3-Azasteroid compounds for treating mitochondrial function-related diseases

Abstract

Formula (I): 【Chemical 1】 TIFF2025518741000243.tif26170(wherein R 1 , R 2 and R 3 are as defined herein) can be used for the treatment and prevention of neurodegenerative diseases, as well as acute radiation syndrome and myalgic encephalomyelitis (ME, chronic fatigue syndrome) or post-viral syndrome (including chronic symptoms (long COVID) resulting from SARS-CoV2 infection) because the compound can rescue dysfunctional mitochondria.

Description

Technical Field

[0001] The present invention relates to novel compounds for use in the treatment of neurodegenerative diseases, and other conditions involving mitochondrial dysfunction and / or other conditions where it is useful to modulate mitochondrial function. In particular, the present invention relates to bile acid derivatives, pharmaceutical compositions containing them, processes for preparing them, and the use of such compounds in the treatment or prevention of neurodegenerative diseases.

Background Art

[0002] (Background of the Invention) Neurodegenerative diseases are a group of diseases of the central nervous system, including Parkinson's disease, mild cognitive impairment, dementia (including Alzheimer's disease, vascular dementia, and Lewy body dementia), Huntington's disease, and amyotrophic lateral sclerosis (motor neuron disease). Since the incidence of neurodegenerative diseases increases with age, such conditions are becoming an increasingly large problem in societies where the average age of the population is rising. Currently, there is no cure for any of these diseases, although there are some available therapeutic agents that alleviate the symptoms of Parkinson's disease, certain cognitive impairments, and dementia.

[0003] The symptoms of Parkinson's disease are resting tremor, bradykinesia, and rigidity, and these symptoms are caused by neurodegeneration and the loss of dopaminergic neurons. There is a great deal of evidence suggesting a strong association between mitochondrial dysfunction and Parkinson's disease. Mild deficiencies in mitochondrial electron transport system NADH dehydrogenase (complex I) activity are seen in the tissues of Parkinson's disease patients, and several proteins associated with familial Parkinson's disease are either mitochondrial proteins or are associated with mitochondria.

[0004] Alzheimer's disease causes progressive cognitive impairment and is characterized by the presence of extracellular senile plaques and intracellular neurofibrillary changes. Mitochondrial dysfunction is thought to lead to the deposition of β-amyloid protein, the main component of senile plaques, and the formation of neurofibrillary changes.

[0005] Huntington's disease is a hereditary progressive neurodegenerative disease, characterized by movement disorders, personality changes, and cognitive decline. The pathology of Huntington's disease provides evidence of an association with mitochondrial dysfunction.

[0006] Amyotrophic lateral sclerosis is also thought to be associated with mitochondrial dysfunction. This disease targets motor neurons within the central nervous system, resulting in muscle weakness, atrophy, and death within 2 - 3 years of diagnosis.

[0007] Attempts have been made to discover compounds capable of treating neurodegenerative diseases, and several compounds targeting mitochondria have been developed. For example, bile acids such as UDCA (ursodeoxycholic acid) have been shown to have beneficial effects on mitochondrial dysfunction in tissues from specific patients with Parkinson's disease, particularly tissues from patients with the Parkin mutant form of Parkinson's disease (Mortiboys et al., 2013) and tissues from patients with the G2019S LRRK2 mutant form of Parkinson's disease (Mortiboys et al., 2015). Furthermore, known bile acids such as UDCA have been shown to have beneficial effects on fibroblasts from patients with both sporadic Alzheimer's disease and familial Alzheimer's disease due to PSEN1 mutations (Bell et al., 2018). Additionally, additional studies have shown that UDCA is effective in cells from patients with sporadic Parkinson's disease (Carling et al., 2020).

[0008] WO2014 / 036379, WO2015 / 061421, and WO2016 / 145216 teach that bile acids can be used in the treatment of neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease, Huntington's disease, and amyotrophic lateral sclerosis. WO2015 / 061421 relates to deuterated bile acids, and WO2016 / 145216 relates to fluorinated bile acids, particularly bile acids fluorinated at the 3 - position and / or 7 - position. WO2020 / 128514 relates to 2 - fluorinated bile acids having mitochondrial rescue properties.

[0009] Mitochondrial dysfunction is also thought to play a role in acute radiation syndrome (ARS) because mitochondria are sensitive to oxidative stress. There is also evidence that spaceflight can change mitochondrial function and cause DNA damage (da Silveira et al., 2020). The literature by Mhatre et al. (2022) considered the effects of the environment to which astronauts may be exposed during spaceflight, and found that when mice are exposed to radiation, many effects occur, such as increased lipid peroxidation and increased protein oxidation markers, as well as mitochondrial damage. However, pretreatment of mice with the antioxidant MitoQ is thought to reduce this oxidative stress. Compounds capable of rescuing mitochondria may therefore be used for the treatment and prevention of ARS in the case of nuclear accidents or incidents, or when humans or animals are exposed to radiation during space travel.

[0010] Mitochondrial dysfunction is also associated with conditions such as myalgic encephalomyelitis (ME, chronic fatigue syndrome) and chronic symptoms resulting from SARS-CoV2 infection (long COVID) (Wood et al., 2021).

[0011] Therefore, it would be advantageous to develop further compounds capable of rescuing dysfunctional mitochondria. Summary of the Invention

[0012] (Summary of the Invention) In a first aspect of the present invention, formula (I):

Chemical formula

Chemical formula

Chemical formula

Mode for Carrying Out the Invention

[0013] (Detailed Description of the Invention) As used herein, unless otherwise required by express language or necessary implication, the words "comprises", "comprises", "comprising", or variations thereof are used in an inclusive sense, i.e., used to state the presence of the recited features and not to exclude the presence or addition of further features in various embodiments of the invention.

[0014] All publications, including but not limited to patents and patent applications, cited herein are hereby incorporated by reference as if each individual publication were specifically and individually indicated to be incorporated by reference as fully described herein.

[0015] As used herein, reference to "pharmaceutical use" refers to use for administration to a human or animal, particularly a human or mammal, such as a domesticated mammal or an edible domesticated mammal, for the treatment or prevention of a disease or disorder. The term "pharmaceutical composition" refers to a composition suitable for pharmaceutical use, and "pharmaceutically acceptable" refers to an agent suitable for use in a pharmaceutical composition. Other similar terms should be construed accordingly.

[0016] In the present application, the term "C 1-8 " alkyl refers to a straight-chain or branched, fully saturated hydrocarbon group having 1 to 8 carbon atoms. This term includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, s-butyl, and t-butyl. Other alkyl groups, such as C 1-6 alkyl, C1-4 Alkyl, C 1-3 Alkyl, or C 1-2 The alkyl is as defined above, but contains a different number of carbon atoms.

[0017] The term "alkylene" refers to a straight-chain or branched, fully saturated hydrocarbon chain. Preferably, alkylene is C 1-6 Alkylene, C 1-5 Alkylene, C 1-4 Alkylene, C 1-3 Alkylene, or C 1-2 It is alkylene. Examples of alkylene groups include -CH2-, -CH2CH2-, -CH(CH3)-CH2-, -CH2CH(CH3)-, -CH2CH2CH2-, -CH2CH(CH2CH3)-, and -CH2CH(CH2CH3)CH2-.

[0018] The term "C 2-6 Alkenyl" refers to a straight-chain or branched hydrocarbon group having 2 to 6 carbon atoms and containing at least one carbon-carbon double bond. This term encompasses linear alkenyl groups such as CH=CH2, CH2CH=CH2, CH=CHCH3, CH2CH2CH=CH2, CH=CHCH2CH3, CH2CH=CHCH3, CH2CH2CH2CH=CH2, CH=CHCH2CH2CH3, CH2CH=CHCH2CH3, CH2CH2CH=CHCH3, CH=CHCH=CHCH3, and CH2CH=CHCH=CH2, as well as branched alkenyl groups such as CH(CH3)CH=CH2 and CH=C(CH3)CH3. Other alkenyl groups, such as C 2-4 Alkenyl, C 2-3 Alkenyl, and C 3-4 The alkenyl is as defined above, but contains a different number of carbon atoms.

[0019] The term "C 2-6"Alkynyl" refers to a linear or branched hydrocarbon group having 2 to 6 carbon atoms and containing at least one carbon-carbon triple bond. This term includes linear alkynyl groups such as C≡CH, CH2CH≡CH, C≡CCH3, CH2CH2C≡CH, C≡CCH2CH3, CH2C≡CCH2CH3, and CH2C≡CCH=CH2, as well as branched alkynyl groups such as CH(CH3)-C≡CH. Other alkynyl groups, for example C 2-4 alkynyl, C 2-3 alkynyl, and C 3-4 alkynyl is as defined above, but contains a different number of carbon atoms.

[0020] The term "halogen" refers to fluorine, chlorine, bromine, or iodine, and the term "halo" refers to a fluoro group, chloro group, bromo group, or iodo group.

[0021] The term "C 1-6 "Haloalkyl" refers to a linear or branched alkyl group as defined above having 1 to 6 carbon atoms and substituted with one or more halo atoms up to perhalo substitution. Examples include trifluoromethyl, chloroethyl, and 1,1-difluoroethyl. Other haloalkyl groups, for example C 1-5 haloalkyl, C 1-4 haloalkyl, C 1-3 haloalkyl, or C 1-2 haloalkyl is as defined above, but contains a different number of carbon atoms.

[0022] The terms "aryl" and "aromatic" refer to an aromatic cyclic group having 6 to 14 ring carbon atoms (for example, 6 to 10 ring carbon atoms unless otherwise specified) and containing up to 3 rings. When an aryl group contains more than 1 ring, not all rings need to be aromatic. Examples include phenyl, naphthyl, and anthracenyl, as well as partially saturated systems such as tetrahydronaphthyl (for example, 1,2,3,4-tetrahydronaphthyl), indanyl, and indenyl.

[0023] The terms "heteroaryl" and "heteroaromatic" refer to an aromatic cyclic group having 5 to 14 ring atoms (5 to 10 ring atoms, for example, unless otherwise specified), containing at least one heteroatom selected from N, O, and S, and containing up to 3 rings. When a heteroaryl group contains more than 1 ring, not all rings need to be aromatic. Examples include pyridine, pyrimidine, pyrrole, thiophene, furan, thiazole, oxazole, fused systems such as indole, benzimidazole, and benzothiophene, and partially saturated systems such as indoline, isoindoline, and dihydrobenzofuran.

[0024] The terms "carbocyclic" and "carboscyclic" refer to a non-aromatic hydrocarbon ring system having 3 to 10 ring carbon atoms (unless otherwise specified), containing up to 3 rings, which may be fused or joined by spiro bonds, or may be a bridged ring system. A carbocyclic group optionally contains one or more carbon-carbon double bonds. Examples include cycloalkyl groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl; and cycloalkenyl groups such as cyclohexenyl and cycloheptenyl; and groups having a bridged structure such as adamantyl. More preferably, a carboscyclic group is a monocyclic fully saturated (cycloalkyl) ring.

[0025] The terms "heterocyclic" and "heterocyclyl" refer to a non-aromatic ring system having 3 to 10 ring carbon atoms (unless otherwise specified) and at least one heteroatom selected from N, O, and S, containing up to 3 rings, which may be fused or joined by spiro bonds, or may be a bridged ring system. A heterocyclic group may be fully saturated or may contain one or more carbon-carbon or carbon-nitrogen double bonds. Examples include piperidinyl, morpholinyl, thiomorpholinyl, thiazolidinyl, tetrahydrothiophenyl, and tetrahydrothiopyranyl. More preferably, a heterocyclyl group is a monocyclic fully saturated ring.

[0026] The term "oxo" refers to a carbonyl substituent (=O), especially one on a carbocyclic or heterocyclic ring. A ring carbon atom may be substituted with one oxo group, and a ring sulfur atom may be substituted with one or two oxo groups.

[0027] The term "protected NH2 group" refers to an amine protected by any known protecting group. Examples of protected NH2 groups include carbamates such as benzyl carbamate (carboxybenzyl, NHCBz), t-butyl carbamate (NHBoc), and 9-fluorenylmethyl carbamate (NHFmoc). Other suitable NH2 protecting groups include triphenylmethyl (trityl), acetyl, benzyl, and paramethoxybenzyl. Other NH2 protecting groups are well known to those skilled in the art (see, for example, the literature of Wuts, PGM and Greene, TW (2006) "Greene's Protective Groups in Organic Synthesis", 4th Edition, John Wiley & Sons, Inc. Hoboken, NJ, USA).

[0028] The term "protected OH group" refers to a hydroxyl protected by any known protecting group. Examples of this type of protected OH group include R 18 C(O)O (wherein R 18 is C 1-6 alkyl or benzyl, especially methyl). Similarly, silyl ether protecting groups may be used, and OH may likewise be protected as an ether, such as C 1-6 alkyl, benzyl, or p-methoxybenzyl ether. Other suitable OH protecting groups are well known to those skilled in the art (see, for example, the literature of Wuts, PGM and Greene, TW (2006) "Greene's Protective Groups in Organic Synthesis", 4th Edition, John Wiley & Sons, Inc. Hoboken, NJ, USA).

[0029] The salts of the compounds of formula (I) are R 1It may also be an acid addition salt of a quaternary amine formed when the nitrogen atom bonded thereto is quaternized. Alternatively, R 3 When containing C(O)OH or S(O)2OH, the salt may be a base addition salt. R 1 When R is a substituent N(R 11a )(R 11b ) or N(R 14a )(R 14b ), or when R 3 is a substituent N(R 12a )(R 12b ), N(R 13a )(R 13b ), N(R 15a )(R 15b ), or N(R 19a )(R 19b ), or when R 4 contains an amine group, the salt may be formed by quaternization of the amine.

[0030] Any salt intended for administration to a patient would likely be pharmaceutically acceptable, but other salts may also be used during the synthesis of pharmaceutically acceptable end products. Pharmaceutically acceptable salts are known to those skilled in the art and are summarized in the literature by Gupta et al., Molecules, 23, 1719 (2018).

[0031] Pharmaceutically acceptable acid addition salts include hydrochloride, trifluoroacetate, mesylate, hydrobromide, sulfate, and fumarate.

[0032] Pharmaceutically acceptable base addition salts include salts of sodium, potassium, calcium, aluminum, zinc, magnesium, and other metals, as well as choline, amine salts (including triethylamine, N,N - diisopropylethylamine (DIPEA), diethanolamine, ethanolamine, ethylenediamine, meglumine), and other well - known base addition salts.

[0033] The compounds of formula (I) include all stereoisomers. In the compounds of the present invention, the stereochemistry of the bile acid ring system is fixed, and thus the term "stereoisomer" as used herein refers only to the stereoisomers of the R 1 substituent and / or the R 3 substituent in the compound of formula (I), and does not include the stereoisomers of the bile acid ring system.

[0034] The compounds of formula (I) include all isotope variants. The term "isotope variant" refers to an isotope-labeled compound that is identical to that described in formula (I) except that one or more atoms have been replaced by atoms having an atomic mass or mass number different from the most commonly found atomic mass or mass number in nature, or an isotope-labeled compound in which the ratio of atoms having an atomic mass or mass number not commonly found in nature has been increased (the latter concept is referred to as "enriched isotope"). Examples of isotopes that can be incorporated into the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine, iodine, and chlorine, such as 2 H (deuterium), 3 H, 11 C, 13 C, 14 C, 18 F, 123 I, or 125 I (for example, 3 H, 11 C, 14 C, 18 F, 123 I, or 125 I), which may be natural or non-natural isotopes.

[0035] As described above, the present invention provides a compound of formula (I) as defined above, or a salt, solvate, and / or isotope variant thereof.

[0036] In a further aspect of the present invention, formula (IZ):

Chemical formula

Chem.

Chem.

[0037] In some cases, [Chemical formula] is a single bond, and the compound of formula (I) or the compound of formula (IZ) is of formula (IA) or (IB): [Chemical formula] (wherein R 1 , R 3 , and n are as defined above for formula (I) or formula (IZ)).

[0038] Alternatively, [Chemical formula] is a double bond, and the compound of formula (I) or the compound of formula (IZ) is of formula (IC): [Chemical formula] (wherein R 1 , R 3 , and n are as defined above for formula (I) or formula (IZ)).

[0039] In some suitable compounds, the compound of formula (I) or the compound of formula (IZ) is a compound of formula (IA).

[0040] In some suitable compounds, the compound of formula (I) or the compound of formula (IZ) is a compound of formula (IB).

[0041] In some suitable compounds, the compound of formula (I) or the compound of formula (IZ) is a compound of formula (IC).

[0042] In some suitable compounds of the present invention, R 1 is H, C 1-6 alkyl, C 2-6 alkenyl, or C 2-6 alkynyl, more preferably H or C 1-6 alkyl, especially H, methyl, or ethyl. In particularly suitable compounds, R 1 is H.

[0043] In other suitable compounds of the present invention, R 1 is -C(O)R 4 where R 4 is as defined above for formula (I) or formula (IZ).

[0044] In some more suitable compounds of formula (I), R 1 is -C(O)R 4 where R 4 is selected from OH, NH2, NH3 + , and C 1-6 alkyl optionally substituted with one or more substituents selected from 6- to 14-membered aryl or 5- to 14-membered heteroaryl (said aryl and heteroaryl being optionally substituted with one or more substituents selected from OH and halo). In more suitable compounds of this kind, R 4 is selected from OH, NH2, NH3 + , phenyl (optionally substituted with one or more substituents selected from OH and halo), and C 1-6 alkyl optionally substituted with one or more substituents selected from 5- to 10-membered nitrogen-containing heteroaryl groups (such as pyrrole, pyridine, or indole, optionally substituted with one or more substituents selected from OH and halo).

[0045] In particularly suitable compounds of formula (I) where R 1 is -C(O)R 4 where R 1is an amino acid residue or a salt thereof. As used herein, "amino acid residue" refers to an amino acid lacking an OH group, i.e., a substituent of the type -C(O)-C(R)-NH2 (wherein R is an amino acid side chain). The salt of an amino acid residue is -C(O)-C(R)-NH3 + is a substituent of the species. During the synthesis of these compounds, the N-protected amino acid residue can be used as an intermediate. The N-protected amino acid residue is -C(O)-C(R)-NHP 1 (wherein P 1 is an amine protecting group) is a substituent of the species. Suitable amine protecting groups P 1 are as described above, and in particular carbamate-forming groups such as -C(O)O-t-butyl (Boc), -C(O)O-fluorenylmethyl (Fmoc), and -C(O)O-benzyl (CBz).

[0046] In the compounds of formula (I), examples of this type of R 1 groups include residues of glycine, valine, isoleucine, leucine, tryptophan, and tyrosine, and salts of these amino acid residues.

[0047] In some suitable compounds of formula (I), R 1 is -C(O)OR 5 wherein R 5 is as defined above for formula (I) or formula (IZ).

[0048] Further suitable compounds of formula (I) in which R 1 is -C(O)OR 5 wherein R 5 is OH, NH2, NH3 + a C optionally substituted with one or more substituents selected from 6- to 14-membered aryl and 5- to 14-membered heteroaryl (wherein the aryl and heteroaryl are optionally substituted as described above for formula (I) or formula (IZ)) 1-6 alkyl selected from.

[0049] In some even more suitable compounds of formula (I), R 5 is C 1-6It is alkyl (e.g., t-butyl), benzyl, or fluorenylmethyl.

[0050] In particularly suitable compounds of this kind, R 1 is C(O)O-benzyl (CBz).

[0051] Some suitable R 1 is -C(O)R 4 In the compounds of formula (IZ), R 4 is selected from OH, NH2, NH3 + , a protected OH group, a protected NH2 group, and one or more substituents selected from C 1-6 alkyl optionally substituted with one or more substituents selected from OH and halo and optionally substituted with one or more substituents selected from 6- to 14-membered aryl or 5- to 14-membered heteroaryl (the aryl and heteroaryl being optionally substituted with one or more substituents selected from OH and halo). In more suitable compounds of this kind, R 4 is selected from OH, NH2, NH3 + , a protected NH2 group, phenyl optionally substituted with one or more substituents selected from OH and halo, and a 5- to 10-membered nitrogen-containing heteroaryl group (e.g., pyrrole, pyridine, or indole optionally substituted with one or more substituents selected from OH and halo) and optionally substituted with one or more substituents selected from C 1-6 alkyl.

[0052] Particularly suitable, R 1 is -C(O)R 4 In the compounds of formula (IZ), R 1 is an N-protected amino acid residue, an amino acid residue, or a salt thereof. As used herein, "amino acid residue" refers to an amino acid lacking an OH group, i.e., a substituent of the kind -C(O)-C(R)-NH2 (wherein R is an amino acid side chain). A salt of an amino acid residue is a substituent of the kind -C(O)-C(R)-NH3 + , and an N-protected amino acid residue is a substituent of the kind -C(O)-C(R)-NHP 1 (wherein P 1 is an amine protecting group). Suitable amine protecting groups P 1is as described above, and in particular carbamate-forming groups such as -C(O)O-t-butyl (Boc), -C(O)O-fluorenylmethyl (Fmoc), and -C(O)O-benzyl (CBz).

[0053] In the compounds of formula (IZ), this type of R 1 Examples of groups include residues of glycine, valine, isoleucine, leucine, tryptophan, and tyrosine, salts of those amino acid residues, and residues of N-protected glycine, valine, isoleucine, leucine, tryptophan, and tyrosine.

[0054] In other suitable compounds of formula (IZ), R 1 is -C(O)OR 5 wherein, R 5 is OH, NH2, NH3 + , a protected OH group, a protected NH2 group, and C 1-6 alkyl optionally substituted with one or more substituents selected from 6- to 14-membered aryl or 5- to 14-membered heteroaryl (the aryl and heteroaryl are optionally substituted as described above). In more suitable compounds of this type, C(O)OR 5 is an amine protecting group that forms a carbamate together with the nitrogen atom to which it is attached. R 5 is more preferably C 1-6 alkyl optionally substituted with a 6- to 14-membered aryl or 5- to 14-membered aryl group. For example, R 5 may be t-butyl, benzyl, or fluorenylmethyl.

[0055] Some of the compounds of formulas (I), (IZ), (IA), (IB), and (IC) are salts in which the nitrogen atom to which R 1 is attached is quaternized, such as formulas (ID), (IE), (IF), or (IG):

Chemical formula

[0056] In the immediately preceding embodiment, the counterion (anion) Z - is also present. Suitable counterions include chloride salts, trifluoroacetate salts, mesylate salts, bromide salts, sulfate salts, and fumarate salts, especially chloride and trifluoracetate.

[0057] In some suitable compounds, the compound of formula (I) or the compound of formula (IZ) is a compound of formula (ID).

[0058] In some suitable compounds, the compound of formula (I) or the compound of formula (IZ) is a compound of formula (IE).

[0059] In some suitable compounds, the compound of formula (I) or the compound of formula (IZ) is a compound of formula (IF).

[0060] In some suitable compounds, the compound of formula (I) or the compound of formula (IZ) is a compound of formula (IG).

[0061] Suitable counterions for these salts include acid addition salts, especially the above-mentioned acid addition salts that are pharmaceutically acceptable.

[0062] In some compounds of formula (I) and compounds of formula (IZ), n is 1, and in other compounds of formula (I), n is 2.

[0063] As described above, in the compounds of formula (I) and compounds of formula (IZ), R 3 is selected from C(O)OH, C(O)OR 16 , C(O)N(R 6 )-X 1 -R 7 , C(O)N(R 8 )(R 9 ), and C(O)S-R 10 .

[0064] In some suitable compounds of the present invention, R 3 is C(O)OH or C(O)OR 16 wherein R 16 is C 1-8 alkyl or benzyl. In some cases, R 3 is C(O)OH or C(O)(C 1-6 alkyl), for example C(O)OH or C(O)O(C 1-4 alkyl), particularly C(O)OH.

[0065] In other suitable compounds of the present invention, R 3 is C(O)N(R 6 )-X 1 -R 7 wherein R 6 , X 1 and R 7 are as defined above for formula (I) or formula (IZ). Preferably in these compounds, R 6 is H or methyl. In some embodiments, R 6 is H, and in other embodiments R 6 is methyl.

