Tetromethamine salts of 4-chloro-5-[4-(2, 6-dichlorophenyl) sulfonylpiperazin-1-yl]-1-benzofuran-2-carboxylic acid and various forms thereof

Abstract

Provided are tromethamine salts of 4-chloro-5-[4-(2, 6-dichlorophenyl) sulfonylpiperazin-1-yl]-1-benzofuran-2-carboxylic acid, and forms thereof, as well as methods for preparing the tromethamine salts. Also provided are pharmaceutical compositions or veterinary compositions comprising the tromethamine salts and their use for the treatment of a number of diseases.

Description

Technical Field

[0001] This invention relates to the field of medicine, and more specifically, to a novel 4-chloro-5-[4-(2,6-dichlorophenyl)sulfonylpiperazin-1-yl]-1-benzofuran-2-carboxylate, its different forms, its preparation methods, and its use in treating a variety of diseases. Background Technology

[0002] 4-Chloro-5-[4-(2,6-dichlorophenyl)sulfonylpiperazin-1-yl]-1-benzofuran-2-carboxylic acid (IUPAC name), CAS number 1192171-69-9, is also currently referred to as "Vonafexor" or "PLX007", and is also referred to as "EYP001" or "EYP-001" in this article. EYP001 has been proven to be a potent FXR agonist, particularly for the treatment of diseases or conditions associated with FXR receptor activity, including hyperlipoproteinemia, hypertriglyceridemia, dyslipidemia, lipodystrophy, cholestasis / fibrosis, cholesterol gallstones, gastrointestinal diseases or conditions, hyperglycemia, diabetes, type 2 diabetes, insulin resistance, metabolic stenosis, kidney disease, liver disease, atherosclerosis, cancer, inflammatory conditions, obesity, osteoporosis, skin aging, hair growth regulation and pigmentation disorders, Parkinson's disease and / or Alzheimer's disease.

[0003] Although the therapeutic effects of EYP001 free acids have been extensively studied, their therapeutic properties have limited their application. To date, research on EYP001 has been limited to its free acid form. Summary of the Invention

[0004] This paper presents a novel EYP001 salt with improved properties. More specifically, it presents an aminobutanetriol salt of 4-chloro-5-[4-(2,6-dichlorophenyl)sulfonylpiperazin-1-yl]-1-benzofuran-2-carboxylic acid (EYP001). This salt exhibits better solubility, particularly in the gastrointestinal tract, compared to, for example, the free acid form of EYP001 and other salts. It also possesses improved pharmacokinetic properties. Therefore, higher bioavailability and lower variability can be achieved relative to the free acid form of EYP001.

[0005] Different forms of EYP001 tromethamine salt, including amorphous and various crystalline forms, have been observed and / or isolated.

[0006] Therefore, this paper provides a methanotrophic salt of 4-chloro-5-[4-(2,6-dichlorophenyl)sulfonylpiperazin-1-yl]-1-benzofuran-2-carboxylic acid (EYP001) in crystalline form.

[0007] In one respect, the crystalline form I of EYP001 tromethamine salt, which exhibits excellent thermal stability (melting point above 180°C) and is non-hygroscopic, has been isolated.

[0008] Therefore, this paper provides a crystalline form (Form I) of an aminobutadiene trioxide of 4-chloro-5-[4-(2,6-dichlorophenyl)sulfonylpiperazin-1-yl]-1-benzofuran-2-carboxylic acid (EYP001), wherein the X-ray diffraction pattern of the crystalline form contains peaks at the following diffraction angles 2-Theta (2θ): 15.2°±0.2°, 16.0°±0.2°, 17.1°±0.2°, 18.3°±0.2°, 18.7°±0.2°, 20.6°±0.2°, 22.6°±0.2°, and 23.2°±0.2°, wherein the X-ray diffraction pattern is obtained using a Cu Kα anode. In a preferred embodiment, the crystalline form I of EYP001 tromethamine salt results in its X-ray diffraction pattern containing peaks at the following diffraction angles 2-Theta (2θ): 6.3°±0.2°, 6.9°±0.2°, 8.4°±0.2°, 9.6°±0.2°, 10.0°±0.2°, 11.0°±0.2°, 12.8°±0.2°, 13.9°±0.2°, 15.2°±0.2°, 15.5°±0.2°, 16.0°±0.2°, 16.6°±0.2°, 17.1°±0.2°, 18.3°±0.2°, 18.7°±0.2°, 19.2°±0.2°, 19.9°±0.2°. The X-ray diffraction patterns are 20.6°±0.2°, 21.0°±0.2°, 21.8°±0.2°, 22.6°±0.2°, 23.2°±0.2°, 23.6°±0.2°, 24.6°±0.2°, 25.7°±0.2°, 27.1°±0.2°, 28.5°±0.2°, 29.7°±0.2°, and 29.8°±0.2°, wherein the X-ray diffraction patterns are obtained using a Cu Kα anode.

[0009] In a more preferred embodiment, the EYP001 tromethamine salt crystal form I results in an X-ray diffraction pattern containing the following peaks:

[0010] The X-ray diffraction pattern was obtained using a Cu Kα anode.

[0011] In another aspect, the EYP001 hydroxylamine salt crystalline form II, which exhibits excellent thermal stability (melting point: 236℃) and is non-hygroscopic, has been isolated.

[0012] Therefore, this document provides a crystalline form (form II) of 4-chloro-5-[4-(2,6-dichlorophenyl)sulfonylpiperazin-1-yl]-1-benzofuran-2-carboxylic acid (EYP001) in an aminobutadiene glycerol salt, wherein the X-ray diffraction pattern of said crystalline form contains peaks at the following diffraction angles 2-Theta (2θ): 9.0°±0.2°, 14.5°±0.2°, 15.9°±0.2°, 17.9°±0.2°, 20.1°±0.2°, 21.4°±0.2°, and 25.2°±0.2°, wherein said X-ray diffraction pattern is obtained using a Cu Kα anode. In a preferred embodiment, the crystalline form II of EYP001 aminobutadiene glycerol salt contains peaks at the following diffraction angles 2-Theta (2θ). Peaks at (2θ): 7.0°±0.2°, 9.0°±0.2°, 11.3°±0.2°, 12.3°±0.2°, 14.0°±0.2°, 14.5°±0.2°, 15.9°±0.2°, 16.3°±0.2°, 16.6°±0.2°, 17.9°±0.2°, 18.7°±0.2°, 20.1°±0.2°, 21.4°±0.2°, 2 2.7°±0.2°, 23.1°±0.2°, 23.8°±0.2°, 24.3°±0.2°, 25.2°±0.2°, 26.4°±0.2°, 26.9°±0.2°, 29.1°±0.2°, 30.4°±0.2°, 31.0°±0.2°, 33.6°±0.2°, 34.9°±0.2°, and 35.8°±0.2°, wherein the X-ray diffraction patterns are obtained using a Cu Kα anode.

[0013] In a more preferred embodiment, the EYP001 tromethamine salt crystal form II results in an X-ray diffraction pattern containing the following peaks:

[0014] The X-ray diffraction pattern was obtained using a Cu Kα anode.

[0015] In another aspect, the monohydrated form of EYP001 aminobutanetriol salt crystalline form III, exhibiting excellent thermal stability (melting point: 237℃), has been isolated.

[0016] Therefore, this document provides a crystalline form (form III) of 4-chloro-5-[4-(2,6-dichlorophenyl)sulfonylpiperazin-1-yl]-1-benzofuran-2-carboxylic acid (EYP001) in an aminobutadiene glycerol salt, wherein the X-ray diffraction pattern of said crystalline form contains peaks at the following diffraction angles 2-Theta (2θ): 11.1°±0.2°, 15.7°±0.2°, 15.9°±0.2°, 17.3°±0.2°, 20.9°±0.2°, 21.2°±0.2°, 21.8°±0.2°, and 27.2°±0.2°, wherein said X-ray diffraction pattern is obtained using a Cu Kα anode. In a preferred embodiment, the crystalline form III of EYP001 aminobutadiene glycerol salt contains peaks at the following diffraction angles 2-Theta (2θ). Peaks at (2θ): 6.3°±0.2°, 7.0°±0.2°, 8.7°±0.2°, 10.5°±0.2°, 11.1°±0.2°, 14.7°±0.2°, 15.9°±0.2°, 17.3°±0.2°, 18.3°±0.2°, 20.0°±0.2°, 20.9°±0.2°, 21.8°±0.2°, 22.7°. The X-ray diffraction patterns are 23.2°±0.2°, 25.1°±0.2°, 25.9°±0.2°, 27.2°±0.2°, 28.3°±0.2°, 29.4°±0.2°, 31.8°±0.2°, 32.4°±0.2°, 35.2°±0.2°, 36.0°±0.2°, and 37.0°±0.2°, wherein the X-ray diffraction patterns are obtained using a Cu Kα anode.

[0017] In a more preferred embodiment, the EYP001 tromethamine salt crystal form III results in an X-ray diffraction pattern containing the following peaks:

[0018] The X-ray diffraction pattern was obtained using a Cu Kα anode.

[0019] Another aspect of the invention is the use of EYP001 tromethamine salt or EYP001 tromethamine salt crystalline form as defined herein, as a medicine or pharmaceutical product.

[0020] Another aspect of the invention is a pharmaceutical or veterinary composition comprising EYP001 tromethamine salt or EYP001 tromethamine salt crystalline form as defined herein, and pharmaceutically acceptable excipients.

[0021] In a particular embodiment, the pharmaceutical or veterinary composition as defined herein further comprises additional therapeutic agents, such as TLR3 agonists, TLR7 agonists, TLR8 agonists, TLR9 agonists, RIG-I modulators, STING agonists, antiviral agents such as bulevirtide, antibacterial agents, interferon or its pegylated form, checkpoint inhibitors such as PD-1 or PD-L1 agonists, ERA, ACE inhibitors, ARBs, RASS antagonists, β-blockers, diuretics, MRA, SGLT2 inhibitors, GLP1 agonists, SGLT1 inhibitors, FGF19, FGF21, DPP-4 inhibitors, PPAR agonists, THR β agonists, FASN, HSD17b13 inhibitors, or combinations thereof.

[0022] Another aspect of the invention is a pharmaceutical composition or veterinary composition as defined herein for treating a disease selected from chronic liver disease, gastrointestinal disease, kidney disease, cardiovascular disease, metabolic disease, infection, cancer, and autoimmune disease. Another aspect of the invention is a method of treating a disease selected from chronic liver disease, gastrointestinal disease, kidney disease, cardiovascular disease, metabolic disease, infection, cancer, and autoimmune disease in a subject with this need, said method comprising administering to the subject an effective amount of a pharmaceutical composition or veterinary composition as defined herein. Another aspect of the invention is the use of a pharmaceutical composition or veterinary composition as defined herein for preparing a medicament for treating a disease selected from chronic liver disease, gastrointestinal disease, kidney disease, cardiovascular disease, metabolic disease, infection, cancer, and autoimmune disease.

[0023] In one particular aspect, the disease is an infection, especially a chronic infection, preferably a viral infection, more preferably an infection caused by hepatitis B virus (HBV), hepatitis C virus (HCV), hepatitis D virus (HDV), herpes simplex virus (HSV), human papillomavirus (HPV) (e.g., condyloma acuminata), varicella-zoster virus, cytomegalovirus (CMV), rhinovirus, hepatitis A virus, hepatitis E virus, Kaposi's sarcoma herpesvirus, coronaviruses (including SARS-CoV1, MERS-CoV, and SARS-CoV2), retroviruses (including HIV), and influenza virus. In particular, pharmaceutical or veterinary compositions for treating the infection are used in combination with TLR3 agonists, TLR7 agonists, TLR8 agonists, TLR9 agonists, RIG-I modulators, STING agonists, antiviral agents such as bufovir dipivoxil, antibacterial agents, interferon or its pegylated form, checkpoint inhibitors such as PD-1 or PD-L1 agonists, or combinations thereof.

