Drug compounds

Abstract

This invention relates to formula (1) X 1 -L-X 2 The compound or its salt or tautomer; wherein X 1 and X 2 The compounds are independently selected from formulas (2A), (2B), (2C), and (2D), wherein the connection point with group L is indicated, provided that at least one of X1 and X2 has formula (2A). The compounds enhance mitochondrial function, and therefore the present invention also provides pharmaceutical compositions containing the compounds and therapeutic uses of the compounds, particularly for diseases caused by or affected by mitochondrial dysfunction. (2A) (2B) (2C) (2D)

Description

Technical Field

[0001] This invention relates to compounds that enhance mitochondrial function, pharmaceutical compositions containing said compounds, and the therapeutic use of said compounds, particularly in the treatment of diseases caused by or affected by mitochondrial dysfunction. Background Technology

[0002] Mitochondria are complex intracellular organelles that play a crucial role in cellular homeostasis. Mitochondria, responsible for coordinating cellular energy production, are essential for sustaining life [Osellame 2012]. Chemical energy is produced by mitochondria through oxidative phosphorylation (OXPHOS) and stored as adenosine triphosphate (ATP). ATP is produced by mitochondria through a variety of chemical reactions collectively known as the Krebs cycle, or tricarboxylic acid (TCA) cycle. This complex set of cellular reactions is driven by the proton gradient across the inner mitochondrial membrane [Papa 2012]. In addition to supplying energy, mitochondria are involved in a variety of other processes, such as cell signaling, cell differentiation, and regulation of cell death [Finkel et al., Circulation Research. 2015;116:1810-1819; McBride 2006].

[0003] Oxidative phosphorylation (OXPHOS) is a metabolic pathway responsible for producing most of the cell's energy. The human mitochondrial genome contains the genetic code for 13 proteins, all of which encode essential components of OXPHOS. The OXPHOS system consists of five multi-protein complexes, individual subunits of which are encoded by the mitochondrial or nuclear genome [Smeitink 2001]. This complex series of biochemical reactions converts the transmembrane electrochemical proton gradient energy into mechanical energy via ATP synthase, ultimately catalyzing the chemical bond between ADP and phosphate (P) to form ATP [Friedman 2014].

[0004] Free radicals generated during ATP synthesis can cause DNA damage, making mitochondrial DNA more susceptible to damage than DNA in other cells. Mitochondria also lack some of the protective mechanisms responsible for preventing DNA damage found in other cellular systems. Furthermore, gene mutations can lead to reduced mitochondrial production. All these factors contribute to decreased mitochondrial activity. Mitochondrial abnormalities, characterized by decreased mitochondrial function and the accumulation of damaged mitochondria, have been observed in various cell types and tissues. Some diseases may be caused by mutations or deletions in the mitochondrial genome, while others may be caused by disorders of the mitochondrial respiratory system or other mitochondrial functional impairments [Wallace 1999]. In patients with mtDNA mutations, genetic and clinical manifestations are further complicated by the presence of multiple mtDNA genomes in individual cells, resulting in a mixture of mutant and wild-type genomes (heterogeneity) in the same cell or tissue [Stewart 2015].

[0005] Mitochondrial disorders are common genetic disorders of energy metabolism, affecting approximately 1 in 5,000 people. These disorders are typically incurable and exhibit genetic and clinical heterogeneity [DiMauro 2008]. This heterogeneous group of disorders can manifest as encephalopathy, myopathy, or multiple organ syndrome in children or adults. The disorders are characterized by insufficient activity of one or more mitochondrial respiratory chain (RC) complexes involved in OXPHOS [complexes I (CI) to V] and / or reduced levels of OXPHOS complex homeostasis, leading to decreased ATP production [Nsiah-Sefaa 2016]. The disorders are most commonly caused by genetic mutations in the mitochondrial or nuclear genome, exhibiting clinical heterogeneity and phenotypic diversity, with these mutations sharing OXPHOS deficiency and impaired ATP synthesis. Furthermore, the presence of mtDNA in all human tissues implies dysfunction in multiple organ systems. The most commonly affected organ systems are the nervous, muscular, cardiac, and endocrine systems. Regardless of phenotypic presentation, all mitochondrial disorders can be considered defects in energy metabolism and cellular function [Schaefer 2004].

[0006] Mitochondrial myopathy is a clinically heterogeneous disease that can affect multiple systems beyond skeletal muscle and is typically defined by morphological abnormalities of muscle mitochondria. These represent the largest subset of primary mitochondrial diseases, making them the most common congenital metabolic disorders [DiMauro 2008]. Mitochondrial myopathy associated with mitochondrial export dysfunction has been identified in several ways, including: mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episode (MELAS), one of the most common maternally inherited mitochondrial diseases [Lin 2014]; myoclonic epilepsy and ragged-red fibers (MERRF), in which 80-90% of patients carry mutations in the tRNALys gene in mtDNA, leading to inefficient ATP production [Wu 2010]; and mitochondrial DNA-associated Leigh syndrome and neurogenic muscle weakness, ataxia, and retinitis pigmentosa (NARP), which are part of a series of progressive neurodegenerative diseases caused by abnormal mitochondrial energy production [Thorburn].

