Pharmaceutical combinations comprising a cholane derivative and a statin or ursodeoxycholic acid

Abstract

The present invention relates to a pharmaceutical combination comprising 6α-ethyl-3α,7α-dihydroxy-24-nor-5β-cholan-23-ol or a pharmaceutically acceptable salt thereof and a statin, and a combination comprising 6α-ethyl-3α,7α-dihydroxy-24-nor-5β-cholan-23-ol and ursodeoxycholic acid, and uses thereof, for treating non-alcoholic hepatic steatosis and non-alcoholic steatohepatitis.

Description

[Technical Field]

[0001] This patent application claims priority to Italian Patent Application No. 102022000018669, filed September 13, 2022, the entire disclosure of which is incorporated herein by reference.

[0002] The present invention relates to a pharmaceutical composition comprising 6α-ethyl-3α,7α-dihydroxy-24-nor-5β-cholan-23-ol (BAR502). [Background technology]

[0003] Nonalcoholic fatty liver disease (NAFLD) represents the hepatic manifestation of metabolic syndrome and is associated with metabolic abnormalities such as obesity, insulin resistance, fasting hyperglycemia, dyslipidemia, and altered adipokine profiles. Its global prevalence continues to increase with the obesity epidemic, and it has become the most common cause of chronic liver disease in the past decade.

[0004] NAFLD is characterized by excessive lipid accumulation in hepatocytes: in the early stages of the disease, there is simple hepatic steatosis, which can progress to nonalcoholic steatohepatitis (NASH), and in more severe cases, liver fibrosis and cirrhosis can also occur, resulting in an increased risk of developing hepatocellular carcinoma (HCC).

[0005] BAR502 has the following formula:

[0006] [ka]

[0007] It has.

[0008] From WO2015181275 it is known that this is a dual TGR5 / GPBAR1 agonist used in the treatment of NAFLD.

[0009] NAFLD and NASH are highly prevalent syndromes, yet there are no approved effective treatments currently available, resulting in significant interest in the scientific community in identifying new treatments. [Prior art documents] [Patent documents]

[0010] [Patent Document 1] WO2015181275 [Non-patent literature]

[0011] [Non-Patent Document 1] Remington's Pharmaceutical Sciences, 17th ed., Gennaro et al., Mack Publishing Co., 1985 [Non-patent document 2] Remington's Pharmaceutical Sciences, edited by Gennaro AR, 20th edition, 2000, Williams & Wilkins, PA, USA. [Non-patent document 3] Remington: The Science and Practice of Pharmacy, 21st ed., Lippincott Williams & Wilkins, 2005 [Non-patent document 4] Loyd V. Allen and Howard C. Ansel, Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems, 10th edition, edited by Lippincott Williams & Wilkins, 2014. Summary of the Invention

[0012] Therefore, it is an object of the present invention to provide a new treatment for NAFLD and NASH.

[0013] This object is achieved by a pharmaceutical combination according to claim 1, a use thereof according to claim 5, a combination according to claim 7 and a use thereof according to claim 8. [Brief explanation of the drawings]

[0014] The present invention will now be described in detail with reference to the drawings of the accompanying drawings.

