Oral delivery of active compounds for treating glycogen storage diseases - Patents.com

Abstract

The present disclosure relates to formulations for oral delivery of at least one active agent, a compound according to Formula (I) and / or (II), for the treatment of glycogen storage diseases, neurodegenerative disorders, and autophagy-related conditions, wherein the formulation is a nanostructured formulation to increase the bioavailability of the active agent after administration.

Description

[Technical Field]

[0001] The present disclosure relates to formulations for oral delivery of at least one active agent for the treatment of glycogen storage diseases, neurodegenerative disorders and / or autophagy-related conditions. [Background technology]

[0002] References believed to be relevant background to the subject matter disclosed herein are listed below: -PCT Patent Application Publication No. WO2018 / 154578 -PCT Patent Application No. PCT / IL2022 / 050187 -Kapetanovic et al., “Effects of oral dosing paradigms (gavage versus diet) on pharmacokinetics and pharmacodynamics”, 2006, Chem Biol Interact, 164, 68-75 -Kleyweg et al., “Interobserver Agreement In The Assessment Of Muscle Strength And Functional Abilities In Guillain- Barre Syndrome”, 1991, Muscle & Nerve, 14, 1103-1109 Acknowledgment of the above references herein should not be inferred to mean that they are in any way relevant to the patentability of the subject matter disclosed herein.

[0003] background Glycogen is a branched polysaccharide in which glucose units are linked by α1-4 glycosidic bonds to form linear chains and further linked by α1-6 glycosidic bonds to form branching junctions.

[0004] Glycogen is stored primarily in the liver and muscle, but may also be found at lower levels in the kidney, heart, and brain. In the liver, it provides a source of energy during fasting periods, while in muscle, glycogen serves as an immediate reserve source of available glucose.

[0005] Glycogen synthesis begins with the autoglycosylation of the oligosaccharide primer of glycogenin. Glycogen elongation involves glycogen synthase, which catalyzes the formation of the α1-4 glycosidic bond, and glycogen branching enzyme (GBE), which catalyzes the formation of the α1-6 glycosidic bond and glycogen. Glycogen degradation occurs via two distinct pathways: one in the cytosol (termed "glycogenolysis") and one in the lysosome, termed "glycogen autophagy" or "glycophagy." Glycogen breakdown via glycogenolysis involves glycogen phosphorylase (GP) and glycogen debranching enzyme (GDE), while the glycophagy mechanism is mediated by acid α-glucosidase (GAA).

[0006] The importance of glycogen metabolism is not only emphasized in the context of energy conservation, but is also associated with several congenital disorders caused by abnormal function of enzymes that control glycogen synthesis, degradation, and regulation. These disorders are most often inherited in an autosomal recessive manner and are collectively referred to as glycogen storage diseases (GSDs). Today, there are 16 known subtypes of GSD in the medical community, including Lafora disease and Danon disease, all of which manifest with a variety of clinical symptoms.

[0007] Recently, several compounds have been designed to treat GSD and are described in PCT Patent Publication WO2018 / 154578. These compounds have shown promising activity in the treatment of GSD. Summary of the Invention

[0008] The present disclosure provides a formulation for oral delivery of active compounds for treating GSD, as well as for treating neurodegenerative disorders or conditions associated with lysosomal storage or autophagy misregulation.The formulations disclosed herein are designed to allow for increased loading of active compounds, while maintaining high tolerability and improved bioavailability.

[0009] The disclosed formulations are formulated to stabilize the active compound in a water-free nanostructured formulation while allowing complete dilution in aqueous liquids, so that upon oral administration, the formulation disperses homogeneously in the aqueous phase (e.g., gastric fluid), forming dispersed nanostructures within which the active compound is trapped and stabilized. Such entrapment allows the active compound to be stabilized in the formulation, and upon administration, the active compound is released from the nanostructures, ensuring the delivery of high effective doses of the active compound over time. The inventors surprisingly discovered that the use of a combination of solvent(s), co-surfactant(s), and hydrophilic surfactant(s) (forming a substantially hydrophilic delivery system) enables the formation of nanostructures in the water-free formulation, as well as the in situ formation of nanostructures after administration, while stabilizing a high load of the lipophilic active compound disclosed herein.

[0010] According to one of its aspects, the present disclosure provides a pharmaceutical formulation for oral delivery of a compound of formula (I) or a pharmaceutically acceptable salt, isomer or tautomer thereof, [ka] [ka] represents a single or double bond, n and m are integers, each independently 1, 2, or 3; R and R 1 are each independently hydrogen or absent; R 2 , R 3 , R 4 , R 5 , R6 , R 7 , R 8 and R 9 are each hydrogen or each independently selected from alkyl, cycloalkyl, alkoxy, hydroxy, thiohydroxy, thioalkoxy, aryloxy, thioaryloxy, amino, nitro, halo, trihalomethyl, cyano, amido, carboxy, sulfonyl, sulfoxy, sulfinyl, and sulfonamido, each further substituted or unsubstituted; and One of X and Y is S and the other of X and Y is C; provided that if X is S, then R 9 does not exist and if Y is S, then R 5 provided that does not exist; The formulation comprises: a) the compound of formula (I) or a pharmaceutically acceptable salt, isomer, or tautomer thereof; b) at least one hydrophilic surfactant in a total amount ranging from about 10 wt% to 50 wt%; c) at least one solvent in a total amount of at least about 20 wt %; d) at least one co-surfactant; and e) at least one oil in an amount of 0 wt% to about 5 wt% of the formulation; The weight ratio of the at least one solvent to the at least one hydrophilic surfactant ranges from about 1.25:1 to about 1:3.

[0011] In some embodiments, the compound of Formula (I) is a compound of Formula (I') or a pharmaceutically acceptable salt, isomer, or tautomer thereof: [ka] [ka] represents a single or double bond, n and m are integers, each independently 1, 2, or 3; R and R 1 are each independently hydrogen or absent; and R 2 , R 3 , R 4 , R 5 , R 6 , R 7 and R 8 are each hydrogen, or R 2 , R 3 , R 4 , R 5 , R 6 , R 7 and R 8 are each independently selected from alkyl, cycloalkyl, alkoxy, hydroxy, thiohydroxy, thioalkoxy, aryloxy, thioaryloxy, amino, nitro, halo, trihalomethyl, cyano, amido, carboxy, sulfonyl, sulfoxy, sulfinyl, and sulfonamido, each of which is further substituted or unsubstituted.

[0012] The formulations of this disclosure are designed for oral delivery of the active compound, i.e., delivery of the active compound by swallowing, for systemic pharmacological effect. The formulations are typically in liquid form and can be administered as a liquid, gel, suspension, or encapsulated in a liquid gel or soft gel capsule.

[0013] Due to their unique formulation, the formulations of this disclosure can be stably loaded with the active compound at a concentration of at least 0.5 wt %, e.g., at least about 1 wt %, at least about 2 wt %, at least about 3 wt %, at least about 4 wt %, or even at least about 5 wt % of the formulation.

[0014] According to some embodiments, the formulation comprises up to about 10 wt% of said compound of formula (I) or a pharmaceutically acceptable salt, isomer, or tautomer thereof.

[0015] The inventors have found that the combination of hydrophilic surfactant(s), co-solvent(s) and solvent(s) allows for high loading of the active compound in the formulation and its stabilization for extended periods of time, while allowing for spontaneous formation of nanostructures both in water-free formulations and when mixed with aqueous fluids (e.g., gastric fluids after administration). In the formulations of this disclosure, the balance of material components allows for high loading and entrapment of lipophilic active agents in a primarily hydrophilic formulation for extended periods of time, allowing for long shelf-life and with minimal phase separation and / or sedimentation.

[0016] The formulations of this disclosure are typically in a concentrated form, typically a water-free concentrate, that is stably dilutable with aqueous media (i.e., there is no substantial increase in droplet size or phase separation upon dilution). The concentrated form is stable for extended periods of time, lacks a life-sustaining environment for microorganisms, and is readily dilutable in aqueous media as further described below.

[0017] Thus, the formulations disclosed herein are typically devoid of water and are designed to allow for the spontaneous formation of nanostructures in concentrated form (i.e., without the presence of water) and when mixed with an aqueous liquid (after or for the purpose of administration), as further detailed below.

[0018] In the disclosed formulations, a delicate balance between surfactant and co-surfactant / solvent confers physical stability to the formulation (lacking water) in the presence of high concentrations of pharmaceutically active compounds, as such balance has been found to promote solubilization of the active compounds. At the same time, the ratio between surfactant and co-surfactant / solvent allows for the formation of very small droplets, less than about 10 nm, thereby enabling the dilution capability of the formulation in aqueous liquids.

[0019] The term hydrophilic surfactant(s) refers to a surface-active agent having a hydrophilic head group and a lipophilic tail that can arrange into nanostructures in aqueous media. The inventors have found that the combination of hydrophilic surfactants with co-surfactants and solvents in a specific ratio range and total concentration can spontaneously form stable nanostructures, which stabilize the active compound in the formulation of a waterless concentrate and solubilize the active compound into nanostructures when mixed with an aqueous liquid.

