This specification relates to certain amino heteroaromatic compounds and pharmaceutically acceptable salts thereof that inhibit 17P hydroxy steroid dehydrogenase 13 (17PHSD13 or HSD17B13), and their use in treating diseases such as liver disease. This specification also relates to processes and intermediate compounds involved in the preparation of the amino heteroaromatic compounds and to pharmaceutical compositions containing them.

Introduction

Non-alcoholic fatty liver disease (NAFLD) represents a spectrum of liver disease ranging from simple steatosis (non-alcoholic fatty liver), to non-alcoholic steatohepatitis (NASH) with or without fibrosis, to cirrhosis. Hepatic steatosis is defined as excess fat accumulation in the liver with greater than 5% induced by causes other than alcohol intake. NASH is defined by hepatic steatosis with inflammation and hepatocyte injury, with or without fibrosis. It is estimated that approximately 25% of the global population has NAFLD, and mortality due to NAFLD-related disease is expected to increase significantly through 2030.

To date, there are no approved treatments for NAFLD (such as NASH) and therapeutic interventions focus on addressing co-morbidities that contribute to the pathogenesis of NAFLD, including treating insulin resistance, obesity, type II diabetes mellitus, and dyslipidemia.

Recently, a variant in the 17PHSD13 gene, was associated in an allele dose-dependent manner with decreased serum aminotransferases levels, as well as a lower risk of liver disease, including alcoholic and non-alcoholic liver disease, cirrhosis and hepatocellular carcinoma (HCC) (Abul-Husn et al, N Engl J Med. 2018, 378(12), 1096-106, Wang et al, Eur Rev Med Pharmacol Sci, 2020, 24(17), 8997-9007). The 17PHSD13 splice variant (rs72613567:TA) results in a truncated, unstable and enzymatically inactive protein and has thus been characterized as an 17PHSD13 Loss of Function (LoF) variant (Ma et al, Hepatology 2019, 69(4), 1504-19). The association between the LoF 17PHSD13 (rs72613567:TA) and decreased disease severity has been replicated in additional cohorts with histologically proven NAFLD and was also associated with lower plasma transaminases, reduced risk of cirrhosis, HCC and liver related mortality in a study of 111612 individuals from the Danish general population (Gellert-Kristensen et al, Hepatology, 2020, 71(1), 56-66). Interestingly, the protective effect of the LoF 17PHSD13 (rs72613567:TA) variant on plasma transaminases levels appears to be amplified by several key risk factors of liver disease such as obesity, alcohol consumption, as well as established genetic risk factors such as, but not limited to, the (rs738409 C>G) variant in patatin-like phospholipase domain-containing protein 3 (PNPLA3). Further, two additional 17PHSD13LoF variants (rs62305723) and (rsl43404524) were also reported to confer protection from chronic liver disease progression (Kozlitina et al, N Engl J Med, 2018, 379(19), 1876-7). In general, the LoF 17PHSD13 protective variants has a stronger association with fibrosis and progression to advance liver disease but is not associated with steatosis.

Based on the genetic validation of 17PHSD13LoF variants conferring protection against liver disease risk and progression, inhibition of 17PHSD13 activity with small molecules inhibitors could be an effective therapeutic approach for treating liver diseases such as NAFLD (for example NASH, liver fibrosis, cirrhosis and isolated steatosis), liver inflammation, alcoholic steatohepatitis (ASH), hepatitis C virus (HCV) and hepatocellular carcinoma (HCC), such as in individuals harbouring several key risk factors of liver disease such as obesity, alcohol consumption, as well as established genetic risk factors such as the (rs738409 C>G) variant in PNPLA3.

The compounds of the disclosure provide an anti-liver disease effect by, as a minimum, acting as 17PHSD13 inhibitors. Further, compounds of the disclosure may selectively inhibit 17PHSD13 over 17PHSD4 and/or 17PHSD9.

Fifteen 17PHSD (HSD17B) members have been identified in human. The sequence homology among the different members is rather low, but the overall structure seems conserved. 17P-Hydroxysteroid dehydrogenases are mainly involved in sex hormone metabolism. Some 17PHSD enzymes also play key roles in cholesterol and fatty acid metabolism (Labrie et al. Journal of Molecular Endocrinology, 2000, 25, 1-16, Wen Su et al. Molecular and Cellular Endocrinology, 2019, 489, 119-125). A clean off-target profile is an advantage for a 17PHSD13 inhibitor to avoid potential toxicity caused by off- target activity. This includes selectivity to other 17PHSD members.

17PHSD4/ D-bifunctional protein (DBP) is involved in fatty acid p-oxidation and steroid metabolism. 17PHSD4 is ubiquitously expressed and play an important role in the inactivation of estrogens in a large series of peripheral tissues. Mutations in 17PHSD4 are known to cause DBP deficiency, an autosomal-recessive disorder of peroxisomal fatty acid p-oxidation that is generally fatal within the first two years of life. A homozygous missense variant in 17PHSD4 has been identified in Perrault syndrome, a recessive disorder characterized by ovarian dysgenesis in females, sensorineural deafness in both males and females, and in some patients, neurological manifestations (Pierce et al. Am. J. Hum. Genet., 2010, 87, 282-8; and Chen et al. BMC Med Genet., 2017, 18, 91).

17PHSD9/ RDH5 (retinol dehydrogenase 5) is involved in retinoid metabolism. The enzyme is mainly expressed in the retinal pigment epithelium. The RDH5 gene encodes the enzyme that is a part of the visual cycle, the 11-cis retinol dehydrogenase, catalysing the reduction of 11-cis-retinol to 11-cis- retinal. RDH5 gene mutations cause a progressive cone dystrophy or macular dystrophy as well as night blindness. Fundus albipunctatus is a rare, congenital form of night blindness with rod system impairment, characterised by the presence of numerous small, white-yellow retinal lesions. This disorder is caused mostly by mutations in the RDH5 gene (Hotta et al. Am. J. Ophthalmol., 2003, 135, 917-9; and Skorczyk-Werner et al. J. Appl. Genet., 2015, 56, 317-27).

