Pharmaceutical combination comprising cholane derivatives and statins or ursodesoxycholic acid

Abstract

The present invention relates to a pharmaceutical composition comprising 6 [alpha]-ethyl-3 [alpha], 7 [alpha]-dihydroxy-24-nor-5beta-cholane-23-ol or a pharmaceutically acceptable salt thereof and a statin, and to a composition comprising 6 [alpha]-ethyl-3 [alpha], 7 [alpha]-dihydroxy-24-nor-5beta-cholane-23-ol and ursodesoxycholic acid, in particular to a pharmaceutical composition comprising 6 [alpha]-ethyl-3 [alpha], 7 [alpha]-dihydroxy-24-nor-5beta-cholane-23-ol or a pharmaceutically acceptable salt thereof and a statin. And their use for the treatment of non-alcoholic liver steatosis and non-alcoholic steatohepatitis.

Description

[0001] Cross - Reference to Related Applications

[0002] This patent application claims priority to Italian Patent Application No. 102022000018669, filed on September 13, 2022, the entire disclosure of which is incorporated herein by reference. Technical Field

[0003] The present invention relates to a pharmaceutical composition comprising 6α-ethyl-3α,7α-dihydroxy-24-nor-5β-cholan-23-ol (BAR502). Background of the Invention

[0005] Non-alcoholic fatty liver disease (NAFLD) is a manifestation of the metabolic syndrome in the liver, which is associated with metabolic abnormalities such as obesity, insulin resistance, fasting hyperglycemia, dyslipidemia, and adipokine alterations. The prevalence of NAFLD is continuously increasing worldwide, and the increasing obesity has become the most common cause of chronic liver disease in the past decade.

[0006] NAFLD is characterized by the excessive accumulation of lipids in hepatocytes: in the early stage of the disease, it presents as simple hepatic steatosis, which can progress to non-alcoholic steatohepatitis (NASH), and in more severe cases, liver fibrosis and cirrhosis can also occur, thus increasing the risk of developing hepatocellular carcinoma (HCC).

[0007] BAR502 has the following formula:

[0008]

[0009] It is known from WO2015181275 that this compound is a dual agonist of TGR5 / GPBAR1 for the treatment of NAFLD.

[0010] NAFLD and NASH are widespread syndromes, yet there are currently no approved effective therapies, so the scientific community has a strong interest in finding new therapies. Summary of the Invention

[0011] Therefore, an object of the present invention is to provide a new treatment method for NAFLD and NASH.

[0012] This object is achieved by the pharmaceutical combination of claim 1, its use of claim 5, the combination of claim 7, and its use of claim 8.

[0013] Brief Description of the Drawings

[0014] The present invention will now be described in detail with reference to the accompanying drawings, wherein:

[0015] -Figure 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 a combination of HFD-F and BAR502 (30 mg / kg) + atorvastatin (50 mg / kg);

[0016] - Figure 2 Shows the change in glucose 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 a combination of HFD-F and BAR502 (30 mg / kg) + atorvastatin (50 mg / kg);

[0017] -Figure 3 shows the levels of a) AST and b) ALT in the blood measured in C57BL6 mice after oral administration of HFD-F alone, HFD-F + BAR502 30 mg / kg, HFD-F + atorvastatin 50 mg / kg, or a combination of HFD-F and BAR502 (30 mg / kg) + atorvastatin (50 mg / kg);

[0018] - Figure 4 Shows the cholesterol level in the blood of C57BL6 mice after oral administration of HFD-F alone, HFD-F + BAR502 30 mg / kg, HFD-F + atorvastatin 50 mg / kg, or a combination of HFD-F and BAR502 (30 mg / kg) + atorvastatin (50 mg / kg);

[0019] -Figure 5 shows histological sections of the liver after oral administration of HFD-F alone, HFD-F + BAR502 30 mg / kg, HFD-F + atorvastatin 50 mg / kg, or a combination of HFD-F and BAR502 (30 mg / kg) + atorvastatin (50 mg / kg);

[0020] - Figure 6 Shows the score of hepatic steatosis on the liver tissue sections in Figure 5;

[0021] - Figure 7 Shows the score of liver "ballooning" (ballooning degeneration) on the liver tissue sections in Figure 5;

[0022] - Figure 8Shows 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 + UDCA 30 mg / kg, or a combination of HFD-F and BAR502 (30 mg / kg) and UDCA (30 mg / kg);