[0020] According to some embodiments, the at least one hydrophilic surfactant is selected from ethoxylated fatty acids, ethoxylated castor oil and its hydrogenated derivatives, polysorbates, ethoxylated alkyl ethers, ethoxylated monoglycerides, polyglycerol esters, sucrose esters, and combinations thereof.

[0021] According to some embodiments, the formulation comprises at least one first hydrophilic surfactant selected from ethoxylated castor oil and its hydrogenated derivatives (e.g., polyoxyl 35 castor oil, polyoxyl 40 hydrogenated castor oil, polyoxyl 60 castor oil, polyoxyl 60 hydrogenated castor oil), and at least one second hydrophilic surfactant selected from polysorbates (polysorbate 20, polysorbate 60, polysorbate 80) and ethoxylated monoglycerides (caprylocaproyl polyoxyl-8 glyceride, lauryl polyoxyl 32 glyceride, stearoyl poloxyl 32 glyceride, etc.).

[0022] As noted above, the at least one hydrophilic surfactant is present in the formulation in an amount of about 10 wt% to about 50 wt%. According to some embodiments, the at least one hydrophilic surfactant is present in a total amount ranging from about 20 wt% to 50 wt%. According to other embodiments, the at least one hydrophilic surfactant is present in a total amount ranging from about 25 wt% to 50 wt%.

[0023] The formulation comprises at least one co-surfactant. The co-surfactant should be understood to include any hydrophilic, lipophilic, or amphiphilic agent different from the hydrophilic surfactant(s), which (together with the surfactant) contributes to reducing the interfacial tension between the oily phase and the aqueous phase to nearly zero (or even zero), allowing the formation of thermodynamically stable nanostructures. Thus, the combination of the surfactant and the co-surfactant allows the formulation to be stabilized both kinetically and thermodynamically.

[0024] According to some embodiments, the at least one co-surfactant is selected from polyethylene glycol 200, polyethylene glycol 400, polyethylene glycol 600, propylene glycol, phospholipids (e.g., phosphatidylcholine), diethylene glycol monoethyl ether, and combinations thereof.

[0025] According to some embodiments, the at least one co-surfactant is present in the formulation in a total amount ranging from about 8 wt% to about 45 wt%. According to some other embodiments, the at least one co-surfactant is present in the formulation in a total amount ranging from about 8 wt% to about 30 wt%, or even from about 8 wt% to 25 wt%.

[0026] According to some embodiments, the weight ratio between the total hydrophilic surfactants and the total co-surfactants in the formulation ranges from about 3:1 to about 1:3. According to some embodiments, the weight ratio between the total hydrophilic surfactants and the total co-surfactants in the formulation ranges from about 3:1, 2.9:1, 2.8:1, 2.7:1, 2.6:1, 2.5:1, 2.4:1, 2.3:1, 2.2:1, 2.1:1, 2.0:1, 1.9:1, 1.8:1, 1.7:1, 1.6:1, 1.5:1, 1.4:1, 1.5:1, 1.6:1, 1.8 ... .3:1, 1.2:1, 1.1:1, 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2, 1:2.1, 1:2.2, 1:2.3, 1:2.4, 1:2.5, 1:2.6, 1:2.7, 1:2.8, 1:2.9, or approximately 1:3.

[0027] According to some embodiments, the weight ratio between the total hydrophilic surfactants and the total co-surfactants in the formulation ranges from about 2.5:1 to about 1:1.5. According to further embodiments, the weight ratio between the total hydrophilic surfactants and the total co-surfactants in the formulation ranges from about 2:1 to about 1:1.

[0028] The formulation also contains a relatively large amount, typically at least 20 wt %, of at least one solvent, which is an organic solvent, typically polar, that is at least partially water-miscible and suitable for aiding in the solubilization of the active compound in the formulation and into the nanostructures.

[0029] According to some embodiments, the formulation comprises the at least one solvent in a concentration ranging from about 20 wt% to about 45 wt%. According to some embodiments, the formulation comprises the at least one solvent in a concentration ranging from about 20 wt% to about 35 wt%.

[0030] According to some embodiments, the at least one solvent is selected from ethanol, methanol, n-propanol, benzyl alcohol, and combinations thereof.

[0031] According to some embodiments, the total amount of solvent and co-surfactant in the formulation is at least about 45 wt%. According to some embodiments, the total amount of solvent and co-surfactant in the formulation is at least about 50 wt%. According to other embodiments, the total amount of solvent and co-surfactant in the formulation is at least about 52 wt%. According to some other embodiments, the total amount of solvent and co-surfactant in the formulation is at least 55 wt%.

[0032] According to some embodiments, the weight ratio between the total solvent and the total co-surfactant ranges from about 3:1 to 1:2. According to some embodiments, the weight ratio of the total amount of solvent to the total amount of co-surfactant ranges from about 2:1 to about 1:1.5.

[0033] According to some embodiments, the weight ratio of the total amount of solvent to the total amount of co-surfactant is about 3:1, 2.8:1, 2.6:1, 2.4:1, 2.2:1, 2:1, 1.8:1, 1.6:1, 1.4:1, 1.2:1, 1:1, 1:1.2, 1:1.4, 1:1.6, 1:1.8, or about 1:2.

[0034] As noted above, the weight ratio of the total amount of solvent to the total amount of hydrophilic surfactant ranges from about 1.25:1 to about 1:3. According to some embodiments, the weight ratio of the total amount of solvent to the total amount of hydrophilic surfactant ranges from about 1:1 to about 1:2.

[0035] According to some embodiments, the weight ratio of the total amount of solvent to the total amount of hydrophilic surfactant is about 1.25:1, 1.2:1, 1.15:1, 1.1:1, 1.05:1, 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, 1:2, 1:2.1, 1:2.2, 1:2.3, 1:2.4, 1:2.5, 1:2.6, 1:2.7, 1:2.8, 1:2.9, or about 1:3.

[0036] The formulation comprises 0 wt% to about 5 wt% oil. According to some embodiments, at least one oil is present in the formulation at a concentration of 4 wt% or less. According to some embodiments, the formulation is devoid of oil.

[0037] The term oil refers to a lipophilic agent that is immiscible in water and can form distinct domains when introduced into an aqueous liquid. In some embodiments, at least one oil is selected from short-chain triglycerides and medium-chain triglycerides.

[0038] In some embodiments, the formulations may further include various additives approved for pharmaceutical use, such as pH adjusters and buffers, neutralizing agents, emollients, humectants, preservatives, antioxidants, taste masking agents, taste modifiers, sweeteners, flavor additives, and any other suitable non-active pharmaceutical additives.

[0039] The formulations of the present disclosure are designed as pharmaceutical formulations for oral delivery of a compound of formula (I) or a pharmaceutically acceptable salt, isomer, or tautomer thereof.

[0040] According to some embodiments, n and m in formula (I) are 1.

[0041] According to some embodiments, R in (I) 2 , R 7 and R 8 The formula is methyl.

[0042] According to some embodiments, the compound of formula (I) is at least one compound of formula (IA) or (IB): [ka]

[0043] According to another embodiment, the formulation comprises two or more compounds of formula (I).

[0044] As used herein, alkyl, alkenyl, and alkynyl carbon chains, unless otherwise specified, contain 1 to 20 carbons and can be straight or branched. Alkenyl carbon chains of 2 to 20 carbons, in certain embodiments, contain 1 to 8 double bonds, and alkenyl carbon chains of 2 to 16 carbons, in certain embodiments, contain 1 to 5 double bonds. Alkynyl carbon chains of 2 to 20 carbons, in certain embodiments, contain 1 to 8 triple bonds, and alkynyl carbon chains of 2 to 16 carbons, in certain embodiments, contain 1 to 5 triple bonds. Exemplary alkyl, alkenyl, and alkynyl groups herein include, but are not limited to, methyl, ethyl, propyl, isopropyl, isobutyl, n-butyl, sec-butyl, tert-butyl, isohexyl, allyl (propenyl), and propargyl (propynyl).

[0045] C 1-6Alkyl should be understood to include any straight or branched chain alkyl moiety having 1, 2, 3, 4, 5 or 6 carbon atoms.

[0046] Cycloalkyl, in certain embodiments, refers to a saturated monocyclic or polycyclic ring system of 3 to 10 carbon atoms, and in other embodiments, 3 to 6 carbon atoms; cycloalkenyl and cycloalkynyl refer to a monocyclic or polycyclic ring system containing at least one double bond and at least one triple bond, respectively. Cycloalkenyl and cycloalkynyl groups may, in certain embodiments, contain 3 to 10 carbon atoms, cycloalkenyl groups in further embodiments contain 4 to 7 carbon atoms, and cycloalkynyl groups in further embodiments contain 8 to 10 carbon atoms. The ring systems of cycloalkyl, cycloalkenyl, and cycloalkynyl groups may be composed of one ring or two or more rings, which may be joined together in a fused, bridged, or spiro-bonded fashion.