The compounds of the specification may also exhibit advantageous physical properties (for example, lower lipophilicity, higher aqueous solubility, higher permeability, lower plasma protein binding, and/or greater chemical stability), and/or favourable toxicity profiles (for example a decreased activity at hERG), and/or favourable metabolic or pharmacokinetic profiles, in comparison with other known 17PHSD13 inhibitors. Such compounds may therefore be especially suitable as therapeutic agents, such as for the treatment of liver disease.

General Description

According to one aspect of the specification there is provided a compound, or a pharmaceutically acceptable salt thereof, wherein the compound is 2-(difluoromethoxy)-5-(5-((5-fluoroadamantan-2- yl)(methyl)amino)-l,3,4-oxadiazol-2-yl)phenol or 5-(3-(((l-cyclohexyl-lH-tetrazol-5- yl)methyl)(methyl)amino)-l,2,4-oxadiazol-5-yl)-2-fluoropheno l.

In a further aspect there is provided a pharmaceutical composition comprising a compound of the disclosure or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

In a further aspect there is provided a compound of the disclosure or a pharmaceutically acceptable salt thereof, for use in therapy.

In a further aspect there is provided a compound of the disclosure or a pharmaceutically acceptable salt thereof, for use in the treatment of liver disease.

In a further aspect there is provided the use of a compound of the disclosure or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament.

In a further aspect there is provided the use of a compound of the disclosure or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of liver disease.

In a further aspect there is provided a method of treating liver disease in a patient comprising administering to the patient an effective amount of a compound of the disclosure or a pharmaceutically acceptable salt thereof. Units, prefixes, and symbols are denoted in their International System of Units (SI) accepted form. Numeric ranges are inclusive of the numbers defining the range.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure is related. For example, the Concise Dictionary of Biomedicine and Molecular Biology, Juo, Pei-Show, 2nd ed., 2002, CRC Press; The Dictionary of Cell and Molecular Biology, 3rd ed., 1999, Academic Press; and the Oxford Dictionary of Biochemistry and Molecular Biology, Revised, 2000, Oxford University Press, provide one of skill with a general dictionary of many of the terms used in this disclosure.

Detailed Description

In one aspect there is provided a compound of the disclosure, or a pharmaceutically acceptable salt thereof, as defined above.

In embodiments, the is provided 2-(difluoromethoxy)-5-(5-((5-fluoroadamantan-2- yl)(methyl)amino)-l,3,4-oxadiazol-2-yl)phenol, or a pharmaceutically acceptable salt thereof.

In embodiments, the is provided 5-(3-(((l-cyclohexyl-lH-tetrazol-5-yl)methyl)(methyl)amino)- l,2,4- oxadiazol-5-yl)-2-fluorophenol, or a pharmaceutically acceptable salt thereof.

The compounds of the disclosure, and pharmaceutically acceptable salts thereof, may be prepared, used or supplied in amorphous form, crystalline form, or semicrystalline form and any given compound of the disclosure, or pharmaceutically acceptable salt thereof, may be capable of being formed into more than one crystalline / polymorphic form, including hydrated (e.g. hemi hydrate, a mono hydrate, a di hydrate, a tri hydrate or other stoichiometry of hydrate) and/or solvated forms. It is to be understood that the present specification encompasses any and all such solid forms of the compound of the disclosure, and pharmaceutically acceptable salts thereof.

In further embodiments there is provided a compound of the disclosure, which is obtainable by the methods described in the 'Examples" section hereinafter.

The present specification is intended to include all isotopes of atoms occurring in the present compounds. Isotopes will be understood to include those atoms having the same atomic number but different mass numbers. For example, isotopes of hydrogen include tritium and deuterium. Isotopes of carbon include ¹³ C and ¹⁴ C. Isotopes of nitrogen include ¹⁵ N. Isotopes of fluorine include ¹⁸ F.

A suitable pharmaceutically acceptable salt of a compound of the disclosure is, for example, a base addition salt. A base addition salt of a compound of the disclosure may be formed by bringing the compound into contact with a suitable inorganic or organic base under conditions known to the skilled person. A base addition salt may for example be an alkali metal salt (such as a sodium, potassium, or lithium salt) or an alkaline earth metal salt (such as a calcium salt), which may be formed using an alkali metal or alkaline earth metal hydroxide or alkoxide (e.g., an ethoxide or methoxide). A base addition salt may also be formed using a suitably basic organic amine (e.g., a choline or meglumine salt).

A further suitable pharmaceutically acceptable salt of a compound of the disclosure is, for example, a salt formed within a patient's body after administration of a compound of the disclosure to the patient.

The compound of the disclosure, or pharmaceutically acceptable salt thereof, may be prepared as a co-crystal solid form. It is to be understood that a pharmaceutically acceptable co-crystal of an compound of the disclosure, or pharmaceutically acceptable salts thereof, form an aspect of the present specification.

In a further aspect there is provided a pharmaceutical composition comprising a compound of the disclosure, or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

The term "pharmaceutical composition" refers to a preparation which is in such form as to permit the biological activity of the active ingredient, and which contains no additional components which are unacceptably toxic to a patient to which the composition would be administered. Such compositions can be sterile. A pharmaceutical composition according to the present specification will comprise a compound of the disclosure, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. For example, the composition may be in a form suitable for oral use (for example as tablets, lozenges, hard or soft capsules, aqueous or oily suspensions, emulsions, dispersible powders or granules, syrups or elixirs) or for parenteral administration (for example as a sterile aqueous or oily solution for intravenous, subcutaneous, intramuscular or intramuscular dosing or as a suppository for rectal dosing). Such compositions may be obtained by conventional procedures using conventional pharmaceutical excipients, well known in the art. Thus, compositions intended for oral use may contain, for example, one or more colouring, sweetening, flavouring and/or preservative agents. An effective amount of the compound of the disclosure, or a pharmaceutically acceptable salt thereof, will normally be present in the composition.