[2017] ; Lechner syndrome, a devastating neurodegenerative disease with multiple pathogenic mutations in mitochondrial and nuclear genes encoding multiple components of OXPHOS [Baertling 2014]; Mitochondrial dysfunction in fatty acid oxidation disorders (such as long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD)) [Olpin 2005, Wajner 2016]; Maple syrup urine disease (MSUD), a congenital metabolic disorder that leads to the accumulation of DNA damage and corresponding mitochondrial dysfunction [Strand 2014]; Luft disease, clinically characterized by hypermetabolism caused by extensive uncoupling of mitochondrial respiration in skeletal muscle tissue [Luft 1994]; Chronic progressive external ophthalmoplegia (CPEO), also known as progressive external ophthalmoplegia (PEO), is a disease characterized by slow, progressive paralysis of the external ophthalmos [Luft 1994, Man

[2005] ; Charcot-Marie-Tooth disease type 2;CMT2, in which mutations in the mitochondrial fusion protein 2 gene (MFN2) may account for at least one-third of cases, and reduced oxidative phosphorylation efficiency in MFN2-associated CMT2 may contribute to the pathophysiology of axonal neuropathy [Züchner 2004, Loiseau 2007]; Leber hereditary optic neuropathy (LHON), a primary mtDNA disorder characterized by vision loss in young adults [Luft 1994]; and hereditary conditions such as Kearns-Sayre syndrome (KSS) [Pieczenik 2007], myoneurogenic gastrointestinal encephalopathy (MNGIE) [Pieczenik 2007], and Barth syndrome. (An X-linked cardiomyopathy caused by tafazzin mutations) [Gonzalvez 2013], all are associated with mitochondrial dysfunction. Furthermore, mitochondrial DNA depletion syndromes (MDS) are a group of genetically and clinically heterogeneous autosomal recessive disorders characterized by severely reduced mtDNA levels, leading to impaired energy production in affected tissues and organs; these disorders include TK2-related and POLG-related syndromes [El-Hattab 2013].

[0007] In addition, mitochondrial mutations are associated with mitochondrial dysfunction in the following areas: maternally inherited epilepsy / mitochondrial tubulointerstitial kidney disease (MITKD) [Connor 2017]; mitochondrial deafness (DEAF) [Kototas 2007]; ataxia, myoclonus, and deafness (AMDF) [Park 2014]; hypertrophic cardiomyopathy (HCM) [Lucas 2003]; diabetes mellitus and deafness (DMDF) [Hutchin 200]; maternally inherited diabetes mellitus and deafness (MIDD) [Tsang 2018]; mitochondrial syndrome-related sensorineural hearing loss (SNHL) [Forli 2007]; focal segmental glomerulosclerosis (FSGS) associated with mitochondrial disease [Lim 2017]; autism spectrum disorder (ASD)

[2012] ; and progressive encephalopathy (PEM) [Kollberg].

[2006] ; Bilateral striatal necrosis (BSN) [Solano 2003]; Leber's hereditary optic neuropathy and dystonia (LDYT)

[2021] ; Maternally inherited cardiomyopathy (MICM) [Casali 1999]; Motor neuron disease (MND)

[2020] ; Myoclonic epilepsy / spasmodic epilepsy [Lamperti 2016]; Mitochondrial myopathy, lactic acidosis and sideroblastic anemia (MLASA)

[2014] ; Familial bilateral striatal necrosis (FBSN) [Thyagarajan 1995]; Epilepsy, stroke, optic atrophy and cognitive decline (Epilepsy, stroke, optatrophy, and cognitive decline; ESOC)

[2001] .

[0008] Mitochondrial dysfunction is not only a hallmark of rare, inherited mitochondrial disorders, but it is also associated with age-related diseases, including neurodegenerative diseases such as Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis (ALS), and Alzheimer's disease [Johri 2021]; metabolic disorders such as type 2 diabetes [Lowell 2005]; steatosis and non-alcoholic steatohepatitis (NASH) [Begriche 2006]; and obesity [Bournat 2010]. Furthermore, heart disease (such as ischemia-reperfusion, ischemic preconditioning, cardiomyopathy, and heart failure) [Lesnefsky 2001], as well as muscular dystrophy [Ryu 2016] and aging [Cui 2012] are all associated with mitochondrial dysfunction.

[0009] Cellular energy deficiency caused by mitochondrial dysfunction is a hallmark of mitochondrial diseases. This increases the likelihood of pharmacologically increasing cellular mitochondrial content to boost mitochondrial ATP production, as occurs in motor responses [Komen 2014]. A potential therapeutic approach to overcome impaired mitochondrial biogenesis is to effectively promote mitochondrial biogenesis through pharmacological means to compensate for OXPHOS deficiencies associated with mitochondrial respiratory dysfunction, thereby enriching the wild-type mitochondrial population [Wenz 2010, Moraes 2009]. Therefore, strategies aimed at improving mitochondrial function may provide an effective therapy for patients with mitochondrial diseases. It has been demonstrated that pharmacological stimulation of several proteins can improve mitochondrial biogenesis and function. For example, pharmacological activation of the transcriptional coactivator peroxisome proliferator-activated receptor gamma coactivator 1-α (PGC-1α) [Sandoval-Acuna 2014] has been shown; agonist activation of a combination of peroxisome proliferator-activated receptor (PPAR) and deoxyribonucleoside (dN) has been shown to increase mtDNA copy number and mitochondrial mass, thereby improving mitochondrial respiratory function in a cellular MELAS model [Burgin 2020]; and AMPK-activating compounds have been shown to promote mitochondrial function by increasing the expression of genes involved in mitochondrial biogenesis [Herzig 2018]. Compounds such as nicotinamide nucleoside have been shown to increase mitochondrial biogenesis and systemic metabolism [Kahn 2014], and nicotinamide mononucleotide has been shown to improve age-related phenotypes in mice, including changes in energy metabolism and increased physical activity, by increasing NAD+ levels [Mills 2016]. Furthermore, compounds that enhance NAD (such as PARP inhibitors [Felici 2014] and CD38 inhibitors [Escande 2013], as well as compounds that interact with NAD(P)H:quinone oxidoreductase 1 (NQO1)) have been shown to improve mitochondrial quality and OXPHOS in cellular models of mitochondrial dysfunction [Seo, Kang-Sik 2018]. These findings in cellular and mouse models with different mitochondrial defects support strategies to improve diseases characterized by abnormal mitochondrial biology by increasing mitochondrial quality.