[0015] [Figure 1] FIG. 1 shows the change in body weight over time in C57BL6 mice after oral administration of HFD-F alone, HFD-F + BAR502 30 mg / kg, HFD-F + atorvastatin 50 mg / kg, or HFD-F and the combination BAR502 (30 mg / kg) + atorvastatin (50 mg / kg). [Figure 2] FIG. 1 shows the time course of glucose in C57BL6 mice after oral administration of HFD-F alone, HFD-F + BAR502 30 mg / kg, HFD-F + atorvastatin 50 mg / kg, or HFD-F and the combination BAR502 (30 mg / kg) + atorvastatin (50 mg / kg). [Figure 3A] FIG. 1 shows AST levels in the blood of C57BL6 mice measured after oral administration of HFD-F alone, HFD-F + BAR502 30 mg / kg, HFD-F + atorvastatin 50 mg / kg, or HFD-F and the combination BAR502 (30 mg / kg) + atorvastatin (50 mg / kg). [Figure 3B] FIG. 1 shows ALT levels in the blood of C57BL6 mice measured after oral administration of HFD-F alone, HFD-F + BAR502 30 mg / kg, HFD-F + atorvastatin 50 mg / kg, or HFD-F and the combination BAR502 (30 mg / kg) + atorvastatin (50 mg / kg). [Figure 4] FIG. 1 shows blood cholesterol levels in C57BL6 mice after oral administration of HFD-F alone, HFD-F + BAR502 30 mg / kg, HFD-F + atorvastatin 50 mg / kg, or HFD-F and the combination BAR502 (30 mg / kg) + atorvastatin (50 mg / kg). [Figure 5A] FIG. 1 shows histological sections of NT liver. [Figure 5B] FIG. 1 shows histological sections of the liver after oral administration of HFD-F alone. [Figure 5C] FIG. 1 shows histological sections of the liver after oral administration of HFD-F+BAR502 30 mg / kg. [Figure 5D] FIG. 1 shows histological sections of the liver after oral administration of HFD-F plus atorvastatin 50 mg / kg. [Figure 5E] FIG. 1 shows histological sections of the liver after oral administration of the combination of BAR502 (30 mg / kg) plus atorvastatin (50 mg / kg). [Figure 6] FIG. 6 shows the liver steatosis scores in histological sections of the liver referred to in FIG. 5. [Figure 7] FIG. 6 shows the score for hepatic “ballooning” (balloon-like degeneration) in histological sections of the liver referred to in FIG. 5. [Figure 8] FIG. 1 shows the time course of body weight changes in C57BL6 mice after oral administration of HFD-F alone, HFD-F + BAR502 30 mg / kg, HFD-F + UDCA 30 mg / kg, or HFD-F and a combination of BAR502 (30 mg / kg) and UDCA (30 mg / kg). [Figure 9] FIG. 1 shows the time course of blood glucose in C57BL6 mice after oral administration of HFD-F alone, HFD-F + BAR502 30 mg / kg, HFD-F + UDCA 30 mg / kg, or HFD-F and a combination of BAR502 (30 mg / kg) and UDCA (30 mg / kg). [Figure 10] FIG. 1 shows a) AST levels and b) ALT levels in the blood of C57BL6 mice measured after oral administration of HFD-F alone, HFD-F + BAR502 30 mg / kg, HFD-F + UDCA 30 mg / kg, or HFD-F and a combination of BAR502 (30 mg / kg) and UDCA (30 mg / kg). [Figure 11]FIG. 1 shows blood cholesterol levels in C57BL6 mice after oral administration of HFD-F alone, HFD-F + BAR502 30 mg / kg, HFD-F + UDCA 30 mg / kg, or HFD-F and a combination of BAR502 (30 mg / kg) and UDCA (30 mg / kg). [Figure 12] FIG. 1 shows histological analysis of liver sections after oral administration of HFD-F alone, HFD-F + BAR502 30 mg / kg, HFD-F + UDCA 30 mg / kg, or HFD-F and a combination of BAR502 (30 mg / kg) and UDCA (30 mg / kg). [Figure 13A] FIG. 1 shows the scores related to ballooning (lipid deposition) in the different treatment groups. [Figure 13B] FIG. 1 shows scores related to the severity of steatosis in different treatment groups. DETAILED DESCRIPTION OF THE INVENTION

[0016] According to a first aspect of the present invention, there is provided a pharmaceutical combination comprising 6α-ethyl-3α,7α-dihydroxy-24-nor-5β-cholan-23-ol or a pharmaceutically acceptable salt thereof and a statin.

[0017] In one embodiment, the statin is selected from the group consisting of atorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin, pravastatin, rosuvastatin and simvastatin, preferably it is atorvastatin.

[0018] According to another aspect of the present invention, there is provided a pharmaceutical combination comprising 6α-ethyl-3α,7α-dihydroxy-24-nor-5β-cholan-23-ol or a pharmaceutically acceptable salt thereof and ursodeoxycholic acid or a pharmaceutically acceptable salt thereof.

[0019] The combinations according to the first and second aspects of the invention may further comprise at least one pharmaceutically acceptable excipient.