A compound of the disclosure, or a pharmaceutically acceptable salt thereof, will normally be administered via the oral route though parenteral, intravenous, intramuscular, subcutaneous or in other injectable ways, buccal, rectal, vaginal, transdermal and/or nasal route and/or via inhalation, in the form of pharmaceutical preparations comprising the active ingredient or a pharmaceutically acceptable salt or solvate thereof, or a solvate of such a salt, in a pharmaceutically acceptable dosage form may be possible. Depending upon the disorder and patient to be treated and the route of administration, the compositions may be administered at varying doses.

The pharmaceutical formulations of the compound of the disclosure, described above may be prepared e.g. for parenteral, subcutaneous, intramuscular or intravenous administration.

The pharmaceutical formulations of the compound of the disclosure, described above may conveniently be administered in unit dosage form and may be prepared by any of the methods well- known in the pharmaceutical art, for example as described in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, PA., (1985).

Pharmaceutical formulations suitable for oral administration may comprise one or more physiologically compatible carriers and/or excipients and may be in solid or liquid form. Tablets and capsules may be prepared with binding agents; fillers; lubricants; and surfactants. Liquid compositions may contain conventional additives such as suspending agents; emulsifying agents; and preservatives Liquid compositions may be encapsulated in, for example, gelatin to provide a unit dosage form. Solid oral dosage forms include tablets, two-piece hard shell capsules and soft elastic gelatin (SEG) capsules. An exemplary oral composition would comprise a compound of the disclosure, and at least one pharmaceutically acceptable excipient filled into a two-piece hard shell capsule or a soft elastic gelatin (SEG) capsule.

As a result of their 17BHSD13 inhibitory activity, the compounds of the disclosure, and pharmaceutically acceptable salts thereof are expected to be useful in therapy, for example in the treatment of diseases or medical conditions mediated at least in part by 17BHSD13, including liver disease, such as NASH.

In one aspect of the present specification there is provided a compound of the disclosure, or a pharmaceutically acceptable salt thereof, for use in therapy.

In one aspect of the present specification there is provided a compound of the disclosure, or a pharmaceutically acceptable salt thereof, for use in the treatment of liver disease. In embodiments, the liver disease is selected from alcoholic liver disease, non-alcoholic liver disease, NAFLD (such as NASH, liver fibrosis, cirrhosis, and isolated steatosis), liver inflammation, alcoholic steatoheptatis (ASH), hepatitis C virus (HCV) and hepatocellular carcinoma (HCC).

The term "therapy" is intended to have its normal meaning of dealing with a disease in order to entirely or partially relieve one, some or all of its symptoms, or to correct or compensate for the underlying pathology. The term "therapy" also includes "prophylaxis" unless there are specific indications to the contrary. The terms "therapeutic" and "therapeutically" should be interpreted in a corresponding manner.

The term "prophylaxis" is intended to have its normal meaning and includes primary prophylaxis to prevent the development of the disease and secondary prophylaxis whereby the disease has already developed and the patient is temporarily or permanently protected against exacerbation or worsening of the disease or the development of new symptoms associated with the disease.

The term "treatment" is used synonymously with "therapy". Similarly the term "treat" can be regarded as "applying therapy" where "therapy" is as defined herein.

In embodiments, there is provided a compound of the disclosure, or a pharmaceutically acceptable salt thereof, for use in providing an inhibitory effect on 17PHSD13.

In embodiments, there is provided a compound of the disclosure, or a pharmaceutically acceptable salt thereof, for use in the treatment of a disease mediated by 17PHSD13, such as liver disease (e.g. NASH).

In embodiments, there is provided a compound of the disclosure, or a pharmaceutically acceptable salt thereof, for use in the treatment of fatty liver disease.

In embodiments, there is provided a compound of the disclosure, or a pharmaceutically acceptable salt thereof, for use in the treatment of nonalcoholic Fatty Liver Disease (NAFLD), such as isolated steatosis, Nonalcoholic Steatohepatitis (NASH), liver fibrosis or cirrhosis. In further embodiments, the liver disease is end stage liver disease.

In embodiments, there is provided a compound of the disclosure, or a pharmaceutically acceptable salt thereof, for use in the treatment of liver disease, such as NASH, wherein the patient is also suffering from or susceptible to one or more conditions selected from the group consisting of obesity, dyslipidemia, insulin resistance, Type 2 diabetes, and renal insufficiency.

In embodiments, there is provided a compound of the disclosure, or a pharmaceutically acceptable salt thereof, for use in the treatment of liver disease, such as NASH, wherein the patient has a body mass index (BMI) of 27 kg/m ² to 40 kg/m ² . In further embodiments, the subject has a BMI of 30 kg/m ² to 39.9 kg/m ² . In further embodiments, the patient has a BMI of at least 40 kg/m ² . In further embodiments, the patient is overweight. In further embodiments, the patient is obese. In embodiments, there is provided a compound of the disclosure, or a pharmaceutically acceptable salt thereof, for use in the treatment of liver disease, such as NASH, wherein the patient is also suffering from or susceptible to dyslipidemia.

In embodiments, there is provided a compound of the disclosure, or a pharmaceutically acceptable salt thereof, for use in the treatment of liver disease, such as NASH, wherein the patient is also suffering from or susceptible to insulin resistance.

In embodiments, there is provided a compound of the disclosure, or a pharmaceutically acceptable salt thereof, for use in the treatment of liver disease, such as NASH, wherein the patient is also suffering from or susceptible to Type 2 diabetes.

In embodiments, there is provided a compound of the disclosure, or a pharmaceutically acceptable salt thereof, for use in the treatment of liver disease, such as NASH, wherein the patient is also suffering from or susceptible to renal insufficiency.

