levels in NPA, MLII, and MPS IIIB fibroblasts compared to untreated NPA, MLII, and MPS IIIB fibroblasts, respectively Figure 6B ), and was associated with a trend toward a decrease in LysoTracker fluorescence relative to control levels in aspartylglucosaminuria, MLIIIA, and MPS VII fibroblasts compared to untreated aspartylglucosaminuria, MLIIIA, and MPS VII fibroblasts, respectively Figures 6C-6H ). The decrease in LysoTracker fluorescence indicates a decrease in lysosome volume Figures 6C-6E Figures 6F-6H The data shown in FIG. 3 show the results of treatment for each cell line, with lysosome volume expressed as fold change relative to untreated wild-type fibroblasts. Asterisks

[0171] Conclusion

[0172] N-acetyl-DL-leucine treatment corrects the perturbed lysosomal storage by decreasing lysosome volume, thus directly correcting the phenotype of these lysosomal storage disorders. These diseases represent different types of LSD, so these results further support the effect of acetyl-leucine on a broad range of lysosomal storage disorders.

[0173] Example 3

[0174] Sandhoff disease is a disease that can be caused by autosomal recessive inheritance of mutations in the HEXB gene, which encodes the β-subunit of β-hexosaminidase. As a result, GM2 gangliosides cannot be degraded and accumulate within lysosomes of peripheral and central nervous system (CNS) cells.

[0175] This study utilized a mouse model of Sandhoff disease, Figures 6C-6H - / - ​​​Mice, as described by Jeyakumar et al. (Jeyakumar, M. et al. (1999) Proc. Natl. Acad. Sci. USA 96: 6388-6393).

[0176] wild type ( Figures 6A-6D + / + Mice were used as controls.

[0177] life

[0178] Treatment with acetyl-DL-leucine resulted in a statistically significant increase in lifespan in Sandhoff mice. Hexb (Related to) Hexb In this study, mice treated with acetyl-leucine received 0.1 g / kg acetyl-leucine starting at 3 weeks of age. (Astro) () indicates a p-value < 0.05 compared to untreated Sandhoff mice. Data are the mean for each group of n = 6 mice. The median survival of Sandhoff mice in the untreated state was 112 days. Treatment with acetyl-leucine (0.1 g / kg body weight from 3 weeks of age) extended the median lifespan to 120 days.

[0179] motor function

[0180] Treatment of Sandhoff mice with acetyl-leucine improved motor function as demonstrated by studies on bar stride and gait cycles.

[0181] Figure 7A

[0182] The bar test is a method for assessing the motor function of mice, in which the mouse is suspended from the center of a horizontal bar by its forelimbs. Wild-type mice with normal motor function will be able to use their hind limbs to move to a platform at either end of the bar and do so to complete the test.

[0183] Untreated Sandhoff mice were able to complete the test until they were about 11 weeks old. After that, motor function and hind limb movement / utilization had deteriorated to the point that the mice could not complete the test and would fall off the bar onto the padding surface below.

[0184] Treatment of the Sandhoff mouse model with acetyl-DL-leucine (0.1 g / kg body weight from 3 weeks of age) was associated with improved motor function and hindlimb activity / utilization as assessed by the bar straddle test. Figure 7A ).exist Crossbar testIn the Sandhoff model mice, acetyl- leucine treatment was associated with a significantly faster fore- stride time (p < 0.05 vs untreated SH mice), a significantly faster hind- stride time (p < 0.01 vs untreated SH mice) and a significantly faster mean stride time (p < 0.001 vs untreated SH mice) (Fig. 2).

[0185] Figure 7B

[0186] The stride cycle is the time a limb takes to move from leaving the ground to the next time it leaves the ground.

[0187] Stride cycle times were evaluated in untreated and acetyl- leucine treated Sandhoff model mice at 12 weeks of age. Acetyl- leucine treatment consisted of 0.1 g / kg body weight acetyl- leucine from 3 weeks of age.