[0010] Therefore, it would be beneficial to develop compounds that can increase cellular mitochondrial activity, for example, by increasing mitochondrial mass. Summary of the Invention

[0011] This invention provides compounds selected from compounds that can be used to improve mitochondrial activity and / or increase mitochondrial mass.

[0012] Therefore, in the first embodiment of the present invention (Embodiment 1.1), a compound of formula (1) is provided: X 1 -LX 2 (1) or its salts or tautomers; wherein X 1 and X 2 Choose independently from equations (2A), (2B), (2C), and (2D): (2A) (2B) (2C) (2D) in Indicates the connection point with group L, provided that X 1 and X 2 At least one of them has equation (2A); R 2 Selected from hydrogen, C, optionally substituted with hydroxyl groups 1-4 The hydrocarbon group and a 5-6 membered cyclic aromatic group or a 3-6 membered cyclic non-aromatic group, wherein the cyclic aromatic group and the cyclic non-aromatic group are optionally substituted by one or more substituents selected from: halogen, C 1-4 Alkyl, hydroxyl, C 1-4 Alkoxy and C 1-4 Alkyl group; and R 3 and R 4 One of them is selected from hydrogen and C, which is optionally substituted with a 5-6 membered cyclic aromatic group. 1-4 The hydrocarbon group, wherein the cyclic aromatic group is optionally substituted by one or more substituents selected from: halogen, cyano, hydroxyl, C 1-4 Alkyl and C 1-4 Alkyl groups; and R 3 and R 4 The other one is hydrogen; L is an optionally substituted cyclic or acyclic linker group, which in X 1 With X 2 The chain length is 2 to 8 carbon atoms, wherein one of the carbon atoms may optionally be replaced by a heteroatom selected from O, N, S and their oxidized forms, provided that the heteroatom (if present) is in relation to X. 1 and X 2 There are always at least two carbon atoms between each of them.

[0013] The particularly preferred compound of formula (1) is defined in the following embodiments 1.2 to 1.95.

[0014] 1.2 The compound according to embodiment 1.1, wherein X 1 and X 2 same.

[0015] 1.3 The compound according to embodiment 1.1, wherein X 1 and X 2 different.

[0016] 1.4 The compound according to embodiment 1.1, wherein X 1 It has formula (2A).

[0017] 1.5 The compound according to embodiment 1.4, wherein X 2 It has formula (2A).

[0018] 1.6 The compound according to embodiment 1.4, wherein X 2 It has formula (2B).

[0019] 1.7 The compound according to embodiment 1.4, wherein X 2 It has formula (2C).

[0020] 1.8 The compound according to embodiment 1.4, wherein X 2 It has the formula (2D).

[0021] 1.9 The compound according to any one of embodiments 1.1 to 1.3, wherein the chain length of L is 2 to 6 carbon atoms, wherein one of said carbon atoms may optionally be replaced by a heteroatom selected from O, N, S, S(O) and SO2, provided that the heteroatom (if present) is in contact with X. 1 and X 2 There are always at least two carbon atoms between each of them.

[0022] 1.10 The compound according to embodiment 1.9, wherein the chain length of L is 2 to 6 carbon atoms, wherein one of the carbon atoms may optionally be replaced by a heteroatom selected from O, N and S, provided that the heteroatom (if present) is in contact with X. 1 and X 2 There are always at least two carbon atoms between each of them.

[0023] 1.11 The compound according to embodiment 1.9, wherein the chain length of L is 2 to 5 carbon atoms, wherein one of said carbon atoms may optionally be replaced by a heteroatom selected from O, N, S, S(O) and SO2, provided that the heteroatom (if present) is in contact with X. 1 and X 2 There are always at least two carbon atoms between each of them.

[0024] 1.12 The compound according to embodiment 1.11, wherein the chain length of L is 2 to 5 carbon atoms, wherein one of the carbon atoms may optionally be replaced by a heteroatom selected from O, N and S, provided that the heteroatom (if present) is in contact with X. 1 and X 2 There are always at least two carbon atoms between each of them.

[0025] 1.13 The compound according to any one of embodiments 1.1 to 1.3, wherein the chain length of L is 2 to 6 carbon atoms, wherein none of the carbon atoms are replaced by heteroatoms.

[0026] 1.14 The compound according to any one of embodiments 1.13, wherein the chain length of L is 2 to 5 carbon atoms, wherein none of the carbon atoms are replaced by heteroatoms.