[0047] Heterocyclyl, in one embodiment, refers to a 3- to 10-membered mono- or polycyclic non-aromatic ring system in which one or more of the atoms in the ring system is a heteroatom, i.e., an element other than carbon, such as, but not limited to, nitrogen, oxygen, or sulfur.

[0048] Alkoxy refers to -O-alkyl or -O-cycloalkyl, as defined herein; thioalkoxy refers to -S-alkyl or -S-cycloalkyl, as defined herein.

[0049] Aryl denotes an aromatic monocyclic or polycyclic carbon group containing 5 to 19 carbon atoms, ie having a conjugated π electron system.

[0050] Heteroaryl, in certain embodiments, refers to an about 5 to about 19-membered monocyclic or polycyclic aromatic ring system in which one or more of the atoms in the ring system is a heteroatom, i.e., an element other than carbon, such as, but not limited to, nitrogen, oxygen, or sulfur.

[0051] Aryloxy refers to -O-aryl or -O-heteroaryl, as defined herein; thioaryloxy refers to -S-aryl or -S-heteroaryl, as defined herein.

[0052] Hydroxy refers to the -OH group.

[0053] Thiohydroxy (or thiol) refers to the group -SH.

[0054] Amino refers to a primary, secondary, or tertiary amine (-NR'R", where R' and R" are independently hydrogen, alkyl, cycloalkyl, alkenyl, aryl, heteroaryl, or heterocyclyl as defined herein), and the point of attachment is through the nitrogen atom, which is substituted with a C1-C6 straight or branched alkyl. In the case of tertiary amines, the substituents may be the same or different.

[0055] Nitro refers to the -NO2 group.

[0056] Halo (or halogen or halide) refers to F, Cl, Br, or I. Haloalkyl refers to an alkyl group in which one or more of the hydrogen atoms has been replaced with a halogen. Such groups include, but are not limited to, trihalomethyl.

[0057] Cyano refers to the group -C≡N.

[0058] Amide refers to the divalent group -C(O)NH2.

[0059] Carboxy refers to the group -C(O)-OR', where R' is hydrogen, alkyl, cycloalkyl, alkenyl, aryl, heteroaryl, or heterocyclyl, as defined herein.

[0060] Sulfonyl refers to the group -S(O)2-R', where R' is hydrogen, alkyl, cycloalkyl, alkenyl, aryl, heteroaryl, or heterocyclyl, as defined herein.

[0061] Sulfinyl refers to the group -S(O)-R', where R' is hydrogen, alkyl, cycloalkyl, alkenyl, aryl, heteroaryl, or heterocyclyl, as defined herein.

[0062] Sulfonamide refers to the group -S(O)2-NR'R" where R' and R" are independently hydrogen, alkyl, cycloalkyl, alkenyl, aryl, heteroaryl, or heterocyclyl as defined herein.

[0063] The term "pharmaceutically acceptable salt(s)" as used herein refers to salts of compounds of this disclosure that are safe in mammals, effective for pharmaceutical use, and possess the desired biological activity. Pharmaceutically acceptable salts include salts of acidic or basic groups present in compounds of this disclosure. Pharmaceutically acceptable acid addition salts include, but are not limited to, hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, superphosphate, isonicotinic acid, acetate, lactate, salicylate, citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)). Certain compounds of the invention can form pharmaceutically acceptable salts with various amino acids. Suitable base salts include, but are not limited to, aluminum, calcium, lithium, magnesium, potassium, sodium, zinc, and diethanolamine salts.

[0064] The compounds described herein may contain one or more chiral atoms, or may otherwise exist as isomers, such as two enantiomers or two or more diastereomers. Thus, the compounds may contain mixtures of isomers as well as purified isomers or enantiomerically enriched mixtures. Furthermore, the compounds may contain mixtures of diastereomers as well as purified stereoisomers or diastereomerically enriched mixtures. It should also be noted that the compounds may form tautomers in isolation or any mixture thereof.

[0065] According to another aspect of the disclosure, there is provided a pharmaceutical formulation for oral delivery of a compound of formula (II) or a pharmaceutically acceptable salt, isomer, or tautomer thereof, [ka] The formulation comprises: a) the compound of formula (II) or a pharmaceutically acceptable salt, isomer or tautomer thereof; b) at least one hydrophilic surfactant in a total amount ranging from about 10 wt% to 70 wt%; c) at least one solvent in a total amount of at least about 15 wt %; d) at least one co-surfactant, and e) at least one oil in an amount of 0 wt% to about 5 wt% of the formulation; The weight ratio of the at least one solvent to the at least one hydrophilic surfactant ranges from about 1:1 to about 1:7.

[0066] According to some embodiments, the formulation may comprise a mixture of at least one compound of formula (I) and a compound of formula (II).

[0067] According to some embodiments, the formulation comprises up to about 10 wt% of said compound of formula (II) or a pharmaceutically acceptable salt, isomer, or tautomer thereof.

[0068] According to some embodiments, the at least one hydrophilic surfactant is selected from ethoxylated fatty acids, ethoxylated castor oil and its hydrogenated derivatives, polysorbates, ethoxylated alkyl ethers, ethoxylated monoglycerides, polyglycerol esters and sucrose esters, and combinations thereof. According to some embodiments, the formulation comprises at least one first hydrophilic surfactant selected from ethoxylated castor oil and its hydrogenated derivatives (e.g., polyoxyl 35 castor oil, polyoxyl 40 hydrogenated castor oil, polyoxyl 60 castor oil, polyoxyl 60 hydrogenated castor oil), and at least one second hydrophilic surfactant selected from polysorbates (polysorbate 20, polysorbate 60, polysorbate 80) and ethoxylated monoglycerides (caprylocaproyl polyoxyl-8 glyceride, lauryl polyoxyl 32 glyceride, stearoyl poloxyl 32 glyceride, etc.).

[0069] According to some embodiments, the at least one hydrophilic surfactant is present in a total amount ranging from about 20 wt% to 70 wt% of the formulation of Compound (II). According to other embodiments, the at least one hydrophilic surfactant is present in a total amount ranging from about 30 wt% to 70 wt%. According to some embodiments, the formulation of the compound of Formula (II) comprises said at least one hydrophilic surfactant in a total amount ranging from about 20 wt% to 50 wt%.

[0070] According to some embodiments, the at least one co-surfactant is selected from polyethylene glycol 200, polyethylene glycol 400, polyethylene glycol 600, propylene glycol, phosphatidylcholine, diethylene glycol monoethyl ether, and combinations thereof.

[0071] According to some embodiments, the at least one co-surfactant is present in the formulation of the compound of Formula (II) in a total amount ranging from about 8 wt% to about 45 wt%. According to other embodiments, the at least one co-surfactant is present in the formulation of the compound of Formula (II) in a total amount ranging from about 8 wt% to about 35 wt%.

[0072] According to some embodiments, the weight ratio between the total hydrophilic surfactants and the total co-surfactants in the formulation of a compound of Formula (II) ranges from about 7:1 to about 1:3. According to some embodiments, the weight ratio between the total hydrophilic surfactants and the total co-surfactants in the formulation ranges from about 7:1 to about 1:1, e.g., about 7:1, 6.5:1, 6:1, 5.5:1, 5:1, 4.5:1, 4:1, 3.5:1, 3:1, 2.5:1, 2:1, 1.5:1, or 1:1.

[0073] Formulations of the compound of Formula (II) also contain a relatively large amount, typically at least 15 wt %, of at least one solvent. According to some embodiments, formulations of the compound of Formula (II) contain the at least one solvent in a concentration ranging from about 15 wt % to about 45 wt %. According to some embodiments, formulations of the compound of Formula (II) contain the at least one solvent in a concentration ranging from about 15 wt % to about 35 wt %.

[0074] According to some embodiments, the at least one solvent is selected from ethanol, methanol, n-propanol, benzyl alcohol, and combinations thereof.

[0075] According to other embodiments, the weight ratio of the at least one solvent to the at least one hydrophilic surfactant in the formulation of the compound of Formula (II) ranges from about 1.1:1 to about 1:5. In some embodiments, the weight ratio of the at least one solvent to the at least one hydrophilic surfactant ranges from about 1.1:1 to about 1:3.

[0076] According to some embodiments, the total amount of solvent and co-surfactant in the formulation of the compound of Formula (II) is at least about 25 wt %.

[0077] Formulations of compounds of formula (II) contain from 0 wt% to about 5 wt% oil, preferably up to 2 wt% oil, and even more preferably are devoid of oil.

[0078] In some embodiments, formulations of the compounds of Formula (I), Formula (I') and / or Formula (II) may further include various additives approved for pharmaceutical use, such as pH adjusters and buffers, neutralizing agents, emollients, humectants, preservatives, antioxidants, taste masking agents, taste modifiers, sweeteners, flavor additives, and any other suitable non-active pharmaceutical additives.