[0024] In another specific aspect, the disease is kidney disease, particularly kidney disease including renal fibrosis and / or chronic kidney disease (CKD), such as those selected from hypertension, type 2 diabetes, type 1 diabetes, obesity, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), metabolic (functional disorder)-associated fatty liver disease (MAFLD), aging, infectious glomerulonephritis (especially infections such as syphilis, malaria, hepatitis B, hepatitis C, or HIV), focal segmental glomerulosclerosis, IgA nephropathy, minimal change disease, membranous nephropathy, renal vasculitis, urinary tract obstruction, genetic alterations, autoimmune diseases (such as systemic lupus erythematosus (SLE)), and drug- or toxin-induced kidney disease (such as that caused by drugs such as captopril, NSAIDs, penicillamine, probenecid, butyrazine, anti-T...). NF therapy and thioproline or nephropathy induced by toxins such as inorganic salts (e.g., gold, mercury), AIDS-related nephropathy, ischemic nephropathy, tubulointerstitial nephropathy, hepatorenal syndrome, hydronephrosis, renal dysplasia, medullary cystic nephropathy, medullary sponge kidney, polycystic dysplastic kidney, podocyte disease, renal papillary necrosis, nephritis (including glomerulonephritis, hereditary nephritis, interstitial nephritis, pyelonephritis), nephrocalcinosis, nephrosclerosis, Albert syndrome, cystinosis, classic homocystinuria (HCU), Fabry disease, renal sarcoidosis, diabetic nephropathy, focal segmental glomerulosclerosis (FSGS), hypertensive nephrosclerosis, chronic glomerulonephritis, chronic transplant glomerulonephritis, chronic interstitial nephritis, Sjögren's syndrome, Alagille syndrome, α1-antitrypsin deficiency, and polycystic kidney disease. In a more specific aspect, the disease is chronic kidney disease (CKD). In particular, the pharmaceutical or veterinary composition for treating kidney disease is used in combination with ERA, ACE inhibitors, ARBs, RASS antagonists, β-blockers, diuretics, MRA, SGLT2 inhibitors, GLP1 agonists, or combinations thereof.

[0025] In another specific aspect, the disease is liver disease, especially chronic liver disease, preferably primary biliary cirrhosis or primary biliary cholangitis (PBC), cerebral tendinitis xanthomas (CTX), primary sclerosing cholangitis (PSC), drug-induced cholestasis, intrahepatic cholestasis of pregnancy, parenteral nutrition-associated cholestasis (PNAC), bacterial overgrowth or sepsis-associated cholestasis, autoimmune hepatitis, chronic viral hepatitis, alcoholic liver disease, and nonalcoholic fatty liver disease (NAFLD). Nonalcoholic steatohepatitis (NASH), metabolic-associated fatty liver disease (MAFLD), alcoholic hepatitis, liver transplant-associated graft-versus-host disease, liver regeneration after living donor liver transplantation, congenital liver fibrosis, choledocholithiasis, granulomatous liver disease, intrahepatic or extrahepatic malignancies, Sjögren's syndrome, Alagille syndrome, sarcoidosis, Wilson's disease, Gaucher disease, hemochromatosis, biliary atresia, liver transplant rejection due to bile duct absence, cystic fibrotic liver disease, and α1-antitrypsin deficiency. Specifically, the pharmaceutical or veterinary composition for treating liver diseases is used in combination with SGLT2 inhibitors, GLP1 agonists, SGLT1 inhibitors, FGF19, FGF21, DPP-4 inhibitors, PPAR agonists, THR β agonists, FASN, HSD17b13 inhibitors, or combinations thereof. Attached Figure Description

[0026] Figure 1 XRPD spectra of EYP001 tromethamine salt (form I - sample A), tromethamine counterion, and EYP001 acid form stacked together.

[0027] Figure 2 XRPD plot of EYP001 tromethamine salt (Form I - Sample A) Figure 3 DSC spectrum of EYP001 tromethamine salt (Form I - Sample A).

[0028] Figure 4 TGA spectrum of EYP001 tromethamine salt (Form I - Sample A).

[0029] Figure 5 DVS isothermal adsorption curve of EYP001 tromethamine salt (form I - sample A) at 25℃.

[0030] Figure 6 XRPD spectra of EYP001 tromethamine crystal form I and EYP001 tromethamine crystal form II superimposed.

[0031] Figure 7 XRPD plot of EYP001 tromethamine salt (Form II) Figure 8 DSC spectrum of EYP001 aminobutadiene triol salt (form II).

[0032] Figure 9 TGA spectrum of EYP001 tromethamine salt (form II).

[0033] Figure 10 DVS isothermal adsorption curve of EYP001 aminobutadiene triol salt (form II) at 25℃.

[0034] Figure 11 XRPD spectra of EYP001 tromethamine crystal form I and EYP001 tromethamine crystal form III superimposed.

[0035] Figure 12 XRPD plot of EYP001 tromethamine salt (form III) Figure 13 DSC spectrum of EYP001 aminobutadiene triol salt (form III).

[0036] Figure 14 TGA spectrum of EYP001 aminobutadiene triol salt (form III).

[0037] Figure 15 DVS isothermal adsorption curve of EYP001 aminobutadiene triol salt (form III) at 25℃.

[0038] Figure 16 XRPD spectra of EYP001 aminobutanetriol salts in amorphous and crystalline forms I, II and III. Detailed Implementation

[0039] definition Throughout this specification and the appended claims, unless otherwise expressly stated, the following terms shall have the following meanings.

[0040] As used herein, the terms “about” or “around” will be understood by those skilled in the art and will vary to some extent depending on the context in which they are used. For example, in a particular context, “about” or “around” may refer to a particular term plus or minus up to 10%.

[0041] As used herein, the terms “compound” or “molecule” refer to the EYP001 aminobutanetriol salt according to the present invention, including its forms as disclosed herein.

[0042] As used herein, the terms “treatment,” “treat,” or “treating” refer to any action aimed at improving a patient’s health condition, such as the treatment, prevention, or delay of a disease. In some embodiments, such a term refers to the improvement or eradication of a disease or its accompanying symptoms. In other embodiments, the term refers to minimizing the spread or exacerbation of a disease by administering one or more therapeutic agents to a subject suffering from such a disease.

[0043] As used herein, the terms “subject,” “individual,” or “patient” are used interchangeably and refer to animals, preferably mammals, and even more preferably humans, including adults and children. However, the term “subject” can also refer to non-human animals, particularly mammals such as dogs, cats, horses, cattle, pigs, sheep, and non-human primates.

[0044] The terms “quantity,” “amount,” and “dosage” are used interchangeably in this document and may refer to the absolute quantification of molecules.

[0045] As used herein, the term "therapeutic effect" refers to the effect induced by the active ingredient or pharmaceutical composition according to the invention, which is able to prevent or delay the onset or development of a disease or condition, or to cure a disease or condition, or to reduce the effects of a disease or condition.

[0046] As used herein, the term "effective amount" refers to the amount of an active ingredient or pharmaceutical composition that prevents, removes, or reduces the harmful effects of a disease. The amount to be administered can be adjusted by those skilled in the art based on the subject to be treated, the nature of the disease, etc. In particular, the dosage and regimen may vary depending on the nature, stage, and severity of the disease to be treated, as well as the weight, age, and overall health status of the subject to be treated, and the physician's judgment.

[0047] As used herein, the terms "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" refer to any component present in a pharmaceutical composition other than the active ingredient. Their addition may be intended to impart a specific consistency or other physical or gustatory property to the final product. A pharmaceutically acceptable carrier or pharmaceutically acceptable excipient must not interact with the active ingredient in any way, particularly chemically.

[0048] As used herein, the terms “kit,” “product,” or “combination formulation” specifically refer to “kit of parts” because combination pairs as defined herein can be administered independently or by using different fixed combinations of combination pairs in varying amounts, i.e., simultaneously or at different time points. The parts of the kit can therefore be administered, for example, simultaneously or staggered in timing, i.e., at different time points, and the time intervals between any parts of the kit may be equal or different. The ratio of the total amount of combination pairs to be administered in the combination formulation can be varied. The combination pairs can be administered via the same route or via different routes.

[0049] 4-Chloro-5-[4-(2,6-dichlorophenyl)sulfonylpiperazin-1-yl]-1-benzofuran-2-carboxylic acid (IUPAC name; CAS number 1192171-69-9) is also referred to herein as 4-chloro-5-[4-(2,6-dichlorophenyl-1-sulfonyl)-piperazin-1-yl]-1-benzofuran-2-carboxylic acid or 4-chloro-5-[4-(2,6-dichloro-phenylsulfonyl)-piperazin-1-yl]-benzofuran-2-carboxylic acid, “Vonafexor”, “PLX007”, “EYP001” or “EYP-001”, and has the following formula: .

[0050] The synthesis of the free acid EYP001 was first described by Merck Patent GmbH in Example 136 of WO 2009 / 127321. It is well known that EYP001 can also be prepared by those skilled in the art using other synthetic routes and well-known techniques. EYP001 can even be purchased directly from commercial suppliers.

[0051] The poor solubility of the EYP001 free acid has been a major obstacle to the development of this drug. However, it has now been found that the EYP001 tromethamine salt, as presented in this article, has many improved properties compared to the free acid form and even other salts, including better solubility, and resulting in better bioavailability than the free acid form. Furthermore, it is suitable for pharmaceutical formulations.

[0052] EYP001 tromethamine salt can be prepared according to any method known to those skilled in the art for preparing salts of active ingredients. For example, EYP001 can be dissolved in a solvent, preferably a polar solvent, and an anti-counterion tromethamine can be added. Examples of solvents that can be invoked include, but are not limited to, water, isopropyl acetate, acetonitrile, isopropanol, ethanol, methanol, acetone, tetrahydrofuran, and mixtures thereof. In particular, the solvent is selected from tetrahydrofuran, acetonitrile, water, and ethanol. In a preferred embodiment, the solvent is a mixture of tetrahydrofuran, water, and ethanol. In yet another preferred embodiment, the solvent is tetrahydrofuran. In still another preferred embodiment, the solvent is acetonitrile.

[0053] In one particular embodiment, EYP001 tromethamine salt is prepared by contacting a solution of EYP001 in THF with a solution of tromethamine in water. In yet another particular embodiment of the invention, EYP001 tromethamine salt is prepared by contacting a solution of EYP001 in THF with a solution of tromethamine in ethanol / water.

[0054] A specific embodiment of the present invention is therefore a method for preparing the aminobutadiene triol salt of 4-chloro-5-[4-(2,6-dichlorophenyl)sulfonylpiperazin-1-yl]-1-benzofuran-2-carboxylic acid, comprising the steps of: a) Contacting a solution of 4-chloro-5-[4-(2,6-dichlorophenyl)sulfonylpiperazin-1-yl]-1-benzofuran-2-carboxylic acid in THF with a solution of tromethamine in ethanol / water or in water; and a1) Separate the aminobutane triol salt of 4-chloro-5-[4-(2,6-dichlorophenyl)sulfonylpiperazin-1-yl]-1-benzofuran-2-carboxylic acid obtained in step a).

[0055] Preferably, 4-chloro-5-[4-(2,6-dichlorophenyl)sulfonylpiperazin-1-yl]-1-benzofuran-2-carboxylic acid and tromethamine are used in a stoichiometric ratio of about 1:1.

[0056] Various forms of EYP001 tromethamine salt have been identified. These include various crystalline and amorphous forms.

[0057] The present invention also provides crystalline forms of EYP001 tromethamine salt (forms I-IV). These crystalline forms also exhibit physicochemical properties favorable for pharmaceutical dosage forms. In particular, crystalline forms I, II, and III have excellent thermal stability (above 180°C), and crystalline forms I and II are non-hygroscopic.

[0058] The crystalline forms described herein can be identified and / or characterized by a variety of analytical techniques known to those skilled in the art. Such techniques include, but are not limited to, X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), dynamic vapor adsorption (DVS), and / or IR spectroscopy.

[0059] In particular, the crystalline form of EYP001 aminobutadiene trioxide can be characterized by X-ray diffraction patterns.

[0060] Specifically, the X-ray diffraction pattern of one crystalline form (Form I) contains peaks at the following diffraction angles 2-Theta (2θ): 15.2°±0.2°, 16.0°±0.2°, 17.1°±0.2°, 18.3°±0.2°, 18.7°±0.2°, 20.6°±0.2°, 22.6°±0.2°, and 23.2°±0.2°, wherein the X-ray diffraction pattern is obtained using a Cu Kα anode. More specifically, the X-ray diffraction pattern of the crystalline form contains peaks at the following diffraction angles 2-Theta (2θ): 6.3°±0.2°, 6.9°±0.2°, 8.4°±0.2°, 9.6°±0.2°, 10.0°±0.2°, 11.0°±0.2°, 12.8°±0.2°, 13.9°±0.2°, 15.2°±0.2°, 15.5°±0.2°, 16.0°±0.2°, 16.6°±0.2°, 17.1°±0.2°, 18.3°±0.2°, 18.7°±0.2°, 19.2°±0.2°, 19.9°±0.2°. The X-ray diffraction patterns are 20.6°±0.2°, 21.0°±0.2°, 21.8°±0.2°, 22.6°±0.2°, 23.2°±0.2°, 23.6°±0.2°, 24.6°±0.2°, 25.7°±0.2°, 27.1°±0.2°, 28.5°±0.2°, 29.7°±0.2°, and 29.8°±0.2°, wherein the X-ray diffraction patterns are obtained using a Cu Kα anode. More specifically, the X-ray diffraction patterns of this crystalline form obtained using a Cu Kα anode contain the peaks listed in Table 1 below or as shown below. Figure 2 The peaks shown in the X-ray diffraction pattern are as follows.