[0027] 1.15 The compound according to any one of embodiments 1.1 to 1.13, wherein L has formula A 1 -BA 2 ,in: A 1 and A 2 C-type bonds independently selected from the bond and optionally substituted with one or more hydroxyl groups or halogens. 1-4 Hydrocarbon linkers; B is selected from the bond, C(R) b )2、N(R b ), O, S, S(O), SO2, 3 to 6-membered cyclic non-aromatic groups and 5 or 6-membered cyclic aromatic groups, wherein the cyclic non-aromatic groups and the cyclic aromatic groups are optionally surrounded by one or more groups R b Replace; and R b Selected from hydrogen, halogen, hydroxyl, C 1-6 hydrocarbon groups and -OC 1-6 Hydrocarbon groups, wherein each hydrocarbon group may optionally be substituted with one or more substituents selected from hydroxyl and halogen.

[0028] 1.16 The compound according to any one of embodiments 1.1 to 1.15, wherein A 1 and A 2 same.

[0029] 1.17 The compound according to any one of embodiments 1.1 and 1.3 to 1.16, wherein A 1 and A 2 different.

[0030] 1.18 The compound according to any one of embodiments 1.1 to 1.17, wherein A 1 For key.

[0031] 1.19 The compound according to any one of embodiments 1.1 to 1.18, wherein A 2 For key.

[0032] 1.20 The compound according to any one of embodiments 1.15 to 1.17, wherein A 1 and A 2 Independently selected from C that is optionally substituted with one or more hydroxyl groups or halogens. 1-4 Hydrocarbon linkers (e.g., C) 1-4 (Saturated hydrocarbon linker).

[0033] 1.21 The compound according to any one of embodiments 1.15 to 1.17, wherein A 1 and A 2 Independently selected from C that is optionally substituted with one or more hydroxyl groups or halogens. 1-3 Hydrocarbon linkers (e.g., C) 1-3 (Saturated hydrocarbon linker).

[0034] 1.22 The compound according to any one of embodiments 1.15 to 1.17, wherein A 1 and A 2 Independently selected from C14 groups that are optionally substituted with a hydroxyl or halogen substituent. 1-4 Hydrocarbon linkers (e.g., C) 1-4 (Saturated hydrocarbon linker).

[0035] 1.23 The compound according to any one of embodiments 1.15 to 1.17, wherein A 1 and A 2 Selected independently from C 1-3 Hydrocarbon linkers (e.g., C) 1-3 (Saturated hydrocarbon linker).

[0036] 1.24 The compound according to any one of embodiments 1.15 to 1.23, wherein A 1 and A 2 It is independently selected from methylene or ethylene.

[0037] 1.25 The compound according to any one of embodiments 1.15 to 1.24, wherein B is selected from the bond, C(R) b )2、N(R b ), O, S, S(O), SO2, 3 to 6-membered cyclic non-aromatic groups and 5 or 6-membered cyclic aromatic groups, wherein the cyclic non-aromatic groups and the cyclic aromatic groups are optionally surrounded by one or more groups R b replace.

[0038] 1.26 The compound according to any one of embodiments 1.15 to 1.25, wherein B is selected from bonds, O, SO2, and optionally surrounded by one or more groups R. b Substituted 3- to 6-membered cyclic non-aromatic groups.

[0039] 1.27 The compound according to any one of embodiments 1.15 to 1.26, wherein B is selected from bonds, O, SO2, and optionally surrounded by one or more groups R. b Substituted 3 to 6-membered carbon ring non-aromatic groups.

[0040] 1.28 The compound according to any one of embodiments 1.15 to 1.24, wherein B is a bond.

[0041] 1.29 The compound according to any one of embodiments 1.15 to 1.24, wherein C(R) b )2 is the key.

[0042] 1.30 The compound according to any one of embodiments 1.15 to 1.24, wherein B is N(R b ).

[0043] 1.31 The compound according to any one of embodiments 1.15 to 1.24, wherein B is O.

[0044] 1.32 The compound according to any one of embodiments 1.15 to 1.24, wherein B is S.

[0045] 1.33 The compound according to any one of embodiments 1.15 to 1.24, wherein B is S(O).

[0046] 1.34 The compound according to any one of embodiments 1.15 to 1.24, wherein B is SO2.

[0047] 1.35 The compound according to any one of embodiments 1.15 to 1.24, wherein B is a 3- to 6-membered cyclic non-aromatic group and a 5- or 6-membered cyclic aromatic group, wherein the cyclic non-aromatic group and the cyclic aromatic group are optionally surrounded by one or more groups R. b replace.

[0048] 1.36 The compound according to embodiment 1.35, wherein B is optionally coated with one or more groups R b Substituted 3 to 6-membered carbon rings or heterocyclic non-aromatic groups.

[0049] 1.37 The compound according to embodiment 1.36, wherein B is optionally coated with one or more groups R. b Substituted 3 to 6-membered carbon ring non-aromatic groups.

[0050] 1.38 The compound according to embodiment 1.36, wherein B is optionally coated with one or more groups R. b Substituted 4- to 6-membered carbon ring non-aromatic groups.

[0051] 1.39 The compound according to embodiment 1.38, wherein the non-aromatic carbocyclic group is selected from cyclohexyl, cyclopentyl, and cyclobutyl.