[0079] The formulations of the compounds of Formula (I), Formula (I') and / or Formula (II) of the present disclosure are designed as pharmaceutical formulations for oral delivery of the compounds of Formula (I), Formula (I') and / or Formula (II) or pharmaceutically acceptable salts, isomers or tautomers thereof.

[0080] As discussed above, the formulations of the compounds of Formula (I), Formula (I'), and / or Formula (II) of this disclosure are designed to be stable (thermodynamically and kinetically) for extended periods of time. The formulations of this disclosure are in concentrated form (i.e., devoid of water) and form nanostructures upon mixing with an aqueous liquid. The nanostructures stabilize and entrap the active compound, allowing for its containment within the formulation prior to dilution with the aqueous liquid and its release from the nanostructures after administration (i.e., after dilution).

[0081] A careful balance between the hydrophilic surfactant(s), co-surfactant(s), and solvent(s) allows for the spontaneous formation of nanostructures in which the active compound is solubilized and stabilized. The combination of hydrophilic surfactant(s), co-surfactant(s), and solvent(s) promotes complete coverage of the interface between the nanostructures and the aqueous diluent at high water dilutions of the formulation. In other words, the combination of hydrophilic surfactant, co-surfactant, and solvent changes the effective critical packing parameter (ECPP) of the interface, facilitating control of the hydrophilic / hydrophobic nature of the surfactant with the amount of water, thereby increasing the stability of the nanostructures.

[0082] The formulations of this disclosure can be administered as is, i.e., in concentrated form, and can be easily diluted in situ by gastric fluids after administration. Alternatively, the formulations can be administered in diluted form by diluting the formulation with one or more aqueous diluents prior to administration.

[0083] Thus, according to another of its aspects, the present disclosure provides a preparation for oral delivery of a compound of formula (I) or a pharmaceutically acceptable salt, isomer or tautomer thereof, the preparation comprising nanodroplets of a formulation comprising said compound of formula (I) or a pharmaceutically acceptable salt, isomer or tautomer thereof as disclosed herein dispersed in a continuous phase comprising at least one aqueous diluent.

[0084] A further aspect provides a preparation for oral delivery of a compound of formula (I') or a pharmaceutically acceptable salt, isomer or tautomer thereof, the preparation comprising nanodroplets of a formulation comprising said compound of formula (I') disclosed herein or a pharmaceutically acceptable salt, isomer or tautomer thereof dispersed in a continuous phase comprising at least one aqueous diluent.

[0085] A further aspect provides a preparation for oral delivery of a compound of formula (II) or a pharmaceutically acceptable salt, isomer or tautomer thereof, the preparation comprising nanodroplets of a formulation comprising said compound of formula (II) or a pharmaceutically acceptable salt, isomer or tautomer thereof dispersed in a continuous phase comprising at least one aqueous diluent.

[0086] It should generally be noted that unless specifically stated otherwise, the term formulation is used to denote a composition that does not contain water (i.e., a concentrated form), and the term preparation is meant to denote a diluted form of a formulation.

[0087] Nanodroplets (or nanostructures) are droplets composed of formulations that trap and stabilize pharmaceutically active compounds. Nanostructures are typically in the form of droplets with an average diameter of up to 50 nm (nanometers), where hydrophilic surfactants and cosurfactants form the interface between the continuous solvent phase (in the case of a water-free concentrated form) or the continuous aqueous phase and the oil core. Without wishing to be bound by theory, the active compound is located at the interface, so that at least some of the active compound is physically trapped between the head groups of the cosurfactants and stabilized within the nanostructure.

[0088] According to some embodiments, the nanodroplets have an average droplet size in the range of about 5 nm to 50 nm.

[0089] The term mean size refers to the arithmetic mean of the measured diameters of the droplets. If the droplets are not spherical, the mean size calculation is based on an equivalent sphere for the largest dimension of the particle.

[0090] According to some embodiments, the nanodroplets are substantially monodisperse. Formulations and preparations are typically transparent (or substantially transparent) due to the size of their monodispersed submicron nanostructures, and their transparency is maintained over time. This allows for easy detection of changes in the stability of the formulation and / or preparation (as phase separation, bioactive precipitation, and / or coalescence of the droplets causes detectable turbidity).

[0091] According to some embodiments, the at least one aqueous diluent is selected from water, water for injection, saline, dextrose solution, and buffer solution.

[0092] Another aspect provides a formulation or preparation as disclosed herein for use in treating glycogen storage disease (GSD).

[0093] According to a further aspect, there is provided a method of treating glycogen storage disease (GSD) comprising administering to a patient in need thereof an effective amount of a formulation or preparation disclosed herein.

[0094] According to some embodiments, GSD is associated with glycogen-branching enzyme deficiency, which refers to a disease or disorder characterized by the deposition, accumulation, or aggregation of polyglucosan bodies in muscle, nerve, and / or other tissues of the body.

[0095] According to some other embodiments, the GSD is GSD Type 0, GSD Type I, GSD Type II, GSD Type III, GSD Type IV, GSD Type V, GSD Type VI, GSD Type VII, GSD Type VIII, GSD Type IX, GSD Type X, GSD Type XI, GSD Type XII, GSD Type XIII, GSD Type XIV, or GSD Type XV.

[0096] According to other embodiments, the GSD is adult polyglucosan body disorder (APBD), Andersen's disease, Forbes' disease, or Danon's disease.

[0097] A further aspect of this disclosure provides a formulation or preparation as disclosed herein for use in treating a disease or condition associated with lysosomal storage.

[0098] According to another further aspect, the disclosure provides a method of treating a disease or condition associated with lysosomal storage comprising administering to a patient in need thereof an effective amount of a formulation or preparation disclosed herein.

[0099] Lysosomal storage disorders (LSDs) are a group of genetic diseases characterized by lysosomal dysfunction and neurodegeneration. These disorders are typically caused by a single gene defect in a specific enzyme that is primarily required for the normal degradation of glycosaminoglycans (GAGs). Such defects result in the inability of cells to excrete carbohydrate residues, leading to their accumulation in intracellular lysosomes and thus disruption of normal cell function. Exemplary lysosomal storage disorders include sphingolipidoses, ceramidases (e.g., Farber disease, Krabbe disease), galactosialidoses, gangliosidoses, e.g., α-galactosidases (e.g., Fabry disease (α-galactosidase A), Schindler disease (α-galactosidase B)), β-galactosidases (e.g., GM1 gangliosidosis, GM2 gangliosidosis, Sandhoff disease, Tay-Sachs disease), glucocerebrosidoses (e.g., Gaucher disease (types I, II, III)), sphingomyelinases (e.g., lysosomal acid lipase deficiency, Niemann-Pick disease), sulfatidoses (e.g., metachromatic leukodystrophy), and erythropoietin-related disorders. Mucopolysaccharidoses (e.g., Type I (MPSI (Hurler syndrome, Scheie syndrome, Hurler-Scheie syndrome), Type II (Hunter syndrome), Type III (Sanfilippo syndrome), Type IV (Morquio syndrome), Type VI (Maroteaux-Lamy syndrome), Type VII (Sly syndrome), Type IX (hyaluronidase deficiency)), mucolipidoses (e.g., Type I (sialidosis), Type II (I-cell disease), Type III (pseudo-Hurler polydystrophy / phosphotransferase deficiency), Type IV (mucolipidin 1 deficiency)), lipidoses (e.g., Niemann-Pick disease), neuronal ceroid lipofuscinoses (e.g., Type 1 Santavuori-Haltia disease / infantile NCL (CLN1 PPT1)), Type 2 Jansky-Bielschowsky disease / late-onset childhood NCL (CLN2 / LINCL))TPP1), Batten-Spielmeyer-Vogt disease type 3 / juvenile NCL (CLN3), Kufs disease type 4 / adult NCL (CLN4), Finnish variant / late-onset childhood type 5 (CLN5), late-onset childhood variant type 6 (CLN6), CLN7 type 7, Northern epilepsy type 8 (CLN8), Turkish late-onset childhood type 8 (CLN8), German / Serbian late-onset childhood type 9, congenital cathepsin D deficiency type 10 (CTSD), Wolman disease, oligosaccharidoses (e.g., alpha-mannosidosis, beta-mannosidosis, aspartylglucosaminuria, fucosidosis), lysosomal transport diseases (e.g., cystinosis, pyknodysostosis, Salla disease / sialic acid storage disease, infantile free sialic acid storage disease), Pompe disease type II, Danon disease type lib), cholesteryl ester storage disease, etc.

