Oral delivery of active compounds for treatment of glycogen storage diseases

Abstract

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

Description

Technical Field

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

[0002] Background of the Invention

[0003] Listed below are references believed to be relevant to the background of the presently disclosed subject matter:

[0004] -PCT Patent Application Publication No. WO2018 / 154578

[0005] -PCT patent application serial number PCT / IL2022 / 050187

[0006] -Kapetanovic et al., "Effects of oral dosing paradigms (gavage versus diet) on pharmacokinetics and pharmacodynamics", 2006, Chem Biol Interact, 164, 68-75

[0007] -Kleyweg et al., "Interobserver Agreement In The Assessment Of MuscleStrength And Functional Abilities In Guillain-Barre Syndrome", 1991, Muscle&Nerve, 14, 1103-1109

[0008] Acknowledgment herein of the above references should not be inferred as meaning that these references are in any way relevant to the patentability of the presently disclosed subject matter. Background Art

[0009] Glycogen is a branched polysaccharide composed of glucose units linked by α1-4 glycosidic bonds to form linear chains, which are further linked by α1-6 glycosidic bonds to form branches.

[0010] Glycogen is primarily stored in the liver and muscles, but can also be found at lower levels in the kidneys, heart, and brain. In the liver, it provides a source of energy during fasting, while in the muscles, glycogen is an immediate reserve source of available glucose.

[0011] Glycogen synthesis begins with the autoglycosylation of the glycogenoprotein oligosaccharide primer. Glycogen elongation involves glycogen synthase, which catalyzes the formation of α1-4 glycosidic bonds, and glycogen branching enzyme (GBE), which catalyzes the formation of α1-6 glycosidic bonds and glycogen. The degradation of glycogen occurs through two different pathways, one in the cytosol (called "glycogenolysis"), and the other in the lysosome, called "glycogen autophagy" or "glycoautophagy". The breakdown of glycogen via glycogenolysis involves glycogen phosphorylase (GP) and glycogen debranching enzyme (GDE), while the glycoautophagy mechanism is mediated by acid alpha glycosidase (GAA).

[0012] The importance of glycogen metabolism is not only emphasized in terms of energy maintenance, but it is also associated with several congenital disorders caused by abnormalities in the function of enzymes that control glycogen synthesis, degradation and regulation. These disorders are inherited in an autosomal recessive manner in most cases and are collectively referred to as glycogen storage diseases (GSD). Today, there are 16 subtypes of GSD known to the medical community, including Lafora disease and Danon disease, all of which present with a variety of clinical manifestations.

[0013] Recently, several compounds have been designed for the treatment of GSD and described in PCT patent publication WO2018 / 154578. These compounds have shown good activity in the treatment of GSD. Summary of the invention

[0014] The present disclosure provides formulations for oral delivery of active compounds for treating GSD and treating neurodegenerative disorders or conditions associated with lysosomal storage or autophagy disorders. The formulations of the present disclosure are designed to enable increased loading of active compounds while maintaining high tolerability and improved bioavailability.

[0015] The formulations of the present disclosure are formulated to stabilize the active compound in an anhydrous nanostructure formulation while allowing complete dilution in an aqueous liquid so that once orally administered, the formulation is uniformly dispersed in an aqueous phase (e.g., gastric fluid) to form dispersed nanostructures in which the active compound is captured and stabilized. This capture allows the active compound in the formulation to be stable, and once administered, the active compound is released from the nanostructures to ensure that an efficient dose of the active compound is delivered over time. The inventors have unexpectedly discovered that by utilizing a combination of one or more solvents, one or more co-surfactants, and one or more hydrophilic surfactants that form a substantially hydrophilic delivery system, it is possible to form nanostructures in an anhydrous formulation, as well as to form nanostructures in situ after administration while stabilizing high loads of lipophilic active compounds disclosed herein.

[0016] By one aspect thereof, 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,

[0017]

[0018] represents a single bond or a double bond,

[0019] n and m are integers, each independently 1, 2 or 3;

[0020] R and R 1 each independently is hydrogen or absent;

[0021] R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 and R 9 are each hydrogen or are each independently selected from alkyl, cycloalkyl, alkoxy, hydroxy, thiohydroxy, thioalkoxy, aryloxy, thioaryloxy, amino, nitro, halogen, trihalomethyl, cyano, amide, carboxyl, sulfonyl, sulfonyloxy, sulfinyl and sulfonamide, each of which is further substituted or unsubstituted; and

[0022] 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 does not exist, and when Y is S, then R 5 does not exist;

[0023] Wherein, the preparation comprises:

[0024] a) the compound of formula (I) or a pharmaceutically acceptable salt, isomer or tautomer thereof;

[0025] b) at least one hydrophilic surfactant in a total amount ranging from about 10 wt % to 50 wt %;

[0026] c) at least one solvent in a total amount of at least about 20 wt %;

[0027] d) at least one co-surfactant; and

[0028] e) at least one oil in an amount between 0 wt % and about 5 wt % of the formulation;

[0029] 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.

[0030] In some embodiments, the compound of formula (I) is a compound of formula (I') or a pharmaceutically acceptable salt, isomer or tautomer thereof,

[0031]

[0032] represents a single bond or a double bond,

[0033] n and m are integers, each independently 1, 2 or 3;

[0034] R and R 1 are each independently hydrogen or absent; and

[0035] R 2 , R 3 , R 4 , R 5 , R 6 , R 7 and R 8 each is hydrogen, or

[0036] R 2 , R 3 , R 4 , R 5 , R 6 , R 7 and R 8 Each is independently selected from alkyl, cycloalkyl, alkoxy, hydroxy, thiohydroxy, thioalkoxy, aryloxy, thioaryloxy, amino, nitro, halogen, trihalomethyl, cyano, amide, carboxyl, sulfonyl, sulfonyloxy, sulfinyl and sulfonamide, each of which is further substituted or unsubstituted.

[0037] The formulations disclosed herein are designed for oral delivery of active compounds, i.e., delivery of active compounds by swallowing, to obtain systemic pharmacological effects. The formulations are typically in liquid form and can be administered as liquids, gels, suspensions, or encapsulated in liquid gel or soft gel capsules.

[0038] The formulations of the present disclosure, due to their unique formulation composition, are capable of stably loading the active compound into the formulation at a concentration of at least 0.5 wt %, such as 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.

[0039] According to some embodiments, the formulation comprises up to about 10 wt % of the compound of formula (I) or a pharmaceutically acceptable salt, isomer or tautomer thereof.

[0040] The inventors have discovered that a combination of one or more hydrophilic surfactants, one or more co-solvents, and one or more solvents can load the active compound into the formulation at high levels and stabilize it for long periods of time, while allowing spontaneous formation of nanostructures in anhydrous formulations and when mixed with aqueous liquids (e.g., gastric fluid after administration). In the formulations of the present disclosure, the balance of ingredients allows for high loading and entrapment of lipophilic active agents in a predominantly hydrophilic formulation for long periods of time, thereby allowing for a longer shelf life while minimizing phase separation and / or sedimentation.

[0041] The formulations of the present disclosure are generally in the form of a concentrate, typically a water-free concentrate, which is stable to dilution with an aqueous medium (i.e., the droplet size or phase separation does not increase significantly upon dilution). The concentrate form is stable for a long time, lacks a life-supporting environment for microorganisms, and can be readily diluted in an aqueous medium, as will be further explained below.

[0042] Thus, the formulations disclosed herein are generally free of water. The formulations are designed to allow spontaneous formation of nanostructures, either in concentrate form (ie, in the absence of water) or when mixed with an aqueous liquid (after or for administration), as will be described in further detail below.