[0066] As described above, X 1 is C 1-6 alkylene optionally substituted as described above for formula (I) or formula (IZ). In some cases, the alkylene group is straight-chain alkylene, and in other cases, the alkylene group is branched-chain alkylene.

[0067] In some cases, X 1 is unsubstituted, and in other cases X 1 is substituted as defined above for formula (I) or formula (IZ). In the compounds of formula (I) and the compounds of formula (IZ), suitable substituents for X 1 include halo, OR 12a , SR 12a , N(R 12a )(R 12b )C(O)OR 12a, phenyl, and 5- or 6-membered heteroaryl (wherein the phenyl and heteroaryl groups are optionally substituted with one or more substituents selected from halo, C 1-6 alkyl, C 1-6 haloalkyl OR 13a , N(R 13a )(R 13b ), NO2, S(O)2OH, and CN); R 12a and R 12b are each independently selected from H and C 1-6 alkyl; and R 13a and R 13b are each independently selected from H, C 1-6 alkyl, and C 1-6 haloalkyl.

[0068] X 1 More suitable substituents for X include halo, OH, O(C 1-4 alkyl), SH, S(C 1-4 alkyl), C(O)OH, C(O)O-(C 1-6 alkyl), NH2, NH(C 1-4 alkyl), N(C 1-4 alkyl)2, and phenyl optionally substituted with one or more substituents selected from OH, halo, O(C 1-3 alkyl), and O(C 1-3 haloalkyl).

[0069] X 1 Some other more suitable substituents for X include halo, OH, O(C 1-4 alkyl), SH, S(C 1-4 alkyl), C(O)OH, C(O)O-(C 1-6 alkyl), and phenyl optionally substituted with one or more substituents selected from OH, halo, O(C 1-3 alkyl), and O(C 1-3 haloalkyl).

[0070] X 1 Other more suitable substituents for X include halo, OH, O(C 1-4 alkyl), S(C 1-4include alkyl), NH2), and phenyl (optionally substituted with halo or OH).

[0071] X 1 More suitable substituents for X include fluoro, OH, methoxy, ethoxy, i - propyloxy, s - butyloxy, t - butyloxy, S - methyl, NH2, C(O)OH, phenyl, and phenyl substituted with OH. X 1 Examples of particularly suitable substituents for X include fluoro, OH, methoxy, ethoxy, i - propyloxy, t - butyloxy, S - methyl, phenyl, and phenyl substituted with OH.

[0072] As described above, R 7 is selected from C(O)OH, C(O)O - (C 1-6 alkyl), S(O)2OH, and S(O)2O - (C 1-6 alkyl). More preferably, R 7 is selected from C(O)OH, C(O)O - (C 1-3 alkyl), and S(O)2OH, particularly C(O)OH and S(O)2OH.

[0073] Particularly suitable groups C(O)N(R 6 ) - X 1 -R 7 include C(O)NH - (CH2)2 - SO2OH (taurine conjugate) and C(O)NH - CH2 - C(O)OH (glycine conjugate). Other suitable amino acid conjugates include O - t - butyl - L - serine (where R 3 is C(O)NH - CH(CH2O - t - Bu) - C(O)OH), β - phenylalanine (where R 3 is C(O)NH - CH(Ph) - CH2 - C(O)OH), serine (where R 3 is C(O)NH - CH(CH2OH) - C(O)OH), 3 - amino - 2 - fluoropropionic acid (where R 3 is C(O)NH - CH2CHF - C(O)OH), methionine (where R 3 is C(O)NH - CH(CH2CH2SMe) - C(O)OH), β - alanine3 is C(O)NH-CH2CH2-C(O)OH), valine (R 3 is C(O)NH-CH(i-Pr)-C(O)OH), isoleucine (R 3 is C(O)NH-CH(CH[Me]CH2Me)-C(O)OH), sarcosine (R 3 is C(O)N(Me)-CH2-C(O)OH), alanine (R 3 is C(O)NH-CH(Me)-C(O)OH), aspartic acid (R 3 is C(O)NH-CH(CH2C(O)OH)-C(O)OH), phenylalanine (R 3 is C(O)NH-CH(CH2Ph)-C(O)OH), 3-aminobutanoic acid (R 3 is C(O)NH-CH(Me)CH2-C(O)OH), leucine (R 3 is C(O)NH-CH( s Bu)-C(O)OH), lysine (R 3 is C(O)NH-(CH2)4-CH(NH2)-C(O)OH), and tyrosine (R 3 is C(O)NH-C(CH2Ph-OH)-C(O)OH) is included.

[0074] Preferably, the amino acid forming this conjugate is in the L configuration.

[0075] In some compounds of formula (I) and compounds of formula (IZ), R 3 is C(O)N(R 8 )(R 9 ), where R 8 and R 9 are as defined above for formula (I) or formula (IZ).

[0076] In some suitable compounds of this kind, R 8 is H, C 1-4Selected from alkyl, cyclopentyl, or cyclohexyl, where the cyclopentyl and cyclohexyl groups are optionally substituted with methyl, OH, methoxy, or fluoro, more preferably being unsubstituted.

[0077] More preferably, R 8 is selected from H, methyl, ethyl, unsubstituted cyclopentyl and unsubstituted cyclohexyl, for example, H, methyl, or unsubstituted cyclohexyl, and particularly is H.

[0078] As described above, when R 3 is C(O)N(R 8 )(R 9 ), R 9 may be H, C 1-6 alkyl, a 3- to 7-membered carbocyclic group, a 3- to 7-membered heterocyclic group, a 6- to 14-membered aryl, or a 5- to 14-membered heteroaryl, and particularly, R 9 is selected from C 1-6 alkyl, a 3- to 7-membered carbocyclic group, a 3- to 7-membered heterocyclic group, a 6- to 14-membered aryl, and a 5- to 14-membered heteroaryl, where the carbocyclic, heterocyclic, aryl, and heteroaryl groups are optionally substituted as defined above for formula (I) or formula (IZ).

[0079] In some suitable compounds of the present invention, R 9 is C 1-6 alkyl, for example, methyl. The alkyl group R 9 may be unsubstituted or substituted as described above. For example, in some compounds of formula (I), R 9 is C 1-6 alkyl, for example, methyl, which may be substituted with a 3- to 7-membered heterocyclic group, particularly a 5- or 6-membered heterocyclic group, such as morpholinyl, piperidinyl, piperazinyl, pyrrolidinyl, or tetrahydrofuryl, where the heterocyclic group may be unsubstituted or substituted as described above for formula (I).

[0080] Some particularly suitable R 3is C(O)N(R 8 )(R 9 ), in the compound of formula (I) or the compound of formula (IZ), R 8 is H, and R 9 is C 1-6 alkyl, such as methyl, which may be unsubstituted or may be substituted as described above for formula (I). For example, R 8 is H, and R 9 is C 1-6 alkyl, such as a 3- to 7-membered heterocyclyl group, especially a 5- or 6-membered heterocyclyl group (such as morpholinyl, piperidinyl, piperazinyl, pyrrolidinyl, or tetrahydrofuryl, more usually morpholinyl, piperidinyl, or piperazinyl, especially morpholinyl)-substituted methyl, and the heterocyclyl group may be unsubstituted or may be substituted as defined above for formula (I).

[0081] In some suitable compounds of formula (I) and compounds of formula (IZ), R 9 is selected from a 3- to 7-membered carbocyclic group, a 3- to 7-membered heterocyclyl group, phenyl, or a 5- or 6-membered heteroaryl (any of which may be unsubstituted or may be substituted as defined above for formula (I) or formula (IZ)).

[0082] R 9 When it is a carbocyclic group, it is more preferably cyclopentyl or cyclohexyl.

[0083] In some particularly suitable R 3 is C(O)N(R 8 )(R 9 ) in the compound of formula (I) or the compound of formula (IZ), R 8 and R 9 are both 3- to 6-membered cycloalkyl rings. For example, R 8 and R 9 are both cyclohexyl.

[0084] In other suitable compounds of formula (I), R 9is a heterocyclyl group. More preferably, R 9 is a 5- or 6-membered heterocyclyl group containing 1 to 3, for example 1 or 2, heteroatoms, especially N and / or O. For example, R 9 is a nitrogen-containing and / or oxygen-containing heterocyclyl group, such as pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, or morpholinyl, especially tetrahydrofuranyl. The heterocycyl group may be unsubstituted or substituted with one or more substituents selected from C 1-4 alkyl, OH, O-(C 1-4 alkyl), C 1-4 haloalkyl, O-(C 1-4 haloalkyl), oxo, phenyl, benzyl, and halo, provided that in this case, the heteroatoms of the heterocyclyl group are not substituted with OH, O-(C 1-4 alkyl), or O-(C 1-4 haloalkyl), and particularly suitable substituents include oxo. The substituents may be on a carbon atom and / or on a heteroatom selected from N and S (when present). In this case, the substituents on the N atom or S atom are not OH, O-(C 1-4 alkyl), or O-(C 1-4 haloalkyl), and the substituents on the N atom are not oxo.

[0085] In a more suitable compound of formula (I), R 3 is C(O)N(R 8 )(R 9 ), R 8 is H, and R 9 is a heterocyclyl group, and particularly more suitable heterocyclyl groups are unsubstituted or substituted as defined above for formula (I).

[0086] R 9 is a 6- to 14-membered aryl group, which may be, for example, phenyl or naphthyl, and especially phenyl. The aryl group R 9may be unsubstituted or, as defined above for formula (I), in particular, halo, C(O)OH, and C(O)O-(C 1-4 alkyl), for example, fluoro, C(O)OH, and C(O)O-(C 1-3 alkyl), and may be substituted with one or more substituents selected therefrom. In some cases, the aryl group R 9 is unsubstituted or, more preferably, is substituted with one substituent. When R 9 is phenyl, the one substituent may be in the 4-position.

[0087] In some more preferred compounds of formula (I) wherein R 3 is C(O)N(R 8 )(R 9 ), R 8 is H and R 9 is phenyl which is optionally substituted as defined above for formula (I).

[0088] When R 9 is a heteroaryl group, it may be a 5- or 6-membered heteroaryl group, particularly a nitrogen-containing heteroaryl group such as pyridyl, for example pyridin-2-yl. The heteroaryl group may be substituted or unsubstituted, and suitable substituents include halo, C(O)OH, and C(O)O-(C 1-4 alkyl), for example, fluoro, C(O)OH, and C(O)O-(C 1-3 alkyl).

[0089] In some more preferred compounds of formula (I) wherein R 3 is C(O)N(R 8 )(R 9 ), R 8 is H and R 9 is a 6- to 14-membered heteroaryl group such as pyridyl, which is unsubstituted or substituted as defined above for formula (I).

[0090] In some compounds of formula (I) and compounds of formula (IZ), R 3 is C(O)N(R 8 )(R 9and R 8 and R 9 together with the nitrogen atom to which they are attached combine to form a 4- to 10-membered heterocyclic group, such as a 5- to 7-membered heterocyclic group or a 5- or 6-membered heterocyclic group, which heterocyclic group optionally contains one or more additional heteroatoms selected from O, N, and S and is optionally substituted with one or more substituents as defined above for formula (I) or formula (IZ).

[0091] In some suitable compounds, the heterocyclic group includes a monocyclic ring, particularly a 4- to 7-membered ring, such as a 5- or 6-membered ring. The ring may contain no additional heteroatoms, and the nitrogen atom to which R 8 and R 9 are attached may be the only heteroatom within the heterocyclic group. Alternatively, the ring may contain one or more additional heteroatoms, for example, one additional heteroatom or two additional heteroatoms. The additional heteroatoms are preferably selected from N, O, and S.

[0092] Examples of monocyclic heterocyclic groups formed by R 8 and R 9 together with the nitrogen atom to which they are attached include morpholine, piperidine, piperazine, pyrrolidine, thiazoline, isothiazoline, thiazolidine, isothiazolidine, oxazoline, isoxazoline, oxazolidine, isoxazolidine, pyrazoline, and pyrazolidine, where the ring is unsubstituted or substituted as defined above. Further suitable examples of monocyclic heterocyclic groups formed by R 8 and R 9 together with the nitrogen atom to which they are attached include piperidine, pyrrolidine, piperazine, morpholine, and isothiazolidine.

[0093] In other suitable compounds, R 8 and R 9And the heterocyclic group formed by them and the nitrogen atoms to which they are attached contains two or more rings, in particular two rings, which may be fused or bridged or joined by a spiro bond. The heterocyclic group contains no additional heteroatoms, and R 8 and R 9 The nitrogen atom to which is attached may be the only heteroatom within the heterocyclic group. Alternatively, the heterocyclic group may contain one or more additional heteroatoms, for example, one additional heteroatom or two additional heteroatoms. The additional heteroatoms are preferably selected from N, O, and S. In some cases, each ring within the system may contain one or two heteroatoms.

[0094] The spiro-bonded group may also contain a 4- or 5-membered ring containing a nitrogen atom, and R 8 and R 9 attached to the nitrogen atom thereof may be joined via a spiro bond to form a 3- to 6-membered ring, in particular a 4- or 5-membered ring, optionally containing a further heteroatom selected from O, N, and S, in particular O. An example of such a system is 2-oxa-6-azospiro{3,3}heptane (i.e., an azetidine ring spiro-bonded to the oxetane ring at the 3-position).

[0095] The fused system may also contain a 5- or 6-membered ring containing a nitrogen atom, and R 8 and R 9 attached to the nitrogen atom thereof may be fused and joined to a 3- to 6-membered ring.

[0096] The bridged system may also contain a 5- or 6-membered ring containing a nitrogen atom, and R 8 and R 9 attached to the nitrogen atom thereof may be joined by a bridge having one atom or two atoms, for example, a bridge selected from -CH2-, -CH2CH2-, -O-, -NH-, and N(C 1-4 alkyl).

[0097] R 8 and R 9When they are combined with the nitrogen atom to which they are attached to form a heterocyclic group, the group may be unsubstituted or substituted. In some compounds, particularly compounds of formula (I), the substituents are C 1-4 alkyl, OH, O-(C 1-4 alkyl), halo, C 1-4 haloalkyl, O-(C 1-4 haloalkyl), C(O)OH, C(O)O(C 1-4 alkyl), benzyl, N(R 15a )(R 15b )(for example, NH2), and oxo, for example C 1-3 alkyl, OH, O-(C 1-3 alkyl), halo, benzyl, NH2, and oxo, selected from. The substituents may be on a carbon atom and / or on a further heteroatom selected from N and S (when present). In this case, the substituents on the N or S atom are not OH, O-(C 1-4 alkyl), or O-(C 1-4 haloalkyl).

[0098] Heterocycles containing no further heteroatoms may have one or more oxo substituents on the carbon atoms, and particularly suitable substituents for the ring carbon atoms include halo, C 1-4 alkyl, C 1-4 haloalkyl, OH, O(C 1-4 alkyl), benzyl, N(R 15a )(R 15b )(for example NH2), and oxo. In particularly suitable compounds of this kind, R 8 and R 9 together with the nitrogen atom to which they are attached may form a piperidone or pyrrolidone ring, for example a 4-piperidone or 3-pyrrolidone ring.

[0099] In heterocyclic groups containing one or more further nitrogen atoms, the ring nitrogen atoms may have substituents, and suitable substituents for the ring nitrogen atoms include C 1-4 alkyl, C 1-4It includes haloalkyl and benzyl. Such a heterocyclic group may not contain additional substituents, or may contain one or more substituents on the ring carbon atoms as described above.

[0100] A heterocyclic group containing one or more sulfur atoms may have one or more substituents, preferably oxo substituents, on the ring sulfur atom. The ring sulfur atom may have one or two oxo substituents. Further, a heterocyclic group containing one or more sulfur atoms may not contain additional substituents, or may contain one or more substituents on the ring carbon atoms as described above. For example, such a ring may have a single oxo substituent on one or more carbon atoms and / or one or two oxo substituents on the sulfur atom.

[0101] In other compounds, particularly compounds of formula (IZ), the substituents are selected from C 1-3 alkyl, OH, O-(C 1-3 alkyl), and halo.

[0102] R 3 is C(O)N(R 8 )(R 9 ) in other suitable compounds, R 8 and R 9 combine with the nitrogen atom to which they are attached to form a 5- to 10-membered heteroaryl group. The heteroaryl group optionally contains one or more additional heteroatoms selected from N, O, and S and is unsubstituted or substituted with one or more substituents selected from halo, NO2, CN, S(O)2OH, C 1-6 alkyl, C 1-6 haloalkyl, O(R 15a ), N(R 15a )(R 15b ), C(O)OH, and C(O)O-(C 1-6 alkyl).

[0103] In some cases, the heteroaryl group is a 5- or 6-membered monocyclic ring, such as a 5-membered monocyclic ring. For example, the heteroaryl group may be pyrrole, imidazole, triazole, or thiazole, particularly pyrrole.

[0104] In other cases, the heteroaryl group may have two rings. In some of the compounds of the present invention, both rings are aromatic, and the nitrogen atom to which R 8 and R 9 are attached is part of a 5-membered ring fused to a further aromatic or heteroaromatic ring. Examples of this type of heteroaryl group include indole and isoindole, especially isoindole. Alternatively, one of the rings may be partially saturated or fully saturated. Preferably in this case, the saturated or partially saturated ring is the ring containing the nitrogen atom to which R 8 and R 9 are attached. The other ring of the heterocyclic group may be a 5- or 6-membered ring such as phenyl, pyridyl, or pyrrolyl.

[0105] In other more suitable compounds of formula (I) or formula (IZ) where R 3 is C(O)N(R 8 )(R 9 ), R 8 and R 9 are combined with the nitrogen atom to which they are attached to form a morpholine, piperidine, or piperazine ring, which ring is unsubstituted or substituted as defined above.

[0106] In the compounds of formula (I) and formula (IZ), particularly suitable groups C(O)N(R 8 )(R 9 ) include: R 8 is H and R 9 is 4-fluorophenyl, R 8 and R 9 are each cyclohexyl, R 8 is H and R 9 is 4-benzoic acid or its C 1-4 alkyl ester, for example isopropyl-4-benzoate, R 8 is H and R 9is tetrahydrofuranyl, especially tetrahydrofuran-3-yl, R 8 and R 9 are combined with the N atom to which they are attached to form an isoindoline ring, R 8 and R 9 are combined with the N atom to which they are attached to form a morpholine ring, R 8 and R 9 are combined with the N atom to which they are attached to form a piperidine each substituted with oxo or a pyrrolidine ring, for example 4-piperidone or 3-pyrrolidone, R 8 and R 9 are combined with the N atom to which they are attached to form a pyrrole ring, are included.

[0107] In yet other compounds of the present invention, R 3 is C(O)S-R 10 where R 10 is as defined above for formula (I) or formula (IZ), but more preferably is OH, halo, or C 1-6 alkyl optionally substituted with phenyl. In even more suitable compounds of this kind, R 10 is C 1-4 alkyl optionally substituted with OH, halo, or phenyl. In particularly suitable compounds, R 10 is benzyl.

[0108] In some suitable compounds of the present invention, R 1 and R 2 are as defined above, and R 3 is C(O)OH or C(O)OR 16 wherein, R 16 is as defined above for formula (I) or formula (IZ).

[0109] In some particularly suitable compounds of the present invention, R 2 is OH, R 3is C(O)OH, and the compound is of formula (IH) or formula (IJ):

Chem.

[0110] In some compounds of formula (IH) and compounds of formula (IJ), n is 1. In other compounds of formula (IH) and compounds of formula (IJ), n is 2.

[0111] In other suitable compounds of the present invention, R 1 is H, R 2 is OH, and n is 1, and the compound is of formula (IK) or formula (IL):

Chem.

[0112] Particularly suitable compounds of the present invention include: tert-Butyl N-(benzyloxycarbonyl)-3-aza-7β-hydroxy-5β-cholan-24-oate (25a); tert-Butyl 3-aza-7β-hydroxy-5β-cholan-24-oate (27a); 3-Aza-7β-hydroxy-5β-cholan-24-oic acid (28a); tert-Butyl N-methyl-3-aza-7β-hydroxy-5β-cholan-24-oate (29a); N-Methyl-3-aza-7β-hydroxy-5β-cholan-24-oic acid (31a and 33); tert-Butyl N-ethyl-3-aza-7β-hydroxy-5β-cholan-24-oate (30); N-Ethyl-3-aza-7β-hydroxy-5β-cholan-24-oic acid (32); tert-Butyl N-glycolyl-3-aza-7β-hydroxy-5β-cholan-24-oate (34a); N-Glycolyl-3-aza-7β-hydroxy-5β-cholan-24-oic acid (40a); N-[(2S)-2-Amino-3-methylbutanoyl]-3-aza-7β-hydroxy-5β-cholan-24-oic acid (41a); N-[(2S,3S)-2-Amino-3-methylpentanoyl]-3-aza-7β-hydroxy-5β-cholan-24-oic acid (42a); N-[(2S)-2-Amino-4-methylpentanoyl]-3-aza-7β-hydroxy-5β-cholan-24-oic acid (43a); N-[(2S)-2-Amino-3-(1H-indol-3-yl)propanoyl]-3-aza-7β-hydroxy-5β-cholan-24-oic acid (44a); N-[(2S)-2-Amino-3-(4-hydroxyphenyl)propanoyl]-3-aza-7β-hydroxy-5β-cholan-24-oic acid (45a); tert-Butyl N-(benzyloxycarbonyl)-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oate (25b); tert-Butyl 3-aza-7β-hydroxy-25-homo-5β-cholan-25-oate (27b); 3-Aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid (28b); tert-Butyl N-methyl-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oate (29b); N-Methyl-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid (31b); tert-Butyl N-glycolyl-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oate (34b); N-Glycolyl-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid (40b); N-[(2S)-2-Amino-3-methylbutanoyl]-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid (41b); N-[(2S,3S)-2-Amino-3-methylpentanoyl]-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid (42b); N-[(2S)-2-Amino-4-methylpentanoyl]-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid (43b, Example 25); N-[(2S)-2-Amino-3-(1H-indol-3-yl)propanoyl]-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid (44b); N-(Benzyloxycarbonyl)-3-aza-7β-hydroxy-5β-cholan-24-oic acid (50a); N-(3-Aza-7β-hydroxy-5β-cholan-24-amide)-ethylsulfonic acid (51a); N-(3-Aza-7β-hydroxy-5β-cholan-24-amide)-acetic acid (52a); N-(3-Methyl-aza-7β-hydroxy-5β-cholan-24-oil)-(2S)-2-amino-3-[(2-methylpropan-2-yl)oxy]propanoic acid (53a); N-(3-Methyl-aza-7β-hydroxy-5β-cholan-24-oil)-(R)-3-amino-3-phenylpropanoic acid (54a); N-(3-Methyl-aza-7β-hydroxy-5β-cholan-24-oil)-1-amino-4-fluorobenzene (55a); N-(3-Aza-7β-hydroxy-5β-cholan-24-oil)-morpholine (56a); N-(3-Aza-7β-hydroxy-5β-cholan-24-oil)-(S)-2-amino-3-hydroxypropanoic acid (57a); N-(3-Aza-7β-hydroxy-5β-cholan-24-oil)-3-amino-2-fluoropropanoic acid (58a); N-(Cyclohexyl)-N-(3-methyl-aza-7β-hydroxy-5β-cholan-24-oil)-cyclohexanamine (59a); N-(3-Aza-7β-hydroxy-5β-cholan-24-oil)-4-aminobenzoic acid (60a); N-(3-Aza-7β-hydroxy-5β-cholan-24-oil)-(S)-2-amino-4-(methylthio)butanoic acid (61a); N-(3-Aza-7β-hydroxy-5β-cholan-24-amide)-propanoic acid (62a); N-(3-Aza-7β-hydroxy-5β-cholan-24-oil)-(isopropyl-4-aminobenzoate) (63a); S-(3-Aza-7β-hydroxy-5β-cholan-24-oil)-phenylmethanethiol (64a); N-(3-Aza-7β-hydroxy-5β-cholan-24-oil)-(S)-2-amino-3-methylbutanoic acid (65a); N-(3-Aza-7β-hydroxy-5β-cholan-24-oil)-(2S,3S)-2-amino-3-methylpentanoic acid (66a); N-Methyl-N-(3-aza-7β-hydroxy-5β-cholan-24-oil)-glycine (67a); N-(3-Aza-7β-hydroxy-5β-cholan-24-oil)-(S)-2-aminopropanoic acid (68a); N-(Benzyloxycarbonyl)-3-aza-7β-hydroxy-24-homo-5β-cholan-25-oic acid (50b); N-(3-Aza-7β-hydroxy-27-homo-5β-cholan-27-oil)-(S)-2-aminobutanedioic acid (70b); N-(3-Aza-7β-hydroxy-27-homo-5β-cholan-27-oil)-(2S)-2-amino-3-[(2-methylpropan-2-yl)oxy]propanoic acid (71b); N-(3-Aza-7β-hydroxy-27-homo-5β-cholan-27-amide)-ethylsulfonic acid (72b); N-(3-Aza-7β-hydroxy-27-homo-5β-cholan-27-oil)-(S)-2-amino-3-phenylpropanoic acid (73b); N-(3-Aza-7β-hydroxy-27-homo-5β-cholan-27-oil)-3-aminobutanoic acid (74b); N-(3-Aza-7β-hydroxy-27-homo-5β-cholan-27-oil)-(S)-2-amino-4-methylpentanoic acid (75b); N-(3-Aza-7β-hydroxy-27-homo-5β-cholan-27-oil)-(2S)-2,6-diaminohexanoic acid (76b); N-(3-Aza-7β-hydroxy-27-homo-5β-cholan-27-oil)-(2S)-2-amino-3-(4-hydroxyphenyl)propanoic acid (77b); N-{Benzyloxycarbonyl)-3-aza-7β-hydroxy-5β-cholan-24-oil}-2-oxa-6-azaspiro{3,3}heptane (78a); N-{Benzyloxycarbonyl)-3-aza-7β-hydroxy-5β-cholan-24-oil}-4-piperidone (79a); N-{Benzyloxycarbonyl)-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oil}-3-aminotetrahydrofuran (80b); N-{3-Aza-7β-hydroxy-5β-cholan-25-oil}-isoindoline (81a); N-{3-Aza-7β-hydroxy-25-homo-5β-cholan-25-oil}-3-aminotetrahydrofuran (82b); and, Its salts and solvates are included.

