Compositions comprising SGLT inhibitors and uses thereof

Abstract

The present invention relates to compositions comprising an SGLT inhibitor and uses thereof. The present invention provides a composition comprising a pharmaceutical agent capable of inhibiting the activity of SGLT. The present invention further relates to methods of treating and / or ameliorating symptoms associated with cystic fibrosis (CF) comprising administering to a subject (e.g., a human patient) a composition comprising one or more pharmaceutical agents that act as an inhibitor of SGLT activity.

Description

[0001] This application is a divisional application of a Chinese patent application with an application date of September 4, 2020, an application number of 202080076294X, and an invention title of "SGLT Inhibitors and Their Uses". Field of the Invention

[0002] The present invention belongs to the field of pharmaceutical pharmacology. Specifically, the present invention relates to pharmaceutical agents that act as inhibitors of the activity of sodium-glucose cotransporters (SGLTs). The present invention further relates to methods for treating and / or ameliorating symptoms associated with cystic fibrosis (CF), the method comprising administering to a subject (e.g., a human patient) a composition comprising one or more pharmaceutical agents that act as SGLT activity inhibitors. Background Art

[0003] An estimated 70,000 children and adults worldwide suffer from cystic fibrosis (CF). CF is a life-threatening genetic disease caused by mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR) protein. CFTR is a chloride channel expressed in many epithelial cell types. Mutations in the CFTR gene result in abnormal transport of water and electrolytes across the apical cell membrane of many exocrine tissues such as the lung. CFTR gene mutations have been classified into five classes of protein molecular defects: class I, premature termination codons resulting in complete absence of CFTR protein synthesis; class II, maturation arrest and intracellular localization defects (processing block); class III, defects in the activation and regulation of chloride transport function (gating defects); class IV, reduced chloride channel conductance; and class V, reduced CFTR protein synthesis. The most common CFTR mutation is the deletion of the phenylalanine residue at position 508 of the polypeptide chain (mutation F508del, mutant protein F508del-CFTR), which belongs to class II defects. This mutation is present on at least one allele in approximately 90% of CF patients, and almost 50% of the genotyped patients are homozygous for F508del (see, Egan et al., Science, 2004, 304: 600-602). The F508del mutation causes CFTR to fail to be correctly transported to the plasma membrane because protein misfolding retains the protein in the endoplasmic reticulum. Additionally, when the F508del-CFTR protein is correctly localized to the plasma membrane, it also has altered intrinsic chloride channel transport function relative to the wild-type (WT) CFTR protein (see, Dalemans et al, Nature, 1991, 354: 526-528).

[0004] In October 2019, the Food and Drug Administration (FDA) of the United States approved Trikafta, a combination of the CFTR potentiator VX-770 and the CFTR corrector VX-445 and VX-661, which can provide benefits to more than 90% of CF patients. Although the entire field celebrated this milestone achievement 30 years after the discovery of the CFTR gene, the consensus remains that this marks a new beginning rather than the end of the effort to gain a deeper understanding of the disease and develop new and more effective therapeutic agents for all patients, as CF has not been cured.

[0005] Accordingly, there is a need for improved methods and techniques for treating and / or ameliorating CF.

[0006] The present invention addresses this need. SUMMARY OF THE INVENTION

[0007] During the course of developing embodiments of the present invention, experiments were conducted using CF rabbits to examine the beneficial effects of SGLT inhibitor drugs on CF. It has been shown in a CF animal model that SGLT inhibitor drugs such as LX4211 have beneficial effects on CF complications in multiple organ systems, indicating that SGLT inhibitor drugs may provide therapeutic benefits to CF patients with symptoms such as hypokalemia, hyperglycemia, dyslipidemia, hypoalbuminemia, and hypoproteinemia. Additional results show that LX4211 improves glucose tolerance in CF rabbits, LX4211 improves blood chemistry parameters in CF rabbits, LX4211 is beneficial for electrolyte imbalance and lipid metabolism in CF rabbits, LX4211 does not affect weight gain and prolongs the lifespan of CF rabbits. In fact, these results show that SGLT1 is upregulated in human CF airway lineage cells and in many CF-related tissues of CF rabbits; and the SGLT inhibitor LX4211 brings many beneficial effects to CF rabbits.

[0008] Accordingly, the present invention relates to pharmaceutical agents that act as inhibitors of SGLT activity, and methods of treating and / or ameliorating symptoms associated with cystic fibrosis (CF) with such SGLT activity inhibitors.

[0009] In certain embodiments, the present invention provides a composition comprising a pharmaceutical agent capable of inhibiting SGLT activity.

[0010] In certain embodiments, the present invention provides a method for inhibiting SGLT activity in a subject, the method comprising administering to the subject a composition comprising a pharmaceutical agent capable of inhibiting SGLT activity. In some embodiments, the subject is a human subject. In some embodiments, the subject is a human subject suffering from or at risk of suffering from any form and / or mutation associated with CF.

[0011] In certain embodiments, the present invention provides methods for treating, ameliorating, and / or preventing CF in a subject, the methods comprising administering to the subject a composition comprising a pharmaceutical agent capable of inhibiting SGLT activity. In some embodiments, the subject is a human subject. In some embodiments, the subject is a human subject who has or is at risk of having any form and / or mutation associated with CF.