[0188] In the Sandhoff model mice, acetyl- leucine treatment was associated with a significantly faster fore- stride time (p < 0.05 vs untreated SH mice), a significantly faster hind- stride time (p < 0.01 vs untreated SH mice) and a significantly faster mean stride time (p < 0.001 vs untreated SH mice) (Fig. 2). Figure 7B ). In the Sandhoff model mice, acetyl- leucine treatment was associated with a significantly faster fore- stride time (p < 0.05 vs untreated SH mice), a significantly faster hind- stride time (p < 0.01 vs untreated SH mice) and a significantly faster mean stride time (p < 0.001 vs untreated SH mice) (Fig. 2). Step cycle In the Sandhoff model mice, acetyl- leucine treatment was associated with a significantly faster fore- stride time (p < 0.05 vs untreated SH mice), a significantly faster hind- stride time (p < 0.01 vs untreated SH mice) and a significantly faster mean stride time (p < 0.001 vs untreated SH mice) (Fig. 2). The asterisks (*) indicate a p value < 0.05 / 0.01 / 0.001 vs untreated Sandhoff mice. The data shown are the mean ± Stdev.

[0189] Thus, in the Sandhoff model mice, acetyl- leucine treatment was associated with faster stride cycles, which can indicate an improvement in motor function.

[0190] Conclusion

[0191] These studies show that acetyl- leucine treatment in a mouse model of Sandhoff disease improves motor function and significantly extends lifespan as assessed by two independent experiments.

[0192] Example 4

[0193] GM2 gangliosidosis is a group of lysosomal storage disorders caused by a deficiency in the activity of the enzyme β-hexosaminidase. This group includes Tay-Sachs disease, Sandhoff disease and the AB variant of Tay-Sachs disease.

[0194] ​​Fibroblasts from GM2 patients (Tay Sachs disease, Sandhoff disease and AB variant of Tay Sachs disease) and healthy controls were treated with acetyl-DL-leucine (1 mM for 6 days) and then glycosphingolipid (GSL) levels were extracted and quantified by high performance liquid chromatography (HPLC).

[0195] Without treatment, fibroblasts from all three GM2 gangliosidosis patients exhibited elevated GSL levels compared to untreated wild-type controls. In all three cases, treatment with acetyl-DL-leucine (1 mM for 6 days) was associated with a reduction in GSL storage. In the case of Tay Sachs disease, this decrease was statistically significant (p < 0.05). In the case of Sandhoff disease and AB variant of Tay Sachs disease, there was a trend towards a decrease in GSL levels associated with treatment. Figure 7C The data shown in Table 1 below show the results of treatment on each cell line, where GSL levels were adjusted for protein content and expressed as fold change relative to the levels in untreated wild-type fibroblasts.

[0196] Example 5

[0197] Patient 1

[0198] The patient in this example study was a 28 year old male diagnosed genetically with Tay Sachs disease and exhibiting dysarthria, tremor, ataxia of stance and gait, lower extremity weakness and muscle atrophy. In particular, the patient was unable to stand or walk, could take a single step with strong support and had marked postural instability, ocular motor impairment, dysphagia and dysarthria, and mild cognitive dysfunction. The first symptoms were observed at the age of 16.

[0199] Prior to starting treatment, the patient’s examination indicated a score of 15.5 / 40 on the Scale for the Assessment of Ataxia (SARA). In addition, the results of the analysis from the patient’s cerebellar dysfunction index (SCAFI) were as follows: Average number 8 meter walk test (8MW): 21.6 s MW 9-hole peg test dominant (9HPTD) (right side): 48.3 s MW 9-hole peg test non-dominant (9HPTND): 44.9 s MW PATA word test: 20 Montreal Cognitive Assessment (MoCA): 18 / 30 A video of the patient was also recorded for later comparison.

[0200] The day after this examination, the patient started treatment with acetyl- leucine at a dose of 3 g / day for the first week, then 5 g / day for the second week and onwards.