[0020] The combinations of the present invention may be included in pharmaceutical compositions and dosage units thereof, in such form they can be used as solids such as filled tablets or capsules, or as liquids such as solutions, suspensions, emulsions, elixirs, or as filled capsules thereof, or in the form of sterile injectable solutions for parenteral administration (including subcutaneous and intravenous use), all for oral use.

[0021] Such pharmaceutical compositions and unit dosage forms thereof may contain the ingredients in conventional proportions, with or without additional compounds or active ingredients, and such unit dosage forms may contain any suitable effective amount of each active ingredient corresponding to the predetermined daily dosage interval being used.

[0022] The pharmaceutical composition containing the combination of the present invention can be prepared by methods well known in the pharmaceutical art.Generally, the combination of the present invention is administered in a pharmaceutically effective amount.The amount of the combination actually administered will generally be determined by a physician in consideration of the relevant circumstances, including the condition to be treated, the selected administration route, the actual combination to be administered, the age, weight and response of each patient, the severity of the patient's symptoms, etc.

[0023] Pharmaceutical compositions containing the combination of the present invention can be administered by several routes, including oral, rectal, subcutaneous, intravenous, intramuscular, intranasal, and pulmonary routes. Compositions for oral administration can take the form of bulk solutions or suspensions or bulk powders. More commonly, however, compositions are provided in unit dosage forms to facilitate accurate dosing. The term "unit dosage form" refers to physically discrete units suitable as unit dosages for human and other mammalian subjects, each containing a predetermined amount of active substance calculated to produce the desired therapeutic effect, together with acceptable pharmaceutical excipients. Typical unit dosage forms include pre-filled, pre-dosed ampoules or syringes of liquid compositions, or pills, tablets, capsules, or the like in the case of solid compositions.

[0024] Liquid forms suitable for oral administration may include a suitable aqueous or nonaqueous vehicle with buffers, suspending and dispersing agents, dyes, flavors, etc. Solid forms may include, for example, any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth, or gelatin; an excipient such as starch or lactose; a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate; a flow agent such as colloidal silicon dioxide; a sweetener such as sucrose, lactose, or saccharin; or a flavoring such as peppermint, methyl salicylate, or orange flavoring.

[0025] Injectable compositions are generally based upon sterile injectable solution or phosphate buffer solution or other injectable vehicles known in the art.

[0026] The pharmaceutical compositions may be in the form of tablets, pills, capsules, solutions, suspensions, emulsions, powders, suppositories, and as sustained release formulations.

[0027] If desired, tablets can be coated using standard aqueous or nonaqueous techniques. In some embodiments, such compositions and preparations may contain at least 0.1% of the active compound. The percentage of the active compound in these compositions may, of course, vary and may suitably be from about 1% to about 60% of the unit mass. The amount of active compound in such therapeutically useful compositions is such that a therapeutically active dosage is obtained. The active compound may also be administered intranasally, for example, as drops or spray.

[0028] Tablets, pills, capsules, etc. may also contain binders such as tragacanth gum, gum arabic, cornstarch, or gelatin; excipients such as dicalcium phosphate; disintegrating agents such as cornstarch, potato starch, or alginic acid; lubricants such as magnesium stearate; and sweeteners such as sucrose, lactose, or saccharin. When the dosage unit is a capsule, in addition to the above-mentioned materials, it may contain a liquid carrier such as a fatty oil. Various other materials may be present as coatings or to modify the physical form of the dosage unit. For example, tablets may be coated with shellac, sugar, or both. A syrup or elixir may contain, in addition to the active ingredient, sucrose as a sweetener, methylparaben and propylparaben as preservatives, a dye, and a flavoring such as cherry or orange flavoring. To prevent breakage during passage through the upper gastrointestinal tract, the composition is enterically coated.

[0029] Compositions for pulmonary administration include, but are not limited to, dry powder compositions consisting of a powdered active compound and a powder of a suitable carrier and / or lubricant. Compositions for pulmonary administration can be inhaled from any suitable dry powder inhaler device known to those skilled in the art.

[0030] The composition is administered according to a protocol and at a dosage sufficient to reduce inflammation and pain in the subject. In some embodiments, the active ingredient in the pharmaceutical composition is generally formulated in dosage units. The dosage units may contain 0.1 to 1000 mg of the active compound per daily administration.