[0052] 1.40 The compound according to embodiment 1.39, wherein the non-aromatic group of the carbocyclic ring is cyclohexyl.

[0053] 1.41 The compound according to embodiment 1.40, wherein L has the following formula (3): (3) in Instructions and X 1 or X 2 The connection point of any of them, provided that there exists a connection point with X in equation (3). 1 A connection point and a connection with X 2 The connection point.

[0054] 1.42 The compound according to embodiment 1.39, wherein the non-aromatic group of the carbocyclic ring is cyclopentyl.

[0055] 1.43 The compound according to embodiment 1.39, wherein the non-aromatic group of the carbocyclic ring is cyclobutyl.

[0056] 1.44 The compound according to any one of embodiments 1.15 to 1.43, wherein R b Selected from hydrogen, halogen, hydroxyl, C 1-4 hydrocarbon groups and -OC 1-4 Hydrocarbon groups, wherein each hydrocarbon group may be saturated and may optionally be substituted by one or more substituents selected from hydroxyl, halogen and cyano groups.

[0057] 1.45 The compound according to any one of embodiments 1.15 to 1.44, wherein R b Selected from hydrogen, halogen, hydroxyl, C 1-2 hydrocarbon groups and -OC 1-2 Hydrocarbon groups, wherein each hydrocarbon group may be saturated and may optionally be substituted by one or more substituents selected from hydroxyl and halogen.

[0058] 1.46 The compound according to any one of embodiments 1.15 to 1.44, wherein R b Selected from hydrogen, fluorine, hydroxyl, C 1-2 hydrocarbon groups and -OC 1-2 Hydrocarbon group.

[0059] 1.47 The compound according to embodiment 1.46, wherein R b It is hydrogen.

[0060] 1.48 The compound according to embodiment 1.46, wherein R b It is fluorine.

[0061] 1.49 The compound according to any one of embodiments 1.1 to 1.48, wherein L is C 2-6 Hydrocarbon linkers (e.g., those with the formula (CH2)) n (where n is 2 to 6).

[0062] 1.50 The compound according to any one of embodiments 1.1 to 1.49, wherein L is C 2-5 Hydrocarbon linkers (e.g., those with the formula (CH2)) n (where n is 2 to 5).

[0063] 1.51 The compound according to any one of embodiments 1.1 to 1.50, wherein L is C 2-4 Hydrocarbon linkers (e.g., those with the formula (CH2)) n (where n is 2 to 4).

[0064] 1.52 The compound according to any one of embodiments 1.1 to 1.51, wherein L is C 2-3 Hydrocarbon linkers (e.g., those with the formula (CH2)) n (where n is 2 to 3).

[0065] 1.53 The compound according to any one of embodiments 1.49 to 1.52, wherein the hydrocarbon linker is a saturated hydrocarbon linker.

[0066] 1.54 The compound according to any one of embodiments 1.1 to 1.52, wherein L contains up to 15 non-hydrogen atoms.

[0067] 1.55 The compound according to any one of embodiments 1.1 to 1.54, wherein L contains up to 10 non-hydrogen atoms.

[0068] 1.56 The compound according to any one of embodiments 1.1 to 1.55, wherein L contains up to 8 non-hydrogen atoms.

[0069] 1.57 The compound according to any one of embodiments 1.1 to 1.56, wherein L contains up to 6 non-hydrogen atoms.

[0070] 1.58 The compound according to any one of embodiments 1.1 to 1.57, wherein L contains up to 5 non-hydrogen atoms.

[0071] 1.59 The compound according to any one of embodiments 1.1 to 1.58, wherein L contains up to 4 non-hydrogen atoms.

[0072] 1.60 The compound according to any one of embodiments 1.1 to 1.59, wherein L contains up to 3 non-hydrogen atoms.

[0073] 1.61 The compound according to any one of embodiments 1.1 to 1.60, wherein L contains at most three atoms other than carbon and hydrogen.

[0074] 1.62 The compound according to any one of embodiments 1.1 to 1.61, wherein L contains 0, 1 or 2 atoms other than carbon and hydrogen.

[0075] 1.63 The compound according to any one of embodiments 1.1 to 1.62, wherein L contains 0 or 1 atom other than carbon and hydrogen.

[0076] 1.64 The compound according to any one of embodiments 1.1 to 1.63, wherein L contains 0 atoms other than carbon and hydrogen.

[0077] 1.65 The compound according to any one of embodiments 1.1 to 1.64, wherein L contains one atom other than carbon and hydrogen.

[0078] 1.66 The compound according to any one of embodiments 1.1 to 1.65, wherein L contains two atoms other than carbon and hydrogen.

[0079] 1.67 The compound according to any one of embodiments 1.1 to 1.66, wherein R 2 Selected from hydrogen, C 1-4 Saturated hydrocarbon group and 5-6 membered cyclic aromatic group optionally substituted with one or more substituents selected from the following: halogen, C 1-4 Alkyl, hydroxyl, C 1-4 Alkoxy and C 1-4 Alkyl group.

[0080] 1.68 The compound according to any one of embodiments 1.1 to 1.67, wherein R 2 For hydrogen, C 1-4 Alkyl groups and phenyl groups optionally substituted with one or more substituents selected from the following: halogens, C 1-4 Alkyl, hydroxyl, C 1-4 Alkoxy and C 1-4 Alkyl group.