[0012] In certain embodiments, the present invention provides methods for treating, ameliorating, and / or preventing one or more symptoms associated with CF in a subject, the methods comprising administering to the subject a composition comprising a pharmaceutical agent capable of inhibiting SGLT activity. In some embodiments, the subject is a human subject. In some embodiments, the subject is a human subject who has or is at risk of having any form and / or mutation associated with CF. In some embodiments, one or more symptoms associated with CF include, but are not limited to, hypokalemia, hyperglycemia, dyslipidemia, hypoalbuminemia, and hypoproteinemia.

[0013] In certain embodiments, the present invention provides methods for treating, ameliorating, and / or preventing hypokalemia associated with CF in a subject, the methods comprising administering to the subject a composition comprising a pharmaceutical agent capable of inhibiting SGLT activity. In some embodiments, the subject is a human subject. In some embodiments, the subject is a human subject who has or is at risk of having any form and / or mutation associated with CF. In some embodiments, the subject is a human subject who has or is at risk of having hypokalemia associated with CF.

[0014] In certain embodiments, the present invention provides methods for treating, ameliorating, and / or preventing hyperglycemia associated with CF in a subject, the methods comprising administering to the subject a composition comprising a pharmaceutical agent capable of inhibiting SGLT activity. In some embodiments, the subject is a human subject. In some embodiments, the subject is a human subject who has or is at risk of having any form and / or mutation associated with CF. In some embodiments, the subject is a human subject who has or is at risk of having hyperglycemia associated with CF.

[0015] In certain embodiments, the present invention provides methods for treating, ameliorating, and / or preventing CF-related dyslipidemia in a subject, the methods comprising administering to the subject a composition comprising a pharmaceutical agent capable of inhibiting SGLT activity. In some embodiments, the subject is a human subject. In some embodiments, the subject is a human subject who has or is at risk of having any form and / or mutation associated with CF. In some embodiments, the subject is a human subject who has or is at risk of having CF-related dyslipidemia.

[0016] In certain embodiments, the present invention provides methods for treating, ameliorating, and / or preventing CF-related hypoalbuminemia in a subject, the methods comprising administering to the subject a composition comprising a pharmaceutical agent capable of inhibiting SGLT activity. In some embodiments, the subject is a human subject. In some embodiments, the subject is a human subject who has or is at risk of having any form and / or mutation associated with CF. In some embodiments, the subject is a human subject who has or is at risk of having CF-related hypoalbuminemia.

[0017] In certain embodiments, the present invention provides methods for treating, ameliorating, and / or preventing CF-related hypoproteinemia in a subject, the methods comprising administering to the subject a composition comprising a pharmaceutical agent capable of inhibiting SGLT activity. In some embodiments, the subject is a human subject. In some embodiments, the subject is a human subject who has or is at risk of having any form and / or mutation associated with CF. In some embodiments, the subject is a human subject who has or is at risk of having CF-related hypoproteinemia.

[0018] Such methods are not limited to treating specific forms or mutations associated with CF. In some embodiments, the mutation is any mutation associated with class 1A CTFR mutations (e.g., Dele2,3(21kb) and 1717-1G→A). In some embodiments, the mutation is any mutation associated with class 1B CTFR mutations (e.g., Gly542X and Trp1282X). In some embodiments, the mutation is any mutation associated with class 2 CTFR mutations (e.g., Phe508del, Asn1303Lys, and Ala561Glu). In some embodiments, the mutation is any mutation associated with class 3 CTFR mutations (e.g., Gly551Asp, Ser549Arg, and Gly1349Asp). In some embodiments, the mutation is any mutation associated with class 4 CTFR mutations (e.g., Arg117His, Arg334Trp, and Ala455Glu). In some embodiments, the mutation is any mutation associated with class 5 CTFR mutations (e.g., 3272-26A→G, 3849+10kg C→T). In some embodiments, the mutation is any mutation associated with class 6 CTFR mutations (e.g., c.120del123 and rPhe580del).

[0019] In certain embodiments, the present invention provides a kit, the kit comprising (1) a composition comprising a pharmaceutical agent capable of inhibiting SGLT activity, (2) a container, package, or dispenser, and (3) instructions for administration.

[0020] Such compositions, methods, and kits are not limited to a particular type or class of pharmaceutical agent capable of inhibiting SGLT activity. In some embodiments, the pharmaceutical agent capable of inhibiting SGLT activity is a small molecule, an antibody, a nucleic acid molecule (e.g., siRNA, antisense oligonucleotide), or a peptidomimetic.

[0021] In some embodiments, the pharmaceutical agent capable of inhibiting SGLT activity is selected from, for example, phlorizin, canagliflozin ((2S,3R,4R,5S,6R)-2-{3-[5-(4-fluorophenyl)-thiophen-2-ylmethyl]-4-methylphenyl}-6-hydroxymethyl-tetrahydropyran-3,4,5-triol), dapagliflozin ((2S,3R,4R,5S,6R)-2-[4-chloro-3-(4-ethoxybenzyl)phenyl]-6-(hydroxymethyl)-tetrahydro-2H-pyran-3,4,5-triol), empagliflozin ((2S,3R,4R,5S,6R)-2-[4-chloro-3-[[4-[(3S)-oxolan-3-yl]oxyphenyl]methyl]phenyl]-6-(hydroxymethyl)oxane-3,4,5-triol), remogliflozin (5-methyl-4-[4-(1-methylethoxy)benzyl]-1-(1-methylethyl)-1H-pyrazol-3-yl 6-O-(ethoxycarbonyl)-β-D-glucopyranoside), sergliflozin (2-(4-methoxybenzyl)phenyl 6-O-(ethoxycarbonyl)-β-D-glucopyranoside), and tofogliflozin ((1S,3'R,4'S,5'S,6'R)-6-(4-ethylbenzyl)-6'-(hydroxymethyl)-3',4',5',6'-tetrahydro-3H-spiro[2-benzofuran-1,2'-pyran]-3',4',5'-triol hydrate (1:1)), and sotagliflozin (LX4211), or a pharmaceutically acceptable salt thereof. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 Shows that LX4211 restores glucose tolerance in CF rabbits.