[0061] Table 1: X-ray peaks of EYP001 tromethamine salt crystal form I

[0062] Specifically, the X-ray diffraction pattern of one crystalline form (form II) contains peaks at the following diffraction angles 2-Theta (2θ): 9.0°±0.2°, 14.5°±0.2°, 15.9°±0.2°, 17.9°±0.2°, 20.1°±0.2°, 21.4°±0.2°, and 25.2°±0.2°, wherein the X-ray diffraction pattern was obtained using a Cu Kα anode. More specifically, the X-ray diffraction pattern of the crystalline form contains peaks at the following diffraction angles 2-Theta (2θ). Peaks at (2θ): 7.0°±0.2°, 9.0°±0.2°, 11.3°±0.2°, 12.3°±0.2°, 14.0°±0.2°, 14.5°±0.2°, 15.9°±0.2°, 16.3°±0.2°, 16.6°±0.2°, 17.9°±0.2°, 18.7°±0.2°, 20.1°±0.2°, 21.4°±0.2°, 2 2.7°±0.2°, 23.1°±0.2°, 23.8°±0.2°, 24.3°±0.2°, 25.2°±0.2°, 26.4°±0.2°, 26.9°±0.2°, 29.1°±0.2°, 30.4°±0.2°, 31.0°±0.2°, 33.6°±0.2°, 34.9°±0.2°, and 35.8°±0.2°, wherein the X-ray diffraction patterns are obtained using a Cu Kα anode. More particularly, the X-ray diffraction patterns of this crystalline form obtained using a Cu Kα anode contain the peaks listed in Table 2 below or as shown below. Figure 7 The peaks shown in the X-ray diffraction pattern are as follows.

[0063] Table 2: X-ray peaks of EYP001 tromethamine salt crystal form II

[0064] Specifically, the X-ray diffraction pattern of one crystalline form (form III) contains peaks at the following diffraction angles 2-Theta (2θ): 11.1°±0.2°, 15.7°±0.2°, 15.9°±0.2°, 17.3°±0.2°, 20.9°±0.2°, 21.2°±0.2°, 21.8°±0.2°, and 27.2°±0.2°, wherein the X-ray diffraction pattern was obtained using a Cu Kα anode. More specifically, the X-ray diffraction pattern of the crystalline form contains peaks at the following diffraction angles 2-Theta (2θ). Peaks at (2θ): 6.3°±0.2°, 7.0°±0.2°, 8.7°±0.2°, 10.5°±0.2°, 11.1°±0.2°, 14.7°±0.2°, 15.9°±0.2°, 17.3°±0.2°, 18.3°±0.2°, 20.0°±0.2°, 20.9°±0.2°, 21.8°±0.2°, 22.7°. The X-ray diffraction patterns are 23.2°±0.2°, 25.1°±0.2°, 25.9°±0.2°, 27.2°±0.2°, 28.3°±0.2°, 29.4°±0.2°, 31.8°±0.2°, 32.4°±0.2°, 35.2°±0.2°, 36.0°±0.2°, and 37.0°±0.2°, wherein the X-ray diffraction patterns are obtained using a Cu Kα anode. More specifically, the X-ray diffraction patterns of this crystalline form obtained using a Cu Kα anode contain the peaks listed in Table 3 below or as shown below. Figure 12 The peaks shown in the X-ray diffraction pattern are as follows.

[0065] Table 3: X-ray peaks of EYP001 tromethamine salt crystal form III

[0066] Those skilled in the art will understand that the measurement error of the obtainable X-ray diffraction pattern depends on the measurement conditions employed. In particular, it is well known that the intensity in an X-ray diffraction pattern can fluctuate with the measurement conditions employed, the shape or morphology of the particles, and the crystal size distribution. It should also be understood that relative intensity may also vary with experimental conditions, and therefore the exact order of intensity should not be considered. Furthermore, the measurement error of the diffraction angle in conventional X-ray diffraction patterns is typically about ±0.02° (2θ) or less, preferably about ±0.01°. Therefore, it should be understood that the crystal form of the present invention is not limited to providing values ​​respectively relative to… Figure 2 , 7 Crystal forms of X-ray diffraction patterns that are exactly the same as those shown in Figure 12 or described in Tables 1, 2, and 3. For example, providing X-ray diffraction patterns that are identical to those shown in Figure 12 or described in Tables 1, 2, and 3. Figure 2 Any crystal form with substantially the same X-ray diffraction pattern disclosed in or described in Table 1 falls within the scope of this invention. This also applies to crystals providing X-ray diffraction patterns similar to those disclosed in Table 1. Figure 7 and 12 Any crystal form with substantially the same X-ray diffraction pattern disclosed in or described in Tables 2 and 3. The ability to determine the substantial identity of the X-ray diffraction patterns is within the capabilities of a person skilled in the art.

[0067] The crystal form of EYP001 aminobutadiene triol salt can also be characterized by differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), and dynamic vapor adsorption (DVS).

[0068] In one particular embodiment, the crystalline form of EYP001 tromethamine (Form I) has a melting point above 180°C, as determined by differential scanning calorimetry (DSC). In another particular embodiment, the crystalline form of EYP001 tromethamine (Form I) shows no significant mass loss as determined by thermogravimetric analysis (TGA) in the range of 20°C to 182°C. In another particular embodiment, the crystalline form of EYP001 tromethamine (Form I) is non-hygroscopic. More specifically, dynamic vapor adsorption (DVS) analysis of this crystalline form I of the present invention shows a water absorption rate of only 1.3% at 25°C / 60% RH.

[0069] In yet another specific embodiment, the EYP001 tromethamine crystalline form (form II) has a melting point of approximately 236°C according to differential scanning calorimetry (DSC). In one specific embodiment, the EYP001 tromethamine crystalline form (form II) showed no significant mass loss as determined by thermogravimetric analysis (TGA) at an initial temperature of 220°C. In one specific embodiment, the EYP001 tromethamine crystalline form (form II) is non-hygroscopic. More particularly, dynamic vapor adsorption (DVS) analysis of this crystalline form II of the present invention showed a water absorption rate of only 0.3% at 25°C / 60% RH.

[0070] In yet another specific embodiment, the EYP001 tromethamine crystalline form (Form III) has a melting point of approximately 237°C according to differential scanning calorimetry (DSC). In one specific embodiment, the EYP001 tromethamine crystalline form (Form III) is in a hydrated state. More specifically, dynamic vapor adsorption (DVS) analysis of this crystalline form III of the present invention shows that the bulk can be considered to exist in a hydrated state.

[0071] One or more crystalline forms of the present invention can be prepared by a variety of methods, including, for example, crystallization or recrystallization from a suitable solvent, sublimation, growth from a melt, solid-state transformation from another phase, crystallization from a supercritical fluid, and spray drying. Techniques for crystallizing or recrystallizing crystalline forms from solvent mixtures include, for example, evaporating the solvent, lowering the temperature of the solvent mixture, seeding the crystal with a supersaturated solvent mixture of molecules and / or salts, freeze-drying the solvent mixture, and adding an antisolvent (anti-solvent) to the solvent mixture. Crystals (including polymorphs), preparation methods, and characterization of drug crystals are discussed in Solid-State Chemistry of Drugs, SR Byrn, RR Pfeiffer, and JG Stowell, 2nd edition, SSCI, West Lafayette, Ind. (1999). For crystallization techniques using solvents, the choice of one or more solvents typically depends on one or more factors, such as the solubility of the compound, the crystallization technique, the vapor pressure of the solvent, the viscosity of the solvent, and the toxicity of the solvent. Combinations of solvents can be used; for example, the compound can be dissolved in a first solvent to provide a solution, followed by the addition of an antisolvent to reduce the solubility of the compound in the solution and to provide crystal formation. An antisolvent is a solvent in which a compound has low solubility. In one particular embodiment, the solvent is selected from water, isopropyl acetate, ethyl acetate, dimethyl sulfoxide, heptane, acetonitrile, isopropanol, methanol, ethanol, acetone, tetrahydrofuran, and mixtures thereof. In a preferred embodiment, the solvent is selected from water, tetrahydrofuran, acetonitrile, and mixtures thereof. In yet another preferred embodiment, the antisolvent is selected from ethanol and acetone.

[0072] One aspect of the present invention is a method for preparing the crystalline form of 4-chloro-5-[4-(2,6-dichlorophenyl)sulfonylpiperazin-1-yl]-1-benzofuran-2-carboxylic acid in the form of anhydrous triol salt, comprising the steps of: a) Contact a solution of 4-chloro-5-[4-(2,6-dichlorophenyl)sulfonylpiperazin-1-yl]-1-benzofuran-2-carboxylic acid in THF with a solution of tromethamine in ethanol / water or in water; b) Optionally triggering the formation of the crystalline form, and / or optional improving the crystallinity of the crystalline form; and c) Separate the crystalline form obtained in step b) or a).

[0073] A particular aspect of the present invention is a method for preparing the crystalline form of the aminobutadiene triol salt of 4-chloro-5-[4-(2,6-dichlorophenyl)sulfonylpiperazin-1-yl]-1-benzofuran-2-carboxylic acid, comprising the steps of: a) Contact a solution of 4-chloro-5-[4-(2,6-dichlorophenyl)sulfonylpiperazin-1-yl]-1-benzofuran-2-carboxylic acid in THF with a solution of tromethamine in water; b) Optionally triggering the formation of the crystalline form, and / or optionally improving the crystallinity of the crystalline form, preferably by adding ethanol; and c) Separate the crystalline form obtained in step b) or a).

[0074] In one particular embodiment, step a) is carried out in solution, specifically by suspending EYP001 in THF and suspending tromethamine in ethanol / water, preferably water. In yet another particular embodiment, the stoichiometry of 4-chloro-5-[4-(2,6-dichlorophenyl)sulfonylpiperazin-1-yl]-1-benzofuran-2-carboxylic acid:tromethamine in step a) is about 1:1.

[0075] In one particular embodiment, when step a) is carried out in solution, step b) may be carried out, for example, by adding an antisolvent such as ethanol. Step b) may also be carried out by seeding the solution with crystals of the desired crystalline form and / or applying one or more temperature cycles. In one embodiment, when step a) is carried out in solution, step b) is carried out by adding an antisolvent such as ethanol, or by seeding the solution with crystals of the desired crystalline form and then applying one or more temperature cycles. The latter step is designed to improve crystallinity and therefore the filterability and chemical purity of the final product.

[0076] In one particular embodiment, step c) may be performed, for example, by evaporating the reaction solvent, by filtration, or by centrifugation. In yet another particular embodiment, step c) may also include one or more washing steps. Each step, each part of a step (e.g., the first or second part of step b), and / or each combination of steps (e.g., a combination of step b and step c) of the method of the present invention may be performed once or may be repeated several times in the method of the present invention, independent of other steps or part of a step.

[0077] For example, in step b), the temperature cycle may be performed once, or it may be repeated several times, preferably between two and five times, more preferably two or three times. Repeating step b) continuously allows for particularly increased crystallinity of the form.

[0078] The crystalline form obtained by the method described is another aspect of the present invention. This can be crystalline form I.

[0079] EYP001 aminobutadiene triol salt crystal form II can be prepared starting from crystal form I.

[0080] Another specific aspect of the present invention is a method for preparing EYP001 hydroxylamine salt crystalline form II, the method comprising the steps of: a) Stir EYP001 hydroxybutyrate crystal form I in THF at a temperature between 40 and 60°C, preferably about 50°C, for several hours, such as between 4 and 18 hours, 4 and 16 hours, 6 and 14 hours, preferably between 8 and 12 hours. b) Separate the solid obtained in step a), wash and dry it; and c) The EYP001 aminobutadiene triol salt crystal form II was isolated.

[0081] EYP001 tromethamine salt crystalline form III can be prepared from an amorphous form, which can be obtained by dissolving EYP001 tromethamine salt crystalline form I in a mixture of THF / water and freeze-drying the mixture.

[0082] Another specific aspect of the present invention is a method for preparing EYP001 hydroxylamine trioxide crystal form III, the method comprising the steps of: a) Add acetonitrile to the amorphous form of EYP001 tromethamine salt to obtain a suspension, and allow it to stand at 4°C for several hours, such as between 1 and 6 hours or 2 and 5 hours. b) Separate the solid obtained in step a) by filtration, wash and dry; and c) The EYP001 aminobutadiene triol salt crystal form III was isolated.