In embodiments, there is provided a compound of the disclosure, or a pharmaceutically acceptable salt thereof, for use in the treatment of liver disease, such as NASH, wherein the patient is also suffering from or susceptible to liver fibrosis. In further embodiments, the patient is (i) suffering from or susceptible to liver fibrosis, and (ii) suffering from or susceptible to one or more conditions selected from the group consisting of obesity, dyslipidemia, insulin resistance, Type 2 diabetes, and renal insufficiency.

In embodiments, there is provided a compound of the disclosure, or a pharmaceutically acceptable salt thereof, for use in the treatment of liver disease, such as NASH, wherein the patient is also suffering from or susceptible to cirrhosis. In further embodiments, the patient is (i) suffering from or susceptible to cirrhosis, and (ii) suffering from or susceptible to one or more conditions selected from the group consisting of obesity, dyslipidemia, insulin resistance, Type 2 diabetes, and renal insufficiency.

In embodiments, there is provided a compound of the disclosure, or a pharmaceutically acceptable salt thereof, for use in the treatment of NAFLD. In further embodiments, the NAFLD is Stage 1 NAFLD. In further embodiments, the NAFLD is Stage 2 NAFLD. In further embodiments, the NAFLD is Stage 3 NAFLD. In further embodiments, the NAFLD is Stage 4 NAFLD. See, e.g., "The Diagnosis and Management of Nonalcoholic Fatty Liver Disease: Practice Guidance From the American Association for the Study of Liver Diseases," Hepatology, Vol. 67, No. 1, 2018.

In embodiments, there is provided a compound of the disclosure, or a pharmaceutically acceptable salt thereof, for use in the treatment of NAFLD, such as NASH. In further embodiments, the patient is obese. In further embodiments, the patient has alcoholic liver disease. In further embodiments, the patient has a genetic risk factor for liver disease, such as the (rs738409 C>G) variant in PNPLA3.

In embodiments, there is provided a compound of the disclosure, or a pharmaceutically acceptable salt thereof, for use in the treatment of NASH. In further embodiments, the NASH is Stage 1 NASH. In further embodiments, the NASH is Stage 2 NASH. In further embodiments, the NASH is Stage 3 NASH. In further embodiments, the NASH is Stage 4 NASH. In further embodiments, the patient is also suffering from or susceptible to one or more conditions selected from obesity, dyslipidemia, insulin resistance, Type 2 diabetes, and renal insufficiency.

In embodiments, there is provided a compound of the disclosure, or a pharmaceutically acceptable salt thereof, for use in the treatment of liver fibrosis. In further embodiments, the liver fibrosis is Stage 3 liver fibrosis. In further embodiments, the patient is also suffering from or susceptible to one or more conditions selected from obesity, dyslipidemia, insulin resistance, Type 2 diabetes, and renal insufficiency.

In embodiments, there is provided a compound of the disclosure, or a pharmaceutically acceptable salt thereof, for use in the treatment of cirrhosis. In further embodiments, the cirrhosis is stage F4 cirrhosis. In further embodiments, the patient is also suffering from or susceptible to one or more conditions selected from obesity, dyslipidemia, insulin resistance, Type 2 diabetes, and renal insufficiency.

In embodiments, there is provided a compound of the disclosure, or a pharmaceutically acceptable salt thereof, for use in the treatment of liver inflammation. In further embodiments, the inflammation is chronic inflammation. In further embodiments, the chronic inflammation is selected from the group consisting of rheumatoid arthritis, osteoarthritis, and Crohn's disease. In further embodiments, the chronic inflammation is rheumatoid arthritis.

In embodiments, there is provided a compound of the disclosure, or a pharmaceutically acceptable salt thereof, for use in the treatment of hepatocellular carcinoma (HCC).

In embodiments, there is provided a compound of the disclosure, or a pharmaceutically acceptable salt thereof, for use in the treatment of alcoholic steatoheptatis (ASH).

In embodiments, there is provided a compound of the disclosure, or a pharmaceutically acceptable salt thereof, for use in the treatment of hepatitis C virus (HCV). In one aspect of the present specification there is provided the use of a compound of the disclosure, or a pharmaceutically acceptable salt thereof, as described herein, in the manufacture of a medicament, such as a medicament for the treatment of disease (e.g. NASH).

In one aspect of the present specification there is provided a method of treating disease, such as NASH, in a patient comprising administering to the patient an effective amount of a compound of the disclosure, or a pharmaceutically acceptable salt thereof.

Terms such as "treating" or "treatment" refer to both (1) therapeutic measures that cure, slow down, lessen symptoms of, and/or halt progression of a diagnosed pathologic condition or disorder and (2) prophylactic or preventative measures that prevent and/or slow the development of a targeted pathologic condition or disorder. Thus, those in need of treatment include those already with the disorder; those prone to have the disorder; and those in whom the disorder is to be prevented.

The term "effective amount" means an amount of an active ingredient which is sufficient enough to significantly and positively modify the symptoms and/or conditions to be treated (e.g., provide a positive clinical response). The effective amount of an active ingredient for use in a pharmaceutical composition will vary with the particular condition being treated, the severity of the condition, the duration of the treatment, the nature of concurrent therapy, the particular active ingredient(s) being employed, the particular pharmaceutically-acceptable excipient(s)/carrier(s) utilized, and like factors within the knowledge and expertise of the attending physician.

The term "patient" refers to any animal (e.g., a mammal), including, but not limited to humans, nonhuman primates, rodents, and the like, which is to be the recipient of a particular treatment.

Typically, the term "patient" refers to a human subject.

In embodiments, there is provided a method of treating disease in a patient comprising administering to the patient an effective amount of a compound of the disclosure, or a pharmaceutically acceptable salt thereof, wherein the disease is selected from isolated steatosis, NASH, liver fibrosis and cirrhosis.