[0043] In the formulations of the present disclosure, the delicate balance between surfactant and co-surfactant / solvent imparts physical stability to the formulation (without water) in the presence of high concentrations of pharmaceutically active compounds, as this balance is found to promote the dissolution of the active compound. At the same time, the ratio between surfactant and co-surfactant / solvent permits the dilution capability of the formulation in aqueous liquids, allowing the formation of extremely small droplets less than about 10 nm.

[0044] The term one or more hydrophilic surfactants refers to surfactants having a hydrophilic head group and a lipophilic tail that are capable of arranging into nanostructures in aqueous media. The inventors have discovered that the combination of a hydrophilic surfactant with a co-surfactant and a solvent in a specific range of ratios and total concentrations can spontaneously form stable nanostructures that stabilize the active compound in the formulation in an anhydrous concentrate and dissolve the active compound in the nanostructures when mixed with an aqueous liquid.

[0045] According to some embodiments, the at least one hydrophilic surfactant is selected from the group consisting of ethoxylated fatty acids, ethoxylated castor oil and its hydrogenated derivatives, polysorbates, ethoxylated alkyl ethers, ethoxylated monoglycerides, polyglycerol esters, sucrose esters, and combinations thereof.

[0046] According to some embodiments, the formulation includes at least one first hydrophilic surfactant selected from ethoxylated castor oil and hydrogenated derivatives thereof (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 polyoxyethylene-8 glyceride, lauryl polyoxyethylene 32 glyceride, stearyl polyoxyethylene 32 glyceride, etc.).

[0047] As described, at least one hydrophilic surfactant is present in the formulation in an amount between about 10 wt% and about 50 wt%. According to some embodiments, the total amount of at least one hydrophilic surfactant is between about 20 wt% and 50 wt%. According to other embodiments, the total amount of at least one hydrophilic surfactant is between about 25 wt% and 50 wt%.

[0048] The formulation includes at least one co-surfactant. It is understood that a co-surfactant includes any hydrophilic, lipophilic or amphiphilic agent, different from one or more of the hydrophilic surfactants, which helps (together with the surfactant) reduce the interfacial tension between the oil phase and the aqueous phase to almost zero (or zero), allowing the formation of thermodynamically stable nanostructures. Thus, the combination of surfactant and co-surfactant allows the formulation to be kinetically and thermodynamically stable.

[0049] 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 (such as phosphatidylcholine), diethylene glycol monoethyl ether, and combinations thereof.

[0050] According to some embodiments, 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, 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%.

[0051] According to some embodiments, the weight ratio between the total hydrophilic surfactant and the total co-surfactant in the formulation ranges from about 3:1 to about 1:3. According to some embodiments, the weight ratio between the total hydrophilic surfactant and the total co-surfactant in the formulation is 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:, 2.0:1, 1.9:1, 1.8:1, 1.7:1, 1.6:1, 1.5:1, 1.4:1, 1 .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 about 1:3.

[0052] According to some embodiments, the weight ratio of the total hydrophilic surfactant to the total co-surfactant in the formulation ranges from about 2.5:1 to about 1:1.5. According to yet other embodiments, the weight ratio of the total hydrophilic surfactant to the total co-surfactant in the formulation ranges from about 2:1 to about 1:1.

[0053] The formulation also includes a relatively large amount (usually at least 20 wt%) of at least one solvent. The solvent is an organic solvent, typically polar, which is at least partially miscible with water and is suitable for facilitating dissolution of the active compound in the formulation and in the nanostructures.

[0054] 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 35 wt%.

[0055] According to some embodiments, the at least one solvent is selected from ethanol, methanol, n-propanol, benzyl alcohol, and combinations thereof.

[0056] 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%.

[0057] 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 between the total amount of solvent and the total amount of co-surfactant ranges from about 2: 1 to about 1: 1.5.

[0058] 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.

[0059] As described, 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.

[0060] 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.

[0061] The formulation comprises between 0 wt% and about 5 wt% oil. According to some embodiments, at least one oil is present in the formulation at a concentration of no more than 4 wt%. According to some embodiments, the formulation contains no oil.

[0062] The term oil refers to a lipophilic agent that is immiscible in water and is capable of forming distinct domains when introduced into an aqueous liquid. In some embodiments, the at least one oil is selected from short chain triglycerides and medium chain triglycerides.

[0063] In some embodiments, the formulation may further include various additives approved for pharmaceutical use, such as pH adjusters and buffers, neutralizers, softeners, humectants, preservatives, antioxidants, taste masking agents, flavoring agents, sweeteners, flavoring additives, and any other suitable inactive pharmaceutical additives.

[0064] The formulations disclosed herein are designed as pharmaceutical formulations for oral delivery of a compound of formula (I) or a pharmaceutically acceptable salt, isomer or tautomer thereof.

[0065] According to some embodiments, n and m in formula (I) are 1.

[0066] According to some embodiments, R in formula (I) 2 , R 7 and R 8 Each is a methyl group.

[0067] According to some embodiments, the compound of formula (I) is at least one compound of formula (IA) or (IB):

[0068]

[0069] According to other embodiments, the formulation comprises two or more compounds of formula (I).

[0070] As used herein, alkyl, alkenyl, alkynyl carbon chain (if not otherwise specified) contains 1 to 20 carbons, and may be straight or branched. In certain embodiments, the alkenyl carbon chain of 2 to 20 carbons contains 1 to 8 double bonds, and in certain embodiments, the alkenyl carbon chain of 2 to 16 carbons contains 1 to 5 double bonds. In certain embodiments, the alkynyl carbon chain of 2 to 20 carbons contains 1 to 8 triple bonds, and in certain embodiments, the alkynyl carbon chain of 2 to 16 carbons contains 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).

[0071] C 1-6 Alkyl is understood to include any straight or branched chain alkyl moiety having 1, 2, 3, 4, 5 or 6 carbon atoms.

[0072] Cycloalkyl refers to a saturated monocyclic or polycyclic ring system having 3 to 10 carbon atoms in certain embodiments and 3 to 6 carbon atoms in other embodiments; cycloalkenyl and cycloalkynyl refer to monocyclic or polycyclic ring systems comprising at least one double bond and at least one triple bond, respectively. In certain embodiments, cycloalkenyl and cycloalkynyl groups may contain 3 to 10 carbon atoms, in further embodiments, cycloalkenyl groups contain 4 to 7 carbon atoms, and in further embodiments, cycloalkynyl groups contain 8 to 10 carbon atoms. The ring systems of cycloalkyl, cycloalkenyl, and cycloalkynyl groups may consist of one ring or two or more rings, which may be connected together in a fused, bridged, or spirally connected manner.

[0073] Heterocyclyl refers to a monocyclic or polycyclic non-aromatic ring system, in one embodiment having from 3 to 10 members, wherein one or more atoms in the ring system is a heteroatom, ie, an element other than carbon, including but not limited to nitrogen, oxygen or sulfur.

[0074] Alkoxy refers to -O-alkyl or -O-cycloalkyl as defined herein; thioalkoxy refers to -S-alkyl or -S-cycloalkyl as defined herein.

[0075] Aryl refers to an aromatic monocyclic or polycyclic carbon group containing 5 to 19 carbon atoms, ie having a conjugated π electron system.

[0076] Heteroaryl refers to a monocyclic or polycyclic aromatic ring system, which in certain embodiments has from about 5 to about 19 members, wherein one or more atoms in the ring system is a heteroatom, i.e., an element other than carbon, including but not limited to nitrogen, oxygen or sulfur.

