Treatment of protein aggregation myopathic and neurodegenerative diseases by parenteral administration of trehalose

Abstract

To provide a pharmaceutical formulation comprising trehalose for use in treating myopathies, neurodegenerative diseases, or tauopathies associated with abnormal protein aggregation.SOLUTION: A pharmaceutical formulation comprising a therapeutically effective amount of trehalose is provided, wherein the pharmaceutical formulation is for use in a method for treating a disease associated with abnormal protein aggregation and / or inclusion bodies formation in myocytes, neurons, or extracellular compartments, or for alleviating at least one symptom associated therewith in a human subject in need thereof, the method comprising intravenously administering to the subject a pharmaceutical formulation comprising 0.1% (w / v) to 50% (w / v) of trehalose. An injectable aqueous pharmaceutical formulation comprising a therapeutically effective amount of trehalose is also provided.SELECTED DRAWING: None

Description

Technical Field

[0001] The subject matter of the present disclosure relates to the use of trehalose in the treatment of myopathies, neurodegenerative diseases, or tauopathies associated with abnormal protein aggregation.

Background Art

[0002] References considered relevant as background to the subject matter of the present disclosure are listed below. TIFF2025087858000001.tif161160 TIFF2025087858000002.tif162158The identification of the above references in this specification is not intended to imply that they have any relevance to the patentability of the subject matter of the present disclosure.

[0003] (Background) Several groups of diseases are known to result from trinucleotide repeat mutations. These diseases are characterized by abnormal stretches of amino acids in specific proteins encoded by the mutated genes. The mutant proteins aggregate in cells, giving rise to typical cytotoxic cell inclusions.

[0004] Diseases identified as protein-codon repeat diseases include increases in homopolymeric stretches of amino acids, particularly polyglutamine (polyQ) or polyalanine (polyA). At least eight neurodegenerative diseases are associated with an increase in polyglutamine, including Huntington's disease (HD), spinal and bulbar muscular atrophy (SBMA), dentatorubral-pallidoluysian atrophy (DRPLA), and various forms of spinocerebellar ataxia (SCA). Increases in polyalanine are associated with various rare and severe congenital anomalies and oculopharyngeal muscular dystrophy (OPMD). Additional pathologies associated with increases in polyalanine would include dystrophic diseases.

[0005] Some such diseases include oculopharyngeal muscular dystrophy (OPMD); a rare hereditary myopathy characterized by ptosis, severe dysphagia, and proximal limb muscle weakness. Its estimated prevalence is 1:100,000, and the largest reported populations were French-Canadian families in Canada and the United States (prevalence 1:1000), Bukhara Jews in Israel (prevalence 1:600), and Latinos in New Mexico, Arizona, and California [1-4]. OPMD is inherited as an autosomal dominant trait with almost complete penetrance in most cases. The disease has an equal morbidity between genders. The gene associated with the disease encodes polyadenylate-binding protein nuclear 1 protein (PABPN 1); a nuclear protein involved in pre-mRNA polyadenylation, transcriptional regulation, and mRNA nuclear-cytoplasmic transport. Mutations that cause OPMD result in the production of abnormal poly(A) PABPN 1.

[0006] The disease is most often diagnosed in the fourth and fifth decades of life and progresses throughout the patient's lifetime. By the age of 70, the majority of patients develop all or some of the following symptoms: severe dysphagia, ptosis, lingual atrophy, and weakness, proximal lower and upper limb weakness, dysarthria, upward gaze limitation, and facial muscle weakness. Ptosis becomes more prominent, so patients adopt a head tilt, upward gaze in the "astronomer's position," and further worsening of dysphagia. Dysphagia begins with difficulty swallowing solid foods and progresses to liquids as well. Dysphagia becomes more severe, so patients develop malnutrition, cachexia, dehydration, and recurrent aspiration pneumonia. OPMD does not appear to shorten life expectancy but is associated with severe debilitation and a decreased quality of life.

[0007] There is no drug therapy for OPMD, and there is no possibility of cure. The current treatment strategy is limited to surgical intervention aimed at alleviating ptosis. Repeated cricopharyngeal muscle dilation is often used to relieve dysphagia. Upper esophageal sphincter resection is also used. These methods only provide temporary relief, do not affect the progression of the disease, and ultimately result in severe dysphagia, an increased risk of aspiration pneumonia due to recurrent aspiration, and severe weight loss, which are the most common causes of mortality in OPMD patients.

[0008] Therefore, there is an urgent need for compositions and treatment methods for alleviating the signs and symptoms of oculopharyngeal muscular dystrophy. Although there is controversy about the specific role of protein aggregates in the basic mechanism of cell degeneration, the widely accepted view is that protein aggregates, the aggregation process, and / or early oligomeric species are toxic and extremely important for degenerative lesions. Important evidence has been provided by studies showing that when detected early, the neurodegenerative process can be stopped and even reversed [5]. Furthermore, newer studies have shown that the turnover of many causative proteins is rapid rather than slow [6, 7], and that inclusion body formation not only causes neurotransmitter abnormalities and, most importantly, behavioral abnormalities but is also reversible.

[0009] One of the novel therapeutic strategies is, for example, to enhance the natural cellular defense mechanism against misfolded aggregated proteins by using molecular chaperones that promote re-holding of normal holding and abnormal conformations to return to the natural state [8]. Agents such as geldanamycin can regulate and enhance chaperone levels [9]. However, geldanamycin is quite toxic and does not penetrate well through the blood-brain barrier. Geldanamycin can also stimulate proteasome activity, but this approach may risk changing the metabolic turnover of molecules that are normally regulated by proteasome degradation. Furthermore, oral administration of trehalose, a disaccharide with known protein-stabilizing effects, has been shown to reduce polyglutamine aggregates, improve motor dysfunction, and extend survival in an HD mouse model

[10] . More recently, these findings have been replicated with oral administration of trehalose in a mouse model of OPMD

[11] .

[0010] Trehalose, a glucose disaccharide (α-G-glucopyranosyl α-D-glucopyranoside), is found in many plants, fungi, bacteria, insects, and other invertebrates and serves as a natural excipient. Due to its unique physical and biochemical properties demonstrated by its ability to maintain and preserve numerous biomolecules, trehalose has found uses in various foods, cosmetics, and especially therapeutic agents. Trehalose is an approved ingredient in all major markets designated as a GRAS food ingredient by the FDA

[12] . Currently, a clinical trial is being conducted in the United States to test the effect of a large oral dose of trehalose in the prevention of arterial aging. In this trial, researchers are comparing the effects of trehalose or maltose, or a combination of the two drugs, on biomarkers related to arterial aging

[17] .