[0031] In some embodiments, the effective amount for a particular formulation will depend on the severity of the disease, disorder, or condition, previous treatment, the individual's health status, and response to the drug, hi some embodiments, the dosage ranges from 0.001% to about 60% by weight of the formulation.

[0032] With regard to formulations associated with any type of administration route, methods and formulations for administration of drugs are described in Remington's Pharmaceutical Sciences, 17th Edition, edited by Gennaro et al., Mack Publishing Co., 1985 and Remington's Pharmaceutical Sciences, edited by Gennaro AR, 20th Edition, 2000, Williams & Wilkins PA, USA and Remington: The Science and Practice of Pharmacy, 21st Edition, edited by Lippincott Williams & Wilkins, 2005, and in Lloyd V. Allen and Howard C. Ansel, Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems, 10th Edition, edited by Lippincott Williams & Wilkins, 2014.

[0033] The above ingredients for orally administered or injectable compositions are merely representative.

[0034] The combinations of this invention can also be administered in sustained release forms or by sustained release drug delivery systems.

[0035] Alternatively, the active ingredients of the combination of the present invention are not combined in a single pharmaceutical formulation but may be administered simultaneously or separately at different times, conjointly or independently.

[0036] According to another aspect of the present invention, the above pharmaceutical combination can be used for the treatment of a disorder selected from the group consisting of non-alcoholic hepatic steatosis and non-alcoholic steatohepatitis.

[0037] Description of the embodiment Below, the present invention is illustrated by some examples, which are not intended to limit the scope of the present invention. [Example]

[0038] Example 1. Efficacy of a pharmaceutical combination containing BAR502 and atorvastatin Several preclinical NASH models are available. Among mouse models, steatohepatitis induced by chronic administration of a high-fat (HFD) and fructose (F) diet leads to the development of steatosis, inflammation, and fibrosis, which shows the best correlation with what can be observed in human disease. Using this mouse model, we investigated the efficacy of the combination of BAR502 and atorvastatin in preventing the development of NASH.

[0039] method Twelve-week-old C57BL6 mice were fed a diet containing 60% of calories from fat and fructose (42 g / L) added to the drinking water (HFD-F) or a control diet for 61 days. Mice were randomized to receive HFD-F alone, HFD-F + BAR502 30 mg / kg, HFD-F + atorvastatin 50 mg / kg, or HFD-F and the combination BAR502 (30 mg / kg) + atorvastatin (50 mg / kg) by oral administration starting on day 6.

[0040] result The results obtained in this mouse model surprisingly demonstrate that the combination of BAR502 and atorvastatin exerts a much greater beneficial effect than any single treatment. While all three treatments exhibit beneficial effects on weight gain (Figure 1), the combination of BAR502 and atorvastatin exhibits a significantly greater beneficial effect on reducing insulin resistance than any of the individual treatments, as shown by the glucose tolerance curve (OGTT) results (Figure 2). This data point is of particular interest because insulin resistance represents one of the main hallmarks of human disease. Transaminase analysis (Figures 3a and 3b) showed that all three drug treatments effectively reduced liver damage, but hypercholesterolemia was significantly reduced only by the combination of BAR502 and atorvastatin (Figure 4). This effect exerted by the combination of treatments alone is of great interest because patients with NASH exhibit elevated blood cholesterol levels, which represents one of the main risk factors for the development of vascular disease.

[0041] The main characteristic of NAFLD is lipid deposition at the liver level, which initially leads to steatosis and then steatohepatitis. Histological analysis of this aspect of the disease in the mouse model used (Figures 5A-5E) showed that the HFD-F diet induces high lipid deposition at the liver level, resulting in hepatocyte ballooning and cell death (as evidenced by increased AST and ALT values ​​in this experimental group). Surprisingly, the combination of BAR502 and atorvastatin completely prevented lipid deposition within hepatocytes, thus protecting hepatocytes from ballooning (Figure 7) and the resulting liver damage, as shown by histology and steatosis scores (Figure 6).

[0042] Example 2. Efficacy of a pharmaceutical combination containing BAR502 and ursodeoxycholic acid (UDCA) The mouse model of Example 1 was used again. Therefore, using this mouse model, the inventors investigated the effectiveness of the cooperation between BAR502 and UDCA in preventing the development of NASH.