[0023] Figure 2 Demonstrates that LX4211 treatment improves glucose tolerance in CF rabbits.

[0024] Figure 3 Demonstrates that LX4211 treatment results in improved survival length in CF rabbits.

[0025] Figure 4 Demonstrates the beneficial effects of LX4211 on electrolyte imbalance and glycolipid metabolism disorders in the CF rabbit model.

[0026] Figure 5 Shows that LX4211 treatment significantly alleviates hypokalemia and hyperglycemia in CF rabbits.

[0027] Figure 6 LX4211 is shown to restore total Chol (cholesterol), CPK (creatine kinase), ALB (albumin), and TPRO (total protein).

[0028] Figure 7 an shows SGLT1 expression in CF rabbit tissues. (ad) mRNA levels in different tissues of CF and WT rabbits. (e) SGLT1 and CFTR protein levels in intestinal (int) and pancreatic (pan) tissues of CF and WT rabbits. (f) SGLT and CFTR protein levels in lungs of WT and CF rabbits. (gn) Immunostaining of SGLT1 (brown) in intestine and pancreas of WT and CF rabbits.

[0029] Figure 8 .SGLT1 protein levels in CF patient-derived airway lineage cells. Left: SGLT1 and CFTR levels in CFBE cells. Right: SGLT1 protein levels in lung organoids of different genotypes (WT / WT, dF / dF, and dF / G551D) with or without forskolin (FSK) stimulation.

[0030] Figure 9 .Urine glucose levels in WT rabbits after Sota treatment.

[0031] Figure 10 AC. Sota treatment regimen and its GTT test effect. (A) Sota treatment regimen. (B) GTT curves of CF rabbits before (red line) and after (blue line) Sota treatment. (C) Summary of the area under the curve (AUC).

[0032] Figure 11 Selected blood chemistry results of CF rabbits treated with Sota (green dots) or not treated with Sota (red dots). Gray box: normal range.

[0033] Figure 12 .Weight (left) and survival curve (right) of CF rabbits treated with Sota.

[0034] Figure 13 . Schematic diagram showing that SGLT1 is upregulated in human CF airway lineage cells and in many CF-related tissues in CF rabbits; and the SGLT inhibitor Sota brings many beneficial effects to CF rabbits. DETAILED DESCRIPTION

[0035] Cystic fibrosis (CF) is a fatal autosomal recessive disease caused by mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR). CF patients mainly suffer from CF lung disease, as well as CF-related liver disease, CF-related diabetes, CF-related gastrointestinal diseases, etc. Electrolyte abnormalities and acid-base disorders are also associated with CF, including hypokalemia and metabolic alkalosis. Sodium-glucose cotransporter (SGLT) inhibitors, including selective SGLT2 inhibitors and dual SGLT1 / 2 inhibitors, have become the mainstream treatment for diabetes. The role of SGLT inhibitors in CF has not been systematically tested.

[0036] Experiments implemented during the process of developing embodiments of the present invention utilize CF rabbits to examine the beneficial effects of SGLT inhibitor drugs on CF. In CF animal models, SGLT inhibitor drugs such as LX4211 are shown to have beneficial effects on CF complications of multiple organ systems, thereby indicating that SGLT inhibitor drugs may provide therapeutic benefits for CF patients with symptoms such as hypokalemia, hyperglycemia, dyslipidemia, hypoalbuminemia, and hypoproteinemia. Additional results show that LX4211 improves glucose tolerance in CF rabbits, LX4211 improves blood chemistry parameters in CF rabbits, LX4211 is conducive to electrolyte imbalance and lipid metabolism in CF rabbits, and LX4211 does not affect weight gain and prolongs the life span of CF rabbits. In fact, these results show that SGLT1 is upregulated in human CF airway lineage cells and in many CF-related tissues in CF rabbits; and the SGLT inhibitor LX4211 brings many beneficial effects to CF rabbits.

[0037] Accordingly, the present invention relates to pharmaceutical agents that function as inhibitors of SGLT activity, and methods of using such inhibitors of SGLT activity to treat and / or ameliorate symptoms associated with cystic fibrosis (CF).

[0038] In certain embodiments, the present invention provides a method for inhibiting SGLT activity in a subject, the method comprising administering to the subject a composition comprising a pharmaceutical agent capable of inhibiting SGLT activity. In some embodiments, the subject is a human subject. In some embodiments, the subject is a human subject suffering from or at risk of suffering from any form and / or mutation associated with CF.