[0077] Aryloxy refers to -O-aryl or -O-heteroaryl as defined herein; thioaryloxy refers to -S-aryl or -S-heteroaryl as defined herein.

[0078] Hydroxyl refers to the -OH group.

[0079] Thiol (or thiol) refers to a -SH group.

[0080] Amino refers to a primary, secondary or tertiary amine (-NR'R", R' and R" are independently hydrogen, alkyl, cycloalkyl, alkenyl, aryl, heteroaryl or heterocyclyl, as defined herein), wherein the point of attachment is through the nitrogen atom substituted with a C1-C6 straight or branched chain alkyl. For tertiary amines, the substituents may be the same or different.

[0081] Nitro refers to the -NO2 group.

[0082] Halogen or (or halogen or halide) refers to F, Cl, Br or I. Haloalkyl refers to an alkyl group in which one or more hydrogen atoms are replaced by halogen. Such groups include, but are not limited to, trihalomethyl.

[0083] Cyano refers to a -C≡N group.

[0084] Amide refers to the divalent group -C(O)NH2.

[0085] Carboxyl refers to a -C(O)-OR' group, wherein R' is hydrogen, alkyl, cycloalkyl, alkenyl, aryl, heteroaryl or heterocyclyl, as defined herein.

[0086] Sulfonyl refers to a -S(O)2-R' group, wherein R' is hydrogen, alkyl, cycloalkyl, alkenyl, aryl, heteroaryl or heterocyclyl, as defined herein.

[0087] Sulfinyl refers to a -S(O)-R' group, wherein R' is hydrogen, alkyl, cycloalkyl, alkenyl, aryl, heteroaryl or heterocyclyl, as defined herein.

[0088] Sulfonamide refers to a -S(O)2-NR'R" group, R' and R" are independently hydrogen, alkyl, cycloalkyl, alkenyl, aryl, heteroaryl or heterocyclyl, as defined herein.

[0089] As used herein, the term one or more pharmaceutically acceptable salts means those salts of the disclosed compounds, which are safe and effective for mammalian pharmaceutical use, and have the desired biological activity. Pharmaceutically acceptable salts include salts of acidic or basic groups present in the disclosed compounds. Pharmaceutically acceptable acid addition salts include but are not limited to hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate, lactate, salicylate, citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate, fumarate, gluconate, glucuronate, sucrose, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate (i.e., 1,1'-methylene-bis-(2-hydroxy-3-naphthoate)) salts. Certain compounds of the present 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.

[0090] The compounds described herein include one or more chiral atoms, or are otherwise capable of existing as isomers, such as two enantiomers or as two or more diastereomers. Thus, the compounds may include isomeric mixtures as well as purified isomers or enantiomerically enriched mixtures. In addition, the compounds may include diastereomeric mixtures as well as purified stereoisomers or diastereoisomerically enriched mixtures. It is also noted that the compounds may form tautomers, isolated or any mixture thereof.

[0091] According to another aspect of the present disclosure, a pharmaceutical formulation for oral delivery of a compound of formula (II) or a pharmaceutically acceptable salt, isomer or tautomer thereof is provided.

[0092]

[0093] Wherein, the preparation comprises:

[0094] a) the compound of formula (II) or a pharmaceutically acceptable salt, isomer or tautomer thereof,

[0095] b) at least one hydrophilic surfactant in a total amount ranging from about 10 wt % to 70 wt %,

[0096] c) at least one solvent in a total amount of at least about 15 wt %;

[0097] d) at least one co-surfactant, and

[0098] e) at least one oil in an amount between 0 wt % and about 5 wt % of the formulation,

[0099] 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.

[0100] According to some embodiments, the formulation may include a mixture of at least one compound of formula (I) and a compound of formula (II).

[0101] According to some embodiments, the formulation comprises up to about 10 wt % of the compound of formula (II) or a pharmaceutically acceptable salt, isomer or tautomer thereof.

[0102] According to some embodiments, the at least one hydrophilic surfactant is selected from: ethoxylated fatty acids, ethoxylated castor oil and hydrogenated derivatives thereof, polysorbates, ethoxylated alkyl ethers, ethoxylated glycerol monoesters, 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 hydrogenated derivatives thereof (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 glycerol monoesters (caprylocaproyl polyoxyethylene-8 glyceride, lauryl polyoxyethylene 32 glyceride, stearyl polyoxyethylene 32 glyceride, etc.).

[0103] According to some embodiments, the total amount of the at least one hydrophilic surfactant is in the range of about 20wt% to 70wt% of the compound (II) formulation. According to other embodiments, the total amount of the at least one hydrophilic surfactant is in the range of about 30wt% to 70wt%. According to some embodiments, the formulation of the compound of formula (II) includes the at least one hydrophilic surfactant in a total amount in the range of about 20wt% to 50wt%.

[0104] 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.

[0105] According to some embodiments, at least one co-surfactant is present in a total amount ranging from about 8 wt% to about 45 wt% in the formulation of the compound of formula (II). According to other embodiments, at least one co-surfactant is present in a total amount ranging from about 8 wt% to about 35 wt% in the formulation of the compound of formula (II).

[0106] According to some embodiments, the weight ratio between the total hydrophilic surfactant and the total co-surfactant in the formulation of the compound of formula (II) ranges from about 7: 1 to about 1: 3. According to some embodiments, the weight ratio between the total hydrophilic surfactant and the total co-surfactant in the formulation ranges from about 7: 1 to about 1: 1, for example, 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.

[0107] The formulation of the compound of formula (II) also includes a relatively large amount (usually at least 15wt%) of at least one solvent. According to some embodiments, the formulation of the compound of formula (II) includes the at least one solvent in a concentration range of about 15wt% to about 45wt%. According to some embodiments, the formulation of the compound of formula (II) includes the at least one solvent in a concentration range of about 15wt% to about 35wt%.

[0108] According to some embodiments, the at least one solvent is selected from ethanol, methanol, n-propanol, benzyl alcohol, and combinations thereof.

[0109] 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.

[0110] 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%.

[0111] Formulations of compounds of formula (II) include between 0 wt% and about 5 wt% oil, preferably up to 2 wt% oil, and even more preferably contain no oil.

[0112] In some embodiments, the 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, neutralizers, softeners, humectants, preservatives, antioxidants, taste masking agents, flavoring agents, sweeteners, flavoring additives, and any other suitable inactive pharmaceutical additives.

[0113] The formulations of the compounds of formula (I), (I') and / or (II) disclosed herein are designed as pharmaceutical formulations for oral delivery of the compounds of formula (I), (I') and / or (II) or pharmaceutically acceptable salts, isomers or tautomers thereof.

[0114] As described, the formulations of the compounds of formula (I), (I') and / or (II) of the present disclosure are designed to be stable (thermodynamically and kinetically) over long periods of time. The formulations of the present disclosure form nanostructures both in concentrate form (i.e., without water) and once mixed with an aqueous liquid. The nanostructures stabilize and capture the active compound to allow it to be contained in the formulation prior to dilution with an aqueous liquid and to be released from the nanostructures after administration (i.e., after dilution).

[0115] The fine balance between one or more hydrophilic surfactants, one or more co-surfactants and one or more solvents allows the spontaneous formation of nanostructures in which the active compound is dissolved and stable. The combination of one or more hydrophilic surfactants, one or more co-surfactants and one or more solvents facilitates complete coverage of the interface between the nanostructure and the aqueous diluent at high water dilution of the formulation. In other words, the combination of hydrophilic surfactants, co-surfactants and solvents changes the effective critical packing parameter (ECPP) of the interface, facilitating control of the hydrophilicity / hydrophobicity of the surfactant depending on the amount of water, thereby improving the stability of the nanostructure.

