Pharmaceutical compositions and uses against lysosomal storage disorders

Abstract

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

Description

[0001] This application is a divisional application. The international application number of the original application is PCT / IB2017 / 054928, the application date is August 11, 2017, the Chinese national phase application number is 201780062740.X, and the invention title is "Pharmaceutical Composition and Use for Lysosomal Storage Disorders".

[0002] This application claims priority to 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 these documents 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 more than 70 diseases with a clinical frequency of 1 in 5000 live births. These diseases can be divided into two main categories: primary storage disorders caused by a direct deficiency of degradation pathways (usually lysosomal enzyme deficiencies), and secondary storage disorders caused by dysfunction of downstream lysosomal proteins or processes affecting lysosomes (e.g., defects in transport pathways).

[0004] The pathology of LSD 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 disease is typically severe and uninterrupted. It tends to appear in the first few years of life, and severe progression leads to frequent hospitalizations. If left untreated, patients often die in their teens. Adult-onset patients have also been described.

[0005] Currently, treatment options for LSD are limited. Treatment options are scarce, and many, if available, only improve quality of life. For example, some LSD cases respond to bone marrow transplantation or enzyme replacement therapy. Furthermore, some benefits have been reported in clinical trials using substrate reduction therapy (SRT) with the imino sugar drug Miglustastat, a glycosphingolipid (GSL) biosynthesis inhibitor (Patterson, 2006). However, there is currently no universal, nonspecific treatment beneficial for all LSD cases. Therefore, there is a need to develop improved treatments for LSD.

[0006] This disclosure addresses this need and describes acetyl-leucine for treating LSD or one or more symptoms of LSD in subjects in need.

[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, wherein the LSD is not Niemann-Pick type C disease.

[0008] In one embodiment of this disclosure, acetyl-leucine or a pharmaceutically acceptable salt thereof is disclosed for use in a method of treating LSD in a subject who 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, the disease or symptoms of which would otherwise be expected to occur according to typical disease progression.

[0010] In yet another embodiment, this disclosure includes acetyl-leucine or a pharmaceutically acceptable salt thereof, used in a method for treating LSD or one or more symptoms associated with LSD in a subject in need, wherein the method includes administering a therapeutically effective amount of acetyl-leucine to the subject in need for a duration selected from 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, this 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 LSD-related symptoms over time compared with typical disease progression, wherein the method includes administering a therapeutically effective amount of acetyl-leucine to a subject in need for a duration selected from 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 LSD-related symptoms over time, wherein the method comprises administering a therapeutically effective amount of acetyl-leucine to a subject in need for a duration selected from 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 biochemical markers of LSD in a subject of need over time, wherein the method comprises administering a therapeutically effective amount of acetyl-leucine to the subject of need for a duration selected from 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, this disclosure includes acetyl-leucine or a pharmaceutically acceptable salt thereof, used in methods for reducing the severity of LSD in subjects in need, or for reducing the severity of one or more existing symptoms associated with LSD, or for eliminating one or more existing symptoms associated with LSD, wherein the LSD is not Niemann-Pick type C disease.

[0015] In yet another embodiment, this disclosure includes acetyl-leucine or a pharmaceutically acceptable salt thereof, used in a method of improving neuroprotection in subjects who have, are suspected of having, or are at risk of having LSD, wherein the method includes administering a therapeutically effective amount of acetyl-leucine to the subject for a duration selected from 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 this disclosure include the use of acetyl-leucine or a pharmaceutically acceptable salt thereof in a method for delaying the progression of lysosomal storage disease (LSD) in a subject. Acetyl-leucine or a pharmaceutically acceptable salt thereof is also used in a method for providing neuroprotection to a subject with LSD. Acetyl-leucine is in a racemic form, with an enantiomeric excess of the L-enantiomer or the D-enantiomer. The method further includes administration of acetyl-leucine at a dose of 1.5 g to 10 g / day. Furthermore, the method includes administration of acetyl-leucine for treatment lasting more than two weeks. The method may also include administration of acetyl-leucine or a pharmaceutically acceptable salt thereof prior to the onset of LSD symptoms. The method may also include administration of another therapy or agent for the prevention or treatment of LSD. A further embodiment of this disclosure is a kit for delaying the progression of LSD in a subject, the kit comprising: a device for diagnosing or predicting LSD, and acetyl-leucine or a pharmaceutically acceptable salt thereof. The kit comprises: a device for diagnosing or predicting LSD, and acetyl-leucine or a pharmaceutically acceptable salt thereof. Another embodiment of this disclosure is the use of acetyl-leucine or a pharmaceutically acceptable salt thereof as a neuroprotective agent in subjects with LSD. In further embodiments of the methods, kits, or applications, LSD is Niemann-Pick disease type C (NPC1 and / or NPC2 deficiency), Smith-Lemli-Opitz Syndrome (SLOS), congenital errors in cholesterol synthesis, Tangier disease, Pelizaeus-Merzbacher disease, neuronal ceroid lipofuscin deposition, primary glycosphingolipidosis, Farber disease, or multiple sulfatase deficiencies. In addition, in another embodiment of the method, kit, or application, the primary glycosphingolipid disease is Gaucher disease, Fabry disease, GM1 gangliosidosis, GM2 gangliosidosis, Krabbe disease, or metachromatic leukodystrophy (MLD).In a further embodiment of the method, kit, or application, LSD is NPC, Tay-Sachs disease, Sandhoff disease, GM1 ganglioside storage disease, Fabry disease, neurodegenerative mucopolysaccharidosis, MPS I, MPS IH, MPS IS, MPS II, MPS III, MPS IIIA, MPSIIIB, 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 Tangier disease. In another embodiment of the method, kit, or application, LSD is Niemann-Pick disease, Niemann-Pick disease type C, Niemann-Pick disease type A, Sandhoff disease, Tay-Sachs disease, or mucopolysaccharidosis type II.

[0017] These and other embodiments and features of this disclosure will become apparent from the following description and claims. Attached Figure Description

[0018] Figure 1 shows the treated ( Figure 1A ) and untreated ( Figure 1B 9 weeks old Npc1 - / - A photo of a mouse.

[0019] Figure 2A and 2B The results showed that, with and without acetyl-DL-leucine treatment from weaning, Npc1 - / - Mice and wild-type ( Npc1 + / + Weight data compared to mice.

[0020] Figure 3A-3G The results showed that, with and without acetyl-DL-leucine treatment from weaning, Npc1 - / - Mice and wild-type ( Npc1 + / + Gait analysis data compared to mice. For example, diagonal support, gait frequency, and gait sequence data are shown below. Figures 3A-3C As shown. Figure 3D and 3E The forepaw (FP) data are shown (standing mean and stride cycle in mosaic D; duty cycle in mosaic E). Figure 3F and 3GThe hind paw (HP) data are shown (standing average and stride cycle in mosaic F; duty cycle in mosaic G).

[0021] Figures 4A-4H The results showed that, with and without acetyl-DL-leucine treatment from weaning, Npc1 - / - Mice and wild-type ( Npc1 + / + Motor function analysis data compared to mice. Centered feeding, activity, feeding, and front-to-back (FR) counts are shown in the figures. Figures 4A-4D The activity time, movement time, feeding time, and total manual feeding count are as follows: Figure 4E-4H As shown.

[0022] Figure 5 Treatment with acetyl-DL-leucine (0.1 g / kg from 3 weeks of age) was shown to be associated with a small but statistically significant increase in lifespan in Npc1- / - mice.

[0023] Figure 6A and 6B The study showed a decrease in lysosomal volume in non-neuronal NPC cells after treatment with acetyl-DL-leucine. Figure 6C-6H The effects of acetyl-DL-leucine treatment on lysosomal volume in fibroblasts from patients with NPA, ML II, MPS IIIB, aspartic glucosamineuria, MLIIIA, and MPS VII were shown.

[0024] Figure 7A Survival curves representing mortality rates in untreated or acetyl-leucine-treated wild-type and Sandhoff mice are shown. Figure 7B Displays bar crossing scores in untreated and acetyl-leucine-treated Sandhoff model mice. Figure 7C The gait cycle time of untreated and acetyl-leucine-treated Sandhoff mice at 12 weeks of age is shown.

[0025] Figures 8A-8C This study demonstrates the effect of acetyl-DL-leucine treatment on glycosphingolipid (GSL) levels in fibroblasts of patients with GM2 ganglioside storage disorders (Tysachus disease, Sandhoff disease, and the AB variant of Tysachus disease, respectively).

[0026] Figure 9A and 9B The effects of acetyl-DL-leucine treatment over time on the overall clinical severity score (CSS) and overall annual severity increment score (ASIS) of 10 NPC patients were shown.

[0027] Figure 10A-10J The effect of acetyl-DL-leucine treatment on CSS sub-scores over time was shown for each of the ten NPC patients. Detailed Implementation

[0028] Racemic forms of acetyl-leucine (acetyl-DL-leucine) and their salts are effective in treating vertigo of various origins, especially Meniere's vertigo and vertigo of inflammatory (vestibular neuritis) or toxic origins. For example, acetyl-leucine is marketed in racemic form by Pierre Fabre Medicament as an antivertigo drug under the brand name Tanganil®. Clinical results reported by various authors for Tanganil® show improvement in vertigo symptoms, including the disappearance of vertigo attacks, in over 95% of cases.

[0029] Acetyl-DL-leucine has been used in France since 1957 to treat acute vertigo with a good safety profile, but its long-term safety remains undetermined. Despite numerous hypotheses, including membrane potential stability, its pharmacological and electrophysiological mechanisms of action remain unclear. (Vibert et al. (2001)) Eur J Neurosci ; 13(4): 735-48; Ferber-Viart et al. (2009) Audiol Neurootol ; 14(1): 17-25). FDG-µPET study in a rat model of acute unilateral labyrinthectomy (Zwergal et al. (2016)). Brain Struct Funct ; 221(1):159-70) showed that the L enantiomer, N-acetyl-L-leucine, has a significant effect on postural compensation caused by vestibulocerebellar activation and posterolateral thalamic inactivation (Gunther et al. (2015)). PLoS One ; 10(3): e0120891). The improvement of symptoms of cerebellar ataxia by the use of acetyl-DL-leucine has been shown in a series of cases of cerebellar patients (Strupp et al. (2013)). J Neurol ; 260(10): 2556-61). Another series of cases found no benefit (Pelz et al. (2015)). J Neurol ;262(5): 1373-5). Quantitative gait analysis showed that acetyl-DL-leucine improved temporal gait variability in patients with cerebellar ataxia (Schniepp et al. (2015)). Cerebellum ; 3:8). A one-month study involving 12 patients with Niemann-Pick disease type C (NPC) showed improvement in ataxia symptoms (Bremova et al. (2015)). Neurology; 85(16): 1368-75). Furthermore, PET studies in ataxia patients given acetyl-DL-leucine showed increased metabolism in the midbrain and lower brainstem of the responders (Becker-Bense et al. (2015)). Abstract EAN ).