Summary of the Invention

Problems to be Solved by the Invention

[0011] In one aspect thereof, the subject matter of the present disclosure is a method for treating or alleviating a disease associated with abnormal protein aggregation and / or inclusion body formation in muscle cells, neurons, and other cells, or the extracellular compartment of a human subject in need thereof, or at least one associated symptom thereof, the method comprising parenterally administering to the human subject a therapeutically effective amount of trehalose, or a pharmaceutical formulation comprising a therapeutically effective amount of trehalose. The present disclosure includes the following aspects; [Item 1] A method for treating or alleviating a disease associated with abnormal protein aggregation and / or inclusion body formation in muscle cells, neurons, and other cells, or the extracellular compartment of a human subject in need thereof, or at least one associated symptom thereof, the method comprising parenterally administering to the subject a therapeutically effective amount of trehalose or a pharmaceutical formulation comprising a therapeutically effective amount of trehalose; [Item 2] The method according to Item 1, wherein the disease is any one of polyalanine aggregation disorder, polyglutamine aggregation disorder, and tauopathy; [Item 3] The method according to Item 1 or 2, wherein the disease is any one of oculopharyngeal muscular dystrophy (OPMD), spinocerebellar ataxia (SCA), Friedreich's ataxia, spinal and bulbar muscular atrophy (SBMA), Huntington's disease, Parkinson's disease, Alzheimer's disease, and amyotrophic lateral sclerosis (ALS), dentatorubral-pallidoluysian atrophy (DRPLA), Pick's disease, corticobasal degeneration (CBD), progressive supranuclear palsy (PSP), and frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17); [Item 4] The method according to any one of Items 1 to 3, wherein the pharmaceutical formulation is an injectable solution for parenteral administration; [Item 5] The method according to any one of Items 1 to 4, wherein the pharmaceutical formulation contains trehalose as the sole active ingredient and optionally further contains at least one pharmaceutically acceptable additive, carrier, excipient, or diluent; [Item 6] The method according to any of the preceding paragraphs, wherein the trehalose concentration in the preparation is from about 0.1% (w / v) to about 50% (w / v); [Paragraph 7] The method according to any of the preceding paragraphs, wherein the concentration of trehalose in the preparation is about 10% (w / v); [Paragraph 8] The method according to any of the preceding paragraphs, wherein the pharmaceutical preparation containing trehalose has an osmolality of from about 280 to about 330 mOsm / Kg; [Paragraph 9] The method according to any of paragraphs 1 to 8, wherein the administration is any of intravenous, intramuscular, and intraperitoneal administrations; [Paragraph 10] The method according to any of the preceding paragraphs, wherein the pharmaceutical preparation contains less than 0.74 endotoxin units / ml solution; [Paragraph 11] The method according to any of the preceding paragraphs, wherein the therapeutically effective amount of trehalose is from about 1 g to about 100 g per single injection and is not more than about 1 g / kg of the subject body weight per day; [Paragraph 12] The method according to any of paragraphs 1 to 11, wherein the therapeutically effective amount of trehalose or the pharmaceutical preparation containing the same is administered chronically; [Paragraph 13] The method according to any of paragraphs 1 to 12, wherein the therapeutically effective amount of trehalose or the pharmaceutical preparation containing the same is administered at a frequency of once a day to once a month; [Paragraph 14] The method according to paragraph 13, wherein the therapeutically effective amount of trehalose or the trehalose contained in the pharmaceutical preparation is administered once a day at about 10 mg / kg / day to about 1 g / kg / day of trehalose; [Paragraph 15] The method according to any of paragraphs 1 to 12, wherein the therapeutically effective amount of trehalose or the trehalose contained in the pharmaceutical preparation is administered once a week by single injection; [Paragraph 16] An effective amount of trehalose or a pharmaceutical composition containing the same is intravenously administered in a single dose of about 5 to about 35 g of trehalose, and the administration can be once a day, every other day, twice a week, once a week, once every two weeks, once every three weeks, or once a month, according to the method described in item 13 or 15; [Item 17] The method according to item 13 or 15, wherein an effective amount of trehalose or trehalose contained in the pharmaceutical preparation is administered in a single dose of 5, 8, 15, 30, 40, or 50 g of trehalose; [Item 18] The method according to any one of items 1 to 17, wherein the pharmaceutical preparation is an injectable solution and the administration rate is such that the maximum endotoxin level is less than 5 endotoxin units / kg body weight / hour; [Item 19] The method according to any one of items 1 to 18, wherein the administration of the effective amount of trehalose contained in the pharmaceutical preparation adapted for intravenous administration is completed within about 75 to about 120 minutes, specifically less than 90 minutes; [Item 20] The method according to any one of items 12 to 19, wherein the administration includes a dosing schedule of trehalose or a pharmaceutical preparation containing the same at an equal dose, or a gradually increasing dose, or a gradually decreasing dose; [Item 21] The method according to any one of items 1 to 11, wherein the effective amount of trehalose or a pharmaceutical preparation containing the same is administered regularly; [Item 22] A method for treating or alleviating a disease associated with abnormal protein aggregation and / or inclusion body formation in muscle cells, neurons, and other cells, or the extracellular compartment of a human subject in need thereof, or at least one related symptom thereof, the method comprising parenterally administering to the subject a therapeutically effective amount of trehalose or a pharmaceutical preparation containing the same. A trehalose or a pharmaceutical preparation containing the same for use in the method; [Item 23] A trehalose or a pharmaceutical preparation containing the same for use as described in item 22, wherein the disease is any one of polyalanine aggregation disorder, polyglutamine aggregation disorder, and tauopathy; [Item 24] Trehalose or a pharmaceutical preparation containing the same for use as described in item 22 or 23, wherein the disease is any one of oculopharyngeal muscular dystrophy (OPMD), spinocerebellar ataxia (SCA), Friedrich's ataxia, spinal and bulbar muscular atrophy (SBMA), Huntington's disease, Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), dentatorubral-pallidoluysian atrophy (DRPLA), Pick's disease, corticobasal degeneration (CBD), progressive supranuclear palsy (PSP), and frontotemporal dementia and Parkinsonism linked to chromosome 17 (FTDP-17); [Item 25] Trehalose or a pharmaceutical preparation containing the same for use as described in any one of items 22 to 24, wherein the pharmaceutical preparation is an injectable solution for parenteral administration; [Item 26] Trehalose or a pharmaceutical preparation containing the same for use as described in any one of items 22 to 25, wherein the pharmaceutical preparation contains trehalose as the sole active ingredient and optionally further contains at least one pharmaceutically acceptable additive, carrier, excipient, or diluent; [Item 27] Trehalose or a pharmaceutical preparation containing the same for use as described in any one of items 22 to 26, wherein the trehalose concentration in the preparation is from about 0.1% (w / v) to about 50% (w / v); [Item 28] Trehalose or a pharmaceutical preparation containing the same for use as described in any one of items 22 to 27, wherein the trehalose concentration in the preparation is about 10% (w / v); [Item 29] Trehalose or a pharmaceutical preparation containing the same for use as described in any one of items 22 to 28, wherein the pharmaceutical preparation containing trehalose has an osmolality of about 280 to 330 mOsm / Kg; [Item 30] Trehalose or a pharmaceutical preparation containing the same for use as described in any one of items 22 to 29, wherein the administration is any one of intravenous, intramuscular, and intraperitoneal administrations; [Item 31] Trehalose or a pharmaceutical preparation containing the same for use as described in any of paragraphs 22 to 30, wherein the pharmaceutical preparation contains less than 0.74 endotoxin units / ml solution; [Paragraph 32] Trehalose or a pharmaceutical preparation containing the same for use as described in any of paragraphs 22 to 31, wherein the therapeutically effective amount of trehalose is about 1 g to about 100 g per single injection and is less than about 1 g / Kg of the subject body weight per day; [Paragraph 33] Trehalose or a pharmaceutical preparation containing the same for use as described in any of paragraphs 22 to 32, wherein the therapeutically effective amount of trehalose or the pharmaceutical preparation containing the same is administered chronically; [Paragraph 34] Trehalose or a pharmaceutical preparation containing the same for use as described in any of paragraphs 22 to 33, wherein the therapeutically effective amount of trehalose or the pharmaceutical preparation containing the same is administered at a frequency of once a day to once a month; [Paragraph 35] Trehalose or a pharmaceutical preparation containing the same for use as described in paragraph 34, wherein the therapeutically effective amount of trehalose or the trehalose contained in the pharmaceutical preparation is administered once a day at a trehalose amount of about 10 mg / kg / day to about 1 g / kg / day; [Paragraph 36] Trehalose or a pharmaceutical preparation containing the same for use as described in any of paragraphs 22 to 33, wherein the therapeutically effective amount of trehalose or the trehalose contained in the pharmaceutical preparation is administered once a week by single injection; [Paragraph 37] Trehalose or a pharmaceutical preparation containing the same for use as described in paragraph 34 or 36, wherein the therapeutically effective amount of trehalose or the trehalose contained in the pharmaceutical preparation is administered intravenously once a day, every other day, twice a week, once a week, once every two weeks, once every three weeks, or once a month, at a single dose of about 5 to about 35 g of trehalose; [Paragraph 38] Trehalose in a therapeutically effective amount or a pharmaceutical preparation containing trehalose therein for use as described in item 34 or 36 by administering a single dose of 5, 8, 15, 30, 40, or 50 g of trehalose; [Item 39] Trehalose in a therapeutically effective amount or a pharmaceutical preparation containing the same for use as described in any of items 22 to 38, wherein the pharmaceutical preparation is an injectable solution and the administration rate is such that the maximum endotoxin level is less than 5 endotoxin units / kg body weight / hour; [Item 40] Trehalose in a therapeutically effective amount or a pharmaceutical preparation containing the same for use as described in any of items 22 to 39, wherein the pharmaceutical preparation is suitable for intravenous administration and the administration is completed within about 75 to about 120 minutes, particularly less than 90 minutes; [Item 41] Trehalose in a therapeutically effective amount or a pharmaceutical preparation containing the same for use as described in any of items 33 to 40, wherein the administration includes a dosing schedule of trehalose or a pharmaceutical preparation containing the same at an equal dose, or a gradually increasing dose, or a gradually decreasing dose; [Item 42] Trehalose in a therapeutically effective amount or a pharmaceutical preparation containing the same for use as described in any of items 22 to 32 by administering the same periodically; [Item 43] An aqueous pharmaceutical preparation having a pH of about 4.5 to 7.0, containing less than 0.74 endotoxin unit / ml, and being suitable for parenteral administration, and containing trehalose in a therapeutically effective amount as the sole active ingredient; [Item 44] The aqueous pharmaceutical preparation described in item 43, which is suitable for intravenous, intramuscular, or intraperitoneal administration; [Item 45] The aqueous pharmaceutical preparation described in item 43 or 44, optionally further containing at least one pharmaceutically acceptable additive, carrier, excipient, or diluent; [Item 46] An aqueous pharmaceutical preparation according to any one of Items 43 to 45, wherein the trehalose concentration is from about 0.1% (w / v) to about 50% (w / v); [Item 47] An aqueous pharmaceutical preparation according to any one of Items 43 to 46, wherein the trehalose concentration is about 10% (w / v); [Item 48] An aqueous pharmaceutical preparation according to any one of Items 43 to 47, having a molar osmotic concentration of about 280 to 330 mOsm / kg; [Item 49] An aqueous pharmaceutical preparation according to any one of Items 43 to 48, which is administered once a day to once a month; [Item 50] An aqueous pharmaceutical preparation according to any one of Items 43 to 47, wherein the therapeutically effective amount of trehalose is about 1 g to about 100 g per injection once a day and less than about 1 g / kg of the subject body weight per day; [Item 51] An aqueous pharmaceutical preparation according to any one of Items 43 to 50, wherein trehalose is administered once a day at a dose of about 10 mg / kg / day to about 1 g / kg / day; [Item 52] An aqueous pharmaceutical preparation according to any one of Items 43 to 51, which is administered at a frequency of once a day to once a month at a dose of about 5 to about 35 g of trehalose contained therein; [Item 53] An aqueous pharmaceutical preparation according to Item 52, for administration once a day, every other day, twice a week, once a week, once every two weeks, once every three weeks, or once a month; [Item 54] An aqueous pharmaceutical preparation according to Item 53, wherein the dose is 5, 8, 15, 30, 40, or 50 g; [Item 55] An aqueous pharmaceutical preparation according to any one of Items 43 to 54, wherein the administration rate is such that the maximum endotoxin level is less than 5 endotoxin units / kg body weight / hour; [Item 56] An aqueous pharmaceutical preparation according to any one of Items 43 to 55, wherein the administration is completed within about 75 to about 120 minutes, particularly less than 90 minutes; [Item 57] An aqueous pharmaceutical formulation according to any of paragraphs 43 to 56 for treating a disease associated with abnormal protein aggregation and / or inclusion body formation in muscle cells, neurons, and other cells, or extracellular compartments, or for alleviating symptoms or signs associated therewith in a human subject in need thereof; [Paragraph 58] The aqueous pharmaceutical formulation according to paragraph 57, wherein the disease is any of oculopharyngeal muscular dystrophy (OPMD), spinocerebellar ataxia (SCA), Friedreich's ataxia, spinal bulbar muscular atrophy (SBMA), Huntington's disease, Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), dentatorubral-pallidoluysian atrophy (DRPLA), Pick's disease, corticobasal degeneration (CBD), progressive supranuclear palsy (PSP), and frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17); or [Paragraph 59] A kit comprising: (a) pharmaceutically acceptable trehalose or an active derivative thereof; (b) at least one pharmaceutically acceptable additive, carrier, excipient, and diluent; (c) means for producing an injectable aqueous solution of the trehalose by mixing the trehalose with at least one of the additive, carrier, excipient, and diluent; (d) means for parenterally administering the injectable solution to a patient in need thereof; (e) instructions for use.

Means for Solving the Problem

[0012] In the foregoing and other aspects, the disease of the subject matter of the present disclosure is any one of polyalanine aggregation disorder, polyglutamine aggregation disorder, and tauopathy, such as oculopharyngeal muscular dystrophy (OPMD), spinocerebellar ataxia (SCA), Friedreich's ataxia, spinal bulbar muscular atrophy (SBMA), Huntington's disease, Parkinson's disease, Alzheimer's disease, and amyotrophic lateral sclerosis (ALS), dentatorubral-pallidoluysian atrophy (DRPLA), Pick's disease, corticobasal degeneration (CBD), progressive supranuclear palsy (PSP), and any one of frontotemporal dementia and Parkinsonism linked to chromosome 17 (FTDP-17). In the foregoing and other aspects, the pharmaceutical preparation of the subject matter of the present disclosure is an injectable solution for parenteral administration, particularly for intravenous, intramuscular, and intraperitoneal administration.

[0013] In some aspects, the pharmaceutical preparation of the subject matter of the present disclosure contains trehalose as the only active ingredient, and optionally further contains at least one pharmaceutically acceptable additive, carrier, excipient, or diluent.

[0014] In all aspects, the concentration of trehalose in the preparation of the subject matter of the present disclosure is from about 0.1% (w / v) to about 50% (w / v), particularly about 10% (w / v). In some aspects, the pharmaceutical preparation containing trehalose of the subject matter of the present disclosure has an osmolality of about 280 to about 330 mOsm / Kg.

[0015] In the foregoing and other aspects, the pharmaceutical preparation of the subject matter of the present disclosure contains less than 0.74 endotoxin units / ml solution.

[0016] The therapeutically effective amount of trehalose of the subject matter of the present disclosure is from about 1 g (gram) to about 100 g per single injection, and is less than about 1 g / kg of the subject body weight per day.

[0017] In other embodiments, a therapeutically effective amount of trehalose or trehalose contained in the pharmaceutical formulation is administered once daily as about 10 mg / kg / day to about 1 g / kg / day of trehalose. In further embodiments, a therapeutically effective amount of trehalose or trehalose contained in the pharmaceutical formulation of the subject matter of the present disclosure is administered as a single dose of 5, 8, 15, 30, 40, or 50 g of trehalose.

[0018] A therapeutically effective amount of trehalose or a pharmaceutical formulation containing the same of the present disclosure can be administered chronically or periodically.

[0019] In the above and other embodiments, a therapeutically effective amount of trehalose or a pharmaceutical formulation containing the same of the present disclosure is administered at a frequency of once a day to once a month.

[0020] In a further embodiment, a therapeutically effective amount of trehalose or trehalose contained in the pharmaceutical formulation of the subject matter of the present disclosure is administered as a single injection once a week.

[0021] In a further embodiment, a pharmaceutical formulation containing a therapeutically effective amount of trehalose or trehalose of the subject matter of the present disclosure can be administered intravenously as a single dose of about 5 to about 35 g of trehalose once a day, every other day, twice a week, once a week, once every two weeks, once every three weeks, or once a month.

[0022] In the above and other embodiments, the pharmaceutical formulation of the subject matter of the present disclosure is an injectable solution, and the administration rate is such that the maximum endotoxin level is less than 5 endotoxin units / kg body weight / hour.

[0023] In some embodiments, the administration of a therapeutically effective amount of trehalose contained in the pharmaceutical formulation suitable for intravenous administration described herein is completed within about 75 to about 120 minutes, particularly less than 90 minutes.

[0024] In other embodiments, the administration of the subject matter of the present disclosure includes a dosing schedule of trehalose or a pharmaceutical formulation containing the same at an equal dose, or a gradually increasing dose, or a gradually decreasing dose.

[0025] In another aspect of that situation, the subject matter of the present disclosure is a method for treating or alleviating a disease associated with abnormal protein aggregation and / or inclusion body formation in muscle cells, neurons, and other cells, or the extracellular compartment of a human subject in need thereof, or at least one related symptom thereof, the method comprising parenterally administering to the human subject a therapeutically effective amount of trehalose or a pharmaceutical formulation containing the same. Provided is trehalose or a pharmaceutical formulation containing the same for use in such a method.