[0081] 1.69 The compound according to any one of embodiments 1.1 to 1.67, wherein R 2 Selected from hydrogen, C 1-4 Alkyl groups and 5-6 membered cyclic aromatic groups.

[0082] 1.70 The compound according to any one of embodiments 1.1 to 1.69, wherein R 2 Selected from hydrogen, C 1-4 Alkyl groups and unsubstituted phenyl groups.

[0083] 1.71 The compound according to any one of embodiments 1.1 to 1.70, wherein R 2 It is selected from hydrogen, methyl, ethyl and phenyl.

[0084] 1.72 The compound according to any one of embodiments 1.1 to 1.70, wherein R 2 Selected from hydrogen and C 1-4 alkyl.

[0085] 1.73 The compound according to any one of embodiments 1.1 to 1.70, wherein R 2 Selected from hydrogen and C 1-3 alkyl.

[0086] 1.74 The compound according to any one of embodiments 1.1 to 1.71, wherein R 2 Selected from hydrogen, methyl, and ethyl.

[0087] 1.75 The compound according to any one of embodiments 1.1 to 1.74, wherein R 2 It is hydrogen.

[0088] 1.76 The compound according to any one of embodiments 1.1 to 1.74, wherein R 2 It is a methyl group.

[0089] 1.77 The compound according to any one of embodiments 1.1 to 1.74, wherein R 2 It is a phenyl group.

[0090] 1.78 The compound according to any one of embodiments 1.1 to 1.77, wherein R 4 It is hydrogen.

[0091] 1.79 The compound according to any one of embodiments 1.1 to 1.78, wherein R 3 Selected from hydrogen and C, optionally substituted with 5-6 membered cyclic aromatic groups. 1-4 The hydrocarbon group, wherein the cyclic aromatic group is optionally substituted by one or more substituents selected from: halogen, cyano, hydroxyl, C1-4 Alkyl and C 1-4 Alkyl group.

[0092] 1.80 The compound according to any one of embodiments 1.1 to 1.79, wherein R 3 Selected from hydrogen and C, optionally substituted with 5-6 membered cyclic aromatic groups. 1-3 The hydrocarbon group, wherein the cyclic aromatic group is optionally substituted by one or more substituents selected from: halogen, cyano, hydroxyl, C 1-4 Alkyl and C 1-4 Alkyl group.

[0093] 1.81 The compound according to any one of embodiments 1.1 to 1.80, wherein R 3 Selected from hydrogen and C, optionally substituted with 5-6 membered cyclic aromatic groups. 1-3 The hydrocarbon group, wherein the cyclic aromatic group is optionally substituted by one or more substituents selected from the group consisting of cyano, C, ... 1-2 Alkyl and C 1-2 Alkyl group.

[0094] 1.82 The compound according to any one of embodiments 1.1 to 1.81, wherein R 3 Selected from hydrogen and C groups optionally substituted with phenyl or pyridyl groups 1-3 The alkyl group, wherein the phenyl or pyridyl group is optionally substituted with one or more substituents selected from cyano and methoxy.

[0095] 1.83 The compound according to any one of embodiments 1.1 to 1.82, wherein R 3 Selected from hydrogen and C groups optionally substituted with phenyl or pyridyl groups 1-3 The alkyl group, wherein the phenyl or pyridyl group is optionally substituted by one or more substituents selected from the following: cyano, C 1-2 Alkyl and C 1-2 Alkyl group.

[0096] 1.84 The compound according to any one of embodiments 1.1 to 1.83, wherein R 3 Selected from hydrogen and C groups optionally substituted with phenyl or other groups 1-3 The alkyl group, wherein the phenyl group is optionally substituted by one or more substituents selected from the group consisting of cyano, C, ... 1-2 Alkyl and C 1-2 Alkyl group.

[0097] 1.85 The compound according to any one of embodiments 1.1 to 1.84, wherein R 3 Selected from hydrogen and C244, optionally substituted with phenyl 1-3The phenyl group is a hydrocarbon group, wherein the phenyl group is optionally substituted with one or more substituents selected from cyano and methoxy.

[0098] 1.86 The compound according to any one of embodiments 1.1 to 1.79, wherein R 3 Selected from hydrogen and C 1-4 Hydrocarbon group.

[0099] 1.87 The compound according to embodiment 1.86, wherein R 3 Selected from hydrogen and noncyclic C 1-4 Hydrocarbon group.

[0100] 1.88 The compound according to embodiment 1.87, wherein R 3 Selected from hydrogen and noncyclic C 1-3 Hydrocarbon group.

[0101] 1.89 The compound according to any one of embodiments 1.79 to 1.88, wherein the hydrocarbon group is a saturated hydrocarbon group.

[0102] 1.90 The compound according to embodiment 1.89, wherein R 3 It is selected from hydrogen, methyl, ethyl, propyl, isopropyl and prop-2-enyl.

[0103] 1.91 The compound according to embodiment 1.90, wherein R 3 It is hydrogen.

[0104] 1.92 The compound according to any one of embodiments 1.1 to 1.91, wherein X 1 and / or X 2 It has the following formula (2A-i): (2A-i) Where R 2 R 3 and R 4 As defined in any one of Implementation Schemes 1.1 to 1.91.

