Modified forms of ambroxol for therapeutic use

Abstract

The present invention relates to analog forms of ambroxol and related compounds, compositions comprising them, and methods for preventing and / or treating various diseases and medical conditions with the administration of analogs of ambroxol and related compounds. The present invention also provides pharmaceutical compositions comprising a compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof), and methods for preventing and / or treating various diseases and medical conditions in a subject with the administration of a compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof).

Description

[Technical Field]

[0001] Background of the Invention Field of Invention The present invention relates to derivatives or analogs of ambroxol, ambroxol hydrochloride, and / or bromhexine, which are suitable for use in various therapeutic applications. [Background technology]

[0002] Consideration of related technologies Ambroxol, ambroxol hydrochloride, and its structurally related “parent compound” bromhexine are mucolytics that have been available in several countries since the late 1970s for use in the treatment of acute and chronic respiratory diseases and conditions associated with the production of excessive and / or highly viscous mucus.

[0003] By inducing lysosomal exocytosis via pH-dependent calcium release from acidic calcium reservoirs (Fois G et al., Cell Calcium 58(6):628-637, 2015), ambroxol is thought to reduce mucus adhesion to the airway walls by promoting mucus clearance, for example, by breaking down sputum and stimulating surfactant production by type II alveolar epithelial cells (Seiffert C et al., Toxicol Appl Pharmacol 203(1):27-35, 2005). In addition, ambroxol has been shown to enable rapid pain relief used in acute pharyngitis by potently inhibiting nerve sodium channels, particularly when administered in candy form (de Mey C et al., Arzneimittel-Forschung 28(5a):889-898, 1978).

[0004] More recently, there has been broad interest in the repurposed use of ambroxol for several other medical applications. For example, it has been reported that ambroxol can act as a “molecular chaperone” for the lysosomal enzyme beta-glucocerebrosidase (GC-ase, UniProtKB entry P04062), thereby increasing the quantity and activity of this enzyme. This may mean that ambroxol may be suitable for the treatment of Gaucher disease (Maegawa GHB et al., J Biol Chem 284(35):23502-23516, 2009), which is the most prevalent lysosomal storage disorder and is caused by GC-ase deficiency. Similarly, this ability to increase GC-ase activity may be beneficial in treating Parkinson's disease (PD) in individuals with loss-of-function mutations in the glucocerebrosidase gene GBA1 (McNeill A et al., Brain 137(5):1481-1495, 2014). Recent reports have shown that daily ambroxol administration can increase GC-ase activity in the brain of healthy non-human primates (Migdalska-Richards A et al., Synapse 71(7):e21967, 2017). Ambroxol has also been shown to increase the transcription factor EB (TFEB), a master regulator of autophagosome / lysosomal genes, leading to increases in other lysosomal proteins, including the protease cathepsin D. For example, see Magalhaes, J., Gegg, ME, Migdalska-Richards, A., Schapira, AH, 2018. Effects of ambroxol on the autophagy-lysosome pathway and mitochondria in primary cortical neurons. Sci Rep-uk 8, 1385. Therefore, ambroxol continues to be the subject of considerable research interest and effort. In the process of efforts directed towards identifying and developing new treatment methods and compositions based on ambroxol, the inventors designed novel modified forms of ambroxol (i.e., ambroxol analogs). These modified compounds are considered to be able to confer one or more advantages over one or more of ambroxol, ambroxol hydrochloride, and bromhexine, for example, better stability leading to an increase in half-life and duration of action in the body.

Prior Art Documents

Non-Patent Documents

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[0006] Summary of the Invention This summary is provided to introduce a selection of concepts in a simplified form that are further described below in the detailed description. This summary is not intended to identify key or essential features of the claimed subject matter, nor is it intended to be used to limit the scope of the claimed subject matter. Other features, details, utilities, and advantages of the claimed subject matter will become apparent from the following detailed description, which includes the illustrative aspects shown in the accompanying drawings and defined in the appended claims.

[0007] The present invention relates to analog forms of ambroxol and related compounds, compositions containing them, and methods for preventing and / or treating various diseases and medical conditions involving the administration of analogs of ambroxol and related compounds.

[0008] More specifically, the present invention relates to a compound according to formula I: [Chemical Formula] [wherein, ····· indicates that the associated R or X group may be attached to any available carbon atom on the phenyl ring, R a is H, hydroxyl (OH), lower alkyl (e.g., C such as CH3 and CH2CH3), 1~3(alkyl), and a lower alcohol (e.g., C such as CH2OH, etc.) 1~3 selected from R b is selected from H and lower alkyl (e.g., C such as CH3 and CH2CH3, etc.) 1~3 alkyl R c and R d are each independently selected from H, lower alkyl (e.g., C such as CH3 and CH2CH3, etc.) 1~3 alkyl), or R c and R d are each, respectively, a part of a 4-, 5-, 6- or 7-membered ring structure connecting R c and R d (e.g., -R c -N-R d -(CH2) n -, where n is an integer selected from 1, 2, 3 and 4), each of R 1 to R 14 is independently selected from H and D, X 1 and X 2 are independently selected from F, Cl, Br, I, lower alkyl (e.g., C such as CH3 and CH2CH3, etc.) 1~3 alkyl), lower alkoxy (e.g., C such as OCH3 and OCH2CH3, etc.) 1~3 alkoxy), lower alkylamine (e.g., C such as NR b CH3 and NR b CH2CH3, etc.) 1~3 alkylamine), lower acyl (e.g., C such as C(O)CH3 and C(O)CH2CH3, etc.) 2~4 acyl), nitro (NO2), nitrile (CN), sulfoxide (SO-R), sulfonate (SO2-R), and sulfate (O-SO2-O-R), provided that X 1 and X 2 are not both Br] or provides a pharmaceutically acceptable salt, solvate or prodrug thereof.

[0009] In a particular embodiment, the present invention relates to a compound according to formula Ia: [ka] [In the formula, R a , R b , R c , R d , R 1 From R 14 , X 1 and X 2 This is as described for the compound of formula I, however, X 1 and X 2 [It is not possible for both to be Br] Alternatively, the present invention provides a pharmaceutically acceptable salt, solvate, or prodrug thereof.

[0010] Where necessary, one or all of the H atoms in the compound of formula I may be replaced by deuterium atoms.

[0011] The analogs described herein unexpectedly demonstrate improved metabolic stability, which leads to an overall improvement in the cost of the analogs. In addition, the increased stability of the analogs may allow for higher exposure to the analogs when administered. Furthermore, the improved stability of the analogs may allow for reduced dosing schedules compared to ambroxol alone. An additional potential benefit of compounds following Formula I is resistance to metabolism (compared to ambroxol), for example, resistance to metabolism via the cleavage of carbon-nitrogen covalent bonds of linking groups between ring structures of compounds following Formula I (e.g., via oxidation).

[0012] The present invention also provides pharmaceutical compositions comprising a compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof), as well as methods for preventing and / or treating various diseases and medical conditions in a subject, involving the administration of a compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof).

[0013] More specifically, the present invention provides methods for preventing and / or treating diseases or medical conditions, selected from the group consisting of respiratory diseases and conditions (e.g., bronchopulmonary diseases, particularly those related to the production of excessive and / or highly viscous mucus), and including pain associated with acute sore throat, lysosomal storage disorders (LSD), e.g., Gaucher disease, neurological diseases and conditions (e.g., PD, and other age-related diseases with autophagy dysfunction) (Dockrill P., ScienceAlert, February, 2020).

[0014] In addition, the present invention provides methods for treating, reducing, and / or stabilizing symptoms associated with neurological diseases and conditions such as Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease, frontotemporal dementia, Pick's disease, Gaucher disease, and amyotrophic lateral sclerosis (i.e., Lou Gehrig's disease).

[0015] Furthermore, the present invention provides a method for extending the average life expectancy of a subject. Specifically, the compound of formula I may be used in a method for (a) treating, inhibiting or reducing aging in a subject, (b) treating, inhibiting or reducing age-related symptoms or age-related diseases in a subject, and / or (c) increasing the healthy lifespan, life expectancy and / or intellectual sharpness of a subject.

[0016] In a preferred embodiment, the subject is a mammal, and in a further preferred embodiment, the mammal is a human, a domesticated animal (e.g., a dog, a cat, a horse), or a livestock (e.g., a cow or a pig).

[0017] The object and features of the present invention can be better understood by referring to the following detailed description and accompanying drawings. [Brief explanation of the drawing]

[0018] [Figure 1] Figure 1 shows the structures of representative compounds (compounds 1 to 6) of the present invention.

[0019] [Figure 2] Figures 2A and 2B show the mean (± standard deviation) plasma and brain concentrations (ng / mL and ng / g, respectively) versus time (h) profiles in female C57BL / 6 mice orally administered a single dose of ambroxol hydrochloride (Figure 2A) versus ZW-010 HCl (Compound 1; also referred to herein as "ZW-010": 10 mg / kg, free base equivalent) (Figure 2B). These results demonstrate that halogenated analogs of ambroxol substantially improve exposure in brain tissue, making these compounds ideal for the treatment of the neurological disorders described herein.

[0020] [Figure 3] Figure 3 shows that both difluorosubstituted ambroxol (compound 1; also referred to herein as "ZW-010") and dimethoxysubstituted ambroxol (compound 5; also referred to herein as "ZW-011") increase lysosome size. To acquire data, human induced pluripotent stem cell (iPSC)-derived neurons were grown in culture medium for 14 days. Lysosomes were stained with "Lysotracker," cells were fixed, and imaged using a confocal microscope. Lysosome size was measured using IMARIS software (Bitplane).

[0021] [Figure 4] Figure 4 shows that both difluoro-substituted ambroxol (compound 1; also referred to herein as "ZW-010") and dimethoxy-substituted ambroxol (compound 5; also referred to herein as "ZW-011") promote lysosome and autophagosome as well as TFEB gene expression. To obtain data, N2A neuroblastoma cells were treated with the solvent alone (DMSO) or with 10 micromolar concentrations of ZW-010 (difluoro) or ZW-011 (dimethoxy) for 3 days. Cells were extracted and mRNA was quantified by qPCR.

[0022] [Figure 5] Figure 5 shows that both difluoro-substituted ambroxol (compound 1; also referred to herein as "ZW-010") and dimethoxy-substituted ambroxol (compound 5; also referred to herein as "ZW-011") promote lysosome and autophagosome gene expression to a greater extent in iPSC-derived human neurons compared to ambroxol. To obtain data, iPSC-derived human neurons were treated with solvent alone (DMSO), or with 10 micromolar concentrations of ZW-010 (difluoro) or ZW-011 (dimethoxy) for 3 days. Cells were extracted and mRNA was quantified by qPCR.

[0023] [Figure 6] Figures 6A-B show that difluoro-substituted ambroxol (compound 1; also referred to herein as "ZW-010") induces TFEB translocation to the nucleus (Figure 6A). To acquire data, human iPSC-derived neurons were treated with difluoro-substituted ambroxol for 14 days. Cells were fixed, immunostained for endogenous TFEB, and the nuclei were identified by DAPI staining. Cells were imaged using a Leica SP8 confocal microscope. TFEB signals appeared as dots throughout the cytoplasm and nucleus (Figure 6B). Nuclear TFEB dots were quantified using IMARIS software (Bitplane).

[0024] [Figure 7]Figures 7A and 7B show that difluoro-substituted ambroxol (compound 1; also referred to herein as "ZW-010") and dimethoxy-substituted ambroxol (compound 5; also referred to herein as "ZW-011") reduced the secretion of beta-amyloid species A beta-40 (Figure 7A) and A beta-42 (Figure 7B) into the culture medium of iPSC-derived human neurons associated with the Swedish amyloid precursor protein mutation. To obtain data, iPSC-derived human neurons associated with the Swedish mutation were treated with the solvent alone (DMSO), or with 10 micromolar concentrations of ambroxol ZW-010 (difluoro) or ambroxol ZW-011 (dimethoxy) for 5 days. The culture medium was collected and analyzed by ELISA assay.

[0025] [Figure 8] Figure 8 provides animal data demonstrating that ambroxol was able to increase lifespan in the mouse B6D2F1 model. Group 1 represents control animals that did not receive ambroxol, and Group 2 represents animals that received 50 mg / kg (body weight) of ambroxol daily as a chow supplement, starting at 2 months of age.

[0026] [Figure 9] Figure 9 shows results obtained from mice treated with ambroxol at 2 months and tested for novel location recognition at 7 months. This cognitive test measures short-term working memory involving the recognition of familiar objects found in unfamiliar locations (see Magen et al., Eur J Neurosci 35:870-882, 2012; Magen and Chesselet, J Parkinson's Disease 1:217-227, 2011). The discriminant index (DI) = (tnovel - tfamiliar) / (tnovel + tfamiliar) was used to evaluate the time spent exploring the vicinity of objects in a novel location ("tnovel") versus the total exploration time. A discriminant index score greater than zero is considered to indicate good location recognition memory.

[0027] [Figure 10] Figures 10A–D show the effect of ambroxol on basal macroautophagy in mouse cells. Mouse fibroblasts (NIH3T3 cells) expressing the tandem reporter mCherry-GFP-LC3 were exposed to the indicated concentration of ambroxol in complete medium for 24 hours. Figure 10A, schematic diagram of the autophagy compartment analyzed. Figures 10B–D, number of autophagy vacuoles (AV) (Figure 10B); autophagosomes (APG) (Figure 10C); and autolysosomes (AUT) (Figure 10D). All values ​​are mean + sem, and quantification was performed using high-content microscopy in three different experiments with at least 2,500 cells per condition. Differences from untreated (0 μM ambroxol) are significant at *p<0.05, **p<0.01, and ***p<0.001.

[0028] [Figure 11] Figures 11A-B show the results of ambroxol treatment in an AD mouse model. [Modes for carrying out the invention]

[0029] Detailed explanation I. Definition The following definitions are provided for the specific terms used in the descriptions below.

[0030] When used in specifications and claims, the singular forms “a,” “an,” and “the” include multiple references unless the context clearly indicates otherwise.

[0031] The present invention may "comprise" (open-ended) or "essentially consist of" the components of the present invention and other raw materials or elements described herein. As used herein, "comprising" means the enumerated elements, or their equivalents in structure or function, plus any other elements (singular or plural) not enumerated. The terms "having" and "including" shall also be interpreted as open-ended unless the context suggests otherwise. As used herein, "essentially consisting of" means that the present invention may include raw materials in addition to those enumerated in the claims, provided that the additional raw materials do not materially alter the basic and novel features of the claimed invention.

[0032] As used herein, “subject” means a vertebrate, preferably a mammal, more preferably a human, and even more preferably a domesticated animal, such as a pet or livestock. Mammals include, but are not limited to, rodents, primates, humans, livestock, sports animals, and pets. In other preferred embodiments, “subject” is a rodent (e.g., guinea pig, hamster, rat, mouse), a rodent (e.g., mouse), a canid (e.g., dog), a feline (e.g., cat), an equid (e.g., horse), a primate, a primate (e.g., monkey or ape), a monkey (e.g., marmoset, baboon), or an ape (e.g., gorilla, chimpanzee, orangutan, gibbon). In other embodiments, non-human mammals, particularly mammals conventionally used as models to demonstrate therapeutic efficacy in humans (e.g., rodents, primates, pigs, canids, or rabbits), may be used. In a preferred embodiment, “individual” or “patient” (as in the case of a subject of treatment) means a mammal, in particular a non-human primate, such as apes and monkeys, and most specifically a human.

[0033] As understood herein, “effective amount” of the pharmaceutical composition of the present invention means an amount of the composition suitable for inducing a therapeutically beneficial response in a subject, for example, promoting mucus clearance associated with respiratory diseases and conditions, relieving pain associated with acute sore throat, relieving symptoms associated with lysosomal storage disorders (LSD) and neurological diseases and conditions, or extending and / or increasing and / or improving healthy lifespan, lifespan and / or intellectual sharpness, for example, increasing survival and / or healthy aging and / or reducing morbidity or age-related diseases in a subject.

[0034] The term “dose” or “dosage” as used herein refers to a physically discontinuous unit suitable for administration to a subject, each dosage containing a predetermined amount of the active pharmaceutical ingredient calculated to produce a desired response.