[0116] The formulations of the present disclosure can be administered as is, i.e., in the form of a concentrate that can be easily diluted in situ by gastric fluid after administration. Alternatively, the formulations can be administered in a diluted form by diluting the formulation with one or more aqueous diluents prior to administration.

[0117] Therefore, according to another aspect thereof, 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 the preparation, dispersed in a continuous phase comprising at least one aqueous diluent, the preparation comprising the compound of formula (I) disclosed herein or a pharmaceutically acceptable salt, isomer or tautomer thereof.

[0118] On the other hand, a preparation for oral delivery of a compound of formula (I') or a pharmaceutically acceptable salt, isomer or tautomer thereof is provided, the preparation comprising nanodroplets of the preparation dispersed in a continuous phase comprising at least one aqueous diluent, the preparation comprising the compound of formula (I') or a pharmaceutically acceptable salt, isomer or tautomer thereof.

[0119] On the other hand, a preparation for oral delivery of a compound of formula (II) or a pharmaceutically acceptable salt, isomer or tautomer thereof is provided, the preparation comprising nanodroplets of the preparation dispersed in a continuous phase comprising at least one aqueous diluent, the preparation comprising the compound of formula (II) or a pharmaceutically acceptable salt, isomer or tautomer thereof.

[0120] It is generally noted that, unless specifically stated otherwise, the term formulation is used to denote an anhydrous composition (ie, a concentrate form), whereas the term preparation is intended to refer to a diluted form of a formulation.

[0121] Nanodroplets (or nanostructures) are droplets composed of a formulation that captures and stabilizes a pharmaceutically active compound. The nanostructures are typically in the form of liquid droplets with an average diameter of up to 50 nm (nanometers) in which a hydrophilic surfactant and a co-surfactant form an interface between a continuous phase of solvent (when in the form of an anhydrous concentrate) or a continuous aqueous phase and an oil core. Without being bound by theory, the active compound is located at the interface such that at least some of the active compound is physically captured between the heads of the co-surfactant to stabilize it within the nanostructure.

[0122] According to some embodiments, the nanodroplets have an average droplet size ranging between about 5 nm and 50 nm.

[0123] The term mean size refers to the arithmetic mean of the measured droplet diameters. When the droplets are not spherical, the calculation of the mean size is based on an equivalent sphere of the largest dimension of the particle.

[0124] According to some embodiments, the nanodroplets are substantially monodisperse. Due to the size of their monodisperse submicron nanostructures, the formulations and preparations are generally transparent (or substantially transparent), thereby maintaining their transparency for a long time. This allows for easy detection of changes in the stability of the formulation and / or preparation (because phase separation, biologically active precipitation and / or droplet coalescence will result in detectable turbidity).

[0125] According to some embodiments, the at least one aqueous diluent is selected from water, water for injection, saline, dextrose solution and buffered solution.

[0126] According to another aspect, there is provided a formulation or preparation as disclosed herein for use in the treatment of glycogen storage disease (GSD).

[0127] According to another 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 as disclosed herein.

[0128] According to some embodiments, GSD is associated with glycogen branching enzyme deficiency. Glycogen branching enzyme deficiency refers to a disease or condition characterized by the deposition, accumulation or aggregation of polyglucose bodies in the muscles, nerves and / or other tissues of the body.

[0129] 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.

[0130] According to other embodiments, the GSD is adult polyglucose body disorder (APBD), Anderson's disease, Forbes disease, or Danon disease.

[0131] According to another aspect of the present disclosure, there is provided a formulation or preparation as disclosed herein for use in treating a disease or disorder associated with lysosomal storage.

[0132] According to yet another aspect, the present disclosure provides a method of treating a disease or disorder associated with lysosomal storage, comprising administering to a patient in need thereof an effective amount of a formulation or preparation as disclosed herein.

[0133] Lysosomal storage disorders (LSDs) are a group of inherited diseases characterized by lysosomal dysfunction and neurodegeneration. These disorders are usually caused by single gene defects, primarily in specific enzymes required for the normal breakdown of glycosaminoglycans (GAGs). Such defects prevent cells from excreting carbohydrate residues, causing them to accumulate in lysosomes within the cell, leading to disruption of normal cellular function. Exemplary lysosomal storage disorders are sphingolipid storage disorders, ceramidases (e.g., Farber disease, Krabbe disease), galactosialidase storage disorders, gangliosidosis including α-galactosidase (e.g., Fabry disease (α-galactosidase A), Schindler disease (α-galactosidase B)), β-galactosidase (e.g., GM1 gangliosidosis, GM2 gangliosidosis, Sandhoff disease, Tay-Sachs disease), glucocerebrosidosis (e.g., Gaucher disease (type I, II, III), sphingomyelinase (e.g., lysosomal acid lipase deficiency, Niemann-Pick disease), sulfatide storage disorders (e.g., metachromatic leukodystrophy, multiple sulfatase deficiency), mucopolysaccharidosis (e.g., type I (MPS I (Hurler syndrome, Scheher syndrome, Hurler-Schwein syndrome), II (Hunter syndrome), III (Sanfilippo syndrome), IV (Morchio syndrome), VI (Maroto-Rami syndrome), VII (Seeley syndrome), IX (hyaluronidase deficiency), mucolipidosis (e.g., type I (sialidosis), type II (I-cell disease), type III (pseudo-Hurler polydystrophy / phosphotransferase deficiency), type IV (mucolipidase 1 deficiency)), lipidosis (e.g., Niemann-Pick disease), neuronal ceroid lipofuscinosis (e.g., type 1 Santavuori-Halty disease / infantile NCL (CLN1 PPT1)), type 2 juvenile neuronal ceroid lipofuscinosis / late infantile NCL (CLN2 / LINCL) TPP1), Barton-Spielmaier-Voigt disease type 3 / juvenile NCL (CLN3), Kufs disease type 4 / adult NCL (CLN4), Finnish variant type 5 / late infantile (CLN5), late infantile variant type 6 (CLN6), CLN7 type 7, Northern epilepsy type 8 (CLN8), Turkish late infantile type 8 (CLN8), German / Serbian late infantile type 9, congenital cathepsin D deficiency type 10 (CTSD), Wolman disease, oligosaccharidosis (e.g., α-mannosidosis, β-mannosidosis, aspartylglucosaminuria, fucosidosis), lysosomal transport disorders (e.g., cystinosis, pycnodystrophy, Sara disease / sialidosis, infantile free sialic acid storage disease), Pompe disease type II, Danon disease type LIB), cholesterol ester storage disease, etc.

[0134] According to some embodiments, the disease or disorder associated with lysosomal storage is selected from Gaucher disease, Fabry disease, Tay-Sachs disease, mucopolysaccharidosis (MPS) disorders, aspartylglucosaminuria, GMI-gangliosidosis, Krabbe disease (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 type 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.

[0135] According to another aspect, there is provided a formulation or preparation as disclosed herein for use in treating a disease or disorder associated with dysregulation of autophagy.

[0136] According to yet another aspect, there is provided a method for treating a disease or condition associated with dysregulated autophagy, comprising administering to a patient in need thereof an effective amount of a formulation or preparation as disclosed herein.