[0030] However, the use of acetyl-leucine for the treatment of LSD, which typically progresses over years to decades, is unknown. This disclosure surprisingly demonstrates that acetyl-leucine, or a pharmaceutically acceptable salt thereof, can be used to treat LSD in subjects in need, for example, 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 delaying or reversing (e.g., for a prolonged period) the progression of LSD or one or more symptoms of LSD compared to typical disease progression. These exemplary applications of this disclosure, as well as other applications described herein, are entirely unexpected because these benefits have not been previously observed and cannot be inferred, nor are they teachings of the prior art. LSD is one of the heterogeneous groups of hereditary diseases characterized typically by the accumulation of undigested or partially digested macromolecules, leading to cellular dysfunction (e.g., increased lysosomal volume compared to healthy subjects) and clinical abnormalities. As demonstrated by the examples, but not wishing to be bound by any specific theory, the inventors have particularly found that acetyl-leucine can improve cellular dysfunction (e.g., by reducing lysosomal volume to control values) and clinical abnormalities in subjects with LSD.

[0031] Therefore, this 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.

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

[0033] As used herein, "subject" can be a vertebrate, mammal, or livestock. Therefore, the compositions according to this disclosure can be used to treat any mammal, such as livestock (e.g., horses, cattle, sheep, or pigs), pets (e.g., cats, dogs, rabbits, or guinea pigs), laboratory animals (e.g., mice or rats), or for other veterinary applications. For example, the subject is a human.

[0034] As used in this article, the singular forms “a,” “one,” and “the” include the plural forms.

[0035] The terms “approximately” and “about” refer to a quantity that is nearly identical to a reference number or value, including a quantity measured with an acceptable degree of error given its properties or measurement precision. As used herein, the terms “approximately” and “about” should generally be understood to include ±20% of the specified quantity, frequency, or value. Unless otherwise stated, the numerical values ​​given herein are approximate and are meant to be inferred from the terms “approximately” or “about” in the absence of explicit specification.

[0036] As used herein, the term “application” means (1) the combination provided, given, administered, and / or prescribed by a healthcare professional or their authorized agent or under their guidance, in accordance with the present disclosure, and (2) the combination administered, taken, or consumed by a patient or himself / herself according to the present invention.

[0037] The text refers to "acetyl-leucine" throughout, including its pharmaceutically acceptable salts, even if not explicitly stated otherwise.

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

[0039] The term "pharmaceutically acceptable salt" as used in this article refers to any salt preparation suitable for pharmaceutical use. Pharmaceutically acceptable salts include, but are not limited to, amine salts, such as N,N'-dibenzylethylenediamine, chloroprocaine, choline, ammonia, diethanolamine and other hydroxyalkylamines, ethylenediamine, N-methylglucosamine, procaine, N-benzylphenethylamine, 1-p-chlorobenzyl-2-pyrrolidone-1'-ylmethylbenzimidazole, diethylamine and other alkylamines, piperazine, tris(hydroxymethyl)aminomethane, etc.; alkali metal salts, such as lithium, potassium, sodium, etc.; alkaline earth metal salts, such as barium, calcium, magnesium, etc.; transition metal salts, such as zinc, aluminum, etc.; other metal salts, such as sodium hydrogen phosphate, disodium phosphate, etc.; inorganic acids, such as hydrochlorides, sulfates, etc.; organic acid salts, such as acetates, lactates, malates, tartrates, citrates, ascorbic acid salts, succinates, butyrates, valerates, and fumarates, etc.

[0040] Acetyl-leucine or a pharmaceutically acceptable salt thereof may be formulated and administered to a subject in accordance with teachings known in the art. For example, acetyl-leucine, or a pharmaceutically acceptable salt thereof, may be formulated into a pharmaceutical composition. A pharmaceutical composition may comprise acetyl-leucine, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. References to pharmaceutical compositions may include an active agent alone or an active agent in the form of a pharmaceutical composition.

[0041] The pharmaceutical composition can take any of a variety of different forms, depending particularly on its manner of administration. Thus, for example, it can be in the form of powder, tablet, capsule, liquid, ointment, cream, gel, hydrogel, aerosol, spray, micelle solution, transdermal patch, liposome suspension, or any other suitable form that can be administered to a person or animal in need of treatment.

[0042] The term "pharmaceutically acceptable carrier" as used herein refers to any known compound or combination of known compounds known to those skilled in the art for use in the formulation of pharmaceutical compositions. It should be understood that the carrier of a pharmaceutical composition should be a substance that can be tolerated by the subject to whom it is administered.

[0043] In one embodiment, the pharmaceutically acceptable carrier may be a solid, and the composition may be in the form of a powder or tablet. Solid pharmaceutically acceptable carriers may include, but are not limited to, one or more substances that can be flavoring agents, buffers, lubricants, stabilizers, solubilizers, suspending agents, wetting agents, emulsifiers, dyes, fillers, flow aids, compression aids, inert binders, sweeteners, preservatives, coatings, or tablet disintegrants. The carrier may also be an encapsulating material. In powders, the carrier may be a finely ground solid premixed with a finely ground active agent according to the invention. In tablets, the active agent may be mixed with a carrier having the necessary tableting properties in an appropriate proportion and compressed into the desired shape and size. For example, powders and tablets may contain up to 99% active agent. Suitable solid carriers include, for example, calcium phosphate, magnesium stearate, talc, sugar, lactose, dextrin, starch, gelatin, cellulose, polyvinylpyrrolidone, low-melting-point waxes, and ion exchange resins. In another embodiment, the pharmaceutically acceptable carrier may be a gel, and the composition may be in the form of a cream, etc.

[0044] The carrier may include, but is not limited to, one or more excipients or diluents. Examples of such excipients include gelatin, gum arabic, lactose, microcrystalline cellulose, starch, sodium glycolate starch, dicalcium phosphate, magnesium stearate, talc, and colloidal silica.

[0045] In another embodiment, the pharmaceutically acceptable carrier can be a liquid. In one embodiment, the pharmaceutical composition is in solution form. Liquid carriers are used to prepare 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 a pharmaceutically acceptable oil or fat. Liquid carriers may contain other suitable pharmaceutical additives, such as solubilizers, emulsifiers, buffers, preservatives, sweeteners, flavorings, suspending agents, thickeners, colorants, viscosity modifiers, stabilizers, or osmotic pressure modifiers. Suitable examples of liquid carriers for oral and parenteral administration include water (partially containing the above-mentioned additives, such as cellulose derivatives, like sodium carboxymethyl cellulose solution), alcohols (including monohydric and polyhydric alcohols, such as diols) and their derivatives, and oils (e.g., fractionated coconut oil and peanut oil). For parenteral administration, the carrier can also be an oily ester, such as ethyl oleate and isopropyl myristate. Sterile liquid carriers can be used in sterile liquid form for parenteral administration of compositions. The liquid carrier used in the pressurized composition may be a halocarbon or other pharmaceutically acceptable propellant.

[0046] Liquid pharmaceutical compositions, as sterile solutions or suspensions, can be administered, for example, via 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 suitable sterile injection media.

[0047] The composition can be administered orally in the form of a sterile solution or suspension containing other solutes or suspending agents (e.g., sufficient saline or glucose to make the solution isotonic), bile salts, gum arabic, gelatin, sorbitan monooleate, polysorbate 80 (oleate of sorbitol and its anhydride copolymerized with ethylene oxide), etc. The composition can also be administered orally in liquid or solid form. 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 for parenteral administration include sterile solutions, emulsions, and suspensions.

[0048] Acetyl-leucine and compositions containing it can alternatively be administered by inhalation (e.g., intranasal). The 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-release or delayed-release devices. Such devices can be inserted, for example, onto or under the skin, and the drug can be released over weeks or even months. Such devices may be advantageous when long-term treatment with acetyl-leucine according to this disclosure is required (typically requiring frequent administration, e.g., at least daily)).

[0050] In one embodiment, the pharmaceutical composition is in tablet form. In the tablet, the active agent may be mixed in an appropriate proportion with a carrier having the necessary compressibility properties, such as a pharmaceutically acceptable carrier, and compressed into the desired shape and size. The tablet may 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 solid dosage forms suitable for oral administration, particularly in tablet form.

[0052] Pharmaceutical compositions in solid oral dosage forms, such as tablets, can be prepared by any method known in the pharmaceutical field. Pharmaceutical compositions are typically prepared by mixing acetyl-leucine or a pharmaceutically acceptable salt thereof with a conventionally pharmaceutically acceptable carrier.

[0053] Tablets can be formulated as known in the art. For example, Tanganil. ® Wheat starch, pregelatinized corn starch, calcium carbonate, and magnesium stearate are included as excipients. The same or similar excipients may be used in conjunction with this disclosure, for example.

[0054] Each 700mg Tananil® tablet contains the following: 500mg acetyl-DL-leucine, 88mg wheat starch, 88mg pregelatinized corn starch, 13mg calcium carbonate, and 11mg magnesium stearate. The same tablets may be used in conjunction with this disclosure, for example.

[0055] This disclosure describes acetyl-leucine, including compositions and methods thereof, for treating LSD or one or more symptoms of LSD in a subject of need. The subject of need may have genetic, biochemical, or other similar identifiable markers of LSD. For example, a marker of LSD might be a cellular marker. The subject of need may have been diagnosed with LSD. For example, the subject may have been diagnosed with LSD based on genetic, biochemical, or other similar identifiable markers. The subject of need may be suspected of having or at risk of having LSD. For example, the subject may have a genetic predisposition to LSD (e.g., the subject may have one or more family members with LSD). The subject of need may have symptoms (i.e., have one or more LSD-related symptoms). The subject of need may be asymptomatic. The terms “symptomatic” and “asymptomatic” should be understood to be used when referring to symptoms of LSD. Subjects with genetic, biochemical, or other similar identifiable markers of LSD, such as those diagnosed with LSD based on genetic, biochemical, or other similar identifiable markers but without further symptoms, are included in the scope of “asymptomatic” for the purposes of this disclosure.

[0056] As used herein, "treating LSD or one or more symptoms of LSD" refers to delaying the onset of LSD or one or more symptoms of LSD (the disease or symptoms that would normally be expected to develop according to typical disease progression), reducing the severity of LSD, or reducing or eliminating one or more existing symptoms associated with LSD, delaying the progression of LSD or one or more symptoms of LSD over time compared to typical disease progression, and / or reversing the progression of progressive LSD or one or more symptoms of LSD over time. "Treating LSD or one or more symptoms of LSD" may also be referred to as improving biochemical markers of LSD.