[0100] In some embodiments, the disease or condition associated with lysosomal storage is selected from Gaucher disease, Fabry disease, Tay-Sachs disease, mucopolysaccharidosis (MPS) disorders, aspartylglucosaminuria, GMI-gangliosidosis, Krabbe (globoid cell leukodystrophy or galactosylceramide lipidosis), metachromatic leukodystrophy, Sandhoff disease, mucolipidosis type II (I-cell disease), mucolipidosis type IIIA (pseudo-Hurler polydystrophy), Niemann-Pick disease types C2 and C1, Danon disease, free sialic acid storage disorders, mucolipidosis type IV, multiple sulfatase deficiency (MSD), metabolic disorders, obesity, type II diabetes, and insulin resistance.

[0101] According to yet another aspect, there is provided a formulation or preparation disclosed herein for use in treating a disease or condition associated with autophagy-misregulation.

[0102] According to another further aspect, there is provided a method of treating a disease or condition associated with autophagy-misregulation, comprising administering to a patient in need thereof an effective amount of a formulation or preparation disclosed herein.

[0103] Autophagy is a catabolic process involving the degradation of cell components, such as long-lived proteins, protein aggregates, organelles, cell membranes, organelle membranes, and other cellular components. The mechanism of autophagy can involve: (i) the formation of a membrane around a target region of the cell, separating the contents from the rest of the cytoplasm; (ii) the fusion of the resulting vesicle with lysosomes and subsequent degradation of the vesicle contents. Diseases associated with autophagy misregulation can include diseases caused by misfolded protein aggregates, such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, Huntington's disease, spinocerebellar ataxia, oculopharyngeal muscular dystrophy, prion disease, fatal familial insomnia, α1-antitrypsin deficiency, dentatorubral-pallidoluysian atrophy, frontotemporal dementia, progressive supranuclear palsy, X-linked spinal muscular atrophy, and neuronal intranuclear hyaline inclusion disease. Also included are diseases or disorders such as cancer, cardiovascular, neurodegenerative, metabolic, pulmonary, renal, infectious, musculoskeletal, and ocular disorders, in which the induction of autophagy can contribute to delaying the onset and slowing, stopping, or reversing the progression of one or more symptoms associated with the disease or disorder. Diseases associated with autophagy misregulation also include cancer, for example, any cancer in which the induction of autophagy inhibits cell growth and division, reduces mutagenesis, removes mitochondria and other organelles damaged by reactive oxygen species, or kills developing tumor cells. The term is further intended to include psychiatric diseases or disorders, such as any psychiatric disease or disorder in which the induction of autophagy can contribute to delaying the onset and slowing, stopping, or reversing the progression of one or more symptoms associated with the psychiatric disease or disorder. In one embodiment, the psychiatric disease or disorder is selected from schizophrenia and bipolar disorder.

[0104] According to some embodiments, the disease or condition associated with autophagy-misregulation is selected from Alzheimer's disease and cancer associated with reduced autophagy activity.

[0105] According to some other embodiments, the disease or condition is a neurodegenerative disease, e.g., selected from Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), multiple sclerosis, Huntington's disease, spinocerebellar ataxia, oculopharyngeal muscular dystrophy, multiple system atrophy with Lewy bodies, and prion diseases.

[0106] As is known, the effective amount for the purposes herein can be determined by such considerations known in the art. The amount must be effective to achieve the desired therapeutic effect, depending, inter alia, on the type and severity of the disease to be treated and the treatment regime. The effective amount is typically determined in a well-designed clinical trial (dose-ranging study), and those skilled in the art will know how to properly conduct such a study to determine the effective amount. As is generally known, the effective amount depends on various factors, such as various pharmacological parameters, such as half-life in the body, any undesirable side effects, factors such as age and sex, etc.

[0107] The term treatment or any linguistic variation thereof, as used herein, refers to the administration of a therapeutic amount of a formulation or preparation of the present disclosure that is effective to ameliorate undesirable symptoms associated with the disease, prevent the onset of such symptoms before they occur, slow the progression of the disease, slow the worsening of symptoms, enhance the onset of remission periods, slow the irreversible damage caused in the progressive chronic phase of the disease, delay the onset of said progressive phase, reduce the severity or cure the disease, improve survival or more rapid recovery, or prevent the disease from occurring, or a combination of two or more of the above.

[0108] The term subject is meant to denote a mammal, human or non-human.

[0109] As used herein, the term about is meant to encompass a deviation of ±10% from the specifically stated value of a parameter, e.g., temperature, concentration, etc.

[0110] Unless otherwise specified, all concentrations disclosed herein are provided as a weight percentage, wt%, from the weight of the formulation or preparation, as the case may be.

[0111] Whenever a numerical range is given herein, it is meant to include any recited number (fractional or integer) within the recited range. The phrases ranging / ranges between a first designated number and a second designated number and ranging / ranges from a first designated number to a second designated number are used interchangeably herein and are meant to include the first and second designated numbers and all fractional and integer numbers therebetween.

[0112] Unless otherwise required by context, the word comprise and variations such as "comprises" and "comprising" will be understood to imply the inclusion of a stated integer or step or group of integers or steps, but not the exclusion of any integer or step or group of integers and steps.

[0113] The term at least one as applied to any component of a formulation should be read to encompass one, two, three, four or more different occurrences of said component in a formulation or preparation.

[0114] It will be appreciated that certain features of the invention that are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, for simplicity, various features of the invention that are described in the context of a single embodiment may also be provided, separately or in any suitable subcombination, as suitable in any other described embodiment of the invention. Certain features described in the context of various embodiments should not be considered essential features of those embodiments, unless the embodiment is inoperable without those elements.

[0115] For a better understanding of the subject matter disclosed herein and to illustrate how it may be carried out in practice, embodiments will now be described, by way of non-limiting example only, with reference to the accompanying drawings, in which: [Brief explanation of the drawings]

[0116] [Figure 1] 1A-1F show photographs of exemplary formulations according to several embodiments of the present disclosure. [Figure 2] Figures 2A-2C show droplet size (Z-average) (●) and PDI (■) measurements as a function of compound GHF-201 concentration in the formulations: 8CS-RG (Figure 2A), LDS-C (Figure 2B), and LDS-600 (Figure 2C). [Figure 3] Figures 3A-3C show viscosity as a function of the concentration of compound GHF-201 in the formulations: 8CS-RG (Figure 3A), LDS-C (Figure 3B), and LDS-600 (Figure 3C). [Figure 4] Figures 4A-4B. Refractive index RI (Figure 4A) and normalized RI (Figure 4B) viscosity as a function of the concentration of compound GHF-201 in the formulations: 8CS-RG (●), LDS-C (■), and LDS-600 (▲). [Figure 5] Figures 5A-5L. Compound GHF-201: 8CS-RG vehicle (Figure 5A), 8CS-RG 2.5% GHF-201 (Figure 5B), 8CS-RG 5.0% GHF-201 (Figure 5C), 8CS-RG 7.5% GHF-201 (Figure 5D), LDS-C vehicle (Figure 5E), LDS-C 2.5% GHF-201 (Figure 5F), LDS-C 5.0% GHF-201 (Figure 5G), LDS-C 7.5% GHF-201 (Figure 5H), LDS-600 vehicle (Figure 5I), LDS-600 2.5% GHF-201 (Figure 5J), LDS-600 5.0% GHF-201 (Figure 5K), LDS-600 LUMiFuge test results for an exemplary formulation loaded with 7.5% GHF-201 (FIG. 5L). [Figure 6]Figures 6A-6C. Photographs of formulation LDS-C, 7.15% compound GHF-201, from physical stability studies: t=0 (Figure 6A), after 12 months at 25°C (Figure 6B), and after 12 months at 40°C (Figure 6C). [Figure 7] Figures 7A-7C show DLS analysis results for formulation LDS-C, 7.15% compound GHF-201, in physical stability studies: t=0 (Figure 7A), after 12 months at 25°C (Figure 7B), and after 12 months at 40°C (Figure 7C). [Figure 8] 8A-8B. Tissue penetration study results in APBD model mice for compound GHF-201: unformulated (FIG. 8A) and formulated in LDS-C formulation, 7.5 wt% (FIG. 8B). [Figure 9] Figures 9A-9C. Pharmacokinetic profiles of compound GHF-201 administered orally to APBD patient 1 in formulation LDS-C (Figure 9A) and IV at 250 mg / kg to n=3 APBD model mice (Figure 9B). The active ingredient doses on days 1, 2, and 3 were 170 mg, 255 mg, and 340 mg, respectively. Values ​​in boxes A are expanded in Figure 9C. [Figure 10] Figures 10A-10C. Pharmacokinetic profiles of compound GHF-201 administered orally to APBD patient 2 in formulation LDS-C (Figure 10A) and IV at 250 mg / kg to n=3 APBD model mice (Figure 10B). The active ingredient doses on days 1, 2, and 3 were 170 mg, 255 mg, and 340 mg, respectively. Values ​​are expanded in Figure 10C. [Figure 11] 11A-11B. Area under the curve (AUC) correlated with daily administered dose for patients 1 and 2, respectively. [Figure 12] 12A-12C. Muscle strength grade (before treatment and after 10 months of treatment) for patient 1 (FIG. 12A) and plasma neurofilament light chains for patient 1 (FIG. 12B) and patient 2 (FIG. 12C) treated with a formulation of compound GHF-201. DETAILED DESCRIPTION OF THE INVENTION

[0117] Exemplary Formulations A blank formulation was prepared according to Table 1 by weighing all the ingredients and mixing them at 40-60°C.