[0113] Some more suitable compounds of the present invention include: 3-Aza-7β-hydroxy-5β-cholan-24-oic acid (28a); N-Ethyl-3-aza-7β-hydroxy-5β-cholan-24-oic acid (32); 3-Aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid (28b); N-Methyl-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid (31b); N-Glycolyl-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid (40b); N-[(2S)-2-Amino-3-methylbutanoyl]-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid (41b); and, its salts and solvates are included.

[0114] Other more suitable compounds of the present invention include: N-(3-Aza-7β-hydroxy-5β-cholan-24-amide)-acetic acid (52a); N-(3-Methyl-aza-7β-hydroxy-5β-cholan-24-oyl)-(R)-3-amino-3-phenylpropanoic acid (54a); N-(3-Methyl-aza-7β-hydroxy-5β-cholan-24-oyl)-1-amino-4-fluorobenzene (55a); N-(3-Aza-7β-hydroxy-5β-cholan-24-oyl)-(S)-2-amino-3-hydroxypropanoic acid (57a); N-(3-Aza-7β-hydroxy-5β-cholan-24-amide)-propanoic acid (62a); S-(3-Aza-7β-hydroxy-5β-cholan-24-oyl)-phenylmethanethiol (64a); N-(3-Aza-7β-hydroxy-5β-cholan-24-oyl)-(S)-2-amino-3-methylbutanoic acid (65a); N-(3-Aza-7β-hydroxy-5β-cholan-24-oyl)-(2S,3S)-2-amino-3-methylpentanoic acid (66a); N-Methyl-N-(3-aza-7β-hydroxy-5β-cholan-24-oyl)-glycine (67a); N-(3-Aza-7β-hydroxy-27-homo-5β-cholan-27-oil)-(S)-2-amino-4-methylpentanoic acid (75b); N-(3-Aza-7β-hydroxy-27-homo-5β-cholan-27-oil)-(2S)-2-amino-3-(4-hydroxyphenyl)propanoic acid (77b); N-{Benzyloxycarbonyl)-3-aza-7β-hydroxy-5β-cholan-24-oil}-2-oxa-6-azaspiro{3,3}heptane (78a); N-{Benzyloxycarbonyl)-3-aza-7β-hydroxy-5β-cholan-24-oil}-4-piperidone (79a); N-{Benzyloxycarbonyl)-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oil}-3-aminotetrahydrofuran (80b); and, Its salts and solvates are included.

[0115] In some embodiments of the present invention, the compound is N-(Benzyloxycarbonyl)-3-aza-7β-hydroxy-24-homo-5β-cholan-25-oic acid (50b); N-(3-Aza-7β-hydroxy-27-homo-5β-cholan-27-oil)-(S)-2-amino-3-phenylpropanoic acid (73b); Not N-(3-Aza-7β-hydroxy-27-homo-5β-cholan-27-oil)-3-aminobutanoic acid (74b), or None of its salts or solvates.

[0116] (Preparation of the Compounds of the Invention) R 1 is C(O)OR 5 and R 3 is C(O)OR 16 The compound of formula (I) wherein is of formula (II):

Chemical formula

[0117] The compounds of formula (II) are novel and form a further aspect of the present invention.

[0118] The compounds of formula (II) are each of formula (III):

Chemical formula

[0119] Preferably, the reaction is carried out under weakly basic conditions, such as in the presence of an aqueous sodium carbonate solution. The solvent may also include an organic solvent, such as dichloromethane.

[0120] The compounds of formula (III) may be prepared by oxidation of a compound of formula (V):

Chemical formula

[0121] Oxidation may be carried out using an oxidizing agent such as (diacetoxyiodo)benzene. In that case, the reaction is preferably carried out at room temperature, for example, about 15 to 25 °C, in a polar organic solvent such as a mixture of acetonitrile and water.

[0122] The compound of formula (V) may be prepared by reaction of a lactone of formula (VI):

Chemical formula

[0123] The reaction may be carried out at an elevated temperature, for example, about 80 to 100 °C, preferably in a sealed tube.

[0124] The compound of formula (VI) may be prepared by oxidation of a compound of formula (VII):

Chemical formula

[0125] Suitable oxidizing agents include meta-chloroperbenzoic acid (mCPBA), and the reaction is preferably carried out at room temperature, for example, about 15 to 25 °C, in an organic solvent such as dichloromethane.

[0126] The compound of formula (VII) may be prepared by oxidation of a compound of formula (VIII):

Chemical formula

[0127] Suitable oxidants for this reaction include the use of (diacetoxyiodo)benzene in combination with a catalytic amount of 2,2,6,6 - tetramethylpiperidine 1 - oxyl radical (TEMPO) or a TEMPO derivative. The reaction may be carried out in a dry state in an organic solvent, for example, dichloromethane.

[0128] The compound of formula (VIII) where n is 1 is a compound of formula (IX):

Chemical formula

[0129] The compounds of formula (IX) and the compounds of formula (X) are readily available and may also be prepared by methods known to those skilled in the art. Among the compounds of formula (IX),

Chemical formula

Chemical formula

[0130] The compound of formula (VIII) where n is 2 may be prepared from the compound of formula (IX) according to the following reaction scheme.

Chemical formula

[0131] Step b: In this step, the compound of (XXIII) is reacted with a compound of the formula: P 2 -X (wherein P 2 is a protecting group, for example a silyl protecting group such as t-butyldimethylsilyl chloride, etc.) in the presence of a base, for example imidazole. The reaction may be carried out at room temperature. The product is a protected compound of formula (XXII), where R 20 is as defined above for formula (XXIII), P 2 is as defined above, and R 2a is

Chemical formula

Chemical formula

[0132] Step c: In this step, the compound of formula (XXII) is reduced to obtain an alcohol of formula (XXI) (wherein P 2 and R 2a are as defined above for formula (XXII)). Suitable reducing agents include lithium borohydride, and the reaction is preferably carried out at a low temperature, generally about -5 to 5 °C, for example 0 °C, in an organic solvent, for example tetrahydrofuran.

[0133] Step d: The compound of formula (XXI) is oxidized using a suitable oxidizing agent, for example oxalyl chloride, to obtain a compound of formula (XX) (wherein P 2 and R2a To obtain the product as defined above for formula (XXII). The reaction may be carried out at a temperature of about -78 °C.

[0134] Step e: React the compound of formula (XX) with a compound of formula (XIX) (wherein R 3a is as defined for formula (II)). The reaction may be carried out in a polar organic solvent, for example dichloromethane, at a temperature of about 15 - 25 °C, generally at room temperature. The product is a compound of formula (XVII), where R 3a is as defined for formula (II), and P 2 and R 2a are as defined for formula (XXII).

[0135] Step f: Reduce the compound of formula (XVII) to obtain a compound of formula (XVI) (wherein R 3a is as defined for formula (II), and P 2 and R 2a are as defined for formula (XXII)). Preferably, the reduction is carried out by hydrogenation with a suitable catalyst, such as palladium on carbon. Suitable reaction solvents include organic solvents, such as ethyl acetate, and the reaction may be carried out at a temperature of about 15 - 25 °C, generally at room temperature.

[0136] Step g: Deprotect the compound of formula (XVI) to obtain a compound of formula (XV) (wherein R 3a is as defined for formula (II)). The deprotection may be achieved by treatment with an acid, such as hydrochloric acid in water.

[0137] Step h: The compound of formula (XV) may be converted to the compound of formula (VIII) by oxidation. Suitable oxidizing agents for this reaction include the combination of (diacetoxyiodo)benzene and a catalytic amount of TEMPO or a TEMPO derivative. The reaction may be carried out in a dry state in an organic solvent, for example dichloromethane. The reaction may be carried out at a temperature of about 15 - 25 °C, generally at room temperature.

[0138] The compound of formula (I) may be converted to other compounds of formula (I). For example, R 1 wherein the compound of formula (I) where R 1 is H can be prepared by hydrogenation with a palladium catalyst from a compound of formula (I) (wherein R 5 is C(O)OR 5 as defined above for formula (IZ)). Preferably, the hydrogenation is carried out in an alcoholic solvent, for example, methanol.

[0139] In the compound of formula (I), C(O)OR 5 is preferably an amine protecting group, which forms a carbamate together with the nitrogen atom to which it is attached. R 5 is more preferably C 1-6 alkyl optionally substituted with a 6- to 14-membered aryl or 5- to 14-membered heteroaryl group. For example, R 5 may be t-butyl, benzyl, or fluorenylmethyl.

[0140] R 1 is C(O)OR 5 Among the specific compounds of formula (I), there are: tert-butyl N-(benzyloxycarbonyl)-3-aza-7β-hydroxy-5β-cholan-24-oate (25a); tert-butyl N-(benzyloxycarbonyl)-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oate (25b) N-(benzyloxycarbonyl)-3-aza-7β-hydroxy-5β-cholan-24-oic acid (50a); N-(benzyloxycarbonyl)-3-aza-7β-hydroxy-24-homo-5β-cholan-25-oic acid (50b); N-{(benzyloxycarbonyl)-3-aza-7β-hydroxy-5β-cholan-24-oyl}-2-oxa-6-azaspiro{3,3}heptane (78a); N-{(benzyloxycarbonyl)-3-aza-7β-hydroxy-5β-cholan-24-oyl}-4-piperidone (79a); N-{(Benzyloxycarbonyl)-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic}-3-aminotetrahydrofuran (80b); and its salts and solvates are included.

[0141] R 1 is H and R 3 is C(O)OR 16 The compound of formula (I) wherein is converted by reacting with a compound of formula (XX): 1 R 1-6 is C 3 alkyl and R 16 is C(O)OR R 1b -C(O)H (XX) (wherein R 1b is C 1-5 alkyl) and then hydrogenating with a palladium / carbon catalyst.

[0142] This reaction may be carried out in an aqueous solvent.

[0143] R 1 is H and R 3 is C(O)OR 16 The compound of formula (I) wherein is converted by reacting the starting material with a compound of formula (XVIII): 1 R 2-6 is C 2-6 alkenyl or C 3 alkynyl and R 16 is C(O)OR R 1c -X (XVIII) (wherein R 1c is C 2-6 alkenyl or C 2-6 alkynyl and X is a leaving group such as halo, for example chloro or bromo) in the presence of a base.

[0144] R 2The process for a compound of formula (I) where is OH may further include a first step of reacting a compound of formula (I) having a suitable protecting group as a starting material, and a final step of deprotecting the reaction product with a compound of formula (XVIII).

[0145] If necessary, R 1 is H and R 3 is C(O)OR 16 The product where is may be converted to a compound of formula (I) having different R 3 groups by one of the following methods.

[0146] R 3 is C(O)OR 16 The compound of formula (I) where may be hydrolyzed with an acid or a base to obtain a compound of formula (I) where is C(O)OH. The hydrolysis may particularly be carried out using an acid, for example, trifluoroacetic acid. The reaction is preferably carried out at a low temperature, for example, about -5 to 5 °C, in an anhydrous polar organic solvent, for example, dichloromethane. 3 The compound of formula (I) where is H may be converted to a salt of formula (I) where the nitrogen atom to which is attached is quaternized by treatment with an acid, for example, hydrochloric acid or trifluoroacetic acid. This reaction is particularly useful for converting a compound of formula (I) where is H and is C(O)OH or C(O)OR

[0147] R 1 is H 1 to a quaternary ammonium salt of formula (I) where is C(O)OH. 1 is H and R 3 is C(O)OH or C(O)OR 16 where is 3 is C(O)OH

[0148] R 1 is C(O)R 4 and R 3 is C(O)OR 16 The compound of formula (I) where is 1 is H and R 3 is C(O)OR 16 from a compound of formula (I) where is R 4 '-C(O)OH (XXI) (wherein R 4 ' is OR 14a , N(R 14a )(R 14b ), NH3 + , C(O)N(R 14a )(R 14b ), SR 14 a protected OH group, a protected NH2 group, a protected C(O)NH2 group, a 5- or 6-membered nitrogen-containing heterocycle, and a 6- to 14-membered aryl or 5- to 14-membered heteroaryl (the aryl and heteroaryl are optionally substituted with one or more substituents selected from OH, halo, NH2, NO2, S(O)2OH, C 1-6 alkyl, C 1-6 haloalkyl, O(C 1-6 alkyl), and O(C 1-6 haloalkyl)) optionally substituted with one or more substituents selected from the group consisting of) is prepared by reacting with a compound of C 1-6 alkyl under basic conditions, for example using N,N-diisopropylethylamine, and in the presence of a coupling agent, and removing the protecting group if necessary to obtain a group R 4 containing an OH, NH2 or C(O)NH2 group.

[0149] Suitable coupling reagents include known peptide coupling agents such as O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU), O-(benzotriazol-1-yl)-N,N,N',N-tetramethyluronium tetrafluoroborate (TBTU), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate (TATU), (benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate (BOP), (benzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate (PyBOP) carbodiimides such as 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI), and triazoles such as 1-hydroxy-7-azabenzotriazole (HOAt) or hydroxybenzotriazole (HOBt); and chloroformates such as isobutyl chloroformate.

[0150] The product may be converted as described above by reaction with an acid to a compound of formula (I) where R 3 is C(O)OH. In this case, as in the case of treatment with trifluoroacetic acid, removal of the protecting groups from the protected OH and NH2 groups in R 4 will be necessary.

[0151] The product is of formula (IP2):

Chemical formula

[0152] For a protected NH2 group, a protected OH group, or a protected C(O)NH2 group, removal of the protecting group occurs upon acid treatment, and R 2 and n are as defined above and R 3 is C(O)OH and R 1 is C(O)R 4 a compound of formula (I) (wherein R 4 is C alkyl substituted with NH2 or OH or C(O)NH2 1-6 alkyl) an acid addition salt of is obtained. Suitable protected NH2 groups include acid-labile carbamates such as t-butyl carbamate. Suitable protected OH groups include ethers such as monomethyl ether or tetrahydropyran, and suitable protected C(O)NH2 groups include alkyl-substituted amides such as C(O)NH-tBu.

[0153] The compound of formula (IP2) is novel and forms a further aspect of the present invention, formula (IP2):

Chemical formula

[0154] Specific compounds of formula (IP2) include: tert-butyl N-{ (2S)-2-[(tert-butoxycarbonyl)amino]-3-methylbutanoyl}-3-aza-7β-hydroxy-5β-cholan-24-oate (35a); tert-butyl N-{ (2S,3S)-2-[(tert-butoxycarbonyl)amino]-3-methylpentanoyl}-3-aza-7β-hydroxy-5β-cholan-24-oate (36a); tert-Butyl N-{ (2S)-2-[(tert-butoxycarbonyl)amino]-4-methylpentanoyl}-3-aza-7β-hydroxy-5β-cholan-24-oate (37a); tert-Butyl N-{ (2S)-2-[(tert-butoxycarbonyl)amino]-3-[1H-indol-3-yl]propanoyl}-3-aza-7β-hydroxy-5β-cholan-24-oate (38a); tert-Butyl N-{ (2S)-2-[(tert-butoxycarbonyl)amino]-3-[4-hydroxyphenyl]propanoyl}-3-aza-7β-hydroxy-5β-cholan-24-oate (39a); tert-Butyl N-{ (2S)-2-[(tert-butoxycarbonyl)amino]-3-methylbutanoyl}-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oate (35b); tert-Butyl N-{ (2S,3S)-2-[(tert-butoxycarbonyl)amino]-3-methylpentanoyl}-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oate (36b); tert-Butyl N-{ (2S)-2-[(tert-butoxycarbonyl)amino]-4-methylpentanoyl}-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oate (37b); tert-Butyl N-{ (2S)-2-[(tert-butoxycarbonyl)amino]-3-[1H-indol-3-yl]propanoyl}-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oate (38b); and salts and solvates thereof are included.

[0155] R 3 is C(O)N(R 6 )-X 1 -R 7 The compound of formula (I) in which is a compound of formula (I) in which R 3 is C(O)OH, from the formula (XXV): HN(R 6 )-X 1 -R 7(XXV) (wherein R 6 , R 7 and X 1 are as defined for formula (I)) with a coupling reagent in the presence of a base, for example in the presence of an amine such as diisopropylethylamine (DIPEA) or triethylamine (TEA), in an organic solvent such as DMF, may be prepared by reacting. Suitable coupling agents are as described above.

[0156] Similarly, a compound of formula (I) in which R 3 is C(O)N(R 8 )(R 9 ) may be prepared by reacting a compound of formula (I) in which R 3 is C(O)OH with a compound of formula (XXVI): HN(R 8 )(R 9 ) (XXVI); (wherein R 8 and R 9 are as defined for formula (I)) in the presence of a base and in the presence of a coupling agent. Suitable coupling agents are as described above.

[0157] R 3 is C(O)S-R 10 a compound of formula (I) may be prepared by reacting a compound of formula (I) in which R 3 is C(O)OH with a compound of formula (XXVII): HSR 10 (XXVII) (wherein R 10 is as defined for formula (I)).

[0158] Compounds of formula (XXV), compounds of formula (XXVI), and compounds of formula (XXVII) are known and readily available, or may be synthesized by methods known to those skilled in the art.

[0159] (Therapeutic method) Surprisingly, the compounds of the present invention have been shown to be capable of restoring mitochondrial function and of passing through the blood-brain barrier. Accordingly, the present compounds are useful for the treatment or prevention of neurodegenerative diseases including Parkinson's disease, mild cognitive impairment, dementia (including Alzheimer's disease, vascular dementia, dementia with Lewy bodies, and frontotemporal dementia (FTD)), Huntington's disease, amyotrophic lateral sclerosis (motor neuron disease), progressive supranuclear palsy, and Wilson's disease.

[0160] In the following description, references to the compounds of formula (I) and the compounds of formula (IZ) for use in medicine, to the use of the compounds of formula (I) and the compounds of formula (IZ) for the preparation of pharmaceutical compositions, to methods of treatment utilising the compounds of formula (I), and to pharmaceutical compositions comprising the compounds of formula (I) and the compounds of formula (IZ) apply equally to pharmaceutically acceptable salts and solvates of the compounds of formula (I) and the compounds of formula (IZ).

[0161] When the compounds of formula (IZ) are used for medical purposes, R 1 is preferably not C(O)OR 5 and; R 1 is C(O)R 4 in the case of, R 5 is preferably not substituted with a protected OH group, a protected NH2 group, or a protected C(O)NH2 group; and R 3 is C(O)N(R 6 )-X 1 -R 7 in the case of, X 1 is preferably not substituted with a protected OH group, a protected NH2 group, or a protected C(O)NH2 group.

[0162] The compounds of formula (I) and the compounds of formula (IZ) are also useful for the treatment or prevention of conditions in which modulation of mitochondrial function is effective, particularly neurodegenerative diseases such as Parkinson's disease, mild cognitive impairment, dementia (including Alzheimer's disease, vascular dementia, Lewy body dementia, and FTD), Huntington's disease, amyotrophic lateral sclerosis (motor neuron disease), progressive supranuclear palsy, and Wilson's disease.

[0163] The compounds of formula (I) and the compounds of formula (IZ) are also useful for the treatment or prevention of acute radiation syndrome, for example, in the treatment of human patients or diseased animals exposed to or potentially exposed to radiation, such as astronauts during spaceflight.

[0164] The compounds of the present invention are also useful for the treatment or prevention of post-viral syndromes including myalgic encephalomyelitis (ME, chronic fatigue syndrome) and chronic symptoms resulting from SARS-CoV2 infection (long COVID).

[0165] In a further aspect of the present invention, there is provided a compound of formula (I) or a compound of formula (IZ) for use in medicine.

[0166] Similarly provided are: · A compound of formula (I) or a compound of formula (IZ) for use in the treatment of neurodegenerative diseases, · A compound of formula (I) or a compound of formula (IZ) for use in the prevention of neurodegenerative diseases, · A compound of formula (I) or a compound of formula (IZ) for use in the treatment of acute radiation syndrome, · A compound of formula (I) or a compound of formula (IZ) for use in the prevention of acute radiation syndrome, · A compound of formula (I) or a compound of formula (IZ) for use in the treatment of post-viral syndromes including myalgic encephalomyelitis (ME, chronic fatigue syndrome) or chronic symptoms resulting from SARS-CoV2 infection (long COVID), · A compound of formula (I) or a compound of formula (IZ) for use in the prevention of post-viral syndromes including myalgic encephalomyelitis (ME, chronic fatigue syndrome) or chronic symptoms (long COVID) resulting from SARS-CoV2 infection.

[0167] The present invention likewise provides the following: · Use of a compound of formula (I) or a compound of formula (IZ) in the preparation of a therapeutic agent for a neurodegenerative disease, · Use of a compound of formula (I) or a compound of formula (IZ) in the preparation of a prophylactic agent for a neurodegenerative disease, · Use of a compound of formula (I) or a compound of formula (IZ) in the preparation of a therapeutic agent for acute radiation syndrome, · Use of a compound of formula (I) or a compound of formula (IZ) in the preparation of a prophylactic agent for acute radiation syndrome, · Use of a compound of formula (I) or a compound of formula (IZ) in the preparation of a therapeutic agent for post-viral syndromes including myalgic encephalomyelitis (ME, chronic fatigue syndrome) or chronic symptoms (long COVID) resulting from SARS-CoV2 infection, · Use of a compound of formula (I) or a compound of formula (IZ) in the preparation of a prophylactic agent for post-viral syndromes including myalgic encephalomyelitis (ME, chronic fatigue syndrome) or chronic symptoms (long COVID) resulting from SARS-CoV2 infection.