[0057] As used herein, "typical disease progression," "typically expected disease progression," etc., refer to the typical or expected progression of LSD, one or more symptoms associated with LSD, or biochemical markers of LSD if the subject is untreated. Typical or expected disease progression can be based, for example, on known scales, indices, ratings, or scores, or other suitable tests used to assess the progression of LSD, one or more symptoms associated with LSD, or biochemical markers of LSD, such as those described as examples herein. Scales, indices, ratings, or scores, or other suitable tests 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 LSD or LSD-related symptoms or sets of symptoms. Typical or expected disease progression can be determined on a subject-by-subject basis, or it can be determined based on what is typically observed or experienced in a set of subjects affected by LSD, such as a group or subgroup of subjects. Subgroups may include, for example, subgroups of the same sex, subgroups of the same or similar age, subgroups of the same or similar onset times of one or more symptoms, etc.

[0058] In one implementation, “treating LSD or one or more symptoms of LSD” means delaying the onset of LSD or one or more symptoms of LSD (the disease or symptoms that would normally be expected to occur according to typical disease progression). As used herein, “delaying the onset of LSD or one or more symptoms of LSD” and the like means increasing the time before the onset of LSD or one or more symptoms of LSD, or preventing its onset. For example, an onset can be said to be delayed when the time before the manifestation of LSD or one or more symptoms of LSD is at least 5% longer than the time observed according to typical disease progression. Further, for example, the observed time increase is 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 implementation, the subject is asymptomatic. Administration of acetyl-leucine can be initiated during the asymptomatic period in the subject to delay the onset of LSD or one or more symptoms of LSD (the disease or symptoms that would normally be expected to occur according to typical disease progression). In another implementation, the subject has symptoms. Acetyl-leucine administration can be initiated when the subject has some symptoms to delay the onset of one or more other symptoms of LSD (the disease or symptoms that would normally be expected to occur according to typical disease progression). Subjects who require treatment can continue acetyl-leucine therapy as described herein. In one embodiment, the treatment prevents the onset of one or more symptoms of LSD (the disease or symptoms that would normally be expected to occur according to typical disease progression).

[0059] In one implementation, “treating LSD or one or more symptoms of LSD” means reducing the severity of LSD, or reducing or eliminating one or more existing symptoms associated with LSD. The severity of LSD or existing symptoms can be assessed using known scales, indices, ratings, or scores, such as those described herein as examples, or using another suitable test for assessing severity. For example, scales, indices, ratings, or scores, or other suitable tests, may correspond to the overall severity of the disease or the severity of one or more symptoms associated with the disease. In one implementation, the treatment improves this assessment from a value or severity characteristic of symptomatic patients to a value or severity characteristic of non-symptomatic patients.

[0060] In one implementation, “treating LSD or one or more symptoms of LSD” means delaying or reversing the progression of LSD or one or more symptoms associated with LSD compared to typical disease progression. The duration of delay or reversal may be consistent with the treatment duration described herein. Treatment may delay or reverse progression for a duration of, for example, more than 7 days, more than 2 weeks, more than 3 weeks, more than 1 month, more than 6 weeks, more than 7 weeks, or more than 2 months. For example, treatment may delay or reverse progression for a duration of more than 3 months, more than 4 months, more than 5 months, or more than 6 months. It may delay or reverse progression for a duration of, for example, more than 1 year, more than 2 years, more than 3 years, more than 4 years, more than 5 years, or more than 10 years. Treatment may delay or reverse the progression of LSD or one or more symptoms associated with LSD throughout the patient's life.

[0061] In one implementation, “treating LSD or one or more symptoms of LSD” means delaying the progression of LSD or one or more symptoms of LSD over time compared to typical disease progression. As used herein, “delaying the progression of LSD or one or more symptoms of LSD over time” and similar expressions mean 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 increase in severity of the disease or one or more symptoms of the disease). Disease progression can be determined, for example, using known scales, indices, ratings, or scores, such as those described herein as examples, or using another suitable test for assessing progression. For example, a scale, index, rating, or score, or other suitable test, may correspond to overall disease progression or to the progression of one or more symptoms associated with the disease. In one implementation, “delaying the progression of LSD or one or more symptoms of LSD” means that the severity value of the subject’s disease (e.g., overall severity or severity of one or more symptoms) determined by a scale, index, rating, or score, or other suitable test for assessing severity, does not increase meaningfully (e.g., remains at least substantially unchanged). In one implementation, “delaying the progression of LSD or one or more symptoms of LSD” means preventing a subject from reaching a severity value compared to a value corresponding to typical disease progression, or increasing the time it takes for a subject to reach a severity value (e.g., reducing the rate of change in severity increase), said severity value being determined based on a known scale, index, rating, or score, or another 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 the time observed according to typical disease progression. Further, for example, the observed time increase is 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 treatment delaying the progression of LSD or one or more symptoms of LSD may be consistent with the treatment duration described herein. In one implementation, 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 implementation, 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. Treatment can delay progression throughout a patient's life.

[0062] In one implementation, “treating LSD or one or more symptoms of LSD” means 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” means stopping the progression of the disease or one or more symptoms of the disease and reducing the severity of the disease or one or more symptoms of the disease over time. Disease progression and severity granularity are determined, for example, using known scales, indices, ratings, or scores, such as those described herein as examples, or using another suitable test for assessing progression and severity. For example, scales, indices, ratings, scores, or other suitable tests may correspond to the overall progression and severity of the disease, or to the progression and severity of one or more symptoms associated with the disease. In one implementation, “reversing the progression of LSD or one or more symptoms of LSD over time” indicates that the disease severity value (e.g., overall severity or severity of one or more symptoms) determined by the subject using known scales, indices, ratings, or scores, or other suitable tests for assessing severity, improves over time (i.e., shows a decrease in severity over time). The duration of treatment to reverse the progression of LSD or one or more symptoms of LSD may be consistent with the treatment duration described herein. In one implementation, the treatment reverses 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 implementation, the treatment reverses 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 reverse progression throughout the patient's life.

[0063] In one embodiment, “treating LSD or one or more symptoms of LSD” refers to improving a subject’s biochemical markers of LSD (e.g., increasing levels of stored metabolites or secondary biochemical changes resulting from primary storage). Biochemical markers are signals of disease activity and can provide a sustained indication of disease severity and progression over time. In one embodiment, the biochemical marker improves relative to a control value. In one embodiment, the biochemical marker is selected from increased lysosomal volume and increased glycosphingolipid (GSL) levels. In one embodiment, the biochemical marker is increased lysosomal volume, and the treatment reduces the subject’s lysosomal volume. In one embodiment, the biochemical marker is increased glycosphingolipid (GSL) levels, and the treatment reduces the subject’s GSL levels. In one embodiment, the treatment improves the biochemical marker over time. For example, in one embodiment, improving the biochemical marker over time means that, compared to typical disease progression, the treatment improves the biochemical marker toward a control value over time, prevents the biochemical marker from progressing over time, and / or delays the biochemical marker’s progression over time. The time it takes for the treatment to improve the biochemical marker may be consistent with the treatment duration described herein. In one embodiment, the treatment improves the biochemical markers 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 markers 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 markers throughout the patient's life.

[0064] The term "symptoms" for LSD includes any clinical or laboratory manifestations 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 and psychiatric symptoms. Examples of neurological symptoms include ataxia, other motor disorders such as hypokinesis, rigidity, tremor, or dystonia, central oculomotor disorders such as vertical and horizontal supranuclear saccades / gaze palsy, and neuropsychiatric deficits such as dementia. Examples of psychiatric symptoms include depression, behavioral disorders, or psychosis. Symptoms may occur 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,” Am J Med Genet Part B 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] Therefore, “treating LSD or one or more symptoms of LSD” can be equivalent to achieving an assessment of improvement, such as the SARA, SCAFI, ICARS, BARS, mDRS, EQ-5D-5L, VAS, WAIS-R, WISC-IV, CSS, and / or MoCA scores described herein, or an assessment of improvement in the results of another test suitable for characterizing LSD subjects. For example, in one implementation, “reducing the severity of LSD, or reducing the severity of one or more existing symptoms of LSD, or eliminating one or more existing symptoms of LSD” means improving the results of SARA, SCAFI, ICARS, BARS, mDRS, EQ-5D-5L, VAS, WAIS-R, WISC-IV, CSS, and / or MoCA scores, or the results of another test suitable for assessing severity, such as a score or result that transforms the severity value characteristic of symptomatic subjects to the value characteristic of non-symptomatic subjects. In another implementation, “delaying the progression of LSD or one or more symptoms of LSD” means that the subject’s SARA, SCAFI, ICARS, BARS, mDRS, EQ-5D-5L, VAS, WAIS-R, WISC-IV, CSS, and / or MoCA scores, or the results of another suitable test for assessing progression, do not increase significantly (e.g., remain at least substantially unchanged). In yet another implementation, “delaying the progression of LSD or one or more symptoms of LSD” means preventing the subject’s SARA, SCAFI, ICARS, BARS, mDRS, EQ-5D-5L, VAS, WAIS-R, WISC-IV, CSS, and / or MoCA scores, or the results of another suitable test for assessing progression, from reaching values ​​comparable to typical disease progression, or increasing the time to reach those values. In another implementation, “reversing the progression of LSD or one or more symptoms of LSD over time” means that the subject’s SARA, SCAFI, ICARS, BARS, mDRS, EQ-5D-5L, VAS, WAIS-R, WISC-IV, CSS and / or MoCA scores, or the results of another appropriate test used to assess progression, improve over time (i.e., show a decrease in severity over time).

[0067] For example, to assess overall neurological status, the mDRS and four-domain scales (gait, manipulation, language, and swallowing) can be used. Cerebellar function can be assessed using the SARA and SCAFI, the 9-hole plate test (9HPT), and the number of PATA repetitions within 10 seconds. The SARA is an eight-item clinical rating scale (gait, standing, sitting, language, fine motor function, and kinesiology; range 0-40, where 0 is the best neurological state and 40 is the worst). The SCAFI includes an 8-meter walking time (8MW; performed by having the patient walk from one line to another as quickly as possible twice (excluding turns). Subjective impairment and quality of life can be assessed using the EQ-5D-5L assessment questionnaire and VAS evaluation. To assess oculomotor function, [the following can be done]. The peak velocity of saccades, smooth tracking gain, peak slow-phase velocity of fixation-induced nystagmus (gaze hold function), peak slow-phase velocity of optomotor nystagmus, and level vestibular reflex gain are measured using 3D video eye-tracking (EyeSeeCam). For cognitive assessment, WAIS-R or WISC-IV and MoCA can be used to assess different cognitive domains, including attention and concentration, executive function, memory, language, visual-structural skills, conceptual thinking, calculation, and orientation, with a maximum of 30 points and a cutoff score of 26. Technicians will know how to perform these tests and others like them.

[0068] Acetyl-leucine or a pharmaceutically acceptable salt thereof may be administered, for example, at doses of about 500 mg to about 15 g / day or about 500 mg to about 10 g / day, for example, about 1.5 g to about 10 g / day, optionally via solid or liquid oral routes. Acetyl-leucine or a pharmaceutically acceptable salt thereof may be administered, for example, according to Tanganil... ® The dosage for adults is 1.5 g to 2 g per day, 3-4 tablets, divided into two doses in the morning and evening.