[0137] Autophagy refers to a catabolic process involving the degradation of the cell's own components, such as long-lived proteins, protein aggregates, organelles, cell membranes, organelle membranes, and other cellular components. The mechanism of autophagy may include: (i) the formation of a membrane around the target area of ​​the cell, separating the contents from the rest of the cytoplasm, (ii) the resulting vesicles fuse with lysosomes, and then the vesicle contents are degraded. Diseases associated with autophagy disorders can be 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, alpha-1 antitrypsin deficiency, dentate nucleus rubrum pallidum Lewy body atrophy, frontotemporal dementia, progressive supranuclear palsy, X-linked spinobulbar muscular atrophy, and neuronal intranuclear hyaline inclusion disease. Also included are diseases or disorders such as cancer, cardiovascular, nervous system degeneration, metabolism, lung, kidney, infectious, musculoskeletal and ocular disorders, wherein the induction of autophagy can contribute to delaying the onset, slowing down, stopping or reversing the progress of one or more symptoms associated with the disease or disorder. Diseases associated with autophagy dysregulation also include cancer, for example, any cancer, wherein 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 further means to include mental illness or disorders, such as any mental illness or disorder, wherein the induction of autophagy will contribute to delaying the onset, slowing down, stopping or reversing the progress of one or more symptoms associated with the mental illness or disorder. In one embodiment, the mental illness or disorder is selected from schizophrenia and bipolar disorder.

[0138] According to some embodiments, the disease or disorder associated with dysregulated autophagy is selected from Alzheimer's disease and a cancer associated with reduced autophagic activity.

[0139] According to some other embodiments, the disease or disorder is a neurodegenerative disease, for example 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.

[0140] As is well known, the effective amount for the purposes herein can be determined by considerations known in the art. The amount must effectively achieve the desired therapeutic effect, which depends in particular on the type and severity of the disease to be treated and the treatment regimen. The effective amount is usually determined in appropriately designed clinical trials (dose range studies), and those skilled in the art will know how to correctly conduct such tests to determine the effective amount. As is well known, the effective amount depends on a variety of factors, including a variety of pharmacological parameters such as half-life in vivo, undesirable side effects (if any), factors such as age and gender.

[0141] 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 a disease, prevent the manifestation of such symptoms before they occur, slow the progression of a disease, slow the worsening of symptoms, enhance the onset of remission, slow the irreversible damage caused by the progressive chronic phase of a disease, delay the onset of the progressive phase, lessen the severity or cure the disease, increase survival or faster recovery, or prevent the occurrence of the disease, or a combination of two or more of the foregoing.

[0142] The term subject means a mammal, human or non-human.

[0143] As used herein, the term about is intended to include ±10% deviation from the specifically mentioned parameter value (such as temperature, concentration, etc.).

[0144] Unless expressly stated otherwise, all concentrations disclosed herein are provided as weight percent (wt %), expressed as a percentage of the weight of the formulation or preparation, as the case may be.

[0145] Whenever a numerical range is indicated herein, it is meant to include any cited numeral (fractional or integer) within the indicated range. The phrases ranging between a first indicated numeral and a second indicated numeral and "ranging" from a first indicated numeral "to" a second indicated numeral are used interchangeably herein and are meant to include the first indicated numeral and the second indicated numeral and all fractions and integers therebetween.

[0146] Unless the context requires otherwise, the word comprise and variations such as "comprises" and "comprising", will be understood to imply the inclusion of stated integers or steps or groups of integers or steps but not the exclusion of any integer or step or group of integers and steps.

[0147] The term ...at least one ... when applied to any component of a formulation is to be understood to include one, two, three, four or even more different occurrences of that component in the formulation or preparation.

[0148] It should be understood that certain features of the invention described in the context of separate embodiments for the sake of clarity may also be provided in combination in a single embodiment. Conversely, various features of the invention described in the context of a single embodiment for the sake of brevity may also be provided separately or in any suitable subcombination or in any other described embodiment of the invention in an appropriate manner. Certain features described in the context of various embodiments should not be considered essential features of those embodiments unless the embodiment cannot function without those elements. BRIEF DESCRIPTION OF THE DRAWINGS

[0149] In order to better understand the subject matter disclosed herein and to illustrate how it can be implemented in practice, embodiments will now be described, by way of non-limiting example only, with reference to the accompanying drawings, in which:

[0150] Figures 1A-1F are pictures of exemplary formulations according to some examples of the present disclosure.

[0151] Figure 2A-2C Droplet size (Z-average) (●) and PDI (■) measurements are shown as a function of compound GHF-201 concentration in the formulation: 8CS-RG ( Figure 2A )、LDS-C( Figure 2B )、LDS-600( Figure 2C ).

[0152] Figures 3A-3C The viscosity changes with the concentration of compound GHF-201 in the formulation: 8CS-RG ( Figure 3A )、LDS-C( Figure 3B )、LDS-600( Figure 3C ).

[0153] Figure 4A-4B The refractive index RI ( Figure 4A ) and normalized RI( Figure 4B ) Viscosity changes with the concentration of compound GHF-201 in the formulation: 8CS-RG (●), LDS-C (■), LDS-600 (▲).

[0154] Figures 5A-5L is the LUMiFuge test result of an exemplary formulation loaded with compound GHF-201: 8CS-RG carrier ( 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 vector( Figure 5E )、LDS-C 2.5% GHF-201( Fig. 5F )、LDS-C 5.0% GHF-201( Figure 5G )、LDS-C 7.5%GHF-201( Figure 5H )、LDS-600 carrier( Fig.5I )、LDS-600 2.5% GHF-201( Figure 5J )、LDS-600 5.0% GHF-201( Figure 5K )、LDS-600 7.5% GHF-201( Figure 5L ).

[0155] Figures 6A-6C This is a photo of the physical stability test of formulation LDS-C, 7.15% compound GHF-201: t = 0 ( Fig. 6A ), 12 months at 25℃ ( Figure 6B ), and after 12 months at 40°C ( Figure 6C ).

[0156] Figures 7A-7C The DLS analysis results of the physical stability test of the formulation LDS-C, 7.15% compound GHF-201 are shown: at t = 0 ( Fig. 7A ), 12 months at 25℃ ( Figure 7B ), and after 12 months at 40°C ( Figure 7C ).

[0157] Figures 8A-8B The results of tissue penetration test of compound GHF-201 in APBD model mice are shown: Unformulated ( Fig. 8A ) and formulated into LDS-C preparation, 7.5wt% ( Figure 8B ).

[0158] Figures 9A-9C is the pharmacokinetic profile of compound GHF-201, which was orally administered to APBD patient 1 ( Fig.9A) and administered intravenously at 250 mg / kg to n=3 APBD model mice ( Fig. 9B ). The doses of active ingredient on days 1, 2, and 3 were 170 mg, 255 mg, and 340 mg, respectively. The circled values ​​in A are Fig. 9C Medium zoom.

[0159] Figures 10A-10C is the pharmacokinetic profile of compound GHF-201, which was orally administered to APBD patients 2 ( Fig. 10A ), and 250 mg / kg was intravenously administered to n=3 APBD model mice ( Fig. 10B ). The doses of active ingredient on days 1, 2 and 3 were 170 mg, 255 mg and 340 mg respectively. Fig. 10C Medium zoom.

[0160] Figures 11A-11B Area under the curve (AUC) for patients 1 and 2 on consecutive days in relation to the dose administered, respectively.

[0161] Figures 12A-12C The muscle strength level of patient 1 (before treatment and after 10 months of treatment) is shown ( Fig. 12A ), and patient 1( Fig. 12B ) and patient 2 ( Fig. 12C ) in plasma neurofilament light chains in patients treated with a formulation of the compound GHF-201. DETAILED DESCRIPTION

[0162] Exemplary Formulations

[0163] Prepare the blank formulation by weighing all components according to Table 1 and mixing them at 40-60°C.