[0168] The present invention further provides the following: · A method of treating a neurodegenerative disease, said method comprising administering to a patient in need thereof an effective amount of a compound of formula (I) or a compound of formula (IZ), · A method of preventing a neurodegenerative disease, said method comprising administering to a patient in need thereof an effective amount of a compound of formula (I) or a compound of formula (IZ), · A method of treating acute radiation syndrome, said method comprising administering to a patient in need thereof an effective amount of a compound of formula (I) or a compound of formula (IZ), · A method of preventing acute radiation syndrome, said method comprising administering to a patient in need thereof an effective amount of a compound of formula (I) or a compound of formula (IZ), · A method for treating a post-viral syndrome including myalgic encephalomyelitis (ME, chronic fatigue syndrome) or chronic symptoms (long COVID) resulting from SARS-CoV2 infection, said method comprising administering to a patient in need thereof an effective amount of a compound of formula (I) or a compound of formula (IZ). · A method for preventing a post-viral syndrome including myalgic encephalomyelitis (ME, chronic fatigue syndrome) or chronic symptoms (long COVID) resulting from SARS-CoV2 infection, said method comprising administering to a patient in need thereof an effective amount of a compound of formula (I) or a compound of formula (IZ).

[0169] Examples of neurodegenerative diseases include Parkinson's disease, mild cognitive impairment, dementia (including Alzheimer's disease, vascular dementia, Lewy body dementia, and FTD), Huntington's disease, amyotrophic lateral sclerosis (motor neuron disease), progressive supranuclear palsy, and Wilson's disease. Disorders particularly suitable for treatment with the compounds of the present invention include Parkinson's disease, mild cognitive impairment, dementia (including Alzheimer's disease, vascular dementia, Lewy body dementia, and FTD), Huntington's disease, and amyotrophic lateral sclerosis, and in particular, Parkinson's disease, mild cognitive impairment, and dementia (including Alzheimer's disease, vascular dementia, Lewy body dementia, and FTD).

[0170] (Pharmaceutical composition) Compounds of formula (I) and compounds of formula (IZ) will generally be administered as part of a pharmaceutical composition.

[0171] Accordingly, in a further aspect of the present invention, there is provided a pharmaceutical composition comprising a compound of formula (I) or a compound of formula (IZ), or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable excipient or carrier.

[0172] The composition can be formulated for administration by any route, for example, for parenteral administration including intravenous, intramuscular, subcutaneous, or intradermal; or for oral, rectal, nasal, topical administration (including transdermal, eye drops, pulmonary, buccal, and sublingual topical surface administration), or for vaginal administration.

[0173] More preferably, the composition is formulated for parenteral administration or for oral administration, topical administration to the skin (transdermal administration), or topical administration to the lung (by inhalation).

[0174] The composition can be prepared by combining the active agent as defined above with a carrier. Generally, the formulation is prepared by uniformly and intimately combining the active agent with a liquid carrier or a finely divided solid carrier or both, and then shaping the product, if necessary. The present invention extends to a method for preparing a pharmaceutical composition, which comprises integrating or combining a compound of formula (I) with a pharmaceutically acceptable excipient or carrier.

[0175] The formulations for oral administration in the present invention can be provided as individual units, such as capsules, sachets, or tablets, each containing a predetermined amount of the active agent; as a powder or granules; as a solution or suspension of the active agent in an aqueous liquid or a non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion; or as a bolus agent or the like.

[0176] In some cases, the composition can be formulated for delayed, slow, or controlled release of the compound of formula (I) or the compound of formula (IZ).

[0177] For compositions for oral administration (e.g., tablets and capsules), the term "acceptable carrier" includes vehicles such as conventional excipients, for example, binders such as syrup, acacia, gelatin, sorbitol, tragacanth, polyvinylpyrrolidone (povidone), methylcellulose, ethylcellulose, sodium carboxymethylcellulose, hydroxypropylmethylcellulose, sucrose, and starch; fillers and carriers such as corn starch, gelatin, lactose, sucrose, microcrystalline cellulose, kaolin, mannitol, dicalcium phosphate, sodium chloride, and alginic acid; and lubricants such as magnesium stearate, sodium stearate, and other metal stearates, glycerol stearate, stearic acid, silicone fluid, talc wax, oils, and colloidal silica. Flavoring agents such as peppermint, wintergreen oil, cherry flavor, etc. can also be used. It may also be desirable to add a coloring agent to make the dosage form readily recognizable. Also, the tablets may be coated by methods well known in the art.

[0178] Tablets can be manufactured by compression or molding, optionally with one or more accessory ingredients. Compressed tablets can be prepared by compressing, in a suitable machine, a free-flowing form of the active agent, such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, preservative, surfactant, or dispersing agent. Molded tablets can be produced by molding, in a suitable machine, a mixture of the powdered compound moistened with an inert liquid diluent. The tablets may optionally be coated or scored and formulated to provide for slow or controlled release of the active agent.

[0179] Other formulations suitable for oral administration include lozenges containing the active agent in a flavored base, usually sucrose and acacia or tragacanth; pastilles containing the active agent in an inert base such as gelatin and glycerin, or sucrose and acacia; and mouthwashes containing the active agent in a suitable liquid carrier.

[0180] For topical surface application to the skin, the compound of formula (I) can be formulated into creams, ointments, jellies, solutions, suspensions, etc. Cream or ointment formulations that can be used in pharmaceuticals are conventional formulations well known in the art and are as described in standard pharmaceutical textbooks such as the British Pharmacopoeia.

[0181] Topical surface administration to the lung can be achieved by the use of aerosol formulations. Aerosol formulations generally contain the active ingredient suspended or dissolved in a suitable aerosol propellant, such as a chlorofluorocarbon (CFC) or a hydrofluorocarbon (HFC). Suitable CFC propellants include trichloromonofluoromethane (Propellant 11), dichlorotetrafluoromethane (Propellant 114), and dichlorodifluoromethane (Propellant 12). Suitable HFC propellants include tetrafluoroethane (HFC-134a) and heptafluoropropane (HFC-227). The propellant generally comprises 40% to 99.5%, for example 40% to 90% by weight, of the total inhalation composition. The formulation may contain excipients including co-solvents (such as ethanol) and surfactants (such as lecithin, sorbitan trioleate, etc.). Other possible excipients include polyethylene glycol, polyvinylpyrrolidone, glycerin, etc. The aerosol formulation is packaged in a canister and the appropriate dose is delivered by a metered valve (such as those supplied by Bespak, Valois, or 3M, or Aptar, Coster, or Vari).

[0182] Topical surface administration to the lung can also be achieved by the use of non-pressurized formulations, such as aqueous solutions or suspensions. These can be administered by nebulizers, which may be hand-held and portable or for use at home or in a hospital (i.e., non-portable). The formulation may contain excipients such as water, buffers, tonicity agents, pH adjusters, surfactants, and co-solvents. Suspensions and aerosol formulations (whether pressurized or non-pressurized) generally contain the compound of the present invention in a micronized form, for example with a D of 0.5 to 10 μm, for example about 1 to 5 μm. 50It may be included in a form having. The particle size distribution can be represented using the D 10 , D 50 , and D 90 values. The D 50 median of the particle size distribution is defined as the particle size in microns that divides the distribution in half. Measurements derived from laser diffraction are more accurately represented as a volume distribution, and as a result, the D 50 value obtained using this procedure is referred to as the Dv 50 value (median of the volume distribution). When used in this specification, the Dv value refers to the particle size distribution measured using laser diffraction. Similarly, the D 10 value and the D 90 value used in the case of laser diffraction are interpreted to mean the Dv 10 value and the Dv 90 value, respectively, where 10% of the distribution is below the particle size of the D 10 value, and 90% of the distribution is below the particle size of the D 90 value.

[0183] Local surface administration to the lung can also be achieved by the use of dry powder formulations. The dry powder formulation will contain the compound of the present disclosure in a micronized form, generally having a mass median aerodynamic diameter (MMAD) of 1 - 10 μm or a D 50 of 0.5 - 10 μm, for example about 1 - 5 μm. The powder of the compound of the invention in micronized form can be prepared by a micronization process or a similar size reduction process. Micronization can be carried out using a jet mill, for example one manufactured by Hosokawa Alpine. The resulting particle size distribution can be measured using laser diffraction (e.g., by a Malvern Mastersizer 2000S instrument). The formulation is generally a diluent acceptable for local surface use, such as lactose, glucose, or mannitol (preferably lactose), which usually has a relatively large particle size, for example a mass median aerodynamic diameter (MMAD) of 50 μm or more, for example 100 μm or more, or a D 50will include those having a size of 40 to 150 μm. As used herein, the term "lactose" refers to lactose-containing components and includes α-lactose monohydrate, β-lactose monohydrate, anhydrous α-lactose, anhydrous β-lactose, and amorphous lactose. The lactose component may be treated by micronization, sieving, grinding, compression, agglomeration, or spray drying. Commercially available forms of lactose in various forms also include, for example, Lactohale® (inhalation grade lactose; DFE Pharma), InhaLac® 70 (sieved lactose for dry powder inhalers; Meggle), Pharmatose® (DFE Pharma), and Respitose® (sieved inhalation grade lactose; DFE Pharma) products. In one embodiment, the lactose component is selected from the group consisting of α-lactose monohydrate, anhydrous α-lactose, and amorphous lactose. Preferably, the lactose is α-lactose monohydrate.

[0184] The dry powder formulation may also include other excipients. Thus, in one embodiment, the dry powder formulation of the present disclosure includes magnesium stearate or calcium stearate. Such formulations may have excellent chemical and / or physical stability, especially when such formulations also include lactose.

[0185] Dry powder formulations are generally delivered using a dry powder inhaler (DPI) device. Examples of dry powder delivery systems include SPINHALER®, DISKHALER®, TURBOHALER®, DISKUS®, SKYEHALER®, ACCUHALER®, and CLICKHALER®. Further examples of dry powder delivery systems include ECLIPSE, NEXT, ROTAHALER, HANDIHALER, AEROLISER, CYCLOHALER, BREEZHALER / NEOHALER, MONODOSE, FLOWCAPS, TWINCAPS, X-CAPS, TURBOSPIN, ELPENHALER, MIATHALER, TWISTHALER, NOVOLIZER, PRESSAIR, ELLIPTA, ORIEL dry powder inhaler, MICRODOSE, PULVINAL, EASYHALER, ULTRAHALER, TAIFUN, PULMOJET, OMNIHALER, GYROHALER, TAPER, CONIX, XCELOVAIR, and PROHALER.

[0186] In one embodiment, the compound of formula (I) is provided as a micronized dry powder formulation comprising, for example, a suitable grade of lactose.

[0187] Accordingly, as one aspect of the invention, there is provided a pharmaceutical composition comprising a compound of formula (I) or a compound of formula (IZ) in particulate form in combination with particulate lactose and optionally comprising magnesium stearate.

[0188] In one embodiment, the compound of formula (I) or the compound of formula (IZ) is provided as a micronized dry powder formulation comprising a suitable grade of lactose and magnesium stearate and filled into a device such as DISKUS. Preferably, such a device is a multidose device, for example, the formulation is filled into blisters for use in a multi-unit dose device such as DISKUS.

[0189] In another embodiment, the compound of formula (I) or the compound of formula (IZ) is provided as a micronized dry powder formulation, for example containing a suitable grade of lactose, and filled into hard shell capsules for use in single-dose devices such as AEROLISER.

[0190] In another embodiment, the compound of formula (I) or the compound of formula (IZ) is provided as a micronized dry powder formulation, containing a suitable grade of lactose and magnesium stearate, and filled into hard shell capsules for use in single-dose devices such as AEROLISER.

[0191] In another embodiment, the compound of formula (I) or the compound of formula (IZ) is provided as a fine powder for use in an inhaled dosage form, the powder being fine particles having a D 50 of 0.5 to 10 μm, for example about 1 to 5 μm, and is produced by a size reduction process other than jet milling micronization, for example spray drying, spray freezing, microfluidization, high pressure homogenization, supercritical fluid crystallization, ultrasonic crystallization, or a combination of these methods, or by other suitable particle formation methods known in the art used to produce fine particles having an aerodynamic particle size of 0.5 to 10 μm. The resulting particle size distribution can be measured using laser diffraction (e.g., by a Malvern Mastersizer 2000S instrument). The particles can contain the present compound alone or in combination with other suitable excipients that can assist in the processing. The resulting fine particles may form the final formulation for delivery to humans or, optionally, may be further formulated with other suitable excipients that promote delivery in an acceptable dosage form.

[0192] The compounds of the present invention may also be administered rectally in the form of suppositories or enemas, for example, aqueous or oily solutions, as well as suspensions, emulsions and foams. Such compositions are prepared according to standard procedures well known to those skilled in the art. For example, suppositories can be prepared by mixing the active ingredient with conventional suppository bases such as cocoa butter or other glycerides. In this case, the drug is mixed with a suitable non-irritating excipient that melts in the rectum and releases the drug because it is solid at room temperature but liquid at rectal temperature. Such materials are cocoa butter and polyethylene glycol.

[0193] Parenteral formulations are generally sterilized.

[0194] A physician, or other person skilled in the art, can determine the appropriate dosage of the compound of formula (I) or the compound of formula (IZ), and thus will be able to determine the amount of the compound of the present invention to be included in a particular pharmaceutical formulation (whether in unit dosage form or otherwise).

[0195] The compound of formula (I) or the compound of formula (IZ) can be used in combination with one or more other active agents useful for the treatment or prevention of neurodegenerative diseases, acute radiation syndrome or myalgic encephalomyelitis (ME, chronic fatigue syndrome), or post-viral syndrome (including chronic symptoms (long COVID) resulting from SARS-CoV2 infection).

[0196] Accordingly, in a further aspect of the present invention, there is provided a pharmaceutical composition further comprising an additional active agent useful for the treatment or prevention of neurodegenerative diseases, acute radiation syndrome or myalgic encephalomyelitis (ME, chronic fatigue syndrome) or post-viral syndrome (including chronic symptoms (long COVID) resulting from SARS-CoV2 infection) as described above.

[0197] There is also provided a product as a combined preparation for the simultaneous, sequential or separate use in the treatment or prevention of the above-mentioned neurodegenerative diseases, comprising a compound of formula (I) or a compound of formula (IZ), and an additional active agent useful for the treatment or prevention of neurodegenerative diseases.

[0198] A product as a combined preparation for simultaneous, sequential, or separate use in the treatment or prevention of acute radiation syndrome, comprising a compound of formula (I) or a compound of formula (IZ), and an additional active agent useful for the treatment or prevention of acute radiation syndrome, is likewise provided.

[0199] A product as a combined preparation for simultaneous, sequential, or separate use in the treatment or prevention of myalgic encephalomyelitis (ME, chronic fatigue syndrome) or post-viral syndrome (including chronic symptoms (long COVID) resulting from SARS-CoV2 infection), comprising a compound of formula (I) or a compound of formula (IZ), and an additional active agent useful for the treatment or prevention of ME or long COVID, is likewise provided.

[0200] (Figures and Examples) The present invention is further illustrated herein with reference to the following examples and figures.

Brief Description of the Drawings

[0201]

Figure 1

Figure 2

[0202] (General experimental procedure) Proton( 1 H) and carbon( 13 C) NMR spectra were recorded on a Bruker Avance(III)-500 spectrometer. Chemical shifts were referenced to the Me4Si (TMS, d 0) group, or the residual solvent peaks set at d 7.26 and 77.00 (CDCl3), or d 3.34 and 49.05 (CDOD), or d 2.50 and 39.43 (d6-DMSO) and recorded on a ppm scale with an internal standard. The reporting of NMR data is as follows: chemical shift (ppm), multiplicity (ap = apparent, s = singlet, d = doublet, t = triplet, q = quartet, sp = septet, br = broad, dd = double doublet, td = triple doublet, dt = double triplet, m = multiplet), coupling constant (Hz), integration.

[0203] Electrospray ionization (ESI) mass spectrometry (MS) experiments were performed on a QTOF Premier mass spectrometer (Micromass, UK) under normal conditions. A sodium formate solution was used as a calibration substance, and high-resolution mass spectra (HRMS) were measured. All reactions were monitored by thin-layer chromatography (TLC) using 0.2 mm silica gel (Merck Kieselgel 60 F 254)A pre-coated aluminum plate was used and visualized using a UV light, ammonium molybdate, ninhydrin, or potassium permanganate staining solution. Flash column chromatography was performed on a Grace Reveleris® automated flash system equipped with a continuous gradient device using Davisil® silica gel (60, particle size 0.040 - 0.063 mm), or Reveleris® silica, or a C-18 reverse-phase flash cartridge. Solvents for reactions and chromatography were of analytical grade and were used as received unless otherwise noted. Chiral and achiral high-performance liquid chromatography (HPLC) analyses were performed on an Agilent 1100 quaternary pump HPLC system equipped with a refractive index detector using the indicated columns. Data were processed with Agilent Cerity system software.

[0204] (Example 1) (tert-Butyl N-(benzyloxycarbonyl)-3-aza-7β-hydroxy-5β-cholan-24-oate (25a)) [Chemical formula]

[0205] (A. tert-Butyl 3α,7β-dihydroxy-5β-cholan-24-oate (12)) [Chemical formula] To a solution of ursodeoxycholic acid (10.2 g, 26.0 mmol) in dry THF (225 mL) was added trifluoroacetic anhydride (30.0 mL, 216 mmol) dropwise at 0 °C. After the addition was complete, the ice bath was removed and the reaction mixture was stirred for 1.5 h. Next, tert-butanol (63.8 mL) was introduced portionwise at room temperature and the reaction mixture was stirred overnight at room temperature. Then concentrated aqueous ammonia (53 mL) was added at room temperature and the reaction mixture was again stirred overnight. Saturated bicarbonate solution was added and the aqueous phase was extracted with ethyl acetate (3x). The combined organic fractions were washed with brine, dried over MgSO4 and concentrated. The crude product was purified by automated flash column chromatography (silica gel, ethyl acetate / petroleum ether 10 - 100%) to give quantitatively the product 12 as a colorless foam. [Chemical formula] HRMS (ESI) m / z, C 28 H 48 O4Na + Calculated as 471.3445, found 471.3440.

[0206] (B. tert-Butyl 7β-hydroxy-3-oxo-5β-cholan-24-oate (13)) [Chemical formula] To a solution of 12 (26.0 mmol), the product of step A, in dry dichloromethane (180 mL) was added (diacetoxyiodo)benzene (BAIB; 10.3 g, 32.0 mmol) and a catalytic amount of TEMPO (624 mg, 3.99 mmol). After stirring overnight, the reaction mixture was quenched with saturated Na2S2O3 solution and diluted with water. The organic layer was separated and the aqueous layer was further extracted with dichloromethane three times. The combined organic phases were washed with brine, dried over MgSO4 and concentrated. The residue was purified by automated flash column chromatography (silica gel, ethyl acetate / petroleum ether 2 - 100%) to give 11.4 g (98%) of the product 13 as a light yellow foam. [Chemical formula] HRMS(ESI) m / z, C 28 H 46 O4Na + Calculated value as 469.3288, measured value 469.3292.

[0207] (C. tert-Butyl 7β-hydroxy-4-oxa-3-oxo-4a-homo-5-cholan-24-oate (by-product 14a) and tert-butyl 7β-hydroxy-3-oxa-4-oxo-4a-homo-5-cholan-24-oate (desired product 15a))

Chem.

Chem.

[0208] (D. tert-Butyl 4,7β-dihydroxy-3,4-seco-5β-cholan-24-oate-3-amide (by-product 16a) and tert-butyl 2,7β-dihydroxy-2,3-seco-5β-cholan-24-oate-4-amide (desired product 17a))

Chem.

[0209] (tert - butyl 4,7β - dihydroxy - 3,4 - seco - 5β - cholan - 24 - oate - 3 - amide (16a))

Chem.

[0210] (tert-Butyl 2,7β-dihydroxy-2,3-seco-5β-cholan-24-oate-3-amide (17a))

Chemical formula

[0211] (tert-Butyl 4,7β-dihydroxy-3,4-seco-5β-cholan-24-oate-3-methyl ester (18a))

Chemical formula

[0212] (tert-Butyl 2,7β-dihydroxy-2,3-seco-5β-cholan-24-oate-3-methyl ester (19a))

Chemical formula

[0213] (E. tert-Butyl 2-amino-4,7β-dihydroxy-3-nor-3,4-seco-5β-cholan-24-oate (by-product, 20a) and tert-butyl 4-amino-2,7β-dihydroxy-3-nor-3,4-seco-5β-cholan-24-oate (desired product, 21a))

Chemical formula

[0214] (F. tert-Butyl N-(benzyloxycarbonyl)-2-amino-4,7β-dihydroxy-3-nor-3,4-seco-5β-cholan-24-oate (by-product 22a) and tert-butyl N-(benzyloxycarbonyl)-4-amino-2,7β-dihydroxy-3-nor-3,4-seco-5β-cholan-24-oate (desired product 23a)) [Chemical formula] A mixture of 20a and 21a (10.9 g, 24.1 mmol), the product of step E, was added to a 1:1 mixture of dichloromethane (160 mL) and water (160 mL), and sodium carbonate (13.1 g, 124 mmol) was added. The mixture was cooled to 0 °C. Next, benzyl chloroformate (CbzCl 95%; 4.00 mL, 28.0 mmol) was added dropwise, and the reaction mixture was stirred at 0 °C for 40 minutes. After the reaction was 28% complete (TLC analysis), aqueous ammonia (50 mL) was added, and the resulting mixture was diluted with water. The mixture was extracted with ethyl acetate (3x), and the combined organic phases were washed with brine, dried over MgSO4, and concentrated. The crude product mixture was purified by automated column chromatography (silica gel, ethyl acetate / petroleum ether 2 - 100%) to give 5.95 g (42%) of product 22a and 5.11 g (36%) of product 23a as colorless foams, respectively.

[0215] (tert-Butyl N-(benzyloxycarbonyl)-2-amino-4,7β-dihydroxy-3-nor-3,4-seco-5β-cholan-24-oate (22a))

Chem.

[0216] (tert-Butyl N-(benzyloxycarbonyl)-4-amino-2,7β-dihydroxy-3-nor-3,4-seco-5β-cholan-24-oate (23a))

Chem.

[0217] (G.tert-Butyl N-(benzyloxycarbonyl)-3-aza-7β-hydroxy-5β-cholan-25-oate (25a))

Chem.

[0218] (tert-Butyl N-(benzyloxycarbonyl)-3-aza-7β-hydroxy-5β-cholan-25-oate (25a)) [Chemical formula] HRMS (ESI) m / z, calculated for C 35 H 53 NO5Na + 590.3816, found 590.3815.

[0219] (tert-Butyl N-(benzyloxycarbonyl)-3-aza-7β-methanesulfonyloxy-5β-cholan-25-oate (26)) [Chemical formula] HRMS (ESI) m / z, calculated for C 36 H 55 NO7SNa + 668.3591, found 668.3590.

[0220] (Example 2) (tert-Butyl 3-aza-7β-hydroxy-5β-cholan-24-oate (27a))

Chem.

Chem.

[0221] (Example 3) (3-Aza-7β-hydroxy-5β-cholan-24-oic acid hydrochloride (28a))

Chem.

[0222] (Example 4) (tert-Butyl N-methyl-3-aza-7β-hydroxy-5β-cholan-24-oate (29a)) [Chemical formula] To a solution of the product 25a (203 mg, 0.358 mmol) from Example 1 in methanol (10 mL), formalin (37% aqueous solution; 0.15 mL, 2.02 mmol) was added, and the mixture was stirred at room temperature for 1 hour. Next, 10% palladium-on-carbon (37.8 mg) was added, and the atmosphere was exchanged with hydrogen. After stirring overnight, the reaction mixture was filtered through a celite plug, and the filtrate was concentrated. The crude product was purified by automated column chromatography [silica gel, (methanol: 28% aqueous ammonia solution 9:1) / ethyl acetate 0 - 20%] to obtain 161 mg (quantitative) of the product 29a as a colorless oil. [Chemical formula] HRMS(ESI) m / z, C 28 H 49 NO3H + Calculated value as 448.3785, measured value as 448.3782.