[0026] In a further aspect, the subject matter of the present disclosure is an aqueous pharmaceutical formulation containing a therapeutically effective amount of trehalose as the sole active ingredient, having a pH of about 4.5 to 7.0, containing less than 0.74 endotoxin units / ml, and provided is such a pharmaceutical formulation suitable for parenteral administration.

[0027] Furthermore, the subject matter of the present disclosure provides a kit comprising: (a) pharmaceutically acceptable trehalose or an active derivative thereof; (b) at least one pharmaceutically acceptable additive, carrier, excipient, and diluent; (c) means for producing an injectable aqueous solution of the trehalose by mixing the trehalose with at least one of the additive, carrier, excipient, and diluent; (d) means for parenterally administering the injectable solution to a patient in need thereof; (e) instructions for use.

[0028] To better understand the subject matter described herein and to illustrate the method of practicing it, embodiments are described as non-limiting examples with reference to the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0029]

Figure 1

Figure 2

DETAILED DESCRIPTION OF THE INVENTION

[0030] The subject matter of the present disclosure is based on the surprising finding that when an aqueous formulation containing trehalose is administered parenterally, for example intravenously, a therapeutically effective amount of trehalose reaches the plasma and accumulates in the muscle of the test animal, as compared to the plasma and muscle levels of trehalose obtained after oral administration of such a formulation.

[0031] Trehalose is known for its ability to maintain and preserve a number of biomolecules. Trehalose is used in various research applications and mainly serves as a lyoprotectant and is included in various commercially available therapeutic agents including Herceptin, Avastin, Lucentis, and Advate. Trehalose is widely used as an excipient / additive together with another active ingredient, but its use as the therapeutic active ingredient itself is rather exceptional.

[0032] Trehalose can inhibit the intracellular aggregation of abnormal proteins associated with neurodegenerative diseases and myopathies. A major example is the aggregation inhibitory effect of trehalose on mutant huntingtin, a polyglutamine protein that causes Huntington's disease (HD). Oral administration of trehalose to transgenic HD mice (naturally consumed 0.2% - 5% trehalose in drinking water over 5 - 9 weeks) resulted in inhibition of the formation of nuclear huntingtin aggregates in the brain and liver, and more importantly, improvement of HD-related motor symptoms.

[0033] Interestingly, particularly in neurological and muscular diseases, the therapeutic use of trehalose is severely hampered because trehalose is hydrolyzed by the endogenous enzyme trehalase at the epithelial brush border of the small intestine [13, 14], so that only a small fraction of any orally administered dose reaches the bloodstream, nerves, or muscle tissue. The present disclosure aims to introduce a new treatment regimen that achieves higher bioavailability and therapeutic efficacy in the treatment of myopathies and neurodegenerative diseases (particularly polyalanine polyglutamine), and tauopathies, disorders, and particularly OPMD, associated with abnormal protein aggregation using parenteral administration of trehalose.

[0034] Accordingly, the subject matter of the present disclosure is a method for treating or alleviating a disease associated with abnormal protein aggregation and / or inclusion body formation in muscle cells, neurons, and other cells, or extracellular compartments of a human subject in need thereof, or at least one associated symptom thereof, the method comprising parenterally administering to the human subject a therapeutically effective amount of trehalose or a pharmaceutical formulation comprising a therapeutically effective amount of trehalose.

[0035] As used herein, the term "disease associated with abnormal protein aggregation and / or inclusion body formation in muscle cells, neurons, and other cells, or extracellular compartments" refers to any disease associated with protein aggregation or protein misfolding, the common biological feature underlying these diseases being that a cascade of pathological events occurs, including the aggregation of certain peptides and proteins and the secondary aggregation of various other proteins thereby, resulting in a failure of protein homeostasis to maintain normal biological functions.

[0036] As used herein, the term "treating or alleviating" refers to achieving a therapeutic effect of improving, alleviating, or reducing the severity and / or frequency of at least one sign or symptom associated with the disease described herein, removing the sign or symptom and / or the underlying cause, preventing the occurrence of the symptom and / or its underlying cause (e.g., prophylactic treatment), improving or repairing the damage, and removing or reducing the degree of protein aggregation.

[0037] Symptoms associated with the diseases described herein include, but are not limited to, ptosis of the eyelids (a condition known as "blepharoptosis"), dysphagia (referred to as "dysphagia"), muscle fatigue, activity / movement disorders, and cognitive impairment.

[0038] In particular, when the disease to be treated is OPMD, the formulations and methods described herein are useful for treating the signs and symptoms of OPMD. Signs and symptoms of OPMD include severe dysphagia, ptosis, lingual atrophy, and weakness, proximal lower and upper limb weakness, dysarthria, upward gaze limitation, and facial muscle weakness.

[0039] As is known in the art, a number of diseases and disorders are defined as "diseases associated with abnormal protein aggregation and / or inclusion body formation in muscle cells, neurons, and other cells, or the extracellular compartment". For example, the subject matter of the present disclosure provides methods for treating or alleviating diseases associated with abnormal protein aggregation, which include, by way of example, neurodegenerative diseases, polyglutamine or polyalanine aggregation disorders, protein codon repeat disorders, myopathies, and tauopathies.

[0040] As used herein, the term "neurodegenerative disease" refers to a hereditary or sporadic condition characterized by progressive dysfunction of the nervous system, particularly disorders in this group associated with the formation of abnormal protein aggregates also known as inclusion bodies. Examples of neurodegenerative diseases associated with abnormal protein aggregation include amyotrophic lateral sclerosis (ALS), Alzheimer's disease, Parkinson's disease, and others.

[0041] As used herein, the terms "polyglutamine", "polyalanine aggregation disorder", or "protein codon repeat disorder" refer to disorders associated with the formation of intracellular polyglutamine or polyalanine aggregates, and preferably represent oculopharyngeal muscular dystrophy (OPMD), Huntington's disease (HD), spinal and bulbar muscular atrophy (SBMA), dentatorubral-pallidoluysian atrophy (DRPLA), and spinocerebellar ataxia (SCA).

[0042] As used herein, the term "myopathy" refers to a hereditary or acquired degenerative disease associated with muscle fiber atrophy, particularly in the context of the present disclosure where it represents OPMD.

[0043] As used herein, the term "tauopathy" refers to a neurodegenerative disease associated with tau lesions and the prototypical intracellular aggregation of tau microfilaments, particularly in the context of the present disclosure where it represents Pick's disease, corticobasal degeneration (CBD), progressive supranuclear palsy (PSP), and frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17).

[0044] Tauopathy is known as a disease caused by mutations that result in the misfolding of the tau microtubule-associated protein that binds to and stabilizes the microtubules of nerve cells. Tau lesions are a prominent feature of sporadic Alzheimer's disease (AD), but are also seen in various other related neurodegenerative diseases, such as Pick's disease, corticobasal degeneration (CBD), progressive supranuclear palsy (PSP), and frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17). More than 30 various hereditary mutations or nucleotide substitutions in the FTDP-17 gene on chromosome 17q21 are associated with neurodegenerative diseases in which prototypical intracellular aggregation of tau microfilaments appears. Tau mutations and tau dysfunction in similar hereditary and sporadic diseases can lead to disease through mechanisms involving both loss of function (decreased microtubule stability) and toxic gain of function (increased protofibril formation).

[0045] Accordingly, in the above and other embodiments, the disease is any one of polyalanine aggregation disorder, polyglutamine aggregation disorder, and tauopathy. In the above and other embodiments, the disease is, as described herein, oculopharyngeal muscular dystrophy (OPMD), spinocerebellar ataxia (SCA), Friedreich's ataxia, spinal bulbar muscular atrophy (SBMA), Huntington's disease, Parkinson's disease, Alzheimer's disease, and amyotrophic lateral sclerosis (ALS), dentatorubral-pallidoluysian atrophy (DRPLA), Pick's disease, corticobasal degeneration (CBD), progressive supranuclear palsy (PSP), and frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17). In some embodiments, the disease is oculopharyngeal muscular dystrophy (OPMD) as described herein.

[0046] As described above, OPMD results from PABPN1 misfolding and a hereditary abnormal increase in polyalanine in the poly(A)-binding protein nuclear 1 protein (PABPN1) that leads to the formation of tubular-filamentous inclusions / aggregates in the nuclei of affected muscle cells. Oral administration of trehalose to transgenic OPMD mice (spontaneously ingesting 2% trehalose in drinking water for 4 - 6 months) has been shown to significantly reduce the toxicity and aggregate formation of mutant PABPN1 in muscle cells. Overall, oral trehalose showed a delay in OPMD onset, attenuation of the disease phenotype, reduction of polyalanine protein aggregate formation, and reduction of cell death, suggesting that trehalose could be a potent anti-aggregation therapy for OPMD and other protein codon repeat disorders.

[0047] As shown in the examples, the inventors have shown that trehalose can be successfully delivered to muscle, which is the target organ in the treatment of various diseases and disorders described herein, such as but not limited to OPMD.

[0048] Huntington's disease and OPMD both have a common genetic basis in that they are caused by a hereditary trinucleotide expansion mutation of the gene responsible for the disease. The HD gene (HTT) on chromosome 4p16 contains a repetitive trinucleotide sequence (CAG)n that encodes a polyglutamine (polyQ) stretch. Normal HTT gene variants contain 7 - 35 repeats, while HD-related variants are above that range, and the degree of increase in repeats is associated with earlier onset and severity of HD symptoms (this effect is expected). An increased mutation of the similar (CAG)n repeat is associated with various ataxias belonging to the group of other polyglutamine disorders, such as SBMA, DRPLA, and SCA. Similarly, the OPMD-related gene, PABPN1, on chromosome 14q11 contains a trinucleotide repeat sequence (CGC)n that encodes polyalanine (polyA), with (CGC)6 repeats being the normal threshold, and OPMD beyond that is predicted by an increase in the number of repeats. Thus, in other aspects, the disease is Huntington's disease as described herein.

[0049] Except for drug therapies that alleviate some motor and psychiatric symptoms, it is clear that there is no drug therapy for HD, OPMD, and other related disorders, and there is a great need for therapies that directly inhibit the mechanisms underlying the disease.

[0050] In one aspect, the disease is spinocerebellar ataxia (SCA) as described herein. As is known in the art, there are many types of spinocerebellar ataxia, and the most common spinocerebellar ataxias include Friedreich's ataxia, SCA 1, 3, 8, etc. Many SCAs are included in the category of polyglutamine diseases that occur when the disease-related protein (i.e., ataxin-1, ataxin-3, etc.) contains a glutamine repeat exceeding a certain threshold.

[0051] In another aspect, the disease is spinal bulbar muscular atrophy (SBMA) as described herein. As is known in the art, the disease SBMA, also known as spinal bulbar muscular atrophy, bulbar spinal atrophy, X-linked spinal bulbar neuropathy (XBSN), X-linked spinal muscular atrophy type 1 (SMAX1), and Kennedy disease (KD), is a debilitating neurodegenerative disease that causes muscle cramps and progressive weakness due to the degeneration of motor neurons in the brainstem and spinal cord.

[0052] In a further aspect, the disease is Huntington's disease, Parkinson's disease, Alzheimer's disease, or amyotrophic lateral sclerosis (ALS) as described herein. Huntington's disease (HD) is a neurodegenerative genetic disease that affects muscle coordination and causes cognitive decline and psychiatric problems; Parkinson's disease is a degenerative disease of the central nervous system; Alzheimer's disease (AD) exhibits short-term memory loss, confusion, irritability, aggression, mood swings, speech problems, and long-term memory loss; and amyotrophic lateral sclerosis (ALS, also known as motor neuron disease or Lou Gehrig's disease) is a neurodegenerative disease characterized by muscle atrophy and rapid progressive weakness due to muscle cramps, difficulty speaking (dysarthria), difficulty swallowing (dysphagia), and difficulty breathing (respiratory distress).

[0053] As described above, the subject matter of the present disclosure is a method for treating or alleviating a disease as described herein in a human subject in need thereof, the method comprising parenterally administering to the human subject a therapeutically effective amount of trehalose or a pharmaceutical formulation comprising the same. In the above report of trehalose as a therapeutic agent for diseases associated with protein aggregation, extremely high continuous oral intake of trehalose is required to achieve the desired therapeutic effect in neural or muscle tissue.