[0039] In certain embodiments, the present invention provides a method for treating, improving and / or preventing CF in a subject, the method comprising administering to the subject a composition comprising a pharmaceutical agent capable of inhibiting SGLT activity. In some embodiments, the subject is a human subject. In some embodiments, the subject is a human subject suffering from or at risk of suffering from any form and / or mutation associated with CF.

[0040] In certain embodiments, the present invention provides a method for treating, improving and / or preventing one or more symptoms associated with CF in a subject, the method comprising administering to the subject a composition comprising a pharmaceutical agent capable of inhibiting SGLT activity. In some embodiments, the subject is a human subject. In some embodiments, the subject is a human subject suffering from or at risk of suffering from the following items: any form and / or mutation associated with CF. In some embodiments, one or more symptoms associated with CF include, but are not limited to, hypokalemia, hyperglycemia, dyslipidemia, hypoalbuminemia, and hypoproteinemia.

[0041] In certain embodiments, the present invention provides a method for treating, improving and / or preventing hypokalemia associated with CF in a subject, the method comprising administering to the subject a composition comprising a pharmaceutical agent capable of inhibiting SGLT activity. In some embodiments, the subject is a human subject. In some embodiments, the subject is a human subject suffering from or at risk of suffering from any form and / or mutation associated with CF. In some embodiments, the subject is a human subject suffering from or at risk of suffering from hypokalemia associated with CF.

[0042] In certain embodiments, the present invention provides a method for treating, improving and / or preventing hyperglycemia associated with CF in a subject, the method comprising administering to the subject a composition comprising a pharmaceutical agent capable of inhibiting SGLT activity. In some embodiments, the subject is a human subject. In some embodiments, the subject is a human subject suffering from or at risk of suffering from: any form and / or mutation associated with CF. In some embodiments, the subject is a human subject suffering from or at risk of suffering from: hyperglycemia associated with CF.

[0043] In certain embodiments, the present invention provides a method for treating, improving and / or preventing dyslipidemia associated with CF in a subject, the method comprising administering to the subject a composition comprising a pharmaceutical agent capable of inhibiting SGLT activity. In some embodiments, the subject is a human subject. In some embodiments, the subject is a human subject suffering from or at risk of suffering from: any form and / or mutation associated with CF. In some embodiments, the subject is a human subject suffering from or at risk of suffering from: dyslipidemia associated with CF.

[0044] In certain embodiments, the present invention provides a method for treating, improving and / or preventing hypoalbuminemia associated with CF in a subject, the method comprising administering to the subject a composition comprising a pharmaceutical agent capable of inhibiting SGLT activity. In some embodiments, the subject is a human subject. In some embodiments, the subject is a human subject suffering from or at risk of suffering from any form and / or mutation associated with CF. In some embodiments, the subject is a human subject suffering from or at risk of suffering from hypoalbuminemia associated with CF.

[0045] In certain embodiments, the present invention provides a method for treating, improving and / or preventing hypoproteinemia associated with CF in a subject, the method comprising administering to the subject a composition comprising a pharmaceutical agent capable of inhibiting SGLT activity. In some embodiments, the subject is a human subject. In some embodiments, the subject is a human subject suffering from or at risk of suffering from: any form and / or mutation associated with CF. In some embodiments, the subject is a human subject suffering from or at risk of suffering from: hypoproteinemia associated with CF.

[0046] Such methods are not limited to treating specific forms or mutations associated with CF. In some embodiments, the mutation is any mutation associated with a class 1A CTFR mutation (e.g., Dele2,3 (21kb) and 1717-1G→A). In some embodiments, the mutation is any mutation associated with a class 1B CTFR mutation (e.g., Gly542X and Trp1282X). In some embodiments, the mutation is any mutation associated with a class 2 CTFR mutation (e.g., Phe508del, Asn1303Lys, and Ala561Glu). In some embodiments, the mutation is any mutation associated with a class 3 CTFR mutation (e.g., Gly551Asp, Ser549Arg, and Gly1349Asp). In some embodiments, the mutation is any mutation associated with a class 4 CTFR mutation (e.g., Arg117His, Arg334Trp, and Ala455Glu). In some embodiments, the mutation is any mutation associated with a Class 5 CTFR mutation (e.g., 3272-26A→G, 3849+10kg C→T). In some embodiments, the mutation is any mutation associated with a Class 6 CTFR mutation (e.g., c.120del123 and rPhe580del).

[0047] The present invention is not limited to a particular type or class of pharmaceutical agents that function as inhibitors of SGLT activity. In some embodiments, the pharmaceutical agent capable of inhibiting SGLT activity is a small molecule, an antibody, a nucleic acid molecule (eg, siRNA, antisense oligonucleotide), or a peptidomimetic.