[0023] - Figure 9 Shows the change in glucose in the blood over time in C57BL6 mice after oral administration of HFD-F alone, HFD-F + BAR502 30 mg / kg, HFD-F + UDCA 30 mg / kg, or a combination of HFD-F and BAR502 (30 mg / kg) and UDCA (30 mg / kg);

[0024] - Figure 10 Shows the levels of a) AST and b) ALT in C57BL6 mice after oral administration of HFD-F alone, HFD-F + BAR502 30 mg / kg, HFD-F + UDCA 30 mg / kg, or a combination of HFD-F and BAR502 (30 mg / kg) and UDCA (30 mg / kg);

[0025] - Figure 11 Shows the cholesterol level in the blood of C57BL6 mice after oral administration of HFD-F alone, HFD-F + BAR502 30 mg / kg, HFD-F + UDCA 30 mg / kg, or a combination of HFD-F and BAR502 (30 mg / kg) and UDCA (30 mg / kg);

[0026] - Figure 12 Shows the 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 a combination of HFD-F and BAR502 (30 mg / kg) and UDCA (30 mg / kg);

[0027] -Figure 13 shows a) the score related to ballooning degeneration (lipid deposition) in each treated group; b) the score related to the severity of steatosis in each treatment group. Detailed Description

[0028] 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.

[0029] In one embodiment, the statins are selected from atorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin, pravastatin, rosuvastatin, and simvastatin, with atorvastatin being preferred.

[0030] 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.

[0031] The combination according to the first and second aspects of the present invention may further comprise at least one pharmacologically acceptable excipient.

[0032] The combination of the present invention may be included in pharmaceutical compositions and their dosage units, and the forms used may be solid (such as filled tablets or capsules), or liquid (such as solutions, suspensions, emulsions, elixirs, or capsules filled with the above substances), all in the form of sterile injectable solutions for oral use or for parenteral administration (including subcutaneous and intravenous use).

[0033] Such pharmaceutical compositions and their unit dosage forms may contain conventional percentages of ingredients, with or without additional compounds or active ingredients, and such unit dosage forms may contain any suitable effective amount of various active ingredients commensurate with the intended daily dose interval used.

[0034] The pharmaceutical composition containing the combination of the present invention can be prepared in a manner well-known in the pharmaceutical art. Generally, the combination of the present invention is administered in a pharmaceutically effective amount. The actual dosage administered is usually determined by a physician based on relevant circumstances such as the condition to be treated, the selected route of administration, the actual combination administered, the age, weight, and response of the individual patient, and the severity of the patient's symptoms.

[0035] The pharmaceutical composition containing the combination of the present invention can be administered by various routes, including oral, rectal, subcutaneous, intravenous, intramuscular, intranasal, and pulmonary routes. The composition for oral administration can be in the form of a bulk liquid solution or suspension or a bulk powder. However, more commonly, the composition exists in unit dosage forms to facilitate precise administration. The term "unit dosage form" refers to physically discrete units suitable as unit doses for human and other mammalian subjects, each unit containing a predetermined amount of the active substance calculated to produce the desired therapeutic effect and an acceptable pharmaceutical excipient. Typical unit dosage forms include pre-filled, pre-measured ampoules or syringes of liquid compositions or pills, tablets, capsules, etc. in the case of solid compositions.

[0036] Liquid forms suitable for oral administration may include suitable aqueous or non-aqueous carriers as well as buffering agents, suspending and dispersing agents, dyes, flavoring agents, and the like. Solid forms may include, for example, any of the following ingredients or compounds of similar nature: binders such as microcrystalline cellulose, tragacanth, or gelatin; excipients such as starch or lactose; disintegrants such as alginic acid, Primogel, or corn starch; lubricants such as magnesium stearate; glidants such as colloidal silicon dioxide; sweetening agents such as sucrose, lactose, or saccharin; or flavoring agents such as peppermint, methyl salicylate, or orange flavoring.

[0037] Injectable compositions are generally based on sterile injectable solutions or phosphate buffered solutions or other injectable carriers known in the art.

[0038] The pharmaceutical composition can be in the form of tablets, pills, capsules, solutions, suspensions, emulsions, powders, suppositories, and sustained release formulations.