[0035] The terms “about” or “approximately” mean an acceptable range for a particular value as determined by those skilled in the art, which is partly determined by how the value is measured or determined, for example, by the limitations of the measuring system. For example, “about” can mean a range of up to 20%, preferably up to 10%, more preferably up to 5%, and even more preferably up to 1% of a given value. Alternatively, particularly with respect to biological systems or biological processes, the term can mean within one order of magnitude of the value, preferably up to five times, and more preferably up to two times. Unless otherwise stated, the term “about” means an acceptable range of error for a particular value, for example, within ±1 to 20%, preferably ±1 to 10%, and more preferably ±1 to 5%. In a further embodiment, “about” should be understood to mean + / - 5%.

[0036] Where a range of values ​​is provided, it is understood that each intermediate value between that range and any other described range's upper and lower limits, or intermediate values ​​within that described range, are included within the present invention. These smaller ranges' upper and lower limits may independently be included within smaller ranges, depending on any specifically excluded limit values ​​within the described range, and are also included within the present invention. Where a described range includes one or both of the limit values, ranges excluding either or both of those included limit values ​​are also included within the present invention.

[0037] All percentages and ratios used herein are by weight of the total composition unless otherwise indicated herein. All temperatures are in degrees Celsius unless otherwise specified. All measurements performed are at 25°C and atmospheric pressure unless otherwise specified.

[0038] All ranges listed herein (including those listing ranges "between" two values) include endpoints. For example, terms such as "about," "generally," "substantially," and "approximately" modify the term or value and, as a result, should be interpreted as not absolute but not inferred from the prior art. Such terms will be defined by the circumstances and the terms they modify, for these terms will be understood by those skilled in the art. This includes, at a minimum, the degree of expected experimental error, technical error, and instrument error with respect to a given technique used to measure the value.

[0039] As used herein, the term "and / or" in a list of two or more items means that any one of the listed features may be present, or any combination of two or more of the listed features may be present. For example, if a composition of the present invention is described as containing features A, B, and / or C, the composition may contain feature A alone; feature B alone; feature C alone; a combination of A and B; a combination of A and C; a combination of B and C; or a combination of A, B, and C.

[0040] As used herein, the term “lower alkyl” includes linear alkyl groups, branched alkyl groups, and cyclic alkyl groups having 1 to 8 carbon atoms (e.g., methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, hexyl, cyclopropyl, cyclobutyl, cyclopentyl, etc.).

[0041] As used herein, the term “lower alcohol” includes alcohol groups comprising linear, branched, or cyclic alkyl groups having 1 to 8 carbon atoms and one or more hydroxyl (OH) groups (e.g., methanol, ethanol, propanol, etc.).

[0042] The term “pharmaceutically acceptable salt,” as used herein, refers to a salt of a compound of formula I that retains the desired biological activity, and includes pharmaceutically acceptable acid addition salts and base addition salts. Suitable pharmaceutically acceptable acid addition salts of compounds of formula I can be prepared from inorganic or organic acids. Examples of such inorganic acids are hydrochloric acid, sulfuric acid, and phosphoric acid. Suitable organic acids may be selected from the aliphatic, alicyclic, aromatic, heterocyclic, carboxylic acid, and sulfonic acid classes, examples of which are formic acid, acetic acid, propionic acid, succinic acid, glycolic acid, gluconic acid, lactic acid, malic acid, tartaric acid, citric acid, fumaric acid, maleic acid, alkylsulfonic acid, and arylsulfonic acid. Additional information on pharmaceutically acceptable salts can be found in Remington's Pharmaceutical Sciences, 19th Edition, Mack Publishing Co, Easton PA 1995.

[0043] The term "solvate" refers to any form of the compound of formula I that results from solvation with a suitable solvent. Such a form may be, for example, a crystalline solvate or a complex that can be formed between the solvent and the dissolved compound.

[0044] The term "prodrug" usually refers to a compound that is converted to a compound of formula I in a biological system by metabolic means (e.g., hydrolysis, reduction, or oxidation). For example, an ester prodrug of a compound of formula I containing a hydroxyl group may be convertible to a compound of formula I in vivo by hydrolysis. Suitable esters of compounds of formula I containing a hydroxyl group may be, for example, acetate esters, citrate esters, lactate esters, tartaric acid esters, malonic acid esters, oxalic acid esters, salicylic acid esters, propionic acid esters, succinic acid esters, fumarate esters, maleic acid esters, methylene-bis-p-hydroxynaphthoate esters, gestisates, isethionate esters, di-p-toluyl tartaric acid esters, methanesulfonic acid esters, ethanesulfonic acid esters, benzenesulfonic acid esters, p-toluenesulfonic acid esters, cyclohexylsulfamic acid esters, and quinic acid esters. As another example, ester prodrugs of compounds of formula I containing a carboxyl group may be convertible to compounds of formula I in vivo by hydrolysis. Examples of ester prodrugs include those described by Leinweber FJ, Drug Metab Rev 18:379-439 (1987). Similarly, acyl prodrugs of compounds of formula I containing an amino group may be convertible to compounds of formula I in vivo by hydrolysis. Examples of prodrugs for these and other functional groups (including amines) are provided in Prodrugs: challenges and rewards, Valentino J Stella (ed), Springer, 2007.

[0045] When the term "derivative" is used herein, it refers to a second compound derived from a first compound (for example, a brominated version of a non-brominated parent compound). For example, ambroxol is a derivative of its parent compound, bromhexine.

[0046] As used herein, “life expectancy” means the time until death. As used herein, “healthy life expectancy” or “healthy aging” means the time of life free from (or optimally free from) serious illness. As used herein, “intellectual acuity” is a measure of a subject’s cognitive abilities, such as concentration, attention span, and clarity.

[0047] As used herein, “nutrient sensing” is the ability of a cell to sense and respond to changes in nutrient levels, as described in Efeyan et al., “Nutrient Sensing Mechanisms and Pathways,” Nature 517: 302-310 (2015) (which is incorporated herein by reference in its entirety).

[0048] As used herein, “treating, inhibiting, and / or reducing aging, age-related symptoms, and / or age-related diseases” means reducing the risk of developing, delaying the onset of, slowing the progression of, and / or reducing the severity and / or manifestation of signs and / or degenerative disorders of aging, and includes, but is not limited to, preventing the onset, development or progression of signs and / or degenerative disorders of aging.

[0049] The term “pharmaceutically acceptable carrier” as used herein means any carrier, diluent or excipient that is compatible with the other components of the composition and is not harmful to the subject intended to receive the composition. Compound of the present invention

[0050] The compounds of the present invention are analog forms of ambroxol and related compounds (including bromhexine and ambroxol hydrochloride), as defined by Formula I. Table 1 shows the structures of ambroxol (also known by its chemical name trans-4-((2-amino-3,5-dibromobenzyl)amino)cyclohexanol), bromhexine (also known by its chemical name 2-amino-3,5-dibromo-N-cyclohexyl-N-methylbenzenemethaneamine), and ambroxol hydrochloride.

[0051] [Table 1]

[0052] In some embodiments, the compound is selected from halogen or alkoxy analogs of ambroxol and bromhexine shown in Figure 1. These compounds are thought to exhibit greater resistance to metabolism (compared to the corresponding ambroxol or bromhexine compounds), such as metabolism via cleavage of carbon-nitrogen covalent bonds in linking groups between ring structures of compounds according to formula I (e.g., by oxidation), or oxidation of carbon-hydrogen bonds in aromatic rings to form phenolic metabolites. In other words, these compounds may have superior stability.

[0053] The compounds of the present invention may be used in methods for preventing and / or treating various diseases and medical conditions, including respiratory diseases and conditions, lysosomal storage disorders (LSDs), and neurological diseases and conditions, and may be used in methods for extending the life expectancy of subjects. In such methods, the compounds of the present invention, which exhibit superior stability (compared to, for example, corresponding compounds lacking deuteration), may exhibit, for example, one or more advantageous pharmacokinetic properties. Preparation of the compound of the present invention

[0054] The compounds of the present invention may be prepared by methods known to those skilled in the art of organic synthesis. For example, U.S. Patent Application Publication US2004 / 0242700, which is incorporated herein in its entirety by reference, provides a synthetic protocol for the preparation of ambroxol. This protocol can be readily adapted to enable the synthesis of analog forms of ambroxol and related compounds. In addition, a protocol for the synthesis of ambroxol and related compounds, including deuterium-containing analogs, is disclosed in Latli B et al., J Label Compd Radiopharm 53:15-23, 2010 (which is incorporated herein in its entirety by reference), which can also be readily adapted for the preparation of compounds of formula I. For example, "Scheme 4" of Latli et al., 2010 can be adapted for the preparation of its compound by replacing 2-amino-3,5-dibromobenzaldehyde with the corresponding analog of the compound of formula I. Furthermore, the compounds of the present invention may be prepared by methods such as those exemplified herein. Salt of the compound of the present invention

[0055] For compounds typically containing acidic or basic groups (such as carboxyl or amino groups), such groups are not necessarily in the form of free acid or free base. When referring to the compounds of the present invention, the reference is intended to include the salt forms of the compounds. Therefore, within the scope of the present invention are salts of the compounds of formula I. Preferred salts are pharmaceutically acceptable salts.

[0056] The term "salt" includes salts added to free acids or free bases. The term "pharmaceutically acceptable salt" refers to a salt that possesses a toxicity profile within a range that yields usefulness in pharmaceutical applications. Pharmaceutically unacceptable salts may still possess properties such as high crystallinity, which may have usefulness in the practice of the present invention, for example, in the processes of synthesis, purification, or formulation of therapeutic compounds.

[0057] Suitable pharmaceutically acceptable acid addition salts can be prepared from inorganic or organic acids. Examples of inorganic acids include hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, carbonic acid, sulfuric acid, and phosphoric acid. Suitable organic acids may be selected from aliphatic, alicyclic, aromatic, araliphatic, heterocyclic, carboxylic acid, and sulfonic acid classes, examples of which include formic acid, acetic acid, propionic acid, succinic acid, glycolic acid, gluconic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, glucuronic acid, maleic acid, fumaric acid, pyruvic acid, aspartic acid, glutamic acid, benzoic acid, anthranilic acid, and 4-hydro This includes xybenzoic acid, phenylacetic acid, mandelic acid, embonic acid (pamoic acid), methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, pantothenic acid, trifluoromethanesulfonic acid, 2-hydroxyethanesulfonic acid, p-toluenesulfonic acid, sulfanilic acid, cyclohexylaminosulfonic acid, stearic acid, alginic acid, β-hydroxybutyric acid, salicylic acid, galactaric acid, oxalic acid, malonic acid, and galacturonic acid. Examples of pharmaceutically unacceptable acid addition salts include, for example, perchlorates and tetrafluoroborates. All of these acid addition salts can be prepared from compounds of formula I, for example, by reacting a suitable acid with a specific compound.

[0058] Suitable pharmaceutically acceptable base addition salts of the compounds of formula I include, for example, metal salts, including alkali metal salts, alkaline earth metal salts, and transition metal salts, such as calcium salts, magnesium salts, potassium salts, sodium salts, and zinc salts. Also included are organic salts made from basic amines, such as, for example, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine), and procaine. Examples of pharmaceutically unacceptable base addition salts include lithium salts and cyanates. All of these base addition salts can be prepared from the compounds of formula I, for example, by reacting a suitable base with a specific compound. Method using the compound of the present invention

[0059] As mentioned above, the compounds of the present invention may be used in methods for preventing and / or treating various diseases and medical conditions, including respiratory diseases and conditions, lysosomal storage disorders (LSD), and neurological diseases and conditions, and may also be used in methods for extending the life expectancy of a subject. For example, the compounds of formula I may be used in methods for (a) treating, inhibiting or reducing aging in a subject, (b) treating, inhibiting or reducing age-related symptoms or age-related diseases in a subject, and / or (c) increasing the healthy lifespan, life expectancy and / or intellectual sharpness of a subject.

[0060] In some other embodiments, the present invention more specifically relates to a method for preventing and / or treating a disease or medical condition in a subject, wherein the disease or condition is selected from the group consisting of respiratory diseases and conditions (e.g., bronchopulmonary diseases, and in particular those related to the production of excessive and / or highly viscous mucus), and includes pain associated with acute sore throat, lysosomal storage disorders (LSD), e.g., Gaucher disease, and neurological diseases and conditions (e.g., Parkinson's disease (PD), Alzheimer's disease (AD), Huntington's disease, frontotemporal dementia, Pick's disease, Gaucher disease, and amyotrophic lateral sclerosis (i.e., Lou Gehrig's disease) and other age-related diseases with autophagy dysfunction), wherein the method comprises the step of administering to the subject an effective amount of a compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof). The present invention also relates to the use of compounds of formula I (or pharmaceutically acceptable salts, solvates, or prodrugs thereof) as pharmaceuticals for the prevention and / or treatment of diseases or medical conditions selected from respiratory diseases and conditions, lysosomal storage disorders (LSDs), and neurological diseases and conditions. In addition, the present invention relates to pharmaceutical compositions for the treatment of diseases or medical conditions selected from respiratory diseases and conditions, lysosomal storage disorders (LSDs), and neurological diseases and conditions, comprising a therapeutically effective amount of a compound of formula I (or pharmaceutically acceptable salts, solvates, or prodrugs thereof) together with a pharmaceutically acceptable carrier.

[0061] In some other embodiments, the present invention relates to a method for sustaining the healthy lifespan, lifespan, and / or intellectual sharpness of a subject, comprising the step of administering to the subject an effective amount of the compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof). The present invention also relates to the use of the compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) as a pharmacopoeia for extending life expectancy and / or reducing diseases or symptoms associated with aging or age. In addition, the present invention relates to a pharmacopoeia composition for a life expectancy extension treatment, comprising a therapeutically effective amount of the compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) together with a pharmaceutically acceptable carrier.

[0062] In some further embodiments, the present invention relates to a long-term method for increasing and / or improving the healthy lifespan, lifespan and / or intellectual sharpness of a subject, the method comprising the step of administering a therapeutically effective amount of a compound of formula I (or a pharmaceutically acceptable salt, solvate or prodrug thereof), wherein the administration of the compound is over a period of at least 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 years. Using such a method, the lifespan, healthy lifespan, intellectual sharpness and / or healthy aging of the subject is preferably extended, improved or accelerated by up to about 10%, 20%, 30%, 40%, 50%, 60%, or 70% compared to an untreated control subject.

[0063] Examples of age-related diseases and conditions include, but are not limited to, cardiovascular diseases, metabolic syndrome, osteoporosis, neurodegenerative diseases, prediabetes, diabetes, obesity, osteoporosis, coronary artery disease, cerebrovascular disease, heart attack, stroke, peripheral artery disease, aortic valve disease, stroke, mild cognitive impairment, predementia, dementia, macular degeneration and cataracts, hair loss, graying hair, loss of motor function, decreased stamina, fatigue, increased susceptibility to infection, metabolic changes, biochemical changes, cardiac hypertrophy, heart failure, myocardial infarction, ischemia-reperfusion injury, inflammatory diseases, pro-inflammatory conditions, arthropathy, autoimmune diseases, and / or Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease, frontotemporal dementia, Pick's disease, Gaucher disease and amyotrophic lateral sclerosis (i.e., Lou Gehrig's disease).

[0064] In the method of the present invention, the above compound may be administered to the subject in a daily dose of approximately 20-500 mg / day, 50-150 mg / day, 50-200 mg / day, 50-250 mg / day, 250-500 mg / day, or 250 mg-1500 mg / day. Higher or lower doses are also intended, as it may be necessary to use doses outside these ranges in some cases. The daily dose may be divided, for example, into two to four equal daily doses. Furthermore, since long-term administration may be required in a manner that induces, increases, and / or improves the subject's healthy lifespan, life expectancy, and / or intellectual sharpness, the above compounds are preferably administered in doses of approximately 50 mg / day, 75 mg / day, 100 mg / day, 150 mg / day, 200 mg / day, 250 mg / day, 300 mg / day, 350 mg / day, 400 mg / day, 450 mg / day, 500 mg / day, 550 mg / day, 600 mg / day, 650 mg / day, 700 mg / day, 750 mg / day, 800 mg / day, 850 mg / day, 900 mg / day, 950 mg / day, 1000 mg / day, 1050 mg / day, 1100 mg / day, 1150 mg / day, 1200 mg / day, and 12 It may be administered in doses of 50 mg / day, 1300 mg / day, 1350 mg / day, or between 50 and 150 mg / day, 50 and 200 mg / day, 50 and 250 mg / day, 250 and 500 mg / day, 250 and 1000 mg / day, 1000 and 1500 mg / day, 1500 and 2000 mg / day, or between 1000 and 2000 mg / day, or less than 1000 mg / day, or approximately 1 mg / kg / day, 2 mg / kg / day, 3 mg / kg / day, 4 mg / kg / day, 5 mg / kg / day, 6 mg / kg / day, 7 mg / kg / day, 8 mg / kg / day, 9 mg / kg / day, 10 mg / kg / day, 11 mg / kg / day, 12 mg / kg / day, and / or between 4 and 12 mg / kg / day.