[0083] Another specific aspect of the present invention is a method for preparing the amorphous form of EYP001 tromethamine salt, the method comprising the steps of: a) Dissolve EYP001 aminobutadiene trioxide crystal form I in a mixture of THF / water; b) Freeze-dry the mixture; and c) Separate the amorphous form.

[0084] This document also provides pharmaceutical or veterinary compositions comprising EYP001 tromethamine salt. These compositions optionally comprise at least one pharmaceutically acceptable carrier or excipient. The EYP001 tromethamine salt may be in crystalline form, for example, in form I, II, or III.

[0085] The compounds, pharmaceutical compositions, or veterinary compositions defined herein may be administered via any conventional route of administration. In particular, the compounds, pharmaceutical compositions, or veterinary compositions may be administered topically, enterically, orally, parenterally, intranasally, intravenously, intra-arterially, intramuscularly, subcutaneously, or intraocularly.

[0086] Preferably, the compound according to the invention, or the pharmaceutical composition or veterinary composition according to the invention, is administered via enteral or parenteral route. When administered parenterally, the compound, pharmaceutical composition, or veterinary composition is preferably administered via intravenous route. When administered enterally, the compound, pharmaceutical composition, or veterinary composition is preferably administered via oral route.

[0087] Pharmaceutical compositions comprising the compounds of the present invention are formulated in accordance with standard pharmaceutical practices known to those skilled in the art (Lippincott Williams & Wilkins, 2000 and Encyclopedia of Pharmaceutical Technology, edited by J. Swarbrick and JC Boylan, 1988–1999, Marcel Dekker, New York).

[0088] For oral administration, the composition can be formulated into conventional oral dosage forms such as tablets, capsules, powders, granules, and liquid formulations such as syrups, elixirs, and concentrated drops. Non-toxic solid carriers or diluents can be used, including, for example, pharmaceutical-grade mannitol, lactose, starch, magnesium stearate, sodium saccharin, talc, cellulose, glucose, sucrose, magnesium carbonate, etc. For compressed tablets, a binder is also required; the binder is an agent that imparts cohesiveness to the powdered material. For example, starch, gelatin, sugars (such as lactose or dextrose), and natural or synthetic gums can be used as binders. Disintegrants are also required in tablets to promote tablet disintegration. Disintegrants include starch, clay, cellulose, alginate, gums, and cross-linked polymers. In addition, tablets include lubricants and flow aids to prevent tablet material from adhering to surfaces during preparation and to improve the flow properties of the powdered material during preparation. Colloidal silica is most commonly used as a flow aid, and compounds such as talc or stearic acid are most commonly used as lubricants.

[0089] For transdermal application, the composition can be formulated as an ointment, cream, or gel and can be enhanced with appropriate penetrants or detergents, such as dimethyl sulfoxide, dimethyl acetamide, and dimethylformamide.

[0090] For transmucosal application, nasal sprays, rectal suppositories, or vaginal suppositories can be used. The active compound can be incorporated into any known suppository base using methods known in the art. Examples of such bases include cocoa butter, polyethylene glycol (carbomer wax), polyethylene sorbitan monostearate, and mixtures of these with other compatible materials used to modify melting point or dissolution.

[0091] The pharmaceutical or veterinary compositions according to the invention can be formulated to release the active drug substantially immediately after administration or at any predetermined time or period after administration.

[0092] The compounds, pharmaceutical compositions, or veterinary compositions according to the invention can be administered as a single dose or in multiple doses. Preferably, treatment is administered periodically, preferably daily to monthly, more preferably daily to bi-weekly, more preferably daily to weekly, and even more preferably daily. In one particular embodiment, treatment is administered daily, optionally once, twice, or three times a day. In one particular aspect, treatment is administered at least twice a day, particularly twice or three times a day. In an alternative aspect, treatment is administered once a day. The duration of treatment using the compounds or pharmaceutical compositions according to the invention can be several weeks, months, or even years. In particular, the duration of treatment can continue until the disease subsides.

[0093] The amount of the compound, pharmaceutical composition, or veterinary composition of the present invention to be administered can be determined by standard procedures well known to those skilled in the art. To determine an appropriate dosage, the patient's physiological data (e.g., age, body size, and weight) and the route of administration must be considered to administer a therapeutically effective amount to the patient. In one particular aspect, the total compound dose per administration of the compound, pharmaceutical composition, or veterinary composition according to the present invention is between 0.00001 and 1 g. Optionally, the daily dose of tromethamine salt, for example, in crystalline form as defined herein, can vary within a wide range: 10 to 1,000 mg / adult / day, 25 to 1,000 mg / adult / day, 50 to 1,000 mg / adult / day, 25 to 800 mg / adult / day, 50 to 800 mg / adult / day, 50 to 600 mg / adult / day, or 100 to 600 mg / adult / day. Preferably, the daily dose of tromethamine salt in crystalline form as defined herein is 1 to 500 mg / day, 2 to 450 mg / day, 5 to 450 mg / day, 10 to 450 mg / day, 20 to 450 mg / day, 25 to 450 mg / day, 50 to 450 mg / day, 100 to 450 mg / day, 150 to 450 mg / day, 25 to 400 mg / day, 50 to 400 mg / day, 100 to 400 mg / day, 150 to 400 mg / day, 25 to 350 mg / day, 50 to 350 mg / day, 100 to 350 mg / day, 150 to 350 mg / day, 200 to 450 mg / day, 200 to 350 mg / day, 250 to 450 mg / day, 250 to 40 ... mg / day or in the range of 250 to 350 mg / day. Optionally, the composition, dosage unit or dosage form comprises 2, 5, 10, 15, 25, 50, 75, 100, 200, 300, 400 and 500 mg of tromethamine salt, for example in a crystalline form as defined herein, in order to adjust the dose according to the symptoms of the patient to be treated. The formulation typically contains about 5 mg to about 800 mg of tromethamine (e.g., in crystalline form as defined herein), about 25 mg to about 800 mg of tromethamine (e.g., in crystalline form as defined herein), about 25 mg to about 500 mg of tromethamine (e.g., in crystalline form as defined herein), about 10 mg to about 500 mg of tromethamine (e.g., in crystalline form as defined herein), about 25 mg to about 450 mg of tromethamine (e.g., in crystalline form as defined herein), about 50 mg to about 400 mg of tromethamine (e.g., in crystalline form as defined herein), or about 50 mg to about 300 mg of tromethamine (e.g., in crystalline form as defined herein).The crystal form can be form I, form II or form III.

[0094] The form, route of administration, and dosage of the pharmaceutical or veterinary composition according to the present invention can be adjusted by those skilled in the art based on the type and severity of the disease and on the patient (particularly their age, weight, sex, and general physical condition).

[0095] Another aspect of the invention relates to a tromethamine salt, for example in a crystalline form as defined herein, used as a pharmaceutical or medicament, or a pharmaceutical or veterinary composition, optionally comprising a pharmaceutically acceptable carrier or excipient. The invention also relates to the use of a tromethamine salt (e.g., in a crystalline form as defined herein) as a pharmaceutical or medicament. The invention further relates to a method of treating a disease in a subject, wherein a therapeutically effective amount of a tromethamine salt (e.g., in a crystalline form as defined herein) is administered to the subject in need. The invention also relates to the use of a tromethamine salt (e.g., in a crystalline form as defined herein) in the preparation of a pharmaceutical product. The invention also relates to a pharmaceutical or veterinary composition comprising a tromethamine salt (e.g., in a crystalline form as defined herein), used as a pharmaceutical product. The crystalline form may be form I, form II, or form III.

[0096] Optionally, tromethamine salts (e.g., in a crystalline form as defined herein) or pharmaceutical or veterinary compositions comprising them are used in combination with additional therapeutic agents for the treatment of a disease. Optionally, pharmaceutical or veterinary compositions comprising tromethamine salts (e.g., in a crystalline form as defined herein) also comprise additional therapeutic agents. Optionally, the present invention relates to a product or kit comprising tromethamine salts (e.g., in a crystalline form as defined herein) and additional therapeutic agents as a combination formulation for simultaneous, separate, or sequential use, particularly for the treatment of a disease. Optionally, the present invention relates to a combination formulation comprising tromethamine salts (e.g., in a crystalline form as defined herein) and additional therapeutic agents for simultaneous, separate, or sequential use, particularly for the treatment of a disease. Optionally, the present invention relates to the use of tromethamine salts (e.g., in a crystalline form as defined herein) or pharmaceutical or veterinary compositions comprising them in the preparation of a medicine for treating a disease in combination with additional therapeutic agents. Optionally, the present invention relates to a method of treating a disease in a subject, wherein a therapeutically effective amount of a tromethamine salt (e.g., in a crystalline form as defined herein) and a therapeutically effective amount of another therapeutic agent are administered to the subject in need. The other therapeutic agent used in combination with the tromethamine salt (e.g., in a crystalline form as defined herein) may be any of the other therapeutic agents disclosed below, regardless of the disease to be treated. The crystalline form may be form I, form II, or form III.

[0097] Optionally, the additional therapeutic agents may be selected from TLR3 agonists, TLR7 agonists, TLR8 agonists, TLR9 agonists, RIG-I modulators, STING agonists, antiviral agents such as bubevirtide, antibacterial agents, interferon or its pegylated form, checkpoint inhibitors such as PD-1 or PD-L1 agonists, ERA, ACE inhibitors, ARBs, RASS antagonists, β-blockers, diuretics, MRA, SGLT2 inhibitors, GLP1 agonists, SGLT1 inhibitors, FGF19, FGF21, DPP-4 inhibitors, PPAR agonists, THR β agonists, FASN, HSD17b13 inhibitors, or combinations thereof.

[0098] The diseases to be treated can be selected from chronic liver disease, gastrointestinal disease, kidney disease, cardiovascular disease, metabolic disease, infection, cancer and autoimmune disease.

[0099] In the first aspect, the disease is an infection, especially a chronic infection. The infection can be a viral infection, a bacterial infection (especially a mycobacterial infection), or a protozoan infection. In one particular aspect, the infection is a viral infection.

[0100] The additional therapeutic agent may be any medicine that can be used to treat infection, such as TLR3 agonists, TLR7 agonists, TLR8 agonists, TLR9 agonists, RIG-I modulators, STING agonists, antiviral agents such as bufovir dipivoxil, antibacterial agents, interferon or its pegylated form, checkpoint inhibitors such as PD-1 or PD-L1 agonists or combinations thereof. In a very specific aspect, the additional therapeutic agents are TLR3 agonists, such as polyI:C (polyribonucleic acid: polycytidine), polyA:U (poly(adenosine monophosphate-uridine monophosphate), polyICLC (polyinosine monophosphate-polycytidine monophosphate-poly-L-lysine carboxymethyl cellulose complex or Hiltonol), polyI:polyC12U (polyIC12U, Ampligen or Rintatolimod), Riboxxol (RGIC®50), RIBOXXIM (RGIC®100), APOXXIM, TL-532, ARNAX, IPH3102, MCT-465 and MCT-485.

[0101] The non-exhaustive list of viral infections includes infections caused by hepatitis B virus (HBV), hepatitis C virus (HCV), hepatitis D virus (HDV), herpes simplex virus (HSV), human papillomavirus (HPV) (e.g., genital warts), varicella-zoster virus, cytomegalovirus (CMV), rhinovirus, hepatitis A virus, hepatitis E virus, Kaposi's sarcoma herpesvirus, coronaviruses (including SARS-CoV1, MERS-CoV, and SARS-CoV2), retroviruses (including HIV), and influenza virus.

[0102] In one specific aspect, viral infection is an infection caused by hepatitis B virus (HBV), particularly chronic HBV infection or chronic HBV hepatitis. Optionally, additional therapeutic agents to be used in combination to treat HBV are selected from polymerase inhibitors such as L-nucleosides, deoxyguanosine analogs and nucleoside phosphonates, nucleoside analogs such as lamivudine (Epivir), adefovir (Hepsera), tenofovir (Viread), telbivudine (Tyzeka), entecavir (Baraclude), emtricitabine, and interferons such as interferon alpha-2a and pegylated interferon alpha-2a (Pegasys) and interferon alpha-2b (Viraferon Peg or Introna).

[0103] In one specific sense, viral infection refers to infection caused by hepatitis D virus (HDV), particularly chronic HDV infection or chronic HDV hepatitis.