[0223] (Example 5) (N-Methyl-3-aza-7β-hydroxy-5β-cholan-24-oic acid hydrotrifluoroacetate (31a)) [Chemical formula] To a solution of the product 29a (93.0 mg, 0.208 mmol) of Example 4 in dry dichloromethane (5 mL) was added trifluoroacetic acid (4 mL) at 0 °C, and the resulting mixture was stirred for 2.5 h. The reaction mixture was then concentrated, and the crude product was purified by automated column chromatography [C18 silica gel, acetonitrile / water (+0.5% trifluoroacetic acid) 2 - 40%]. The purified product was evaporated at 60 °C with dichloromethane (3 - 4x) and methanol (1x) to give 51.1 mg (51%) of 31a as a colorless foam. [Chemical formula] HRMS (ESI) m / z, C 24 H 41 NO3H + Calculated as 392.3159, found 392.3169.

[0224] (Example 6) (N-Methyl-3-aza-7β-hydroxy-5β-cholan-24-oic acid hydrochloride (33)) [Chemical formula] To a solution of the product 29a (161 mg, 0.360 mmol) of Example 4 in dry dichloromethane (5 mL) was added trifluoroacetic acid (4 mL) at 0 °C. After stirring for 2.5 h, the reaction mixture was concentrated, and the residue was purified by automated column chromatography [C18 silica gel, acetonitrile / water (+0.5% trifluoroacetic acid) 2 - 40%]. The purified product was evaporated with dichloromethane (3 - 4x) and 3 M aqueous hydrochloric acid (3x). The resulting product was dried over potassium carbonate in a desiccator under high vacuum to give 112 mg (73%) of 33 as a colorless foam. [Chemical formula] HRMS (ESI) m / z, C 24 H 41 NO3H + Calculated as 392.3159, found 392.3174.

[0225] (Example 7) (tert-Butyl N-ethyl-3-aza-7β-hydroxy-5β-cholan-24-oate (30)) [Chemical formula] Using a method similar to that of Example 4, the compound (25a) (111 mg, 0.196 mmol) of Example 1 was dissolved in methanol (6 mL), treated with acetaldehyde (0.06 mL, 1.07 mmol), and then hydrogenated with 10% palladium on carbon (22.4 mg) to obtain 56.4 mg (63%) of the product 30 as a colorless foam. [Chemical formula] HRMS (ESI) m / z, C 29 H 51 NO3H + Calculated value as 462.3942, measured value 462.3942.

[0226] (Example 8) (N-Ethyl-3-aza-7β-hydroxy-5β-cholan-24-oic acid hydrotrifluoroacetate (32)) [Chemical formula] Using a method similar to that of Example 5, the compound (30) (50.0 mg, 0.108 mmol) of Example 7 was dissolved in dry dichloromethane (5 mL), reacted with trifluoroacetic acid (4 mL) at 0 °C for 2.5 hours to obtain 26.4 mg (47%) of the product 32 as a colorless foam. [Chemical formula] HRMS (ESI) m / z, C 25 H 43 NO3H + Calculated value as 406.3316, measured value 406.3325.

[0227] (Example 9) (tert-Butyl N-glycolyl-3-aza-7β-hydroxy-5β-cholan-24-oate (34a))

Chem.

Chem.

[0228] (Example 10) (N-Glycolyl-3-aza-7β-hydroxy-5β-cholan-24-oic acid (40a))

Chem.

[0229] (Example 11) (N-[(2S)-2-Amino-3-methylbutanoyl]-3-aza-7β-hydroxy-5β-cholan-24-oic acid hydrotrifluoroacetate (41a)) [Chemical formula]

[0230] (A. tert-Butyl N-{(2S)-2-[(tert-butoxycarbonyl)amino]-3-methylbutanoyl}-3-aza-7β-hydroxy-5β-cholan-24-oate (35a)) [Chemical formula] The same procedure as in Example 9 was adopted. The compound (27a) (100 mg, 0.231 mmol) of Example 2 was dissolved in dry dichloromethane (2.5 mL), and Boc-L-valine (Boc-Val-OH; 75.7 mg, 0.348 mmol), 1-hydroxybenzotriazole (HOBt; 34.6 mg, 0.256 mmol), N,N-diisopropylethylamine (0.089 mL, 0.511 mmol), and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDCI; 58.3 mg, 0.304 mmol) were sequentially added and treated. 122 mg (84%) of the product 35a was obtained as a colorless foam. [Chemical formula] HRMS(ESI) m / z, C 37 H 64 N2O6H + Calculated value as 633.4837, measured value 633.4839.

[0231] (B.N-[(2S)-2-Amino-3-methylbutanoyl]-3-aza-7β-hydroxy-5β-cholan-24-oic acid hydrotrifluoroacetate (41a)) [Chemical formula] The same deprotection method as in Example 10 was applied. The product (35a) (122 mg, 0.193 mmol) of Step A was dissolved in dry dichloromethane (5 mL) and treated with trifluoroacetic acid (4 mL) at 0 °C for 2.5 hours. The crude product was purified by automatic column chromatography [C18 silica gel, acetonitrile / water (+0.5% trifluoroacetic acid) 0 - 50%]. The purified product was evaporated at 60 °C with dichloromethane (3 - 4x) and methanol (1x), and 67.9 mg (60%) of 41a was obtained as a colorless foam. [Chemical formula] HRMS(ESI) m / z, C 28 H 48 N2O4H +Calculated value as 477.3687, measured value 477.3696.

[0232] (Example 12) (N-[(2S,3S)-2-Amino-3-methylpentanoyl]-3-aza-7β-hydroxy-5β-cholan-24-oic acid hydrotrifluoroacetate (42a))

Chemical formula

[0233] (A. tert-Butyl N-{(2S,3S)-2-[(tert-butoxycarbonyl)amino]-3-methylpentanoyl}-3-aza-7β-hydroxy-5β-cholan-24-oate (36a))

Chemical formula

Chemical formula

[0234] (B.N-[(2S,3S)-2-Amino-3-methylpentanoyl]-3-aza-7β-hydroxy-5β-cholan-24-oic acid hydrotrifluoroacetate (42a))

Chem.

Chem.

[0235] (Example 13) (N-[(2S)-2-Amino-4-methylpentanoyl]-3-aza-7β-hydroxy-5β-cholan-24-oic acid hydrotrifluoroacetate (43a))

Chem.

[0236] (A. tert-Butyl N-{(2S)-2-[(tert-butoxycarbonyl)amino]-4-methylpentanoyl}-3-aza-7β-hydroxy-5β-cholane-24-oate (37a))

Chem.

[0237] (B.N-[(2S)-2-amino-4-methylpentanoyl]-3-aza-7β-hydroxy-5β-cholan-24-oic acid hydrotrifluoroacetate (43a)) [Chemical formula] Following the same method as in Step B of Example 11, the compound (37a) (120 mg, 0.186 mmol) from Step A was dissolved in dry dichloromethane (5 mL) and stirred in the presence of trifluoroacetic acid (4 mL) until 73.3 mg (65%) of the product 43a was obtained as a colorless foam. [Chemical formula] HRMS(ESI) m / z, C 29 H 50 N2O4H + Calculated value as 491.3843, measured value 491.3857.

[0238] (Example 14) (N-[(2S)-2-Amino-3-(1H-indol-3-yl)propanoyl]-3-aza-7β-hydroxy-5β-cholan-24-oic acid hydrotrifluoroacetate (44a))

Chem.

[0239] (A. tert-Butyl N-{(2S)-2-[(tert-butoxycarbonyl)amino]-3-[1H-indol-3-yl]propanoyl}-3-aza-7β-hydroxy-5β-cholan-24-oate (38a))

Chem.

Chem.

[0240] (B. N-[(2S)-2-Amino-3-(1H-indol-3-yl)propanoyl]-3-aza-7β-hydroxy-5β-cholan-24-oic acid hydrotrifluoroacetate (44a))

Chem.

Chemical formula

[0241] (Example 15) (N-[(2S)-2-Amino-3-(4-hydroxyphenyl)propanoyl]-3-aza-7β-hydroxy-5β-cholan-24-oic acid hydrotrifluoroacetate (45a))

Chemical formula

[0242] (A. tert-Butyl N-{(2S)-2-[(tert-butoxycarbonyl)amino]-3-[4-hydroxyphenyl]propanoyl}-3-aza-7β-hydroxy-5β-cholan-24-oate (39a))

Chemical formula

Chemical formula

[0243] (B.N-[(2S)-2-amino-3-(4-hydroxyphenyl)propanoyl]-3-aza-7β-hydroxy-5β-cholan-24-oic acid hydrotrifluoroacetate (45a))

Chemical formula

Chemical formula

[0244] (Example 16) (tert-Butyl N-(benzyloxycarbonyl)-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oate (25b)) [ka]

[0245] (A. Methyl 3α,7β-3,7-dihydroxy-24-nor-5β-cholan-23-oate) The title compound was prepared from ursodeoxycholic acid by the method of D'Amore et al., 2014.

[0246] (B. Methyl 3α,7β-bis(tert-butyldimethylsilyloxy)-24-nor-5β-cholan-23-oate (3)) [ka] To a solution of the product of step A (diol 2; 37.8 g, 96.3 mmol) in dry DMF (1 L) was added imidazole (33.4 g, 491 mmol) and tert-butyl(chloro)dimethylsilane (60.3 g, 400 mmol) at room temperature. After stirring at 60 °C overnight, the reaction was diluted with water and extracted with ethyl acetate (3x). The combined organic layers were washed with water (2x) and brine, dried over MgSO4, and concentrated. The crude product was purified by flash column chromatography (silica gel, ethyl acetate / petroleum ether 1:19, 1:9, and 1:6) to give 53.6 g (90%) of product 3 as a colorless amorphous solid. [ka] HRMS(ESI)m / z, C 36 H 68 O4Si2Na + Calculated value 643.4548, measured value 643.4553. Procedure in Yan, S.; Ding, N.; Zhang, W.; Wang, P.; Li, Y.; Li, M. Carbohydr. Research 2012, 354, 6-20.

[0247] (C. 3α,7β-bis(tert-butyldimethylsilyloxy)-24-nor-5β-cholan-23-ol (4))

Chem.

Chem.

[0248] (D. 3α,7β-bis(tert-butyldimethylsilyloxy)-24-nor-5β-cholan-23-al (5))

Chem.

Chem.

[0249] (E. Methyl 3α,7β-bis(tert-butyldimethylsilyloxy)-25-homo-5β-cholan-23-ene-25-oate (7a) and tert-butyl 3α,7β-bis(tert-butyldimethylsilyloxy)-25-homo-5β-cholan-23-ene-25-oate (7b))

Chem.

[0250] (Methyl ester) To a solution of aldehyde 5 (8.48 g, 14.3 mmol), the product of step D, in dry dichloromethane (188 mL) was added methyl (triphenylphosphoranylidene) acetate (14.9 g, 44.6 mmol) at room temperature. After stirring overnight, the reaction mixture was concentrated to a small volume and directly loaded onto a silica gel-packed high-performance cartridge. Gradient elution with ethyl acetate and petroleum ether (0 - 15%) was performed using an automated chromatography system to obtain 8.74 g (94%) of the desired methyl ester product 7a as an amorphous colorless solid. [Chemical formula] HRMS (ESI) m / z, C 38 H 70 O4Si2Na + Calculated value as 669.4705, measured value 669.4708.

[0251] (tert-Butyl ester) The same procedure as for the preparation of methyl ester 7a was adopted. Aldehyde 5 (19.2 g, 32.5 mmol) was dissolved in dry dichloromethane (520 mL) and reacted with (tert-butoxycarbonylmethylene) triphenylphosphorane (43.0 g, 114 mmol) at room temperature for 72 hours to obtain 21.8 g (97%) of tert-butyl ester 7b as a colorless foam. [Chemical formula] HRMS (ESI) m / z, C 41 H 76 O4Si2Na + Calculated value as 711.5174, measured value 711.5175.

[0252] (F. Methyl 3α,7β-dihydroxy-25-homo-5β-cholan-25-oate (9a) and tert-butyl 3α,7β-dihydroxy-25-homo-5β-cholan-25-oate (9b)) [Chemical formula]

[0253] (Methyl ester) To a solution of alkene 7a (8.07 g, 12.5 mmol) which is the product of Step E in ethyl acetate (220 mL), 10% palladium on carbon (673 mg) was added and the atmosphere was exchanged with hydrogen. After stirring overnight at room temperature, the reaction mixture was filtered through celite and concentrated. The crude bis(tert-butyldimethylsilyloxy) reaction product 8a was redissolved in a mixture of THF (165 mL) and methanol (250 mL), and then 37% aqueous hydrochloric acid solution (57 mL) was added. After stirring for 2 hours at room temperature, the reaction mixture was carefully quenched with saturated bicarbonate solution and extracted with ethyl acetate (3x). The combined organic layers were washed with brine, dried over MgSO4 and then concentrated. The crude product was purified by automated column chromatography (silica gel, ethyl acetate / petroleum ether 2 - 100%) to give 4.69 g (89%) of the desired product 9a as a colorless foam.

[0254] (Methyl 3α,7β-bis(tert-butyldimethylsilyloxy)-25-homo-5β-cholan-25-oate (8a)) [Chemical formula] HRMS (ESI) m / z, C 38 H 72 O4Si2Na + Calculated value as 671.4861, measured value 671.4858.

[0255] (Methyl 3α,7β-dihydroxy-25-homo-5β-cholan-25-oate (9a)) [Chemical formula] HRMS (ESI) m / z, C 26 H 44 O4Na + Calculated value as 443.3132, measured value 443.3138.

[0256] (tert-Butyl ester) Using the same procedure as outlined for the preparation of methyl ester 8a, alkene 7b (21.7 g, 31.5 mmol) from step E was dissolved in ethyl acetate (400 mL) and hydrogenated overnight at room temperature with 10% palladium on carbon (867 mg) to afford 21.9 g (quantitative) of 8b as a colorless resin.

Chem.

[0257] A solution of silyl-protected tert-butyl ester 8b (21.6 g, 31.3 mmol) in THF (100 mL) was treated with a solution of tetrabutylammonium fluoride (144 mL, 144 mmol, 1 M in THF) at 50 °C for 72 h. After complete deprotection (TLC analysis), water was added and the aqueous phase was extracted with ethyl acetate (3x). The combined organic phases were washed with brine, dried over MgSO4 and concentrated. The crude product was purified by automated column chromatography (silica gel, ethyl acetate / petroleum ether 5 - 80%) to afford 13.9 g (96%) of the desired product 9b as a colorless oil.

Chem.

[0258] (G. tert-Butyl 7β-hydroxy-3-oxo-25-homo-5β-cholan-25-oate (11))

Chem.

[0259] (H.tert-butyl 7β-hydroxy-3-oxa-4-oxo-4a,25-bishomo-5β-cholan-25-oate (by-product 14b) and methyl 7β-hydroxy-4-oxa-3-oxo-4a,25-bishomo-5β-cholan-25-oate (desired product 15b)) [Chemical formula] Following the same procedure as outlined for Example 1C, the product from Step G (11) (11.9 g, 25.8 mmol) was treated with meta-chloroperbenzoic acid (mCPBA, 57 - 86%; 13.5 g, 54.8 mmol) in dry dichloromethane (200 mL) at room temperature overnight to afford 11.1 g (90%) of a 1:1 isomer mixture of products 14b and 15b as a colorless foam. [Chemical formula] HRMS(ESI) m / z, C 29 H 48 O5Na + (Mixture of isomers) Calculated value 499.3394, measured value 499.3398.

[0260] (I. tert-Butyl 4,7β-dihydroxy-25-homo-3,4-seco-5β-cholan-25-oate-3-amide (by-product 16b) and tert-butyl 2,7β-dihydroxy-25-homo-2,3-seco-5β-cholan-25-oate-3-amide (desired product 17b)) [Chemical formula] The same procedure as in Example 1D was carried out. The mixture of lactones 14b and 15b (10.9 g, 22.9 mmol) from step H was dissolved in dry 7N ammonia in methanol (155 mL) and heated overnight at 92 °C in a 200 mL sealed tube with a Teflon (trademark) screw cap (the sealed tube was filled to 4 / 5 of its volume and a blast shield was added), and 8.52 g (76%) of amide isomers 16b and 17b were obtained as a colorless foam. The product ratio (16b:17b) of 1:1.3 was determined by HPLC analysis (Phenomenex Luna C18(2) 5μm 250x4.6 mm; Phenomenex Security Guard C18 4x3 mm; mobile phase: 45:55:0.05 / acetonitrile / trifluoroacetic acid; flow rate: 1 mL / min; sample solvent: methanol; column temperature: 35 °C; injection volume: 25 μL; detection: refractive index). Furthermore, methyl ester isomers 18b (230 mg, 2%; colorless oil) and 19b (1.55 g, 13%, colorless foam) were isolated as by-products. Analytical samples of each of these four compounds were collected and subjected to spectroscopic property evaluation.

[0261] (tert-Butyl 4,7β-dihydroxy-25-homo-3,4-seco-5β-cholan-25-oate-3-amide (16b)) [Chemical formula] HRMS(ESI) m / z, C 29 H 51 NO5Na + Calculated value as 516.3659, measured value 516.3669.

[0262] (tert-butyl 2,7β-dihydroxy-25-homo-2,3-seco-5β-cholan-24-oate-3-amide (17b))

Chem.

[0263] (tert-butyl 4,7β-dihydroxy-25-homo-3,4-seco-5β-cholan-25-oate-3-methyl ester (18b))

Chem.

[0264] (tert-butyl 2,7β-dihydroxy-25-homo-2,3-seco-5β-cholan-25-oate-3-methyl ester (19b))

Chem.

[0265] (J. tert-Butyl 2-amino-4,7β-dihydroxy-25-homo-3-nor-3,4-seco-5β-cholan-25-oate (by-product 20b) and tert-butyl 4-amino-2,7β-dihydroxy-25-homo-3-nor-3,4-seco-5β-cholan-25-oate (desired product 21b))

Chem.

Chem.

[0266] (K. tert-Butyl N-(benzyloxycarbonyl)-2-amino-4,7β-dihydroxy-25-homo-3-nor-3,4-seco-5β-cholan-25-oate (by-product 22b) and tert-butyl N-(benzyloxycarbonyl)-4-amino-2,7β-dihydroxy-25-homo-3-nor-3,4-seco-5β-cholan-25-oate (desired product 23b))

Chem.

[0267] (N-(Benzyloxycarbonyl)-2-amino-4,7β-dihydroxy-25-homo-3-nor-3,4-seco-5β-cholan-25-oate (22b))

Chemical formula

[0268] (tert-Butyl N-(benzyloxycarbonyl)-4-amino-2,7β-dihydroxy-25-homo-3-nor-3,4-seco-5β-cholan-25-oate (23b))

Chemical formula

[0269] (L.tert-Butyl N-(benzyloxycarbonyl)-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oate (25b))

Chemical formula

[0270] (Example 17) (tert-Butyl 3-aza-7β-hydroxy-25-homo-5β-cholan-25-oate (27b)) [Chemical formula] Using the same procedure as described in Example 2, compound (25b) of Example 16 (865 mg, 1.48 mmol) was dissolved in methanol (35 mL) and hydrogenated with 10% palladium on carbon (93.8 mg) to give 675 mg (quantitative) of product 27b as a colorless foam. [Chemical formula] HRMS (ESI) m / z, C 28 H 49 NO3H + Calculated as 448.3785, found 448.3787.

[0271] (Example 18) (3-Aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid hydrochloride (28b)) [Chemical formula] The same protocol as in Example 3 was applied. The tert-butyl ester 27b (151 mg, 0.337 mmol) from Example 17 was dissolved in dry dichloromethane (4 mL) and deprotected with trifluoroacetic acid (TFA; 3 mL) at 0 °C to give 113 mg (78%) of 28b as a colorless foam. [Chemical formula] HRMS (ESI) m / z, calculated for C 24 H 41 NO3H + is 392.3159, found 392.3162.

[0272] (Example 19) (tert-Butyl N-methyl-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oate (29b)) [Chemical formula] Using the same procedure as in Example 4, the compound 25b (70.1 mg, 0.121 mmol) from Example 16 was dissolved in methanol (5 mL), treated with formalin (37% aqueous solution, 0.1 mL, 1.34 mmol), and stirred in a hydrogen atmosphere in the presence of 10% palladium on carbon (20.3 mg) to give 52.2 mg (94%) of 29b as a colorless oil. [Chemical formula] HRMS (ESI) m / z, calculated for C 29 H 51 NO3H + is 462.3942, found 462.3953.

[0273] (Example 20) (N-Methyl-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid hydrotrifluoroacetate (31b)) [Chemical formula] The same reaction protocol as in Example 5 was employed. The tert-butyl ester 29b (52.2 mg, 0.113 mmol) of Example 19 was dissolved in dry dichloromethane (5 mL), treated with trifluoroacetic acid (4 mL) at 0 °C, and 48.8 mg (83%) of the product 31b was obtained as a colorless foam.

Chemical formula

[0274] (Example 21) (tert-Butyl N-glycolyl-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oate (34b))

Chemical formula

Chemical formula

[0275] (Example 22) (N-Glycolyl-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid (40b))

Chem.

Chem.

[0276] (Example 23) (N-[(2S)-2-Amino-3-methylbutanoyl]-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid hydrotrifluoroacetate (41b))

Chem.

[0277] (A. tert-Butyl N-{(2S)-2-[(tert-butoxycarbonyl)amino]-3-methylbutanoyl}-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oate (35b))

Chem.

[0278] (B.N-[(2S)-2-Amino-3-methylbutanoyl]-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid hydrotrifluoroacetate (41b)) [Chemical formula] Following the same method as in Step B of Example 12, Compound 35b (137 mg, 0.212 mmol) of Step A was dissolved in dry dichloromethane (5 mL) and treated with trifluoroacetic acid (4 mL) to obtain 80.1 mg (63%) of Product 41b as a colorless foam. [Chemical formula] HRMS (ESI) m / z, C 29 H 50 N2O4H + Calculated value as 491.3843, measured value 491.3856.

[0279] (Example 24) (N-[(2S,3S)-2-Amino-3-methylpentanoyl]-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid hydrotrifluoroacetate (42b))

Chem.

[0280] (A.tert-Butyl N-{(2S,3S)-2-[(tert-butoxycarbonyl)amino]-3-methylpentanoyl}-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oate (36b))

Chem.

Chem.

[0281] (B.N-[(2S,3S)-2-Amino-3-methylpentanoyl]-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid hydrotrifluoroacetate (42b))

Chem.

Chemical formula

[0282] (Example 25) (N-[(2S)-2-Amino-4-methylpentanoyl]-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid hydrotrifluoroacetate (43b))

Chemical formula

[0283] (A. tert-Butyl N-{(2S)-2-[(tert-butoxycarbonyl)amino]-4-methylpentanoyl}-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oate (37b))

Chemical formula

Chemical formula

[0284] (B.N-[(2S)-2-Amino-4-methylpentanoyl]-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid hydrotrifluoroacetate (43b))

Chemical formula

Chemical formula

[0285] (Example 26) (N-[(2S)-2-Amino-3-(1H-indol-3-yl)propanoyl]-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid hydrotrifluoroacetate (44b))

Chemical formula

[0286] (A. tert-Butyl N-{(2S)-2-[(tert-butoxycarbonyl)amino]-3-[1H-indol-3-yl]propanoyl}-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oate (38b)) [Chemical formula] The same method as in Example 9 was applied. Compound 27b (49.8 mg, 0.111 mmol) of Example 17 was dissolved in dry dichloromethane (2 mL), and N α -Boc-L-tryptophan (51.3 mg, 0.169 mmol), 1-hydroxybenzotriazole (16.6 mg, 0.123 mmol), N,N-diisopropylethylamine (0.043 mL, 0.247 mmol), and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (30.4 mg, 0.159 mmol) were added for treatment, and 75.5 mg (93%) of product 38b was obtained as a colorless oil. [Chemical formula]

[0287] (B.N-[(2S)-2-amino-3-(1H-indol-3-yl)propanoyl]-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid hydrotrifluoroacetate (44b)) [Chemical formula] The same reaction conditions as in Step B of Example 11 were carried out. Compound 38b (75.5 mg, 0.103 mmol) of Step A was dissolved in dry dichloromethane (5 mL) and treated with trifluoroacetic acid (4 mL), and 47.6 mg (67%) of product 44b was obtained as a colorless foam. [Chemical formula] HRMS (ESI) m / z, C 35 H 51 N3O4H + Calculated value as 578.3952, measured value 578.3958.