[0164] The compound of formula (IA) is selected as an exemplary compound, which is referred to herein as compound GHF-201 (or alternatively as compound A):

[0165]

[0166] Compound GHF-201 was then dissolved into the formulation at various concentrations (2.5, 5.0, 7.5 wt%, weight percentage based on the weight of the empty formulation).

[0167] Table 1: Exemplary Formulation (without Compound GHF-201)

[0168]

[0169] Table 1 (continued): Exemplary Formulation (without Compound GHF-201)

[0170]

[0171]

[0172] Physical characteristics

[0173] Formulations such as those in Table 1 above do not contain water and are completely dilutable in aqueous liquids. During dilution, the formulations form nanostructures with an average droplet size of approximately 20 nm.

[0174] Selected formulations from Table 1 were analyzed to evaluate their physical characteristics. Figures 1A-1F An exemplary visualization of the appearance of some formulations can be seen in: Undiluted form ( Figures 1A-1C ) and diluted (90% water content) ( Figure 1D-1E ), without compound GHF-201 ("carrier"), and loaded with 2.5wt%, 5wt% and 7.5wt% of compound GHF-201. It can be seen that all formulations, whether in undiluted or diluted form, exhibit high transparency, homogeneity and thermodynamic stability.

[0175] The hydrodynamic radius of the droplets was measured by dynamic light scattering (DLS) using a Nano-ZS Zetasizer (Malvern, UK) at room temperature, with water as the dispersant; the results are shown in Table 2 and Figure 2A-2C shown.

[0176] Table 2: Droplet size and PDI of selected formulations

[0177]

[0178]

[0179] As shown in Table 2 and Figure 2A-2C As can be seen in the , with the increase of the concentration and dissolution of compound GHF-201 in the formulation, the droplets will shrink and the PDI will increase slightly. For the empty formulation (carrier), the average droplet size varies from 13 to 14nm, while for the formulation loaded with compound GHF-201, the droplet sizes of 2.5% and 5-7.5% compound GHF-201 are about 10-11nm and 9-10nm, respectively. The PDI values ​​measured for the carrier are about 0.03-0.06, and for 7.5% compound GHF-201, it is as high as 0.11. Overall, the PDI values ​​of both the carrier and the loaded formulation are very low, indicating a relatively monodisperse system.

[0180] Without being bound by theory, it is hypothesized that the dissolution of compound GHF-201 leads to stronger interactions between the co-surfactant and the solvent at the droplet interface. By increasing the concentration of compound GHF-201, more droplets of smaller size are formed, resulting in a gradual decrease in the average droplet size and an increase in polydispersity.

[0181] To further understand the interaction between compound GHF-201 and the excipients in the formulation, viscosity and NMR measurements were performed.

[0182] Viscosity measurements were performed using an RS6000 rheometer (ThermoScientific) equipped with a C60 / 1 TiL-L12007 cone, operated in rotational mode, with a shear rate range of 0.01000-100.0 1 / s for 6 minutes. All measurements were performed at 25±1°C. Under these conditions, the formulations exhibit Newtonian behavior and, therefore, the formulations can be characterized by their viscosity. The viscosity of the system was calculated based on a linear fit of the rotational shear (τ) versus the shear rate (γ). The slope of the linear fit represents the viscosity. Table 3 and Figures 3A-3C The measured viscosities of preconcentrates for the 8CS-RG, LDS-C and LDS-600 systems are summarized.

[0183] Table 3: Select the viscosity of the formulation

[0184]

[0185]

[0186] like Figures 3A-3C As shown, an increase in the concentration of compound GHF-201 leads to an increase in the viscosity of the formulation. Without being bound by theory, the presence of compound GHF-201 in all tested formulations leads to a stronger interaction between the API and the excipients (most importantly the co-surfactant), which helps the formulation to have greater resistance to applied shear. This suggests that the introduction of compound GHF-201 into the formulation can improve its stability and its resistance to dynamic changes.

[0187] The pH and refractive index of three exemplary formulations were measured. The results are summarized in Table 4. To measure pH, the samples were diluted 10 times with purified water; the results in Table 4 have been normalized to the pH of water.

[0188] Table 4: Normalized pH and RI values ​​of selected formulations

[0189]

[0190] *pH 样品 / pH 水

[0191] As shown in Table 4, the dissolution of compound GHF-201 does not affect the pH. As for the refractive index (RI), it shows a linear behavior with the concentration of compound GHF-201, with similar slopes of about 0.001 (1%) for all systems. It is worth noting that RI is affected by the viscosity of the system, so when the viscosity increases, RI also increases. In this case, the viscosity also increases with the increase in the content of compound GHF-201, so the trend of RI with the concentration of compound GHF-201 is due to viscosity. Interestingly, when RI is normalized by viscosity, the obtained quotient decreases with the concentration of compound GHF-201 (see Figure 4A-4B Without wishing to be bound by theory, these results may be attributed to the interface of the nanostructure, which becomes more "saturated" by the compound GHF-201, thus affecting the refraction of light.

[0192] The mobility of the system components was also measured by PGSE-NMR. Tables 5-1 to 5-3 present the diffusion coefficients (×10 -11 ). The measurements were performed under a gradient / pulse program with: δ = 10.0 ms, Δ = 27.0, gmax = 44.79. The D values ​​for surfactants and oils were the average values ​​calculated for the two surfactants and MCT in each formulation.

[0193] Table 5-1: Diffusion of 8CS-RG components in concentrated and diluted form with and without compound GHF-201 Coefficient (×10 -11 )

[0194]

[0195] Table 5-2: Diffusion of LDS-C components in concentrated and diluted form with and without compound GHF-201 Coefficient (×10 -11 )

[0196]

[0197]

[0198] Table 5-3: LDS-600 system components with and without compound GHF-201 in concentrated and diluted form The diffusion coefficient (×10 -11 )

[0199]

[0200] Tables 5-1, 5-2, and 5-3 summarize the diffusion coefficients (D) of surfactants and oils (MCT) and the respective diffusion coefficients of PG, EtOH, compound GHF-201, and water. For LDS-600, the diffusion coefficient of PEG 400 was also calculated. Overall, in all test formulations, a trend of a significant decrease in the diffusion rate of all test components with the change in the concentration of compound GHF-201 in the form of concentrate was observed, indicating that the gradual loading of compound GHF-201 always resulted in a close interaction between all components. It is worth noting that when the sample is diluted, the diffusion rate of compound GHF-201 decreases, while the mobility of surfactants increases in all test systems. This indicates that compound GHF-201 plays an important role as a structural builder in the system, which helps to stabilize nanodroplets in the presence of water. This also supports the observations of DLS measurements that the droplets gradually shrink as the content of compound GHF-201 increases. On the other hand, the mobility of solvents and co-surfactants in the dilution system remains unchanged, indicating that the role of these components is to stabilize the concentrate rather than the dilution system.

[0201] Long-term physical stability

[0202] To determine the long-term stability of the formulation, use The results were quickly measured by analytical centrifugation. Figures 5A-5L as shown in . The analysis is able to predict the shelf life of a formulation at its original concentration, even in the presence of slow destabilization processes such as sedimentation, flocculation, coalescence, and fractionation. During the measurement, parallel light illuminates the entire sample cell in the centrifugal field; the transmitted light is detected by sensors arranged linearly along the total length of the sample cell. Local changes in particles or droplets are detected due to changes in light transmittance over time. The results are presented in the form of a graph that plots the percentage of transmitted light (transmittance %) as a function of local position (mm), revealing the corresponding curve of transmittance over time.

[0203] Changes in transmittance indicate the stability of the formulation - when the transmittance curve remains constant, the sample is considered physically stable and its shelf life can be inferred based on the measurement conditions.