[0039] If desired, tablets can be coated by standard aqueous or non-aqueous techniques. In some embodiments, such compositions and formulations can contain at least 0.1% of the active compound. Of course, the percentage of the active compound in these compositions can vary and can suitably be between about 1% and about 60% by unit weight. The amount of the active compound in the therapeutic composition ensures that a therapeutically active dose is obtained. The active compound can also be administered intranasally, such as in the form of droplets or sprays.

[0040] Tablets, pills, capsules, etc. can also contain binders such as tragacanth, gum arabic, corn starch, or gelatin; excipients such as dicalcium phosphate; disintegrants such as corn starch, potato starch, alginic acid; lubricants such as magnesium stearate; and sweetening agents such as sucrose, lactose, or saccharin. When the dosage unit form is a capsule, it can also contain a liquid carrier such as a fatty oil in addition to the materials of the above types. Various other materials can also be present as coatings or for altering the physical form of the dosage unit. For example, tablets can be coated with shellac, sugar, or both. Syrups or elixirs, in addition to containing the active ingredient, can contain sucrose as a sweetening agent, methylparaben and propylparaben as preservatives, dyes, and flavoring agents (such as cherry or orange flavor). To avoid the composition being destroyed during passage through the upper part of the gastrointestinal tract, the composition is an enteric-coated formulation.

[0041] Compositions for pulmonary administration include, but are not limited to, dry powder compositions consisting of powdered active compounds and suitable carriers and / or lubricants. Compositions for pulmonary administration can be inhaled through any suitable dry powder inhaler known to those skilled in the art.

[0042] The administration of the composition is carried out according to a regimen and dose sufficient to alleviate inflammation and pain in a subject. In some embodiments, the active ingredient in the pharmaceutical composition is typically formulated in dosage units. For each dosage unit of the daily dosage, the dosage unit may contain from 0.1 to 1000 mg of the active compound.

[0043] In some embodiments, the effective amount of a particular formulation will depend on the severity of the disease, disorder or condition, previous therapy, the health status of the individual and the response to the drug. In some embodiments, the dosage range is from 0.001 wt% to about 60 wt% of the formulation.

[0044] Regarding formulations for any type of administration route, the methods of drug administration and formulations are described in the following documents: "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 Publishers, Pennsylvania, USA; and "Remington: The Science and Practice of Pharmacy", 21st Edition, Lippincott Williams & Wilkins Publishing, 2005; and "Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems" by Loyd V. Allen and Howard C. Ansel, 10th Edition, Lippincott Williams & Wilkins Publishing, 2014.

[0045] The components for the above oral administration or injectable compositions are merely representative.

[0046] The combination of the present invention can also be administered in a sustained release form or through a sustained release drug delivery system.

[0047] Optionally, although the active ingredients of the combination of the present invention are not formulated as a single pharmaceutical formulation, they can be administered simultaneously in combination or separately or at intervals.

[0048] According to another aspect of the present invention, the above drug combination can be used to treat conditions selected from non-alcoholic hepatic steatosis and non-alcoholic steatohepatitis.

[0049] Description of Embodiments

[0050] Hereinafter, the present invention will be illustrated by some examples, which are not intended to limit the scope of the present invention.

[0051] Example 1. Efficacy of a Pharmaceutical Combination Comprising BAR502 and Atorvastatin

[0052] Several preclinical NASH models can be used. In mouse models, steatohepatitis induced by long-term administration of a high-fat (HFD) and fructose (F) diet that leads to the development of steatosis, inflammation, and fibrosis has shown the best correlation with the outcomes observable in human disease. Using this mouse model, the inventors studied the efficacy of the combination between BAR502 and atorvastatin in preventing the development of NASH.

[0053] Methods

[0054] Twelve-week-old C57BL6 mice were fed a diet containing 60% fat calories with fructose (42 g / l) added to the drinking water (HFD-F) or a control diet for 61 days. Starting from day 6, the mice were randomly administered orally with either only HFD-F, HFD-F + BAR502 (30 mg / kg), HFD-F + atorvastatin (50 mg / kg), or a combination of HFD-F and BAR502 (30 mg / kg) + atorvastatin (50 mg / kg).