[0048] In some embodiments, the pharmaceutical agent capable of inhibiting SGLT activity is selected from, for example, phlorizin, canagliflozin ((2S,3R,4R,5S,6R)-2-{3-[5-[4-fluoro-phenyl)-thiophen-2-ylmethyl]-4-methyl-phenyl}-6-hydroxymethyl-tetrahydro-pyran-3,4,5-triol), dapagliflozin ((2S,3R,4R,5S,6R)-2-[4-chloro-3-(4-ethoxybenzyl)phenyl]-6-(hydroxymethyl)-tetrahydro-2H-pyran-3,4,5-triol), empagliflozin ((2S,3R,4R,5S,6R)-2-[4-chloro-3-[[4-[(3S)-oxolan-3-yl]oxyphenyl]methyl]phenyl]-6-(hydroxymethyl)oxane -3,4,5-triol), repagliflozin (5-methyl-4-[4-(1-methylethoxy)benzyl]-1-(1-methylethyl)-1H-pyrazol-3-yl 6-O-(ethoxycarbonyl)-β-D-pyranoglucopyranoside), sergliflozin (2-(4-methoxybenzyl)phenyl 6-O-(ethoxycarbonyl)-β-D-pyranoglucopyranoside) and togliflozin ((1S,3'R,4'S,5'S,6'R)-6-(4-ethylbenzyl)-6'-(hydroxymethyl)-3',4',5',6'-tetrahydro-3H-spiro[2-benzofuran-1,2'-pyran]-3',4',5'-triol hydrate (1:1)), and sogliflozin (LX4211), or a pharmaceutically acceptable salt thereof.

[0049] An important aspect of the invention is that the compositions of the invention (e.g., compositions comprising a pharmaceutical agent that functions as an inhibitor of SGLT activity) can be used to treat CF and symptoms associated with CF (e.g., hypokalemia, hyperglycemia, dyslipidemia, hypoalbuminemia, and hypoproteinemia).

[0050] Some embodiments of the invention provide methods for administering an effective amount of a composition comprising a pharmaceutical agent that acts as an inhibitor of SGLT activity of the invention and at least one additional therapeutic agent (including, but not limited to, any pharmaceutical agent that can be used to treat CF and / or symptoms associated with CF (e.g., hypokalemia, hyperglycemia, dyslipidemia, hypoalbuminemia, and hypoproteinemia)).

[0051] Compositions within the scope of the present invention include all compositions, wherein the pharmaceutical agent that acts as an SGLT activity inhibitor is included in an amount that effectively achieves its intended purpose. Although individual needs are different, it is within the technical scope of this art to determine the optimal range of the effective amount of each component. Typically, a pharmaceutical agent (e.g., small molecules, antibodies, mimetic peptides) or an equivalent amount of a pharmaceutically acceptable salt thereof that acts as an SGLT activity inhibitor can be orally administered to a mammal (e.g., a human) at a dosage of 0.0025 to 50 mg / kg of mammal body weight per day, and the mammal is receiving treatment for an illness that responds to suppressing SGLT activity. In one embodiment, oral administration of about 0.01 to about 25 mg / kg is used to treat, improve or prevent such illnesses. For intramuscular injection, the dosage is generally about one-half of an oral dose. For example, a suitable intramuscular dose will be about 0.0025 to about 25 mg / kg, or about 0.01 to about 5 mg / kg.

[0052] The unit oral dose can include about 0.01 to about 3000 mg, for example, about 0.1 to about 100 mg of the SGLT activity inhibitor. The unit dose can be administered once or multiple times a day with one or more tablets or capsules, each of which contains about 0.1 to about 10 mg, conveniently about 0.25 to 50 mg of the SGLT activity inhibitor (e.g., peptidomimetics, small molecules) or its solvate.

[0053] In preparations (e.g., intravenous preparations, intraperitoneal preparations, intramuscular preparations, subcutaneous preparations, injection preparations, external preparations, oral preparations, etc.), SGLT activity inhibitors (e.g., mimetic peptides, small molecules) can be present at a concentration of about 0.01 to 100 mg per gram of carrier. In one embodiment, SGLT activity inhibitors (e.g., mimetic peptides, small molecules) are present at a concentration of about 0.07-1.0 mg / ml, such as about 0.1-0.5 mg / ml, and in one embodiment, at a concentration of about 0.4 mg / ml.

[0054] In addition to administering SGLT activity inhibitors (e.g., peptidomimetics, small molecules) as raw chemicals, the SGLT activity inhibitors of the present invention (e.g., peptidomimetics, small molecules) can be administered as part of a pharmaceutical preparation containing a suitable pharmaceutically acceptable carrier, which includes excipients and adjuvants that facilitate the processing of the SGLT activity inhibitor into a pharmaceutically usable preparation. Preparations, particularly those that can be administered in any desired manner (e.g., intravenous, intraperitoneal, intramuscular, subcutaneous, topical, oral, etc.) and can be used for administration in one type of administration, such as tablets, dragees, sustained-release tablets and capsules, gargles and mouthwashes, gels, liquid suspensions, hair dyes, hair gels, shampoos, and preparations that can be administered rectally (such as suppositories), and suitable solutions for intravenous infusion, injection, topical or oral administration, contain about 0.01 to 99 percent, in one embodiment about 0.25 to 75 percent of one or more active peptidomimetics, together with excipients.

[0055] The pharmaceutical composition of the present invention can be administered to any patient who may experience the beneficial effects of the SGLT activity inhibitor of the present invention (e.g., mimetic peptides, small molecules). The most important of such patients are mammals, such as humans, although the present invention is not intended to be limited thereto. Other patients include veterinary animals (cattle, sheep, pigs, horses, dogs, cats, etc.).

[0056] SGLT activity inhibitors (e.g., peptidomimetics, small molecules) and pharmaceutical compositions thereof can be administered in any manner to achieve their intended purpose. For example, administration can be by parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, buccal, intrathecal, intracranial, intranasal or topical routes. Alternatively, or in parallel, administration can be by oral route. The dosage administered will depend on the recipient's age, health and weight, the type of concurrent treatment (if any), the frequency of treatment and the nature of the desired effect.