[0204] like Figures 5A-5L As shown in , in all transmittance curves, the lines overlap, indicating that no change in transmittance was observed and all formulations were physically stable and, based on this analysis, expected to be stable for at least 2 years under storage conditions.

[0205] The physical stability of formulation LDS-C with 7.15 wt% of compound GHF-201 at 25°C and 40°C for 12 months was also evaluated. The stability test results are listed in Tables 6-1 and 6-2. The visual appearance of the samples is shown in Tables 6-1 and 6-2. Figures 6A-6C As shown in , and the DLS analysis results are shown in Figures 7A-7C as shown in .

[0206] Table 6-1: Physical stability test results, 25℃, 12 months

[0207]

[0208] Table 6-2: Physical stability test results, 40℃, 12 months

[0209]

[0210]

[0211] It is evident from the test results that the formulation is physically stable for at least 12 months at the storage temperature with no evidence of significant changes in physical properties.

[0212] Organizational infiltration

[0213] Figures 8A-8B The unformulated form ( Fig. 8A ) and formulated into LDS-C preparations ( Figure 8B ) in APBD model mice. For pharmacokinetic analysis, 100 μl of serum and brain, kidney, hind limb quadriceps, heart, liver, and spleen tissues were collected, homogenized according to established guidelines, and extracted with acetonitrile (Kapetanovic et al., 2006). A calibration curve was prepared using 0, 1, 10, 100, and 1,000 ng / ml GHF-201 in 1 mg / ml 4-tert-butyl-2-(4H-1,2,4-triazol-4-yl)phenol (ChemBridge) solution as an internal standard (IS). Tissue samples were then dissolved in 1 mg / ml IS solution and spiked with 0-1,000 ng / ml GHF-201 to generate a standard curve from which tissue levels of GHF201 were determined. Samples were analyzed by LC-MS / MS Sciex Triple Quad TM 5500 mass spectrometer.

[0214] Gbeys / ys mice injected subcutaneously with 250 mg / kg GHF-201 were sacrificed at 30, 60, 90 and 210 min after injection, and the indicated tissues were removed and 100 uL of serum was drawn. The graph shows the mean (± SEM) of GHF201 levels in different tissues determined by LC-MS / MS. Results were obtained from n=3 mice at each time point.

[0215] The distribution and kinetic parameters of GHF-201 in different tissues were evaluated. Fig. 8A and Figure 8B Comparisons between the results show that the bioavailability of GHF-201 in the orally administered formulation according to the present disclosure is increased at least three-fold in all tissues compared to GHF-201 in subcutaneous injection solution.

[0216] Pharmacokinetic testing

[0217] Adult polyglucose body disease (APBD) is a rare neurodegenerative disorder that most commonly affects adults of Ashkenazi Jewish descent. APBD is characterized by a partial loss of glycogen branching enzyme 1 (GBE1) activity, most commonly due to a mutational replacement of tyrosine 329 with a serine residue (Y329S). This loss-of-function mutation in GBE1 results in the production of poorly constructed glycogen aggregates called "polyglucose bodies" (PBs, branched amylopectin-like polysaccharides with few branch points). Because PBs are insoluble in solution and aggregates, they cannot be digested by the glycogen digestive enzyme, glycogen phosphorylase. The accumulation of aggregates often leads to liver failure and death in children (Andresen's disease; glycogen storage disease type IV; GSDIV). Milder mutations in GBE1 (such as p.Y329S) result in smaller PBs that do not interfere with hepatocytes and most other cell types and only accumulate on the sides of cells. However, in neurons and glial cells, these PBs can block the tight borders of axons over time and cause the progressive axonopathy disease APBD, which is debilitating and fatal, and is often misdiagnosed as amyotrophic lateral sclerosis or multiple sclerosis. The PB aggregates in APBD may cause different phenotypic changes, such as neurogenic bladder, partial motor dysfunction of the limbs, lower body sensory dysfunction, and in some cases also cause cognitive impairment. The feature of the progressive stage of the disease is difficulty walking, impaired balance, progressive weakness, and may even cause death. At present, there is no standard care for this disease.

[0218] Compound GHF-201 was found to reduce polyglucose in skin fibroblasts derived from APBD patients. Through computer simulation, compound GHF-201 met strict absorption, distribution, metabolism, excretion and toxicity (ADMET) criteria and was found to be safe for mice (14-day study). From a pharmacokinetic point of view, after subcutaneous injection into mice, compound GHF-201 had a long residence time and duration in the liver (>3h), moderate levels and duration in the brain and heart (1h), and negligible distribution in muscle. This pharmacokinetic profile matches the histopathological effects of compound GHF-201 on the corresponding tissues, in which polyglucose was least reduced in the liver, moderately reduced in the brain and heart, and unaffected in the muscle. Continuous treatment of APBD mice with compound GHF-201 two months before the expected onset of the disease significantly increased survival and improved motor and reflex parameters. The improvement in stretch reflex is particularly important because the patient-related symptoms are pyramidal tetraplegia or upper motor neuron signs, which is one of the main neurological defects in APBD patients.

[0219] Compound GHF-201 was administered to APBD modeling mice in a 5% DMSO solution, intravenously injected twice a week at a dose of 250 mg / kg, equivalent to a daily dose of 70 mg / kg. Due to the relatively long treatment period (6 months), daily injections were avoided, which may cause excessive scarring. After positive results in the mouse model, 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 characteristics were determined.

[0220] Blood was drawn for three consecutive days according to the following schedule - before and 0.5h, 1h, 1.5h, 2h and 4h after dosing. Compound GHF-201 was administered as formulation LDS-C (7.15% Compound GHF-201). The active ingredient dose was 170 mg on the first day, 255 mg on the second day, and 340 mg on the third day.

[0221] After blood was drawn, plasma was separated by centrifugation at 2000g for 10min (up to 1h after blood was drawn) at 4°C. The level of compound GHF-201 in plasma (supernatant) samples was determined by LC-mass spectrometry (LC-MS / MS) using a SciexTriple Quad TM 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 drawn using 0.1, 10, 100 and 1000 ng / ml compound GHF-201 in 1 mg / ml 4-tert-butyl-2-(4H-1,2,4-triazole-4-yl)phenol (ChemBridge) solution as an internal standard (IS). The sample was dissolved in a 1 mg / ml IS solution and spiked with 0-1000 ng / ml of compound GHF-201 to generate a standard curve from which the plasma level of compound GHF-201 was determined.

[0222] Figures 9A-9C The pharmacokinetic curve of patient 1 is compared with the mouse model curve. Figures 10A-10C The comparative pharmacokinetic profile of Patient 2 is shown in .

[0223] According to the FDA human equivalent dose calculation, the mouse dose can be converted to the human dose by dividing it by 12. The maximum C in mice is 4,700 ng / mL ( Fig. 9B , 10B ). 4,700 ng / mL divided by 12 gives 392 ng / mL. Patient 1's Cmax was 894 ng / mL ( Fig.9A ), and 883 ng / mL for patient 2 ( Fig. 10A ). The doses administered were found to produce a significant therapeutic effect in mice. Therefore, it was hypothesized that human equivalent doses or higher in patients would be at least as effective as the equivalent doses in mice. Another hypothesis was that the relationship between Cmax and bioactivity observed after administration of therapeutic doses would be the same as that of the doses administered, and that these results would predict therapeutic efficacy in patients with APBD.

[0224] Additionally, it was observed that administration of the LDS-C formulation compound delayed the time to Cmax in patients, indicating a "sustained release" delivery. The time to Cmax increased with increasing doses. Another interesting observation was that the pre-dose levels of compound GHF-201 in plasma increased daily ( Fig. 9C , 10C ), indicating that there is still a residual dose in the patient's plasma before the next dose is given the next day, and these doses will accumulate.