[0065] In some preferred embodiments, the methods of the present invention may be particularly applied to subjects including humans suffering from pain associated with acute sore throat, humans suffering from bronchopulmonary diseases associated with the production of excessive and / or highly viscous mucus, lysosomal storage disorders (LSD), e.g., Gaucher disease, and humans suffering from neurological disorders and conditions, e.g., parkinsonism (including PD and Lewy body dementia), AD, ALS, HD, Pick's disease, Gaucher disease, and / or frontotemporal dementia (FTD). Parkinson's disease and other diseases with autophagy dysfunction

[0066] Ambroxol (and ambroxol hydrochloride and bromhexine) and the ambroxol analogs of the present invention may be particularly applied to the treatment of age-related diseases with proteolysis / lysosome, autophagy dysfunction and chronic inflammation, such as Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease, frontotemporal dementia, Pick's disease, Gaucher disease, and amyotrophic lateral sclerosis (ALS) (i.e., Lou Gehrig's disease).

[0067] As mentioned above, ambroxol's ability to increase GC-ase activity may be beneficial in treating PD in individuals with loss-of-function mutations in the glucocerebrosidase gene GBA1 (McNeill A et al., 2014, above), which is an important heritable risk factor for PD (Do J et al., Neurodegener 14:36, 2019). The enhancement of GC-ase activity induced by ambroxol treatment is predicted to increase lysosomal degradation capacity and support the clearance of intracellular aggregated α-synuclein (aSyn), a protein involved in the pathogenesis of PD (Migdalska-Richards A et al., 2016, above). GC-ase deficiency is also associated with lysosomal and mitochondrial dysfunction observed in PD (Brooker and Krainc, 2021 Essays in Biochemistry, 65(7), pp. 873-883), which may partially explain the improvement of lysosomal and mitochondrial function by ambroxol (Magalhaes et al., 2018 Scientific Reports, 8(1), p. 1385). Based on this data and promising preclinical data, the use of ambroxol is being investigated in clinical trials as a disease-modifying compound for PD (see, e.g., Silveira CRA et al., BMC Neurol 19:20, 2019, and Mullin S et al., JAMA Neurol 77:427-434, 2020) and Gaucher disease, a lipid storage disorder (Zimran, Altarescu and Elstein, 2013 Blood Cells, Molecules & Diseases, 50(2), pp. 134-137; Narita et al., 2016 Annals of Clinical and Translational Neurology, 3(3), pp. 200-215).

[0068] However, it is well known that the potentially beneficial bioactivity of ambroxol extends far beyond its chaperoning of GC-ases. In particular, ambroxol is involved in the autophagic endolysosomal network (AELN) (McNeill et al., 2014 Brain: A Journal of Neurology, 137(Pt 5), pp. 1481-1495; Fois et al., 2015 Cell Calcium, 58(6), pp. 628-637; Magalhaes et al., 2018 above), the induced immune response (Beeh et al., 2008 European Journal of Medical Research, 13(12), pp. 557-562; Kern and Schwickert, 2017 Journal of Pain Research, 10, pp. 1905-1929), and the blocking of channels involved in chronic pain (Russo et al., 2022 Pain [Preprint]). Ambroxol exerts broad-spectrum effects (available at doi.org / 10.1097 / j.pain.0000000000002693). Therefore, as a core feature, ambroxol has been potentially demonstrated to be useful in treating a wide variety of diseases with defects in AELN, the immune system, or nociception. For example, many neurodegenerative disorders, including PD, FTD, ALS, HD, and AD, exhibit deficiencies in the production or degradation of neurotoxic protein species within the AELN, which in turn can lead to a chronic inflammatory environment in brain tissue.

[0069] Some of the mechanisms by which ambroxol exerts its influence on cellular processes have been elucidated. Ambroxol is an amphiphilic amine that readily traverses the cell membrane via passive diffusion, thereby gaining direct access to intracellular and organelle spaces. Intracellularly, ambroxol acts as a weak base, protonated and captured in acidic intracellular compartments, including lysosomes and other structures of the late endolysosomal pathway (Magalhaes J et al., 2018 above; and Fois G et al., 2015 above). These properties explain ambroxol's high bioavailability and its tendency to accumulate in lipid-rich organ systems such as the brain, lungs, and skin (Mullin S et al., 2020 above). In the cytoplasm, ambroxol also acts as a potent scavenger and stabilizer of free radicals generated during cellular metabolism and as part of the innate immune response (Stetinova, Herout and Kvetina, 2004 Clinical and Experimental Medicine, 4(3), pp. 152-158). Finally, ambroxol has been shown to confer analgesia by directly blocking Nav1.7 / 8 channels associated with neuropathic pain (Kern and Schwickert, 2017, above).

[0070] The amphipathic capture of ambroxol in the acidic compartment exerts several effects on the AENL system. One important consequence of ambroxol protonation in lysosomes is the resulting deoxidation of the lysosomal lumen (Mullin S et al., 2020, and Lu S et al., PloS One 12:e0173771, 2017). This process mobilizes calcium reservoirs within intracellular organelles, which leads to the activation of lysosome-associated transcription factor EB (TFEB). Activated TFEB relocalizes to the nucleus, where it reinitiation of the gene network transcription using its master regulatory control over lysosomal biosynthesis (Medina DL et al., Nat Cell Biol 17:288-299, 2015). Upregulation of this gene network leads to increased cellular activity and degradation of long-lived proteins (Sardiello et al., 2009 Science, 325(5939), pp. 473-477), particularly proteins associated with neurodegenerative disorders such as hyperphosphorylated tau (Martini-Stoica et al., 2018 The Journal of Experimental Medicine, 215(9), pp. 2355-2377). Ambroxol effectively activates TFEB in both cell culture and in vivo. In cultures derived from the brain (Magalhaes et al., 2018, see above), skin (McNeill et al., 2014, see above), or immune-related tissues (Choi et al., 2018 Antimicrobial Agents and Chemotherapy, 62(9)), application of ambroxol results in upregulation of TFEB at the transcriptional, protein, and / or activation levels. Increasing TFEB activity promotes the expression of multiple lysosomal enzymes, including the protease cathepsin D, which is known to degrade alpha-synuclein, beta-amyloid, and tau.For example, see Suire, CN et al. "Cathepsin D: A Candidate Link between Amyloid β-protein and Tauopathy in Alzheimer Disease" J Exp Neurology 2, 10-15 (2021); Sevlever, D. et al., "Cathepsin D Is the Main Lysosomal Enzyme Involved in the Degradation of α-Synuclein and Generation of Its Carboxy-Terminally Truncated Species," Biochemistry-US 47, 9678-9687 (2008).

[0071] In this regard, ambroxol in high-dose regimens can promote cellular autophagy to secretory regimens (see, e.g., McNeill A et al., 2014 above, and Silveira CRA et al., 2019 above). Secretory autophagy involves a mechanism in which normally functioning and defective proteins are encapsulated in membrane-bound organelles called autophagosomes and secreted extracellularly, representing an alternative process to degradative autophagy in which autophagosomes fuse with lysosomes to promote the disruption and regeneration of the autophagosome's contents (Rabouille C et al., J Cell Sci 125:5251-5255, 2012). Upregulation of autophagy-dependent secretion is recognized as a potential route for disease modification in neurodegenerative disorders (Ponpuak M et al., Curr Opin Cell Biol 35:106-116, 2015).

[0072] In cases of Parkinson's disease (PD) and Alzheimer's disease (AD), the aggregate-prone proteins aSyn (in PD; see, e.g., Dehay B et al., J Neurosci Off J Soc Neurosci 30:12535-12544, 2010) and beta-amyloid (in AD; see, e.g., Vickers JC et al., Exp Neurol 141:1-11, 1996) form degradation-resistant inclusions that accumulate within autophagy vacuoles. In both cases, protein aggregates promote abnormal regulation at multiple points along the cellular autophagy pathway, which can potentially be mitigated by secretory efflux.

[0073] Ambroxol can directly participate in the secretory autophagy system. In primary neuronal cell cultures derived from genetically engineered cells expressing PD pathology and unmodified counterparts, ambroxol enhances aSyn secretion while clearing intracellular storage of an aggregate-prone version of this protein (Magalhaes J et al., 2018, above). These results further extend to AD pathology, where clearance of intracellular aggregate-prone tau is promoted by the application of bromhexine to brain-derived cell cultures (Chauhan S et al., Nat Commun 6:8620, 2015). In summary, these data indicate that ambroxol exerts beneficial effects on AELN by increasing innate cellular decomposition while removing toxic protein aggregates from cells such as neurons that possess lower innate decomposition capabilities.

[0074] Another important system for understanding the effects of ambroxol on human health is the immune response. Epidemiological and preclinical studies indicate that driving the immune response away from innate inflammatory responses towards an anti-inflammatory / adaptive context may be beneficial in several chronic disease situations. For example, in many neurodegenerative disorders, longitudinal sampling of patient serum and cerebrospinal fluid points to a gradual increase in inflammatory response markers such as cytokines including interleukin 1β, 6, 8, and tissue necrosis factor alpha, as well as activation of NOD-, LRR-, and pyrin domain-containing protein 3 (NLRP3)-related inflammasomes, reflecting cognitive loss (Heneka et al., 2015 The Lancet. Neurology, 14(4), pp. 388-405; Wang, Liu and Zhou, 2015 Translational Neurodegeneration, 4, p. 19). Consistently, variants in many genes with innate immunomodulatory functions are associated with an increased risk of AD and PD (Hollingworth et al., 2011 Nature Genetics, 43(5), pp. 429-435; Griciuc and Tanzi, 2021 Current Opinion in Neurology, 34(2), pp. 228-236). In contrast, upmodulation of anti-inflammatory and adaptive immune markers such as interferon-gamma and interleukin-12 correlates with a reduced incidence of AD in older adults (Yang et al., 2022 Alzheimer's & Dementia, 18(4), pp. 645-653).

[0075] Neurodegenerative disorders symbolize a chronic inflammatory condition that arises when the immune system is unable to mitigate immune sensitizers over a chronic timeframe. In cases of neurodegenerative disorders, these immune damages may be partially or entirely provided by toxic aggregate proteins produced by cells in the brain. Chronic inflammation can also occur under conditions of persistent infection, such as mycoplasma pneumonia and inappropriate immune activation by autologous antigens.

[0076] Regarding inflammation, ambroxol has also been shown to significantly restore the immune response against both pathogenic and autologous threats. Studies spanning multiple organ systems, particularly the brain (Jiang et al., 2020 BioMed Research International, 2020, p. e8131286), lungs (Takeda et al., 2016 Immune Network, 16(3), pp. 165-175; Zhang et al., 2016; Kokai et al., 2021 Microorganisms, 9(4), p. 880), and gut (Schneider et al., 2021 EMBO Molecular Medicine, 13(1), p. e12724; Cavalu et al., 2022 The FASEB Journal, 36(9), p. e22496), have confirmed that ambroxol acts to reduce pro-inflammatory responses to such threats while preserving the adaptive aspects of the immune response. In particular, ambroxol has been shown to reduce the expression of pro-inflammatory cytokines such as interleukin 1β, 6, 8, 10, and tissue necrosis factor-α (Bianchi et al., 1990 Agents and Actions, 31(3-4), pp. 275-279; Jang et al., 2003 Pharmacology & Toxicology, 92(4), pp. 173-179; Wang et al., 2011; Zhongguo Ying Yong, et al., Chinese Journal of Applied Physiology, 27(2), pp. 231-235), as well as reduce the activity of nuclear factor kappa B (NF-κB) and other upstream pathways of inflammasome activation (Cavalu et al., 2022, see above).This suppression is at least partially due to ambroxol's ability to capture free radicals (Peroni et al., 2013 International Journal of Immunopathology and Pharmacology, 26(4), pp. 883-887), and may also be accompanied by direct blocking of specific inflammation-related channels (Schneider et al., 2021, see above).

[0077] Unlike nonspecific immunosuppressants such as nonsteroidal anti-inflammatory drugs (NSAIDs), ambroxol preserves, and in some cases enhances, the anti-inflammatory and adaptive immune system aspects. This includes the upmodulation of the anti-inflammatory cytokines interleukin 10 and 12, as well as adaptive immune-related interferon-γ, in lung tissue in response to pathogens and ovalbumin sensitization (Takeda et al., 2016 above; Kokai et al., 2021 above). These immune effectors can explain the long-known clinical benefits of ambroxol in chronic respiratory diseases such as chronic obstructive pulmonary injury (Plomer and de Zeeuw, 2017 MMW Fortschritte der Medizin, 159(Suppl 5), pp. 22-33). In addition, these may be major factors in the preclinical observation of reduced inflammation in a model of ulcerative colitis (Schneider et al., 2021, see above) and reduced microglial activation in a model of intracerebral hemorrhage (Jiang et al., 2020, see above).

[0078] The anti-inflammatory effects described above are likely related to the effects of ambroxol on autophagy. Autophagy has been observed to play a central role in regulating the immune response. In particular, activation of TFEB function promotes the degradation of key mediators of inflammation, including inflammasome components such as NLRP3 and apoptosis-related speck-like proteins (ASCs) containing caspase recruitment domains (Shi et al., Nature Immunology, 13(3), pp. 255-263 (2012); Deretic, Immunity, 54(3), pp. 437-453 (2021)). Inflammasome inactivation via degradation leads to attenuation of the inflammatory response, including a reduction in the production and release of pro-inflammatory cytokines such as IL-1β. Therefore, the immunomodulatory function of ambroxol is likely downstream of its effects on autophagy.

[0079] Ambroxol also possesses potent analgesic properties. Mechanistically, this effect is due to ambroxol's ability to block voltage-gated sodium channels, including Nav1.7 (Leffler, Reckzeh and Nau, 2010 European Journal of Pharmacology, 630(1-3), pp. 19-28) and 1.8 (Weiser and Wilson, 2002 Molecular Pharmacology, 62(3), pp. 433-438), which are preferentially expressed by neurons involved in nociception (Bennett et al., 2019 Physiological Reviews, 99(2), pp. 1079-1151). This mechanism at least partially explains the classic use of ambroxol as a treatment for moderate to severe pain associated with acute pharyngitis (Fischer et al., 2002 Arzneimittel-Forschung, 52(4), pp. 256-263). In addition, there is growing interest in the use of ambroxol for the treatment of neuropathic pain (Russo et al., 2022, see above).

[0080] Furthermore, a recently discovered method by which cells regulate TFEB nuclear access and transcriptional function is poly(ADP-ribosyl)ation (parsylation) (Kim et al., 2021; Chen et al., 2024). Parsylation is a reversible post-translational modification process catalyzed by poly(ADP-ribose) polymerase (PARP) in the transfer of ADP-ribose units to a target protein. This modification mobilizes E3 ligase ubiquitination of the target protein, ultimately resulting in increased metabolic turnover via the ubiquitin-proteasome system (Vivelo et al., 2019).

[0081] In the case of TFEB, parsylation promotes nuclear localization in a phosphate-independent manner. As expected, nuclear localization enhances transcription initiated by TFEB, but with a significant caveat. In parsylated conditions, TFEB complexes with Wnt / beta-catenin, resulting in transcription initiation of a completely different set of genes than those in the CLEAR network (Kim et al., 2021). Essentially, parsylation allows TFEB to flexibly alter the protein, enabling it to be activated based on the instantaneous needs of the cell.