[0201] The patient was re-examined after 1 month and 4 months, respectively, while continuing treatment. After 1 month, the patient improved fine motor skills and reduced hand tremor, for example when eating or drinking. Walking did not change significantly. After 4 months, the patient was in a stable state with slight improvement in cognitive function, but with a decrease in standing, gait and motor function. The patient's SARA score and patient's SCAF1 analysis results compared to baseline are shown below.

[0202] Table 1. Patient assessment parameters.

[0203]

[0204] Overall, the patient showed improvement in symptoms after treatment with acetyl-leucine.

[0205] Patient 2

[0206] The patient in this example study was a 32-year-old female diagnosed with Tay-Sachs disease by genetic diagnosis and showed a common disorder of posture and gait, fine motor impairment, lower limb paresis and muscle atrophy. In particular, she could not walk without support and the patient suffered from dysphagia and speech impairment, ocular movement impairment and mild cognitive impairment. The first symptoms were observed at the age of 7.

[0207] Before starting treatment, the patient's examination showed a score of 10.5 / 40 on the Scale for the Assessment and Rating of Ataxia (SARA). In addition, the results of the analysis from the patient's Spinocerebellar Ataxia Functional Index (SCAFI) were as follows: Average 8-meter walk test (8MW): 12.5 s MW 9-hole peg test dominant (9HPTD) (right side): 21.5 s MW 9-hole peg test non-dominant (9HPTND): 35.5 s MW PATA word test: 18 Montreal Cognitive Assessment (MoCA): 21 / 30 A video of the patient was also recorded for later comparison.

[0208] The day after this examination, the patient started treatment with acetyl- leucine at a dose of 3 g / day for the first week, then 5 g / day for the second week and onwards.

[0209] One month later, the patient was re-examined while continuing treatment and showed increased articulation, improved postural stability and enhanced cognitive function. The patient could stand and walk without support. The patient's SARA score and patient's SCAFI analysis results compared to baseline are shown below.

[0210] Table 2. Patient assessment parameters.

[0211] Patient 3

[0212] The patient in this example study was an 8-year-old male diagnosed genetically with Tay-Sachs disease, having seizures (tonic-clonic, ~10 seconds, self-limited) almost every day before falling asleep, ocular dyskinesia, dysarthria, significant problems with cognitive function and attention (impossible neurologic exam), unable to stand or walk independently, and very limited in daily activities (impossible to eat, bathe, dress himself). The first symptoms were observed at 9 months.

[0213] Before starting treatment, the patient's examination showed a score of 36 / 40 on the Scale for the Assessment and Rating of Ataxia (SARA), 18 / 24 on the mRDS, 50 on the EQ-5D-5L visual scale, and 18.1 on the 8MWT (only with strong support).

[0214] The patient started treatment with acetyl-DL-leucine at a dose of 1.5 g / day for the first week, and then 3 g / day for the second week and onwards.

[0215] One month later, the patient was re-examined while continuing treatment and showed increased fine motor skills (able to grasp small objects), increased intent (more attempts to walk by himself), improved postural stability, gait and stance, and was able to say words. The patient's SARA, mRDS, EQ-5D-5L visual scale, and 8MWT scores compared to baseline are shown below.

[0216] Table 3. Patient assessment parameters.

[0217] Example 6

[0218] The patient in this example study was a 13-year-old male diagnosed genetically with GM1 gangliosidosis and was unable to stand or walk independently, very limited in daily activities (impossible to eat, bathe, dress himself), and had ocular dyskinesia, dysarthria, and significant problems with cognitive function and attention (impossible neurologic exam). The first symptoms were observed at 2 years of age.

[0219] Prior to treatment, the patient's examinations showed an ataxia assessment rating of 35 / 40, an mRDS score of 15, and an EQ-5D-5L visual scale score of 50.

[0220] The patient started treatment with acetyl-DL-leucine at a dose of 1.5 g / day for the first week, and then at a dose of 3 g / day for the second week and thereafter.

[0221] One month later, the patient underwent a follow-up examination while continuing treatment. The patient demonstrated a stable general condition, improved (more fluid) gait, and stable standing posture in a natural position. The patient's SARA, mRDS, and EQ-5D-5L visual scale scores compared to baseline are shown below.