[0055] Results

[0056] The results obtained in this mouse model surprisingly showed that the combination of BAR502 and atorvastatin had more significant benefits than single treatment methods. All three treatment methods showed beneficial effects on weight gain ( Figure 1 ), but the combination of BAR502 and atorvastatin showed significantly higher beneficial effects in reducing insulin resistance than single treatment ( Figure 2 ), as shown by the results of the glucose tolerance curve (OGTT). This data item is very interesting because insulin resistance is one of the main features of human diseases. Transaminase analysis (Figures 3a and 3b) indicated that all three drug treatments were effective in reducing liver injury, while hypercholesterolemia was significantly reduced only by the combination of BAR502 and atorvastatin ( Figure 4 ). The effects produced by the combined therapy alone are very interesting because the blood cholesterol concentration of patients with NASH is elevated, which is one of the main risk factors for the development of vascular diseases.

[0057] The main feature of NAFLD is lipid deposition at the liver level, which first leads to steatosis and then to steatohepatitis. Histological analysis of this aspect of the disease in the mouse model used ( Figures 5A - 5E)The HFD-F diet was shown to induce high lipid deposition at the liver level, followed by hepatocyte ballooning degeneration and cell death (as confirmed by the increase in AST and ALT values in this experimental group). Surprisingly, the combination of BAR502 and atorvastatin completely prevented lipid deposition in hepatocytes, as confirmed by histology and steatosis scores( Figure 6 ), thus protecting hepatocytes from ballooning degeneration( Figure 7 ) and avoiding subsequent liver injury.

[0058] Example 2. Efficacy of a pharmaceutical combination comprising BAR502 and ursodeoxycholic acid (UDCA)

[0059] The mouse model of Example 1 was used again. Using this mouse model, the inventors studied the effectiveness of the combination of BAR502 and UDCA in preventing the development of NASH.

[0060] Method

[0061] Twelve-week-old C57BL6 mice were fed a diet containing 60% fat calories and supplemented with fructose (42 g / l) in drinking water (HFD-F) or a control diet for 61 days. Starting from day 6, the mice were randomly administered orally with either HFD-F alone, HFD-F + BAR502 (30 mg / kg), HFD-F + UDCA (30 mg / kg), or a combination of HFD-F and BAR502 (30 mg / kg) with UDCA (30 mg / kg).

[0062] Results

[0063] The results obtained in this mouse model surprisingly showed that the combination of BAR502 and UDCA exerted a more significant beneficial effect than monotherapy. For example, the combination of BAR502 and UDCA alone showed a significant beneficial effect on weight gain( Figure 8 ), however, as shown by the results of the glucose tolerance curve (OGTT), in addition to the combination exerting a beneficial effect on insulin resistance, a beneficial effect on insulin resistance was also exerted by monotherapy with UDCA( Figure 9 ). This data item is very interesting because insulin resistance represents one of the main characteristics of human diseases. Transaminase analysis( Figure 10 ) showed that all three drug treatments were effective in reducing liver injury, while hypercholesterolemia was significantly reduced only by the combination of BAR502 and UDCA( Figure 11 ). This effect produced by the combination therapy alone has high translational / clinical significance because the blood cholesterol concentration of patients with NASH is elevated, which is one of the main risk factors for the development of vascular diseases.

[0064] The main feature of NAFLD is lipid deposition at the liver level, which first leads to steatosis and then to steatohepatitis. Histological analysis of this aspect of the disease in the mouse model used showed that the HFD-F diet induced high lipid deposition at the liver level, followed by hepatocyte ballooning and cell death (as confirmed by the increase in AST and ALT values in this experimental group) and liver fibrosis. Surprisingly, the combination of BAR502 and UDCA had a more significant effect compared to single treatments, preventing lipid deposition in hepatocytes, as confirmed by histology ( Figure 12 ) and steatosis scores (Figure 13), thus protecting hepatocytes from ballooning and avoiding subsequent liver injury and fibrosis, which are among the most dreaded complications of the human disease.

Claims

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

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

4. The pharmaceutical combination according to any one of claims 1 to 3, characterized in that, The pharmaceutical combination further comprises at least one pharmaceutically acceptable excipient.

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

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

7. A drug combination, characterized in that, The pharmaceutical combination comprises 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. The pharmaceutical combination according to claim 7, wherein The pharmaceutical combination further comprises at least one pharmaceutically acceptable excipient.

9. Use of the pharmaceutical combination according to any one of claims 7 or 8 for the treatment of a disorder selected from the group consisting of non-alcoholic hepatic steatosis and non-alcoholic steatohepatitis.

10. The pharmaceutical combination for use according to claim 9, in the form of a multi-component kit for combined administration, 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 administered jointly or simultaneously independently, or separately at intervals.

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