In embodiments, there is provided a method of treating a 17PHSD13 mediated disease in a patient comprising administering to the patient an effective amount of a compound of the disclosure, or a pharmaceutically acceptable salt thereof, such as NASH.

The compounds of the present disclosure may be used in the methods described above as either as single pharmacological agents or in combination with other pharmacological agents or techniques. Such combination therapies may be achieved by way of the simultaneous, sequential or separate dosing of the individual components of the treatment. These combination therapies (and corresponding combination products) employ the compounds of the present disclosure and the other pharmacological agent(s).

In embodiments, there is provided a combination for use in the treatment of liver disease, such as NASH, comprising a compound of the disclosure, or a pharmaceutically acceptable salt thereof, and a sodium-glucose transport protein 2 (SGLT2) inhibitor. In further embodiments, the SGLT2 inhibitor is selected from canagliflozin, dapagliflozin, empagliflozin, ertugliflozin, ipragliflozin, luseogliflozin, and remogliflozin.

In embodiments, there is provided a combination for use in the treatment of liver disease, such as NASH, comprising a compound of the disclosure, or a pharmaceutically acceptable salt thereof, and metformin, or a pharmaceutically acceptable salt thereof.

In embodiments, there is provided a combination for use in the treatment of liver disease, such as NASH, comprising a compound of the disclosure, or a pharmaceutically acceptable salt thereof, and a glucagon-like peptide-1 receptor (GLP1) agonist. In further embodiments, the GLP1 agonist is selected from exenatide, liraglutide, lixisenatide, albiglutide, dulaglutide, and semaglutide.

In embodiments, there is provided a combination for use in the treatment of liver disease, such as NASH, comprising a compound of the disclosure, or a pharmaceutically acceptable salt thereof, and a dipeptidyl peptidase 4 (DPP4) inhibitor agonist. In further embodiments, the DPP4 inhibitor is selected sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, anagliptin, teneligliptin, alogliptin, trelagliptin, omarigliptin, evogliptin, gosogliptin, and dutogliptin.

In embodiments, there is provided a combination for use in the treatment of liver disease, such as NASH, comprising a compound of the disclosure, or a pharmaceutically acceptable salt thereof, and a PPAR agonist. In further embodiments, the PPAR agonist is a PPARa agonist. In further embodiments, the PPAR agonist is a PPARy agonist. In further embodiments, the PPAR agonist is a PPARa/y agonist. In further embodiments, the PPAR agonist is selected from clofibrate, gemfibrozil, ciprofibrate, bezafibrate, and fenofibrate. In further embodiments, the PPAR agonist is a thiazolidinedione. In further embodiments, the thiazolidinedione is selected from pioglitazone, rosiglitazone, lobeglitazone, and rivoglitazone. In further embodiments, the PPAR agonist stimulates liver expression of FGF21.

Although the compounds of the disclosure are primarily of value as therapeutic agents for use in patients, they are also useful whenever it is required to inhibit 17PHSD13. Thus, they are useful as pharmacological standards for use in the development of new biological tests and in the search for new pharmacological agents.

Examples

The specification will now be illustrated by the following non-limiting Examples in which, generally:

(i) operations were carried out at rt, i.e. in the range 17 to 28°C and where needed under an atmosphere of an inert gas such as N2;

(ii) where reactions refer to being degassed or purged, this can be performed for example by purging the reaction solvent with a constant flow of nitrogen for a suitable period of time (for example 5 to 10 min) or by repeatedly evacuating the vessel and backfill with appropriate inert atmosphere (for example nitrogen (g) or argon (g));

(iii) in general, the course of reactions was followed by thin layer chromatography (TLC) and/or analytical high performance liquid chromatography (HPLC or UPLC) which was usually coupled to a mass spectrometer (LCMS).

(iv) when necessary, organic solutions were dried over anhydrous MgSCU or NajSC , or by using ISOLUTE Phase Separator, and workup procedures were carried out using traditional phase separating techniques. When a drying agent such as e.g. MgSO4 or Na2SO4 is used for drying an organic layer, it is understood that said organic layer is filtered before concentration of said layer.

(v), evaporations were carried out either by rotary evaporation in vacuo or in a GENEVAC HT-4 / EZ-2 or BIOTAGE V10;

(vi) unless otherwise stated, flash column chromatography was performed on straight phase silica, using either MERCK Silica Gel (Art. 9385) or prep-packed cartridges such as BIOTAGE SNAP cartridges (40-63 pm silica, 4-330 g), BIOTAGE SFAR Silica HC D cartridges (20 pm, 10-100 g), INTERCHIM PURIFLASH cartridges (25 pm, 4-120 g), INTERCHIM PURIFLASH cartridges (50 pm, 25-330 g), GRACE GRACE RESOLV Silica Flash Cartridges (4-120 g) or AGELA FLASH Colum Silica-CS cartridges (80- 330g), or on reversed phase silica using AGELA TECHNOLOGIES C-18, spherical cartridges (20-35pm, 100A, 80-330g), manually or automated using a GRACE REVELERIS X2 Flash system or similar system;

(vii) preparative reversed phase HPLC and preparative reversed phase SFC were performed using standard HPLC and SFC instruments, respectively, equipped with either a MS and/or UV triggered fraction collecting instrument, using either isocratic or a gradient of the mobile phase as described in the experimental section. In some instances the compound may be dissolved in a solvent e.g. DMSO and filtered through a syringe filter prior to purification on preparative HPLC.