[0057] The pharmaceutical preparations of the invention are produced in a manner known per se, for example, by means of conventional mixing, granulation, dragee making, dissolution or lyophilization processes. Thus, pharmaceutical preparations for oral use can be obtained by combining the active peptidomimetic with a solid excipient, optionally grinding the resulting mixture and processing the granular mixture, after adding suitable auxiliaries if necessary or necessary to obtain tablets or dragee cores.

[0058] Suitable excipients are especially fillers such as sugars such as lactose or sucrose, mannitol or sorbitol, cellulose preparations and / or calcium phosphates such as tricalcium phosphate or calcium hydrogen phosphate, and binding agents such as starch pastes, using for example corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methylcellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose and / or polyvinyl pyrrolidone. If necessary, disintegrants can be added, such as starch mentioned above and also carboxymethyl starch, cross-linked polyvinyl pyrrolidone, agar or alginic acid or its salt such as sodium alginate. Auxiliary agents are first flow regulators and lubricants, for example, silicon dioxide, talcum, stearic acid or its salt such as magnesium stearate or calcium stearate, and / or polyethylene glycol. The dragee core has a suitable coating, which can resist gastric juice if necessary. For this purpose, concentrated sugar solutions can be used, which can optionally contain gum arabic, talcum, polyvinyl pyrrolidone, polyethylene glycol and / or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures. In order to produce a coating resistant to gastric juice, a solution of a suitable cellulose preparation (such as acetylcellulose phthalate or hydroxypropylmethylcellulose phthalate) is used. Coloring agents or pigments can be added to tablets or dragee coatings, for example, for identification or in order to characterize the combination of active mimetic peptide dosages.

[0059] Other pharmaceutical preparations that can be used orally include push-fit capsules made of gelatin, and soft sealed capsules made of gelatin and a plasticizer (e.g., glycerol or sorbitol). Push-fit capsules can contain the active mimetic peptide in the form of particles, which can be mixed with a filler (such as lactose), a binder (such as starch) and / or a lubricant (such as talc or magnesium stearate) and an optional stabilizer. In a soft capsule, in one embodiment, the active mimetic peptide is dissolved or suspended in a suitable liquid such as a fatty oil or liquid paraffin. In addition, a stabilizer can be added.

[0060] Possible pharmaceutical preparations that can be used for rectal administration include, for example, suppositories consisting of a combination of one or more active mimetic peptides and a suppository base. Suitable suppository bases are, for example, natural or synthetic triglycerides or paraffins. In addition, gelatin rectal capsules consisting of a combination of active mimetic peptides and a base can also be used. Possible base materials include, for example, liquid triglycerides, polyethylene glycols or paraffins.

[0061] Suitable formulations for parenteral administration include aqueous solutions of active mimetic peptides in water-soluble form, such as water-soluble salts and alkaline solutions. In addition, the suspension of the active mimetic peptide as a suitable oily injection suspension can be administered. Suitable lipophilic solvents or carriers include fatty oils, such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides or polyethylene glycol-400. Aqueous injection suspensions can include materials that increase the viscosity of the suspension, including, for example, sodium carboxymethylcellulose, sorbitol and / or dextran. Optionally, the suspension can also include a stabilizer.

[0062] In one embodiment, the topical composition of the present invention is formulated into oil, cream, lotion, ointment, etc. by selecting an appropriate carrier. Suitable carriers include vegetable oil or mineral oil, white petrolatum (white soft paraffin), branched fats or oils, animal fats and high molecular weight alcohols (greater than C 12 ). The carrier may be one in which the active ingredient is soluble. If necessary, an emulsifier, a stabilizer, a humectant and an antioxidant as well as an agent that imparts color or fragrance may also be included. In addition, percutaneous permeation enhancers may be used in these topical preparations. Examples of such enhancers may be found in U.S. Pat. Nos. 3,989,816 and 4,444,762.

[0063] Ointments can be prepared by mixing a solution of the active ingredient in a vegetable oil such as almond oil with warm soft paraffin and allowing the mixture to cool. A typical example of such an ointment is an ointment comprising about 30% almond oil and about 70% white soft paraffin by weight. Lotions can be conveniently prepared by dissolving the active ingredient in a suitable high molecular weight alcohol such as propylene glycol or polyethylene glycol.

[0064] Those of ordinary skill in the art will readily recognize that the foregoing represents only a detailed description of certain preferred embodiments of the invention. Various modifications and variations of the above-described compositions and methods may be readily accomplished using expertise available in the art and are within the scope of the invention.

[0065] Now the present invention has been fully described, it will be appreciated by those skilled in the art that the same can be carried out in a wide and equivalent range of conditions, formulations and other parameters without affecting the scope of the present invention or any embodiment thereof. All patents, patent applications and publications cited herein are incorporated herein by reference in their entirety.

[0066] experiment

[0067] Example I.

[0068] This example demonstrates the beneficial effects of the SGLT inhibitor LX4211 in the treatment of CF.