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

[0070] In one embodiment, the dosage is about 1 g to about 15 g / day, about 1 g to about 10 g / day, or about 1.5 g to about 7 g / day. It can be about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 g to about 15 g / day. It can be about 2, 3, 4, 5, 6, 7, 8, or 9 g to about 10 g / day. It can be more than about 1.5 g / day, but less than about 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, or 5 g / day. In one embodiment, the dosage is about 4 g to about 6 g / day. In one embodiment, the dosage is about 4 g to about 5 g / day. In one embodiment, the dosage is about 4.5 g / day. In one embodiment, the dosage is about 5 g / day. In one embodiment, these dosages are administered in a solid oral dosage form, particularly tablets. In another embodiment, when in racemic form, these dosages are for acetyl-leucine. When an enantiomer is in excess, the dose of acetyl-leucine may be lower than those described herein, for example, by about 50%. Therefore, this disclosure also explicitly covers the above-mentioned dose range when the dose is halved.

[0071] The total daily dose can be distributed across multiple administrations, meaning that administration can be performed two or more times per day to reach the total daily dose. For example, the number of tablets required to provide the total daily dose of acetyl-leucine can be divided into two administrations (e.g., morning and evening) or three administrations (e.g., morning, noon, and evening). Each administration can be taken with or without food as appropriate. 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 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 given as 3 Tanganil tablets before, during, or after breakfast. ® (or an equal amount), take 3 more tablets before, during or after lunch, and another 3 tablets before, during or after dinner.

[0072] Acetyl-leucine administration according to this disclosure can be initiated before or after the discovery of genetic, biochemical, or other similar identifiable markers of LSD in a subject, for example, in the former case, when the subject is suspected of having or at risk of having LSD. Administration can be initiated at or near the time the subject is found to have genetic, biochemical, or other similar identifiable markers of LSD. Similarly, administration can be initiated before, at, near, or after the subject is diagnosed with LSD, or before, at, near, or after the discovery of genetic, biochemical, or other similar identifiable markers of LSD in a subject. Acetyl-leucine can be initiated whether the subject has symptoms or not. In particular, one of the advantages of treatment with acetyl-leucine according to this disclosure is that administration of acetyl-leucine can be initiated as early as when the subject is found to have genetic and / or biochemical markers of LSD, but before the subject displays symptoms of LSD (symptoms other than genetic and / or biochemical markers, i.e., the subject is asymptomatic) or before the subject displays markers considered to be indicative of the disease. Treatment may delay the onset of LSD or one or more symptoms associated with LSD, as described herein. Treatment may also last for the duration described in this article.

[0073] As discussed herein, an advantage of treatment with acetyl-leucine according to this disclosure is that acetyl-leucine can be administered for extended periods to delay or even reverse the progression of LSD or one or more symptoms of LSD in a subject, compared to typical disease progression. Treatment duration can be, for example, 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. Treatment duration can also 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. Treatment duration may be the patient's lifespan.

[0074] Any and all combinations of dosage form, dosage, administration regimen, and duration of treatment are contemplated and covered by this invention. In one embodiment, the dosage is about 4 g to about 10 g / day, divided into 1, 2, or 3 doses / day, for a duration of about 2 months or more. In another embodiment, the dosage is more than 4 g but not more than 5 g / day, divided into 1, 2, or 3 doses / day, for a duration of about 6 months or more. The dosage form may be a solid oral dosage form, particularly tablets.

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

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

[0077] The sphingolipid storage disease can be selected from Niemann-Pick disease type A / B, Gaucher disease type I, II, and III, Krab disease, Fabry disease, Schindler disease, GM1 ganglioside storage disease, Morquio B disease, GM2 ganglioside storage disease, metachromatic leukodystrophy, Fabry disease, multiple sulfatase deficiencies, lysosomal acid lipase deficiency, and galacturonic acidosis. In one embodiment, the sphingolipid storage disease is selected from Niemann-Pick disease type A, GM1 ganglioside storage disease, Tessachs disease, AB variants of Tessachs disease, and Sandhof disease.

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

[0079] The mucopolysaccharide storage disease can be selected from any one of MPS IH, MPS IHS, 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] Oligosaccharide storage disorder can be selected from β-mannoside storage disorder, α-fucoside storage disorder, and aspartic glucosamineuria. In one embodiment, the oligosaccharide storage disorder is aspartic glucosamineuria.

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

[0082] Glycogen storage disorders can be selected from Pompe disease that occurs in infancy, late-onset Pompe disease, and Danon disease.

[0083] Lysosomal transport disorders can be selected from cystinosis, osteogenesis imperfecta, sialic acid storage disorders, and infantile free sialic acid storage disorders.

[0084] LSD can be a deficiency of primary lysosomal hydrolases, a deficiency of post-translational processing of lysosomal enzymes, a deficiency of transport of lysosomal enzymes, a deficiency of protection of lysosomal enzymes, a deficiency of soluble non-enzymatic lysosomal proteins, a deficiency of transmembrane (non-enzymatic) proteins, or an unclassified deficiency.

[0085] In one implementation, LSD is selected from primary lysosomal hydrolases that are deficient. Primary lysosomal hydrolases deficiencies include, but are not limited to, Taysaks disease (β-aminohexosidase A deficiency), Sandhof disease (β-aminohexosidase A+B deficiency), Fabry disease (α-galactosidase A deficiency), Clapham disease (β-galactosylceramide deficiency), Niemann-Pick disease type A and B (sphingomyelinase deficiency), metachromatic leukodystrophy (aromatic sulfatase A deficiency), MPS IH (Heller syndrome; α-iduronidase deficiency), MPS IS (Schner-Ellison syndrome; α-iduronidase deficiency), MPS IH-S (Heller-Schner syndrome; α-iduronidase deficiency), MPS II (Hunter syndrome; iduronidase deficiency), MPS IIIA (Samfelipo syndrome; sulfaacetylheparinase deficiency), MPS IIIB (Samfelipo syndrome; acetylα-glucosidase deficiency), and MPS IIIC. (Samfelipo C syndrome; acetyl-CoA: α-glucosinolate N-acetyltransferase deficiency), MPS IIID (Samfelipo D syndrome; N-acetylglucosamine-6-sulfatase deficiency), MPSIV A (Morquio A disease; acetylgalactosamine-6-sulfatase deficiency), MPS IVB (Morquio B disease; β-galactosidase deficiency), MPS V (renamed MPS IS), MPS VI (Maroteaux Lamy Syndrome; acetylgalactosamine-4-sulfatase (aromatic sulfatase B) deficiency), MPS VII (Sly syndrome; β-glucuronidase deficiency), MPS IX (hyaluronidase deficiency), Wollman / cholesterol ester storage disease (WD; acid lipase deficiency), Pompe disease (type II; α-glucosinolate ester storage disease). 1,4-glucosidase deficiency), aspartic glucuria (glycosylasparaginase deficiency), fucoside storage disease (α-fucoside deficiency), α-mannoside storage disease (α-mannoside deficiency), β-mannoside storage disease (β-mannoside deficiency), Sheldr's disease (N-acetylgalactosidase deficiency), sialic acid storage disease / ML Type I (α-neuraminidase deficiency), Infant neuron-like lipofuscin storage disease (CLN1; palmitoyl protein thioesterase deficiency), Late-stage infantile neuron-like lipofuscin storage disease (CLN2; carboxypeptidase deficiency), Early-stage infantile GM1 ganglioside storage disease, Late-stage infantile GM1 ganglioside storage disease, Adult-onset GM1 ganglioside storage disease, Type 1 Gaucher disease (non-neuropathy), Type 2 / 3 Gaucher disease (neuropathy), Type 4 neuronal eosinophilic lipofuscin storage disease (CLN4; Kuff's disease; Adult NCL; palmitoyl protein thioesterase-1 deficiency (type A));The deficiencies include cathepsin F deficiency (type B), neuronal eosinophilic lipofuscinosis type 10 (CLN10; congenital cathepsin D deficiency), osteogenesis imperfecta (cathepsin K deficiency), infantile-onset Pompe disease, late-onset Pompe disease, Fabry disease (Fabry lipogranulomatosis; ceramide deficiency; fibroblastic mucopolysaccharidosis; lipogranulomatosis), and galacturonic acidosis (protective protein cathepsin A deficiency, PPCA deficiency). In one embodiment, the primary lysosomal hydrolase deficiency is selected from Taysaks disease, Sandhof disease, Niemann-Pick disease type A, Niemann-Pick disease type B, neuronal ceroid lipofuscinosis, Gaucher disease, Fabry disease, Krabby disease, GM1 ganglioside storage disease, GM2 ganglioside storage disease, metachromatic leukodystrophy, and Fabry disease. In one embodiment, the primary lysosomal hydrolase deficiency is selected from Taysaks disease, Sandhoff's disease, Niemann-Pick disease type A, Niemann-Pick disease type B, and GM1 ganglioside storage disease.

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

[0087] In one embodiment, LSD is selected from lysosomal enzyme transport defects. Lysosomal enzyme transport defects include, but are not limited to, type II mucolipidemia (I-cell disease; N-acetylglucosamine phosphotransferase deficiency), IDA type mucolipidemia (pseudo-Hurler's multiple malnutrition; N-acetylglucosamine phosphotransferase deficiency), and type IIIC mucolipidemia.

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

[0089] In one implementation, LSD is a soluble non-enzymatic lysosomal protein defect. Soluble non-enzymatic lysosomal protein defects include, but are not limited to, GM2 activator protein defect (AB variant), Niemann-Pick disease type C2 (NPC2), and sphingolipid activator protein (SAP) defect.

[0090] In one embodiment, LSD is a transmembrane (non-enzymatic) protein deficiency. Transmembrane (non-enzymatic) protein deficiencies include, but are not limited to, Danon disease (lysosome-associated membrane protein 2 (LAMP2) deficiency), NPC (NPC1 deficiency), cystinosis (cystine deficiency), infantile free sialic acid storage disease (ISSD; sialic acid transporter deficiency), Salla disease (free sialic acid storage disease; sialic acid transporter deficiency), adolescent neuron-like lipofuscin storage disease (CLN3, Battendisease)), adult neuron-like lipofuscin storage disease (Kufs disease; adult NCL; palmitoyl thioesterase-1 deficiency (type A); cathepsin F deficiency (type B)), neuronal ceroid lipofuscin storage disease (NCL) (CLN6, CLN7, and LN8), and type IV mucolipin storage disease (mucoprotein deficiency). In one embodiment, LSD is Niemann-Pick disease type C1 or Niemann-Pick disease type C2.

[0091] In one implementation, LSD is an unclassified defect. Unclassified defects include, but are not limited to, neuronal cerebrolipofuscin storage disease (NCL) (CLN5 and CLN9).