[0225] Importantly, the area under the curve (AUC) correlated with the level of the administered compound GHF-201 ( Figures 11A-11B ). This observation suggests that the body's exposure to the drug is positively correlated with the level of drug administered. AUC is calculated as the sum of trapezoids rather than using integration and extrapolation to infinite time.

[0226] Fig. 12A The muscle strength level of patient 1 (before treatment and after 10 months of treatment) is shown ( Fig. 12A ), performed according to the method described by Kleyweg et al., 1991. It can be seen that the treatment resulted in a significant increase in muscle strength in all measured muscles.

[0227] exist Figures 12B-12C Patient 1 ( Fig. 12B ) and patient 2 ( Fig. 12C NFL levels were obtained by analyzing patient plasma samples using a Simoa machine with the NF-Light v2 Advantage HD-X kit for the determination of human light chain neurofilament protein.

[0228] A reduction in neurofilament light chain levels indicates that neurodegeneration has improved. Neurofilament light chain is considered to be a systemic biomarker of the extent of neurodegeneration in several neurodegenerative diseases such as ALS and AD. Administration of GHF-201 formulated in a formulation according to the present disclosure showed a significant reduction in neurofilament light chain, indicating that the extent of neurodegeneration has improved.

[0229] Other Exemplary Formulations

[0230] Prepare a blank formulation by weighing all components according to Table 7 and mixing at 40-60 °C.

[0231] The compound of formula (II) (herein referred to as compound GHF-205) is used in other formulations:

[0232]

[0233] Table 7: Exemplary Formulation (without Compound GHF-205)

[0234]

[0235]

[0236] Compound GHF-205 was dissolved into the formulation at different concentrations (2.5 and 5.0 wt%). The formulations were tested for visual appearance, refractive index, and droplet size, as detailed in Table 8.

[0237] Table 8: Physical characteristics of GHF-205 formulations

[0238]

[0239] SD-NMR characterization was also performed on LDS-C(205) loaded with 2.5 and 5 wt% of GHF-205 in the form of concentrates, as detailed in Table 9.

[0240] Table 9: Diffusion coefficients of components in formulation LDS-C (205) (in ×10 -11 m 2 / s presents)

[0241]

[0242] *The diffusion rate of MCT is no different from that of surfactants

[0243] Overall, it can be seen that the co-surfactant in the concentrated system diffuses faster (is more mobile) than the surfactant. This suggests that while the hydrophilic surfactant and lipophilic component form the main building blocks of the structural interface in the concentrate form, upon dilution, the lipophilic component is no longer required to stabilize the interface (due to the higher mobility of the lipophilic component in the diluted form compared to the concentrate form). Therefore, in the concentrate form, the lipophilic component is located at the interface and is essential for building the droplet structure; upon dilution, the lipophilic component moves away from the interface and is located closer to the external phase. This suggests that upon oral ingestion, structural changes will allow GHF-205 to migrate out of the formulation.

Claims

1. A pharmaceutical preparation for oral delivery of a compound of formula (I) or a pharmaceutically acceptable salt, isomer or tautomer thereof, represents a single bond or a double bond, n and m are integers, each independently 1, 2 or 3; R and R 1 each independently is hydrogen or absent; R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 and R 9 are each hydrogen or are each independently selected from alkyl, cycloalkyl, alkoxy, hydroxy, thiohydroxy, thioalkoxy, aryloxy, thioaryloxy, amino, nitro, halogen, trihalomethyl, cyano, amide, carboxyl, sulfonyl, sulfonyloxy, sulfinyl and sulfonamide, each of which is 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 does not exist, and when Y is S, then R 5 does not exist; in, The preparation 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 between 0 wt % and 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.

2. The preparation according to claim 1, wherein The compound of formula (I) is a compound of formula (I') or a pharmaceutically acceptable salt, isomer or tautomer thereof, represents a single bond or a double bond, and wherein n, m, R, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 and R 8 Each of which is defined in claim 1.

3. The preparation according to claim 1 or 2, 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 preparation 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. The preparation according to any one of claims 1 to 4, which is free of water.

6. The preparation according to any one of claims 1 to 5, wherein The at least one hydrophilic surfactant is selected from the group consisting of 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. The preparation according to claim 6, wherein The formulation comprises at least one first hydrophilic surfactant selected from ethoxylated castor oil and hydrogenated derivatives thereof, and at least one second hydrophilic surfactant selected from polysorbates and ethoxylated monoglycerides.

8. The preparation according to any one of claims 1 to 7, wherein The weight ratio between total hydrophilic surfactant and total co-surfactant ranges from about 3:1 to 1:

3.

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

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

2.

12. The preparation according to 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. The preparation according to any one of claims 1 to 12, wherein The at least one oil is present in the formulation at a concentration of no more than 4 wt%.

14. A formulation according to 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 preparation according to any one of claims 2 to 14, wherein n and m are 1.

16. The preparation according to any one of claims 2 to 15, wherein R 2 , R 7 and R 8 Each is a methyl group.

17. The preparation according to any one of claims 1 to 16, wherein The compound is at least one compound of formula (IA) or (IB):

18. A pharmaceutical formulation for oral delivery of a compound of formula (II) or a pharmaceutically acceptable salt, isomer or tautomer thereof, in, The preparation 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 between 0 wt % and 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.

19. The pharmaceutical formulation according to claim 18, comprising said at least one hydrophilic surfactant in a total amount ranging between about 20 wt% and 50 wt%.

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

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

23. The preparation according to claim 22, wherein The nanodroplets have an average droplet size ranging from about 5 nm to 50 nm.

24. The preparation according to claim 22 or 23, wherein The at least one aqueous diluent is selected from the group consisting of water, water for injection, saline, dextrose solution and buffer.

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

26. The preparation according to claim 25, wherein The nanodroplets have an average droplet size ranging from about 5 nm to 50 nm.

27. The preparation according to claim 25 or 26, wherein The at least one aqueous diluent is selected from the group consisting of water, water for injection, saline, dextrose solution and buffer.

28. Use of a formulation according to any one of claims 1 to 21 or a preparation according to any one of claims 22 to 27 for the treatment of glycogen storage disease (GSD).

29. The preparation or preparation for use according to 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 VIII, GSD Type IX, GSD Type X, GSD Type XI, GSD Type XII, GSD Type XIII, GSD Type XIV, or GSD Type XV.

30. The preparation or formulation for use according to claim 28, wherein: GSD is adult polyglucosidase disorder (APBD), Anderson disease, Forbes disease, or Danon disease.

31. Use of a formulation according to any one of claims 1 to 21 or a preparation according to any one of claims 22 to 27 for treating a disease or disorder associated with lysosomal storage.

32. The preparation or preparation for use according to claim 31, wherein The disease or disorder is selected from Gaucher disease, Fabry disease, Tay-Sachs disease, mucopolysaccharidosis (MPS) disorders, aspartylglucosaminuria, GMI-gangliosidosis, Krabbe disease (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 type 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. Use of the formulation according to any one of claims 1 to 21 or the preparation according to any one of claims 22 to 27 for treating a disease or disorder associated with dysregulation of autophagy.

34. The preparation or preparation for use according to claim 33, wherein: The disease or disorder is selected from Alzheimer's disease and a cancer associated with reduced autophagic activity.

35. A method of treating glycogen storage disease (GSD) 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.

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. A method for treating a disease or condition associated with autophagy dysregulation, 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.

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