[0054] The present disclosure is directed to a treatment regimen using trehalose administered by parenteral administration to a subject in need thereof. Thus, in one aspect, the pharmaceutical formulation described herein is an injectable solution for parenteral administration.

[0055] The term "subject in need thereof" includes, but is not limited to, subjects suffering from a disease described herein, such as HD, or OPMD, or another polyglutamine or polyalanine aggregation disorder.

[0056] A therapeutically effective amount of trehalose can be from about 10 mg to about 1,000 mg / Kg body weight / day. The trehalose can be included as an active ingredient in a pharmaceutical formulation suitable for parenteral administration. Administration can be based on a daily dosing schedule as a single dose or multiple doses by single parenteral administration or multiple doses by multiple parenteral administrations, respectively. Alternatively or additionally, administration can be made periodically, for example, every other day, three times a week, twice a week, once a week, or once a month, and the dosing frequency can be varied according to the patient's condition.

[0057] As used herein, parenteral refers to an administration route where the desired effect is systemic and the active substance (trehalose as described herein) is administered by a route other than the digestive tract, such as intravenous, intramuscular, and intraperitoneal administrations.

[0058] As noted above, trehalose is known in the art as a lyoprotectant. Thus, the safety and toxicity of trehalose have been extensively studied, and the substance has been found to be safe when administered orally and intravenously at doses considerably higher than the intended therapeutic dose.

[0059] As shown in the following examples, parenteral administration of a pharmaceutical formulation containing trehalose as the sole active ingredient resulted in relatively high plasma and muscle levels.

[0060] Thus, in one aspect, the pharmaceutical formulations described herein contain trehalose as the sole active ingredient and, optionally, further contain at least one pharmaceutically acceptable additive, carrier, excipient, or diluent.

[0061] That is, in the subject matter of the present disclosure, trehalose is the sole active substance or ingredient. Nevertheless, trehalose or a pharmaceutical formulation containing the same can be used in combination with other therapies and treatments for treating or alleviating diseases associated with abnormal protein aggregation.

[0062] As used herein, the term "trehalose" refers to the disaccharide glucose α-G-glucopyranosyl α-D-glucopyranoside. The term trehalose also refers to any active derivatives, hydrides, and salts thereof.

[0063] Trehalose is a natural disaccharide consisting of a 1,1-linkage of two D-glucose molecules. Trehalose is a non-reducing sugar that is not readily hydrolyzed by acids. Its molecular formula is C 12 H 22 O 11 and its molecular weight is 342.31 daltons. Other names used to represent the α,α form of the dimer commonly referred to as "trehalose" are α,α-trehalose, α-δ-glucopyranosyl, α-δ-glucopyranoside, mushroom sugar, mycose.

[0064] As described above, trehalose is well known for its protein stabilizing properties. Trehalose is widely used in many applications as a stabilizer for frozen foods, freeze-drying of biological systems and cells, as a stabilizer for therapeutic parenteral proteins, and as a carrier for tablets and IV solutions. Trehalose is recognized by the FDA as a GRAS (Generally Recognized as Safe) food ingredient and is described in the USP-NF (United States Pharmacopeia-National Formulary), EP (European Pharmacopoeia), and JP (Japanese Pharmacopoeia).

[0065] Like all disaccharides, trehalose is metabolized to two D-glucose molecules at the epithelial brush border. Less than 0.5% of digested trehalose is absorbed into the bloodstream and is further metabolized by trehalase in the liver and kidneys. Oral trehalose in amounts exceeding 40 - 50 g / day causes diarrhea and a feeling of fullness. Therefore, it was necessary to avoid extensive metabolism in the gastrointestinal (GI) tract to achieve therapeutic amounts of trehalose in muscle cells. Accordingly, the inventors developed an intravenous (IV) solution of trehalose.

[0066] As used herein, the term "formulation" refers to a composition comprising trehalose as an active ingredient and, optionally, further comprising additional active ingredients, such as anti-inflammatory agents well known in the art, and at least one pharmaceutically acceptable additive, carrier, excipient, or diluent. This formulation may further contain a trehalase (a glycoside hydrolase that catalyzes the conversion of trehalose to glucose in the intestine, kidneys, and liver) inhibitor, i.e., a competitive or other inhibitor of the trehalase enzyme.

[0067] Certain pharmaceutical formulations, particularly injectable solutions in which the trehalose concentration in the formulation is from about 0.1% (weight / volume) to about 50% (weight / volume), more specifically, although not limited to, about 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, or 25% (weight / volume) of trehalose as described herein, are suitable for parenteral administration.

[0068] With respect to trehalose, the term trehalose includes, but is not limited to, certain molecular species such as salts, polymorphs, esters, amides, prodrugs, adducts, conjugates, active metabolites, etc., and pharmaceutically acceptable, pharmacologically active analogs thereof.

[0069] Accordingly, the active substance trehalose can be administered in the form of the compound itself and in the form of salts, polymorphs, esters, amides, prodrugs, derivatives, etc., provided that the salts, polymorphs, esters, amides, prodrugs, or derivatives are pharmacologically suitable. Salts, esters, amides, prodrugs, and other derivatives of the active substance can be prepared using standard procedures known to those skilled in the field of synthetic organic chemistry, such as those described in J. March, Advanced Organic Chemistry: Reactions, Mechanisms and Structure, 4th Edition (New York: Wiley-Interscience, 1992). For any active substance that may exist in enantiomeric form, the active substance can be incorporated into the formulations of the present invention in racemic compound or enantiomerically pure form.

[0070] Salts are compounds that ionize in aqueous solutions and can be used, for example, to adjust the osmotic pressure of such solutions. When the active agent is present in the form of a salt, additional salts can be added to the composition, for example, to ion-exchange the active substance. Suitable salts for use in the compositions described herein are known in the art and include, for example, lithium, sodium, potassium, calcium, and magnesium salts having appropriate counterions that can be selected from, for example, chloride, bromide, iodide, carbonate, phosphate, nitrate, silicate, sulfate, phosphite, nitrite, sulfite, etc.

[0071] Buffers are compounds or solutions used to help maintain the concentration of a specimen within a desired range. For example, pharmaceutically acceptable pH buffers are used to maintain the acidity or basicity of a solution within a pharmaceutically acceptable range. Buffers for use in the compositions described herein can be any known or hereinafter described buffers.

[0072] An excipient is an inert ingredient that can be used in the compositions described herein for various reasons. A wide range of excipients are described in the literature (e.g., Rowe et al., Handbook of Pharmaceutical Excipients, McGraw Hill, 2006). Additives and diluents are well known in the art.

[0073] In certain embodiments, the pH of the formulation is from about 4.5 to 7.0, the osmolality of the formulation is from about 280 to 330 mOsm / kg, the formulation contains 1.0, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1 endotoxin units / ml or less, and the aqueous formulation is about 50%, 40%, 30%, 20%, 10%, 5% trehalose (w / v) or less.

[0074] Thus, the trehalose delivery systems of the present disclosure, such as formulations containing trehalose as the sole active ingredient, can generally incorporate into the formulation a buffer (pH adjusting additive) known in the art, a substance for adjusting its osmolality, and optionally one or more pharmaceutically acceptable carriers, excipients, and / or additives.

[0075] For the formulations described herein that are administered as aqueous or other solvent-based dosage forms (e.g., for parenteral administration), various aqueous carriers (especially water or physiological saline) can be used. Aqueous solutions include salts, buffers, etc. The carrier can be a solvent or dispersion medium suitable for parenterally administrable compositions including, for example, water, physiological saline, ethanol, polyols (such as glycerol, propylene glycol, and liquid polyethylene glycols, etc.), suitable mixtures thereof, and vegetable oils. Suitable fluidity can be maintained, for example, by using a coating such as lecithin, maintaining the required particle size in the case of a dispersion, and using surfactants. Water is an essential additive (or carrier).

[0076] Accordingly, the term "pharmaceutically acceptable carrier" as used herein includes all solvents, dispersion media, coatings, antibacterial and antifungal agents, and the like. The use of such media and substances for pharmaceutical active substances is well known in the art. The use of any conventional media or substances in pharmaceutical compositions is contemplated, except where they are incompatible with the active ingredient. The active substances are expected to be deliverable by any acceptable parenteral route in any pharmaceutically acceptable dosage form.

[0077] For parenteral administration, suitable oils such as sesame oil or peanut oil, or sterile aqueous solutions of the corresponding water-soluble salts in aqueous propylene glycol can be used. Such aqueous formulations can be buffered appropriately if necessary, and the liquid diluent is first made isotonic with sufficient physiological saline or glucose. These aqueous formulations are particularly suitable for intravenous, intramuscular, subcutaneous, and intraperitoneal injection. In this regard, all the sterile aqueous media used can be readily obtained by standard techniques well known to those skilled in the art.

[0078] Methods for manufacturing various pharmaceutical formulations containing a certain amount of active ingredient are known and will be apparent to those skilled in the art in light of the present disclosure.

[0079] As described in the attached examples, the inventors have prepared a specific injectable aqueous formulation containing trehalose that is currently being used in ongoing clinical trials in OPMD patients described in Example 6 below (and can be used for the treatment of other diseases associated with abnormal protein aggregation).

[0080] Since more than 99.5% of trehalose is not absorbed into the bloodstream and an oral dose of trehalose of 50 g / day or more in humans often causes the above diarrhea, fullness, and discomfort symptoms, the most effective way to ensure that an appropriate amount of trehalose actually reaches muscle cells is to avoid extensive intestinal metabolism and administer trehalose in an intravenous (IV) solution.

[0081] The disclosed injectable solution of trehalose can be intravenously administered to OPMD patients at the described dosages once daily, every other day, twice a week, once a week, once every 10 days, once every two weeks, or once a month, for about 75 - 120 minutes, for example 90 minutes, per single administration (injection) (which is determined by the attending physician).

[0082] Based on the well-known pharmacokinetic profile of trehalose, initially, once-weekly, 24-week IV therapy may enable trehalose to enter muscle cells and exhibit its therapeutic effect as previously demonstrated in animal models. The amount to be administered by once-weekly injection is based on known safety tests and the clearance and metabolic rates from blood that allow trehalose to reach muscle cells before being removed from the blood.

[0083] The dosage is calculated based on the expected plasma and tissue distribution of trehalose, the effective concentration of trehalose in diseased cells, and the known safe dosages in multiple animal studies.

[0084] Once-weekly administration after initial treatment can be chronic, at specified time points, for several weeks or dozens of weeks, for example an additional 48 consecutive weeks, i.e., a total of 72 weeks of treatment, or the administration can be periodic.

[0085] In one aspect, the molality osmotic concentration of the disclosed aqueous trehalose injectable pharmaceutical formulation is about 280 - about 330 mOsm / Kg.

[0086] As described above, the inventors have shown that trehalose is successfully delivered to plasma and muscle by parenteral administration, specifically IV administration. Thus, in one aspect, the administration of trehalose or a pharmaceutical formulation containing it can be any of intravenous, intramuscular, and intraperitoneal administrations.

[0087] In one aspect, the purified trehalose is substantially free of contaminants such as organic solvents (e.g., ammonia, acetonitrile, acetamide, or alcohol), TFA, ether, or other impurities used in the process that result from its enzymatic production process. In this context, being "substantially" free of contaminants means that the amount of any residual peptide contamination from the enzymatic production process at the end of the purification process is preferably less than 0.5%, less than 0.3%, less than 0.25%, less than 0.1%, less than 0.05%, less than 0.04%, less than 0.03%, less than 0.02%, less than 0.01%, less than 0.005%, less than 0.003%, or less than 0.001% of the total weight of the trehalose. The content of the contaminants can be determined by conventional methods such as chromatography.

[0088] In certain aspects, the residual solvents in the purified trehalose are below the limits set in the ICH guidelines, e.g., IMPURITIES: GUIDELINE FOR RESIDUAL SOLVENTS Q3C(R5) (available at http: / / www.tga.gov.au / pdf / euguide / ich822602006.pdf)

[15] ). For example, the purified trehalose contains <5000 ppm ethanol (e.g., <140 ppm) and / or <3000 ppm methanol.