[0288] (Example 27) (Conjugates of (3-Aza-7β-hydroxy-5β-cholan-24-oic acid (Compound 28a of Example 3) and 3-Aza-7β-hydroxy-25-homo-5β-cholan-24-oic acid (Compound 28b of Example 18)) (I. Preparation of N-(Benzyloxycarbonyl)-3-aza-7β-hydroxy-5β-cholan-24-oic acid (50a))

Chem.

Chem.

[0289] (II. Preparation of N-(Benzyloxycarbonyl)-3-aza-7β-hydroxy-24-homo-5β-cholan-25-oic acid (50b))

Chem.

[0290] (IIIa. General procedure A for conjugate formation) The Cbz-protected bile acid (1 equiv) from Step I (50a) or Step II (50b) was dissolved in anhydrous DMF (25 vol). HATU (2.0 equiv), DIPEA (3.0 equiv), and amine (1.2 equiv) were added, and the reaction was stirred at room temperature until completion as confirmed by TLC. After completion, the reaction mixture was diluted with H2O and extracted with ethyl acetate (×3). The combined organic layers were washed with saturated aqueous LiCl solution and concentrated. The crude residue was purified by column chromatography [C18 silica gel, gradient elution with H2O / MeOH] to give the conjugate as either the DIPEA salt or the free acid. The DIPEA salt was treated with a sodium ion exchange column to give the sodium salt of the desired residue.

[0291] (IIIb. General procedure B for conjugate formation) The Cbz-protected bile acid (1 equivalent) from Step I (50a) or Step II (50b) was dissolved in THF (25 vol) and cooled to 0 °C. Ethyl chloroformate (1.2 equivalents), followed by triethylamine (1.2 equivalents), was added, and the reaction mixture was stirred at 0 °C for 1 hour. After completion of the conversion of the starting material was confirmed by TLC, a solution of amine / thiol (1.5 equivalents) and NaHCO3 (1.5 equivalents) in H2O (25 vol) was added all at once, and then the mixture was stirred at 0 °C for 2 hours or until completion was confirmed by TLC. After completion, the mixture was concentrated under reduced pressure, diluted with H2O, acidified with 2M aqueous HCl to pH < 2. The suspension was extracted (ethyl acetate or THF; ×3), the organic layers were combined, washed with brine, dried over magnesium sulfate, and concentrated to give a crude residue. The crude material was purified by column chromatography.

[0292] (IV. General Procedure C for Deprotection of the Cbz Protecting Group) To a solution of the Cbz-protected conjugate (1 equivalent) from Step IIIa or Step IIIb in methanol (8 mL), 10% palladium on carbon (10 mol %) was added and the atmosphere was exchanged with hydrogen. The resulting reaction mixture was stirred at room temperature overnight. After completion of the conversion of the starting material was confirmed by TLC, the reaction mixture was filtered and concentrated under vacuum. The crude residue was purified by column chromatography [silica gel, gradient elution with ethyl acetate / methanol:triethylamine (9:1)] to give the conjugate as a colorless foam.

[0293] (N-(3-Aza-7β-hydroxy-5β-cholan-24-amido)-ethylsulfonic acid (Compound 51a)) [Chemical Structure] Using General Procedure A, Compound (50a) (50 mg, 0.098 mmol) was conjugated, and then using General Procedure C, it was deprotected to give Compound (51a) as a colorless glass (8.5 mg, 18%). [Chemical Structure] HRMS (ESI) m / z, C25 H 45 N2O5S + Calculated value as 485.3044, measured value 485.3053; TLC Rf 0.27.

[0294] (N-(3-Aza-7β-hydroxy-5β-cholan-24-amide)-acetic acid (Compound 52a))

Chem.

Chem.

[0295] (N-(3-Methyl-aza-7β-hydroxy-5β-cholan-24-oyl)-(2S)-2-amino-3-[(2-methylpropan-2-yl)oxy]propanoic acid (Compound 53a))

Chem.

Chem.

[0296] (N-(3-Methyl-aza-7β-hydroxy-5β-cholan-24-oil)-(R)-3-amino-3-phenylpropanoic acid (Compound 54a))

Chem.

Chem.

[0297] (N-(3-Methyl-aza-7β-hydroxy-5β-cholan-24-oil)-1-amino-4-fluorobenzene (Compound 55a))

Chem.

Chem.

[0298] (N-(3-Aza-7β-hydroxy-5β-cholan-24-oil)-morpholine (Compound 56a))

Chem.

Chemical formula

[0299] (N-(3-Aza-7β-hydroxy-5β-cholan-24-oil)-(S)-2-amino-3-hydroxypropanoic acid (Compound 57a))

Chemical formula

Chemical formula

[0300] (N-(3-Aza-7β-hydroxy-5β-cholan-24-oil)-3-amino-2-fluoropropanoic acid (Compound 58a))

Chemical formula

Chem.

[0301] (N-(Cyclohexyl)-N-(3-methyl-aza-7β-hydroxy-5β-cholan-24-yl)-cyclohexanamine (Compound 59a))

Chem.

[0302] (N-(3-Aza-7β-hydroxy-5β-cholan-24-yl)-4-aminobenzoic acid (Compound 60a))

Chem.

Chem.

[0303] (N-(3-Aza-7β-hydroxy-5β-cholan-24-yl)-(S)-2-amino-4-(methylthio)butanoic acid (Compound 61a))

Chemical formula

[0304] (N-(3-Aza-7β-hydroxy-5β-cholan-24-amide)-propanoic acid (Compound 62a))

Chemical formula

Chemical formula

[0305] (N-(3-Aza-7β-hydroxy-5β-cholan-24-yl)-(isopropyl-4-aminobenzoate) (Compound 63a))

Chemical formula

Chem.

[0306] (S-(3-Aza-7β-hydroxy-5β-cholan-24-yl)-phenylmethanethiol (Compound 64a))

Chem.

Chem.

[0307] (N-(3-Aza-7β-hydroxy-5β-cholan-24-yl)-(S)-2-amino-3-methylbutanoic acid (Compound 65a))

Chem.

Chem.

[0308] (N-(3-Aza-7β-hydroxy-5β-cholan-24-oil)-(2S,3S)-2-aminomethylpentanoic acid (Compound 66a))

Chem.

Chem.

[0309] (N-Methyl-N-(3-aza-7β-hydroxy-5β-cholan-24-oil)-glycine (Compound 67a))

Chem.

Chem.

[0310] (N-(3-Aza-7β-hydroxy-5β-cholan-24-oil)-(S)-2-aminopropanoic acid (Compound 68a)) [Chemistry] Using General Procedure B, the compound (50a) (40 mg, 0.078 mmol) was conjugated and then deprotected using General Procedure C to give the compound (68a) as a colorless glass (10.3 mg, 29%). [Chemistry] HRMS (ESI) m / z, C 26 H 45 N2O4 + Calculated value as 449.3374, measured value 449.3380; TLC Rf 0.18.

[0311] (N-(3-Aza-7β-hydroxy-25-homo-5β-cholan-25-yl)-(S)-2-aminobutanedioic acid (Compound 70b)) [Chemistry] Using General Procedure B, the compound (50b) (40 mg, 0.076 mmol) was conjugated and then deprotected using General Procedure C to give the compound (70b) as an off-white powder (7.5 mg, 19%). HRMS (ESI) m / z, C 28 H 47 N2O6 + Calculated value as 507.3429, measured value 507.3436; TLC Rf 0.04.

[0312] (N-(3-Aza-7β-hydroxy-25-homo-5β-cholan-25-yl)-(2S)-2-amino-3-[(2-methylpropan-2-yl)oxy]propanoic acid (Compound 71b)) [Chemistry] Using general procedure B, compound (50b) (40 mg, 0.076 mmol) was conjugated and then deprotected using general procedure C to give compound (71b) as an off-white powder (4.6 mg, 11%). [Chemical formula] HRMS(ESI) m / z, calculated value for C 31 H 55 N2O5 + is 535.4105, measured value is 535.4096; TLC Rf 0.24.

[0313] (N-(3-Aza-7β-hydroxy-25-homo-5β-cholan-25-amide)-ethylsulfonic acid (Compound 72b)) [Chemical formula] Using general procedure B, compound (50b) (40 mg, 0.076 mmol) was conjugated and then deprotected using general procedure C to give compound (72b) as a colorless glass (5.2 mg, 14%). [Chemical formula] HRMS(ESI) m / z, calculated value for C 26 H 47 N2O5S + is 499.3200, measured value is 499.3207; TLC Rf 0.25.

[0314] (N-(3-Aza-7β-hydroxy-25-homo-5β-cholan-25-oyl)-(S)-2-amino-3-phenylpropanoic acid (Compound 73b)) [Chemical formula] Using general procedure B, compound (50b) (40 mg, 0.076 mmol) was conjugated and then deprotected using general procedure C to give compound (73b) as an off-white powder (18.7 mg, 46%). [Chemical] HRMS(ESI) m / z, C 33 H 51 N2O4 + Calculated value as 539.3843, measured value 539.3849; TLC Rf 0.21.

[0315] (N-(3-Aza-7β-hydroxy-25-homo-5β-cholan-25-oil)-3-aminobutanoic acid (Compound 74b)) [Chemical] Using General Procedure B, compound (50b) (40 mg, 0.076 mmol) was conjugated, and then deprotected using General Procedure C to obtain compound (74b) as an off-white glassy solid (12.2 mg, 34%). [Chemical] HRMS(ESI) m / z, C 28 H 49 N2O4 + Calculated value as 477.3687, measured value 477.3678; TLC Rf 0.15.

[0316] (N-(3-Aza-7β-hydroxy-25-homo-5β-cholan-25-oil)-(S)-2-amino-4-methylpentanoic acid (Compound 75b)) [Chemical] Using General Procedure B, compound (50b) (40 mg, 0.076 mmol) was conjugated, and then deprotected using General Procedure C to obtain compound (75b) as a white powder (4.5 mg, 12%). [Chemical] HRMS(ESI) m / z, C 30 H 53 N2O4 +Calculated value as 505.4000, measured value 505.4007; TLC Rf 0.20.

[0317] (N-(3-Aza-7β-hydroxy-25-homo-5β-cholan-25-oil)-(2S)-2,6-diaminohexanoic acid (Compound 76b))

Chem.

Chem.

[0318] (N-(3-Aza-7β-hydroxy-25-homo-5β-cholan-25-oil)-(2S)-2-amino-3-(4-hydroxyphenyl)propanoic acid (Compound 77b))

Chem.

Chem.

[0319] (N-{(Benzyloxycarbonyl)-3-aza-7β-hydroxy-5β-cholan-24-oil}-2-oxa-6-azaspiro{3,3}heptane (Compound 78a))

Chem.

Chem.

[0320] (N-{(Benzyloxycarbonyl)-3-aza-7β-hydroxy-5β-cholan-24-oil}-4-piperidone (Compound 79a))

Chem.

Chem.

[0321] (N-{(Benzyloxycarbonyl)-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oil}-3-aminotetrahydrofuran (Compound 80b))

Chem.

Chem.

[0322] (N-{(3-Aza-7β-hydroxy-5β-cholan-25-yl}-isoindoline (Compound 81a))

Chem.

Chem.

[0323] (N-{(3-Aza-7β-hydroxy-25-homo-5β-cholan-25-yl)}-3-aminotetrahydrofuran (Compound 82b))

Chem.

Chem.

[0324] (Biological Example) (A. Primary Fibroblast Culture, Generation and Culture of iNPCs) Primary fibroblasts were continuously cultured in glucose-based medium (high glucose (4500 mg / L) Dulbecco's modified Eagle's medium (DMEM; Sigma-Aldrich) supplemented with 10% fetal bovine serum (Sigma-Aldrich), 100 IU / mL penicillin, 100 μg / mL streptomycin (Lonza), 1 mM sodium pyruvate (Sigma-Aldrich), and 50 μg / mL uridine (Sigma-Aldrich)). Unless otherwise specified, 24 hours before analysis, the glucose-based medium was replaced with galactose-based medium (glucose-free DMEM (Gibco) supplemented with the same content and further containing 5 mM galactose (Sigma-Aldrich)). All cell evaluations were performed at passages 6 - 10.

[0325] Induced neural progenitor cells (iNPCs) were generated according to a previous report (Meyer et al., "Direct conversion of patient fibroblasts demonstrates non-cell autonomous toxicity of astrocytes to motor neurons in familial and sporadic ALS", Proc Natl Acad Sci USA 2014). iNPCs were maintained in DMEM / Ham F12 (Invitrogen); N2, B27 supplement (Invitrogen), and FGFb (Peprotech) in fibronectin (Millipore)-coated tissue culture dishes and passaged routinely every 2 - 3 days using Accutase to dissociate them. This fibroblast cell line and iNPC cell line have already been characterized and published in Carling et al., 2020.

[0326] (B. Dopaminergic neuron differentiation of iNPCs) Briefly, iNPCs are plated in 6-well plates and cultured for 2 days in DMEM / F-12 medium supplemented with 1% NEAA, 2% B27 (Gibco), and Glutamax™ supplemented with 2.5 μM DAPT. On the third day, DAPT is removed and the medium is further supplemented with 1 μM smoothened agonist (SAG) and FGF8 (75 ng / ml) for an additional 10 days. Neurons are replated at this stage. Next, SAG and FGF8 are withdrawn and replaced with BDNF (30 ng / ml), GDNF (30 ng / ml), TGF-b3 (2 mM), and dcAMP (2 mM, Sigma) for 15 days. These are as previously reported in the literature by Schwartzentruber et al., 2020 and Carling et al., 2020. Dopaminergic neurons are treated with the compound at dosing concentrations of 0.1 nM, 1 nM, 3 nM, 10 nM, 30 nM, and 100 nM every 3 days for the last 12 days of differentiation.

[0327] (C. Immunofluorescence staining) Cells are plated in 96-well plates and fixed for 30 minutes using 4% paraformaldehyde. After washing with PBS, the cells are permeabilized for 10 minutes using 0.1% Triton™ X-100 and blocked for 1 hour using 5% goat serum. The cells are incubated with primary antibodies βIII tubulin (Millipore); activated caspase 3 (Cell Signaling) at 4 °C for 16 hours. The cells are washed using PBS-Tween® and incubated with Alexa Fluor®-conjugated secondary antibodies 488 and 568 (Invitrogen), and Hoescht (Sigma) 1 μM before imaging. Imaging was performed using an Opera Phenix™ high-content imaging system (Perkin Elmer).

[0328] (MMP protocol) Fibroblasts were cultured and plated at a density of 1000 cells per well in a Greiner black 384 μClear® plate with a medium volume of 50 μl. The plate was placed in an incubator overnight to allow the fibroblasts to attach to the plate surface. The next morning, the glucose-based medium was replaced with 25 μl of galactose-based medium. Next, compounds were administered to the plate using an ECHO® 550 liquid handling system. The wells were dosed to give a concentration range of 8 points from 0.06 nM to 300 nM of the compound. After dosing, the wells were filled with an additional 25 μl of galactose-based medium and then placed in the incubator for 24 hours. After 24 hours, the medium was removed from the wells and replaced with 25 μl of phenol-free minimum essential medium supplemented with 10% FBS, 1% penicillin-streptomycin, 1% sodium pyruvate, 0.1% uridine, 1% non-essential amino acids, and 1% MEM vitamins together with 80 nM TMRM (Sigma) and 10 μM Hoechst Stain (Sigma). One hour after returning the plate to the incubator, the staining medium was removed and replaced with 25 μl of phenol-free MEM. The plate was then imaged in 2 channels with 10 fields per well using an IN Cell high content microscope (GE Healthcare) (Cy3 excitation 542 nm, emission 604 - 64 nm; and DAPI excitation 350 nm, emission 450 - 55 nm, 37 °C, using CO2). After imaging, the plate was disposed of and the images were data mined using the IN Cell developer toolkit (GE Healthcare).

[0329] (Full ATP protocol) The ATP protocol is generally as described in the literature of Mortiboys et al., 2008. Briefly, fibroblasts were cultured and plated in a white 384-well plate at a concentration of 5000 cells per well with a media volume of 50 μl. The plate was placed in an incubator overnight to allow the fibroblasts to attach to the plate surface. The next morning, the glucose-based media was replaced with 25 μl of galactose-based media. Next, compounds were administered to the plate using an ECHO® 550 liquid handling system. The wells were dosed to give an 8-point concentration range of 0.06 nM to 300 nM of the compound. After dosing, the wells were filled with an additional 25 μl of galactose-based media and then placed in the incubator for 24 hours. After this incubation, the media was removed from the plate and the wells were washed twice with sterile PBS. The wells were filled with 25 μl of sterile PBS and then 12.5 μl of the Lysis solution of the ATPlite™ Luminescent ATP Detection Assay System (Perkin Elmer), including 16 cell-free wells used as a blank control. The plate was then placed on a rotary shaker at 700 rpm for 5 minutes. After shaking, 12.5 μl of the ATP substrate solution (Perkin Elmer) was added to each well and shaken for an additional 5 minutes. The plate was then placed in the dark for 10 minutes before reading the measurements. The luminescence intensity was recorded using a PHERAStar® plate reader. Immediately after the ATP assay, the DNA content of the plate was assayed with a CyQUANT® assay.

[0330] Immediately after the ATP assay, the DNA content was assayed with a CyQUANT® NF Cell Proliferation Assay Kit (ThermoFisher). The CyQUANT® buffer was prepared immediately before the assay and contained 1 μl of the CyQUANT® dye per ml of x1 HBSS solution. 12.5 μl of the CyQUANT® buffer was added to each well. The plate was placed in the incubator for 1 hour and then the excitation at 497 nm and the luminescence at 520 nm were read with a PHERAStar® plate reader.

[0331] Quantify the ATP in each well using the following formula: [Number] and determine using.

[0332] (Data analysis of the primary screening assay) After repeating the assay in triplicate for each line and compound, the data was entered into the Graph pad Prism 7 software suite to generate a dose-response curve using the default "[agonist] vs response (3 parameters)" equation. [Number] This EC 50 value was used to adopt the minimum and maximum responses and calculate the geometric mean between the five different lines to be evaluated.

[0333] Results from compounds that showed ambiguous results with the "[agonist] vs response (3 parameters)" equation were excluded from the geometric mean calculation because including them introduced high variance.

[0334] (Preliminary ATP protocol) In some cases, EC 50A preliminary ATP protocol that is not determined was used. The preliminary ATP protocol is generally as described in the literature of Mortiboys et al., 2008. Briefly, fibroblasts were cultured and plated at a concentration of 4000 cells per well in a 384-well plate with a white transparent bottom and a medium volume of 50 μl. The plate was placed in an incubator overnight to allow the fibroblasts to attach to the plate surface. The next morning, the glucose-based medium was replaced with 25 μl of galactose-based medium. Next, compounds were administered to the cells at concentrations of 100 nM and 1 μM using an ECHO® 550 liquid handling system. After dosing, the wells were filled with an additional 25 μl of galactose-based medium and then placed in the incubator for 24 hours. After this incubation, the medium was removed from the plate and the wells were washed twice with sterile PBS. The wells were filled with 20 μl of sterile PBS and then 10 μl of Lysis solution of the ATPlite™ Luminescence ATP Detection Assay System (Perkin Elmer), including 14 cell-free wells used as a blank control. Then, the plate was placed on a rotary shaker at 700 rpm for 10 minutes. After shaking, 10 μl of ATP substrate solution (Perkin Elmer) was added to each well and shaken for an additional 5 minutes. Then, the plate was incubated in the dark for 10 minutes and the measured values were read. The luminescence intensity was recorded using a FLUOstar® Omega plate reader. Immediately after the ATP assay, the DNA content of the plate was assayed by a CyQUANT® assay.

[0335] Immediately after the ATP assay, the DNA content is assayed with a CyQUANT® NF Cell Proliferation Assay Kit (ThermoFisher). The CyQUANT® buffer is prepared immediately before the assay and contains 2 μl of CyQUANT® dye per ml of x1 HBSS solution. 10 μl of CyQUANT® buffer is added to each well. Next, the cell plate is placed in the incubator for 1 hour and then the excitation at 497 nm and the luminescence at 520 nm are read with a FLUOStar® plate reader.

[0336] The ATP quantification for each well is determined as described above.

[0337] (Respiration measurement) The oxygen consumption rate (OCR) was measured using an Agilent Seahorse mitochondrial stress test with a 24-well Agilent Seahorse XF analyzer (Agilent) device. Human fibroblasts were plated at a density of 60,000 cells per well. The cells were treated with 50 nM of the compound for 24 hours before measurement. The OCR was measured three times at each of the following stages: basal state, after addition of oligomycin (0.5 μM), after addition of FCCP (0.5 μM), and after addition of rotenone (1 μM). Next, the cell number was counted on the immobilized assay plate using a Hoechst dye (1 μM). The displayed data are normalized to the cell number.

[0338] (Complex I assay) Ex vivo mouse brains were homogenized in a buffer of 250 mM sucrose, 20 mM HEPES, 3 mM EDTA, pH 7.5 at 4°C. The homogenization was performed using a Dounce homogenizer for cortical samples and by repeatedly passing through a 0.5 mm syringe for isolated striatum. Next, the samples were incubated on ice for 20 minutes with 30 μl of the wash buffer of the AbCam colorimetric Complex I assay kit. Next, the samples were centrifuged at 13,000 rpm for 30 minutes. Triplicate samples per condition were blocked on an AbCam colorimetric Complex I assay kit plate for 3 hours using the kit blocking buffer. Next, the samples were washed three times using the kit wash buffer and then the kit assay buffer containing NADH and the colorimetric dye was added. The assay plate was read at 450 nm every 30 seconds for 50 minutes using a plate reader with a kinetic assay program.

[0339] (Measurement of mitochondrial function and morphology in D.i neurons) i neurons were treated every three days for the last 12 days of the differentiation protocol. Cells were plated in 96-well plates; for live imaging, cells were incubated with 80 nM tetramethlyrhodamine (TMRM), 1 μM LysoTracker® Green (Invitrogen), and 1 μM Hoechst Stain (Sigma) for 1 hour at 37 °C and then imaged using Opera Phenix™. Cellular ATP measurements were performed using the ATPlite kit (Perkin Elmer) according to the manufacturer's instructions. Mitochondrial reactive oxygen species production was evaluated using the mitochondrial NpFR2 (probe; gift from Dr. Liz New, University of Sydney, Australia) at 20 μM and Hoechst Stain at 1 μM for 30 minutes at 37 °C, then the dye was removed and the cells were imaged using Opera Phenix™. Images generated in this live imaging experiment were analyzed using Harmony® (Perkin Elmer software). The inventors developed a protocol for separating the nucleus, cell boundaries and processes, mitochondria, lysosomes, and autophagosomes. Only z-projected images collected from z-stacks were analyzed.

[0340] (Results) (Fibroblasts) Mitochondrial membrane potential was measured in fibroblasts from three patients with sporadic Parkinson's disease (sPD) treated with the compounds of the present invention. The results are shown in Table 1, where "Bottom" = the maximum response at the lowest dose (0.06 nM) of the compound and "Top" = the maximum response at the highest dose (300 nM) of the compound.

[0341] (Table 1 - Mitochondrial membrane potential data from fibroblasts of three sporadic PD patients) [Table 1] Note: sPD patients have an average 18% decrease in MMP compared to controls. Thus, a 118% increase in MMP from vehicle-treated sPD patient levels will restore MMP to control levels.

[0342] Cell ATP levels were measured in fibroblasts from three patients with sporadic Parkinson's disease treated with the compounds of the invention. The results are shown in Table 2, where "bottom" = the maximum response at the lowest dose of the compound (0.06 nM) and "top" = the maximum response at the highest dose of the compound (300 nM).

[0343] (Table 2 - Cell ATP level data from fibroblasts of three sporadic PD patients in the full ATP assay) [Table 2] Note: sPD fibroblasts have an average 24% decrease in cell ATP levels compared to controls. Thus, 124% of vehicle-treated sPD fibroblasts is an increase to control ATP levels.