[0104] Optionally, additional therapeutic agents to be used in combination to treat HDV infection or co-infection of HBV and HDV are selected from the following: - Interferons, such as interferon α (IFN-α), interferon λ or their PEGylated forms, preferably selected from IFN-α (such as IFN-α1a, IFN-α1b, IFN-α2a, IFN-α2b) and IFN-λ (such as IFN-λ1a or their PEGylated forms), more preferably PEG-IFN-α2a (e.g., Pegasys), PEG-IFN-α2b (e.g., ViraferonPeg or Introna) or PEG-IFN-λ1a; - Antiviral drugs specifically targeting HDV, such as nucleoside analogs, isoprenoidation inhibitors or farnesyltransferase inhibitors, preferably ribavirin, ritonavir, lonafanib and EBP 921, more preferably ritonavir, lonafanib or combinations thereof; - HBV-specific antiviral agents, such as nucleoside analogs like lamivudine, adefovir, telbivudine, entecavir, tenofovir, and emtricitabine; - Antiviral agents specifically targeting HBV and HDV, such as nucleoside analogs, nucleic acid polymers, HBsAg secretion inhibitors or NTCP inhibitors, more preferably buleviride (myrcludex B or Hepcludex), ezetimibe, nucleic acid polymer REP2139 and nucleic acid polymer REP 2165; - Viral expression inhibitors that target HBV transcripts, especially siRNAs or antisense oligonucleotides, such as JNJ-3989 (ARO-HBV), VIR-2218, RG6346 (DCR-HBVS, RO7445482), Bepirovirsen (GSK3228836, ISIS 505358) or RO7062931; - Antibodies against HBV, especially those against HBsAg, such as VIR-3434; and - Combinations such as PEG-IFN-λ and lonafab, VIR-3434 and VIR-2218, ritonavir and lonafab (optionally with PEG-IFN-α2a), JNJ-3989 (ARO-HBV) and entecavir, JNJ-3989 (ARO-HBV) and tenofovir, VIR-2218 and PEG-IFN-α, or buleviride and lonafab.

[0105] In the second aspect, the disease is kidney disease. In one specific aspect, kidney disease includes renal fibrosis, including glomerular sclerosis and / or tubulointerstitial fibrosis.

[0106] In one aspect, the subject to be treated has renal fibrosis. Renal fibrosis can be diagnosed based on a renal biopsy. Alternatively, it can be diagnosed based on alternative analyses such as magnetic resonance imaging (MRI) or urinary proteomics (e.g., CKD273).

[0107] In one specific aspect, the kidney disease is chronic kidney disease. Preferably, the subject has renal fibrosis and suffers from chronic kidney disease.

[0108] Chronic kidney disease (CKD) is defined as the presence of kidney damage (usually measured as urinary albumin excretion ≥30 mg / day or equivalent) or decreased kidney function (defined as estimated glomerular filtration rate [eGFR] <60 mL / min / 1.73 m2) lasting for three months or more, regardless of the cause.

[0109] In one particular aspect, CKD is CKD staged based on glomerular filtration rate (eGFR) as shown in Table A, selected from G1, G2, G3a, G3b, G4, or G5, preferably G1, G2, G3a, G3b, or G4, more preferably G2, G3a, G3b, or G4, and even more particularly G2, G3a, or G3b. Preferably, the subject has renal fibrosis.

[0110] In another specific aspect, CKD is CKD staging based on proteinuria (ACR), selected from A1, A2 or A3.

[0111] Table A: Stages of CKD

[0112] Optionally, CKD has a selection from 1 as defined in Table A. The CKD may be staged in one, two, or three phases as defined in Table A. Alternatively, the CKD may be staged in one or two phases as defined in Table A.

[0113] Optionally, the subject had renal fibrosis or CKD and the disease was selected from hypertension, type 2 diabetes, type 1 diabetes, obesity, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), metabolic (functional)-associated fatty liver disease (MAFLD), aging, infectious glomerulonephritis (especially infections such as syphilis, malaria, hepatitis B, hepatitis C, or HIV), focal segmental glomerulosclerosis, IgA nephropathy, minimal change disease, membranous nephropathy, renal vasculitis, urinary tract obstruction, genetic alterations, autoimmune diseases (such as systemic lupus erythematosus (SLE)), and drug- or toxin-induced nephropathy (such as that caused by drugs such as captopril, NSAIDs, penicillamine, probenecid, butyrazine, anti-TNF therapy, and thioproline, or by toxins). Examples of kidney diseases include: inorganic salt-induced nephropathy (e.g., gold, mercury), AIDS-related nephropathy, ischemic nephropathy, tubulointerstitial nephropathy, hepatorenal syndrome, hydronephrosis, renal dysplasia, medullary cystic nephropathy, medullary sponge kidney, polycystic dysplastic kidney, podocyte disease, renal papillary necrosis, nephritis (including glomerulonephritis, hereditary nephritis, interstitial nephritis, pyelonephritis), nephrocalcinosis, nephrosclerosis, Albert syndrome, cystinosis, classic homocystinuria (HCU), Fabry disease, renal sarcoidosis, diabetic nephropathy, focal segmental glomerulosclerosis (FSGS), hypertensive nephrosclerosis, chronic glomerulonephritis, chronic transplant glomerulonephritis, chronic interstitial nephritis, Sjögren's syndrome, Alagille syndrome, α1-antitrypsin deficiency, and polycystic kidney disease.

[0114] In one specific aspect, the subject had renal fibrosis or CKD, and the disease was selected from hypertension, type 2 diabetes, type 1 diabetes, obesity, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), metabolic (functional)-associated fatty liver disease (MAFLD), aging, infectious glomerulonephritis (especially infections such as syphilis, malaria, hepatitis B, hepatitis C, or HIV), focal segmental glomerulosclerosis, IgA nephropathy, minimal change disease, membranous nephropathy, renal vasculitis, urinary tract infection, urinary tract obstruction, genetic alterations, autoimmune diseases (such as systemic lupus erythematosus (SLE)), and drug- or toxin-induced nephropathy (such as nephropathy induced by drugs such as captopril, NSAIDs, penicillamine, probenecid, butyrazine, anti-TNF therapy, and thioproline, or nephropathy induced by toxins such as inorganic salts (e.g., gold, mercury). Optionally, the subject had renal tubulointerstitial fibrosis.

[0115] In another specific aspect, the subject has renal fibrosis or CKD and the disease is a systemic disease affecting the kidneys, such as those selected from: hypertension, type 2 diabetes, type 1 diabetes, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), metabolic (functional)-associated fatty liver disease (MAFLD), infectious glomerulonephritis (especially infections such as syphilis, malaria, hepatitis B, hepatitis C, or HIV), renal vasculitis, autoimmune diseases (such as systemic lupus erythematosus (SLE)), and drug- or toxin-induced nephropathy (such as neuropathy induced by drugs such as captopril, NSAIDs, penicillamine, probenecid, butyrazine, anti-TNF therapy, and thioproline, or by toxins such as inorganic salts (e.g., gold, mercury). Optionally, the subject has renal tubulointerstitial fibrosis.

[0116] Optionally, the subject has type 2 diabetes or type 1 diabetes. The subject may have CKD as defined in Table A above, stage 1, 2, or 3. Optionally, the subject has renal fibrosis, especially tubulointerstitial fibrosis.

[0117] Optionally, the subject has systemic lupus erythematosus (SLE). The subject may have stage 1, 2, or 3 CKD as defined in Table A above. In this context, the subject may have class III, IV, V, or VI lupus nephropathy. Optionally, the subject has renal fibrosis, especially tubulointerstitial fibrosis.

[0118] Optionally, the subject has NASH or NAFLD. The subject may have CKD as defined in Table A above, stage 1, 2, or 3. Nonalcoholic steatohepatitis (NASH) is a disease characterized by excessive fat accumulation, inflammation, and ballooning degeneration of hepatocytes, with or without fibrosis in the liver. Additionally, some subjects affected by NASH may also have chronic kidney disease. The compounds of the present invention may be particularly significant for these specific subjects. Indeed, the compounds of the present invention can significantly reduce inflammation and fibrosis in the liver in a dose-dependent manner, and they can also significantly inhibit fibrosis in the kidneys, even reversing existing fibrosis, and also reducing kidney inflammation. Therefore, the compounds of the present invention can be used to protect subjects with hepatic NASH and renal lesions or to treat subjects with NASH to limit, slow, or reverse liver and kidney lesions. Furthermore, the subject is particularly a subject with NASH or NAFLD and renal fibrosis. Optionally, the subject is a subject with both NASH or NAFLD and CKD. Optionally, the subjects had renal fibrosis, especially tubulointerstitial fibrosis.

[0119] Optionally, the kidney disease is selected from AIDS-related kidney disease, ischemic nephropathy, tubulointerstitial nephropathy, hepatorenal syndrome, hydronephrosis, renal dysplasia, medullary cystic nephropathy, medullary sponge kidney, polycystic dysplastic kidney, podocyte disease, renal papillary necrosis, nephritis (including glomerulonephritis, hereditary nephritis, interstitial nephritis, pyelonephritis), nephrocalcinosis, nephrosclerosis, Albert syndrome, cystinosis, classic homocystinuria (HCU), Fabry disease, renal sarcoidosis, diabetic nephropathy, focal segmental glomerulosclerosis (FSGS), hypertensive nephrosclerosis, chronic glomerulonephritis, chronic transplant glomerulonephritis, chronic interstitial nephritis, Sjögren's syndrome, Alagille syndrome, α1-antitrypsin deficiency, and polycystic kidney disease.

[0120] Optionally, kidney disease is selected from AIDS-related kidney disease, ischemic nephropathy, tubulointerstitial nephropathy, hepatorenal syndrome, hydronephrosis, renal dysplasia, medullary cystic nephropathy, medullary sponge kidney, polycystic dysplastic kidney, podocyte disease, renal papillary necrosis, nephritis (including glomerulonephritis, hereditary nephritis, interstitial nephritis, pyelonephritis), nephrocalcinosis, nephrosclerosis, Allport syndrome, cystinosis, classic homocystinuria (HCU), Fabry disease, and renal sarcoidosis.

[0121] Optionally, the kidney disease is selected from diabetic nephropathy, focal segmental glomerulosclerosis (FSGS), hypertensive nephrosclerosis, chronic glomerulonephritis, chronic transplant glomerulonephritis, chronic interstitial nephritis, Sjögren's syndrome, Alagille syndrome, α1-antitrypsin deficiency, and polycystic kidney disease.

[0122] The effects of tromethamine salts or crystalline forms on the disease, as defined herein, can be assessed, for example, by measuring eGFR or ACR. Treatment efficacy can be an improvement in eGFR and / or ACR. Treatment efficacy can be stabilization of eGFR and / or ACR. Treatment efficacy can also be a delay or slowing of disease progression, as assessed, for example, by eGFR and / or ACR measurements.

[0123] Optionally and not exhaustively, other therapeutic agents to be used in combination for the treatment of kidney disease are selected from the following: - Endothelin receptor antagonists (ERAs), such as those selected from bosentan, masitentan, tezosentan, sitasenstan, ambrisentan, atrasentan, BQ-123, zipotetentan, edonantan, A192621 (CAS No. 195529-54-5) and BQ-788 (CAS No. 156161-89-6); - ACE inhibitors (angiotensin-converting enzyme), such as those selected from benazepril, captopril, enalapril, fosinopril, lisinopril, moxipril, perindopril, quinapril, ramipril, and quindopril; - ARB (angiotensin II receptor blocker), such as losartan, candesartan, valsartan, irbesartan, telmisartan, eprosartan, olmesartan, azilsartan, and femasartan; - RASS (renin-angiotensin-aldosterone system) antagonists, such as those selected from captopril, imidapril, zolfenpril, candesartan, delapril, telmisartan, aliskiren, moxipril, enalapril, valsartan, fosinopril, irbesartan, perindopril, quinapril, ramipril, iprosartan, olmesartan, trandopril, losartan, azilsartan, lisinopril, spiropril, benazepril, and cilazapril; - β-blockers (β-adrenergic receptor blockers), such as metoprolol, atenolol, bisoprolol, nebivolol, propranolol, nadolol, carvedilol, labetalol, and timolol; - Diuretics, such as those selected from furosemide, azosemide, bumetanide, pyrrolizidine, torasemide ethacrynic acid (torasemide) and ethoxyzoline; - MRA (mineralocorticoid receptor antagonist or aldosterone receptor antagonist), such as spironolactone, eplerenone, canrenone, fenerenone and methylprednisolone propionate; - SGLT2 (sodium-glucose coupled transporter 2) protein inhibitors, also known as glibenclamide, such as betaglibenclamide, canaglibenclamide, dapaglibenclamide, empaglibenclamide, eogglibenclamide, ilaglibenclamide, regagglibenclamide, seragglibenclamide, sogglibenclamide and togglibenclamide; - GLP1 agonists (glucagon-like peptide-1 receptor agonists), such as exenatide, liraglutide, abiglutide, duraglutide, liximab, smegglutide and telpoglycinide. - ELX-02 (CAS No.: 1375073-95-2); and - Peritibacterium (CAS No.: 2304692-47-3).