[0089] Endotoxins (also known as lipopolysaccharides (LPS) and lipoglycans) are large molecules composed of lipids and polysaccharides, consisting of covalently linked O antigen, outer core, and inner core, found in the outer membrane of Gram-negative bacteria and causing a strong immune response in animals. Endotoxins are well-known contaminants in substances purified using bacterial systems, and removing them is extremely important in the safety of using therapeutic agents containing such substances.

[0090] As shown in the examples, the formulations of the present disclosure contain less than 5 endotoxins per kg of the body weight of the patient to whom the formulation is administered per hour of administration. Thus, in one aspect, the pharmaceutical formulation described herein contains less than 0.74 endotoxin units / ml solution.

[0091] The therapeutically effective amount described herein depends on many factors, varies from subject to subject, and can be determined by the judgment well known to those of ordinary skill in the art of the present invention (e.g., a skilled physician). Such factors include the severity of the symptoms, the age, weight, and general condition of the patient, and the judgment of the attending physician.

[0092] Generally, the term "therapeutically effective amount" means an amount of trehalose that is non-toxic and sufficient to produce the desired effect, i.e., the treatment or alleviation of a disease associated with abnormal protein aggregation and / or inclusion body formation in muscle cells, neurons, and other cells, or extracellular compartments.

[0093] The term "therapeutically effective amount or a pharmaceutical composition containing the same" should be understood as the amount of trehalose contained in the administered formulation. The amount of the formulation itself can vary depending on the amount of trehalose contained therein (i.e., the trehalose concentration in the formulation is from about 0.1% (w / v) to about 50% (w / v), and the maximum endotoxin level in the administered formulation is less than 5 endotoxin units / kg body weight / hour).

[0094] The therapeutically effective amount of trehalose represents from about 1 g to about 100 g per single daily injection and an amount of up to 1 g / kg of the treated human subject's body weight per day. The amount can vary considerably. Thus, the range can be from 10, 20, 50, 75, 100, 150, 200, 300 mg / Kg body weight / day to 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, and 1000 mg / Kg body weight / day, respectively. Thus, in another aspect, the therapeutically effective amount of trehalose described herein is from about 1 g to about 100 g per single injection and up to about 1 g / kg of the subject's body weight per day.

[0095] The terms "injection" and "infusion" are used interchangeably and represent parenteral administration of the formulations described herein.

[0096] A therapeutically effective amount of trehalose or a pharmaceutical composition containing the same can be administered parenterally, either as a single dose or as multiple doses (which may be the same or different) at a single dose per day or single administration / day, or the multiple doses can be administered parenterally by continuous administration once a day, once a week, once a month or more.

[0097] The above therapeutically effective amount of trehalose or pharmaceutical composition can be administered IV, IP, or IM as a single dose or as multiple doses (which may be the same or different) at a single dose per day or single administration / day, or if desired as multiple doses at multiple administrations per day (which may be the same or different), or further if desired as equal doses, or gradually increasing doses, or gradually decreasing doses, or further in a chronic or regular dosing schedule.

[0098] The optimal dosing schedule can be calculated from measurements of the amount of drug accumulation in the patient's body. The optimal dose can be determined by the method of administration and the rate of repetition. Generally, the dose is calculated according to body weight and can be administered once, or once a day, once a week, once a month, once a year, or once every few years. One of ordinary skill in the art can easily estimate the rate of repetition of administration based on the measured residence time and concentration of the inventive composition in body fluids or tissues. After successful treatment, it may be desirable for the patient to receive maintenance therapy by administering the inventive composition at a maintenance dose once a day, once or more than once to avoid recurrence of the disease state.

[0099] In one aspect, a therapeutically effective amount of trehalose or a pharmaceutical formulation containing the same described herein is administered chronically.

[0100] As used herein, the term "chronically" refers to a dosing schedule that administers at a predetermined frequency. Thus, administration can be based on a dosing schedule of once daily, once a week, or once a month, for example, by single or multiple parenteral administrations (especially intravenous administration). In one embodiment, the frequency described herein is once a day to once a month, i.e., a therapeutically effective amount of the trehalose or a pharmaceutical formulation containing the same is administered at a frequency of once a day to once a month.

[0101] In other embodiments, the frequency described herein is daily (once a day), i.e., seven times a week, or six, five, four, or three times a week, two or one time a week, once every two weeks, once every three weeks, or once a month.

[0102] In certain embodiments, the frequency described herein is once a week. In other embodiments, a therapeutically effective amount of trehalose or the trehalose contained in the pharmaceutical formulation is administered by single injection once a week. In the foregoing and other embodiments, a therapeutically effective amount of trehalose or the trehalose contained in the pharmaceutical formulation is administered once a day at a trehalose of about 10 mg / kg / day to about 1 g / kg / day.

[0103] As described in the attached examples, in an ongoing clinical trial, a pharmaceutical formulation containing aqueous injectable trehalose is intravenously administered once a week at 8, 15, or 30 g / subject. Thus, in the foregoing and other embodiments, a therapeutically effective amount of trehalose or a pharmaceutical formulation containing the same is intravenously administered as a single dose of about 5 to about 35 g of trehalose, and this can be administered once a day, every other day, twice a week, once a week, once every two weeks, once every three weeks, or once a month. In a further embodiment, a therapeutically effective amount of trehalose or the trehalose contained in the pharmaceutical formulation is administered as a single dose of 5, 8, 15, 30, 40, or 50 g of trehalose.

[0104] In another aspect, a therapeutically effective amount of trehalose or a pharmaceutical formulation containing the same is formulated into an injectable solution such that the rate of administration results in a maximum endotoxin level of less than 5 endotoxin units / kg body weight / hour.

[0105] The term "rate of administration" means the infusion rate (i.e., the rate of administration).

[0106] In a further aspect, a therapeutically effective amount of trehalose or a pharmaceutical formulation containing the same is formulated for intravenous administration such that the administration is completed within less than about 75 to 120 minutes, for example within 90 minutes, though not limited thereto. Also, the administration period depends on the volume / amount of the disclosed injectable trehalose formulation to be administered.

[0107] Accordingly, in one aspect, the administration of the therapeutically effective amount of trehalose contained in the pharmaceutical formulation formulated for intravenous administration is completed within about 75 to about 120 minutes, particularly within less than 90 minutes. In a further aspect, the administration of trehalose or a pharmaceutical formulation containing the same includes a dosing schedule of trehalose or a pharmaceutical formulation containing the same at equal doses, or at gradually increasing doses, or at gradually decreasing doses.

[0108] In one aspect, a therapeutically effective amount of trehalose or a pharmaceutical formulation containing the same is administered periodically.

[0109] The term "periodically" means that the administration of trehalose can be carried out according to a continuous chronic dosing schedule described herein, which can be temporally separated by periods of "no treatment" (i.e., "drug holiday", i.e., when the administration of the active substance to the patient described herein is stopped for a certain period) depending on the patient's condition.

[0110] The determination of the effectiveness of the treatment described herein can be carried out by any method known to those skilled in the art. For example, when the disease to be treated is OPMD, the effectiveness of the treatment can be carried out in relation to any known method for diagnosing or treating OPMD. The alleviation of one or more signs or symptoms of OPMD indicates that the compound provides a clinical benefit.

[0111] Thus, the disease of the subject matter of the present disclosure is OPMD. In that case, the effectiveness of the treatment described herein can be determined by monitoring the patient's weight, having the patient drink 80 ml of ice-cold water, recording the time (seconds) it takes to completely consume this volume (the "water drinking test"), and performing a fiberoptic endoscopic evaluation of swallowing (FEES), a useful adjunct for testing dysphagia. The FEES procedure involves the clinician inserting a flexible fiberoptic endoscope nasally into the patient's hypopharynx so that the structures of the larynx and pharynx can be clearly visualized. Next, the patient is made to perform various tasks (e.g., food and liquid boluses) to evaluate the sensory and motor status of the pharyngeal and laryngeal mechanisms. Information obtained from this test includes the ability to protect the airway, the ability to maintain airway protection for several seconds, the ability to initiate swallowing promptly into the hypopharynx without spillage of the material, the timing and direction of bolus movement in the hypopharynx, the ability to clear the bolus during swallowing, the presence of retention and residue of material in the hypopharynx, the timing of bolus flow and airway protection, the sensitivity of the pharyngeal / laryngeal structures, and the effect of the anatomical structures on swallowing.

[0112] Further tests for evaluating the effectiveness of OPMD treatment include the SWAL-QOL test (a 44-item method in which patients are asked to rate various factors of about 10 quality-of-life concepts related to swallowing on a 5-point scale) developed to objectively measure the patient's swallowing evaluation ability, and muscle strength evaluation (evaluation of proximal muscle weakness including the "stairs climbing" test, the "30-second sit-to-stand switch" test, the "30-second weight arm raise" test, etc.).

[0113] The term "subject in need thereof" as used herein refers to a subject suffering from a disease or disorder described herein, i.e., a disease associated with abnormal protein aggregation and / or inclusion body formation in muscle cells, neurons, and other cells, or the extracellular compartment.

[0114] In one aspect, the present disclosure provides a formulation comprising trehalose or a pharmaceutically acceptable derivative thereof, formulated such that trehalose (or its derivative) or at least a portion of trehalose or its derivative is sustained and / or controlled released and formulated to provide immediate release upon administration of the portion of trehalose or its derivative to a patient.

[0115] In one aspect, the effective serum level of the active ingredient, trehalose or its derivative, is achieved within about 10 to about 20, or 30, or 40, or 50, or 60 minutes after trehalose administration. In one aspect, the effective serum level of the active ingredient is achieved within about 5 to about 20, or 30, or 40, or 50, or 60 minutes after trehalose administration. In one aspect, the effective serum level of the active ingredient is achieved within about 20 to about 20, or 30, or 40, or 50, or 60 minutes after trehalose administration. In one aspect, the effective serum level of the active ingredient is achieved within about 5, 10, 15, 20, 30, 40, 50, or 60 minutes after trehalose administration.

[0116] The inventors have developed innovative approaches for administering trehalose based on parenteral routes. These approaches provide a rational design of a delivery system (e.g., a formulation) with desired properties based on a careful selection of carriers for the intended delivery system, such as a suitable surfactant / cosurfactant composition, or micro / nanoparticles (e.g., liposomes or nanoliposomes), or an active ingredient-encapsulating polymer, or other additives, or excipients.

[0117] Parenteral routes include subcutaneous and transdermal (diffusion through intact skin) administration. In one aspect, the formulations of the present invention are administered by invasive treatment methods such as intravenous administration, intramuscular administration, and the like.

[0118] Another method of administration can use red blood cells loaded with a high amount of trehalose known in the art. It is currently known that loading red blood cells with a high amount of trehalose increases their survival rate. Such red blood cells highly loaded with trehalose can serve as a vehicle for intravenous delivery of trehalose to a subject in need according to the present disclosure.

[0119] A safe and efficient delivery system is required for administering trehalose in pharmaceutical applications. The present disclosure provides a delivery system (e.g., a formulation for parenteral administration) for safely delivering various substances due to its specific physicochemical properties. The delivery system significantly enhances the efficiency and amount of trehalose absorption based on its unique physicochemical properties, enabling delivery of lower concentrations or amounts of an active substance to a subject in a biologically active form. The delivery system brings the active substance into direct proximity to the tissue, resulting in a rapid or nearly rapid effect of trehalose on the subject in need.

[0120] Accordingly, in one aspect, the present invention provides a pharmaceutical delivery system for improving the administration of trehalose or a physiologically active derivative thereof, comprising the trehalose or a physiologically active derivative thereof as an active ingredient in a suitable carrier for rapid recovery of alleviation of symptoms of a disease in a treated subject.

[0121] In one aspect, the drug delivery system can provide the active substance in a controlled release form. In one aspect, the drug delivery system of the present invention may further comprise at least one additional pharmaceutically active agent.

[0122] The delivery system of the present disclosure generally can include a buffer, a substance for adjusting its osmolality, and optionally one or more pharmaceutically acceptable carriers, excipients, and / or additives known in the art. Supplementary pharmaceutically acceptable active ingredients can also be incorporated into the composition. The carrier can be, for example, a solvent or dispersion medium suitable for parenteral administration compositions including water, ethanol, polyols (such as glycerol, propylene glycol, and liquid polyethylene glycol, etc.), suitable mixtures thereof, and vegetable oils. Appropriate fluidity can be maintained, for example, by using coatings such as lecithin, by maintaining the required particle size in the case of dispersions, and by using surfactants.