[0069] To determine whether the dual SGLT1 / 2 inhibitor LX4211

[0070] To determine whether it has a beneficial effect on the treatment of CF, LX4211 (15 mg / kg / day) was administered orally to 5 CF rabbits daily for 4 weeks. Intravenous glucose tolerance test (IVGTT) and insulin tolerance test (ITT) were performed before and 4 weeks after treatment. Urine was collected daily to assess urinary glucose excretion. Blood was collected every two weeks for analysis of chemical panels, including glucose, insulin, liver function, kidney function, lipid profiles, and electrolytes. At the end of the experiment, organ tissues of CF rabbits were collected for histological staining.

[0071] All five CF rabbits showed no obvious adverse reactions to LX4211 treatment. As expected, urine glucose levels rose sharply starting D1 after treatment. Daily abdominal palpation of all animals showed abdominal tenderness. Since most CF rabbits eventually die of intestinal obstruction, abdominal hardening (which can be felt by palpation) has been used as its clinical sign, so this softening may indicate a beneficial effect on relieving GI obstruction, although further studies are needed to confirm this prediction.

[0072] Figure 1 LX4211 is shown to restore glucose tolerance in CF rabbits. One CF rabbit showed clear signs of CF-related diabetes (CFRD) before LX4211 treatment (red line), with the GTT reaction returning to normal after treatment (green line). This result indicates that SGLT inhibitors bring clinical benefits to CFRD. Figure 2 LX4211 treatment was also shown to improve glucose tolerance in CF rabbits. Figure 3 It was shown that LX4211 treatment resulted in improved survival length in CF rabbits.

[0073] LX4211 was shown to provide beneficial effects on electrolyte imbalance and lipid metabolism disorders in CF rabbits. Figure 4 It is shown in Figure 2 that CF rabbits exhibited many abnormalities in metabolic parameters compared to WT, including lower serum potassium, higher triglycerides, cholesterol, and glucose. Abnormalities were also detected in serum ALP, CPK, and calcium ions in CF rabbits. LX4211 treatment significantly alleviated hypokalemia and hyperglycemia in CF rabbits. In addition, significant rescue effects on triglycerides, cholesterol, CPK, and calcium were observed in CF animals after the start of LX4211. These results indicate the beneficial effects of the SGLT2 dual inhibitor LX4211 on electrolyte imbalance and glucose and lipid metabolism disorders in the CF rabbit model. Figure 5 It was shown that LX4211 treatment significantly alleviated hypokalemia and hyperglycemia in CF rabbits. Figure 6 LX4211 is shown to restore total Chol, CPK, ALB and TPRO.

[0074] Example II.

[0075] This example demonstrates that SGLT1 is upregulated in CF-related tissues in CF rabbits.

[0076] First, SGLT1 and SGLT2 transcript (i.e., mRNA) levels were determined in CF and WT rabbits. SGLT2 expression was similar to that reported in humans and other animals, was primarily confined to the kidney, and did not differ between CF and WT rabbits. Interestingly, SGLT1 mRNA levels were elevated in several CF-related tissues, including the trachea, intestine, and liver ( Figure 7 Western blot shows intestine, pancreas ( Figure 7 e) and lungs ( Figure 7 f) The protein level of SGLT1 in tissues of CF rabbits is higher than that of WT rabbits. Consistent immunohistostaining confirmed that the upregulation of SGLT1 in the intestine and pancreas of CF rabbits is higher than that of WT rabbits. In summary, these data show that SGLT1 is upregulated in CF-related tissues of CF rabbits.

[0077] Example III.

[0078] This example demonstrates that SGLT1 is upregulated in CF patient-derived cells.

[0079] Experiments to examine whether SGLT1 expression is also regulated in airway lineage cells derived from human patients were performed. Experiments to examine SGLT1 protein levels in CF bronchial epithelial (CFBE) cells and in CF lung organoids derived from CF patient-specific iPSCs were performed, as previously reported (see, J. Ruan et al., Mol Ther Nucleic Acids https: / / doi.org / 10.1016 / j.omtn.2019.02.006 (2019)).

[0080] The CFTR bands in CFBE cells are consistent with their genotype. The SGLT1 signal is negatively correlated with the CFTR signal: high in CFBE-dF cells, but low in CFBE-WT cells ( Figure 8 , left panel). Consistently, SGLT1 levels were significantly higher in both dF / dF and dF / G551D lung organoids than in WT / WT organoids ( Figure 8 , right).

[0081] These data show that SGLT1 is upregulated in CF patient-derived airway lineage cells as observed in CF rabbits. Taken together, these data suggest that SGLT1 indicates a therapeutic target in CF.

[0082] Example IV.

[0083] This example demonstrates that Sota (LX4211) improves glucose tolerance in CF rabbits.

[0084] The finding that SGLT1 is elevated in several CF-related tissues strongly suggests testing SGLT1 inhibitor drugs in CF rabbits.

[0085] The initial hypothesis was that SGLT1 inhibition might alleviate CFRD symptoms. However, due to the lack of SGLT1 inhibitors at the time, the dual inhibitor Sota, which inhibits both SGLT2 and SGLT1, was used.

[0086] Experiments were performed to first examine the physiological response of rabbits to Sota by measuring urine sugar levels to assess urinary glucose excretion ( Figure 9 ). Sota was administered daily to WT rabbits (n=2) for five days. A sudden rise in urine sugar levels was observed on the second day (D1) and remained high during the treatment days (D1-D5). After the drug was withdrawn, urine sugar levels returned to normal immediately (D7). This confirms that rabbits respond similarly to rodents and human patients to Sota.