[0082] Several unexplained observations in brain cells treated with ambroxol may be explainable if ambroxol is involved in TFEB-mediated Wnt / beta-catenin pathway regulation. For example, after stroke in a mouse model (dMCAO example), ambroxol preferentially differentiates cells generated from the brain's proliferative zone (called the subventricular zone) into new neurons rather than more typical reactive astrocytes. In a parallel in vitro model (oxygen-glucose depletion in iPSCs), the authors show that ambroxol increases beta-catenin signaling and GC-ase (Ge et al., 2021). This is consistent with the predicted effect that Wnt / beta-catenin transcriptional regulation tends to assign the neuronal fate rather than glial fate to neural progenitor cells (Gao et al., 2021; Kriska et al., 2021).

[0083] Other studies support a link between ambroxol and Wnt / beta-catenin mediated via TFEB. One study demonstrated a link between GC-ase and Wnt / beta-catenin transcriptional regulation. In iPSC strains induced to a dopaminergic fate, GD mutations drive a reduction in Wnt / beta-catenin signaling. Addition of recombinant GC-ase restored Wnt / beta-catenin signaling to wild-type levels (Awad et al., 2017).

[0084] Therefore, in conclusion, while we do not wish to be constrained by theory, the aforementioned observations suggest that analogs described herein, such as ambroxol, may increase the degradation and / or secretion of toxic intracellular proteins, protein fragments, misfolded proteins, protein aggregates, or debris associated with AELN impairment. These analogs may further modulate the immune response to relatively low-inflammation situations, possibly via free radical capture and interaction with inflammation-related channels. Finally, the described analogs may block upregulated channels in disease situations characterized by chronic pain.

[0085] Significance - The ambroxol analogs described herein offer considerable potential as effective treatments for neurological disorders and conditions such as PD, AD, and other age-related disorders with autophagy dysfunction by enabling the secretion of toxic aggregated proteins (e.g., aggregated aSyn, beta-amyloid, and tau proteins) from affected cells despite autophagy dysfunction, and by maintaining the health of organelles.

[0086] In some cases, it may be advantageous to administer the ambroxol analogs described herein in conjunction with an active agent intended, for example, to prevent and / or remove toxic aggregated proteins.

[0087] Therefore, in another aspect of the present invention, a method is provided for preventing, reducing the symptoms of, and / or stabilizing the progression of, Alzheimer's disease (AD) or other diseases associated with pathogenic protein misfolding, aggregation, and deposition (including Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS) (i.e., Lou Gehrig's disease), Pick's disease, Gaucher's disease, and frontotemporal degeneration (FTD)) in a subject, comprising the step of administering to the subject an effective amount of ambroxol (or related compounds such as ambroxol hydrochloride and bromhexine) or an analog of the present invention in combination with one or more suitable anti-beta-amyloid antibodies or fragments thereof. Such combination therapy may produce, for example, a synergistic effect in removing aggregated beta-amyloid associated with AD.

[0088] Ambroxol analogs and anti-beta-amyloid antibodies or fragments thereof may be administered as part of the same treatment protocol, for example, as separate compositions. Here, the ambroxol analogs and anti-beta-amyloid antibodies or fragments thereof may be administered simultaneously or sequentially in any order (for example, within a few seconds or minutes or even a few hours (e.g., 2 to 48 hours)).

[0089] Anti-beta-amyloid antibodies or fragments thereof may be selected from those known to those skilled in the art. Suitable antibodies may include human or humanized anti-Aβ monoclonal antibodies from the group consisting of bapineuzumab (Pfizer Inc. / Janssen Pharmaceuticals, Inc.), solanezumab (Eli Lilly and Company), gantenerumab (Hoffman-La Roche), crenezumab (Genentech, Inc.), ponezumab (Pfizer Inc.), donanemab (Eli Lilly), BAN2401 / lecanemab (Lecanamab) (BioArctic Neuroscience, AB / Eisai Co., Ltd / Biogen, Inc.), and aducanumab (Aduhelm™ Biogen, Inc.) (see also van Dyck CH., Biol Psychiatry 83(4):311-319, 2018). Suitable antibody fragments may include Fab fragments and scFv antibodies that target Aβ containing scFv molecules, as described by Sebollela A., J Neurochem 142(6):934-937, 2017 and Zha J et al., Scientific Reports 6:36631, 2016.

[0090] In some embodiments, the above method will be used to prevent, reduce the symptoms of, and / or slow the progression of AD or other diseases associated with pathogenic protein misfolding, aggregation, and deposition (including Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS) (i.e., Lou Gehrig's disease), Pick's disease, Gaucher's disease, and frontotemporal degeneration (FTD)). Therefore, the above method can, for example, prevent the onset, development, or progression of a disease or condition, or the onset, development, or progression of one or more symptoms or adverse features of a disease or condition (e.g., toxic aggregation of Aβ protein). Since beta-amyloid and tau aggregation is known to appear in the early stages of Alzheimer's disease, the present invention offers significant potential to enable therapeutic methods intended to prevent and / or remove toxic aggregates of Aβ or tau to be achieved by administering ambroxol (or related compounds) or analogs of the present invention.

[0091] In a modified form, the present invention may provide a method for preventing, reducing the symptoms of, and / or slowing the progression of AD or other diseases associated with pathogenic protein misfolding, aggregation, and deposition (including Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS) (i.e., Lou Gehrig's disease), Pick's disease, Gaucher's disease, and frontotemporal degeneration (FTD)), comprising the step of administering to a subject an effective amount of the present invention's ambroxol (or related compounds such as ambroxol hydrochloride and bromhexine) or analog. That is, the present invention's ambroxol or related compounds or analog may be used as the sole activator.

[0092] In some embodiments of methods for preventing, reducing the symptoms of, and / or slowing the progression of AD or other diseases associated with pathogenic protein misfolding, aggregation, and deposition (including Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS) (i.e., Lou Gehrig's disease), Pick's disease, Gaucher's disease, and frontotemporal degeneration (FTD)), subjects may be selected based on preferred biomarkers indicating patients at risk of or in the early stages of AD or other diseases associated with pathogenic protein misfolding, aggregation, and deposition (including Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS) (i.e., Lou Gehrig's disease), Pick's disease, Gaucher's disease, and frontotemporal degeneration (FTD)). In AD, biomarker levels may be found in CSF and / or blood in the early or preclinical stages of AD. For example, see Janelidze, S. et al., "Plasma P-tau181 in Alzheimer's Disease: Relationship to Other Biomarkers, Differential Diagnosis, Neuropathology and Longitudinal Progression to Alzheimer's Dementia," Nat Med 26:379-386; also see Hansson et al., "The Alzheimer's Association appropriate use recommendations for blood biomarkers in Alzheimer's disease," Alzheimer's & Dementia: 2669-2686 (2022).Therefore, in one example, the above method uses phosphorylated tau protein (p-tau) p-tau217 (i.e., tau phosphorylated at the Thr-217 residue) (Palmquvist et al., "Discriminative Accuracy of Plasma Phospho-tau217 for Alzheimer Disease vs Other Neurodegenerative Disorders" JAMA 324:772-781 (2020)) or p-tau181 (i.e., tau phosphorylated at the Thr-181 residue) (Janelidze et al., "Plasma P-tau181 In Alzheimer's Disease: Relationship To Other Biomarkers, Differential Diagnosis, Neuropathology And Longitudinal Progression To Alzheimer's Dementia," Nat Med 26:379-386 (2020)), or oligomerized species of amyloid-beta (Shea et al., "SOBA: Development and Testing The assay may further include the step of selecting subjects by assaying the increase in levels of a suitable sample (e.g., a sample of CSF, whole blood, or plasma) of "Of A Soluble Oligomer Binding Assay For Detection Of Amyloidogenic Toxic Oligomers" Proceedings of the National Academy of Sciences 119:e2213157119 (2022)).These biomarkers can accurately identify patients with no cognitive impairment, mild cognitive impairment, or memory impairment classified as having AD dementia, and have been shown to accurately predict patients who will progress to AD dementia (Janelidze et al., 2020; Palmqvist et al., "Prediction Of Future Alzheimer's Disease Dementia Using Plasma Phospho-Tau Combined With Other Accessible Measures," Nat Med 27:1034-1042 (2021); Shea et al., 2022; Jia J., et al. "Biomarker Changes during 20 Years Preceding Alzheimer's Disease," New England Journal of Medicine, Volume 390, No. 8, pp 712-722 (2024)). These same species may be monitored longitudinally in treated subjects to evaluate the effects of the treatment. Other p-tau isoforms, such as p-tau231 (Ashton, NJ et al. Acta Neuropathol 1-16 (2021)) or p-tau235 (Lantero-Rodriguez, J. et al. Embo Mol Med 13, e15098 (2021)), may also be assayed. Other potential biomarkers include glucose metabolism or aggregated proteins such as beta-amyloid or tau, as measured by positron emission tomography (PET) (Therriault, J. et al. Nat Aging 1-10 (2022) doi:10.1038 / s43587-022-00204-0. Iaccarino, L., et al. "Journal of Alzheimer's Disease, 59, pp 603-614 (2017)).Retinal imaging-based biomarkers, including amyloid deposition and thinning of the nerve fiber layer, can also be used for the early diagnosis of Alzheimer's disease (Snyder, PJ et al. Alzheimers Dement. Diagn. Assess. Dis. Monit. 4, 169-178 (2016); Koronyo, Y. et al. JCI Insight 2, (2017)).

[0093] Other suitable biomarkers indicating patients at risk of AD or other amyloidosis, or those in the early stages of the disease, include, for example, the ε4 allele of the ApoE gene, the apolipoprotein E prototype (C2N Diagnostics), pyroglutamate-modified amyloid-beta protein, N3pG, pTau217, and / or the amyloid-beta 42 / 40 ratio (Aβ42 / 40) (C2N Diagnostics). Plasma NFL (neurofilament light chain) is elevated in many neurodegenerative diseases. This is not specific to any particular neurodegenerative disease, but rather reflects disease activity and severity. For example, plasma NFL is elevated in patients with multiple sclerosis with actively progressing demyelination, but decreases when the patient is treated with immunosuppressants (Barro, C. & Zetterberg, H. Acta Neurol Scand (2021) doi:10.1111 / ane.13415). Plasma exosomes accompanied by neuroproteins also appear to be promising biomarkers for diagnosing several neurodegenerative diseases, including Parkinson's disease, Alzheimer's disease, and head injury (Rastogi, S. et al. Int J Mol Sci 22, 440 (2021)).

[0094] Examples of additional markers that have been used to identify patients at risk include, for example, Roche's Elecsys AD (Ab42, Ab40, p-tau181, and total tau) or Fujirebio's beta-amyloid ratio test (e.g., Ab42 and Ab40), Eli Lilly's Tauvid (flourtaucipir - detects tau on PET) or Amyvid (flourbetapir - detects amyloid on PET), or Neuraceq (fluorbetapen) or Vizamyl (flutametamol).

[0095] In some embodiments of methods for preventing, reducing the symptoms of, and / or slowing the progression of AD or other diseases associated with pathogenic protein misfolding, aggregation, and deposition (including Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS) (i.e., Lou Gehrig's disease), Pick's disease, Gaucher's disease, and frontotemporal degeneration (FTD)), subjects may be selected based on genotyping of at least one gene or locus that points to a patient at risk of AD or other diseases associated with pathogenic protein misfolding, aggregation, and deposition (including Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS) (i.e., Lou Gehrig's disease), Pick's disease, Gaucher's disease, and frontotemporal degeneration (FTD)). In Alzheimer's disease (AD), genotyping may be based on the gene encoding apolipoprotein E (ApoE); subjects carrying the ε4 allele of ApoE have an increased risk of AD compared to subjects carrying the more common ε3 allele, while those carrying the ε2 allele have a reduced risk (Liu CC et al., Nat Rev Neurol. 9(2):106-118, 2013). Preferred methodologies for performing ApoE genotyping are known to those skilled in the art and include, for example, preferred RT-PCR protocols (see, e.g., Zhong L et al., Mol Neurodegener. 11:2, 2016). However, in other embodiments, subjects may be selected based on genotyping of p-tau217, p-tau181, and / or N3pG.

[0096] In some embodiments of methods for preventing and / or treating Alzheimer's disease (AD) or other diseases associated with pathogenic protein misfolding, aggregation, and deposition (including Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS) (i.e., Lou Gehrig's disease), Pick's disease, Gaucher's disease, and frontotemporal degeneration (FTD)), ambroxol (or related compounds) or analogs of the present invention are used. (i) A dosage that provides a peak concentration in the subject's serum that is greater than 1 μM, for example, 2–50 μM, 2–25 μM, or 10–20 μM. (ii) A dosage that provides a peak concentration in the subject's brain tissue greater than 3 μM, for example, 5–50 μM, 5–25 μM, or 10–20 μM, or (iii) Dosage within the range of approximately 250 mg to 1000 mg / day or 750 to 1000 mg / day In some cases, it may be preferable to administer the drug to the subject at a relatively high daily dose selected from the available options. This is because such high daily doses may be necessary to allow the ambroxol (or related compound) or analog of the present invention to upregulate the secretion of toxic aggregated proteins (e.g., aggregated Aβ) from affected cells. Pharmaceutical composition

[0097] In some embodiments, the present invention includes a composition comprising a therapeutically effective amount of a compound of formula I (e.g., the compound shown in Figure 1) or a pharmaceutically acceptable salt, solvate, or prodrug thereof, in combination with a pharmaceutically acceptable carrier, for preventing and / or treating various diseases and medical conditions, including respiratory diseases and conditions, lysosomal storage disorders (LSD), and neurological diseases and conditions, or for extending the life expectancy of a subject. For example, a composition comprising a compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) may be used to (a) treat, inhibit, or reduce aging in a subject, (b) treat, inhibit, or reduce age-related symptoms or age-related diseases in a subject, and / or (c) increase the healthy lifespan, life expectancy, and / or intellectual sharpness of a subject.

[0098] The compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) may be administered in combination with a pharmaceutically acceptable carrier in the form of a pharmaceutical composition. In such a composition, the compound of formula I may constitute 0.1 to 99.99 percent by weight.

[0099] The compound of formula I is preferably administered with a pharmaceutically acceptable carrier selected based on a chosen route of administration and standard pharmaceutical practice. The compound of formula I may be formulated into dosage forms in accordance with standard practice in the field of pharmaceutical preparations. Alphonso Gennaro, ed., Remington's Pharmaceutical Sciences, 18 th See Edition (1990), Mack Publishing Co., Easton, Pa. Preferred dosage forms may include, for example, tablets, capsules, liquids, parenteral liquids, lozenges, suppositories, or suspensions. Preferred examples of preparations of oral, topical, suppository, and parenteral formulations of ambroxol, bromhexine, or other ambroxol derivatives readily adaptable to the compounds of the present invention are disclosed, for example, in Examples 1-8 of WO2005 / 007146 or its equivalent US2005 / 00148747, which are incorporated herein by reference.

[0100] In another aspect, the present invention provides the use of compounds of formula I in the preparation of pharmaceuticals for preventing and / or treating various diseases and medical conditions, including respiratory diseases and conditions, lysosomal storage disorders (LSDs), and neurological diseases and conditions, or for extending the life expectancy of subjects.

[0101] For parenteral administration, the compound of formula I may be mixed with a suitable carrier or diluent, for example, water, oil (especially vegetable oil), ethanol, physiological saline, aqueous dextrose (glucose) and related sugar solutions, glycerol, or glycols such as propylene glycol or polyethylene glycol, or with plant extracts as supplements. The parenteral administration solution preferably contains a water-soluble salt of the activator. Stabilizers, antioxidants, and preservatives may be added. Suitable antioxidants include sulfites, ascorbic acid, citric acid and its salts, and sodium EDTA. Suitable preservatives include benzalkonium chloride, methylparaben or propylparaben, and chlorobutanol. The parenteral administration composition may take the form of an aqueous or non-aqueous solution, dispersion, suspension, or emulsion.