[0105] 1.93 The compound according to any one of embodiments 1.1 to 1.91, wherein X 1 and / or X 2 It has the following formula (2A-ii): (2A-ii) Where R 2 R 3 and R 4 As defined in any of Implementation Schemes 1.1 to 1.91.

[0106] 1.94. The compound according to any one of embodiments 1.1 to 1.91, wherein X 1 and / or X 2 It has the following formula (2A-iii): (2A-iii) Where R 2 R 3 and R 4 As defined in any of Implementation Schemes 1.1 to 1.91.

[0107] 1.95. The compound according to any one of embodiments 1.1 to 1.91, wherein X 1 and / or X 2 It has the following formula (2A-iv): (2A-iv) Where R 2 R 3 and R 4 As defined in any of Implementation Schemes 1.1 to 1.91.

[0108] 1.96 The compound according to any one of embodiments 1.1 to 1.91, wherein X 1 Having the formula 2A-i and X 2 It has the formula 2A-i.

[0109] 1.97 The compound according to any one of embodiments 1.1 to 1.92, wherein X 1 Having Equation 2A-iii and X 2 It has Equation 2A-iii.

[0110] 1.98 The compound according to any one of embodiments 1.1 to 1.3, wherein X 1 and / or X 2 Groups AA to AI are selected from those in Table 1 below, where the asterisk marks the junction with L:

[0111] 1.99 The compound according to embodiment 1.98, wherein X 1 and / or X 2 It has the following formula (AE): (AE) in Indicates the connection point with group L.

[0112] 1.100 The compound according to any one of embodiments 1.1 to 1.3, wherein L is selected from groups BA to CC in Table 2 below, wherein the asterisk and X1 and X 2 Connection points:

[0113] 1.101 A compound selected from the title compounds of Examples 1 to 56 herein.

[0114] 1.102 A compound having the following structural formula (1A): (1A).

[0115] 1.103 A compound having the following structural formula (1B): (1B).

[0116] 1.104 The compound according to any one of embodiments 1.1 to 1.103, which is in salt form.

[0117] 1.105 The compound according to embodiment 1.104, wherein the salt is an acid addition salt.

[0118] 1.106 The compound according to embodiment 1.104 or embodiment 1.105, wherein the salt is a pharmaceutically acceptable salt.

[0119] 1.107 The compound according to any one of embodiments 1.1 to 1.103 is in a non-salt (e.g., free base) form.

[0120] 1.108 The compound according to any one of embodiments 1.1 to 1.107, which is in solvate form.

[0121] 1.109 The compound according to embodiment 1.108, wherein the solvate is a hydrate.

[0122] definition Unless the context otherwise indicates, references to “carbocyclic” and “heterocyclic” groups as used herein shall include both aromatic and non-aromatic ring systems. Thus, by way of example, the term “carbocyclic and heterocyclic groups” in its scope includes aromatic, non-aromatic, unsaturated, partially saturated, and fully saturated carbocyclic and heterocyclic systems.

[0123] The carbocyclic or heterocyclic group may be aryl or heteroaryl. Aryl or heteroaryl can be monocyclic or bicyclic as defined herein. As used herein, the term "aryl" refers to a carbocyclic group with aromatic characteristics, and the term "heteroaryl" is used herein to refer to a heterocyclic group with aromatic characteristics. Where the context permits, the terms "aryl" and "heteroaryl" may cover bicyclic systems in which both rings are aromatic, or one ring is non-aromatic and the other is aromatic. In such bicyclic systems containing one aromatic and one non-aromatic group, the group may be linked by an aromatic ring or by a non-aromatic ring.

[0124] The term "non-aromatic group" refers to unsaturated ring systems, partially saturated, and fully saturated carbocyclic and heterocyclic systems that do not possess aromatic characteristics. The terms "unsaturated" and "partially saturated" refer to ring structures containing atoms sharing more than one valence bond; for example, the ring contains at least one multiple bond, such as a C=C=C bond. The term "saturated" refers to a ring structure where no multiple bonds exist between the ring atoms. Saturated carbocyclic groups include cycloalkyl groups: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. Partially saturated carbocyclic groups include cycloalkenyl groups: cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl. Non-aromatic heterocyclic groups include aziridine, pyrrolidine, piperidine, aziridine, piperazine, morpholine, thiomorpholine, thiomorpholine S-oxide and S,S-dioxide, pyran (2H-pyran or 4H-pyran), dihydrothiophene, dihydropyran, dihydrofuran, dihydrothiazole, tetrahydrofuran, tetrahydrothiophene, dioxane, tetrahydropyran, tetrahydrothiopyran, S-oxide, SS-dioxide, imidazoline, imidazolidinone, oxazoline, thiazoline, pyrazoline, and pyrazolidine.

[0125] Unless otherwise stated, the term "hydrocarbyl" as used herein refers to an aliphatic, alicyclic, aromatic, and acyclic group having an all-carbon backbone and consisting of carbon and hydrogen atoms. Examples of hydrocarbyl groups include alkyl, cycloalkyl, cycloalkenyl, carbocyclic aryl, alkenyl, ynyl, cycloalkylalkyl, cycloalkenylalkyl, and carbocyclic aralkyl, arylenyl, and aryynyl. Such groups may be unsubstituted or, where specified, substituted with one or more substituents as defined herein. In some cases, as defined herein, one or more, but not all, of the carbon atoms in a hydrocarbyl group may be substituted with another atom or group of atoms. Hydrocarbyl groups may be saturated or unsaturated.