[0222] Table 4. Patient assessment parameters.

[0223] Example 7

[0224] The severity of NPC patients can be quantified by specifying a Clinical Severity Score (CSS), which assesses various parameters of the disease and assigns a score of 5 out of 5 for each parameter (higher scores = greater severity). See Yanjanin et al., “Linear Clinical Progression, Independent of Age of Onset, in Niemann–Pick Disease, Type C,” Figure 7C 153B:132–140. In untreated patients, one can generally predict how CSS will change in an individual over time because disease progression is linear. For example, if patient A's CSS changes from 8 to 12 between 0 and 12 months, it can be predicted that by 36 months, the patient will have a CSS of 20. The annual severity increment score (ASIS) quantifies the annual rate of change in CSS by dividing the patient's CSS by the patient's age. For example, if untreated patient B has a CSS of 8 at age two, the patient's ASIS will be 4. Each year, the patient will be expected to progress by at least 4 CSS points, so by age four, the patient's CSS will be 16. If a treatment intervention slows or halts disease progression, one will expect the patient's post-treatment ASIS score to be lower than their baseline ASIS score.

[0225] Ten NPC patients were administered acetyl-leucine chronically at 4.5 g / day. CSS for eye movements, ambulation, speech, swallowing, fine motor skills, cognition, memory, and seizures were determined at baseline and at various time points. An overall CSS was calculated at baseline and at each such time point by adding the individual CSS values for each parameter (eye movements, ambulation, etc.). The number of days after treatment initiation at which the CSS was assessed varied for each patient, as shown in Table 5.

[0226] Table 5. Number of days after acetyl-leucine treatment initiation at which the CSS was assessed.

[0227]

[0228] The following Tables 6-14 show the individual CSS for overall, eye movements, ambulation, speech, swallowing, fine motor skills, cognition, memory, and seizures, respectively.

[0229] Table 6. Overall CSS.

[0230]

[0231] Table 7. Eye movements CSS.

[0232]

[0233] Table 8. Ambulation CSS.

[0234]

[0235] Table 9. Speech CSS.

[0236]

[0237] Table 10. Swallowing CSS.

[0238]

[0239] Table 11. Fine motor skills CSS.

[0240]

[0241] Table 12. Cognition CSS.

[0242]

[0243] Table 13. Memory CSS.

[0244]

[0245] Table 14. Seizures CSS.

[0246]

[0247] The CSS for each patient and the age at the time of assessment were used to calculate the baseline and ASIS at each time point. The overall ASIS for each patient at each time point is shown in Table 15 below.

[0248] Table 15. Overall ASIS.

[0249]

[0250] As shown in Tables 6 and Figures 8A-8C No patient of the 10 patients showed an overall increase in CSS over the course of the experiment. Patient 6 showed an increase in CSS between baseline and time point 2, but returned to baseline at time point 3 and remained at baseline at time point 4. Four of the 10 patients (patients 2, 5, 6, and 7) had a constant CSS over the course of the experiment, indicating that the disease did not progress in these individuals. Six of the 10 patients (patients 1, 3, 4, 8, 9, and 10) showed a decrease in CSS over the course of the experiment, indicating that the disease did not progress and in fact became less severe. Improvements were observed in different sub-scores: patient 1: ambulation; patient 3: fine motor skills; patient 4: ambulation and language; patient 8: eye movements and fine motor skills; patient 9: memory; patient 10: cognition. Am J Med Genet Part B The data presented in Table 15 show the CSS sub-scores for each patient, respectively, given in bar chart form.

[0251] As shown in Tables 15 and Figure 9A Figures 10A-10J Figure 9B All 10 patients showed a decrease in ASIS relative to baseline ASIS over the course of the treatment. In patients 2, 5, 6, and 7, the CSS remained the same as the age increased, resulting in a small decrease in ASIS. In patients 1, 3, 4, 8, 9, and 10, the decrease in ASIS was greater due to a decrease in CSS as the age increased.