[0092] LSD to be treated by the compositions and methods of the present invention can be any of neuronal cerebrolipofuscin storage disease, primary glycosphingolipid diseases (i.e., Gaucher disease, Fabry disease, GM1, GM2 ganglioside storage disease, Krabby disease, and metachromatic leukodystrophy (MLD)), Fabry disease, and multiple sulfatase deficiencies. In one embodiment, LSD involves significant central nervous system (CNS) involvement. For example, LSD can be selected from NPC, Tessachus disease, Sandhoff disease, GM1 ganglioside storage disease, or Fabry disease.

[0093] In one embodiment, LSD is Niemann-Pick disease type A. In another embodiment, LSD is Niemann-Pick disease type B. In yet another embodiment, LSD is Niemann-Pick disease type C (C1 or C2). Niemann-Pick disease is a heterogeneous group of autosomal recessive LSD. Common cellular features include abnormal sphingomyelin (SM) storage in mononuclear phagocytes and parenchymal tissues, as well as (liver)splenomegaly. Among the three major subgroups (AC), NPC (formerly classified as NPC and NPD, now considered a single disease) is classified as lethal neuroendocrine LSD caused by the accumulation of unesterified cholesterol induced by abnormal intracellular cholesterol transport in the late endosome / lysosomal compartment. Outside the CNS, cellular features of NPC include abnormal accumulation of unesterified cholesterol and other lipids (such as GSL) in the late endosome / lysosomal compartment. Conversely, there is no net increase in cholesterol in the CNS (although it does have an altered distribution) but highly elevated GSL levels are present. The progressive neurodegenerative disease is characterized by the continuous degeneration of GABAergic cerebellar Purkinje neurons, which parallels the occurrence and development of other aspects of neurological dysfunction that occur during NPC. Genetic studies have shown that NPC 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 a mystery. NPC1 encodes a multimembrane-crossing protein that restricts the late endosome / lysosome, while NPC2 is a lysosomal cholesterol-binding protein. When NPC1 is inactivated, sphingosine is the first lipid to be stored, suggesting that NPC1 plays a role in the transport of sphingosine in lysosomes, where sphingosine is normally produced as part of sphingolipid catabolism. Elevated sphingosine, in turn, leads to a defect in calcium entry into the acidic reservoir, resulting in a significant reduction in calcium release from this chamber. This, in turn, prevents late endosome-lysosome fusion, a calcium-dependent process, and leads to a secondary accumulation of lipids (cholesterol, sphingomyelin, and glycosphingolipids), cargoes transported via the late endocytic pathway. Other secondary consequences of inhibiting NPC1 function include defective endocytosis and failure to clear autophagic vacuoles. The NPC1 / NPC2 cellular pathway has been shown to be targeted by pathogenic mycobacteria to promote their survival in late endosomes.

[0094] Tessachs disease is a fatal, inherited lipid metabolism disorder characterized specifically in the CNS tissues, caused by a deficiency of the A isoenzyme of β-aminohexosinosinase. Mutations in the HEXA gene, encoding the α subunit of β-hexosinosinase, lead to the deficiency of the A isoenzyme. Tessachs is the prototype of a group of disorders characterized by GM2 ganglioside storage disorders, which are characterized by defects in GM2 ganglioside degradation. GM2 gangliosides (monosialidoganglioside 2) begin accumulating in neurons during fetal life.

[0095] Sandhoff's disease is caused by a deficiency of the A and B (basic) isoenzymes of β-hexosaminease. Mutations in the HEXB gene, which encodes the β subunit of β-hexosaminease, result in a deficiency of the B isoenzyme.

[0096] GM1 ganglioside storage disorder is caused by a deficiency of β-galactosidase, which leads to lysosomal storage of GM1 ganglioside (monosialized ganglioside 1).

[0097] Fabry disease is caused by a deficiency of α-galactosidase, which leads to the lysosomal accumulation of ceramide trihexoside.

[0098] In one embodiment, LSD is selected from Tessachus disease, AB variant of Tessachus disease, Sandhoff disease, Niemann-Pick disease type A, type II mucoidosis, type III mucoidosis, MPS III, MPS VII, GM1 gangliosidosis, and aspartic glucosamineuria. In one embodiment, LSD is Sandhoff disease. In one embodiment, LSD is Tessachus disease. In one embodiment, LSD is AB variant of Tessachus disease. In one embodiment, LSD is type II mucoidosis. In one embodiment, LSD is type III mucoidosis. In one embodiment, LSD is GM1 gangliosidosis. In one embodiment, LSD is MPS III. In one embodiment, LSD is MPS VII. In one embodiment, LSD is type A Niemann-Pick disease. In one embodiment, LSD is aspartic glucosamineuria.

[0099] In one implementation, LSD is not Niemann-Pick disease. In one implementation, LSD is not Niemann-Pick type C disease.

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

[0101] In one implementation, acetyl-leucine or a pharmaceutically acceptable salt thereof treats gait deterioration, motor function deterioration, and / or motor decline associated with Sandhoff's disease. For example, acetyl-leucine or a pharmaceutically acceptable salt thereof may delay the onset of gait deterioration, motor function deterioration, and / or motor decline associated with Sandhoff's disease, reduce its severity or eliminate it, or delay or reverse its progression.

[0102] In one implementation, acetyl-leucine or a pharmaceutically acceptable salt thereof treats decreased coordination, tremor, motor impairment, cognitive impairment, and / or gait deterioration associated with Taysaks disease. For example, acetyl-leucine or a pharmaceutically acceptable salt thereof may delay the onset of decreased coordination, tremor, motor impairment, cognitive impairment, and / or gait deterioration associated with Taysaks disease, reduce its severity or eliminate it, or delay or reverse its progression.

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

[0104] The "therapeutic effective amount" of a drug is any amount that, when administered to a subject, is the amount of drug required to produce the desired effect, which may be therapeutic and / or prophylactic in the context of this disclosure. Dosage can be determined based on various parameters, such as the specific 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. The physician will be able to determine the required route of administration and dosage for any given patient. For example, a daily dose may be about 10 to about 225 mg / kg, about 10 to about 150 mg / kg, or about 10 to about 100 mg / kg body weight.

[0105] A kit for treating LSD in subjects in need (e.g., subjects with, suspected of having, or at risk of having LSD) is also disclosed, comprising a device for diagnosing or predicting LSD, and acetyl-leucine or a pharmaceutically acceptable salt thereof.

[0106] Devices for diagnosing or predicting LSD may include specific binders, probes, primers, primer pairs or primer combinations, enzymes, or antibodies, wherein the antibody includes antibody fragments capable of detecting or aiding in the detection of LSD as defined herein. Kits may contain LysoTracker. ® It is a fluorescent marker and is commercially available from Invitrogen and Lonza. LysoTracker® can be blue, blue-white, yellow, green, or red.

[0107] The kit also contains acetyl-leucine or a pharmaceutically acceptable salt thereof as defined herein. The kit may further contain a buffer or aqueous solution. The kit may further include instructions for using acetyl-leucine or a pharmaceutically acceptable salt thereof in the methods of the present invention.

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

[0109] As used herein, “neuroprotection” and its synonyms refer to the prevention, slowing, and / or reversal of neurodegeneration, including but not limited to progressive loss of neuronal structure, progressive loss of neuronal function, and / or progressive neuronal death. Providing neuroprotection may result in delayed onset of LSD or one or more symptoms of LSD (the disease or symptoms that would otherwise be expected to develop according to typical disease progression), reduced severity of LSD, or reduced or eliminated severity of one or more existing symptoms associated with LSD, delayed or reversed the progression of LSD or one or more symptoms of LSD over time compared to typical disease progression, and / or reversed the progression of LSD or one or more symptoms of LSD over time. The duration of neuroprotection may coincide with the duration of treatment described herein. Furthermore, for example, treatment may provide neuroprotection for a duration of, for example, more than 7 days, more than 2 weeks, more than 3 weeks, more than 1 month, more than 6 weeks, more than 7 weeks, or more than 2 months. Treatment may provide neuroprotection for a duration of more than 3 months, more than 4 months, more than 5 months, or more than 6 months. In another embodiment, it provides neuroprotection for a duration of, for example, more than one year, more than two years, more than three years, more than four years, more than five years, or more than ten years. The treatment provides neuroprotection throughout the patient's life.

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

[0111] In one embodiment, there is a method for providing neuroprotection in subjects who have, are suspected of having, or are 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 subjects who have, are suspected of having, or are at risk of having LSD, the kit comprising a device for diagnosing or predicting LSD, and acetyl-leucine or a pharmaceutically acceptable salt thereof.

[0113] This disclosure also includes the use of acetyl-leucine or a pharmaceutically acceptable salt thereof as a neuroprotective agent in subjects who have, are suspected of having, or are at risk of having LSD.

[0114] All features described herein (including any appended claims, abstract, and drawings) and / or all steps of any method so disclosed may be combined with any of the foregoing aspects in any combination, except that at least some of the described features and / or steps are mutually exclusive combinations.

[0115] Example

[0116] The invention will now be explained in further detail in the following examples, which demonstrate the role of acetyl-leucine in treating LSD in subjects and providing neuroprotection in said subjects.

[0117] Example 1

[0118] In vivo mouse studies - methods

[0119] mouse model

[0120] This study utilized a real mouse model of NPC, namely Npc1 - / - (BALB / cNctr- Npc1 m1N / J Mice that were ineffective against the NPC1 protein and showed all the hallmarks of clinical disease (Loftus, 1997).

[0121] This mutation arises spontaneously and has a lifespan of 10-14 weeks, thus resulting in a more acute disease course in the vast majority of patients. Mutant mice have been successfully developed not only to determine the individual occurrence and potential pathogenic mechanisms of the disease but also to evaluate experimental therapies. Analysis using these mice has been conducted 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 about 4-5 weeks of age, Npc1 - / - The mice did not exhibit any obvious behavioral signs of disease that distinguished them from their wild-type littermates. The first signs of behavioral deficits, such as tremors and ataxia, appeared at week 5–6; by week 7–8, the deficits in motor coordination became more pronounced, and by week 9–10, ataxia progressed, accompanied by worsening weight loss and poor coat condition due to difficulty feeding and drinking (applied humane endpoints) (Smith, 2009).

[0123] wild type ( Npc1 - / - Mice from the same littermate were used as controls.

[0124] Treatment Procedure

[0125] From weaning (3 weeks old), Npc1 - / - mouse group and Npc1 + / + The mouse group was treated with 0.1 g / kg acetyl-DL-leucine, which was provided in the mouse diet. Alone Npc1 - / - and Npc1 + / + The mouse group was used as a control group for treatment.

[0126] fur condition

[0127] Through a simple observation of 9-week-old mice, we compared mice that had received acetyl-DL-leucine treatment with those that had not. Npc1 - / - The condition of the mouse's fur.

[0128] Weight data

[0129] The animals were weighed twice a week. The average weight of all mice in each group was calculated and compared.

[0130] Gait analysis

[0131] According to the manufacturer's instructions (Noldus, Nottingham, UK), use CatWalk. ® The 15.0 system was used to analyze the gait of 8-week-old mice. Five runs were recorded for each animal.