[0087] The following treatment experiments were performed by treating CF rabbits (n=6) with Sota (15 mg / kg / day) daily by gavage for 4 weeks ( Figure 10 IVGTT tests were performed before and 4 weeks after treatment. Animals treated with Sota showed a significantly higher rate of blood glucose elimination than the untreated group ( Figure 10 B and C), indicating the beneficial effects of Sota on glucose metabolism in CF rabbits.

[0088] Example V

[0089] This example demonstrates that Sota improves blood chemistry parameters in CF rabbits.

[0090] It is known that many CF patients show abnormalities in their blood chemistry tests. Blood chemistry in CF rabbits was also examined. In one study, blood was collected from 5 CF rabbits for analysis of a chemistry panel, including electrolytes, glucose, and other metabolic parameters, and compared to a chemistry panel of WT rabbits (n=15). CF rabbits showed many abnormalities in metabolic parameters, including lower serum potassium (WT 4.35±0.25 vs. CF 3.14±0.3, p<0.05), higher triglycerides (WT 75.8±69.5 vs. CF447.0±76.6, p<0.05), cholesterol (WT 31.6±9.76 vs. CF 177.2±206.3, p<0.05), and glucose (WT110.4±9.99 vs. CF 160.6±37.5, p<0.05). Serum ALP, CPK, and calcium ions were also detected as abnormal in CF rabbits.

[0091] To evaluate whether Sota has any effect on blood chemistry parameters of CF rabbits, an experiment was performed in which 5 CF rabbits were treated with Sota for 10 weeks. CF rabbits in the control group (n=5) did not receive any Sota treatment.

[0092] Sota treatment significantly improved imbalanced / abnormal parameters such as K+, Trig, Glucose, Chol, ALP and CPK ( Figure 11 In control animals, these parameters gradually deteriorated, whereas in Sota-treated animals, these parameters generally returned to normal ranges ( Figure 7 ). These unexpected findings indicate that the benefits of Sota treatment in CF rabbits extend beyond glucose metabolism and are particularly evident in terms of electrolyte imbalance and lipid metabolism.

[0093] Example VI.

[0094] This example demonstrates that Sota does not affect body weight gain and prolongs the lifespan of CF rabbits.

[0095] A surprising finding of the experiment was that Sota did not affect weight gain, and Sota significantly extended the lifespan of CF rabbits.

[0096] Sota-treated CF rabbits gained weight at a rate similar to that of animals not receiving the drug ( Figure 12 , left), indicating that Sota treatment did not seriously affect the nutritional intake of CF rabbits.

[0097] This is further supported by the extended lifespan of CF rabbits treated with Sota. CF rabbits without Sota (control group, n=10) had a median lifespan of ~60 days, similar to our earlier findings (addition of liquid diet supplement). With Sota treatment (starting at 49 days of age), CF rabbits (n=10) had a median lifespan of >150 days ( Figure 12 , right), and the number of Sota-treated CF rabbits (n=6) surviving more than 150 days was significantly higher than that of the control group (n=1). These findings indicate that, contrary to what many people believe, SGLT inhibitor drugs are indicated to bring benefits with minimal risks to CF patients.

[0098] Figure 13 A summary schematic diagram is provided showing that SGLT1 is upregulated in human CF airway lineage cells and many CF-related tissues in CF rabbits; and the SGLT inhibitor Sota brings about many beneficial effects on CF rabbits.

[0099] Taken together, these data show that SGLT1 is upregulated in CF-related tissues in CF rabbits.

[0100] Equivalent

[0101] The present invention may be embodied in other specific forms without departing from its spirit or essential characteristics. Therefore, the foregoing embodiments are considered to be illustrative in all respects and not limiting the present invention described herein. Therefore, the scope of the present invention is indicated by the appended claims rather than the above description, and all changes within the equivalent meaning and scope of the claims should be included therein.

[0102] Join by reference

[0103] The entire disclosure of each patent document and scientific literature mentioned herein is incorporated by reference for all purposes.

Claims

1. A composition, the composition comprising a pharmaceutical agent capable of inhibiting SGLT activity.

2. A method for inhibiting SGLT activity in a subject, the method comprising administering to the subject the composition according to claim 1.

3. A method for treating, ameliorating, and / or preventing CF in a subject, the method comprising administering to the subject the composition according to claim 1.

4. A method for treating, ameliorating, and / or preventing one or more symptoms associated with CF in a subject, the method comprising administering to the subject the composition according to claim 1.

5. A method for treating, ameliorating, and / or preventing hypokalemia in a subject, the method comprising administering to the subject the composition according to claim 1.

6. A method for treating, ameliorating, and / or preventing hypokalemia in a subject, the method comprising administering to the subject the composition according to claim 1.

7. A method for treating, ameliorating, and / or preventing hyperglycemia in a subject, the method comprising administering to the subject the composition according to claim 1.

8. A method for treating, ameliorating, and / or preventing dyslipidemia in a subject, the method comprising administering to the subject the composition according to claim 1.

9. A method for treating, ameliorating, and / or preventing hypoproteinemia in a subject, the method comprising administering to the subject the composition according to claim 1.

10. A kit, the kit comprising (1) the composition according to claim 1, (2) a container, package, or dispenser, and (3) instructions for administration.

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