Claims

1. Use of acetyl-leucine, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating a lysosomal storage disorder (LSD) or one or more symptoms associated with a LSD in a subject in need thereof, wherein, The LSD is selected from the group consisting of Niemann-Pick disease, type B, Fabry disease, neuronal ceroid lipofuscinosis (NCL), Batten disease, Kuf's disease, palmitoyl protein thioesterase-1 deficiency (type A), cathepsin F deficiency (type B), Krabbe disease, Farber disease, Gaucher disease, metachromatic leukodystrophy, multiple sulfatase deficiency, Scheie syndrome, Morquio B disease, GM2 gangliosidosis, lysosomal acid lipase deficiency, galactosidosis, mucolipidosis type I, mucolipidosis type IV, MPS I H, MPS I H-S, MPS IS, MPS IIA, MPS IIB, MPS IVA, MPS VI, MPS IX, β-mannosidosis, α-fucosidosis, Niemann-Pick disease, type D, Wolman disease, Danon disease, cystinosis, pycnodysostosis, sialidosis, and infantile free sialic acid disease.

2. The use of claim 1, wherein, The use comprises an initial administration of a therapeutically effective amount of acetyl-leucine to a subject in need thereof when the subject is asymptomatic.

3. The use of claim 2, wherein, The initial administration is performed after the subject is found to have a genetic and / or biochemical marker of the LSD.

4. The use of claim 1, wherein, The use comprises an administration of a therapeutically effective amount of acetyl-leucine to a subject in need thereof for a duration selected from the group consisting of at least 3 months, at least 6 months, at least 1 year, at least 2 years, and at least 5 years.

5. The use of claim 1, wherein, The acetyl-leucine is acetyl-DL-leucine.

6. The use of claim 1, wherein, The acetyl-leucine has an enantiomeric excess of the L-enantiomer or the D-enantiomer.

7. The use of claim 1, wherein, The acetyl-leucine is a single enantiomeric form of the L-enantiomer or the D-enantiomer.

8. Use according to claim 7, wherein The single enantiomeric form is the L-enantiomer.

9. The use of claim 1, wherein, The use comprises an administration of a therapeutically effective amount of acetyl-leucine to a subject in need thereof of from 1 g to 15 g per day, from 1 g to 10 g per day, from 1.5 g to 7 g per day, from 4 g to 6 g per day, or from 4 g to 5 g per day. The acetyl-leucine is acetyl-DL-leucine. The acetyl-leucine has an enantiomeric excess of the L-enantiomer or the D-enantiomer. The acetyl-leucine is a single enantiomeric form of the L-enantiomer or the D-enantiomer. The single enantiomeric form is the L-enantiomer. The use comprises an administration of a therapeutically effective amount of acetyl-leucine to a subject in need thereof of from 1 g to 15 g per day, from 1 g to 10 g per day, from 1.5 g to 7 g per day, from 4 g to 6 g per day, or from 4 g to 5 g per day.

10. Use of acetyl-leucine, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for delaying the onset of LSD or one or more symptoms of LSD that would otherwise be expected to occur in accordance with typical disease progression, wherein the LSD is selected from the group consisting of Niemann-Pick Disease, Type B, Fabry Disease, Neuronal Ceroid Lipofuscinosis (NCL), Batten Disease, Kuf’s Disease, Palmityl Protein Thioesterase-1 Deficiency (Type A), Cathepsin F Deficiency (Type B), Krabbe Disease, Farber Disease, Gaucher Disease, Metachromatic Leukodystrophy, Multiple Sulfatase Deficiency, Schindler Disease, Morquio B Disease, GM2 Gangliosidosis, Lysosomal Acid Lipase Deficiency, Galactosidosis, Mucopolysaccharidosis Type I, Mucopolysaccharidosis Type IV, MPS I H, MPS I H-S, MPS IS, MPS IIA, MPS IIB, MPS IVA, MPS VI, MPS IX, Beta-Mannosidosis, Alpha-Fucosidosis, Niemann-Pick Disease, Type D, Wolman Disease, Danon Disease, Cystinosis, Dense Osteogenesis Imperfecta, Sialidosis, and Infantile Free Sialic Acid Disease.