[0124] More specifically, the additional therapeutic agents to be used in combination to treat CKD or renal fibrosis are selected from the group detailed above, including ERA, ACE inhibitors, ARB, RASS antagonists, β-blockers, diuretics, MRA, SGLT2 inhibitors, GLP1 agonists, and any combination thereof.

[0125] More specifically, another therapeutic agent to be used in combination for the treatment of hereditary kidney diseases such as Albert syndrome cystinosis is ELX-02 (CAS No.: 1375073-95-2).

[0126] More specifically, the additional therapeutic agent to be used in combination for the treatment of HCU is pertibactam (CAS No.: 2304692-47-3).

[0127] Thirdly, the disease is liver disease, especially chronic liver disease. Chronic liver diseases can be selected from primary biliary cirrhosis or primary biliary cholangitis (PBC), brain tendon xanthomas (CTX), primary sclerosing cholangitis (PSC), drug-induced cholestasis, intrahepatic cholestasis of pregnancy, parenteral nutrition-associated cholestasis (PNAC), bacterial overgrowth or sepsis-associated cholestasis, autoimmune hepatitis, chronic viral hepatitis, alcoholic liver disease, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), alcoholic hepatitis, liver transplantation-associated graft-versus-host disease, liver regeneration after living donor liver transplantation, congenital liver fibrosis, choledocholithiasis, granulomatous liver disease, intrahepatic or extrahepatic malignancies, Sjögren's syndrome, Alagille syndrome, sarcoidosis, Wilson's disease, Gaucher disease, hemochromatosis, biliary atresia, liver transplant rejection due to bile duct absence, cystic fibrotic liver disease, and α1-antitrypsin deficiency. In a very specific sense, the disease is NASH.

[0128] Optionally, additional therapeutic agents to be used in combination to treat liver disease, particularly in subjects with NASH, may be: - SGLT2 (sodium-glucose coupled transporter 2) protein inhibitors, also known as glibenclamide, such as betaglibenclamide, canaglibenclamide, dapaglibenclamide, empaglibenclamide, eogglibenclamide, ilaglibenclamide, regagglibenclamide, seragglibenclamide, sogglibenclamide and togglibenclamide; - GLP1 agonists (glucagon-like peptide-1 receptor agonists), such as exenatide, liraglutide, abiglutide, duraglutide, liximab, smegglutide and telpoglycinide. - FGF19 (fibroblast growth factor 19); - FGF21 (fibroblast growth factor 21); - DPP-4 inhibitors (dipeptidyl peptidase 4), such as those selected from sitagliptin, vildagliptin, saxagliptin, linagliptin, giglitazone, alagliptin, ticagliptin, alogliptin, treagliptin, ocagliptin, edagliptin, and gogliptin; - PPAR agonists, such as those selected from nagrieza, erafibrano, lanilano, salrogrieza, and pioglitazone; - SGLT1 (sodium-glucose coupled transporter) inhibitors, such as those selected from soragliflozin, linagliflozin and mizagliflozin; - THR (thyroid hormone receptor) beta agonists such as remdesivir; - FASN (fatty acid synthase) inhibitors such as orlistat; or - Inhibitors of HSD17b13 (hydroxysteroid 17β-dehydrogenase 13), particularly oligonucleotides such as siRNA or antisense oligonucleotides or antibodies that reduce its expression (US-2019106749, the contents of which are incorporated herein by reference).

[0129] In the fourth aspect, the disease is an autoimmune or inflammatory disease. For example, the disease can be selected from multiple sclerosis, rheumatoid arthritis, Behçet's disease, Churg-Strauss syndrome, Guillain-Barré syndrome, bile acid diarrhea (BAD), inflammatory bowel disease (IBD) (including ulcerative colitis and Crohn's disease), and irritable bowel syndrome (IBS), especially bile acid diarrhea (BAD), inflammatory bowel disease (IBD) (including ulcerative colitis and Crohn's disease), and irritable bowel syndrome (IBS).

[0130] In another aspect, the diseases are cardiovascular diseases, such as insulin resistance, type I and type II diabetes, and obesity.

[0131] In another aspect, the disease is cancer. For example, cancer can be a solid tumor or a hematopoietic tumor, preferably selected from AIDS-related Kaposi's sarcoma, leukemias (such as hairy cell leukemia, chronic myeloid leukemia and non-Hodgkin's leukemia), lymphomas (such as follicular lymphoma, cutaneous T-cell lymphoma and adult T-cell leukemia-lymphoma), carcinoid tumors, melanomas, multiple myeloma, renal cell carcinoma, colorectal adenocarcinoma, liver cancer, breast cancer, prostate cancer, ovarian cancer, pancreatic cancer, peritoneal cancer, bladder cancer, lung cancer, glioblastoma, oral cancer, glioma, head and neck cancer, sarcoma, and neuroendocrine tumors. Other therapeutic agents can be any drug that can be used to treat cancer, non-exhaustively, including chemotherapeutic agents (including inhibitors of topoisomerase I or II, DNA cross-linking agents, DNA alkylating agents, antimetabolites, and inhibitors of the mitotic spindle), radiation therapy, hormone therapy, targeted therapy, such as with HDAC inhibitors, PARP inhibitors, kinase inhibitors (e.g., inhibitors of EGFR, ALK, and KRAS), anti-angiogenic agents, hypomethylating agents, cancer vaccines, checkpoint inhibitors, immunotherapy, T-cell-based cancer immunotherapy (including adoptive cell transfer (ACT), genetically modified T cells, or engineered T cells such as chimeric antigen receptor cells (CAR-T cells)), and antibody-drug conjugates.

[0132] In another aspect, the diseases can be selected from age-related macular degeneration, angiomatosis, thrombocytosis, polycythemia vera, idiopathic extramedullary metaplasia, leishmaniasis, osteoporosis, and chronic fatigue syndrome.

[0133] Other aspects and advantages of the invention will be described in the following embodiments, which should be considered illustrative rather than limiting.

[0134] Example Example 1 Synthesis of EYP001 aminobutadiene triol salt 1. Synthesis Sample A: Add the stoichiometric amount of tromethamine (dissolved in EtOH / H2O 2 / 0.5 mL) corresponding to a 1:1 ratio to a solution of 230 mg EYP001 dissolved in THF (5 mL).

[0135] Then, the solvent and water are removed under vacuum (40°C), resulting in a strong solid precipitate on the flask wall.

[0136] Add ethanol (up to 4 mL) and stir at 40°C to resuspend the solid.

[0137] Add water (up to 2 mL) and stir at 40°C to redissolve the solid. Then keep the solution at 4°C overnight, which leads to strong crystallization.

[0138] The supernatant was then removed from the crystals, and the crystals were then vacuum dried at 60°C for about 2 hours.

[0139] Sample B: Add a stoichiometric amount of tromethamine (dissolved in EtOH / H2O 1 / 1 mL) to a solution containing 400 mg EYP001 dissolved in THF (10 mL).

[0140] Then, the solvent and water are removed under vacuum (40°C), resulting in a strong solid precipitate on the flask wall.

[0141] Add ethanol (up to 9 mL) and stir at 40°C to resuspend the solid.

[0142] Add water (up to 3.75 mL) and stir at 50°C for about 1.5 hours to allow the solid to redissolve. Then keep the solution at 4°C overnight, which will result in strong crystallization.

[0143] The solvent was then removed from the crystals by filtration, and the crystals were then vacuum dried at 60°C for about 2 hours.

[0144] 2. Analysis of the products X-ray powder diffraction (XRPD) X-ray diffraction (XRD) analysis was performed using a copper reverse cathode, a single-crystal silicon sample holder, and a LynxEye linear detector on a Brüker D2 Phaser diffractometer. The instrument operating conditions for X-ray pattern acquisition were as follows:

[0145] The powder sample is mounted on a flat single-crystal silicon sample holder in a manner that avoids preferred orientation (particles are not randomly oriented) and ensures the planarity of the sample surface.

[0146] Figure 1 XRPD plots of EYP001 tromethamine salt (sample A), tromethamine counterion, and EYP001 are presented. These plots clearly show the separation of a new crystalline form (form I). Table 1 below discloses the corresponding... Figure 2 A list of XRPD spectra.

[0147] Table 1:

[0148] Differential scanning calorimetry (DSC) Differential scanning calorimetry (DSC) analysis was performed on a Q2000 TA Instruments analyzer. The sample to be analyzed was weighed in an aluminum box, then rolled up and placed in the calorimeter oven.

[0149] The instrument operating conditions for DSC spectrum acquisition are as follows:

[0150] Figure 3 Table 4 presents the DSC spectrum of EYP001 aminobutane triol salt (sample A) and the detailed DSC integration results.

[0151] Several events occurred during heating prior to the main thermal decomposition of the compound (observed at an initial temperature of 234 °C).

[0152] • A wide range of endothermic temperatures with onset / peak temperatures of 77°C / 96°C was detected, which may be attributed to the removal of residual water; • A small exothermic reaction was detected with an onset / peak temperature of 147°C / 157°C, which may be attributed to a solid-solid transition and likely indicates a polymorphic change.

[0153] Table 4: Quantitative DSC results of EYP001 tromethamine salt (sample A)

[0154] Thermogravimetric Analysis (TGA) Thermogravimetric analysis was performed on a TA instrument TGA Hi-Res 2950 equipped with an escape gas analysis furnace. The sample was placed in an open aluminum basket and analyzed under the following conditions:

[0155] Figure 4 Table 5 presents the TGA spectrum and detailed TGA integration results of tromethamine salt (sample A).

[0156] Upon heating, three mass losses were detected before the compound began thermal decomposition (observed at an initial temperature of 217 °C), which can be attributed to the removal of residual solvent / water.

[0157] - The first mass loss was 0.2%, and the detected start / end temperature was 20 / 22℃; - The second mass loss was 0.9%, and the detected start / end temperature was 47 / 63℃; - The third mass loss was 2.2%, and the detected start / end temperature was 151 / 182℃.

[0158] Table 5: Quantitative TGA results of EYP001 tromethamine salt (sample A)

[0159] Dynamic vapor adsorption (DVS) Dynamic vapor adsorption (DVS) analysis (gas-water adsorption) was performed on a DVS-Intrinsic incubator (SMS Ltd) equipped with DVS-Intrinsic control software 1.0. Samples placed in an aluminum basket were subjected to full-cycle analysis (adsorption-desorption cycle) under the following conditions:

[0160] First, pre-dry the sample under a stream of dry air until a stable sample mass is observed. Then, increase the relative humidity in 5% increments. At each step, allow the sample mass to increase until equilibrium is reached (based on a given dm / dt target value), and then increase the relative humidity again. Raise the relative humidity to 95%. After equilibrium is reached at this point, begin desorption in a similar stepwise manner, where the sample mass is again reduced to a stable level after each incremental decrease in humidity.

[0161] Through the two steps of the above method, the sample quality record allows for a description of the relationship between the entire vapor water adsorption / desorption behavior and relative humidity.

[0162] Sample EYP001 tromethamine (sample A) was dried (maintained at 0% RH) before undergoing the adsorption-desorption cycle. During the pre-drying phase (the sample was maintained at 25°C / 0% RH before the start of the vapor-water adsorption process), the sample lost 0.9% of its initial mass. This mass loss is approximately consistent with the first mass loss observed by TGA.

[0163] Figure 5 The DVS isotherm plot (water adsorption / desorption trace) recorded for the tested sample is shown.

[0164] During adsorption, the water absorption rate increases systematically, reaching 2.7% at 25℃ / 95% RH.

[0165] The desorption process confirmed that the loss of adsorbed water was quite linear and regular.

[0166] When returning to 0% RH after a complete adsorption-desorption cycle, the final sample mass is almost the same as the initial sample mass (at the start of the first 25℃ / 0% RH step).

[0167] The water absorption rate at 25°C / 60% RH was 1.3%, and the studied bulk can be considered non-hygroscopic (API bulk is usually described as starting to be hygroscopic when the mass increase exceeds 2% at 25°C / 60% RH).

[0168] Polymorph screening EYP001 Amorphous form of tromethamine salt The amorphous material was prepared by freeze-drying of EYP001 tromethamine salt (Form I).

[0169] EYP001 tromethamine salt (form I) was dissolved in THF / water (1:9 v:v) at a concentration between 50 and 70 mg / mL. The solution was then freeze-dried to form amorphous tromethamine salt.

[0170] Figure 16 XRPD images of the amorphous form of EYP001 tromethamine salt and crystalline forms I, II, and III of EYP001 tromethamine salt are presented. These images clearly show the separation of the amorphous form of EYP001 tromethamine salt.