[0123] As described above, the present trehalose delivery system can be administered in controlled, sustained, or delayed release formulations. Any controlled or sustained release method known to those skilled in the art, such as that described in Langer 1990

[16] , can be used with the formulations and methods of the subject matter of the present disclosure. Such methods include administering a sustained release composition or a coated implantable pharmaceutical device to continuously deliver a pharmaceutically effective dose of the composition of the present invention to the subject of such method. Sustained release can also be achieved using patches designed and formulated for this purpose. Controlled or sustained release compositions include formulations of lipophilic depots (such as fatty acids, waxes, oils). Particle compositions coated with polymers (such as poloxamers or poloxamines) are also included in the present invention. Sustained release formulations, devices, or any topical formulations can further include compositions for stabilizing the composition or for penetrating biological barriers such as the skin or mucosa. Typical additional components can include any physiologically acceptable surfactant or solvent, such as dimethyl sulfoxide (DMSO).

[0124] In certain embodiments, trehalose in the compositions of the present invention can be formulated to provide sustained or controlled release over a period of at least 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 hours. In certain embodiments, trehalose in the compositions of the present invention can be formulated to provide sustained or controlled release over a period of about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 hours. In certain embodiments, trehalose in the compositions of the present invention can be formulated to provide sustained or controlled release over a period of about 0.5 or 1 or 2 or 3 or 4 hours to about 5, 6, 7, 8, 9, 10, 11, or 12 hours. In certain embodiments, trehalose in the compositions of the present invention can be formulated to provide sustained or controlled release over a period of about 5 or 6 or 7 or 8 hours to about 9, 10, 11, or 12 hours. In certain embodiments, trehalose in the compositions of the present invention can be in an immediate, rapid burst release form.

[0125] In certain embodiments, trehalose in the compositions of the present invention can be formulated to release 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 99, 99.5, or 100% or less of the total trehalose in about 0.5, 1, 2, 3, 4, 5, 6, 7, or 8 hours. In certain embodiments, trehalose in the compositions of the present invention can be formulated to release 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 99, 99.5, or 100% or more of the total trehalose in about 0.5, 1, 2, 3, 4, 5, 6, 7, or 8 hours.

[0126] In certain embodiments, the trehalose in the compositions of the present invention can be in a sustained or slow release or a combination of immediate or rapid or burst release forms. In certain embodiments, the relative ratio of sustained or slow release trehalose to immediate or rapid or burst release trehalose can be, for example, 1 to 99, 5 to 95, 10 to 90, 15 to 85, 20 to 80, 25 to 75, 30 to 70, 35 to 65, 40 to 60, 45 to 55, 50 to 50, 55 to 45, 60 to 40, 65 to 35, 70 to 30, 75 to 25, 80 to 20, 85 to 15, 90 to 10, 95 to 5, or 99 to 1.

[0127] In certain embodiments, a polymeric material is used to sustain or control the release of trehalose. In certain embodiments, the type and amount of the polymeric material used have a significant impact on the release rate of trehalose from the compositions and delivery systems of the present invention. Examples of polymers include both hydrophobic and hydrophilic polymers. Examples of hydrophobic polymers include, but are not limited to, ethyl cellulose, and other cellulose derivatives, fats such as glycerol palmito-stearate, beeswax, glyceryl wax, castor wax, carnauba wax, glycerol monostearate, or stearyl alcohol, hydrophobic polyacrylamide derivatives, and hydrophobic methacrylic acid derivatives, and mixtures of these polymers. Examples of hydrophilic polymers include, but are not limited to, hydrophilic cellulose derivatives such as methyl cellulose, hydroxypropyl methyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, carboxymethyl cellulose, sodium carboxymethyl cellulose, and hydroxyethyl methyl cellulose polyvinyl alcohol, polyethylene, polypropylene, polystyrene, polyacrylamide, ethylene vinyl acetate copolymer, polyacrylate, polyurethane, polyvinyl pyrrolidone, polymethyl methacrylate, polyvinyl acetate, polyhydroxyethyl methacrylate, and mixtures of these polymers. Additionally, any mixture of one or more hydrophobic polymers and one or more hydrophilic polymers may also be used if desired.

[0128] The trehalose included in the compositions and delivery systems of the present invention can be encapsulated in liposomes, micro- and nanoparticles.

[0129] In certain embodiments, the polymeric materials used in the compositions and delivery systems of the present invention are microcrystalline cellulose, such as "Avicel PH 101" (manufactured by FMC BioPolymer). Alternatively, the polymeric materials used in the compositions and delivery systems of the present invention are hydroxypropylmethylcellulose, such as "Methocel" (manufactured by Shin-Etsu Chemical Co.). In certain embodiments, the polymeric materials used in the compositions and delivery systems of the present invention are ethylcellulose, such as "Ethocel®" (manufactured by The Dow Chemical Company). In certain embodiments, the polymeric materials used in the compositions and delivery systems of the present invention are acrylic polymers, such as "Eudragit RS®" (manufactured by Rohm GmbH). In certain embodiments, the polymeric materials used in the compositions and delivery systems of the present invention are colloidal silicon dioxide, such as "Aerosil®" (manufactured by Degussa). In certain embodiments, the polymeric materials used in the compositions and delivery systems of the present invention are poly(vinyl acetate), such as "Kollicoat SR" (manufactured by BASF). In certain embodiments, the polymeric materials used in the compositions and delivery systems of the present invention are ethyl acetate and vinyl acetate solutions, such as "Duro-Tak" (manufactured by Delasco Dermatologic Lab & Supply, Inc.).

[0130] In one aspect, the delivery system of the present invention includes a delivery device. In one aspect, the composition of the present invention is delivered by an osmotic pressure method at a controlled rate, such as by an osmotic pump. The system can be constructed by coating an osmotically active substance with a rate-regulating semipermeable membrane. This membrane may include holes of a size important for the delivery of the substance. The dosage form after contact with the liquid absorbs water at a rate determined by the liquid permeability of the membrane and the osmotic pressure of the core formulation. This osmotic absorption of water disperses at a controlled rate from the delivery holes of the membrane, resulting in the formation of a saturated solution of the active substance in the core.

[0131] In one aspect, the composition of the present invention is delivered using biodegradable microparticles. In one aspect, the system for producing the microparticles consists of an organic phase composed of a volatile solution in which the polymer is dissolved and an aqueous phase composed of the substance to be encapsulated emulsified therein. In one aspect, biodegradable polymers that can be used for the microparticle matrix include polylactic acid (PLA) or a copolymer of lactic acid and glycolic acid (PLAGA). The PLAGA polymer is gradually hydrolyzed into its monomer components and is easily removed from the body by natural life processes.

[0132] The preparation may also include an absorption promoter and other desired components. Examples of absorption promoters include, but are not limited to, cyclodextrin, phospholipid, chitosan, DMSO, Tween, Brij, glycocholate, saponin, fusidate, and energy-based absorption promoting devices.

[0133] Desired components present in the dosage form include, but are not limited to, diluents, binders, lubricants, binders, lubricants, surfactants, coloring agents, flavoring agents, buffering agents, preservatives, stabilizers, and the like.

[0134] Diluents (also referred to as "excipients") include, for example, dicalcium phosphate dihydrate, calcium sulfate, lactose, cellulose, kaolin, mannitol, sodium chloride, dried starch, hydrolyzed starch, silicon dioxide, colloidal silica, titanium oxide, alumina, talc, microcrystalline cellulose, and powdered sugar. Diluents for administration in liquid form include, for example, ethanol, sorbitol, glycerol, water, and the like.

[0135] A binder is used to impart adhesiveness to the preparation. Suitable binder substances include, but are not limited to, starches (including corn starch and pregelatinized starch), gelatin, sugars (including sucrose, glucose, dextrose, lactose, and sorbitol), polyethylene glycol, waxes, natural and synthetic gums such as acacia, tragacanth, sodium alginate, cellulose, and Veegum, and synthetic polymers such as polymethacrylate and polyvinylpyrrolidone.

[0136] A lubricant is used to facilitate manufacture. Examples of suitable lubricants include magnesium stearate, calcium stearate, stearic acid, glyceryl behenate, and polyethylene glycol.

[0137] The surfactant can be an anionic, cationic, amphoteric, or nonionic surfactant, but anionic surfactants are preferred. Suitable anionic surfactants include, but are not limited to, those containing carboxylic acids, sulfonic acids, and sulfate ions combined with cations such as sodium, potassium, and ammonium ions. Particularly preferred surfactants include, but are not limited to, long alkyl chain sulfonates, and alkylaryl sulfonates, such as sodium dodecylbenzenesulfonate; dialkyl sodium sulfosuccinates, such as sodium bis-(2-ethylhexyl)-sulfosuccinate; and alkyl sulfates, such as sodium lauryl sulfate. Stabilizers, such as antioxidants, include, but are not limited to, propyl gallate, sodium ascorbate, citric acid, calcium metabisulfite, hydroquinone, and 7-hydroxycoumarin.

[0138] Optionally, the compositions of the present invention may also contain small amounts of non-toxic auxiliary substances such as wetting agents or emulsifiers, preservatives, etc.

[0139] As described, any composition of the present invention can be used alone or in combination with one or more additional therapeutic agents to treat the disease from which the subject to be treated is suffering. The amounts of both the compounds and the additional therapeutic agents that can be combined with the carrier substances for manufacturing the composition or delivery system will vary depending on the recipient to be treated and the particular method of administration. In one embodiment, the compositions of the present invention should be formulated so as to be able to administer trehalose at a dose of 0.01 - 1 g / kg body weight / day. The dose of trehalose depends on the patient's condition and disease and the desired daily dose. In human therapy, the daily dose can be 10 - 3000 mg, for example, 10, 15, 20, 30, 40, 50, 75, 100, 200, 300, 400, 500, 600, 700, 800, 900, or 1000 mg. These amounts can be divided into 2 - 3 doses per day that are less frequent.

[0140] In certain embodiments, the active ingredients in the compositions and delivery systems of the present invention can act synergistically in combination with each other and can further act synergistically in the presence of additional therapeutic agents. Thus, the amounts of the compounds and additional therapeutic agents in such compositions will be less than the amounts required in monotherapy using only the therapeutic agent.

[0141] The toxicity and therapeutic efficacy of the formulations described herein can be determined by standard pharmaceutical methods using cell cultures or experimental animals, e.g., the maximum tolerated dose (MTD), the effective dose which achieves 50% of the maximum response, i.e., the ED 50 , and the therapeutic index (TI), which is the ratio of the ED 50 to the MTD, using methods that are well known in the art. Clearly, formulations with high TI are the most preferred formulations herein, and preferred dosing regimens are those that maintain the plasma level of trehalose at or above the minimum concentration required to maintain the desired therapeutic effect. Of course, the dosage also depends on many factors, including the intended site of delivery, the route of administration, and other relevant factors known to the prescribing physician.

[0142] As noted above, the trehalose or any formulation containing the same described herein can be used alone or in combination with one or more additional therapeutic agents to treat a disease with which a subject to be treated is afflicted. The amounts of both the compounds and additional therapeutic agents that can be combined with the trehalose or any formulation containing the same will vary depending on the subject to be treated, the particular disease, and the method of administration.

[0143] In certain embodiments, the active ingredients in the compositions and delivery systems of the present invention can act synergistically when combined with each other and can further act synergistically in the presence of additional therapeutic agents. Thus, the amounts of the compounds and additional therapeutic agents in such compositions will be less than those required for monotherapy using only the therapeutic agent.

[0144] In another aspect of that situation, the subject matter of the present disclosure is a method for treating or alleviating a disease associated with abnormal protein aggregation and / or inclusion body formation in muscle cells, neurons, and other cells, or extracellular compartments of a human subject in need thereof, or at least one related symptom thereof, the method comprising parenterally administering to the human subject in need thereof a therapeutically effective amount of trehalose or a pharmaceutical formulation comprising the same. Provided is trehalose or a pharmaceutical formulation comprising the same for use in such a method.

[0145] In yet another aspect of that situation, the subject matter of the present disclosure is an aqueous pharmaceutical formulation comprising a therapeutically effective amount of trehalose as the sole active ingredient, having a pH of about 4.5 to 7.0, less than 0.74 endotoxin units / ml, and being suitable for parenteral administration. Provided is such a pharmaceutical formulation.