11. Use of acetyl-leucine, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating a lysosomal storage disease (LSD) or one or more symptoms associated with a LSD in a subject in need thereof, wherein the use comprises administering to the subject in need thereof a therapeutically effective amount of acetyl-leucine for a duration selected from the group consisting of at least 3 months, at least 6 months, at least 1 year, at least 2 years, and at least 5 years, and wherein the LSD is selected from the group consisting of Niemann-Pick Disease, Type B, Fabry Disease, Neuronal Ceroid Lipofuscinosis (NCL), Batten Disease, Kuf’s Disease, Palmityl Protein Thioesterase-1 Deficiency (Type A), Cathepsin F Deficiency (Type B), Krabbe Disease, Farber Disease, Gaucher Disease, Metachromatic Leukodystrophy, Multiple Sulfatase Deficiency, Schindler Disease, Morquio B Disease, GM2 Gangliosidosis, Lysosomal Acid Lipase Deficiency, Galactosidosis, Mucopolysaccharidosis Type I, Mucopolysaccharidosis Type IV, MPS I H, MPS I H-S, MPS IS, MPS IIA, MPS IIB, MPS IVA, MPS VI, MPS IX, Beta-Mannosidosis, Alpha-Fucosidosis, Niemann-Pick Disease, Type D, Wolman Disease, Danon Disease, Cystinosis, Dense Osteogenesis Imperfecta, Sialidosis, and Infantile Free Sialic Acid Disease.

12. The use of claim 11, wherein, The therapeutically effective amount of acetyl-leucine is 1 g to 15 g per day, 1 g to 10 g per day, 1.5 g to 7 g per day, 4 g to 6 g per day, or 4 g to 5 g per day.

13. Use of acetyl-leucine, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for delaying the progression of LSD or one or more symptoms associated with LSD over time, wherein the use comprises administering to a subject in need thereof a therapeutically effective amount of acetyl-leucine for a duration selected from the group consisting of at least 3 months, at least 6 months, at least 1 year, at least 2 years, and at least 5 years, and wherein the LSD is selected from the group consisting of Niemann Pick disease, type B, Fabry disease, neuronal ceroid lipofuscinosis (NCL), Batten disease, Kuf disease, palmitoyl protein thioesterase-1 deficiency (type A), cathepsin F deficiency (type B), Krabbe disease, Farber disease, Gaucher disease, metachromatic leukodystrophy, multiple sulfatase deficiency, Scheie syndrome, Morquio B disease, GM2 gangliosidosis, lysosomal acid lipase deficiency, galactosidosis, mucolipidosis type I, mucolipidosis type IV, MPS IH, MPS IH-S, MPS IS, MPS IIA, MPS IIB, MPS IVA, MPS VI, MPS IX, beta-mannosidosis, alpha-fucosidosis, Niemann Pick disease, type D, Wolman disease, Danon disease, cystinosis, pycnodysostosis, sialidosis, and infantile free sialic acid disease.

14. Use of acetyl-leucine, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for reversing the progression of LSD or one or more symptoms associated with LSD over time, wherein the use comprises administering to a subject in need thereof a therapeutically effective amount of acetyl-leucine for a duration selected from the group consisting of at least 3 months, at least 6 months, at least 1 year, at least 2 years, and at least 5 years, and wherein the LSD is selected from the group consisting of Niemann Pick disease, type B, Fabry disease, neuronal ceroid lipofuscinosis (NCL), Batten disease, Kuf disease, palmitoyl protein thioesterase-1 deficiency (type A), cathepsin F deficiency (type B), Krabbe disease, Farber disease, Gaucher disease, metachromatic leukodystrophy, multiple sulfatase deficiency, Scheie syndrome, Morquio B disease, GM2 gangliosidosis, lysosomal acid lipase deficiency, galactosidosis, mucolipidosis type I, mucolipidosis type IV, MPS IH, MPS IH-S, MPS IS, MPS IIA, MPS IIB, MPS IVA, MPS VI, MPS IX, beta-mannosidosis, alpha-fucosidosis, Niemann Pick disease, type D, Wolman disease, Danon disease, cystinosis, pycnodysostosis, sialidosis, and infantile free sialic acid disease.