[0102] For oral administration, the compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) may be combined with one or more solid inert components for the preparation of tablets, capsules, pills, powders, granules, or other suitable oral dosage forms. For example, the compound of formula I may be combined with at least one excipient, e.g., a filler, binder, humectant, disintegrant, dissolution retarder, absorption accelerator, wetting agent, absorbent, or lubricant. According to one tablet embodiment, the compound of formula I may be combined with carboxymethylcellulose calcium, magnesium stearate, mannitol, and starch, and then formed into tablets by a conventional tableting method.

[0103] For oral administration, the compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) may be provided in a liquid oral pharmaceutical composition. Liquid oral dosage forms can offer unique advantages over solid dosage forms such as tablets and capsules. For example, in many cases, the amount of active ingredient required to treat a particular disease or condition necessitates multiple tablets or capsules once or multiple times a day. Taking multiple tablets or capsules involves not only the active pharmaceutical ingredient but also multiple doses of excipients used to formulate the tablets and capsules. Such excipients, such as oils and alcohols, are typically not well tolerated by many patients and generally lead to gastric discomfort. In addition, liquid oral dosage forms are easier for patients to adhere to than solid dosage forms because the delivery of the active pharmaceutical ingredient is achieved with only one or two doses per day. Furthermore, liquid oral dosage forms provide rapid absorption of the active pharmaceutical ingredient from the gastrointestinal tract. Moreover, liquid oral dosage forms allow for the use of flavorings and / or palatability agents, thereby further promoting patient acceptance and adherence to medication.

[0104] Therefore, in some preferred embodiments, the compositions of the present invention are liquid oral pharmaceutical compositions. Such compositions may be particularly suitable for preventing and / or treating lysosomal storage disorders (LSDs) such as Gaucher disease, or neurological disorders or conditions such as Parkinson's disease (PD). In some embodiments, the compositions will contain a “highly loading” compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) to deliver an effective amount in a small volume (e.g., a 15 mL dose, once or twice a day or more). Highly drug-loading liquid oral pharmaceutical compositions according to the present invention may offer at least one of the following advantages: (1) improved absorption in the gastrointestinal tract; (2) maintenance of effective blood concentration over 24 hours; (3) reduced undesirable side effects of excipients compared to solid dosage forms; (4) reduced number of doses required; and (5) improved taste and mouthfeel quality.

[0105] In one specific embodiment of the high-drug-loading liquid oral pharmaceutical composition of the present invention, the composition comprises (i) a compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) and (ii) at least one pharmaceutically acceptable excipient, wherein the compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) is in the form of a granule having a granule core containing about 60 to about 97 weight percent of the active pharmaceutical ingredient and about 3 to about 40 weight percent of the excipient, where the weight percentage is based on the total weight of the granule core.

[0106] In another specific embodiment of the high-drug-loading liquid oral pharmaceutical composition of the present invention, the composition comprises (i) a compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof), (ii) at least one pharmaceutically acceptable excipient, and (iii) a diluent, wherein the compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) is in the form of a granule having a granule core containing about 60 to about 97 weight percent of the active pharmaceutical ingredient and about 3 to about 40 weight percent of the excipient, where the weight percentage is based on the total weight of the granule core, wherein the granule core is coated with (iv) a water-soluble seal coating in an amount that provides a weight increase of about 0.5 to about 5 percent, and (v) an enteric coating in an amount that provides a weight increase of about 0.5 to about 50 percent.

[0107] Such a high drug-loading liquid oral pharmaceutical composition may be prepared by a method comprising, for example, the steps of: (a) preparing a granule having a granule core comprising about 60 to about 97 weight percent of a compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) and about 3 to about 40 weight percent of at least one pharmaceutically acceptable excipient, wherein the weight percent is based on the total weight of the granule core; (b) coating the granule with a water-soluble seal coating in an amount that provides a weight increase of about 0.5 to about 5 percent; (c) coating the granule prepared in step (b) with an enteric coating in an amount that provides a weight increase of about 0.5 to about 50 percent; and (d) preparing a liquid suspension comprising the enteric-coated granule prepared in step (c) and the liquid suspension formulation, wherein the liquid suspension formulation comprises a suspending agent, a vehicle for enhancing the stability of the high drug-loading liquid oral pharmaceutical composition, and a diluent.

[0108] The compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) is preferably in the form of granules, having a particle size of about 100 to about 500 microns, more preferably about 100 to about 300 microns, about 150 to about 350 microns, and about 200 to about 350 microns after micronization; and even more preferably, the particle size is about 100 to about 200 microns, about 150 to about 250 microns, about 200 to about 300 microns, about 200 to about 400 microns, about 250 to about 350 microns, about 250 to about 450 microns, about 300 to about 400 microns, about 300 to about 500 microns, about 350 to about 450 microns, and / or about 400 to about 500 microns. Particle sizes within these ranges are known to be coatable and provide ease of swallowing without prompting the user to chew. For example, in preferred embodiments, particle sizes from about 250 to about 350 microns have been found to balance the ability to coat with the ability to avoid a sandy "mouthfeel." Particle size can be determined, for example, by laser light scattering using a Malvern mastersizer device MS2000 equipped with a hydro-S dispersion unit. Micronization of the compound of formula I may be carried out in a dry environment using dry mills such as cutting mills, pin / cage mills, hammer mills, jet mills, fluidized bed jet mills, and ball mills.

[0109] A preferred excipient that can be used to prepare the granular core is a binder. The binder may be any water-soluble, pharmaceutically acceptable polymer. In one embodiment, the compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) is in the form of a powder, which the binder necessarily binds due to the poor tackiness of most powders. Preferably, the binder is selected from povidone (polyvinylpyrrolidone), copovidone (vinylpyrrolidone-vinyl acetate copolymer), microcrystalline cellulose, powdered cellulose, crystalline cellulose, silicon-treated microcrystalline cellulose, cellulose derivatives, e.g., hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, and hydroxypropylmethylcellulose, starch, pregelatinized starch, polymethacrylate, compressible sugars, sucrose, and sugar alcohols, e.g., mannitol, sorbitol, maltitol, and xylitol, and mixtures thereof. More preferably, the binder is hydroxypropylcellulose (Klucel LF).

[0110] Granules used in high-load liquid oral pharmaceutical compositions are prepared by direct spheroidization, which involves preparing a granule core containing, preferably, about 60 to about 97 weight percent, preferably about 75 to about 97 weight percent, and more preferably more than about 90 to 97 weight percent of a compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) and about 3 to about 40 weight percent, preferably about 3 to about 25 weight percent, and more preferably about 3 to about 10 weight percent of a binder, where weight percent is based on the total weight of the granule core. Granulation may be carried out under high shear (mixer granulation) or in a fluidized bed (fluidized bed granulation).

[0111] The granular core is sealed as needed with a water-soluble seal coating in an amount that provides a weight increase of approximately 0.5 to 5 percent, 0.5 to 3 percent, 0.5 to 2 percent, 0.5 to 1 percent, 1 to 2 percent, 1 to 3 percent, 1 to 4 percent, 2 to 3 percent, 2 to 4 percent, 1 percent or more, 2 percent or more, 3 percent or more, or 4 percent or more. It has been found that applying a water-soluble seal coating produces smoother and more uniform granules that are more receptive to enteric coating. Preferred water-soluble polymers include hydroxypropyl methylcellulose (HPMC), carboxymethylcellulose (sodium and calcium salts), ethylcellulose, methylcellulose, hydroxyethylcellulose, ethylhydroxyethylcellulose, hydroxypropylcellulose (HPC), L-HPC (low-substituted HPC), polyvinylpyrrolidone, polyvinyl alcohol, polymers of acrylic acid and its salts, vinylpyrrolidone-vinyl acetate copolymers (e.g., Kollidon® VA64, BASF), gelatin, guar gum, partially hydrolyzed starch, alginates, and xanthan gums. Most preferably, the water-soluble polymer is hydroxypropyl methylcellulose. The seal coating is preferably applied using a bottom-spray fluidized bed coater equipped with a Wolster column.

[0112] In addition to or in the absence of a seal coating, the granular core may be optionally coated with an enteric coating directly on the granular core or on a seal coating pre-applied to the granular core. Preferably, the enteric coating is applied in an amount that provides a weight increase of about 0.5 to about 50 percent, about 1 to about 40 percent, about 2 to about 30 percent, about 3 to about 20 percent, or about 4 to about 10 percent, based on the total weight of the granular core. More preferably, the enteric coating is applied in an amount that provides a weight increase of about 0.5 to about 5 percent, about 1 to about 4 percent, or about 2 to about 3 percent, based on the total weight of the granular core. The enteric coating is preferably applied using a bottom-spray fluidized bed coater equipped with a Wurster column.

[0113] Enteric coatings include polymers selected from acrylate polymers or aqueous cellulose dispersions. Combinations of acrylate polymers and / or aqueous cellulose dispersions may also be used. Preferably, the acrylate polymer is selected from polymethacrylate methyl methacrylate copolymer (Eudragit® L-100), polyethyl acrylate methyl methacrylate trimethylammonium ethyl methacrylate chloride copolymer (Eudragit RL-100, RS-100), polymethacrylate ethyl acrylate copolymer (Eudragit® L30D-55), ethyl acrylate methyl methacrylate trimethylammonium ethyl methacrylate chloride copolymer (Eudragit® RL30D), ethyl acrylate methyl methacrylate trimethylammonium ethyl methacrylate chloride copolymer (Eudragit® RS30D), and polyethyl acrylate methyl methacrylate copolymer (Eudragit® NE30D). More preferably, the acrylate polymer is polymethacrylate ethyl acrylate copolymer (Eudragit® L30D-55). The enteric coating may preferably be applied using a bottom-spray fluid bed coater equipped with a Wolster column. The enteric coating can increase the delivery of the active pharmaceutical ingredient to the gastrointestinal tract region of the subject where the pH is between about 4.5 and about 6.5. The enteric coating also increases the delivery of the compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) to the proximal or central part of the small intestine or both. In addition, the enteric coating can increase the delivery of the compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) to one or more of the central parts of the duodenum, jejunum, or ileum. Preferably, the enteric coating begins to dissolve in an aqueous solution with a pH between about 4.5 and about 5.5.

[0114] The vehicle for enhancing the stability of the high-drug-loading liquid oral pharmaceutical composition may preferably be a protective colloid.

[0115] The high-load liquid oral pharmaceutical composition of the present invention may contain additional plasticizers. Preferred plasticizers are diethyl phthalate, dibutyl phthalate, triethyl citrate, and glycerol. A combination of plasticizers may also be used. More preferably, the plasticizer is triethyl citrate.

[0116] In one embodiment, the high-drug-load liquid oral pharmaceutical composition is in the form of a solution. In another embodiment, the high-drug-load liquid oral pharmaceutical composition is in the form of a suspension. Enteric-coated granules may be combined with the liquid suspension formulation to form a suspension. The liquid suspension formulation may include a suspending agent, a vehicle for enhancing the stability of the high-drug-load liquid oral pharmaceutical composition, and a diluent. Since many suspending agents also act as stability enhancers, the suspending agent and the vehicle for enhancing the stability of the high-drug-load liquid oral pharmaceutical composition may be the same or different.

[0117] Examples of suspending agents include, but are not limited to, methylcellulose, sodium carboxymethylcellulose, hydroxyethylcellulose, hydroxypropylmethylcellulose, tragacanth gum, and glycerol monostearate. More preferably, the vehicle for enhancing the stability of the liquid oral pharmaceutical composition is a protective colloid. Examples of protective colloids include, but are not limited to, hydroxymethylcellulose, sodium carboxymethylcellulose, polyvinyl alcohol, gelatin, and polyvinylpyrrolidone. Combinations of protective colloids may be used. Preferred protective colloids are mixtures of cellulose gum, xanthan gum, and carrageenan. Examples of diluents include, but are not limited to, water, alcohol (e.g., methyl alcohol, ethyl alcohol, propyl alcohol, i-propyl alcohol, etc.), acetone, glycerin, oil (e.g., castor oil, etc.), any other pharmaceutically acceptable diluent, or mixtures thereof. Most preferably, water is used as the suspension or solution diluent. In addition, pH modifiers and / or antioxidants may be used.

[0118] In some specific embodiments of the liquid suspension formulations of the present invention, the formulation comprises microcrystalline cellulose and sodium carboxymethylcellulose (Avicel RC-591), gum (CP Kelco), and water; or microcrystalline cellulose and sodium carboxymethylcellulose (Avicel RC-591), cellulose gum, xanthan gum, and carrageenan (Ticaloid Ultrasmooth), and water.

[0119] According to a preferred embodiment, the liquid suspension formulation comprises at least one suspending agent in an amount of about 0.5% to about 3%, at least one protective colloid in an amount of about 0.5% to about 1%, and about 98% of a diluent, where the weight percentage values ​​are based on the weight of the liquid suspension formulation. More preferably, the protective colloid will be used in an amount of about 0.1 to 0.5 weight percent. In some specific embodiments, the liquid suspension formulation comprises at least one suspending agent in an amount of about 1.5%, at least one protective colloid in an amount of about 0.2% or more, and about 98% of a diluent.

[0120] Preferably, the high-drug-loading liquid oral pharmaceutical composition in suspension form has a viscosity of less than about 5 Pa₂s. More preferably, the high-drug-loading liquid oral pharmaceutical composition in suspension form has a viscosity of less than about 3 Pa₂s, most preferably less than about 1 Pa₂s.

[0121] In some embodiments, high-drug-load liquid oral pharmaceutical compositions are used to prevent and / or treat lysosomal storage disorders selected from Gaucher disease (including types 1, 2, and 3 Gaucher diseases), Pompe disease (including infantile and late-onset forms), and Fabry disease, or to prevent and / or treat Parkinson's disease (e.g., to alleviate its symptoms). See, for example, Lukas J. et al. "Enzyme enhancers for the treatment of Fabry and Pompe disease.." Mol Ther. 2015 Mar;23(3):456-64.

[0122] In some more specific embodiments, a high-drug-loading liquid oral pharmaceutical composition is used to prevent and / or treat AD, or other diseases associated with pathogenic protein misfolding, aggregation, and deposition (including Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS) (i.e., Lou Gehrig's disease), Pick's disease, Gaucher disease, and frontotemporal degeneration (FTD)). In some further specific embodiments, a high-drug-loading liquid oral pharmaceutical composition is used to prevent and / or treat subjects with mutations in glucocerebrosidase (GC-ase) (e.g., mutations in beta-glucocerebrosidase, where the subject may also have Gaucher disease, AD, and / or Parkinson's disease). Mutations in beta-glucocerebrosidase are a. The following locations: D140H, V15L, G46E, K79N, R119Q, P122S, R131L, K157Q, N188S, Y212H, F213I, F216V, F216Y, H225Q, F251L, R257E, P289L, A309V, H311R, W312C, Y323I, G325R, E326K, C342G, R Point mutations in any one of the following: 353G, R359X (terminal), S364T, N370S, L371V, G377S, V394L, V398F, P401L, D409H, D409V, P415R, L444P, A456P, V460V, R463C, G478S, or R496H, and / or any combination thereof. b. Point mutation in L444P, c. Point mutation in N370S, d. Point mutation in E326K, e. Point mutations in L444P, A456P, and V460V. f. Point mutations in D140H and E326K, g. Point mutations in H255Q and D409H, h. Guanine insertion in 84GG, i. Splice site mutations in intron 2 (IVS2DS+lG-A) that result in exon 2 skipping, j. 1-bp deletion in the GCase gene (1023delC in the genome sequence), k. 55-bp deletion in the GC-ase gene (nucleotides 5879-5933 in genomic DNA), l. Homozygous 259C-T transition (1763 in genomic DNA), m. Homozygous 1-bp deletion in the GCase gene, resulting in a protein frameshift and premature cleavage at exon 6, and n. A G-to-A substitution at the first position within the splice site of intron 10 of the GCase gene, resulting in the insertion of the first 11 base pairs of IVS10 and the deletion of the first 11 base pairs of exon 11. You may choose from the following.

[0123] In some embodiments, the high-drug-load oral liquid pharmaceutical composition may be administered to subjects also receiving enzyme replacement therapy (e.g., combination therapy). Examples of such enzyme replacement therapies include, but are not limited to, recombinant glucocerebrosidases such as imiglucerase, veraglucerase, taliglucerase alfa (ELELYSO®), and / or eliglustat (CERDELGA®). The high-drug-load oral liquid pharmaceutical composition may be administered concurrently with enzyme replacement therapy, sequentially, or at different times.