[0146] In one aspect, the aqueous pharmaceutical formulation described herein may be suitable for intravenous, intramuscular, or intraperitoneal administration. In other aspects, the aqueous pharmaceutical formulation described herein is suitable for intravenous, intramuscular, or intraperitoneal administration. As described above, the aqueous pharmaceutical formulation described herein may further comprise at least one pharmaceutically acceptable additive, carrier, excipient, or diluent, if desired.

[0147] In a further aspect, the aqueous pharmaceutical formulation described herein has a trehalose concentration of about 0.1% (w / v) to about 50% (w / v). In a more specific aspect, the aqueous pharmaceutical formulation described herein has a trehalose concentration of about 10% (w / v).

[0148] In one aspect, the aqueous pharmaceutical formulation described herein has an osmolality of about 280 to 330 mOsm / kg. In other aspects, the aqueous pharmaceutical formulation described herein is administered at a frequency of once a day to once a month.

[0149] The aqueous pharmaceutical formulation described herein has a therapeutically effective amount of trehalose of about 1 g to about 100 g per daily injection and may be up to about 1 g / kg of the subject body weight / day.

[0150] In another aspect, the aqueous pharmaceutical formulation described herein is administered once daily with trehalose at about 10 mg / kg / day to about 1 g / kg / day. In a further aspect, the aqueous pharmaceutical formulation described herein is administered at a frequency of once daily to once monthly with trehalose at a dose of about 5 to about 35 g.

[0151] In a more specific aspect, the aqueous pharmaceutical formulation described herein is administered once daily, every other day, twice a week, once a week, once every two weeks, once every three weeks, or once a month. In a still more specific aspect, the aqueous pharmaceutical formulation described herein has a dose of 5, 8, 15, 30, 40, or 50 g.

[0152] In a further aspect, the aqueous pharmaceutical formulation described herein is such that the administration rate is such that the maximum endotoxin level is less than 5 endotoxin units / kg body weight / hour. The aqueous pharmaceutical formulation of the subject matter of the present disclosure is suitable for intravenous administration, and the administration may be completed within about 75 to about 120 minutes, particularly less than 90 minutes.

[0153] In one aspect, the aqueous pharmaceutical formulation described herein is for treating a disease associated with abnormal protein aggregation and / or inclusion body formation in muscle cells, neurons, and other cells, or the extracellular compartment of a human subject in need thereof, or for alleviating the associated signs or symptoms.

[0154] Furthermore, the subject matter of the present disclosure provides a kit comprising: (a) pharmaceutically acceptable trehalose or an active derivative thereof; (b) at least one pharmaceutically acceptable additive, carrier, excipient, and diluent; (c) means for producing an injectable aqueous solution of the trehalose by mixing the trehalose with at least one of the additive, carrier, excipient, and diluent; (d) means for parenterally administering the injectable solution to a patient in need thereof; (d) instructions for use.

[0155] The injectable aqueous solution of trehalose can be produced by any method well known in the art, for example, any of the methods described in the attached examples of this specification. The parenteral administration of the injectable solution described herein is well known to those skilled in the art.

[0156] As used herein, the term "about" indicates a value that can deviate up to 1%, more specifically 5%, more specifically 10%, more specifically 15%, and in some cases up to 20% higher or lower than the value being referred to, and the deviation range includes integer values and non-integer values that constitute a continuous range where appropriate.

[0157] The present invention will be further described in the following examples, which do not limit the scope of the present invention described in the claims.

[0158] The various aspects and aspects of the present invention described in the above and following claims are experimentally supported by the following examples.

[0159] As disclosed and described, the present invention is not limited to the specific examples, methods, steps, and compositions disclosed herein, and it should be understood that such methods, steps, and compositions can vary somewhat. Also, since the scope of the present invention is limited only by the appended claims and their equivalents, the terms used herein are used only to describe specific aspects and are not intended to be limiting.

[0160] It should be noted that the singular forms "a", "an", and "the" used in this specification and the appended claims include plural referents unless the context clearly indicates otherwise.

[0161] The following examples are representative techniques used by the inventors to implement aspects of the present invention. These techniques are typical of the preferred embodiments for implementing the present invention, but those skilled in the art should recognize that many modifications can be made without departing from the spirit and intended scope of the present disclosure.

[0162] (Example) Without further elaboration, it is believed that one of ordinary skill in the art can, using the preceding description, utilize the present invention to its fullest extent. Accordingly, the following preferred specific embodiments are illustrative only and should not be construed as limiting the present invention in any way.

[0163] Standard molecular biology protocols known in the art that are not specifically described herein generally follow substantially the description in Sambrook & Russell, 2001. Standard pharmaceutical chemistry methods known in the art generally follow essentially the "Comprehensive Medicinal Chemistry" series (published by Pergamon Press) by various authors and editors.

Example

[0164] Trehalose Trehalose dihydrate was obtained from two different commercial sources (Hayashibara and Pfanstiehl). The samples were kept at room temperature until analyzed and analyzed by an acceptable method to verify the details. The results are shown in Table 1 below.

Table 1

[0165] As shown in Table 1 above, the trehalose dihydrate preparations to be tested are suitable for common sense regarding the use of trehalose in pharmaceutical compositions, i.e., they contain 97.0 - 102.0% w / w of the active ingredient (trehalose), the concentration of any peak eluting before trehalose is less than 0.5% w / w, glucose is less than 0.5% w / w, and any peak eluting after trehalose is less than 0.5% w / w. The trehalose used was a commercially available trehalose dihydrate containing 97.0 - 102.0% w / w of the active ingredient (trehalose), less than 0.5% w / w of glucose, and 1% of other impurities.

Example

[0166] Manufacture of Trehalose Solution for IV Injection A preparation containing trehalose is produced by dissolving trehalose dihydrate in water under aseptic conditions, and the resulting clear and colorless liquid is analyzed to identify any impurities or contaminants.

[0167] [Table 2] Table 2 Production of Trehalose Preparation The preparation is sterile, contains 10% trehalose, has a total impurity concentration of 0.1%, a pH of 6.2, an osmolality of 289 mOsm / kg, and less than 0.1 EU / ml of endotoxin.

Example

[0168] Preclinical Pharmacokinetics Study Plasma and muscle concentrations of trehalose in male Sprague-Dawley (SD) rats were measured after intravenous bolus (IV) and oral gavage (PO) administration. All applicable parts of the study were reviewed in accordance with the following regulations and guidelines regarding animal care and welfare: "Guide for the Care and Use of Laboratory Animals", National Research Council ILAR, 1996 revision, reported by AAALAC International (Association for Assessment and Accreditation of Laboratory Animal Care International) and NIH guidelines. This study included 42 SD rats (male, body weight 250 - 350 g, the Shanghai SLAC Laboratory Animal Co. Ltd.). The animals were administered the trehalose preparation (200 mg / mL of trehalose dihydrate in sterile water) intravenously or orally at a dose of 5 ml / kg, achieving a nominal dose of 1 gr / kg. Blood samples were collected after administration of each dose to obtain plasma. Muscle samples (hind limb muscle) were collected and homogenized. Trehalose concentrations in plasma and muscle homogenate samples were analyzed by an appropriate bioanalytical LC / MS / MS method.

[0169] Pharmacokinetic Data Analysis The plasma concentration data of trehalose were subjected to non-compartmental pharmacokinetic analysis using the WinNonlin software program (version 6.3, Pharsight, Mountain View, CA). The zero-time intercept concentration (C0), volume of distribution at steady state (Vdss), clearance (Cl), peak plasma concentration (Cmax), and corresponding peak time (Tmax), terminal half-life (T1 / 2), mean residence time from zero to the last time point (MRT)(MRT0-last), MRT from zero to infinity (MRT0-inf), area under the plasma concentration-time curve from zero to the last time point (AUC)(AUC0-last), and AUC from zero to infinity (AUC0-inf) were calculated using the linear / logarithmic trapezoidal formula. Since there was no deviation greater than 5% between the actual sampling time and the nominal sampling time, all pharmacokinetic parameters were calculated using the nominal sampling time. Both the muscle concentration value relative to the plasma concentration and the value of the muscle-to-plasma concentration and AUC ratio (M / P ratio) were calculated.

[0170] Trehalose Concentrations in Plasma and Muscle The pharmacokinetic parameters of trehalose in plasma and muscle after single intravenous or oral administration of a 1000 mg / kg solution of trehalose dihydrate (200 mg trehalose dihydrate / 1 mL sterile water) to male SD rats are shown in Table 3 below.

[0171] The individual and mean plasma concentrations of trehalose after intravenous or oral administration of a 1000 mg / kg solution of trehalose dihydrate (200 mg trehalose dihydrate / 1 mL sterile water) to male SD rats are shown in Table 3 and illustrated in Figure 1.

[0172] The individual and average plasma concentrations of trehalose after single intravenous or oral administration of a 1000 mg / kg solution of trehalose dihydrate (200 mg trehalose dihydrate / 1 mL sterile water) to male SD rats are also shown in Table 3. The comparison of the plasma and muscle concentrations of trehalose after single intravenous or oral administration of a 1000 mg / kg solution of trehalose dihydrate to male SD rats is illustrated in Figures 2-3.

[0173] After single intravenous administration of a 1000 mg / kg solution of trehalose (200 mg trehalose dihydrate / 1 mL sterile water) to male fasted SD rats in Test Groups 1-5, trehalose showed a total clearance (CI) of 17.2 mL / min / kg (about 31.3% of rat liver blood flow (= 55 mL / min / kg)), and the mean elimination half-life (T 1 / 2 ) was 2.07 hours. C0 was 1,370,000 ng / mL. The volume of distribution (V dss ) was 0.685 L / kg. The mean plasma exposure AUC 0-last (48 hr) was 778,000 ng·hr / mL.

[0174] When a solution of trehalose dihydrate (200 mg trehalose dihydrate / 1 mL sterile water) was orally administered to male SD rats in Test Groups 6-10, the maximum plasma concentration of trehalose (C max ) was reached at 0.5 hour (T max = 4,280 ng / mL) after administration. AUC 0-last (3 hr) was 4,520 ng / mL·hr. The absolute bioavailability of trehalose was estimated to be as low as 0.601%.

[0175] The pharmacokinetic properties of trehalose showed that it was rapidly absorbed and reached the peak plasma concentration over time, but the absolute oral bioavailability was very low, and the compound may undergo significant presystemic metabolism.

[0176] After single intravenous administration of a 1000 mg / kg solution of trehalose dihydrate (200 mg trehalose dihydrate / 1 mL sterile water) to male SD rats in Test Groups 1-5, the C of trehalose in musclemax was 3730 ng / mL and was observed at 8 hours after dosing (T max ). The muscle exposure AUC 0-last (48 hr) was 107,000 ng·hr / mL and the elimination half-life was 33.8 hours. The PK parameters of muscle samples from the drug-treated oral groups (Groups 6 - 10) could not be calculated as they were lower than the LLOQ.

[0177] The mean ratio of muscle trehalose concentration to plasma concentration was 2.88 - 3.76 in male SD rats after intravenous administration. The muscle / plasma concentration ratio of orally administered trehalose was lower than the LLOQ and could not be calculated.

[0178] Conclusion Trehalose in plasma and muscle tissues was measured in male SD rats after intravenous or oral administration of a 1000 mg / kg solution of trehalose dihydrate in sterile water. Plasma glucose was also monitored in each sample obtained from the test animals in the drug-treated groups. The following conclusions can be drawn: First, the total clearance (Cl) of trehalose after IV administration was 17.2 mL / min / kg, accounting for approximately 31.3% of hepatic blood flow (a moderate value of hepatic extraction ratio). V dss and T 1 / 2 were 0.685 L / kg and 2.07 hours, respectively. The mean plasma exposure AUC 0-last was 778,000 ng·hr / mL.

[0179] After oral administration, trehalose was rapidly absorbed and T max was observed at 0.50 hours after dosing, but the absolute oral bioavailability was low at 0.601%, suggesting that presystemic metabolism may play an important role. The T 1 / 2 of trehalose was significantly shorter in orally dosed rats compared to the intravenous administration group.

[0180] After IV administration, the mean muscle trehalose concentration / plasma concentration ratio was 2.88 - 3.76 in male SD rats. Finally, it was found that rats were sufficiently tolerant to trehalose dihydrate at a given dose.