[0171] EYP001 Aminobutane Triol Salt Crystallization Form II Starting with crystalline form I, a supplementary crystalline form II was obtained.

[0172] Add 5 mL of THF to 200 mg of EYP001 tromethamine salt form I. Stir the white suspension overnight at 50 °C, then filter. Wash the separated solid with the saturated filtrate and dry under vacuum at 50 °C for 1 hour.

[0173] X-ray powder diffraction (XRPD) X-ray diffraction (XRD) analysis was performed using a copper reverse cathode, a single-crystal silicon sample holder, and a LynxEye linear detector on a Brüker D2 Phaser diffractometer. The instrument operating conditions for X-ray pattern acquisition were as follows:

[0174] The powder sample is mounted on a flat single-crystal silicon sample holder in a manner that avoids preferred orientation (particles are not randomly oriented) and ensures the planarity of the sample surface.

[0175] Figure 6 XRPD plots of EYP001 tromethamine form II and EYP001 tromethamine form I are presented. These plots visually clearly show the separation of the new crystalline form (form II). Table 2 below discloses the corresponding... Figure 7 List of XRPD spectra.

[0176] Table 2:

[0177] Differential scanning calorimetry (DSC) Differential scanning calorimetry (DSC) analysis was performed on a Q2000 TA Instruments analyzer. The sample to be analyzed was weighed in an aluminum box, then rolled up and placed in the calorimeter oven.

[0178] The instrument operating conditions for DSC spectrum acquisition are as follows:

[0179] Figure 8 Table 6 presents the DSC spectrum of EYP001 aminobutadiene triol salt (form II) and the detailed DSC integration results.

[0180] During heating, an endothermic event with an initial temperature of 236 °C was observed and may be associated with the accompanying melting / decomposition of the salt polymorph.

[0181] Table 6: Quantitative DSC results of EYP001 tromethamine salt (form II)

[0182] Thermogravimetric Analysis (TGA) Thermogravimetric analysis was performed on a TA instrument TGA Hi-Res 2950 equipped with an escape gas analysis furnace. The sample was placed in an open aluminum basket and analyzed under the following conditions:

[0183] Figure 9 Table 7 presents the TGA spectrum and detailed TGA integration results of tromethamine (form II).

[0184] No mass loss was detected before the main thermal decomposition of the compound (observed at an onset temperature of 220 °C) began upon heating.

[0185] Table 7: Quantitative TGA results of EYP001 tromethamine salt (form II)

[0186] Dynamic vapor adsorption (DVS) Dynamic vapor adsorption (DVS) analysis (gas-water adsorption) was performed on a DVS-Intrinsic incubator (SMS Ltd) equipped with DVS-Intrinsic control software 1.0. Samples placed in an aluminum basket were subjected to full-cycle analysis (adsorption-desorption cycle) under the following conditions:

[0187] First, pre-dry the sample under a stream of dry air until a stable sample mass is observed. Then, increase the relative humidity in 5% increments. At each step, allow the sample mass to increase until equilibrium is reached (based on a given dm / dt target value), and then increase the relative humidity again. Raise the relative humidity to 95%. After equilibrium is reached at this point, begin desorption in a similar stepwise manner, where the sample mass is again reduced to a stable level after each incremental decrease in humidity.

[0188] Through the two steps of the above method, the sample quality record allows for a description of the relationship between the entire vapor water adsorption / desorption behavior and relative humidity.

[0189] The EYP001 tromethamine salt form II sample was dried (maintained at 0% RH) before undergoing the adsorption-desorption cycle. No significant mass loss was observed during the pre-drying phase (the sample was maintained at 25°C / 0% RH before initiating the steam-water adsorption process).

[0190] Figure 10 The DVS isotherm plot (water adsorption / desorption trace) recorded for the tested sample is shown.

[0191] During adsorption, very slight changes in sample mass were observed, with a maximum water absorption rate of 0.9% at 25°C / 95% RH.

[0192] Desorption and adsorption spectra Figure 1 To.

[0193] When returning to 0% RH after a complete adsorption-desorption cycle, the final sample mass is almost the same as the initial sample mass (at the start of the first 25℃ / 0% RH step).

[0194] With a water absorption rate of only 0.3% at 60% RH, the studied bulk material can be considered non-hygroscopic (API bulk materials are usually described as starting to be hygroscopic when the mass increase exceeds 2% at 25°C / 60% RH).

[0195] EYP001 Tromethamine Salt Crystallization Form III Starting from the amorphous form, the supplementary crystalline form III was obtained.

[0196] CH3CN was added to the amorphous form of EYP001 tromethamine to obtain a suspension, which was then aged at 4°C for several hours. The solid was then separated by filtration, washed with saturated filtrate, and dried at room temperature for several days.

[0197] X-ray powder diffraction (XRPD) X-ray diffraction (XRD) analysis was performed using a copper reverse cathode, a single-crystal silicon sample holder, and a LynxEye linear detector on a Brüker D2 Phaser diffractometer. The instrument operating conditions for X-ray pattern acquisition were as follows:

[0198] The powder sample is mounted on a flat single-crystal silicon sample holder in a manner that avoids preferred orientation (particles are not randomly oriented) and ensures the planarity of the sample surface.

[0199] Figure 11 XRPD plots of EYP001 tromethamine form III and EYP001 tromethamine form I are presented. These plots visually clearly show the separation of the new crystalline form (form III). Table 3 below discloses the corresponding... Figure 12 List of XRPD spectra.

[0200] Table 3:

[0201] Differential scanning calorimetry (DSC) Differential scanning calorimetry (DSC) analysis was performed on a Q2000 TA Instruments analyzer. The sample to be analyzed was weighed in an aluminum box, then rolled up and placed in the calorimeter oven.

[0202] The instrument operating conditions for DSC spectrum acquisition are as follows:

[0203] Figure 13 Table 8 presents the DSC spectrum of EYP001 aminobutadiene triol salt (form III) and the detailed DSC integration results.

[0204] Several events occurred during heating prior to the accompanying melting / decomposition of the salt polymorph (observed at an initial temperature of 237 °C).

[0205] - The first broad endothermic period starts at 29°C, which may be associated with the removal of the adsorbed solvent / water; - A second broad endothermic phase, followed by a small broad exothermic phase, which together may be attributed to melting / recrystallization and / or solid-solid transitions in the bulk.

[0206] Table 8: Quantitative DSC results of EYP001 tromethamine salt (form III)

[0207] Thermogravimetric Analysis (TGA) Thermogravimetric analysis was performed on a TA instrument TGA Hi-Res 2950 equipped with an escape gas analysis furnace. The sample was placed in an open aluminum basket and analyzed under the following conditions:

[0208] Figure 14 Table 9 presents the TGA spectrum and detailed TGA integration results for tromethamine (form III).

[0209] Upon heating, a 3.0% mass loss was detected before the main thermal decomposition of the compound (observed at an onset temperature of 218 °C) began, at onset / end temperature of 23 / 35 °C, and could be attributed to the removal of solvent / water.

[0210] Table 9: Quantitative TGA results of EYP001 tromethamine salt (form III)

[0211] Dynamic vapor adsorption (DVS) Dynamic vapor adsorption (DVS) analysis (gas-water adsorption) was performed on a DVS-Intrinsic incubator (SMS Ltd) equipped with DVS-Intrinsic control software 1.0. Samples placed in an aluminum basket were subjected to full-cycle analysis (adsorption-desorption cycle) under the following conditions:

[0212] First, pre-dry the sample under a stream of dry air until a stable sample mass is observed. Then, increase the relative humidity in 5% increments. At each step, allow the sample mass to increase until equilibrium is reached (based on a given dm / dt target value), and then increase the relative humidity again. Raise the relative humidity to 95%. After equilibrium is reached at this point, begin desorption in a similar stepwise manner, where the sample mass is again reduced to a stable level after each incremental decrease in humidity.

[0213] Through the two steps of the above method, the sample quality record allows for a description of the relationship between the entire vapor water adsorption / desorption behavior and relative humidity.

[0214] The EYP001 tromethamine salt form III sample was dried (maintained at 0% RH) before undergoing the adsorption-desorption cycle. During the pre-drying phase (the sample was maintained at 25°C / 0% RH before the start of the vapor-water adsorption process), the sample lost approximately 4.6% of its initial mass.

[0215] Figure 15The DVS isotherm plot (water adsorption / desorption trace) recorded for the tested sample is shown.

[0216] During adsorption, the water absorption rate increases rapidly at 15-20% RH, with a mass increase of approximately 4%. Then, the water absorption rate increases more regularly to 7.5% by weight at 25°C / 95% RH.

[0217] Desorption follows a similar spectrum, with a maximum hysteresis of 0.9% at 60% RH.

[0218] When returning to 0% RH after a complete adsorption-desorption cycle, the final sample mass is almost the same as the initial sample mass (at the start of the first 25℃ / 0% RH step).

[0219] Based on the mass lost during the drying step (which is rapidly recovered before reaching 25°C / 15% RH), the bulk can be considered to exist in a monohydrated form (the theoretical percentage of monohydrate in the salt is 2.9%). This monohydrate also regularly adsorbs water before reaching 95% RH.

[0220] EYP001 Aminobutane Triol Salt Crystallization Form IV The recrystallized form IV was obtained by grinding 10 mg of EYP001 tromethamine salt in 0.1 mL of water and aging it at 50 °C.

[0221] Table 10 below lists the XRPD diagrams for Form IV.

[0222] Table 10: Form IV

[0223] Example 2: Solubility Study Materials and methods The solubility kinetics of the salt form and the native EYP001 free acid form were determined in FaSSIF and FeSSIF media, as well as in acetate buffer at pH 4.5, as follows:

[0224] Samples were prepared by adding an excess of each test substance (salt or natural EYP001) to a given volume of test medium. The suspension was stirred by orbital agitation at 37°C in the dark for 1 hour (in acetate buffer at pH 4.5) or 24 hours (in FaSSIF and FeSSIF media).

[0225] The soluble concentration was assessed in an acetate buffer solution at pH 4.5 after stirring for 0.5 hours and 1 hour.

[0226] In FaSSIF medium, the soluble concentration was assessed after 24 hours of stirring. In FeSSIF medium, the soluble concentration was assessed after 0.5 hours, 1 hour, 2 hours, and 24 hours of stirring.

[0227] At each time point, the "supernatant" was separated and diluted in a solvent mixture to allow it to be injected into the chromatographic system. The concentration of the solution in the test medium was determined by HPLC using the external standard method.

[0228] result Tables 11-13 below present the solubility kinetics results in acetate buffer, FaSSIF (fasting state simulated intestinal fluid), and FeSSIF (feeding state simulated intestinal fluid) media at pH 4.5.

[0229] Table 11:

[0230] Table 12:

[0231] ND: Not measured Table 13:

[0232] As shown in Table 11, all forms (Form I, Form II, and Form III) of EYP001 tromethamine salts exhibited significantly higher solubility in acetate buffer at pH 4.5 than EYP001 free acid, EYP001 phenethylbenzylamine salt, EYP001 zinc salt, EYP001 benzylamine salt, and EYP001 ethylenediamine salt. In FaSSIF media, all forms (Form I, Form II, and Form III) of EYP001 tromethamine salts also showed higher solubility than EYP001 free acid, EYP001 piperazine salt, EYP001 morpholine salt, EYP001 imidazole salt, EYP001 phenethylbenzylamine salt, EYP001 zinc salt, and EYP001 cyclohexylamine salt (Table 12). In FeSSIF media, all forms (Form I, Form II and Form III) of EYP001 tromethamine salts also showed higher solubility than EYP001 free acid and EYP001 potassium salt (Table 13).

[0233] Example 3 Pharmacokinetic studies Materials and methods In the first study (Experiment #1), EYP001 free acid, EYP001 4,2-aminoethylmorpholine salt, and EYP001 tromethamine salt form I were prepared in an aqueous formulation (0.5% CMC, 0.25% Tween 80, water) at a dose of 50 mg Eq for each compound. EYP001 A dose of / kg was administered orally to three male rats. Blood samples were collected at 0.25, 0.5, 1, 2, 4, 6, 8, 12 and 24 hours after oral administration.

[0234] In the second study (Experiment #2), EYP001 free acid, EYP001 tromethamine salt form II, and EYP001 tromethamine salt form III were prepared in an aqueous formulation (0.5% CMC, 0.25% Tween 80, water) at a dose of 10 mg Eq for each compound. EYP001 The dose of / kg was administered orally to eight male rats. Blood samples were collected at 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 12 h and 24 h after oral administration.

[0235] result The results of the PK study are presented in Tables 14A and 14B below.