[0124] In some embodiments, the high-drug-loading oral liquid pharmaceutical composition comprises an analog of ambroxol or bromhexine according to Formula I, or a pharmaceutically acceptable salt thereof, for treating subjects having misfolded and / or incorrectly transported glucocerebrosidase. In further embodiments, the high-drug-loading oral liquid pharmaceutical composition comprises an analog of ambroxol or bromhexine according to Formula I, or a pharmaceutically acceptable salt thereof, for treating or preventing lysosomal storage disorders in subjects. In yet another embodiment, the high-drug-loading oral liquid pharmaceutical composition comprises an analog of ambroxol or bromhexine according to Formula I, or a pharmaceutically acceptable salt thereof, for treating subjects having mutations in glucocerebrosidase. In yet another embodiment, the high-drug-loading oral liquid pharmaceutical composition comprises an analog of ambroxol or bromhexine according to Formula I, or a pharmaceutically acceptable salt thereof, for treating subjects having mutations in beta-glucocerebrosidase. In some preferred embodiments, the mutation in beta-glucocerebrosidase is selected from N370S, L444P, and / or E326K. In further embodiments, the high-drug-load oral liquid pharmaceutical composition comprises an analog of ambroxol or bromhexine according to formula I, or a pharmaceutically acceptable salt thereof, for treating a subject suffering from Gaucher disease. In further embodiments, the high-drug-load oral liquid pharmaceutical composition comprises an analog of ambroxol or bromhexine according to formula I, or a pharmaceutically acceptable salt thereof, for treating a subject suffering from PD.

[0125] The pharmaceutical compositions of the present invention may be formulated in unit dosage forms, each dose containing about 50 to about 1000 mg, more typically about 250 to about 500 mg, of the compound of formula I per unit dose. The term “unit dosage form” refers to a physically discontinuous unit suitable as a unit dose for human subjects and other mammals, in which each unit contains a predetermined amount of the compound of formula I, calculated to produce a desired therapeutic effect, associated with a suitable pharmaceutically acceptable carrier.

[0126] In a further preferred embodiment, the compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) is administered in several doses over a given period, for example, in a daily dose over a week or longer. For example, daily doses of approximately 20–500 mg / day, 50–150 mg / day, 50–200 mg / day, 50–250 mg / day, 250–500 mg / day, 250–1000 mg / day, 1000–1500 mg / day, 1500–2000 mg / day, or up to 1000–2000 mg / day may be available. Higher or lower doses are also intended, as it may be necessary to use doses outside these ranges in some cases. The daily dose may be divided, for example, into two to four daily doses.

[0127] The pharmaceutical compositions of the present invention may be formulated to provide slow or controlled release of the active ingredient, for example, by using a variable proportion of hydroxypropyl methylcellulose, other polymer matrices, gels, permeable membranes, penetration systems, multilayer coatings, microparticles, liposomes and / or microspheres to provide a desired release profile.

[0128] Generally, controlled-release formulations are pharmaceutical compositions that can release the active ingredient at the rate required to maintain a certain level of pharmacological activity over a desired period of time. Such dosage forms provide a supply of the drug to the body over a predetermined period and therefore maintain drug levels within the therapeutic range for a longer period than conventional uncontrolled formulations.

[0129] U.S. Patent No. 5,674,533 discloses a controlled-release pharmaceutical composition in liquid dosage form for the administration of mogistein, a potent peripheral cough suppressant. U.S. Patent No. 5,059,595 describes the controlled release of an activator using a gastric-tolerant tablet for the therapy of organic mental disorder. U.S. Patent No. 5,591,767 describes a liquid reservoir transdermal patch for the controlled administration of ketorolac, a nonsteroidal anti-inflammatory drug with potent analgesic properties. U.S. Patent No. 5,120,548 discloses a controlled-release drug delivery device composed of a swellable polymer. U.S. Patent No. 5,073,543 describes a controlled-release formulation containing a trophic factor encapsulated by a ganglioside-liposome vehicle. U.S. Patent No. 5,639,476 discloses a stable solid controlled-release formulation having a coating derived from an aqueous dispersion of a hydrophobic acrylic polymer. Biodegradable microparticles are known for their use in controlled-release formulations. U.S. Patent No. 5,354,566 discloses a controlled-release powder containing an active ingredient. U.S. Patent No. 5,733,566 describes the use of polymeric microparticles for releasing an antiparasitic composition. Any or all of these techniques may be adapted for the controlled release of a compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof).

[0130] The controlled release of a compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) may be stimulated by various inducers, such as pH, temperature, enzymes, water, or other physiological conditions or compounds. Various mechanisms of drug release exist. For example, in one embodiment, the controlled-release component may swell to form a porous opening large enough to release the active ingredient after administration to a patient. In the context of this invention, the term “controlled-release component” is defined herein as one or more compounds, such as polymers, polymer matrices, gels, permeable membranes, liposomes, and / or microspheres, that facilitate the controlled release of a compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) in a pharmaceutical composition. In another embodiment, the controlled-release component is biodegradable and induced by exposure to an aquatic environment in the body, pH, temperature, or enzymes. In another embodiment, a sol-gel may be used, in which the active ingredient is incorporated into a sol-gel matrix that is solid at room temperature. This matrix is ​​implanted in a subject, preferably a human or other mammal, that has a sufficiently high body temperature to induce gel formation of the sol-gel matrix, thereby releasing the active ingredients into the subject.

[0131] Compositions containing the compounds of the present invention that are suitable for intranasal or inhalation administration are of particular interest. For this reason, the compounds of formula I may be formulated for intranasal or inhalation administration typically in the form of a dry powder from a dry powder inhaler (either alone or in a mixture, for example, as a dry mixture with lactose in anhydrous or monohydrate form, preferably monohydrate, mannitol, dextran, glucose, maltose, sorbitol, xylitol, fructose, sucrose, or trehalose, or as mixed component particles mixed with phospholipids, for example), or as an aerosol spray from a pressurized vessel, pump, spray, atomizer (preferably an atomizer that uses electrohydrodynamics to produce a misty atomizer), or nebulizer, with or without a suitable propellant such as dichlorofluoromethane.

[0132] A pressurized vessel, pump, spray, atomizer, or nebulizer contains a solution or suspension of a compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof), for example, ethanol (or aqueous ethanol if necessary) or a suitable alternative for dispersing, solubilizing, or extending the release of the compound, a propellant as a solvent, and a surfactant as needed, such as sorbitan trioleate or oligolactic acid.

[0133] Before use in dry powder or suspension formulations, the compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) is pulverized to a size suitable for inhalation delivery (typically less than 5 microns). This may be achieved by any suitable crushing method, such as spiral jet milling, fluidized bed jet milling, supercritical fluid treatment to form nanoparticles, high-pressure homogenization, or spray drying.

[0134] Suitable solution formulations of the compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) for use in sprayers that generate a mist using electrohydrodynamics may contain 1 μg to 20 mg of the compound per operation, and the working volume may vary from 1 μL to 100 μL. Typical formulations may contain the compound of formula I, propylene glycol, sterile water, ethanol, and sodium chloride. Alternative solvents that can be used instead of propylene glycol include glycerol and polyethylene glycol. Capsules, blisters, and cartridges (e.g., made of gelatin or HPMC) for use in inhalers or injectors may be formulated to contain a mixed powder of the compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof), a suitable powder base such as lactose or starch, and a performance modifier such as L-leucine, mannitol, or magnesium stearate.

[0135] Formulations for inhalation / intranasal administration of compounds of formula I (or pharmaceutically acceptable salts, solvates, or prodrugs thereof) may be formulated to have immediate and / or controlled release. Controlled release formulations include delayed, sustained, pulsed, controlled dual, targeted, and programmed release formulations. Sustained or controlled release may be obtained, for example, by using poly(D,L-lactic acid-co-glycolic acid). Administration of the compound of the present invention

[0136] In some preferred embodiments, the compounds of the present invention are administered orally to the patient. However, the compounds may be administered by any route, including rectal, pulmonary, sublingual, and parenteral administration. Parenteral administration includes, for example, intravenous, intramuscular, intra-arterial, intraperitoneal, intranasal, intravaginal, intrasacral (e.g., into the bladder), intradermal, transdermal, topical, or subcutaneous administration.

[0137] The interval between doses may be once a week, twice a week, every other day, once a day, typically once, twice, three or four times a day, and the dose is given at equal intervals throughout the day and night to maintain a constant presence of the drug. However, those skilled in the art will notice that the treatment schedule can be optimized for any given subject, and that administration of the compound of formula I (or a pharmaceutically acceptable salt, solvate or prodrug thereof) may occur at a frequency less than once a day. Treatment may be carried out for as long as necessary.

[0138] The specific dose of the compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) required to induce a therapeutically beneficial response in a subject will, of course, be determined by the specific circumstances of each individual subject, including the subject's size, weight, age, and sex, as well as the route of administration of the compound.

[0139] For example, daily dosages of approximately 20-500 mg / day, 50-150 mg / day, 50-200 mg / day, 50-250 mg / day, 250-500 mg / day, 250-1000 mg / day, 1000-1500 mg / day, or up to 1000-2000 mg / day may be used. However, higher or lower doses are also considered, as it may be necessary to use dosages outside these ranges in some cases. Daily dosages may be divided, for example, into two to four equal daily doses. Furthermore, since long-term administration may be required in a manner that induces, increases, and / or improves the subject's healthy lifespan, life expectancy, and / or intellectual sharpness, the compound is preferably administered in doses of approximately 50 mg / day, 75 mg / day, 100 mg / day, 150 mg / day, 200 mg / day, 250 mg / day, 300 mg / day, 350 mg / day, 400 mg / day, 450 mg / day, 500 mg / day, 550 mg / day, 600 mg / day, 650 mg / day, 700 mg / day, 750 mg / day, 800 mg / day, 850 mg / day, 900 mg / day, 950 mg / day, 1000 mg / day, 1050 mg / day, 1100 mg / day, and 1150 mg / day. It may be administered in doses of 1200 mg / day, 1250 mg / day, 1300 mg / day, 1350 mg / day, or between 50-150 mg / day, 50-200 mg / day, 50-250 mg / day, 250-500 mg / day, 250-1000 mg / day, or between 1000-1500 mg / day, or less than 1000 mg / day, or approximately between 1 mg / kg / day, 2 mg / kg / day, 3 mg / kg / day, 4 mg / kg / day, 5 mg / kg / day, 6 mg / kg / day, 7 mg / kg / day, 8 mg / kg / day, 9 mg / kg / day, 10 mg / kg / day, 11 mg / kg / day, 12 mg / kg / day, and / or between 4 and 12 mg / kg / day.

[0140] The present invention also provides a pharmaceutical pack or kit comprising one or more containers filled with one or more of the raw materials of the pharmaceutical composition of the present invention. As may be associated with such containers, a warning in the form prescribed by the government agency regulating the manufacture, use or sale of the pharmaceutical or biological product, the warning reflects the approval by the authority for manufacture, use or sale for human administration.

[0141] Without further description, those skilled in the art will find it easy to prepare and utilize compounds of formula I (e.g., the compounds shown in Figure 1) or pharmaceutically acceptable salts, solvates, or prodrugs thereof, as well as practical methods of the present invention, using the preceding descriptions and the following illustrative examples. The following examples are some of the preferred embodiments of the present invention, but should not be construed as limiting the present disclosure in any way. Furthermore, while the invention of this specification has been described with reference to embodiments, it should be understood that such embodiments and examples provided herein merely illustrate the principles and applications of the present invention. Accordingly, it should be understood that numerous modifications may be made to the illustrative embodiments and examples, and that other arrangements may be devised without departing from the spirit and scope of the invention as defined, for example, by the following claims. All patent applications, patents, documents, and references listed herein are thus incorporated herein by reference in their entirety. [Examples]

[0142] The present invention is further illustrated here by the following examples. It will be noted that these are merely examples and that modifications to the details are possible, while still remaining within the scope of the present invention. Example 1 Synthesis of ZW-010

[0143] The compounds shown in Figure 1 can be synthesized by adapting one or more synthetic schemes described in Latli B et al., J Label Compd Radiopharm 53:15-23, 2010, and / or the following synthetic methods for the synthesis of the difluoro and dimethoxy analogs of ambroxol, ZW-010, and ZW-011, respectively, and / or any other methods known to those skilled in the art. Synthesis of compound ZW-010 [ka] 1. Synthesis of (2-amino-3,5-difluorophenyl)methanol

[0144] Under nitrogen, a chilled (0°C) solution of 2-amino-3,5-difluorobenzoic acid (500 mg, 2.89 mmol) in anhydrous tetrahydrofuran (THF, 15 ml) was added dropwise with 2.54 ml, 6.11 mmol, 2.4 M lithium aluminum hydride solution in THF. After the addition was complete, the reaction mixture was stirred at ambient temperature for 1 hour, then cooled to 0°C and quenched with 2.0 N aqueous sodium hydroxide (NaOH). The aqueous mixture was filtered through Celite, and the filtrate was extracted with two 50 mL doses of dichloromethane. The combined organic extracts were dried (Na2SO4), filtered, and concentrated to obtain an orange crystalline solid, which was used without further purification (302 mg, 66%). MS(APCI): m / z160. 2. Synthesis of 2-amino-3,5-difluorobenzaldehyde

[0145] A solution of (2-amino-3,5-difluorophenyl)methanol (302 mg, 1.90 mmol) in toluene (15 mL) was treated with manganese(IV) oxide (1.50 g, 15.20 mmol, 88%), and the reaction mixture was stirred at 80°C for 30 minutes. After cooling to ambient temperature, excess reagent was filtered off (using a Celite pad), and the filtrate was concentrated. Elution by passing the filtrate through a small plug of flash silica gel (silica gel 60, 230-400 mesh, 7:3 v / v hexane:ethyl acetate (siRNA)) yielded the product as a yellow oily substance, which crystallized upon standing (143 mg, 48%). MS (APCI): m / z 158. 3. Synthesis of trans-4-{[(2-amino-3,5-difluorophenyl)methyl]amino}cyclohexanol, ZW-010, as a hydrochloride salt.

[0146] A mixture of 2-amino-3,5-difluorobenzaldehyde (140 mg, 0.89 mmol) and trans-4-aminocyclohexanol (103 mg, 0.891 mmol) in ethanol (15 mL) was heated at 80°C for 5 hours and then cooled to ambient temperature. Sodium borohydride (81 mg, 2.14 mmol) was then added, and the mixture was stirred at ambient temperature for 2 hours, followed by quenching with an aqueous solution of saturated ammonium chloride (NH4Cl). The mixture was basicized by the addition of a saturated sodium bicarbonate solution, and the product was extracted with ethyl acetate. The organic extract was dried (Na2SO4), filtered, and concentrated. A clear oily substance was obtained by elution through a flash chromatography column (silica gel 60, 230-400 mesh, 1:9 v / v methanol:ethyl acetate) to the correct mass [MS(APCI): m / z257]. The free base was dissolved in ethyl acetate, and the solution was treated dropwise with a 5-6 N HCl solution in isopropanol to precipitate the hydrochloride as an off-white solid. The precipitate was filtered, washed with ethyl acetate, and dried under vacuum at 35°C to obtain pure hydrochloride of ZW-010 (180 mg, 69%). MS(ESI): m / z 257.1(M+H) +;1H NMR (MeOD-d4) δ (ppm): 1.35 (m, 2H), 1.60 (m, 2H), 2.1 (m, 2H), 2.25 (m, 2H), 3.25 (m, 1H), 3.6 (m, 1H), 4.3 (s, 2H), 7.1 (m, 2H) Example 2 Synthesis of compound ZW-011 [ka] 1. Synthesis of (2-amino-3,5-dimethoxy)methanol

[0147] Under nitrogen, a chilled (0°C) solution of 2-amino-3,5-dimethoxybenzoic acid (1.0 g, 5.07 mmol) in anhydrous THF (30 mL) was added dropwise with 2.4 M lithium aluminum hydride solution (4.44 mL, 10.65 mmol) in THF. After the addition was complete, the reaction mixture was stirred at ambient temperature for 1 hour, recooled to 0°C, and quenched with 2.0 N NaOH. The aqueous mixture was filtered through Celite, and the filtrate was extracted with two 50 mL portions of dichloromethane. The combined organic extracts were dried (Na₂SO₄), filtered, and concentrated. The product was obtained as an orange crystalline solid (870 mg, 94%) by elution through a flash column (silica gel 60, 230-400 mesh, 7% MeOH in siRNA). MS (APCI): m / z 184. 2. Synthesis of 2-amino-3,5-dimethoxybenzaldehyde

[0148] A solution of (2-amino-3,5-dimethoxy)methanol (870 mg, 4.75 mmol) in dichloromethane (40 mL) was treated with manganese(IV) oxide (3.75 g, 38.00 mmol, 88%), and the reaction mixture was stirred at ambient temperature for 1.5 hours. The excess reagent was then filtered off (on a Celite pad), and the filtrate was concentrated. Elution by flash column (silica gel 60, 230-400 mesh, 1:1 v / v hexane:siRNA) yielded 2-amino-3,5-dimethoxybenzaldehyde as a yellow oily substance (270 mg, 31%). MS (APCI): m / z 182. 3. Synthesis of trans-4-{[(2-amino-3,5-dimethoxyphenyl)methyl]amino}cyclohexanol, ZW-011, as a hydrochloride salt.