[0118] The compound of formula (IA), herein designated as Compound GHF-201 (or alternatively as Compound A), was selected as an exemplary compound: [ka] The compound GHF-201 was then solubilized in the formulations at various concentrations (2.5, 5.0, 7.5 wt%, weight percentage of the weight of the empty formulation). [Table 1] [Table 2]

[0119] Physical characterization Formulations (such as those in Table 1 above) are devoid of water and are completely dilutable in aqueous liquids. Upon dilution, the formulations form nanostructures with an average droplet size of approximately 20 nm.

[0120] Selected formulations from Table 1 were analyzed to evaluate their physical properties. Exemplary visualizations of the appearance of some of the formulations can be seen in Figures 1A-1F: the formulations were in undiluted form (Figures 1A-1C) and diluted (90% water content) (Figures 1D-1E), and loaded with no compound GHF-201 ("vehicle"), 2.5 wt%, 5 wt%, and 7.5 wt% of compound GHF-201. As can be seen, all formulations exhibit high transparency, homogeneity, and thermodynamic stability in both undiluted and diluted forms.

[0121] The hydrodynamic radius of the droplets was measured at room temperature by dynamic light scattering (DLS) using a Nano-ZS Zetasizer (Malvern, UK) with water as the dispersant; the results are shown in Table 2 and Figures 2A-2C. [Table 3]

[0122] As seen in Table 2 and Figures 2A-2C, increasing the concentration and solubilization of compound GHF-201 in the formulations caused droplet shrinkage and a slight increase in PDI. For the empty formulation (vehicle), the average droplet size varied from 13 to 14 nm, while for the formulations loaded with compound GHF-201, the droplet sizes were approximately 10-11 nm and 9-10 nm for 2.5% and 5-7.5% compound GHF-201, respectively. The measured PDI values ​​were approximately 0.03-0.06 for the vehicle and up to 0.11 for 7.5% compound GHF-201. Overall, the PDI values ​​were low for both the vehicle and loaded formulations, indicating a relatively monodisperse system.

[0123] Without wishing to be bound by theory, it is hypothesized that the solubilization of compound GHF-201 results in stronger interactions between the cosurfactant and the solvent at the droplet interface. Increasing the concentration of compound GHF-201 results in the formation of more droplets of smaller size, leading to a gradual decrease in the average droplet size and an increase in polydispersity.

[0124] To further understand the interactions between compound GHF-201 and excipients in the formulation, viscosity and NMR measurements were performed.

[0125] Viscosity measurements were performed using an RS6000 rheometer (Thermo Scientific) equipped with a C60 / 1 TiL-L12007 cone, operated in rotational mode within the shear rate range of 0.01000-100.01 / s for 6 minutes. All measurements were performed at 25±1°C. Under these conditions, the formulations exhibited Newtonian behavior, and thus, they can be characterized by their viscosity. The viscosity of the systems was calculated based on a linear fit of rotational shear (τ) versus shear rate (γ). The slope of the linear fit represents the viscosity. Table 3 and Figures 3A-3C summarize the viscosities measured for the 8CS-RG, LDS-C, and LDS-600 system preconcentrates. [Table 4]

[0126] As shown in Figures 3A-3C, increasing the concentration of compound GHF-201 increased the viscosity of the formulation. Without wishing to be bound by theory, the presence of compound GHF-201 in all investigated formulations resulted in stronger interactions between the API and excipients, primarily cosurfactants, which contributed to the formulation's greater resistance to applied shear. This indicates that the introduction of compound GHF-201 into a formulation increases its stability and also its resistance to dynamic changes.

[0127] The pH and refractive index were measured for three exemplary formulations. The results are summarized in Table 4. For pH measurements, samples were diluted 10 times with purified water; the results in Table 4 are normalized to the pH of water. [Table 5]

[0128] As shown in Table 4, pH was not affected by the solubilization of compound GHF-201. Regarding refractive index (RI), it exhibited linear behavior as a function of compound GHF-201 concentration, with a similar slope of approximately 0.001 (1%) for all systems. RI has been shown to be affected by the viscosity of the system, so as viscosity increases, RI also increases. In this case, viscosity also increases with increasing compound GHF-201 content, so this trend in RI as a function of compound GHF-201 concentration is due to viscosity. Interestingly, when RI is normalized by viscosity, the resulting quotient decreases as a function of compound GHF-201 concentration (see Figures 4A-4B). Without wishing to be bound by theory, these results may be due to the nanostructured interface, which becomes more "saturated" with compound GHF-201, thus affecting the refraction of light.

[0129] The mobility of the components of the system was also measured by PGSE-NMR. The diffusion coefficients (×10) of several excipients and excipient groups were -11 ) are shown in Tables 5-1 to 5-3. Measurements were performed under a gradient / pulse program: δ = 10.0 ms, Δ = 27.0, gmax = 44.79. The D values ​​for surfactants and oils are the averages calculated for the two surfactants in each formulation and MCT. [Table 6] [Table 7] [Table 8]

[0130] Tables 5-1, 5-2, and 5-3 summarize the diffusion coefficients (D) of surfactants and oil (MCT), as well as the individual diffusion coefficients of PG, EtOH, GHF-201, and water. For LDS-600, the diffusion coefficient of PEG400 was also calculated. Overall, a clear trend of decreased diffusivity of all tested components as a function of GHF-201 concentration in concentrated form was observed in all tested formulations, suggesting that gradual loading of GHF-201 consistently strengthens interactions between all components. Surprisingly, when samples were diluted, the diffusivity of GHF-201 decreased, while surfactant mobility increased in all tested systems. This indicates that GHF-201 plays an important role as a structure builder in the system, helping to stabilize nanodroplets in the presence of water. This also supports the observations from DLS measurements, where the droplets progressively shrink as the GHF-201 content increases. The mobility of the solvent and cosurfactant in the dilute system, on the other hand, remained constant, suggesting that the role of these components is to stabilize the concentrate rather than the dilute system.

[0131] long-term physical stability To determine the long-term stability of the formulation, rapid measurements were performed using the LUMiSizer® analytical centrifuge. Results are shown in Figures 5A-5L. LUMiSizer® analysis can predict the shelf life of a formulation at its original concentration, even in the case of slow destabilization processes such as sedimentation, flocculation, coalescence, and fractionation. During a LUMiSizer® measurement, collimated light illuminates the entire sample cell within the centrifugal field; transmitted light is detected by sensors positioned linearly along the entire length of the sample cell. Local changes in particle or droplet size are detected by changes in light transmittance over time. Results are shown in graphs plotting the percentage of transmitted light (% transmittance) as a function of local position (mm), revealing the corresponding transmittance profile over time.

[0132] Changes in permeability indicate the stability of the formulation - if the permeability profile remains constant, the samples are considered physically stable and their shelf life can be extrapolated based on the measurement conditions.

[0133] As shown in Figures 5A-5L, all transmittance profiles overlapped, indicating that no changes in transmittance were observed, and all formulations were physically stable and, based on this analysis, are expected to be stable for at least 2 years under storage conditions.

[0134] Physical stability was also evaluated for formulation LDS-C with 7.15 wt% of compound GHF-201 for 12 months at 25°C and 40°C. Stability test results are provided in Tables 6-1 and 6-2. Sample appearances are shown in Figures 6A-6C, and DLS analysis results are shown in Figures 7A-7C. [Table 9] [Table 10]

[0135] The test results demonstrate that the formulation is physically stable at storage temperatures for at least 12 months with no evidence of significant changes in physical properties.

[0136] tissue penetration Figures 8A-8B show tissue penetration study results for compound GHF-201 in unformulated form (Figure 8A) and formulated in the LDS-C formulation (Figure 8B) in APBD model mice. For pharmacokinetic analysis, 100 μl of serum and brain, kidney, hind leg quadriceps, heart, liver, and spleen tissues were collected, homogenized, and extracted with acetonitrile according to established guidelines (Kapetanovic et al., 2006). A calibration curve was generated using 0, 1, 10, 100, and 1,000 ng / ml GHF-201 in a 1 mg / ml solution of 4-tert-butyl-2-(4H-1,2,4-triazol-4-yl)phenol (ChemBridge) as the internal standard (IS). Tissue samples were then dissolved in 1 mg / ml IS solution, and 0-1,000 ng / ml GHF-201 was added to generate a calibration curve from which tissue levels of GHF-201 were determined. Samples were analyzed by LC-MS / MS on a Sciex Triple Quad™ 5500 mass spectrometer.