[0236] Table 14A:

[0237] SD: Standard Deviation CV%: % of relative standard deviation Table 14B:

[0238] CV%: % of relative standard deviation in conclusion: As described in Table 14A, in Experiment #1, EYP001 tromethamine salt form I showed superior exposure (AUC) compared to EYP001 free acid and EYP001 4,2-aminoethylmorpholine salt. 0-24 ).

[0239] As described in Table 14B, in Experiment #2 using more animals, EYP001 tromethamine salt form II and EYP001 tromethamine salt form III showed superior exposure (AUC) compared to EYP001 free acid. 0-24 Furthermore, this study clearly demonstrates that, compared with the free EYP001 acid, the use of EYP001 tromethamine salts (including crystalline forms II and III) reduces the variability of exposure and Cmax (see CV%) in Table 14B.

Claims

1. Tromethamine salt of 4-chloro-5-[4-(2,6-dichlorophenyl)sulfonyl piperazin-1-yl]-1- benzofuran-2-carboxylic acid.

2. The tromethamine salt of claim 1 in crystalline form.

3. The tromethamine salt of claim 2, wherein the X-ray diffraction pattern of the crystalline form comprises peaks at the following diffraction angles 2-Theta (2q): 15.2° ± 0.2°, 16.0° ± 0.2°, 17.1° ± 0.2°, 18.3° ± 0.2°, 18.7° ± 0.2°, 20.6° ± 0.2°, 22.6° ± 0.2°, and 23.2° ± 0.2°, wherein the X-ray diffraction pattern is obtained with a Cu Kcc anode.

4. The tromethamine salt of claim 2 or 3, wherein the X-ray diffraction pattern of the crystalline form comprises peaks at the following diffraction angles 2-Theta (2q): 6.3° ± 0.2°, 6.9° ± 0.2°, 8.4° ± 0.2°, 9.6° ± 0.2°, 10.0° ± 0.2°, 11.0° ± 0.2°, 12.8° ± 0.2°, 13.9° ± 0.2°, 15.2° ± 0.2°, 15.5° ± 0.2°, 16.0° ± 0.2°, 16.6° ± 0.2°, 17.1° ± 0.2°, 18.3° ± 0.2°, 18.7° ± 0.2°, 19.2° ± 0.2°, 19.9° ± 0.2°, 20.6° ± 0.2°, 21.0° ± 0.2°, 21.8° ± 0.2°, 22.6° ± 0.2°, 23.2° ± 0.2°, 23.6° ± 0.2°, 24.6° ± 0.2°, 25.7° ± 0.2°, 27.1° ± 0.2°, 28.5° ± 0.2°, 29.7° ± 0.2°, and 29.8° ± 0.2°, wherein the X-ray diffraction pattern is obtained with a Cu Kcc anode.

5. The tromethamine salt of any one of claims 2 to 4, wherein the X-ray diffraction pattern of the crystalline form comprises the following peaks: wherein the X-ray diffraction pattern is obtained with a Cu Kcc anode.

6. The tromethamine salt of claim 2, wherein the X-ray diffraction pattern of the crystalline form comprises peaks at the following diffraction angles 2-Theta (2q): 9.0° ± 0.2°, 14.5° ± 0.2°, 15.9° ± 0.2°, 17.9° ± 0.2°, 20.1° ± 0.2°, 21.4° ± 0.2°, and 25.2° ± 0.2°, wherein the X-ray diffraction pattern is obtained with a Cu Kcc anode.

7. The tromethamine salt according to claim 2 or 6, wherein the X-ray diffraction pattern of the crystalline form comprises peaks at the following diffraction angles 2-Theta (2q): 7.0°±0.2°, 9.0°±0.2°, 11.3°±0.2°, 12.3°±0.2°, 14.0°±0.2°, 14.5°±0.2°, 15.9°±0.2°, 16.3°±0.2°, 16.6°±0.2°, 17.9°±0.2°, 18.7°±0.2°, 20.1°±0.2°, 21.4°±0.2°, 22.7°±0.2°, 23.1°±0.2°, 23.8°±0.2°, 24.3°±0.2°, 25.2°±0.2°, 26.4°±0.2, 26.9°±0.2, 29.1°±0.2, 30.4.°±0.2, 31.0°±0.2, 33.6°±0.2, 34.9°±0.2, and 35.8°±0.2°, wherein the X-ray diffraction pattern is obtained with a Cu K a anode.

8. The tromethamine salt according to any one of claims 2, 6 and 7, wherein the X-ray diffraction pattern of the crystalline form comprises the following peaks: wherein the X-ray diffraction pattern is obtained with a Cu K a anode.

9. The tromethamine salt according to claim 2, wherein the X-ray diffraction pattern of the crystalline form comprises peaks at the following diffraction angles 2-Theta (2q): 11.1°±0.2°, 15.7°±0.2°, 15.9°±0.2°, 17.3°±0.2°, 20.9°±0.2°, 21.2°±0.2°, 21.8°±0.2°, and 27.2°±0.2°, wherein the X-ray diffraction pattern is obtained with a Cu K a anode.

10. The tromethamine salt according to claim 2 or 9, wherein the X-ray diffraction pattern of the crystalline form comprises peaks at the following diffraction angles 2-Theta (2q): 6.3°±0.2°, 7.0°±0.2°, 8.7°±0.2°, 10.5°±0.2°, 11.1°±0.2°, 14.7°±0.2°, 15.9°±0.2°, 17.3°±0.2°, 18.3°±0.2°, 20.0°±0.2°, 20.9°±0.2°, 21.8°±0.2°, 22.7°±0.2°, 23.2°±0.2°, 25.1°±0.2°, 25.9°±0.2°, 27.2°±0.2°, 28.3°±0.2°, 29.4°±0.2°, 31.8°±0.2°, 32.4°±0.2°, 35.2°±0.2°, 36.0°±0.2°, and 37.0°±0.2°, wherein the X-ray diffraction pattern is obtained with a Cu K a anode.

11. The tromethamine salt according to any one of claims 2, 9 and 10, wherein the X-ray diffraction pattern of the crystalline form comprises the following peaks: wherein said X-ray diffractogram is obtained with Cu K alpha anode.

12. The tromethamine salt according to any one of claims 1 to 11 for use as a medicament or pharmaceutical product.

13. A pharmaceutical or veterinary composition comprising the tromethamine salt according to any one of claims 1 to 11 and a pharmaceutically acceptable excipient.

14. The pharmaceutical or veterinary composition according to claim 13, wherein the pharmaceutical or veterinary composition further comprises an additional therapeutic agent, such as a TLR3 agonist, a TLR7 agonist, a TLR8 agonist, a TLR9 agonist, a RIG-I modulator, a STING agonist, an antiviral agent such as a Bulevirtide, an antibacterial agent, an interferon or a pegylated version thereof, a checkpoint inhibitor such as a PD-1 or PD-L1 agonist, an ERA, an ACE inhibitor, an ARB, a RASS antagonist, a beta-blocker, a diuretic, an MRA, an SGLT2 inhibitor, a GLP1 agonist, a SGLT1 inhibitor, FGF19, FGF21, a DPP-4 inhibitor, a PPAR agonist, a THR beta agonist, FASN, HSD17b13 inhibitor, or a combination thereof.

15. The pharmaceutical or veterinary composition according to claim 13 or 14 for use in the treatment of a disease selected from the group consisting of chronic liver diseases, gastrointestinal diseases, kidney diseases, cardiovascular diseases, metabolic diseases, infections, cancer and autoimmune diseases.

16. The pharmaceutical or veterinary composition for use according to claim 15, wherein the disease is an infection, in particular a chronic infection, preferably a viral infection, more preferably an infection caused by Hepatitis B virus (HBV), Hepatitis C virus (HCV), Hepatitis D virus (HDV), Herpes Simplex Virus (HSV), Papillomavirus (HPV) (e.g. Condyloma acuminatum), Varicella-Zoster virus, Cytomegalovirus (CMV), Rhinovirus, Hepatitis A virus, Hepatitis E virus, Kaposi’s sarcoma herpesvirus, a Coronavirus (including SARS-CoV1, MERS-CoV and SARS-CoV2), a Retrovirus (including HIV) and an Influenza virus.

17. The pharmaceutical or veterinary composition for use according to claim 16 in combination with a TLR3 agonist, a TLR7 agonist, a TLR8 agonist, a TLR9 agonist, a RIG-I modulator, a STING agonist, an antiviral agent such as a Bulevirtide, an antibacterial agent, an interferon or a pegylated version thereof, a checkpoint inhibitor such as a PD-1 or PD-L1 agonist, or a combination thereof.

18. The pharmaceutical or veterinary composition for use according to claim 15, wherein the disease is a kidney disease, in particular a kidney disease comprising kidney fibrosis and / or chronic kidney disease (CKD), for example selected from the group consisting of hypertension, type 2 diabetes, type 1 diabetes, obesity, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), metabolic (dysfunction)-associated fatty liver disease (MAFLD), aging, infectious glomerulonephritis (in particular infections such as syphilis, malaria, hepatitis B, hepatitis C or HIV), focal segmental glomerulosclerosis, IgA nephropathy, minimal change glomerulopathy, membranous nephropathy, renal vasculitis, urinary obstruction, genetic alterations, autoimmune diseases (such as systemic lupus erythematosus (SLE)) and drug- or toxin-induced kidney disease (such as kidney disease induced by drugs like captopril, NSAIDs, penicillamine, probenecid, bursine, anti-TNF therapies and tiopronin or by toxins like inorganic salts (e.g., gold, mercury)), AIDS-related kidney disease, ischemic kidney disease, tubulointerstitial kidney disease, hepatorenal syndrome, hydronephrosis, kidney dysplasia, medullary cystic kidney disease, medullary sponge kidney, polycystic dysplastic kidney, podocytopathy, renal papillary necrosis, nephritis (including glomerulonephritis, hereditary nephritis, interstitial nephritis, pyelonephritis), nephrocalcinosis, nephrosclerosis, Alport syndrome, cystinosis, classic homocystinuria (HCU), Fabry disease, nephronodular disease, diabetic nephropathy, focal segmental glomerulosclerosis (FSGS), hypertensive nephrosclerosis, chronic glomerulonephritis, chronic transplant glomerulopathy, chronic interstitial nephritis, Sjogren's syndrome, Alagille syndrome, alpha 1-antitrypsin deficiency and polycystic kidney disease.

19. The pharmaceutical or veterinary composition for use according to claim 18, wherein the disease is chronic kidney disease (CKD).

20. The pharmaceutical or veterinary composition for use according to claim 18 or 19, in combination with an ERA, an ACE inhibitor, an ARB, a RASS antagonist, a beta-blocker, a diuretic, an MRA, an SGLT2 inhibitor, a GLP1 agonist or a combination thereof.

21. The pharmaceutical or veterinary composition for use according to claim 15, wherein the disease is a liver disease, in particular a chronic liver disease, preferably primary biliary cirrhosis or primary biliary cholangitis (PBC), cerebral tendon xanthomatosis (CTX), primary sclerosing cholangitis (PSC), drug-induced cholestasis, intrahepatic cholestasis of pregnancy, parenteral nutrition-associated cholestasis (PNAC), bacterial overgrowth or sepsis-associated cholestasis, autoimmune hepatitis, chronic viral hepatitis, alcoholic liver disease, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), alcoholic hepatitis, liver transplant-related graft-versus-host disease, liver regeneration after living donor liver transplantation, congenital liver fibrosis, choledocholithiasis, granulomatous liver disease, intra- or extrahepatic malignancy, Sjogren syndrome, Alagille syndrome, sarcoidosis, Wilson disease, Gaucher disease, hemochromatosis, biliary atresia, ductopenic liver transplant rejection, cystic fibrosis liver disease and alpha 1-antitrypsin deficiency.

22. The pharmaceutical or veterinary composition for use according to claim 21, in combination with an SGLT2 inhibitor, a GLP1 agonist, an SGLT1 inhibitor, FGF19, FGF21, a DPP-4 inhibitor, a PPAR agonist, a THR beta agonist, FASN, HSD17b13 inhibitor or a combination thereof.

23. A method of treating a disease selected from chronic liver disease, gastrointestinal disease, kidney disease, cardiovascular disease, metabolic disease, infection, cancer and autoimmune disease in a subject in need thereof, the method comprising administering to the subject an effective amount of a pharmaceutical or veterinary composition according to claim 13 or 14.

24. Use of a pharmaceutical or veterinary composition according to claim 13 or 14 for the manufacture of a medicament for the treatment of a disease selected from chronic liver disease, gastrointestinal disease, kidney disease, cardiovascular disease, metabolic disease, infection, cancer and autoimmune disease.

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