[0344] The MMP assay and ATP assay described above, along with toxicity measurements, comprise a primary screen of the compounds in patient primary fibroblasts. When considering which compound is most active in the primary screen, all information including the EC50 value indicating potency and the maximum response % of both assays is taken into account; based on the combined activity, a professional biologist makes a determination regarding each compound.

[0345] Oxygen consumption data for fibroblast lines from three sporadic PD patients and three controls are shown in Figures 1A, 1B, and 1C, from which it can be seen that sPD fibroblasts when treated with vehicle show a 30% decrease in basal mitochondrial respiration, a 42% decrease in spare respiratory capacity, and a 23% decrease in ATP-linked respiration compared to the control cell line. Treatment with compound 28b increases basal respiration in sPD fibroblasts relative to control levels (*p<0.05). Similar increases are observed, although to a lesser extent, in ATP-linked respiration and spare respiratory capacity in sPD fibroblasts. Note that since compound 28b was administered at 50 nM, it is clear that compound 28b provided an increase in mitochondrial function measurements as measured by oxygen consumption.

[0346] The above data show the mitochondrial protective effect of the compound in primary fibroblasts from sPD patients, but the cell type mainly affected in PD is dopaminergic neurons. The following data show the results obtained from dopaminergic neurons from sPD patients. These cultures are approximately 96% dopaminergic neurons and currently this methodology is the only protocol for generating such pure dopaminergic cultures from patient cells (developed by Mortiboys at the University of Sheffield and published in the literature by Schwarztentruber et al., 2020, Carling et al., 2020), and thus this is the patient-derived model that most closely represents the neurons affected in PD.

[0347] Table 3 below shows the results of measurements of mitochondrial function and neuronal morphology in i-neurons from sPD patients relative to controls when untreated or treated with either UDCA, compound 28a, or compound 28b.

[0348] (Table 3 - i-neurons)

Table 3

[0349] The data in Table 3 clearly show that in addition to compound 28b improving neuron morphology and reducing apoptosis levels (measured by activated caspase 3 levels), it confers a protective effect on both mitochondrial parameters in sPD-derived dopaminergic neurons. Apoptosis is a major mechanism of cell death in dopaminergic neurons in culture and in PD patients.

[0350] (In vivo mouse data with compound 28b) Figure 2 shows that mitochondrial respiratory chain complex I activity increases in whole brain homogenates of wild-type mice after administering 2 mg / kg of compound 28b intravenously (IV) and collecting cerebral hemisphere samples (from 3 animals per group) at 1, 4, 8, 12, and 24 hours after injection. Complex I activity increased by 125% 1 hour after administration and remained elevated to a similar extent 4 and 8 hours after injection. At 12 hours after injection, complex I activity began to decrease again but was still 98% higher than the control. At 24 hours after injection, complex I activity was also 35% higher than the control.

[0351] (Preliminary assay of the compound of Example 27) Preliminary assays with additional compounds of the present invention were performed to measure cellular ATP levels in fibroblasts from 3 patients with sporadic Parkinson's disease. The results are shown in Table 4.

[0352] (Cellular ATP level data from fibroblasts of 3 sporadic PD patients in the preliminary ATP assay)

Table 4

[0353] The data in Table 4 show that several compounds, such as compounds 52a, 54a, 55a, 57a, 62a, 64a, 65a, 66a, 67a, 75b, 77b, 78a, 79a, and 80b, show particularly good recovery of ATP production at both concentrations, and compound 64a restored ATP production to the control level at 100 nM. Since ATP is the energy component of cells and is important for many processes necessary for the health of these cells, increasing ATP production is the key to extending the lifespan of nerve cells in neurodegenerative diseases. Therefore, as described above, since mitochondrial dysfunction is considered to play a similar role in acute radiation syndrome, myalgic encephalomyelitis, and long COVID, enhancing ATP function would also be beneficial for the treatment of these conditions.

[0354] (Reference material)

Table 5

Claims

1. A compound of general formula (I), or a salt or solvate thereof: 【Chemical 1】 (wherein R 1 is selected from H, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C(O)R 4 and -C(O)OR 5 and the alkyl, alkenyl, and alkynyl R 1 groups are optionally substituted with one or more substituents independently selected from OR 11a and N(R 11a )(R 11b ); where each R 4 and R 5 are independently selected from OR 14a N(R 14a )(R 14b ), NH 3 + C(O)N(R 14a )(R 14b ), SR 14 a 5- or 6-membered nitrogen-containing heterocycle, and a 6- to 14-membered aryl or 5- to 14-membered heteroaryl (the aryl and heteroaryl are optionally substituted with one or more substituents selected from OH, halo, NH 2 NO 2 S(O) 2 OH, C 1-6 alkyl, C 1-6 haloalkyl, O(C 1-6 alkyl), and O(C 1-6 haloalkyl)) and is optionally substituted with one or more substituents selected from C 1-6 alkyl; and R 14a and R 14b are each independently selected from H and C 1-6 alkyl; and where each R 11a and R 11b are independently selected from H and C 1-4 alkyl, R 2 is selected from O and OH, R 2 If is O, [chemical 2] is a double bond, and R 2 If is OH, [C3] is a single bond; R 3 C(O)OH, C(O)OR 16 , C(O)N(R 6 )-X 1 -R 7 , C(O)N(R 8 )(R 9 ), and C(O)SR 10 Selected from: R 16 is optionally substituted with one or more substituents selected from OH, halo, and phenyl; 1-8 alkyl, wherein the phenyl is selected from halo, NO 2 , C.N., S(O) 2 O.H., C. 1-6 Alkyl, C 1-6 Haloalkyl, O(C 1-6 Alkyl)O(C 1-6 haloalkyl), O(R 15a ), N(R 15a )(R 15b ), and C(O)N(R 15a )(R 15b Optionally substituted with one or more substituents selected from Here, R 15a and R 15b are H and C, respectively. 1-6 Alkyl, and C 1-6 haloalkyl; X 1 , Halo, OR 12a , S.R. 12a , N(R 12a )(R 12b )C(O)OR 12a , C(O)N(R 12a )(R 12b ), N(R 12a )-C(NH)-N(R 12a )(R 12b ), N(R 12a ), -C(N + H 2 ), -N(R 12a )(R 12b ), and C alkylene optionally substituted with one or more substituents selected from 6- to 14-membered aryl or 5- to 14-membered heteroaryl (aryl and heteroaryl groups are optionally substituted with one or more substituents selected from halo, C 1-6 alkyl, C 1-6 haloalkyl OR 13a , N(R 13a )(R 13b ), NO 2 ), S(O) 2 OH, and CN); 1-6 wherein R and R 12a are each independently selected from H and C 12b alkyl; 1-6 wherein R and R 13a are each independently selected from H, C 13b alkyl, and C 1-6 haloalkyl; 1-6 wherein R is selected from H, methyl, and ethyl; 6 wherein R is selected from C(O)OH, C(O)O-(C 7 alkyl), S(O) 1-6 OH, and S(O) 2 O-(C 2 alkyl); 1-6 wherein R is selected from H, C 8 alkyl, and 3- to 6-membered carbocyclic groups ((C 1-6 alkyl, OH, O-(C 1-6 alkyl), N(R 1-6 )(R 19a ), C 19b haloalkyl, and halo, optionally substituted with one or more substituents selected therefrom)); 1-6 wherein R and R 19a are each independently selected from H and C 19b alkyl; 1-6 ​R 9 is H, C 1-6 selected from alkyl, a 3- to 7-membered carbocyclic group, a 3- to 7-membered heterocyclic group, a 6- to 14-membered aryl, and a 5- to 14-membered heteroaryl; wherein the alkyl group is C 1-4 alkyl, OH, O-(C 1-4 alkyl), C 1-4 haloalkyl, O-(C 1-4 haloalkyl), halo, N(R 19a )(R 19b ), phenyl, a 3- to 7-membered carbocyclic, and a 3- to 7-membered heterocyclic, and is optionally substituted with one or more substituents selected therefrom; wherein R 19a and R 19b are each independently as defined above; wherein the carbocyclic and heterocyclic groups are C 1-4 alkyl, OH, O-(C 1-4 alkyl), C 1-4 haloalkyl, O-(C 1-4 haloalkyl), oxo, phenyl, benzyl, and halo, and are optionally substituted with one or more substituents selected therefrom (provided that in this case, the heteroatom of the heterocyclic group is not substituted with OH, O-(C 1-4 alkyl), or O-(C 1-4 haloalkyl)); and wherein the aryl and heteroaryl are halo, NO 2 , CN, S(O) 2 OH, C 1-6 alkyl, C 1-6 haloalkyl, O(R 15a ), N(R 15a )(R 15b ), C(O)N(R 15a )(R 15b ), C(O)OH, and C(O)O-(C 1-6 alkyl), and are optionally substituted with one or more substituents selected therefrom; wherein R 15a and R 15b are each H, C 1-6 alkyl, and C 1-6selected independently of haloalkyl; or R 8 and R 9 are combined with the nitrogen atom to which they are attached to form a 4- to 10-membered heterocyclic group (optionally containing one or more additional heteroatoms selected from O, N, and S, and being optionally substituted with one or more substituents selected from C 1-4 alkyl, OH, O-(C 1-4 alkyl), halo, C 1-4 haloalkyl, O-(C 1-4 haloalkyl), C(O)OH, C(O)O(C 1-4 alkyl), phenyl, benzyl, CN, N(R 15a )(R 15b ), C(O)N(R 15a )(R 15b ), and oxo), provided that the heteroatom of the heterocyclyl group is not substituted with CN, N(R 15a )(R 15b ), OH, O-(C 1-4 alkyl), or O-(C 1-4 haloalkyl)); wherein the alkyl group is optionally substituted with one or more groups selected from O-(C 1-4 alkyl), O-(C 1-4 haloalkyl), N(R 15a )(R 15b ), OH, and C 3-6 cycloalkyl); or R 8 and R 9 are combined with the nitrogen atom to which they are attached to form a 5- to 10-membered heteroaryl group (optionally containing one or more additional heteroatoms selected from N, O, and S, and being optionally substituted with halo, NO 2 , CN, S(O) 2 OH, C 1-6 alkyl, C 1-6 haloalkyl, O(R 15a ), N(R 15a )(R 15b ), C(O)OH, C(O)N(R 15a )(R 15b ), and C(O)O-(C 1-6is optionally substituted with one or more substituents selected from (alkyl)); wherein R 15a and R 15b are each independently as defined above; wherein the alkyl group is optionally substituted with one or more groups selected from OH and C 3-6 cycloalkyl; wherein when the heteroaryl group contains a non-aromatic ring, the non-aromatic ring may also be substituted with oxo; R 10 is C 1-6 alkyl optionally substituted with OH, halo, or phenyl, wherein the phenyl is halo, NO 2 , CN, S(O) 2 OH, C 1-6 alkyl, C 1-6 haloalkyl, O(R 15a ), N(R 15a )(R 15b ), and C(O)N(R 15a )(R 15b ), and is optionally substituted with one or more substituents selected from; wherein R 15a and R 15b are each independently as defined above; and n is 1 or 2). **Claim 2** Formula (IZ): **Chemical Formula 4** (wherein: R 1 is selected from H, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C(O)R 4 , and -C(O)OR 5 , and wherein the alkyl, alkenyl, and alkynyl R 1 groups are optionally substituted with one or more substituents independently selected from OR 11a and N(R 11a )(R 11b ); wherein each R 4 and R 5is independently, OR 14a , N(R 14a )(R 14b ), NH 3 + , C(O)N(R 14a )(R 14b ), SR 14 a protected OH group, a protected NH 2 group, a protected C(O)NH 2 group, a 5- or 6-membered nitrogen-containing heterocycle, and a 6- to 14-membered aryl or 5- to 14-membered heteroaryl (the aryl and heteroaryl are optionally substituted with one or more substituents selected from OH, halo, NH 2 , NO 2 , S(O) 2 OH, C 1-6 alkyl, C 1-6 haloalkyl, O(C 1-6 alkyl), and O(C 1-6 haloalkyl)) and is optionally substituted with one or more substituents selected from the group consisting of C 1-6 alkyl; and R 14a and R 14b are each independently selected from H and C 1-6 alkyl; and herein, each R 11a and R 11b are independently selected from H and C 1-4 alkyl; R 2 is selected from O and OH, where when R 2 is O, 【Chemical Formula 5】 is a double bond, and when R 2 is OH, 【Chemical Formula 6】 is a single bond; R 3 is C(O)OH, C(O)OR 16 , C(O)N(R 6 )-X 1 -R 7 , C(O)N(R 8 )(R 9 ), and C(O)S-R 10selected from; R 16 is selected from C 1-8 alkyl and benzyl; X 1 is halo, OR 12a SR 12a N(R 12a )(R 12b )C(O)OR 12a C(O)N(R 12a )(R 12b N(R 12a )-C(NH)-N(R 12a )(R 12b N(R 12a )-C(N + H 2 )-N(R 12a )(R 12b ) a protected OH group, a protected NH 2 group, a protected C(O)NH 2 group, and a C 1-6 alkylene optionally substituted with one or more substituents selected from aryl having 6 to 14 members or heteroaryl having 5 to 14 members (the aryl and heteroaryl groups are optionally substituted with one or more substituents selected from halo, C 1-6 alkyl, C 13a haloalkyl OR 13a N(R 13b )(R 2 NO 2 S(O) 1-6 OH, and CN); R 12a and R 12b are each independently selected from H and C 1-6 alkyl; R 13a and R 13b are each independently selected from H, C 1-6 alkyl, and C 1-6 haloalkyl; R 6 is selected from H, methyl, and ethyl; R 7 is C(O)OH, C(O)O-(C 1-6 alkyl), S(O) 2 OH, and S(O) 2O-(C 1-6 is selected from O-(alkyl); R 8 is selected from H, C 1-6 alkyl, and a 3- to 6-membered carbocyclic ring (C 1-3 alkyl, OH, O-(C 1-3 alkyl), and optionally substituted with one or more substituents selected from halo); R 9 is selected from C 1-6 alkyl, a 5- or 6-membered carbocyclic ring (C 1-3 alkyl, OH, O-(C 1-3 alkyl) and optionally substituted with one or more substituents selected from halo), and phenyl (halo, NO 2 , CN, S(O) 2 OH, C 1-6 alkyl, O(R 15a ), N(R 15a )(R 15b ), C(O)OH, and C(O)O-(C 1-4 alkyl) and optionally substituted with one or more substituents selected from); wherein R 15a and R 15b are each independently selected from H and C 1-6 alkyl; or R 8 and R 9 are combined with the nitrogen atom to which they are attached to form a 5- or 6-membered heterocyclic ring (optionally containing one or more additional heteroatoms selected from O, N, and S, and C 1-4 alkyl, OH, O-(C 1-3 alkyl), halo, C(O)OH and C(O)O(C 1-4 alkyl) and optionally substituted with one or more substituents selected from); R 10 is OH, halo, or C 1-6 alkyl optionally substituted with phenyl, where phenyl is halo, OH, C 1-4 alkyl, C 1-4 haloalkyl, O(C 1-4 alkyl), and O(C 1-4optionally substituted with one or more substituents selected from (haloalkyl); The compound according to claim 1, or a salt or solvate thereof, which is a compound of (where n is 1 or 2).

3. In the above formula, 【Chemical Formula 7】 is a single bond, and the compound is of formula (IA) or (IB): 【Chemical Formula 8】 (wherein R 1 , R 3 , and n are as defined above for formula (I) or formula (IZ)) The compound, salt or solvate according to claim 1 or 2.

4. In the above formula, 【Chemical Formula 9】 is a double bond, and the compound is of formula (IC): 【Chemical Formula 10】 (wherein R 1 , R 3 , and n are as defined above for formula (I) or formula (IZ)) The compound, salt or solvate according to claim 1 or 2.

5. R 1 is H, C 1-6 alkyl, C 2-6 alkenyl, or C 2-6 alkynyl, The compound, salt or solvate according to any one of claims 1 to 4.

6. R 1 is H, methyl, or ethyl, The compound, salt or solvate according to claim 5.

7. R 1 is -C(O)R 4 where R 4 is as defined in claim 1, The compound, salt or solvate according to any one of claims 1 to 4.

8. R 4 is OH, NH 2 , NH 3 + , and is optionally substituted with one or more substituents selected from OH, NH, and aryl having 6 to 14 members or heteroaryl having 5 to 14 members (the aryl and heteroaryl are optionally substituted with one or more substituents selected from OH and halo)C 1-6 alkyl, the compound, salt or solvate according to claim 7.

9. R 4 is OH, NH 2 , NH 3 + , phenyl (optionally substituted with one or more substituents selected from OH and halo), and nitrogen-containing heteroaryl group (selected from pyrrole, pyridine, and indole, and optionally substituted with one or more substituents selected from OH and halo) and is optionally substituted with one or more substituents selected from C 1-6 alkyl, the compound, salt or solvate according to claim 8.

10. R 1 is an amino acid residue or a salt thereof, the compound, salt or solvate according to claim 7.

11. R 1 is selected from residues of glycine, valine, isoleucine, leucine, tryptophan, and tyrosine, and salts of those amino acid residues, the compound, salt or solvate according to claim 10.

12. R 1 is -C(O)OR 5 wherein R 5 is as described in claim 1, the compound, salt or solvate according to any one of claims 1 to 4.

13. R 5 is OH, NH 2 , NH 3 +, C optionally substituted with one or more substituents selected from aryl having 6 to 14 members and heteroaryl having 5 to 14 members (the aryl and heteroaryl are optionally substituted as described in claim 1) 1-6 The compound, salt or solvate according to claim 12, selected from C alkyl.

14. R 5 is, C 1-6 The compound, salt or solvate according to claim 13, which is alkyl (for example, t-butyl), benzyl, or fluorenylmethyl.

15. R 1 The compound, salt or solvate according to claim 14, wherein R is C(O)O-benzyl (CBz).

16. Formula (ID), (IE), (IF), or (IG): 【Chemical 11】 (wherein R 1 , R 2 and R 3 are as described in claim 1) The compound, salt or solvate according to any one of claims 1 to 15.

17. The counterion Z selected from chloride salt, trifluoroacetate salt, mesylate salt, bromide salt, sulfate salt, and fumarate salt - , particularly the counterion Z selected from chloride and trifluoroacetate - The compound, salt or solvate according to claim 16, in which is present.

18. The compound, salt or solvate according to any one of claims 1 to 17, wherein n is 1.

19. The compound, salt or solvate according to any one of claims 1 to 17, wherein n is 2.

20. R 3 is C(O)OH or C(O)OR 16 wherein R 16 is C 1-8The compound, salt or solvate according to any one of claims 1 to 19, wherein it is alkyl or benzyl.

21. R 3 is C(O)OH or C(O)O(C 1-4 alkyl), the compound or its salt or solvate according to claim 20.

22. R 3 is C(O)N(R 6 )-X 1 -R 7 wherein R 6 , X 1 and R 7 are as defined in claim 1, the compound, salt or solvate according to any one of claims 1 to 19.

23. R 6 is H or methyl, the compound, salt or solvate according to claim 22.

24. X 1 is unsubstituted, the compound, salt or solvate according to claim 22 or 23.

25. X 1 is substituted with one or more substituents selected from halo, OR 12a , SR 12a , N(R 12a )(R 12b )C(O)OR 12a , phenyl, and 5- or 6-membered heteroaryl (wherein the phenyl and heteroaryl groups are optionally substituted with one or more substituents selected from halo, C 1-6 alkyl, C 1-6 haloalkyl OR 13a , N(R 13a )(R 13b ), NO 2 , S(O) 2 OH, and CN); R 12a and R 12b are each independently selected from H and C 1-6 alkyl; and R 13a and R 13bis each independently H, C 1-6 alkyl, and C 1-6 haloalkyl, the compound, salt or solvate according to claim 22 or 23. **Claim 26** X 1 is one or more substituents selected from halo, OH, O(C 1-4 alkyl), SH, S(C 1-4 alkyl), C(O)OH, C(O)O-(C 1-6 alkyl) and phenyl (optionally substituted with one or more substituents selected from OH, halo, O(C 1-3 alkyl), and O(C 1-3 haloalkyl)), the compound, salt or solvate according to claim 25. **Claim 27** X 1 is one or more substituents selected from fluoro, OH, methoxy, ethoxy, i - propyloxy, s - butyloxy, t - butyloxy, S - methyl, NH 2 , C(O)OH, phenyl, and phenyl substituted with OH, the compound, salt or solvate according to claim 26. **Claim 28** R 7 is C(O)OH, C(O)O-(C 1-3 alkyl), and S(O) 2 OH, especially C(O)OH and S(O) 2 OH, the compound, salt or solvate according to any one of claims 22 - 27. **Claim 29** C(O)N(R 6 )-X 1 -R 7 is C(O)NH-(CH 2 ) 2 -SO 2 OH (taurine conjugate), C(O)NH - CH 2 -C(O)OH (glycine conjugate), O - t - butyl - L - serine (where R 3 is C(O)NH - CH(CH 2 O t(is Bu)-C(O)OH), β-phenylalanine (R 3 is C(O)NH-CH(Ph)-CH 2 (is -C(O)OH), serine (R 3 is C(O)NH-CH(CH 2 OH)-C(O)OH), 3-amino-2-fluoropropionic acid (R 3 is C(O)NH-CH 2 CHF-C(O)OH), methionine (R 3 is C(O)NH-CH(CH 2 CH 2 SMe)-C(O)OH), β-alanine (R 3 is C(O)NH-CH 2 CH 2 (is -C(O)OH), valine (R 3 is C(O)NH-CH( i Pr)-C(O)OH), isoleucine (R 3 is C(O)NH-CH(CH[Me]CH 2 Me)-C(O)OH), sarcosine (R 3 is C(O)N(Me)-CH 2 (is -C(O)OH), alanine (R 3 is C(O)NH-CH(Me)-C(O)OH), aspartic acid (R 3 is C(O)NH-CH(CH 2 C(O)OH)-C(O)OH), phenylalanine (R 3 is C(O)NH-CH(CH 2 Ph)-C(O)OH), 3-aminobutyric acid (R 3 is C(O)NH-CH(Me)CH 2 (is -C(O)OH), leucine (R 3 is C(O)NH-CH( s Bu)-C(O)OH), lysine (R 3 is C(O)NH-(CH 2 ) 4 -CH(NH 2 )-C(O)OH), and tyrosine (R 3 is C(O)NH-C(CH 2The compound, salt or solvate according to claim 22, selected from (Ph-OH)-C(O)OH). **Claim 30** R 3 is C(O)N(R 8 )(R 9 ), where R 8 and R 9 are as defined in claim 1, the compound, salt or solvate according to any one of claims 1 to 19. **Claim 31** R 8 is selected from H, C 1-4 alkyl, cyclopentyl, or cyclohexyl (the cyclopentyl and cyclohexyl groups are optionally substituted with methyl, OH, methoxy or fluoro, but more preferably are unsubstituted), the compound, salt or solvate according to claim 30. **Claim 32** R 8 is selected from H, methyl, ethyl, unsubstituted cyclopentyl, and unsubstituted cyclohexyl, the compound, salt or solvate according to claim 31. **Claim 33** R 9 is C 1-6 alkyl, such as methyl, and is either unsubstituted or substituted with a 3- to 7-membered heterocyclyl group, particularly a 5- or 6-membered heterocyclyl group, such as morpholinyl, piperidinyl, piperazinyl, pyrrolidinyl, or tetrahydrofuryl (the heterocyclyl group is either unsubstituted or may be substituted as defined in claim 1), the compound, salt or solvate according to any one of claims 30 to 32. **Claim 34** R 3 is C(O)N(R 8 )(R 9 ), where R 8 is H and R 9 is C 1-6 alkyl, such as methyl, which may be unsubstituted or may be substituted as defined in claim 33, the compound, salt or solvate according to any one of claims 30 to 33.