[0149] A mixture of 2-amino-3,5-dimethoxybenzaldehyde (270 mg, 1.49 mmol) and trans-4-aminocyclohexanol (172 mg, 1.49 mmol) in ethanol (25 mL) was heated at 80°C for 4 hours and then cooled to ambient temperature. Sodium borohydride (135 mg, 3.58 mmol) was then added, and the mixture was stirred at ambient temperature for 1 hour, followed by quenching with a saturated aqueous solution of ammonium chloride (NH4Cl). The mixture was basicized with a saturated aqueous solution of NaHCO3 and extracted with ethyl acetate. The organic extract was dried (Na2SO4), filtered, and concentrated. Elution through a flash column (silica gel 60, 230-400 mesh, 45% MeOH in ethyl acetate) yielded an orange oily substance (MS(APCI): m / z 281). The free base was dissolved in HCl, and the solution was treated dropwise with a 5-6 N HCl solution in isopropanol to precipitate the hydrochloride salt of ZW-011 as a pale green amorphous solid. This was filtered, washed, and dried under vacuum at 35°C (207 mg, 44%). MS(ESI): m / z281.0(M+H)+; 1H NMR (MeOD-d4) δ (ppm): 1.35 (m, 2H), 1.60 (m, 2H), 2.1 (m, 2H), 2.25 (m, 2H), 3.3 (m, 1H), 3.6 (m, 1H), 3.9 (s, 3H), 4.0 (s, 3H), 4.3 (s, 2H), 6.8 (s, 2H) Example 3 Metabolic stability of ambroxol analogs

[0150] The metabolic stability of ambroxol and its analogs was studied in the presence of human liver microsomes.

[0151] Study Design - Test compounds (ambroxol and its analog) were dissolved in dimethyl sulfoxide (DMSO) to prepare 1 mM stock solutions. The compounds were added to human liver microsome preparations (200 μL per incubation at a protein concentration of 1 mg / mL) to a final compound concentration of 1 μM. Samples were incubated at 37°C in the presence of a reduced nicotinamide adenine dinucleotide phosphate (NADPH) regeneration system (NADP, 1 mM, pH 7.4; glucose-6-phosphate, 5 mM, pH 7.4; glucose-6-phosphate dehydrogenase, 1 unit / mL). A positive control (midazolam, 1 μM, 0.1 mg / mL protein) was co-incubated to ensure that the microsome preparations performed as expected. Samples were acquired at t=0, 30, 60, and 120 minutes and quenched by the addition of acetonitrile.

[0152] Quenched samples were analyzed by liquid chromatography (LC / MS-MS) connected to tandem mass spectrometry detection. The concentration of the test compound at each time point was converted to the remaining percentage relative to the concentration of the compound at time t=0 (which acts as the 100% value). All data points were included in the data processing. The elimination rate constant (k) for each test compound was calculated. el , minutes -1 ) is a single exponential decay equation (A t =A0e -k el t , where A t is the remaining percentage at time t, A0 is 100%, and k el The in vitro half-life (t) was determined from the change in the rate of disappearance of the parent compound over time, based on the fitting of experimental data to the disappearance rate constant (where t is time). 1 / 2 ) to, k el From the formula: t 1 / 2 =ln(2) / k el The determination was made based on the following: Using the elimination rate constant, the in vitro intrinsic clearance CL was determined. int - microsomes (μL / min / mg protein) was also calculated.

[0153] Results - Estimated in vitro elimination half-lives (t) for compounds ZW-010 and ZW-011. 1 / 2 ) and inherent clearance (CL int - microsomes The values ​​are shown in Table 2, and here they are compared with the values ​​obtained for the parent compound, ambroxol. [Table 2]

[0154] As shown in Table 2, the substitution of two bromine substituents of ambroxol, such as ZW-010 (two fluorine substituents) and ZW-011 (two methoxy substituents), leads to improved metabolic stability, as the elimination half-lives of both compounds are more than double that of ambroxol. This improvement in stability is unexpected. The increased stability of the analogs may allow for higher exposure to the analogs when administered. In addition, the improved stability of the analogs may allow for a reduction in the dosing schedule compared to ambroxol alone. In summary, these improvements may lead to an improvement in the overall cost of the analogs by achieving sustained efficacy while reducing the required dose levels and dose frequencies compared to ambroxol. Example 4 Comparative pharmacokinetics (PK) of ambroxol and ZW-010

[0155] After acclimatizing the mice in the laboratory for a minimum of three days, female C57BL / 6 mice (6-8 weeks old, 16-30g, N=21 mice per group) were orally administered a single dose of ambroxol or ZW-010 as their hydrochloride salts via force feeding. Both compounds were administered at a dose of 10 mg / kg (free base equivalent) as a clear solution of 1 mg / mL in 5:95 v / v dimethyl sulfoxide (DMSO) / saline. The volume administered was calculated for each animal based on their recent body weight. Animals were fasted overnight before drug administration and food and water were returned 4 hours after dose administration. Mice (N=3 mice per group) were sacrificed at 0.15, 0.25, 1, 2, 4, 8, and 24 hours after administration. Blood (in a K2EDTA collection tube) and brain were collected from each animal at each time point. Samples for quantitative analysis (internal standard: verapamil) by HPLC / MS-MS (C18 column, gradient elution; standard curves from 1 to 3,000 ng / mL; electrospray, positive ionization mode, multiple reaction monitoring-MRM-mode) were prepared using plasma (isolated from blood samples by centrifugation) and brain homogenate (obtained by homogenization using 50% methanol / water in a 1:2 w / v ratio). The concentrations (in units ng / mL for plasma and ng / g for brain) were calculated, and the mean and standard deviations for plasma and brain are plotted as a function of time after injection in Figures 2A (ambroxol) and 2B (ZW-010).

[0156] As shown in Figures 2A and 2B, both compounds are cerebral penetrating agents with a brain / plasma ratio greater than 1 at all time points. Although both compounds were administered at the same dose (10 mg / kg), exposure to ZW-010 in both plasma and brain was also observed to be significantly greater than exposure to ambroxol. These results demonstrate that the halogenated analog of ambroxol improves exposure in both brain and periphery due to its improved stability in the body. These properties make the novel compounds ideal for treating disorders involving both neurological and peripheral systems as described herein. [Table 3]

[0157] As shown in Table 3, brain and plasma exposure to ZW-010 is higher than that to ambroxol. Both compounds are cerebral penetrating agents, and the brain / plasma ratio is greater than 1.0, with a larger ratio for ambroxol (11.0) than for ZW-010 (3.6). However, the absolute brain exposure (in ng.h / g) to ZW-010 is higher than that to ambroxol. It should be noted that the molar exposure concentration to ZW-010 is even higher (AUClast = 42.0 for ZW-010 and 12.3 μM.h for ambroxol, respectively). In addition, the elimination half-life (t1 / 2) of ZW-010 was approximately twice as long as that of ambroxol.

[0158] Therefore, the improved metabolic stability of ZW-010 compared to ambroxol (see Example 3) leads to improved pharmacokinetics of ZW-010 compared to ambroxol, with significantly higher exposure to brain tissue when administered at the same dose and with a significantly longer elimination half-life. These results suggest the potential of the compound of the present invention to achieve efficacy at significantly lower doses and with a better dosing schedule (once or twice daily) than required for ambroxol, thereby limiting the risk of off-target side effects and potentially improving the commercial cost of manufacturing the compound.

[0159] Overall, these data suggest that this novel ambroxol analog will require less frequent and lower doses than the parent compound. This is significant considering that clinical trials of high-dose ambroxol regimens tended to have dropouts due to poor patient adherence. See Istaiti, M. et al., "High-Dose Ambroxol Therapy in Type 1 Gaucher Disease Focusing on Patients with Poor Response to Enzyme Replacement Therapy or Substrate Reduction Therapy," Int. J. Mol. Sci. 2023, 24, 6732 and NCT02941822. Example 5 Bioactivity of the test compound

[0160] In Figure 3, human neurally induced pluripotent stem cells (iPSCs) were obtained from the New York Stem Cell Foundation. Cultured cells were differentiated and maintained in a proprietary medium from Stem Cell Technology. Compounds including ambroxol, difluoro-substituted ambroxol (compound 1; also referred to herein as "ZW-010"), and dimethoxy-substituted ambroxol (compound 5; also referred to herein as "ZW-011") were lysed in DMSO. Cells were treated for 14 days. Lysosomes were stained with "Lysotracker Green" (Invitrogen) according to the manufacturer's instructions. Cells were fixed, and a 3D image stack was obtained using a Leica SP8 laser scanning confocal microscope. Lysosome size was quantified using IMARIS 7.7 software (Bitplane). Human neurons were obtained from (Kwart D, Gregg A, Scheckel C, Murphy EA, Paquet D, Duffield M, Fak J, Olsen O, Darnell RB, Tessier-Lavigne M: A Large Panel of Isogenic APP and PSEN1 Mutant Human iPSC Neurons Reveals Shared Endosomal Abnormalities Mediated by APP beta-CTFs, Not Abeta. Neuron 2019, 104:256-270 e255. 31416668).

[0161] In Figure 4, N2a neuroblastoma cells were obtained from the American Tissue Type Collection (ATCC). The cells were cultured in 6-well plates at a density of 100,000 cells / well. The cells were placed in MEM supplemented with 10% fetal bovine serum (FBS) for 24 hours, and then differentiated in MEM by serum starvation for 24 hours. The cells were then treated in MEM supplemented with 10% FBS with DMSO, ambroxol, or difluoro-substituted ambroxol (compound 1; also referred to herein as "ZW-010") and dimethoxy-substituted ambroxol (compound 5; also referred to herein as "ZW-011"). The cells were left in the treated medium for 24 hours, and then RNA was extracted with TRIzol-chloroform. cDNA was synthesized using the iScript cDNA synthesis kit (Bio-Rad), and RT-PCR was performed using the SsoAdvanced Universal SYBR Green Supermix kit (Bio-Rad). Difluoro and dimethoxy compounds increased the transcription of lysosome-related LAMP1, cathepsin B, GBA1, and autophagy-related LC3 and TFEB to a greater extent than ambroxol.

[0162] In Figure 5, human iPSC-derived neurons (as shown in Figure 3) were cultured in two 6-well plates at a density of 150,000 cells / well. On day 6 of culture, one 6-well plate was treated with an equal volume of DMSO (vehicle control), and the other 6-well plate was treated with 10 mM [micromolar concentration] of ambroxol or difluoro-substituted ambroxol (compound 1; also referred to herein as "ZW-010") and dimethoxy-substituted ambroxol (compound 5; also referred to herein as "ZW-011") during the first 50 / 50 medium change. Cells were treated for 3 days, and cells were extracted with TRIzol-chloroform. cDNA was synthesized using the iScript cDNA synthesis kit (Bio-Rad), and RT-PCR was performed using the SsoAdvanced Universal SYBR Green Supermix kit (Bio-Rad). Difluoro and dimethoxy compounds increased the transcription of lysosome-related LAMP1, cathepsin D, cathepsin B, GBA1, and autophagy-related LC3 and TFEB to a greater extent than ambroxol.

[0163] Figures 6A-B show that difluoro-substituted ambroxol (compound 1; also referred to herein as "ZW-010") induces TFEB migration to the nucleus. Human iPSC-derived neurons were treated with difluoro-substituted ambroxol for 14 days. Cells were fixed and immunostained for endogenous TFEB. Nuclei were identified by DAPI staining. Cells were imaged as a 3D stack using a Leica SP8 confocal microscope. TFEB signals appeared as points throughout the cytoplasm and nucleus (Figure 6A). Nuclear TFEB points were quantified using IMARIS 7.7 software (Bitplane) (Figure 6B).

[0164] Figures 7A-B show the differentiation of human neurons derived from iPSCs bearing the Swedish mutation, followed by maintenance using culture media from Stem Cell Technologies. Cells were treated with DMSO (solvent), ambroxol, or difluoro-substituted ambroxol (compound 1; also referred to herein as "ZW-010") for 5 days. The culture media were collected and analyzed for A-beta 40 (Figure 7A) and 42 (Figure 7B) by ELISA (Invitrogen) according to the manufacturer's instructions for use. Both ambroxol and its derivatives were effective in reducing the secretion of beta-amyloid species. Example 6 The compound may be shown to improve lifespan.

[0165] Two-month-old weaned male BDF1 mice, raised from pups and housed in cages of 1-4 animals, were freely fed Teklad 7013 NIH-31 rodent chow diet and water. These mice were divided into two nearly equal groups of 22-25 animals each. One group continued to be fed the Teklad chow diet as before (control), while the other group was switched to a Teklad chow diet formulated with one of compounds 1-6 at 300 mg per kg of chow diet. This formulation was designed to deliver 50 mg / kg / day (based on body mass) to each mouse in the treatment group, based on the average free chow diet consumption of adult male BDF1 mice. Mice were maintained on this diet until they died naturally or reached 16 months of age (at which point the experiment ended). Throughout the experiment, all mice were periodically removed from their cages and handled in the process of being subjected to various sensorimotor, cognitive, and / or behavioral tests.

[0166] Figure 8 provides animal data for experiments using ambroxol as described in the preceding paragraph. This shows that ambroxol can increase lifespan in a mouse model. Group 1 represents control animals that were not administered ambroxol, and Group 2 represents animals that were administered 50 mg / kg of ambroxol as a daily chow supplement, starting at 2 months of age. The data suggest that not only did ambroxol increase lifespan, but that the surviving animals in Group 2 appeared to be at least as healthy as the animals in Group 1 on average when sacrificed, indicating an extension of healthy lifespan in parallel with lifespan. Compounds 1 to 6 are expected to achieve similar results. Example 7 Research using the compound of the present invention, which shows activity in improving tremors in a mouse model of Parkinson's disease (PD).

[0167] The compounds of the present invention can be investigated for their activity in improving tremors in a mouse animal model of Parkinson's disease, as follows.

[0168] Select 6-OHDA mice exhibiting resting tremor (i.e., mice injected with 6-hydroxydopamine in the striatum). Three groups of mice, each containing at least 10 mice, will be administered either a "high dose" (Group 1: 150 mg / kg / day of the compound of the present invention), a "low dose" (Group 2: 50 mg / kg / day of the compound of the present invention), or no administration of the compound of the present invention (control group). Tremors in the mice will be monitored daily using electromyography or force plate-based measurements (Bekar L et al., Nat Med 14:75-80, 2008) to evaluate the effect of the compound of the present invention on resting tremor. Example 8 Research using the compound of the present invention that shows efficacy against aggregated Aβ in a mouse model of Alzheimer's disease (AD).

[0169] The compounds of the present invention can be investigated for their activity against amyloid-beta peptide (Aβ) aggregates and deposits in a mouse animal model of Alzheimer's disease as follows. Figure 11 shows the results of testing ambroxol (Figure 11B) versus control (Figure 11A) in the following mouse model. Using this same model, the analogs described herein can also be tested, and it is predicted that if administered at the same dose for a significantly longer period in other models, they will show similar, or otherwise better, results due to improved pharmacokinetics and significantly higher exposure in brain tissue.