[0132] The measured CatWalk® parameters are as follows: 1. Average standing time: The average duration (s) of contact between the claw and the glass plate; 2. Step cycle: The duration (in seconds) between two consecutive contacts of the same claw; 3. Duty cycle: The percentage of time the claw is in contact with the plate compared to the time it takes to complete a step cycle; 4. Step sequence (AB): Percentage of time spent walking in the alternating LF-RH-RF-LH pattern (LF: left front; RH: right back; RF: right front; LH: left back); 5. Step frequency: The number of steps per second during the experiment; 6. Diagonal support: Percentage of time during which the diagonal claws are in simultaneous contact with the glass plate (RF&LH or RH&LF).

[0133] Motor function analysis

[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] fur condition

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

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

[0140] Weight data

[0141] from Figure 2A It can be seen from the wild type ( Npc1 + / + The mice gradually gained weight during the study period, from 3 to 10 weeks of age. Furthermore, Figure 2A Showing the average body weight of each group of mice at each time point ( Npc1 - / - For untreated cases, n = 1; Npc1 - / - Acetyl-DL-leucine 0.1 g / kg, n = 3; Npc1 + / + For untreated cases, 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 - / - The mice initially gained weight, primarily due to... Npc1 + / + The control group followed the same method. However, then... Npc1 - / - The mice began losing weight at six weeks of age. By the end of the study (10 weeks old), the mice weighed almost as little as they did at four weeks of age.

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

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

[0146] Gait analysis

[0147] The results of the gait analysis are shown in Figure 3. Diagonal support, gait frequency, and gait sequence data are shown in the figures below. Figures 3A-3C middle. Figure 3D and 3E Displayed forepaw (FP) data ( Figure 3D Average standing time and gait cycle; Figure 3E (duty cycle in the middle). Figure 3F and 3G Displayed hind paw (HP) data ( Figure 3FAverage standing time and gait cycle; Figure 3G (Duty cycle in the data). Data are presented as mean ± SEM. For untreated... Npc1 + / + n=3, for treatment Npc1 + / + n=2, for untreated Npc1 - / - n=1 (therefore no statistical analysis was performed), for treatment Npc1 - / - n=3.

[0148] The first bar in each chart shows the wild type ( Npc1 + / + Gait characteristics of mice.

[0149] The second bar in each chart shows the wild-type treated with acetyl-DL-leucine ( Npc1 + / + The gait characteristics of these mice were not significantly different from those of their untreated littermates.

[0150] The third bar in each chart shows Npc1 - / - Gait characteristics of mice. Overall, compared with... Npc1 + / + Compared to other mice, this mouse exhibited a poorer gait. The mouse showed poorer gait in diagonal support (…). Figure 3A ) or step sequence ( Figure 3C It takes very little time (if any) in the process, and its hind paw function ( Figure 3F ) and duty cycle ( Figure 3G It has also been severely hampered.

[0151] The fourth bar in each chart shows treatment with acetyl-DL-leucine. Npc1 - / - Gait characteristics of the mice. Compared with untreated littermates, these mice exhibited a significantly improved gait. In fact, they showed a gait similar to that of their littermates. Npc1 + / + Similar gait characteristics to mice.

[0152] Motor function analysis

[0153] Analysis at 8 weeks of age showed Npc1 - / - and wild type ( Npc1 + / + There were no differences in motor function characteristics among the mice (data not shown).

[0154] However, by the time the child was 9 weeks old, the deficit in motor coordination had become apparent.

[0155] The results of motor function analysis at 9 weeks are shown in Figure 4. Central upright, active, upright, and front-to-back (FR) counts are displayed respectively. Figures 4A-4D The activity time, movement time, standing time, and total manual standing count are displayed separately. Figure 4E-4H Data are presented as mean ± SEM. For untreated... Npc1 + / + n=3, for treatment Npc1 + / + n=2, for untreated Npc1 - / - n=1 (therefore no statistical analysis was performed), for treatment Npc1 - / - ， n=3.

[0156] The first bar in each chart shows the wild type ( Npc1 + / + Motor function characteristics of mice.

[0157] The second bar in each chart shows the wild-type treated with acetyl-DL-leucine ( Npc1 + / + Motor function characteristics of mice. There were no significant differences in motor function characteristics between these mice and their untreated littermates.

[0158] The third bar in each chart shows Npc1 - / - Motor function characteristics of mice. Overall, compared with... Npc1 + / + Compared to mice, this mouse exhibited poorer motor function. The mouse spent very little time (if any) standing upright (Fig. H), especially standing upright on its hind legs without support (Fig. A).

[0159] The fourth bar in each chart shows treatment with acetyl-DL-leucine. Npc1 - / - Motor function characteristics of the mice. Compared with their untreated littermates, these mice exhibited significantly improved motor function. In fact, they showed similar characteristics to their littermates. Npc1 + / + Similar motor function characteristics to mice.

[0160] life

[0161] It was also observed that treatment with acetyl-DL-leucine (0.1 g / kg starting at 3 weeks of age) resulted in Npc1 - / - Mice showed a statistically significant association with increased lifespan. Figure 5 This data further demonstrates the role of acetyl-leucine in delaying the onset of disease.

[0162] in conclusion

[0163] exist Npc1 - / - Mice exhibiting clear signs of disease from 5-6 weeks of age, distinguishing them from wild-type littermates, were treated with acetyl-DL-leucine from weaning. Npc1 - / - The pups in the same litter did not show these symptoms until more than two weeks later. Treatment with acetyl-DL-leucine was administered. Npc1 - / - The mice showed delayed onset and progression of NPC symptoms and evidence of neuroprotective effects.

[0164] Example 2

[0165] method

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

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

[0168] result

[0169] Treatment of fibroblasts derived from patients with mild clinical severity of NPC with 1 mM N-acetyl-DL-leucine was significantly associated with a decrease in LysoTracker fluorescence, indicating a reduction in lysosomal volume over time. Figure 6A These findings were replicated in fibroblasts from other NPC patients with variable clinical severity, which were treated with 1 mM N-acetyl-DL-leucine for 72 hours. Figure 6B ).

[0170] Fibroblasts from patients with NPA, MLII, MPS IIIB, aspartate glucosamineuria, MLIIIA, and MPS VII were observed to have elevated LysoTracker fluorescence levels compared to age-matched wild-type controls. Figure 6C-6H This indicates that lysosomal enlargement occurs due to lipid accumulation compared to fibroblasts from healthy individuals. Acetyl-leucine treatment was statistically significantly associated with a decrease in LysoTracker fluorescence relative to control levels in NPA, MLII, and MPS IIIB fibroblasts compared to untreated NPA, MLII, and MPS IIIB fibroblasts. Figure 6C-6E Furthermore, treatment with acetyl-leucine was associated with a decreasing trend in LysoTracker fluorescence relative to control levels in aspartic glucosamineuria, MLIIIA, and MPS VII fibroblasts, respectively. Figure 6F-6H The decrease in LysoTracker fluorescence indicates a reduction in lysozyme volume. Figure 6C-6H ). Figures 6A-6D The data shown represent the treatment outcomes for each cell line, with lysosomal volume expressed as a fold change relative to untreated wild-type fibroblasts. (Astro) / The p-value represents the difference between the treated fibroblasts and untreated fibroblasts (<0.05 / 0.001).

[0171] in conclusion

[0172] N-acetyl-DL-leucine treatment corrects disrupted lysosomal storage by reducing lysosome volume, thus directly correcting the phenotype of these lysosomal storage disorders. These disorders represent different types of LSD, therefore these results further support the role of acetyl-leucine in a wide range of lysosomal storage disorders.

[0173] Example 3

[0174] Sandhoff's disease is an autosomal recessive disorder likely caused by mutations in the HEXB gene, which encodes the β-subunit of β-aminohexosidase. As a result, GM2 gangliosides cannot be degraded and accumulate in lysosomes in peripheral and central nervous system (CNS) cells.

[0175] This study used a mouse model of Sandhoff's disease. Hexb - / -Mice, as described by Jeyakumar et al. (Jeyakumar, M. et al. (1999) Proc. Natl. Acad. Sci. USA 96: 6388-6393).

[0176] wild type ( Hexb + / + Mice were used as controls.

[0177] life

[0178] Treatment with acetyl-DL-leucine resulted in a statistically significant increase in lifespan in Sandhoff mice. Figure 7A (Related to) Figure 7A 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] Bar test

[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 7B ).exist Figure 7BIn this study, acetyl-leucine was administered at a dose of 0.1 g / kg body weight starting at 3 weeks of age. Acetyl-leucine-treated Sandhoff mice retained the ability to complete the test until 13 weeks of age (inclusive). The data shown are the mean of 6 mice per group. Treated Sandhoff mice retained the ability to complete the test until 13 weeks of age (inclusive).

[0185] Step cycle

[0186] The stride cycle is the time required for a limb to move from the moment it leaves the ground until the next time it leaves the ground.

[0187] The gait cycle time was assessed 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 of acetyl-leucine starting from 3 weeks of age.

[0188] Treatment of the Sandhoff mouse model with acetyl-leucine was associated with significantly faster pre-step cycle time (p < 0.05 vs untreated SH mice), significantly faster post-step cycle time (p < 0.01 vs untreated SH mice), and significantly faster mean step cycle time (p < 0.001 vs untreated SH mice). Figure 7C ).exist Figure 7C In this study, mice were treated with acetyl-leucine at a dose of 0.1 g / kg body weight starting at 3 weeks of age. The pre-gait cycle refers to the forelimbs, and the post-mean gait cycle refers to the hindlimbs; the mean gait cycle considers all four limbs of the mice. (Astro) / / () indicates a p-value < 0.05 / 0.01 / 0.001 relative to untreated Sandhoff mice. The data shown are mean ± Stdev.

[0189] Therefore, in the Sandhoff mouse model, acetyl-leucine treatment was associated with a faster gait cycle, which may indicate improved motor function.

[0190] in conclusion

[0191] These studies demonstrate that acetyl-leucine treatment in a mouse model of Sandhoff's disease improved motor function and significantly extended lifespan, as assessed in two independent experiments.

[0192] Example 4

[0193] GM2 ganglioside storage disorders are a group of lysosomal storage disorders caused by a deficiency in β-aminohexosidase activity. This group includes Tessachus disease, Sandhoff's disease, and the AB variant of Tessachus disease.

[0194] Fibroblasts from GM2 patients (Tessachus disease, Sandhoff disease, and the AB variant of Tessachus disease) and healthy controls were treated with acetyl-DL-leucine (1 mM for 6 days), followed by extraction and quantification of glycosphingolipid (GSL) levels by high-performance liquid chromatography (HPLC).

[0195] In the absence of treatment, fibroblasts from all three strains of GM2 ganglioside storage disease 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. This decrease was statistically significant in the case of Tessack disease (p<0.05). A trend toward treatment-related decreases in GSL levels was observed in the cases of Sandhof disease and the AB variant of Tessack disease. Figures 8A-8C The data shown represent the treatment outcomes for each cell line, where GSL levels are adjusted relative to protein content and expressed as fold changes relative to levels in untreated wild-type fibroblasts.