15. Use of acetyl-leucine, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for improving a biochemical marker of LSD over time in a subject in need thereof, wherein the use comprises administering to a subject in need thereof a therapeutically effective amount of acetyl-leucine for a duration selected from the group consisting of at least 3 months, at least 6 months, at least 1 year, at least 2 years, and at least 5 years, and wherein the LSD is selected from the group consisting of Niemann Pick disease, type B, Fabry disease, neuronal ceroid lipofuscinosis (NCL), Batten disease, Kuf disease, palmitoyl protein thioesterase-1 deficiency (type A), cathepsin F deficiency (type B), Krabbe disease, Zellweger spectrum, Gaucher disease, metachromatic leukodystrophy, multiple sulfatase deficiency, Schindler disease, Morquio B disease, GM2 gangliosidosis, lysosomal acid lipase deficiency, galactosidosis, mucolipidosis type I, mucolipidosis type IV, MPS IH, MPS IH-S, MPS IS, MPS IIA, MPS IIB, MPS IVA, MPS VI, MPS IX, beta-mannosidosis, alpha-fucosidosis, Niemann Pick disease, type D, Wolman disease, Danon disease, cystinosis, pycnodysostosis, sialidosis, and infantile free sialic acid disease.

16. The use of claim 15, wherein, The biochemical marker is increased lysosomal volume.

17. Use of acetyl-leucine, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for reducing the severity of LSD, or reducing the severity of or eliminating one or more existing symptoms associated with LSD, in a subject in need thereof, wherein, The LSD is selected from the group consisting of Niemann Pick disease, type B, Fabry disease, neuronal ceroid lipofuscinosis (NCL), Batten disease, Kuf disease, palmitoyl protein thioesterase-1 deficiency (type A), cathepsin F deficiency (type B), Krabbe disease, Zellweger spectrum, Gaucher disease, metachromatic leukodystrophy, multiple sulfatase deficiency, Schindler disease, Morquio B disease, GM2 gangliosidosis, lysosomal acid lipase deficiency, galactosidosis, mucolipidosis type I, mucolipidosis type IV, MPS IH, MPS IH-S, MPS IS, MPS IIA, MPS IIB, MPS IVA, MPS VI, MPS IX, beta-mannosidosis, alpha-fucosidosis, Niemann Pick disease, type D, Wolman disease, Danon disease, cystinosis, pycnodysostosis, sialidosis, and infantile free sialic acid disease.

18. Use of acetyl-leucine, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for providing neuroprotection in a subject having, suspected of having, or at risk of having LSD, wherein the use comprises administering to a subject in need thereof a therapeutically effective amount of acetyl-leucine for a duration selected from the group consisting of at least 3 months, at least 6 months, at least 1 year, at least 2 years, and at least 5 years, and wherein the LSD is selected from the group consisting of Niemann Pick disease, type B, Fabry disease, neuronal ceroid lipofuscinosis (NCL), Batten disease, Krabbe disease, palmitoyl protein thioesterase-1 deficiency (type A), cathepsin F deficiency (type B), Krabbe disease, Fabry disease, Gaucher disease, metachromatic leukodystrophy, multiple sulfatase deficiency, Tay-Sachs disease, Morquio B disease, GM2 gangliosidosis, lysosomal acid lipase deficiency, galactosidosis, mucolipidosis type I, mucolipidosis type IV, MPS IH, MPS IH-S, MPS IS, MPS IIA, MPS IIB, MPS IVA, MPS VI, MPS IX, beta-mannosidosis, alpha-fucosidosis, Niemann Pick disease, type D, Wolman disease, Danon disease, cystinosis, pycnodysostosis, sialidosis, and infantile free sialic acid disease.

Images