Relevant fractions were collected, combined and freeze-dried or evaporated to give the purified compound or relevant fractions were collected, combined and concentrated at reduced pressure, extracted with DCM or EtOAc, and the organic phase was dried either over NajSC or by using a phase-separator, and then concentrated at reduced pressure to give the purified compound;

(viii) yields, where present, are not necessarily the maximum attainable, and when necessary, reactions were repeated if a larger amount of the reaction product was required;

(ix) where certain compounds were obtained as an acid-addition salt, for example a monohydrochloride salt or a di-hydrochloride salt, the stoichiometry of the salt was based on the number and nature of the basic groups in the compound, the exact stoichiometry of the salt was generally not determined, for example by means of elemental analysis data;

(x) in general, the structures of the end-products of the disclosure were confirmed by nuclear magnetic resonance (NMR) and/or mass spectral techniques; proton NMR chemical shift values were measured on the delta scale using BRUKER AVANCE III 300, 400, 500 and 600 spectrometers, operating at ¹ H frequencies of 300, 400, 500 and 600 MHz, respectively. The experiments were typically recorded at 25°C. Chemical shifts are given in parts per million with the solvent as internal standard. Protons on heteroatoms such as NH and OH protons are only reported when detected in NMR and can therefore be missing. In certain instances, protons can be masked or partially masked by solvent peaks and will therefore either be missing and not reported or reported as multiplets overlapping with solvent. The following abbreviations have been used (and derivatives thereof, e.g. dd, doublet of doublets, etc.): s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet; br, broad; qn, quintet; p, pentet. In some cases, the structures of the end-products of the disclosure might appear as rotamers in the NMR-spectrum, in which instances only peaks of the major rotamer are reported. In certain instances, the structures of the intermediates and/or the end-products of the disclosure might appear as rotamers in the NMR-spectrum, in which instances peaks of all rotamers are reported, and only the total number of protons are reported. The ratio of major vs minor rotamer is reported if known. Electrospray mass spectral data were obtained using a WATERS ACQUITY UPLC coupled to a Waters single quadrupole mass spectrometer or similar equipment, acquiring both positive and negative ion data, and generally, only ions relating to the parent structure are reported; high resolution electrospray mass spectral data were obtained using a WATERS XEVO qToF mass spectrometer or similar equipment, coupled to a WATERS ACQUITY UPLC, acquiring either positive and negative ion data, and generally, only ions relating to the parent structure are reported; (xi) intermediates were not necessarily fully purified but their structures and purity were assessed by TLC, analytical HPLC/UPLC, and/or NMR analysis and/or mass spectrometry;

(xii) in general Examples and Intermediate compounds are named using CHEMDRAW PROFESSIONAL version 20.1.1.125 from PerkinElmer. CHEMDRAW PROFESSIONAL version 20.1.1.125 generates the names of chemical structures using the Cahn-lngold-Prelog (CIP) rules for stereochemistry and follows IUPAC rules as closely as possible when generating chemical names. Stereoisomers are differentiated from each other by stereodescriptors cited in names and assigned in accordance with the CIP rules.;

(xiii) in addition to the ones mentioned above, the following abbreviations have been used:

Brine saturated aqueous sodium chloride solution

Calcd calculated

CDI di(lH-lmidazol-l-yl)methanone

DBU 2,3,4,6,7,8,9,10-octahydropyrimido[l,2-o]azepine

DCM dichloromethane

DI PEA /V-ethyl-/V-isopropyl-propan-2-amine

DMF /V,/V-dimethylformamide

DMSO dimethyl sulfoxide

EDC 3-(((ethylimino)methylene)amino)-/V,/\/-dimethylpropan-l-ami ne

ESI electrospray ionization

EtOAc ethyl acetate

EtOH ethanol

FA formic acid

(g) gas

HPLC high performance liquid chromatography

HOBt l-hydroxybenzotriazole;hydrate

HRMS high resolution mass spectrometry

ID inner diameter

LC liquid chromatography

MeCN acetonitrile

MeOH methanol min minute(s)

MS mass spectrometry m/z mass spectrometry peak(s) NCS /V-chlorosuccinimide

NMR nuclear magnetic resonance

OAc acetoxy

PE petroleum ether

Pd-C palladium on charcoal rt room temperature sat saturated

SFC supercritical fluid chromatography

TEA triethylamine

TLC thin layer chromatography

UPLC ultra performance liquid chromatography

UV ultraviolet

Intermediate 1: 5-Fluoro-/V-methyladamantan-2-amine

A solution of 5-fluoroadamantan-2-one (300 mg, 1.8 mmol) and CH3NH2 (30% in EtOH, 5 mL) was stirred at rt overnight. NaBFU (202 mg, 5.4 mmol) was added and the reaction mixture was stirred at rt for another 1 h. The reaction mixture was diluted with water and extracted with EtOAc (3x50 mL). The combined organic layer was washed with brine, dried over anhydrous NajSC and evaporated under vacuum to obtain the title compound (200 mg, crude) as colorless oil; MS m/z [M+H] ⁺ 184

Intermediate 2: tert-Butyl 2-((5-fluoroadamantan-2-yl)(methyl)carbamoyl)hydrazine-l-car boxylate

A solution of tert-butoxycarbohydrazide (343 mg, 2.6 mmol) and CDI (420 mg, 2.6 mmol) in DCM (2 mL) was stirred for 1 h. TEA (315 mg, 3.1 mmol) and 5-fluoro-/V-methyladamantan-2-amine Intermediate 1 (190 mg, 1.0 mmol) were added and stirred at rt for 1 h. The reaction mixture was diluted with MeOH and concentrated under vacuum. The remaining material was purified by reversed phase column chromatography, (gradient 0-100% MeCN in water) to obtain the title compound (160 mg, 45%) as colorless oil; MS m/z [M+H] ⁺ 342 Intermediate 3: /V-(5-Fluoroadamantan-2-yl)-N-methylhydrazinecarboxamide tert-Butyl 2-((5-fluoroadamantan-2-yl)(methyl)carbamoyl)hydrazine-l-car boxylate Intermediate 2 (150 mg, 0.44 mmol) was treated with HCI in 1,4-dioxane (4 M, 2 mL) and stirred at rt for 1 h. The reaction mixture was evaporated under vacuum to afford the hydrochloride of the title compound (140 mg) as white solid; MS m/z [M+H] ⁺ 242.