[0170] Select an AD mouse model that shows significantly elevated Aβ production (e.g., mice containing the "Swedish mutation" in human amyloid precursor protein (APP)). Using two groups of mice, namely a "treatment" group (introduced 2400 mg / kg of the compound of the present invention into animal feed) and a control group (without the compound of the present invention), after an appropriate treatment period (e.g., 2 months of treatment in animals old enough to show substantial AD-related pathology), the generalization of aggregated Aβ and the relative development of fibrous deposits can be analyzed by comparing, for example, silver staining or Aβ immunohistochemistry with Congo red or thioflavin-S histology (Jankowsky J et al., Mol Neurodegen 12:89, 2017). Example 9 Research using the compound of the present invention that exhibits activity in improving cognitive function

[0171] The compounds of the present invention can be investigated for their activity in improving cognitive function in mouse animal models.

[0172] Cognitive sensitivity tests were administered over seven months to three groups of mice, each containing at least 13 mice. The three groups were a "high-dose" group (150 mg / kg / day of the compound of the present invention), a "low-dose" group (50 mg / kg / day of the compound of the present invention), and a control group (not administered the compound of the present invention).

[0173] The results of such experiments, but using ambroxol instead of the compound of the present invention, are shown in Figure 9. A dose-related improvement in cognitive acuity was observed, with the “high-dose” group showing the best cognitive acuity results, the “low-dose” group showing results between the control and “high-dose” groups, and the control group showing baseline cognitive acuity results. The dosage results were substantially lower than the doses previously observed to effectively promote GC-ase chaperoning activity in mice (e.g., Migdalska-Richards et al., Ann Neurol 80: 766-775, 2016). Example 10 Research using the compound of the present invention that induces macroautophagy

[0174] The compounds of the present invention can be investigated for their ability to induce macroautophagy in mouse cells in culture medium as follows.

[0175] Mouse fibroblasts (NIH3T3) were obtained from the American Type Culture Collection (ATCC). The cells were maintained at 37°C with 5% CO2 in Dulbecco's Modified Eagle Medium (DMEM) (Sigma, St. Louis, MO) in the presence of 10% neonatal bovine serum (NCS), 50 μg / ml penicillin, and 50 μg / ml streptomycin. The cells, cultured in 96-well glass-bottom plates, were treated for the indicated time, fixed, and then imaged using a high-content microscope (Operetta, PerkinElmer). For example, images of nine different regions per well could be captured, yielding an average of 2,500–3,000 cells. Nuclei and dots were identified using the manufacturer's software. The number of particles / dots per cell could be quantified in the cytoplasmic region using a "particle identifier" function after thresholding in unsaturated images. In all cases, the focal plane thickness is set to 0.17 μm, and sections with the largest nuclear diameter are selected for quantification. The value may be presented as the number of points per cell section representing 10-20% of the total points per cell under the acquisition conditions. Macroautophagy activity in intact cells is measured during transduction with a lentivirus carrying the mCherry-GFP-LC3 tandem construct (Kimura S et al., Autophagy 3(5):452-460, 2007). Cells are cultured in a glass-bottom 96-well plate, and fluorescence is read in both channels. Points positive for both fluorophores correspond to autophagosomes, while points positive only for the red fluorophore correspond to autolysosomes. The autophagy flux is determined as the conversion from autophagosomes (yellow) to autolysosomes (red points only).

[0176] Figure 10 shows the results of a comparable experiment using ambroxol. Ambroxol appears to inhibit nutrient sensing, aside from its pharmacological activity as a GC-ase chaperone. By interfering with nutrient sensing, ambroxol can induce cellular responses in animals appropriate to entering nutritional restriction or fasting conditions, thus directing the organism into a “catabolic signaling” mode characterized by lysosomal biosynthesis and autophagy induction, potentially leading to improvements in lifespan, healthy lifespan, and / or cognitive sharpness (see, e.g., Efeyan et al., Nature 517:302-310, 2015) (the whole is incorporated herein by reference). Therefore, ambroxol and analogues of ambroxol and related compounds can systemically inhibit nutrient sensing, resulting in the entire organism entering a catabolic signaling mode.

[0177] The human equivalent doses (HEDs) calculated from the "low" and "high" mouse doses in this study are approximately 4 mg / kg / day and 12 mg / kg / day, respectively, which are approximately 250 mg / day and 750 mg / day for a human with an average weight of 62.5 kg. Therefore, in a preferred embodiment, the doses would be approximately 50 mg / day, 75 mg / day, 100 mg / day, 150 mg / day, 200 mg / day, 250 mg / day, 300 mg / day, 350 mg / day, 400 mg / day, 450 mg / day, 500 mg / day, 550 mg / day, 600 mg / day, 650 mg / day, 700 mg / day, 750 mg / day, 800mg / day, 850mg / day, 900mg / day, 950mg / day, 1000mg / day, 1050mg / day, 1100mg / day, 1150mg / day, 1200mg / day, or 50-150mg / day, 50-200mg / day, 50-250mg / day, 250-500mg / day, 250-1000mg / day, 1500-2 Long-term administration of compounds of formula I (or pharmaceutically acceptable salts, solvates, or prodrugs thereof) administered at doses of 000 mg / day, 1000–1500 mg / day, or between 1000–2000 mg / day, or less than 1000 mg / day, or approximately 1 mg / kg / day, 2 mg / kg / day, 3 mg / kg / day, 4 mg / kg / day, 5 mg / kg / day, 6 mg / kg / day, 7 mg / kg / day, 8 mg / kg / day, 9 mg / kg / day, 10 mg / kg / day, 11 mg / kg / day, 12 mg / kg / day, and / or between 4–12 mg / kg / day may be effective in improving healthy lifespan, life expectancy, and / or intellectual sharpness. Example 11 Formulation of the compound of formula I into a high-drug-load liquid oral pharmaceutical composition and its properties

[0178] Preparation of Granules - A suitable granule of any one of compounds 1 to 6 (as representative examples of the compounds of the present invention) may be prepared as follows: A finely ground compound in powder form is placed in a rotor granulator (GXR-35 rotor granulator, Freund-Vector Corporation), and a binder solution of hydroxypropyl cellulose (HPC-Klucel LF) is sprayed onto the powder to form a granular core. Additional compound powder is then co-sprayed with the binder solution to grow spheres. As layers of compound are added, the particles become more spherical. The resulting spheres may contain 97% by weight of compound I and 3% by weight of HPC. A particle size of about 350 microns (×50) and a density of about 0.7 g / ml can be achieved.

[0179] Application of water-soluble seal coating to granules - Compound A granules can be seal-coated using a bottom-spray fluidized bed coater equipped with a Wolster column to produce a smooth and uniform substrate. The batch size may be approximately 750 grams. A suitable seal coating contains 9.1% by weight hypromellose (HPMC), 0.9% by weight triethyl citrate (TEC), and 90% by weight water. Seal-coating the granules can result in a 2% weight increase.

[0180] Application of enteric coating to granules - Sealed compound granules may be enterically coated using a bottom-spray fluid bed coater equipped with a Wolster column. The batch size may be approximately 750 grams. A suitable enteric coating contains 58.0% by weight of Eudragit L30D55, 0.9% by weight of triethyl citrate (TEC), 8.7% by weight of Plasacryl T20, and 32.5% by weight of water. Enteric coating of sealed granules can result in a 35% weight increase.

[0181] Solubility Properties - Enteric-coated granules can be tested for enteric solubility properties. Table 4 shows the solubility parameters of equivalent granules containing ambroxol hydrochloride. [Table 4]

[0182] All patents, patent applications, and publications listed herein are thus incorporated herein by reference in their entirety.

[0183] While the present invention has been disclosed with reference to specific embodiments, it will be apparent that other embodiments and variations of the present invention can be devised by those skilled in the art without departing from the true spirit and scope of the invention. The appended claims are intended to be construed as encompassing all such embodiments and equivalent variations.

Claims

1. Formula I: 【Transformation 5】 A compound or a pharmaceutically acceptable salt, solvate, or prodrug thereof, wherein the formula is: ... indicates that the associated R or X group may be bonded to any available carbon atom on the phenyl ring. R ａ H, hydroxyl (OH), lower alkyl (e.g., CH ３ and CH ２ CH ３ C etc. １～３ Alkyl alcohols, and lower alcohols (e.g., CH ２ C such as OH １～３ Selected from alcohol, R ｂ is selected from H and lower alkyl (e.g., CH ３ and CH ２ CH ３ such as C １～３ alkyl), R ｃ and R ｄ These are, independently, H and a lower alkyl (e.g., CH). ３ and CH ２ CH ３ C etc. １～３ Selected from alkyl, or R ｃ and R ｄ These are, respectively, R ｃ and R ｄ It is part of a 4, 5, 6, or 7-membered ring structure that connects them (for example, -R ｃ -N-R ｄ - (CH2) ｎ (where n is an integer selected from 1, 2, 3, and 4) R １ From R １４ Each of them is independently selected from H and D, X １ and X ２ These are independently F, Cl, Br, I, and lower alkyl (e.g., CH ３ and CH ２ CH ３ C etc. １～３ Alkyl), lower alkoxy (e.g., OCH ３ and OCH ２ CH ３ C etc. １～３ Alkoxy), lower alkylamines (e.g., NR ｂ CH ３ and NR ｂ CH ２ CH ３ C etc. １～３ Alkylamines), lower acyls (e.g., C(O)CH ３ and C(O)CH ２ CH ３ C etc. ２～４ Acyl), Nitro (NO ２ ), nitrile (CN), sulfoxide (SO-R), sulfonate (SO2-R), and sulfate (O-SO ２ -O-R) is selected, however X １ and X ２ It is not possible for both to be Br. A compound or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

2. Equation Ia below: 【Transformation 6】 The compound according to claim 1, having the following characteristics.

3. X １ and X ２ The compound according to any one of claims 1 to 2, wherein all of them are F.

4. X １ and X ２ However, all of them are OCH ３ The compound according to any one of claims 1 to 2.

5. X １ and X ２ The compound according to any one of claims 1 to 2, wherein all of them are Cl.

6. R ａ OH is R ｂ The compound according to any one of claims 1 to 5, wherein is H.

7. R ａ H is R ｂ ga CH ３ The compound according to any one of claims 1 to 5. 【Request Item 8】 【Chemistry 7】 A compound according to any one of claims 1 to 2, selected from the above.

9. The aforementioned compound 【Transformation 8】 A compound according to any one of claims 1 to 2, selected from the above.

10. A pharmaceutical composition comprising a compound according to any one of claims 1 to 9 (or a pharmaceutically acceptable salt, solvate, or prodrug thereof), in combination with a pharmaceutically acceptable carrier as necessary.

11. The pharmaceutical composition according to claim 9, wherein the composition is a liquid oral pharmaceutical composition.

12. A pharmaceutical composition according to claim 10, comprising a high-load compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) as described in any one of claims 1 to 9, wherein the composition comprises (i) the compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) and (ii) at least one pharmaceutically acceptable excipient, wherein the compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) is in the form of a granule having a granule core comprising about 60 to about 97 weight percent of the active pharmaceutical ingredient and about 3 to about 40 weight percent of the excipient, wherein the weight percentage is based on the total weight of the granule core.

13. A pharmaceutical composition according to claim 10, comprising a high-load compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) as described in any one of claims 1 to 9, wherein the composition comprises (i) the compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof), (ii) at least one pharmaceutically acceptable excipient, and (iii) a diluent, wherein the compound of formula I (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) is in the form of granules having a granule core comprising about 60 to about 97 weight percent of the active pharmaceutical ingredient and about 3 to about 40 weight percent of the excipient, wherein the weight percent is based on the total weight of the granule core, wherein the granule core is coated with (iv) a water-soluble seal coating in an amount that provides a weight increase of about 0.5 to about 5 percent, and (v) an enteric coating in an amount that provides a weight increase of about 0.5 to about 50 percent.

14. A method for preventing and / or treating a disease or medical condition selected from the group consisting of respiratory diseases and conditions, lysosomal storage disorders (LSDs), and neurological diseases and conditions, wherein the method comprises the step of administering to the subject an effective amount of a compound according to any one of claims 1 to 9 (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) or a pharmaceutical composition according to any one of claims 10 to 13.

15. The method according to claim 14, wherein the disease or medical condition to be prevented and / or treated is a bronchopulmonary disease, or Gaucher disease, Pompe disease, or Fabry disease, or Parkinson's disease, Lewy body dementia, Alzheimer's disease (AD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS) (i.e., Lou Gehrig's disease), frontotemporal dementia (FTD), Pick's disease, or Gaucher disease.

16. A method comprising the step of administering to a subject an effective amount of a compound according to any one of claims 1 to 9 (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) or a pharmaceutical composition according to any one of claims 10 to 13, wherein the method is for the purpose of extending the life expectancy of the subject, or treating, inhibiting, or reducing aging in the subject, or treating, inhibiting, or reducing age-related symptoms or age-related diseases in the subject, or increasing the healthy lifespan, life expectancy, and / or intellectual sharpness of the subject.

17. A method for preventing and / or treating, reducing the symptoms of, and / or slowing the progression of, Alzheimer's disease (AD) or other diseases associated with pathogenic protein misfolding, aggregation, and deposition (including Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS) (i.e., Lou Gehrig's disease), Pick's disease, Gaucher's disease, and frontotemporal degeneration (FTD)) in a subject, wherein the method comprises the step of administering to the subject an effective amount of ambroxol (or related compounds such as ambroxol hydrochloride and bromhexine) or a compound according to any one of claims 1 to 9 (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) in combination with one or more suitable anti-beta-amyloid antibodies or fragments thereof.

18. A method for preventing, reducing the symptoms of, and / or slowing the progression of, Alzheimer's disease (AD) or other diseases associated with pathogenic protein misfolding, aggregation, and deposition (including Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS) (i.e., Lou Gehrig's disease), Pick's disease, Gaucher's disease, and frontotemporal degeneration (FTD)), comprising the step of administering to a subject an effective amount of ambroxol (or related compounds such as ambroxol hydrochloride and bromhexine) or a compound according to any one of claims 1 to 9 (or a pharmaceutically acceptable salt, solvate, or prodrug thereof).

19. The method according to claim 17 or 18, wherein the subjects are selected by assaying for biomarkers indicating patients at risk of or in the early stages of AD or other amyloid diseases associated with pathogenic protein misfolding, aggregation, and deposition (including Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS) (i.e., Lou Gehrig's disease), Pick's disease, Gaucher's disease, and frontotemporal degeneration (FTD) diseases).

20. The method according to claim 19, wherein the subjects are selected by assaying for phosphorylated tau protein (p-tau) that indicates a patient at risk of AD or a patient in the early stages of AD.

21. The method according to claim 17 or 18, wherein the subject is selected by genotyping of at least one gene or locus that points to a patient at risk of AD or other diseases associated with pathogenic protein misfolding, aggregation, and deposition (including Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS) (i.e., Lou Gehrig's disease), Pick's disease, Gaucher's disease, and frontotemporal degeneration (FTD) diseases).

22. The method according to claim 21, wherein the subject is selected by genotyping the ApoE gene or by assaying for p-tau217, p-tau181, p-tau231, p-tau235, and / or N3pG.

23. The method according to claim 22, wherein the subject is selected by genotyping the ε4 allele of the ApoE gene, or by assaying for p-tau217, p-tau181, p-tau231, p-tau235, and / or N3pG.

24. The ambroxol (or related compound) or the compound described in any one of claims 1 to 9 (or a pharmaceutically acceptable salt, solvate, or prodrug thereof) is administered to the subject. (i) A dosage or amount that provides a peak concentration in the serum of the subject greater than 1 μM, for example, 2 to 50 μM, 2 to 25 μM, or 10 to 20 μM. (ii) A dosage that provides a peak concentration in the brain tissue of the subject greater than 3 μM, for example, 5-50 μM, 5-25 μM, or 10-20 μM, or (iii) Dosage within the range of approximately 250-1000 mg / day, 750-1000 mg / day, 1000-2000 mg / day, 1000-1500 mg / day, or 1500-2000 mg / day. The medication is administered in a daily dose selected from the following: The method according to any one of claims 17 to 23.

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