35. R 9 is methyl substituted with a 5- or 6-membered heterocyclyl group, such as morpholinyl, piperidinyl, piperazinyl, pyrrolidinyl, or tetrahydrofuryl (the heterocyclyl group may be unsubstituted or substituted as described in claim 1), the compound, salt or solvate according to claim 34.

36. R 9 is selected from a 3- to 7-membered carbocyclic group, a 3- to 7-membered heterocyclyl group, phenyl, or a 5- or 6-membered heteroaryl (any of these is optionally substituted as described in claim 1), the compound, salt or solvate according to any one of claims 30 to 32.

37. R 9 is a carbocyclic group selected from cyclopentyl and cyclohexyl, the compound, salt or solvate according to claim 36.

38. R 8 and R 9 are both a 3- to 6-membered cycloalkyl ring, such as cyclohexyl, the compound, salt or solvate according to claim 39.

39. R 9 is a 5- or 6-membered heterocyclyl containing 1 to 3 heteroatoms, especially N and / or O, the compound, salt or solvate according to claim 36.

40. R 9 is a heterocyclyl group selected from pyrrolidinyl, tetrahydrofuryl, piperidinyl, piperazinyl, and morpholinyl, which is unsubstituted or substituted as described in claim 1, the compound, salt or solvate according to claim 39.

41. The heterocyclyl group is substituted with oxo, the compound, salt or solvate according to claim 39 or 40.

42. R 9The compound, salt or solvate according to claim 36, wherein it is an aryl group having 6 to 14 members.

43. R 9 The compound, salt or solvate according to claim 42, wherein it is phenyl or naphthyl.

44. R 9 The compound, salt or solvate according to claim 42 or 43, wherein it is unsubstituted.

45. R 9 is substituted with one or more substituents selected from halo, C(O)OH, and C(O)O-(C 1-4 alkyl), the compound, salt or solvate according to claim 43.

46. R 8 is H, the compound, salt or solvate according to any one of claims 43 to 45.

47. R 9 The compound, salt or solvate according to claim 39, wherein it is a 5- or 6-membered heteroaryl group.

48. R 9 is pyridyl, the compound, salt or solvate according to claim 47.

49. R 9 is unsubstituted, the compound, salt or solvate according to claim 47 or 48.

50. R 9 is substituted with one or more substituents selected from halo, C(O)OH, and C(O)O-(C 1-4 alkyl), the compound, salt or solvate according to claim 47 or 48.

51. R 8 is H, the compound, salt or solvate according to any one of claims 47 to 50.

52. R 8 and R 9The compound, salt or solvate according to claim 30, which, together with the nitrogen atom to which they are attached, combine to form a 4- to 10-membered heterocyclic group (optionally containing one or more additional heteroatoms selected from O, N, and S and optionally substituted with one or more substituents as described in claim 1).

53. The compound, salt or solvate according to claim 52, wherein the heterocyclic group includes a monocyclic ring, particularly a 4- to 7-membered ring, such as a 5- or 6-membered ring.

54. The compound, salt or solvate according to claim 53, wherein the heterocyclic group is selected from morpholine, piperidine, piperazine, pyrrolidine, thiazoline, isothiazoline, thiazolidine, isothiazolidine, oxazoline, isoxazoline, oxazolidine, isoxazolidine, pyrazoline and pyrazolidine, and the heterocyclic group is unsubstituted or substituted as described in claim 1.

55. The compound, salt or solvate according to claim 54, wherein the heterocyclic group is selected from piperidine, pyrrolidine, piperazine, morpholine and isothiazolidine, and the heterocyclic group is unsubstituted or substituted as described in claim 1.

56. The compound, salt or solvate according to claim 52, wherein the heterocyclic group may include two or more rings, particularly two rings, which may be fused or bridged or joined by a spiro bond.

57. The heterocyclic group is R 8 and R 9 The compound, salt or solvate according to claim 56, which does not contain additional heteroatoms such that the nitrogen atom to which R and R are attached is the only heteroatom within the heterocyclic group.

58. The compound, salt or solvate according to claim 56, wherein the heterocyclic group contains one or more additional heteroatoms selected from N, O and S, for example one additional heteroatom or two additional heteroatoms.

59. The compound, salt or solvate according to claim 56, wherein the heterocyclic group is 2-oxa-6-azaspiro{3,3}heptane (i.e., an azetidine ring spiro-bonded to an oxetane ring at the 3-position).

60. The compound, salt or solvate according to any one of claims 52 to 59, wherein the heterocyclic group is unsubstituted.

61. The heterocyclic group is C 1-4 alkyl, OH, O-(C 1-4 alkyl), halo, C 1-4 haloalkyl, O-(C 1-4 haloalkyl), C(O)OH, C(O)O(C 1-4 alkyl), benzyl, N(R 15a )(R 15b )(e.g., NH 2 ), and oxo (e.g., C 1-3 alkyl, OH, O-(C 1-3 alkyl), halo, benzyl, NH 2 ), and oxo), and is substituted with one or more substituents selected from the group consisting of: The compound, salt or solvate according to any one of claims 52 to 59.

62. The compound, salt or solvate according to claim 61, comprising a substituent selected from halo, C 1-4 alkyl, C 1-4 haloalkyl, OH, O(C 1-4 alkyl), benzyl, N(R 15a )(R 15b )(e.g., NH 2 ), and oxo on a ring carbon atom.

63. The compound, salt or solvate according to claim 62, wherein the heterocyclic group is a piperidone ring or a pyrrolidone ring.

64. The compound, salt or solvate according to claim 61 or 62, comprising a substituent selected from C 1-4 alkyl, C 1-4 haloalkyl and benzyl on a ring nitrogen atom.

65. The compound, salt or solvate according to claim 61 or 62, comprising a cyclic sulfur atom having one or two oxo substituents.

66. R 8 and R 9 together with the nitrogen atom to which they are attached combine to form a 5- to 10-membered heteroaryl group (optionally containing one or more additional heteroatoms selected from N, O, and S), and the heteroaryl group is unsubstituted or is substituted with one or more substituents selected from halo, NO 2 , CN, S(O) 2 OH, C 1-6 alkyl, C 1-6 haloalkyl, O(R 15a ), N(R 15a )(R 15b ), C(O)OH, and C(O)O-(C 1-6 alkyl), the compound, salt or solvate according to claim 30.

67. The compound, salt or solvate according to claim 66, wherein the heteroaryl group has a 5- or 6-membered monocyclic ring.

68. The compound, salt or solvate according to claim 67, wherein the heteroaryl group is pyrrole, imidazole, triazole or thiazole.

69. The compound, salt or solvate according to claim 66, wherein the heteroaryl group has two rings.

70. Both rings are aromatic and the nitrogen atom to which either R 8 and R 9 is attached is part of a 5-membered ring fused to a further aromatic ring or aromatic heterocycle; or One of the rings is partially saturated or fully saturated, the compound, salt or solvate according to claim 69.

71. The heteroaryl group is selected from indole and isoindole; or The heteroaryl group is the above R 8 and R 9 The compound, salt or solvate according to claim 70, comprising a saturated or partially saturated ring containing a nitrogen atom to which it is attached, and a second ring which is a 5- or 6-membered ring (e.g., phenyl, pyridyl, or pyrrolyl).

72. R 8 is H and R 9 is 4-fluorophenyl; R 8 and R 9 are each cyclohexyl; R 8 is H and R 9 is 4-benzoic acid or its C 1-4 alkyl ester, e.g., isopropyl-4-benzoate; and, in the case of the compound of formula (I): R 8 is H and R 9 is tetrahydrofuranyl, especially tetrahydrofuran-3-yl; R 8 and R 9 are combined with the N atom to which they are attached to form an isoindoline ring; R 8 and R 9 are combined with the N atom to which they are attached to form a morpholine ring; R 8 and R 9 are combined with the N atom to which they are attached to form a piperidine or pyrrolidine ring substituted with oxo, e.g., 4-piperidone or 3-pyrrolidone; R 8 and R 9 are combined with the N atom to which they are attached to form a pyrrole ring, the compound, salt or solvate according to claim 30.

73. R 3 is C(O)S-R 10 (wherein R 10 is as defined in claim 1), the compound, salt or solvate according to any one of claims 1 to 19.

74. R 10 is C alkyl optionally substituted with OH, halo, or phenyl 1-6 The compound, salt or solvate according to claim 73, wherein **Claim 75** R 10 The compound, salt or solvate according to claim 74, wherein R is benzyl **Claim 76** R 2 is OH, R 3 is C(O)OH, and the compound is a compound of formula (IH) or a salt of formula (IJ): **Chemical Formula 12** (wherein R 1 is as defined in claim 1) The compound, salt or solvate according to any one of claims 1 to 19 **Claim 77** R 1 is H, R 2 is OH, and n is 1, and the compound is a compound of formula (IK) or a salt of formula (IL) **Chemical Formula 13** (wherein R 3 is as defined above for formula (I) or formula (IZ)) The compound, salt or solvate according to any one of claims 20 to 75 **Claim 78** The following: tert-Butyl N-(benzyloxycarbonyl)-3-aza-7β-hydroxy-5β-cholan-24-oate (25a); tert-Butyl 3-aza-7β-hydroxy-5β-cholan-24-oate (27a); 3-Aza-7β-hydroxy-5β-cholan-24-oic acid (28a); tert-Butyl N-methyl-3-aza-7β-hydroxy-5β-cholan-24-oate (29a); N-Methyl-3-aza-7β-hydroxy-5β-cholan-24-oic acid (31a and 33); tert-Butyl N-ethyl-3-aza-7β-hydroxy-5β-cholan-24-oate (30); N-Ethyl-3-aza-7β-hydroxy-5β-cholan-24-oic acid (32); tert-Butyl N-glycolyl-3-aza-7β-hydroxy-5β-cholan-24-oate (34a); N-Glycolyl-3-aza-7β-hydroxy-5β-cholan-24-oic acid (40a); N-[(2S)-2-Amino-3-methylbutanoyl]-3-aza-7β-hydroxy-5β-cholan-24-oic acid (41a); N-[(2S,3S)-2-Amino-3-methylpentanoyl]-3-aza-7β-hydroxy-5β-cholan-24-oic acid (42a); N-[(2S)-2-Amino-4-methylpentanoyl]-3-aza-7β-hydroxy-5β-cholan-24-oic acid (43a); N-[(2S)-2-Amino-3-(1H-indol-3-yl)propanoyl]-3-aza-7β-hydroxy-5β-cholan-24-oic acid (44a); N-[(2S)-2-Amino-3-(4-hydroxyphenyl)propanoyl]-3-aza-7β-hydroxy-5β-cholan-24-oic acid (45a); tert-Butyl N-(benzyloxycarbonyl)-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oate (25b); tert-Butyl 3-aza-7β-hydroxy-25-homo-5β-cholan-25-oate (27b); 3-Aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid (28b); tert-Butyl N-methyl-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oate (29b); N-Methyl-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid (31b); tert-Butyl N-glycolyl-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oate (34b); N-Glycolyl-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid (40b); N-[(2S)-2-Amino-3-methylbutanoyl]-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid (41b); N-[(2S,3S)-2-Amino-3-methylpentanoyl]-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid (42b); N-[(2S)-2-Amino-4-methylpentanoyl]-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid (43b, Example 25); N-[(2S)-2-Amino-3-(1H-indol-3-yl)propanoyl]-3-aza-7β-hydroxy-25-homo-5β-cholan-25-oic acid (44b); N-(Benzyloxycarbonyl)-3-aza-7β-hydroxy-5β-cholan-24-oic acid (50a); N-(3-Aza-7β-hydroxy-5β-cholan-24-amido)-ethylsulfonic acid (51a); N-(3-Aza-7β-hydroxy-5β-cholan-24-amido)-acetic acid (52a); N-(3-Methyl-aza-7β-hydroxy-5β-cholan-24-oyl)-(2S)-2-amino-3-[(2-methylpropan-2-yl)oxy]propanoic acid (53a); N-(3-Methyl-aza-7β-hydroxy-5β-cholan-24-oyl)-(R)-3-amino-3-phenylpropanoic acid (54a); N-(3-Methyl-aza-7β-hydroxy-5β-cholan-24-oyl)-1-amino-4-fluorobenzene (55a); N-(3-Aza-7β-hydroxy-5β-cholan-24-oyl)-morpholine (56a); N-(3-Aza-7β-hydroxy-5β-cholan-24-oil)-(S)-2-amino-3-hydroxypropanoic acid (57a); N-(3-Aza-7β-hydroxy-5β-cholan-24-oil)-3-amino-2-fluoropropanoic acid (58a); N-(Cyclohexyl)-N-(3-methyl-aza-7β-hydroxy-5β-cholan-24-oil)-cyclohexanamine (59a); N-(3-Aza-7β-hydroxy-5β-cholan-24-oil)-4-aminobenzoic acid (60a); N-(3-Aza-7β-hydroxy-5β-cholan-24-oil)-(S)-2-amino-4-(methylthio)butanoic acid (61a); N-(3-Aza-7β-hydroxy-5β-cholan-24-amide)-propanoic acid (62a); N-(3-Aza-7β-hydroxy-5β-cholan-24-oil)-(isopropyl-4-aminobenzoate) (63a); S-(3-Aza-7β-hydroxy-5β-cholan-24-oil)-phenylmethanethiol (64a); N-(3-Aza-7β-hydroxy-5β-cholan-24-oil)-(S)-2-amino-3-methylbutanoic acid (65a); N-(3-Aza-7β-hydroxy-5β-cholan-24-oil)-(2S,3S)-2-amino-3-methylpentanoic acid (66a); N-Methyl-N-(3-aza-7β-hydroxy-5β-cholan-24-oil)-glycine (67a); N-(3-Aza-7β-hydroxy-5β-cholan-24-oil)-(S)-2-aminopropanoic acid (68a); N-(Benzyloxycarbonyl)-3-aza-7β-hydroxy-24-homo-5β-cholan-25-oic acid (50b); N-(3-Aza-7β-hydroxy-27-homo-5β-cholan-27-oil)-(S)-2-aminobutanedioic acid (70b); N-(3-Aza-7β-hydroxy-27-homo-5β-cholan-27-yl)-(2S)-2-amino-3-[(2-methylpropan-2-yl)oxy]propanoic acid (71b); N-(3-Aza-7β-hydroxy-27-homo-5β-cholan-27-amide)-ethylsulfonic acid (72b); N-(3-Aza-7β-hydroxy-27-homo-5β-cholan-27-yl)-(S)-2-amino-3-phenylpropanoic acid (73b); N-(3-Aza-7β-hydroxy-27-homo-5β-cholan-27-yl)-3-aminobutanoic acid (74b); N-(3-Aza-7β-hydroxy-27-homo-5β-cholan-27-yl)-(S)-2-amino-4-methylpentanoic acid (75b); N-(3-Aza-7β-hydroxy-27-homo-5β-cholan-27-yl)-(2S)-2,6-diaminohexanoic acid (76b); N-(3-Aza-7β-hydroxy-27-homo-5β-cholan-27-yl)-(2S)-2-amino-3-(4-hydroxyphenyl)propanoic acid (77b); N-{(Benzyloxycarbonyl)-3-aza-7β-hydroxy-5β-cholan-24-yl}-2-oxa-6-azaspiro{3,3}heptane (78a); N-{(Benzyloxycarbonyl)-3-aza-7β-hydroxy-5β-cholan-24-yl}-4-piperidone (79a); N-{(Benzyloxycarbonyl)-3-aza-7β-hydroxy-25-homo-5β-cholan-25-yl}-3-aminotetrahydrofuran (80b); N-{(3-Aza-7β-hydroxy-5β-cholan-25-yl}-isoindoline (81a); N-{(3-Aza-7β-hydroxy-25-homo-5β-cholan-25-yl)}-3-aminotetrahydrofuran (82b); The compound according to claim 1 selected from, and salts or solvates thereof.

79. A.R 1 is C(O)OR 5 and R 3 is C(O)OR 16 For a compound of formula (I) wherein: 【Chemical 14】 (wherein R 2 and n are as defined for formula (I), and R 1a is C(O)OR 5 and R 3a is C(O)OR 16 and wherein R 5 and R 16 are as defined for formula (I)), reacting the compound with methanesulfonyl chloride in an organic solvent, such as pyridine, at a low temperature, such as about -5 to 5 °C; B.R 1 For a compound of formula (I) wherein B.R 1 is C(O)OR 5 hydrogenating the compound of formula (I) with a palladium catalyst; C.R 1 is C 1-6 alkyl and R 3 is C(O)OR 16 For a compound of formula (I) wherein R 1 is H and R 3 is C(O)OR 16 reacting the compound of formula (I) with a compound of formula (XX): R 1b -C(O)H (XX) (wherein R 1b is C 1-5 alkyl), and then hydrogenating with a palladium / carbon catalyst; D. R 1 is C 2-6 alkenyl or C 2-6 alkynyl and R 3 is C(O)OR 16 For a compound of formula (I) wherein R 1 is H and R 3 is C(O)OR 16 reacting the compound of formula (I) with a compound of formula (XVIII): R 1c-X (XVIII) (wherein R 1c is C 2-6 alkenyl or C 2-6 alkynyl, and X is a leaving group such as halo, for example chloro or bromo) with a compound in the presence of a base (wherein when R 2 is OH, further including a first step of protecting R 2 with a suitable protecting group and a final step of removing the protecting group); E. For a compound in which E.R 3 is C(O)OH, hydrolyzing a compound of formula (I) in which R 3 is C(O)OR 16 with an acid or a base; F. For a salt of formula (I) in which the nitrogen atom to which R 1 is attached is quaternized, treating a compound of formula (I) in which R 1 is H with an acid, for example hydrochloric acid or trifluoroacetic acid; G. For a compound of formula (I) in which R 1 is C(O)R 4 and R 3 is C(O)OR 16 : (i) A compound of formula (I) in which R 1 is H and R 3 is C(O)OR 16 with formula (XXI): R 4 '-C(O)OH (XXI) (wherein R 4 ' is OR 14a , N(R 14a )(R 14b ), NH 3 + + 14a ), C(O)N(R 14b )(R 14 ), a protected OH group, a protected NH 2 group, a protected C(O)NH 2 group, a 5- or 6-membered nitrogen-containing heterocycle, and a 6- to 14-membered aryl or 5- to 14-membered heteroaryl (the aryl and heteroaryl are OH, halo, NH 2 , NO 2, S(O) 2 OH, C 1-6 alkyl, C 1-6 haloalkyl, O(C 1-6 alkyl), and O(C 1-6 haloalkyl) optionally substituted with one or more substituents selected from) optionally substituted with one or more substituents selected from C 1-6 alkyl; and reacting the compound with a coupling reagent under basic conditions, for example using N,N - diisopropylethylamine, and in the presence of a coupling reagent; and (ii) removing the protecting group if necessary to obtain a group R 2 containing OH, NH 2 or C(O)NH 4 groups; H. R 1 is C(O)R 4 and R 3 is C(O)OH, reacting the product of G(i) with trifluoroacetic acid for the compound of formula (I); I. R 3 is C(O)N(R 6 )-X 1 -R 7 for the compound of formula (I), reacting the compound of formula (I) wherein R 3 is C(O)OH with a compound of formula (XXV): HN(R 6 )-X 1 -R 7 (XXV) (wherein R 6 , R 7 and X 1 are as defined for formula (I)) in the presence of a coupling reagent and under basic conditions such as in the presence of an amine, for example diisopropylethylamine (DIPEA) or triethylamine (TEA), in an organic solvent such as DMF; J. R 3 is C(O)N(R 8 )(R 9 ) for the compound of formula (I), reacting the compound of formula (I) wherein R 3 is C(O)OH with a compound of formula (XXVI): HN(R 8)(R 9 ) (XXVI); (wherein R 8 and R 9 are as defined for formula (I)) with a compound under basic conditions and in the presence of a coupling agent; K. R 8 and R 9 are combined with the nitrogen atom to which they are attached to form a 5-membered heteroaryl ring for a compound of formula (I) where R 3 is C(O)N(R 8 )(R 9 )(wherein R 8 and R 9 are combined with the nitrogen atom to which they are attached to form a 5- or 6-membered heterocyclic ring substituted with oxo), to subject the compound of formula (I) to a coupling reaction; L. R 3 is C(O)S-R 10 for a compound of formula (I), reacting a compound of formula (I) where R 3 is C(O)OH with a compound of formula (XXVII): HSR 10 (XXVII) (wherein R 10 is as described in claim 1), a process for preparing a compound, salt or solvate according to any one of claims 1 to 78.

80. A compound, salt or solvate according to any one of claims 1 to 78 for use in medicine.

81. A compound, salt or solvate according to any one of claims 1 to 78 for use in the treatment or prevention of neurodegenerative diseases.

82. The neurodegenerative disease is selected from Parkinson's disease, mild cognitive impairment, dementia (including Alzheimer's disease, vascular dementia, Lewy body dementia and frontotemporal dementia (FTD)), Huntington's disease, amyotrophic lateral sclerosis (motor neuron disease), progressive supranuclear palsy, and Wilson's disease, a compound, salt or solvate for use according to claim 81.

83. A compound, salt or solvate according to any one of claims 1 to 78 for use in the treatment or prevention of acute radiation syndrome.

84. A compound, salt or solvate according to any one of claims 1 to 78 for use in the treatment or prevention of myalgic encephalomyelitis (ME, chronic fatigue syndrome) or post-viral syndrome, including chronic symptoms resulting from SARS-CoV2 infection (long COVID).

85. Use of a compound, salt or solvate according to any one of claims 1 to 78 in the preparation of a therapeutic agent for neurodegenerative diseases, acute radiation syndrome or myalgic encephalomyelitis (ME, chronic fatigue syndrome) or post-viral syndrome (including chronic symptoms resulting from SARS-CoV2 infection (long COVID)).

86. A method for the treatment or prevention of neurodegenerative diseases, acute radiation syndrome or myalgic encephalomyelitis (ME, chronic fatigue syndrome) or post-viral syndrome (including chronic symptoms resulting from SARS-CoV2 infection (long COVID)), the method comprising administering to a patient in need thereof an effective amount of a compound, salt or solvate according to any one of claims 1 to 78.

87. A pharmaceutical composition comprising a compound, salt or solvate according to any one of claims 1 to 78 and a pharmaceutically acceptable excipient or carrier.

88. The pharmaceutical composition according to claim 87 formulated for parenteral administration or for oral administration, topical administration to the skin (transdermal administration) or topical administration to the lung (by inhalation).

89. A process for preparing the pharmaceutical composition according to claim 87 or 88, comprising combining a compound, salt or solvate according to any one of claims 1 to 78 with a pharmaceutically acceptable excipient or carrier.

90. A product as a combined preparation for simultaneous, sequential, or separate use in the treatment or prevention of a neurodegenerative disease, comprising a compound, salt or solvate according to any one of claims 1 to 78, and an additional active agent useful for the treatment or prevention of a neurodegenerative disease.

91. A product as a combined preparation for simultaneous, sequential, or separate use in the treatment or prevention of acute radiation syndrome, comprising a compound, salt or solvate according to any one of claims 1 to 78, and an additional active agent useful for the treatment or prevention of acute radiation syndrome.

92. A product as a combined preparation for simultaneous, sequential, or separate use in the treatment or prevention of myalgic encephalomyelitis (ME, chronic fatigue syndrome) or post-viral syndrome (including chronic symptoms (long COVID) resulting from SARS-CoV2 infection), comprising a compound, salt or solvate according to any one of claims 1 to 78, and an additional active agent useful for the treatment or prevention of myalgic encephalomyelitis (ME, chronic fatigue syndrome) or post-viral syndrome (including chronic symptoms (long COVID) resulting from SARS-CoV2 infection).

93. Formula (II): 【Chemical Formula 15】 (wherein R 2 and n are as defined in claim 1, R 1a is C(O)OR 5 and R 3a is C(O)OR 16 wherein R 5 and R 16 are as defined in claim 1); or Formula (IP2): 【Chemical Formula 16】 (wherein R 2 and n are as defined in claim 1, R 3 is as defined for the above formula (II), R 4a is a protected NH 2 group, a protected OH group, or a protected C(O)NH 2C substituted with a radical 1-6 a compound that is alkyl).

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