[0196] Example 5

[0197] Patient 1

[0198] The patient in this study was a 28-year-old male genetically diagnosed with Taysaks disease, exhibiting dysarthria, tremor, ataxia of posture and gait, hemiparesis of the lower limbs, and muscle atrophy. Specifically, the patient was unable to stand or walk, could take a single step with strong support, and displayed significant postural instability, eye movement disorders, dysphagia, dysarthria, and mild cognitive impairment. The first symptoms were observed at age 16.

[0199] Prior to treatment, the patient's examination showed an ataxia assessment rating of 15.5 / 40 on the Special Ataxia Assessment Scale (SARA). Furthermore, the analysis results of the patient's spinocerebellar ataxia functional index (SCAFI) are as follows: Average 8-meter walking test (8MW): 21.6 s MW 9-hole socket test advantages (9HPTD) (right side): 48.3 s MW 9-hole insert test non-dominant (9HPTND): 44.9 s MW PATA Vocabulary Test: 20 Montreal Cognitive Assessment (MoCA): 18 / 30 They also recorded videos of the patients for later comparison.

[0200] The day after the examination, the patient began treatment with acetyl-leucine, with a dose of 3 g / day for the first week, followed by a dose of 5 g / day for the second week and thereafter.

[0201] The patient was re-examined at 1 month and 4 months, while continuing treatment. At 1 month, the patient had improved fine motor skills and reduced hand tremors, such as when eating or drinking. Walking remained largely unchanged. At 4 months, the patient was stable with slight improvement in cognitive function, but postural, gait, and motor function had all declined. The patient's SARA score and SCAFI analysis results compared to baseline are shown below.

[0202] Table 1. Patient assessment parameters.

[0203]

[0204] Overall, patients showed symptom improvement after treatment with acetyl-leucine.

[0205] Patient 2

[0206] The patient in this study was a 32-year-old woman genetically diagnosed with Taysaks disease, exhibiting combined postural and gait incoordination, fine motor impairment, hemiparesis of the lower limbs, and muscle atrophy. Specifically, she was unable to walk without support, and also suffered from dysphagia, speech impairment, oculomotor dysfunction, and mild cognitive impairment. The first symptoms were observed at age 7.

[0207] Prior to treatment, the patient's examination showed an ataxia assessment rating of 10.5 / 40 on the Special Ataxia Assessment Scale (SARA). Furthermore, the analysis results of the patient's spinocerebellar ataxia functional index (SCAFI) are as follows: Average 8-meter walking test (8MW): 12.5 s MW 9-hole socket test advantage (9HPTD) (right side): 21.5 s MW 9-hole insert test non-dominant (9HPTND): 35.5 s MW PATA Vocabulary Test: 18 Montreal Cognitive Assessment (MoCA): 21 / 30 They also recorded videos of the patients for later comparison.

[0208] On the day of the examination, the patient began treatment with acetyl-leucine, with a dose of 3 g / day for the first week, and then 5 g / day for the second week and thereafter.

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

[0210] Table 2. Patient assessment parameters.

[0211] Patient 3

[0212] The patient in this study was an 8-year-old male genetically diagnosed with Taysaks disease. He experienced epileptic seizures (tonic-clonic, approximately 10 seconds, self-limiting) almost daily before falling asleep, along with oculomotor disturbances, dysarthria, and significant cognitive and attentional problems (not apparent on neurological examination). He was unable to stand or walk independently and was severely limited in his daily activities (unable to eat, wash, or dress himself). The first symptom was observed at 9 months of age.

[0213] Prior to treatment, the patient's examination showed an ataxia assessment rating of 36 / 40, a mRDS score of 18 / 24, an EQ-5D-5L visual scale score of 50, and an 8MWT score of 18.1 (with strong support only).

[0214] 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.

[0215] One month later, the patient underwent a follow-up examination while continuing treatment and showed increased fine motor skills (ability to grasp small objects), increased ambition (more attempts to walk independently), improved postural stability, gait, and posture, and the ability to speak 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 study was a 13-year-old male who was genetically diagnosed with GM1 ganglioside storage disease. He was unable to stand or walk independently, had severely limited daily activities (unable to eat, wash, or dress himself), and exhibited oculomotor and dysarthria symptoms. He also had significant cognitive and attentional problems (which were not apparent on neurological examination). The first symptoms were observed at age 2.

[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,” Am J Med Genet Part B 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 patients with NPC received a long-term course of acetyl-leucine at 4.5 g / day. Core seizure response (CSS) was determined at baseline and at various time points for eye movement, gait, language, swallowing, fine motor skills, cognition, memory, and seizures. Overall CSS was calculated at baseline and at each such time point by adding a separate CSS value for each parameter (eye movement, gait, etc.). The number of days after treatment initiation for CSS assessment varied for each patient, as shown in Table 5.

[0226] Table 5. Number of days after initiation of acetyl-leucine treatment for CSS.

[0227]

[0228] Tables 6-14 below show the CSS for general, eye movement, walking, language, swallowing, fine motor skills, cognition, memory, and seizures, respectively.

[0229] Table 6. Overall CSS.

[0230]

[0231] Table 7. Eye Movement CSS.

[0232]

[0233] Table 8. CSS for walking around.

[0234]

[0235] Table 9. Language CSS.

[0236]

[0237] Table 10. Swallowing CSS.

[0238]

[0239] Table 11. Fine motor skills CSS.

[0240]

[0241] Table 12. Understanding CSS.

[0242]

[0243] Table 13. Memory CSS.

[0244]

[0245] Table 14. Seizure CSS.

[0246]

[0247] Baseline and ASIS at each time point were calculated using each patient's CSS and age at assessment. 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 Table 6 and Figure 9A As shown, none of the 10 patients exhibited an overall increase in CSS during 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 constant CSS during 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 during the experiment, indicating that the disease did not progress and was actually significantly reduced. Improvements were observed in different subscores: Patient 1: walking; Patient 3: fine motor skills; Patient 4: walking and language; Patient 8: eye movements and fine motor skills; Patient 9: memory; Patient 10: cognition. Figure 10A-10J The data presented here show the CSS sub-scores for each patient, presented in the form of a bar chart.

[0251] As shown in Table 15 and Figure 9B As shown, all 10 patients exhibited a reduction in ASIS relative to baseline during treatment. In patients 2, 5, 6, and 7, CSS remained the same with increasing age, resulting in a small reduction in ASIS. In patients 1, 3, 4, 8, 9, and 10, the reduction in ASIS was greater due to a decrease in CSS with increasing age.

Claims

1. The use of acetyl-leucine or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating lysosomal storage disease (LSD) or one or more symptoms associated with LSD in a subject in need, wherein the LSD is selected from Niemann-Pick disease type C (NPC), Tessachs disease, AB variant of Tessachs disease, Sandhof disease, Niemann-Pick disease type A, mucosal storage disease type II, mucosal storage disease type III, MPS III, MPS VII, GM1 ganglioside storage disease, aspartic glucosamineuria, Niemann-Pick disease type B, Fabry disease, neuronal ceroid lipofuscin storage disease (NCL), Barten disease, Kuff's disease, palmitoyl thioesterase-1 deficiency (type A), cathepsin F deficiency (type B), Krabby disease, Fabry disease, Gaucher disease, metachromatic leukodystrophy, multiple sulfatase deficiency, Sheldr's disease, Morquio Disease B, GM2 ganglioside storage disease, lysosomal acid lipase deficiency, galacturonic acidosis, type I mucolipid storage disease, type IV mucolipid storage disease, MPS IH, MPS IHS, MPS IS, MPS IIA, MPS IIB, MPS IVA, MPS VI, MPS IX, β-mannosinoside storage disease, α-fucoside storage disease, Niemann-Pick disease type D, Wollman's disease, Danon's disease, cystinosis, osteogenesis imperfecta condensans, sialic acid storage disease, and infantile free sialic acid storage disease, wherein the ataxia assessment rating scale (SARA) scores of the subjects described above improved compared to pre-drug administration.

2. The application as described in claim 1, wherein the LSD is selected from Tessachus disease, the AB variant of Tessachus disease, Sandhof disease, and GM1 ganglioside storage disease.

3. The application as described in claim 1 or 2, wherein the application comprises initially administering a therapeutically effective amount of acetyl-leucine to a subject in need while the subject is asymptomatic.

4. The application as described in claim 3, wherein the initial administration is performed after the subject is found to have the genetic and / or biochemical markers of the LSD.

5. The application as described in claim 1 or 2, wherein the application comprises administering a therapeutically effective amount of acetyl-leucine to a subject in need for a duration selected from at least 3 months, at least 6 months, at least 1 year, at least 2 years, and at least 5 years.

6. The application as described in claim 1 or 2, wherein the acetyl-leucine is acetyl-DL-leucine.

7. The application as described in claim 1 or 2, wherein the acetyl-leucine has an enantiomer excess of the L-enantiomer or the D-enantiomer.

8. The application as described in claim 1 or 2, wherein the acetyl-leucine is a single enantiomer of the L-enantiomer or the D-enantiomer.

9. The application as described in claim 8, wherein the single enantiomer form is an L-enantiomer.

10. The application as described in claim 1 or 2, wherein the application comprises administering to a subject in need a therapeutically effective amount of acetyl-leucine at a dose of 1 g to 15 g / day, 1 g to 10 g / day, 1.5 g to 7 g / day, 4 g to 6 g / day, or 4 g to 5 g / day.

11. The use of acetyl-leucine or a pharmaceutically acceptable salt thereof in the preparation of a medicament for delaying the onset of LSD or one or more symptoms of LSD, which would otherwise be expected to occur according to typical disease progression, wherein the LSD is selected from Niemann-Pick disease type C (NPC), Tessachs disease, AB variant of Tessachs disease, Sandhof disease, Niemann-Pick disease type A, mucolipid storage disease type II, mucolipid storage disease type III, MPS III, MPS VII, GM1 ganglioside storage disease, aspartate glucosamineuria, Niemann-Pick disease type B, Fabry disease, neuronal ceroid lipofuscin storage disease (NCL), Barten disease, Kuff's disease, palmitoyl thioesterase-1 deficiency (type A), cathepsin F deficiency (type B), Krabby disease, Fabry disease, Gaucher disease, metachromatic leukodystrophy, multiple sulfatase deficiency, Sheldr's disease, Morquio Disease B, GM2 ganglioside storage disease, lysosomal acid lipase deficiency, galacturonic acidosis, type I mucolipid storage disease, type IV mucolipid storage disease, MPS IH, MPS IH-S, MPS IS, MPS IIA, MPS IIB, MPS IVA, MPS VI, MPS IX, β-mannosinoside storage disease, α-fucoside storage disease, Niemann-Pick disease type D, Wollman's disease, Danon's disease, cystinosis, osteogenesis imperfecta condensans, sialic acid storage disease, and infantile free sialic acid storage disease, wherein the ataxia assessment rating scale (SARA) scores of the subjects described above improved compared to pre-drug administration.