[0137] Gbeys / ys mice were subcutaneously injected with 250 mg / kg of GHF-201 and sacrificed at 30, 60, 90, and 210 minutes after injection. The indicated tissues were removed, and 100 μL of serum was collected. The graph shows the mean (±SEM) GHF201 levels in different tissues as determined by LC-MS / MS. Results were obtained from n=3 mice at each time point.

[0138] The distribution and kinetic parameters of GHF-201 were evaluated in different tissues. Comparison of Figures 8A and 8B shows that the bioavailability of orally administered GHF-201 in a formulation according to the present disclosure was increased by at least 3-fold in all tissues compared to subcutaneous injection of GHF-201 in solution.

[0139] Pharmacokinetic studies Adult polyglucosan body disease (APBD) is a rare neurodegenerative disorder that most frequently affects adults of Ashkenazi Jewish origin. APBD is characterized by partial deficiency of glycogen branching enzyme 1 (GBE1) activity, most commonly caused by a mutational substitution of tyrosine 329 with a serine residue (Y329S). This loss-of-function mutation in GBE1 leads to the formation of abnormally structured glycogen aggregates called "polyglucosan bodies" (PBs, amylopectin-like polysaccharides with fewer branching points). Due to their solution and aggregation, PBs cannot be digested by the glycogen-digesting enzyme, glycogen phosphorylase. Accumulated aggregates often cause liver failure and death in childhood (Andresen's disease; glycogen storage disease type IV; GSDIV). Milder mutations in GBE1, such as p.Y329S, result in smaller PBs that do not disrupt hepatocytes and most other cell types, but simply accumulate on the sides of cells. However, in neurons and glial cells, these PBs clog the cramped confines of axons over time, leading to the debilitating and fatal progressive axonopathy disease APBD, which is often misdiagnosed as amyotrophic lateral sclerosis or multiple sclerosis. PB aggregates in APBD can cause a variety of phenotypic changes, including neurogenic bladder, partial limb motor dysfunction, lower body sensory dysfunction, and, in some cases, cognitive impairment. The advanced stages of the disease are characterized by difficulty walking, balance problems, and progressive weakness, and may even lead to death. Currently, there is no standard treatment for this condition.

[0140] The compound GHF-201 was found to be able to reduce polyglucosan in skin fibroblasts derived from APBD patients. Strict absorption, distribution, metabolism, excretion, and toxicity (ADMET) criteria were met in silico, and compound GHF-201 was found to be safe in mice (14-day study). Pharmacokinetically, when subcutaneously injected in mice, compound GHF-201 had high retention and persistence in the liver (>3 hours), intermediate levels and persistence in the brain and heart (1 hour), and negligible distribution to muscle. This pharmacokinetic profile was consistent with the histopathological effects of compound GHF-201 on individual tissues, with the greatest reduction in polyglucosan in the liver, moderate reductions in the brain and heart, and no effect in muscle. Continuous treatment of APBD mice with compound GHF-201 for 2 months before predicted disease onset significantly increased survival and improved motor and reflex parameters. The ameliorative effect on stretch reflexes is particularly important because the patient correlate is pyramidal tetraparesis or upper motor neuron signs, which is one of the major neurological deficits in APBD patients.

[0141] The compound GHF-201 was administered to APBD model mice as a 5% DMSO solution by IV injection at a dose of 250 mg / kg twice weekly (equivalent to a daily dose of 70 mg / kg). Daily injections were avoided due to the relatively long treatment period (6 months), which could potentially lead to excessive scarring. Following positive results in the mouse model, the compound GHF-201 was administered to two APBD patients as part of a 3-day oral dose-escalation study, during which clinical safety and pharmacokinetic profiles were determined.

[0142] For three consecutive days, the following regime was followed: before administration and 0.5, 1, 1.5, 2, and 4 hours after administration. Compound GHF-201 was administered in the formulation LDS-C, 7.15% compound GHF-201. The dose of the active ingredient was 170 mg on the first day, 255 mg on the second day, and 340 mg on the third day.

[0143] After blood collection, plasma was separated by centrifugation (up to 1 hour after collection) at 2000 g for 10 minutes at 4°C. Compound GHF-201 levels in plasma (supernatant) samples were determined by LC-mass spectrometry (LC-MS / MS) using a Sciex Triple Quad™ 5500 mass spectrometer. 100 μl of plasma was collected and extracted in 500 μl of acetonitrile solution (acetonitrile / water 1:1 v / v). A calibration curve was generated using 0.1, 10, 100, and 1000 ng / ml of compound GHF-201 in a 1 mg / ml solution of 4-tert-butyl-2-(4H-1,2,4-triazol-4-yl)phenol (ChemBridge) as the internal standard (IS). The samples were dissolved in 1 mg / ml IS solution, and 0-1000 ng / ml of compound GHF-201 was added to prepare a calibration curve from which the plasma levels of compound GHF-201 were determined.

[0144] The pharmacokinetic profile of patient 1 is shown in comparison with the mouse model profile in Figures 9A-9C, and the comparative pharmacokinetic profile of patient 2 is shown in Figures 10A-10C.

[0145] According to the FDA human equivalent dose calculation, the mouse dose can be converted to a human dose by dividing by 12. The Cmax in mice was 4,700 ng / mL (Figures 9B and 10B). Dividing by 12, 4,700 ng / mL becomes 392 ng / mL. The Cmax in patient 1 was 894 ng / mL (Figure 9A) and 883 ng / mL in patient 2 (Figure 10A). In mice, the administered dose was found to produce a significant therapeutic effect. Therefore, we hypothesized that a human-equivalent, or higher, dose in patients would be at least as effective as the equivalent dose in mice. Another assumption was that the Cmax observed after administration of a therapeutic dose would correlate with biological activity, similar to the administered dose, and that these results could predict therapeutic efficacy in APBD patients.

[0146] Additionally, administration of the LDS-C formulated compound to patients was observed to delay the time to Cmax, suggesting a "sustained-release" mode of delivery. The time to Cmax increased with increasing administered dose. Another interesting observation was that pre-dose levels of compound GHF-201 in plasma increased with each successive day (Figures 9C, 10C), suggesting that residual doses remained in the patient's plasma before the administration of the next dose the following day, and that these doses accumulated.

[0147] Importantly, the area under the curve (AUC) correlated with the level of administered compound GHF-201 (Figures 11A-11B). This observation indicates that the body's exposure to the drug was positively correlated with the level of administered drug. AUC was calculated as a trapezoidal sum rather than using integration and extrapolation to infinite time.

[0148] Figure 12A shows muscle strength grades (before treatment and after 10 months of treatment) for patient 1 (Figure 12A), performed according to the method described in Kleyweg et al. 1991. As can be seen, treatment resulted in a significant increase in muscle strength in all muscles measured.

[0149] Figures 12B-12C show plasma neurofilament light chains for Patient 1 (Figure 12B) and Patient 2 (Figure 12C), who were treated with a formulation of the compound GHF-201. NFL levels were obtained by analyzing patient plasma samples using a Simoa instrument and the NF-Light v2 Advantage HD-X kit for determination of human light chain neurofilament protein.

[0150] The reduction of neurofilament light chain level indicates the amelioration of neurodegeneration.Neurofilament light chain is considered to be a systemic biomarker for the degree of neurodegeneration in some neurodegenerative disorders, such as ALS and AD.Administering GHF-201 formulated with the formulation of this disclosure shows a significant reduction of neurofilament light chain, suggesting the amelioration of the degree of neurodegeneration.

[0151] Further Exemplary Formulations A blank formulation was prepared by weighing all the ingredients according to Table 7 and mixing them at 40-60°C.

[0152] The compound of formula (II), herein designated as compound GHF-205, was used for the additional formulation: [ka] [Table 11]

[0153] The compound GHF-205 was solubilized in the formulations at various concentrations (2.5 and 5.0 wt%). The formulations were tested for appearance, refractive index, and droplet size as detailed in Table 8. [Table 12]

[0154] SD-NMR characterization was also performed on LDS-C(205) loaded with 2.5 and 5 wt % GHF-205 in concentrated form, as detailed in Table 9. [Table 13]

[0155] In general, it can be seen that cosurfactants in concentrated systems diffuse faster (have higher mobility) than surfactants. This suggests that the hydrophilic surfactant and lipophilic components form the main building blocks of the structure's interface in the concentrated form; once diluted, the lipophilic components are no longer needed to stabilize the interface (because their mobility is higher in the diluted form compared to the concentrated form). Thus, in the concentrated form, the lipophilic components are located at the interface and are essential for building the droplet structure; upon dilution, the lipophilic components move away from the interface and are closer to the external phase. This indicates that upon oral ingestion, GHF-205 can migrate from the formulation due to a conformational change.