Intermediate 4: (E)-2-(4-(Difluoromethoxy)-3-hydroxybenzylidene)-/V-(5-fluor oadamantan-2-yl)-/V- methylhydrazine-l-carboxamide

To a solution of /V-(5-fluoroadamantan-2-yl)-/V-methylhydrazinecarboxamide HCI Intermediate 3 (130 mg, 0.5 mmol) in MeOH (1.5 mL) were added NaOAc (53 mg, 0.7 mmol) followed by 4- (difluoromethoxy)-3-hydroxybenzaldehyde (101 mg, 0.5 mmol) and the reaction mixture was stirred at rt for 1 h. The reaction mixture was evaporated under vacuum and the remaining material was purified by reversed phase column chromatography, (gradient: 0-100% MeCN in water containing 10% NH4HCO3) to obtain the title compound (50 mg, 23%) as a brown oil; MS m/z [M+H] ⁺ 412.

Intermediate 5. l-(l-Cyclohexyl-lH-tetrazol-5-yl)-/V-(4-methoxybenzyl)-/V-me thylmethanamine

A mixture of paraformaldehyde (0.550 g, 18.32 mmol) and l-(4-methoxyphenyl)-/V- methylmethanamine (2.77 g, 18.32 mmol) in EtOH (40 mL) was stirred at rt until completion. Isocyanocyclohexane (2 g, 18.32 mmol) was added dropwise to the mixture at 0°C followed by azidotrimethylsilane (2.409 mL, 18.32 mmol). The resulting mixture was stirred at 60°C for 16 h. The solvent was removed under reduced pressure and the residue was purified by straight phase flash chromatography on silica, (gradient: 0-40% EtOAc in PE) to afford the title compound (4.30 g, 74%) as a colourless gum; MS m/z (ESI) [M+H] ⁺ 316.

Intermediate 6: l-(l-Cyclohexyl-lH-tetrazol-5-yl)-/V-methylmethanamine

Pd-C (0.145 g, 1.36 mmol) was added to l-(l-cyclohexyl-lH-tetrazol-5-yl)-/V-(4-methoxybenzyl)-N- methylmethanamine Intermediate 5 (4.3 g, 13.63 mmol) in MeOH (50 mL) at 25°C under E g). The resulting mixture was stirred at 25°C for 16 h. The mixture was filtered through a pad of CELITE and the filtrate was concentrated under reduced pressure to give the title compound (2.50 g, 94%) as a yellow oil; MS m/z (ESI) [M+H] ⁺ 196.

Intermediate 7: /V-((l-Cyclohexyl-lH-tetrazol-5-yl)methyl)-/V-methylcyanamid e l-(l-Cyclohexyl-lH-tetrazol-5-yl)-/V-methylmethanamine Intermediate 6 (200 mg, 0.61 mmol) was added to NCS (107 mg, 0.80 mmol) and Zn(CN)2 (79 mg, 0.68 mmol) in MeCN (6 mL) and water (0.67 mL) at 25°C. The resulting mixture was stirred at 25°C for 16 h. The reaction mixture was diluted with DCM (50 mL), and washed sequentially with water (50 mL) and sat brine (50 mL). The organic layer was dried over NajSC , filtered and evaporated to afford the title compound (50 mg, 37%) as a colorless oil; MS m/z (ESI) [M+H] ⁺ 221.

Intermediate 8: (E)-l-((l-Cyclohexyl-lH-tetrazol-5-yl)methyl)-2-hydroxy-l-me thylguanidine

TEA (2.53 mL, 18.16 mmol) was added to a solution of /V-((l-cyclohexyl-lH-tetrazol-5-yl)methyl)-/V- methylcyanamide Intermediate 7 (2 g, 9 mmol) and hydroxylamine HCI (0.694 g, 9.99 mmol) in MeOH (50 mL) at 25°C under a Nz(g) atmosphere. The resulting mixture was stirred at 60°C for 16 h. The solvent was removed under reduced pressure and the residue was purified by reversed phase flash chromatography on a C18 column (gradient: 0-20% MeCN in water containing 0.1% FA) to afford the title compound (0.650 g, 28%) as a white solid; MS m/z (ESI) [M+H] ⁺ 254.

Example 1: 2-(Difluoromethoxy)-5-(5-((5-fluoroadamantan-2-yl)(methyl)am ino)-l,3,4-oxadiazol-2- yl)phenol h (56 mg, 0.2 mmol) followed by DBU (33 mg, 0.2 mmol) was added to a solution of (Ej-2-(4- (difluoromethoxy)-3-hydroxybenzylidene)-/V-(5-fluoroadamanta n-2-yl)-/\/-methylhydrazine-l- carboxamide Intermediate 3 (45 mg, 0.1 mmol) in MeOH (1 mL) and the reaction mixture was stirred at rt for 1 h. The reaction mixture was purified by preparative HPLC on a XBRIDGE SHIELD RP18 OBD column (5 pm, 150x30 mm ID) using a gradient of 37-60% of MeCN in H2O (10 mM NH4HCO3) as mobile phase to obtain the title compound (16 mg, 36%) as an off-white solid; HRMS (ESI) m/z [M+H] ⁺ calcd for C20H23F3N3O3: 410.1686, found: 410.1690; ^X H NMR (400 MHz, DMSO-dg) 6 1.50 (1H, d), 1.74 (2H, s), 1.78 - 1.97 (5H, m), 2.00 (2H, s), 2.22 (1H, s), 2.66 -2.86 (2H, m), 3.16 (3H, d), 3.54 - 3.86 (1H, m), 6.96 - 7.15 (1H, m), 7.27 (1H, s), 7.30 - 7.38 (1H, m), 7.47 (1H, d), 10.40 (1H, d).