[0181]

Table 3

[0182] Therefore, as shown in Table 3, the trehalose of the IV-administered formulation had a T of 2.07 in plasma 1 / 2 and was more than twice as high as the plasma T obtained with trehalose of the orally administered formulation (0.740 hours). Furthermore, the muscle T obtained with trehalose of the IV-administered formulation 1 / 2 was 33.8 hours. Also, the AUC values obtained in plasma and muscle when the formulation was administered IV were significantly higher than the respective AUC values of the orally administered formulation. 1 / 2

[0183] Furthermore, as shown in Figure 1, the average plasma concentration of trehalose of the IV-administered formulation was higher than the average plasma concentration of trehalose of the orally administered formulation at each time point tested.

[0184] Interestingly, Figure 2 showing the plasma-to-muscle concentration of trehalose of the intravenously administered trehalose formulation indicates that the muscle concentration of trehalose was higher than the plasma concentration. The plasma and muscle concentrations of trehalose were undetectable in the orally administered trehalose formulation.

Example

[0185] Measurement of Endotoxin Level The maximum allowable level of endotoxin in the intravenously administered formulation is recognized to be 5 endotoxin units (EU) / kg body weight / hour (5 EU / kg / hr). The following calculation was performed to determine the theoretical maximum endotoxin level IV / kg body weight / hour (K) of the trehalose formulation (sterile aqueous solution of trehalose dihydrate):

[0186]

Table 4

[0187] As shown in Table 4 above, the endotoxin level / ml in trehalose formulations manufactured using standard solvents (such as water, physiological saline, etc.) is 0.74 EU / ml. Assuming a moderate infusion rate over 75 minutes for body weights of 60, 50, and 40 kg, the endotoxin levels in a 10% (w / v) trehalose formulation are 1.5, 1.8, and 2.2 EU / kg / hr respectively for formulations containing 15 g of trehalose, and 3.0, 3.6, and 4.4 EU / kg / hr respectively for formulations containing 30 g of trehalose.

[0188] Therefore, for body weights of 60, 50, and 40 kg, the endotoxin levels will be 2, 2.4, and 3 EU / kg / hr respectively for formulations containing 15 g of trehalose (in 150 ml of solvent), and 4, 4.8, and 6 EU / kg / hr respectively for formulations containing 30 g of trehalose (in 300 ml of solvent).

Example

[0189] Safety Test The safety and tolerance of trehalose were extensively investigated as follows. The median lethal dose (LD 50 ) of trehalose was tested in mice, rats, and dogs. None of the species showed any signs of toxicity or died after oral and intravenous administration. The results are summarized in Table 5 below.

Table 5

[0190] Furthermore, trehalose is recognized as a GRAS substance used in the pharmaceutical industry as a safe food ingredient and a carrier for oral, intraocular, and IV formulations. In various tests, healthy volunteers were orally administered trehalose in the range of 10 - 60 g. No other safety issues were reported except for mild abdominal symptoms (e.g., fullness, flatulence, borborygmus, and occasional diarrhea)

[18] . Trehalose has been used as a protein stabilizer for various commercial protein drugs over a 10-year period, and its safety has been repeatedly established in patient populations with advanced stages of malignant diseases, hemophilia, and related coagulation disorders. These drugs are approved for use over several years and may be administered to patients at a frequency of every 8 hours at 2 - 3 week intervals.

Example

[0191] Treatment of OPMD Patients with Trehalose The potential usefulness of trehalose in alleviating the symptoms of OPMD or delaying the deterioration of OPMD patients is currently being experimentally tested in clinical trials of human patients. This trial is a randomized, double-blind, dose-escalation, and parallel-group dose-controlled trial of the treatment of patients with oculopharyngeal muscular dystrophy (OPMD) type IV with an aqueous injectable solution of trehalose conducted at 3 medical centers.

[0192] Preliminary Phase Screening Period (Week - 4 / Day - 28 to Week 0 / Day 0) The screening evaluation is conducted in two hospital visits within 28 days before the start of treatment as described in the "Evaluation Schedule".

[0193] Treatment Period 1 (Week 1 to Week 24) All eligible patients receive the test treatment once a week. Initially, all eligible patients are administered intravenously a single dose (80 ml solution) of trehalose (8 g) as a 10% (w / v) injectable aqueous solution once a week. Subsequently, the following week, a single dose of trehalose (15 g) as a 10% (w / v) injectable aqueous solution is administered intravenously. After safety review, at the next visit in the following weeks, patients are randomized in a 1:1 ratio (double-blind) to receive 15 g or 30 g of trehalose (aqueous 10% (w / v) solution) for 24 weeks. The first 4 infusions are performed at the clinic under the instruction of an expert. Patients return to the clinic once a month for drug infusion and test evaluation as described in the "Treatment Schedule". All other weekly infusions can be performed at the patient's home or at the clinic.

[0194] Core Phase Treatment Period 2 (Week 25 to Week 72) Continue the once-weekly IV infusion of the aqueous trehalose injectable solution to the patients.

[0195] Follow - up Observation Period (4 weeks after administration) Patients come to the hospital for follow-up observation after treatment 4 weeks after the final dose in the 24th week.

[0196] Test Population Register up to a maximum of 15 adult OPMD patients in the trial at each of the 3 trial centers. A total of at least 42 patients are to be registered. A control group that does not receive treatment can be added.

[0197] Selection Criteria 1. Males and females 2. Aged 18 - 80 years (inclusive) 3. Genetically diagnosed with OPMD 4. Moderate dysphagia (abnormal water drinking test at screening and on the first administration day (before drug administration)) 5. Patients must be ambulatory and able to undergo muscle function and strength assessment. 6. Patients who give written informed consent to participate in this trial. 7. Body mass index (BMI) < 30 kg / m2 8. Female patients who are potentially fertile must have a negative serum pregnancy test at screening. 9. Men and women must consent to acceptable contraception.

[0198] Exclusion Criteria 1. Type 1 or type 2 diabetes 2. Other major diseases, such as renal insufficiency (creatinine clearance <60 ml / min), hepatic insufficiency, and chronic liver diseases (e.g., hepatitis B or C), HIV carriers, tuberculosis, SLE, rheumatoid arthritis, connective tissue diseases. 3. Poorly controlled heart diseases, such as CHF 4. Other neuromuscular diseases 5. Other esophageal dysphagia-related diseases: e.g., gastroesophageal reflux (GERD), esophageal stricture due to mechanical or chemical trauma, infectious diseases (e.g., esophageal candidiasis), drug-induced dysphagia (e.g., bisphosphonates), esophageal rings and membranes, spastic motility disorders of the esophagus 6. History of malignancy 7. History of cervical radiation exposure 8. Pregnant women or women who are currently breastfeeding 9. Obesity (BMI > 30) and related conditions 10. Previous laryngectomy 11. Weight loss of 10% or more in the past 12 months 12. Known allergies to any of the components by injection.

[0199] Investigational Product Route and Dosage Form Administer a 10% (w / v) trehalose IV solution once a week by single injection for 72 weeks. Deliver the solution over approximately 70 to approximately 120 minutes. The initial dose / injection in week 1 is 8 g of trehalose. The dose in week 2 is 15 g and the doses in weeks 3 to 72 are 15 or 30 g.

[0200] Safety and Tolerability The primary safety endpoint is the frequency, severity, and duration of adverse events (AEs) including clinically significant laboratory finding abnormalities after administration of the trehalose injectable solution. Safety is evaluated based on AEs and concomitant medications; physical examinations; and vital signs (before dosing, every 30 minutes during dosing, and for 30 minutes after dosing). Examinations include complete blood count (CBC), electrolytes (Na, K, Cl), BUN, creatinine, glucose, liver function tests (ALT, AST, total bilirubin, direct bilirubin, alkaline phosphatase, and serum albumin), and urinalysis.

[0201] Results of Efficacy / Disease Marker Evaluation The following disease markers are evaluated at specific times: · Deglutition function evaluation score (using video fluoroscopic observation) · SWAL-QOL · Muscle timing function and strength evaluation

[0202] The change compared to baseline is measured for each patient, and the total score change of the treatment group at each predefined efficacy endpoint is statistically analyzed. Body weight and alcohol consumption may be further evaluated. Percutaneous Core Needle Biopsy (PCNB) is performed to obtain muscle fibers for histological examination.

[0203] Pharmacokinetics The pharmacokinetics of trehalose are evaluated in patients at a dose of 15 g or 30 g of trehalose. The blood concentration of trehalose is measured before dosing (within 60 minutes before drug administration) and every 30 minutes for 5 hours after the start of dosing or until the glucose level returns to normal (whichever occurs first). The present invention has been described in detail along with its description, but the above description is illustrative and does not limit the scope of the present invention, which is defined by the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Claims

1. A pharmaceutical formulation comprising a therapeutically effective amount of trehalose, 1. A pharmaceutical formulation for use in a method for treating a disease associated with abnormal protein aggregation and / or inclusion body formation in muscle cells, neurons or in extracellular compartments or for alleviating at least one associated symptom thereof in a human subject in need thereof, the method comprising intravenously administering to the subject a pharmaceutical formulation comprising 0.1% (w / v) to 50% (w / v) trehalose, wherein the formulation has a pH of 4.5-7.0 and contains less than 0.74 endotoxin units / ml; wherein the administration is completed within about 120 minutes; and wherein the disease is amyotrophic lateral sclerosis (ALS),

2. 2. The pharmaceutical formulation for use according to claim 1, wherein the pharmaceutical formulation is an injectable solution for intravenous administration over a period of 90 minutes.

3. 3. The pharmaceutical formulation for use according to claim 1 or 2, wherein the pharmaceutical formulation further comprises at least one pharma- ceutically acceptable excipient, carrier, excipient, or diluent.

4. The pharmaceutical formulation for use according to any one of claims 1 to 3, wherein the concentration of trehalose in the formulation is 10% (w / v).

5. 5. The pharmaceutical formulation for use according to any one of claims 1 to 4, wherein the pharmaceutical formulation comprising trehalose has an osmolality of 280 to 330 mOsm / kg.

6. 6. The pharmaceutical formulation for use according to any one of claims 1 to 5, wherein said therapeutically effective amount of trehalose is between 1g and 100g per single injection and not more than 1g / Kg subject body weight / day.

7. The pharmaceutical preparation for use according to any one of claims 1 to 6, wherein said therapeutically effective amount of the pharmaceutical preparation is administered chronically.

8. 8. The pharmaceutical formulation for use according to any one of claims 1 to 7, wherein the therapeutically effective amount of the pharmaceutical formulation is administered at a frequency between once a day and once a month, optionally wherein the pharmaceutical formulation is administered once a day at 10 mg / kg / day to 1 g / kg / day of trehalose, or wherein the pharmaceutical formulation is administered intravenously in a single dose of 5 to 35 g of trehalose once a day, every other day, twice a week, once a week, once every two weeks, once every three weeks or once a month, or wherein the pharmaceutical formulation is administered in a single dose of 5, 8, 15, 30, 40 or 50 g trehalose.

9. 8. The pharmaceutical formulation for use according to any one of claims 1 to 7, wherein said therapeutically effective amount of the pharmaceutical formulation is administered in a single injection once per week, optionally said therapeutically effective amount of the pharmaceutical formulation is administered intravenously in a single dose of 5 to 35 g trehalose once per day, every other day, twice per week, once per week, once per two weeks, once per three weeks or once per month, or said therapeutically effective amount of the pharmaceutical formulation is administered in a single dose of 5, 8, 15, 30, 40 or 50 g trehalose.

10. 10. The pharmaceutical formulation for use according to any one of claims 1 to 9, wherein the therapeutically effective amount of the pharmaceutical formulation is (i) an injectable solution and the administration rate is such that the maximum endotoxin level is less than 5 endotoxin units / kg subject weight / hour.

11. The pharmaceutical formulation for use according to any one of claims 1 to 10, wherein said administration comprises a regimen of equal doses, or gradually increasing or decreasing doses of the pharmaceutical formulation.

12. The pharmaceutical formulation for use according to any one of claims 1 to 6 or claims 8 to 11, wherein said therapeutically effective amount of the pharmaceutical formulation is administered periodically.

13. The pharmaceutical formulation for use according to any one of claims 1 to 12, wherein the formulation further comprises a trehalase inhibitor.

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