Claims

1. 1. A pharmaceutical formulation for oral delivery of a compound of formula (I) or a pharmaceutically acceptable salt, isomer, or tautomer thereof, comprising: 【Chemical 1】 【Chemistry 2】 represents a single or double bond, n and m are integers, each independently 1, 2, or 3; R and R １ are each independently hydrogen or absent; R ２ , R ３ , R ４ , R ５ , R ６ , R ７ , R ８ and R ９ are each hydrogen or each independently selected from alkyl, cycloalkyl, alkoxy, hydroxy, thiohydroxy, thioalkoxy, aryloxy, thioaryloxy, amino, nitro, halo, trihalomethyl, cyano, amido, carboxy, sulfonyl, sulfoxy, sulfinyl, and sulfonamido, each further substituted or unsubstituted; and One of X and Y is S and the other of X and Y is C; provided that when X is S, then R ９ is absent and when Y is S, R ５ is not present; The formulation comprises: a) the compound of formula (I) or a pharmaceutically acceptable salt, isomer or tautomer thereof; b) at least one hydrophilic surfactant in a total amount ranging from about 10 wt % to 50 wt %; c) at least one solvent in a total amount of at least about 20 wt %; d) at least one co-surfactant; and e) at least one oil in an amount of 0% to about 5% by weight of the formulation; Including, The pharmaceutical formulation, wherein the weight ratio of said at least one solvent to said at least one hydrophilic surfactant ranges from about 1.25:1 to about 1:

3.

2. The compound of formula (I) is a compound of formula (I') or a pharmaceutically acceptable salt, isomer or tautomer thereof, 【Chemistry 3】 【Chemistry 4】 represents a single or double bond, and n, m, R, and R １ , R ２ , R ３ , R ４ , R ５ , R ６ , R ７ and R ８ 2. The formulation of claim 1, wherein each of is as defined in claim 1.

3. 3. The formulation of claim 1, wherein the total amount of the at least one solvent and the at least one co-surfactant in the formulation is at least 45 wt%.

4. The formulation according to any one of claims 1 to 3, wherein the total amount of the at least one solvent and the at least one co-surfactant in the formulation is at least 50 wt%.

5. A formulation according to any one of claims 1 to 4 which is devoid of water.

6. 6. The formulation of any one of claims 1 to 5, wherein the at least one hydrophilic surfactant is selected from ethoxylated fatty acids, ethoxylated castor oil and its hydrogenated derivatives, polysorbates, ethoxylated alkyl ethers, ethoxylated monoglycerides, polyglycerol esters and sucrose esters, and combinations thereof.

7. 7. The formulation of claim 6, wherein the formulation comprises at least one first hydrophilic surfactant selected from ethoxylated castor oil and its hydrogenated derivatives, and at least one second hydrophilic surfactant selected from polysorbates and ethoxylated monoglycerides.

8. 8. A formulation according to any one of claims 1 to 7, wherein the weight ratio between the total hydrophilic surfactants and the total co-surfactants ranges from about 3:1 to 1:

3.

9. 9. The formulation of any one of claims 1 to 8, wherein the at least one co-surfactant is present in the formulation in a total amount ranging from about 10 wt% to about 45 wt%.

10. 10. The formulation of any one of claims 1 to 9, wherein the at least one co-surfactant is selected from polyethylene glycol 200, polyethylene glycol 400, polyethylene glycol 600, propylene glycol, phosphatidylcholine, diethylene glycol monoethyl ether, and combinations thereof.

11. 11. The formulation of any one of claims 1 to 10, wherein the weight ratio between the total solvent and the total co-surfactant ranges from about 3:1 to 1:

2.

12. The formulation of any one of claims 1 to 11, wherein the at least one oil is selected from ethanol, methanol, n-propanol, benzyl alcohol, and combinations thereof.

13. 13. A formulation according to any one of claims 1 to 12, wherein the at least one oil is present in the formulation at a concentration of up to 4 wt%.

14. 14. The formulation of any one of claims 1 to 13, comprising up to 10 wt% of the compound of formula (I) or a pharmaceutically acceptable salt, isomer or tautomer thereof.

15. The formulation of any one of claims 2 to 14, wherein n and m are 1.

16. R ２ , R ７ and R ８ The formulation of any one of claims 2 to 15, wherein each is methyl.

17. The formulation according to any one of claims 1 to 16, wherein the compound is at least one compound of formula (IA) or (IB): 【Chemistry 5】

18. 1. A pharmaceutical formulation for oral delivery of a compound of formula (II) or a pharmaceutically acceptable salt, isomer, or tautomer thereof, comprising: 【Chemistry 6】 The formulation comprises: a) the compound of formula (II) or a pharmaceutically acceptable salt, isomer or tautomer thereof; b) at least one hydrophilic surfactant in a total amount ranging from about 10 wt % to 70 wt %; c) at least one solvent in a total amount of at least about 15 wt %; d) at least one co-surfactant, and e) at least one oil in an amount of 0% to about 5% by weight of the formulation; Including, A pharmaceutical formulation wherein the weight ratio of said at least one solvent to said at least one hydrophilic surfactant ranges from about 1:1 to about 1:

7.

19. 19. The pharmaceutical formulation of claim 18, comprising a total amount of the at least one hydrophilic surfactant in the range of about 20 wt% to 50 wt%.

20. 20. The pharmaceutical formulation of claim 18 or 19, comprising at least about 20 wt% total amount of said at least one solvent.

21. 21. The pharmaceutical formulation of any one of claims 18 to 20, wherein the weight ratio of the at least one solvent to the at least one hydrophilic surfactant ranges from about 1.25:1 to about 1:

3.

22. 22. A formulation for oral delivery of a compound of formula (I) or a pharmaceutically acceptable salt, isomer or tautomer thereof, comprising nanodroplets of the formulation of any one of claims 1 to 21 dispersed in a continuous phase comprising at least one aqueous diluent.

23. 23. The preparation of claim 22, wherein the nanodroplets have an average droplet size in the range of about 5 nm to 50 nm.

24. 24. The formulation of claim 22 or 23, wherein the at least one aqueous diluent is selected from water, water for injection, saline, dextrose solution, and a buffer.

25. 22. A formulation for oral delivery of a compound of formula (II) or a pharmaceutically acceptable salt, isomer or tautomer thereof, comprising nanodroplets of the formulation of any one of claims 1 to 21 dispersed in a continuous phase comprising at least one aqueous diluent.

26. 26. The preparation of claim 25, wherein the nanodroplets have an average droplet size in the range of about 5 nm to 50 nm.

27. 27. The formulation of claim 25 or 26, wherein the at least one aqueous diluent is selected from water, water for injection, saline, dextrose solution, and a buffer.

28. A formulation according to any one of claims 1 to 21 or a preparation according to any one of claims 22 to 27 for use in treating glycogen storage disease (GSD).

29. 29. The formulation or preparation for use of claim 28, wherein the GSD is GSD Type 0, GSD Type I, GSD Type II, GSD Type III, GSD Type IV, GSD Type V, GSD Type VI, GSD Type VII, GSD Type VII I, GSD Type IX, GSD Type X, GSD Type XI, GSD Type XII, GSD Type XIII, GSD Type XIV, or GSD Type XV.

30. 29. The formulation or preparation for use according to claim 28, wherein the GSD is adult polyglucosan body disorder (APBD), Andersen's disease, Forbes' disease, or Danon's disease.

31. A formulation according to any one of claims 1 to 21 or a preparation according to any one of claims 22 to 27 for use in treating a disease or condition associated with lysosomal storage.

32. 32. The formulation or preparation for use of claim 31 , wherein the disease or condition is selected from Gaucher disease, Fabry disease, Tay-Sachs disease, mucopolysaccharidosis (MPS) disorders, aspartylglucosaminuria, GMI-gangliosidosis, Krabbe (globoid cell leukodystrophy or galactosylceramide lipidosis), metachromatic leukodystrophy, Sandhoff disease, mucolipidosis type II (I-cell disease), mucolipidosis type IIIA (pseudo-Hurler polydystrophy), Niemann-Pick disease types C2 and C1, Danon disease, free sialic acid storage disorders, mucolipidosis type IV, multiple sulfatase deficiency (MSD), metabolic disorders, obesity, type II diabetes, and insulin resistance.

33. A formulation according to any one of claims 1 to 21 or a preparation according to any one of claims 22 to 27 for use in treating a disease or condition associated with autophagy-misregulation.

34. 34. The formulation or preparation for use according to claim 33, wherein the disease or condition is selected from Alzheimer's disease and cancers associated with reduced autophagy activity.

35. 28. A method of treating glycogen storage disease (GSD) comprising administering to a patient in need thereof an effective amount of the formulation of any one of claims 1 to 21 or the preparation of any one of claims 22 to 27.

36. A method of treating a disease or condition associated with lysosomal storage, comprising administering to a patient in need thereof an effective amount of a formulation according to any one of claims 1 to 21 or a preparation according to any one of claims 22 to 27.

37. 28. A method for treating a disease or condition associated with autophagy-misregulation, comprising administering to a patient in need thereof an effective amount of the formulation of any one of claims 1 to 21 or the preparation of any one of claims 22 to 27.

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