Example 2: 5-(3-(((l-Cyclohexyl-lH-tetrazol-5-yl)methyl)(methyl)amino)- l,2,4-oxadiazol-5-yl)-2- fluorophenol

DIPEA (373 mg, 2.9 mmol), EDC (221 mg, 1.2 mmol) and HOBt (156 mg, 1.2 mmol) followed by /V-[(l- cyclohexyl-l,2,3,4-tetrazol-5-yl)methyl]-/V"-hydroxy-/\/-met hylguanidine Intermediate 8 (292 mg, 1.2 mmol) were added to a solution of 4-fluoro-3-hydroxybenzoic acid (150 mg, 1.0 mmol) in DMF (4 mL). The reaction mixture was stirred at rt overnight and then heated at 60°C for 4 h. The reaction mixture was purified by preparative HPLC on a XBridge Shield RP18 OBD column (30x150 mm ID, 5 pm) using a gradient of 35-55% of MeCN in H2O (10 mM NH4HCO3) to obtain the title compound (78 mg, 22%) as a white solid; HRMS (ESI) m/z [M+H] ⁺ calcd for C17H21FN7O2: 374.1741, found: 374.1739; ^X H NMR (400 MHz, DMSO-d ₆ ) 6: 1.17-1.43 (3H, m), 1.65 (1H, d), 1.70-1.90 (4H, m), 1.94-2.03 (2H, m), 3.06 (3H, s), 4.53-4.64 (1H, m), 5.02 (2H, s), 7.35-7.45(lH, m), 7.46-7.55 (1H, m), 7.58-7.65 (1H, m), 10.58 (1H, s).

In vitro 17bHSD13 enzyme assay

10 concentration of compounds (0.2 pl) in DMSO was added to GREINER PP 384 well plate (781280) using ECHO dispensing (BECKMAN COULTER) followed by 20 pl of recombinant 17bHSD13 (N2-K300). The enzyme reaction was initiated by addition, using CERTUS-FLEX dispenser (GYGER), of 20 pl of substrate solution containing NAD (SIGMA, N1511) and Estradiol (SIGMA, E8875). After each addition plates were centrifuged for 1 min at 150x g (EPPENDORF, 5810R, A-4-81). Final assay conditions were 80 nM of 17bHSD13, 0.5 mM of NAD, 20 pM Estradiol and various concentrations of compound in buffer (5 mM EDTA (TEKNOVA E0306), 0.01% DDM (AFFYMETRIX D310) in 50mM Tris- Cl, pH 7.4). After 2.5 h the reaction were stopped by addition of 20 pl of 0.6 % Formic acid (MERCK 5.33002) and samples were analyzed using LC-MS/MS.

SCIEX LC-MS/MS system: Sample was injected with CTC analytical injector, SHIMATZU LC pumps LC20 and analyzed on the SCIEX API 5000 LCMSMS system with the following settings. Samples were chromatographed on a WATERS, SYMMETRY, C8, 3.5 pm, 2. lx 50 mm) column at constant flow rate of 0.5 mL/min. The mobile phases consist of A (water with 0.2% formic acid) and B (acetonitrile with 0.2% formic acid). The LC gradient profile is as follows: 50% B during 0 to 0.5 min, a linear increase to 100% B during 0.5 to 1 min, hold at 100% B during 1 to 1.6 min then back to 50% B from 1.6 to 2 min. The run time was 2 min with retention times of approximately 0.8 and 1.07 min for Estradiol and Estrone, respectively. Detection was performed on a API 5000 LC/MS/MS system with a triple quadrupole mass spectrometer, a TURBO V ion source, in multiple reaction monitoring (MRM) mode at positive polarity with APCI probe. The MRM pairs were m/z 273.1 to m/z 107.0 and m/z 271.3 to 107.0. for Estradiol and Estrone, respectively. The dwell times were 100 ms for each transition and a depolarization and collision energy of 100 and 40, respectively. Data from MS signals was using area under curve (AUC). Ratio = Estrone/(Estrone + Estradiol)

In vitro 17bHSD4 enzyme assay

10 concentration of compounds (0.2 pl) in DMSO was added to GREINER FLUOTRAC 200 384 well plate (781076) using ECHO dispensing (BECKMAN COULTER). 80 nl of 10 mM Estradiol (SIGMA, E8875) was added using Echo dispensing. The enzyme reaction was initiated by addition, using MULTIDROP COMBI dispensing (THERMO FISHER), of 40 pl of a mix containing recombinant 17bHSD4 (M1-N311) and NAD. Final assay conditions were 40 nM of 17bHSD4, 0.125 mM of NAD, 15 pM Estradiol and various concentrations of compound in buffer (5 M EDTA (TEKNOVA E0306), 0.01% DDM (AFFYMETRIX D310) in 50mM Tris-CI, pH 7.4). After each addition plates were centrifuged for 1 min at 150x g (EPPENDORF, 5810R, A-4-81). NADH formation was measured by fluorescence intensity (Fl) (Ex360/Em460) at time zero (to) and at 1.5 h (ti) in a PHERASTAR FSX (BMG LABTECH). Fl for each sample was calculated as Fl at ti minus Fl at to.

Data analysis

GENEDATA SCREENER was used for curve fitting and calculation of IC ₅ o values.

Compound effect was calculated with the formula below;

Compound % effect = -100 x ((X-min)/(max-min)) where X represents the effect in the presence of test compound, min is DMSO and max is the maximum inhibition of enzyme using a known inhibitor as control.

Table 1

The above description of illustrative embodiments is intended only to acquaint others skilled in the art with the Applicant's specification, its principles, and its practical application so that others skilled in the art may readily adapt and apply the specification in its numerous forms, as they may be best suited to the requirements of a particular use. This description and its specific examples, while indicating embodiments of this specification, are intended for purposes of illustration only. This specification, therefore, is not limited to the illustrative embodiments described in this specification, and may be variously modified. In addition, it is to be appreciated that various features of the specification that are, for clarity reasons, described in the context of separate embodiments, also may be combined to form a single embodiment. Conversely, various features of the specification that are, for brevity reasons, described in the context of a single embodiment, also may be combined to form sub-combinations thereof.