[0043] method Twelve-week-old C57BL6 mice were fed a diet containing 60% of calories from fat and fructose (42 g / L) in the drinking water (HFD-F) or a control diet for 61 days. Starting on day 6, mice were randomized to receive HFD-F alone, HFD-F + BAR502 30 mg / kg, HFD-F + UDCA 30 mg / kg, or HFD-F and a combination of BAR502 (30 mg / kg) and UDCA (30 mg / kg) by oral administration.

[0044] result The results obtained in this mouse model surprisingly demonstrate that the combination of BAR502 and UDCA exerts a much greater beneficial effect than either single treatment. For example, only the combination of BAR502 and UDCA exhibited a significantly beneficial effect on weight gain (Figure 8), and the glucose tolerance curve (OGTT) results showed that a beneficial effect on insulin resistance was exerted by both the combination and single treatment with UDCA (Figure 9). This data point is of great interest because insulin resistance represents one of the main hallmarks of human disease. Transaminase analysis (Figure 10) showed that all three drug treatments effectively reduced liver damage, whereas hypercholesterolemia (Figure 11) was significantly reduced only by the combination of BAR502 and UDCA. Because patients with NASH exhibit elevated blood cholesterol levels, which represent one of the main risk factors for the development of vascular disease, this effect exerted by the treatment combination alone is of high translational / clinical interest.

[0045] The main characteristic of NAFLD is lipid deposition at the liver level, which initially leads to steatosis and then steatohepatitis. Histological analysis of this aspect of the disease in the mouse model used showed that the HFD-F diet induces high lipid deposition at the liver level, resulting in hepatocyte ballooning and cell death (evidenced by increased AST and ALT values ​​in this experimental group) and liver fibrosis. Surprisingly, the combination of BAR502 and UDCA prevented lipid deposition within hepatocytes with a much greater effect than either treatment exerted individually, as shown by histology (Figure 12) and steatosis scores (Figure 13), thus protecting hepatocytes from ballooning and the resulting liver damage and fibrosis, which represents one of the most dreaded complications of human disease.

Claims

1. A pharmaceutical combination comprising 6α-ethyl-3α,7α-dihydroxy-24-nor-5β-cholan-23-ol or a pharmaceutically acceptable salt thereof and a statin.

2. 2. The pharmaceutical combination according to claim 1, wherein the statin is selected from the group consisting of atorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin, pravastatin, rosuvastatin and simvastatin.

3. 3. The pharmaceutical combination according to claim 2, wherein the statin is atorvastatin.

4. 4. The pharmaceutical combination according to any one of claims 1 to 3, further comprising at least one pharmacologically acceptable excipient.

5. 5. The pharmaceutical combination of any one of claims 1 to 4 for use in the treatment of a disorder selected from the group consisting of non-alcoholic hepatic steatosis and non-alcoholic steatohepatitis.

6. The pharmaceutical combination according to claim 5, wherein the 6α-ethyl-3α,7α-dihydroxy-24-nor-5β-cholan-23-ol or a pharmaceutically acceptable salt thereof and the statin are in the form of a kit-of-parts for combined administration, administered simultaneously or separately at intervals, jointly or independently.

7. A pharmaceutical combination comprising 6α-ethyl-3α,7α-dihydroxy-24-nor-5β-cholan-23-ol or a pharmaceutically acceptable salt thereof and ursodeoxycholic acid or a pharmaceutically acceptable salt thereof.

8. 8. The pharmaceutical combination of claim 7, further comprising at least one pharmacologically acceptable excipient.

9. 9. The pharmaceutical combination according to claim 7 or 8 for use in the treatment of a disorder selected from the group consisting of non-alcoholic hepatic steatosis and non-alcoholic steatohepatitis.

10. The pharmaceutical combination according to claim 9, wherein the 6α-ethyl-3α,7α-dihydroxy-24-nor-5β-cholan-23-ol or a pharmaceutically acceptable salt thereof and the ursodeoxycholic acid or a pharmaceutically acceptable salt thereof are in the form of a kit-of-parts for combined administration, administered simultaneously or separately at intervals, jointly or independently.

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