12. The use of acetyl-leucine or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating lysosomal storage disease (LSD) or one or more symptoms associated with LSD in a subject in need, wherein the use comprises administering a therapeutically effective amount of acetyl-leucine to the subject in need for a duration selected from 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 Niemann-Pick disease type C (NPC), Tessachs disease, AB variant of Tessachs disease, Sandhof disease, Niemann-Pick disease type A, mucosal storage disease type II, mucosal storage disease type III, MPS III, MPS VII. GM1 ganglioside storage disease, aspartic glucosamineuria, Niemann-Pick disease type B, Fabry disease, neuronal ceroid lipofuscin storage disease (NCL), Barten disease, Kuff's disease, palmitoylprotein thioesterase-1 deficiency (type A), cathepsin F deficiency (type B), Krabby's disease, Fabry's disease, Gaucher's disease, metachromatic leukodystrophy, multiple sulfatase deficiencies, Sheldr's disease, Morquio B disease, GM2 ganglioside storage disease, lysosomal acid lipase deficiency, galacturonic acidosis, type I mucolipid storage disease, type IV mucolipid storage disease, MPS IH, MPS IHS, MPS IS, MPS IIA, MPS IIB, MPS IVA, MPS VI, MPS IX, β-mannosinolate storage disease, α-fucoside storage disease, Niemann-Pick disease type D, Wollman's disease, Danon's disease, cystinosis, osteogenesis imperfecta condensans, sialic acid storage disease, and infantile free sialic acid storage disease, wherein the ataxia assessment rating scale (SARA) scores of the subjects described above improved compared to pre-drug administration.

13. The application as described in claim 12, wherein the therapeutically effective amount of acetyl-leucine is 1 g to 15 g / day, 1 g to 10 g / day, 1.5 g to 7 g / day, 4 g to 6 g / day, or 4 g to 5 g / day.

14. The use of acetyl-leucine or a pharmaceutically acceptable salt thereof in the preparation of a medicament for delaying the progression of LSD or one or more LSD-related symptoms over time compared with typical disease progression, wherein said use comprises administering a therapeutically effective amount of acetyl-leucine to a subject in need for a duration selected from at least 3 months, at least 6 months, at least 1 year, at least 2 years, and at least 5 years, and wherein said LSD is selected from Niemann-Pick disease type C (NPC), Tessachs disease, AB variant of Tessachs disease, Sandhof disease, Niemann-Pick disease type A, Mucosal storage disease type II, Mucosal storage disease type III, MPS III, MPS VII. GM1 ganglioside storage disease, aspartic glucosamineuria, Niemann-Pick disease type B, Fabry disease, neuronal ceroid lipofuscin storage disease (NCL), Barten disease, Kuff's disease, palmitoylprotein thioesterase-1 deficiency (type A), cathepsin F deficiency (type B), Krabby's disease, Fabry's disease, Gaucher's disease, metachromatic leukodystrophy, multiple sulfatase deficiencies, Sheldr's disease, Morquio B disease, GM2 ganglioside storage disease, lysosomal acid lipase deficiency, galacturonic acidosis, type I mucolipid storage disease, type IV mucolipid storage disease, MPS IH, MPS IHS, MPS IS, MPS IIA, MPS IIB, MPS IVA, MPS VI, MPS IX, β-mannosinolate storage disease, α-fucoside storage disease, Niemann-Pick disease type D, Wollman's disease, Danon's disease, cystinosis, osteogenesis imperfecta condensans, sialic acid storage disease, and infantile free sialic acid storage disease, wherein the ataxia assessment rating scale (SARA) scores of the subjects described above improved compared to pre-drug administration.

15. The use of acetyl-leucine or a pharmaceutically acceptable salt thereof in the preparation of a medicament for reversing the progression of LSD or one or more LSD-related symptoms over time, wherein the use comprises administering a therapeutically effective amount of acetyl-leucine to a subject in need for a duration selected from 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 Niemann-Pick disease type C (NPC), Tessachs disease, AB variant of Tessachs disease, Sandhof disease, Niemann-Pick disease type A, mucolipidosis type II, mucolipidosis type III, MPS III, MPS VII. GM1 ganglioside storage disease, aspartic glucosamineuria, Niemann-Pick disease type B, Fabry disease, neuronal ceroid lipofuscin storage disease (NCL), Barten disease, Kuff's disease, palmitoyl thioesterase-1 deficiency (type A), cathepsin F deficiency (type B), Krabby's disease, Fabry's disease, Gaucher's disease, metachromatic leukodystrophy, multiple sulfatase deficiencies, Sheldr's disease, Morquio B disease, GM2 ganglioside storage disease, lysosomal acid lipase deficiency, galacturonic acidosis, type I mucolipid storage disease, type IV mucolipid storage disease, MPS IH, MPS IHS, MPS IS, MPS IIA, MPS IIB, MPS IVA, MPS VI, MPS IX, β-mannosinolate storage disease, α-fucoside storage disease, Niemann-Pick disease type D, Wollman's disease, Danon's disease, cystinosis, osteogenesis imperfecta condensans, sialic acid storage disease, and infantile free sialic acid storage disease, wherein the ataxia assessment rating scale (SARA) scores of the subjects described above improved compared to pre-drug administration.

16. The use of acetyl-leucine or a pharmaceutically acceptable salt thereof in the preparation of a medicament for improving a biochemical marker of LSD in a subject in need over time, wherein said use comprises administering a therapeutically effective amount of acetyl-leucine to a subject in need for a duration selected from at least 3 months, at least 6 months, at least 1 year, at least 2 years, and at least 5 years, and wherein said LSD is selected from Niemann-Pick disease type C (NPC), Tessachs disease, AB variant of Tessachs disease, Sandhof disease, Niemann-Pick disease type A, mucosal storage disease type II, mucosal storage disease type III, MPS III, MPS VII. GM1 ganglioside storage disease, aspartic glucosamineuria, Niemann-Pick disease type B, Fabry disease, neuronal ceroid lipofuscin storage disease (NCL), Barten disease, Kuff's disease, palmitoyl thioesterase-1 deficiency (type A), cathepsin F deficiency (type B), Krabby's disease, Fabry's disease, Gaucher's disease, metachromatic leukodystrophy, multiple sulfatase deficiencies, Sheldr's disease, Morquio B disease, GM2 ganglioside storage disease, lysosomal acid lipase deficiency, galacturonic acidosis, type I mucolipid storage disease, type IV mucolipid storage disease, MPS IH, MPS IHS, MPS IS, MPS IIA, MPS IIB, MPS IVA, MPS VI, MPS IX, β-mannosinolate storage disease, α-fucoside storage disease, Niemann-Pick disease type D, Wollman's disease, Danon's disease, cystinosis, osteogenesis imperfecta condensans, sialic acid storage disease, and infantile free sialic acid storage disease, wherein the ataxia assessment rating scale (SARA) scores of the subjects described above improved compared to pre-drug administration.

17. The application of claim 16, wherein the biochemical marker is an increased lysosomal volume.

18. The use of acetyl-leucine or a pharmaceutically acceptable salt thereof in the preparation of a medicament for reducing the severity of LSD, or reducing the severity of one or more existing symptoms associated with LSD, or eliminating one or more existing symptoms associated with LSD in subjects in need, wherein the LSD is selected from Niemann-Pick disease type C (NPC), Tessachs disease, AB variant of Tessachs disease, Sandhof disease, Niemann-Pick disease type A, mucolipid storage disease type II, mucolipid storage disease type III, MPS III, MPS VII, GM1 ganglioside storage disease, aspartate glucosamineuria, Niemann-Pick disease type B, Fabry disease, neuronal ceroid lipofuscin storage disease (NCL), Barten disease, Kuff's disease, palmitoyl thioesterase-1 deficiency (type A), cathepsin F deficiency (type B), Krabby disease, Fabry disease, Gaucher disease, metachromatic leukodystrophy, multiple sulfatase deficiency, Sheldr's disease, Morquio Disease B, GM2 ganglioside storage disease, lysosomal acid lipase deficiency, galacturonic acidosis, type I mucolipid storage disease, type IV mucolipid storage disease, MPS IH, MPS IHS, MPS IS, MPS IIA, MPS IIB, MPS IVA, MPS VI, MPS IX, β-mannosinoside storage disease, α-fucoside storage disease, Niemann-Pick disease type D, Wollman's disease, Danon's disease, cystinosis, osteogenesis imperfecta condensans, sialic acid storage disease, and infantile free sialic acid storage disease, wherein the ataxia assessment rating scale (SARA) scores of the subjects described above improved compared to pre-drug administration.

19. The use of acetyl-leucine or a pharmaceutically acceptable salt thereof in the preparation of a medicament for providing neuroprotection in subjects who have, are suspected of having, or are at risk of having LSD, wherein said use comprises administering a therapeutically effective amount of acetyl-leucine to a subject in need for a duration selected from at least 3 months, at least 6 months, at least 1 year, at least 2 years, and at least 5 years, and wherein said LSD is selected from Niemann-Pick disease type C (NPC), Tessachs disease, AB variant of Tessachs disease, Sandhof disease, Niemann-Pick disease type A, mucolipidosis type II, mucolipidosis type III, MPS III, MPS VII. GM1 ganglioside storage disease, aspartic glucosamineuria, Niemann-Pick disease type B, Fabry disease, neuronal ceroid lipofuscin storage disease (NCL), Barten disease, Kuff's disease, palmitoyl thioesterase-1 deficiency (type A), cathepsin F deficiency (type B), Krabby's disease, Fabry's disease, Gaucher's disease, metachromatic leukodystrophy, multiple sulfatase deficiencies, Sheldr's disease, Morquio B disease, GM2 ganglioside storage disease, lysosomal acid lipase deficiency, galacturonic acidosis, type I mucolipid storage disease, type IV mucolipid storage disease, MPS IH, MPS IHS, MPS IS, MPS IIA, MPS IIB, MPS IVA, MPS VI, MPS IX, β-mannosinolate storage disease, α-fucoside storage disease, Niemann-Pick disease type D, Wollman's disease, Danon's disease, cystinosis, osteogenesis imperfecta condensans, sialic acid storage disease, and infantile free sialic acid storage disease, wherein the ataxia assessment rating scale (SARA) scores of the subjects described above improved compared to pre-drug administration.

20. The application according to any one of claims 1 to 19, wherein the subject's SARA score has improved by at least one point compared to before administration.

21. The application according to any one of claims 1 to 20, wherein the subject achieved improvement in one or more of the following assessments compared to pre-drug administration: Functional SARA (fSARA), Spinocerebellar Ataxia Functional Index (SCAFI), Modified Disability Rating Scale (mDRS), Niemann-Pick Disease Type C Clinical Severity Scale (NPC-CSS), and EuroQol 5-Dimensional 5-Level (EQ-5D-5L) assessment questionnaire.

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