[0126] In equation (1), L is the distance from X. 1 With X 2 The linking groups between them have a chain length of 2 to 8 carbon atoms.

[0127] As used in this article, the term "chain length" refers to the distance between two specified points (e.g., X). 1 With X 2 The number of atoms (e.g., carbon atoms) extending from the line between the atoms in the chain, excluding any atoms branching off from the chain. Thus, for example, the chain length of the group -(CH2)3-C(CH3)2-CH2- is 5 carbon atoms.

[0128] When the chain includes a ring structure, the chain length is based on the minimum number of carbon atoms (or other atoms) required to traverse the ring. For example, in the following linking groups: The chain length consists of two carbon atoms from the methylene group and four intermediate carbon atoms from the cyclohexane ring, resulting in a total chain length of 6 carbon atoms.

[0129] For some of the compounds disclosed in this paper, X was shown. 1 Connection point with L and X 2 Stereochemistry at the junction with L. For the other compounds disclosed herein, X is not shown. 1 Connection point with L and X 2 Stereochemistry at the junction with L. It should be understood that if the stereochemistry at the junction is not disclosed, it may take any form.

[0130] When using the term C x-y When using hydrocarbon linkers, the integers x and y refer to the number of carbon atoms in the hydrocarbon linker (not the chain length of the linker).

[0131] The claims of this application should be understood to cover only stable compounds. The claims are not intended to cover any combination of groups that would produce unstable compounds. For example, if the definition of protection covers a compound having OO or ON bonds, it should be understood that this compound is not intended to be protected because it is not a stable compound.

[0132] Salt The compounds of the present invention as defined in embodiments 1.1 to 1.109 may be presented in the form of salts.

[0133] The salts mentioned above (and also defined in implementation schemes 1.104, 1.105 and 1.106) are generally acid addition salts.

[0134] It can be done through conventional chemical methods, such as Pharmaceutical Salts: Properties, Selection, and UseP. Heinrich Stahl (ed.), Camille G. Wermuth (ed.), ISBN: 3-90639-026-8, Hardcover, 388 pages, August 2002. The method described herein is for the synthesis of salts from parent compounds. Generally, such salts can be prepared by reacting a compound in its free basic form with an acid in water or an organic solvent or a mixture thereof; generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are used.

[0135] Alternatively, a salt may be formed during the synthesis and subsequent separation of compound (1) or (2).

[0136] Acid addition salts (as defined in Embodiment 1.105) can be formed from a variety of acids (inorganic and organic acids). Examples of acid addition salts include salts formed from acids selected from the group consisting of: acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid (e.g., L-ascorbic acid), L-aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, butyric acid, (+)camphoric acid, camphorsulfonic acid, (+)-(1S)-camphor-10-sulfonic acid, decanoic acid, hexanoic acid, caprylic acid, cinnamic acid, citric acid, cyclohexane, dodecyl sulfate, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactopyric acid, gentian acid, glucoheponic acid, D-gluconic acid, glucuronic acid (e.g., D-glucuronic acid), glutamic acid (e.g., L-glutamic acid), α-oxo- Glutamic acid, glycolic acid, hippuric acid, hydrobromic acid, hydrochloric acid, hydroiodic acid, hydroxyethanesulfonic acid, (+)-L-lactic acid, (±)-DL-lactic acid, lactobionic acid, maleic acid, malic acid, (-)-L-malic acid, malonic acid, (±)-DL-mandelic acid, methanesulfonic acid, naphthalene-2-sulfonic acid, naphthalene-1,5-disulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, nitric acid, oleic acid, orotic acid, oxalic acid, palmitic acid, dihydroxynaphthoic acid, phosphoric acid, propionic acid, L-pyroglutamic acid, salicylic acid, 4-amino-salicylic acid, sebacic acid, stearic acid, succinic acid, sulfuric acid, tannic acid, (+)-L-tartaric acid, thiocyanate, p-toluenesulfonic acid, undecenoic acid and valeric acid, as well as acylated amino acids and cation exchange resins.

[0137] The salt forms of the compounds of this invention are generally pharmaceutically acceptable salts (Embodiment 1.106), and examples of pharmaceutically acceptable salts are discussed in Berge et al., 1977, “Pharmaceutically Acceptable Salts”. J. Pharm. Sci.,Volume 66, pp. 1-19. However, non-pharmaceutically acceptable salts can also be prepared as intermediates, which can then be converted into pharmaceutically acceptable salts. Such non-pharmaceutically acceptable salt forms, applicable for, for example, the purification or isolation of the compounds of the present invention, also form part of this invention.

[0138] The ability of a given compound of the present invention to form a stable acid addition salt will depend in part on the basicity of the free base form of the compound and the acidity of the acid. Preferably, although not necessarily required, there is a difference of at least two pKa units between the pKa of the acid and the pKa of the conjugate acid of the base. The acid may be selected from acids with a pKa of 3.5 or lower, for example, 3.0 or lower. Therefore, in another embodiment, the present invention provides: 1.110 The compound according to any one of embodiments 1.1 to 1.106, 1.108 and 1.109 is in the form of an acid addition salt formed with an acid having a pKa of 3.5 or lower (e.g. -7.0 to +3.5).

[0139] 1.111 The compound according to any one of embodiments 1.1 to 1.106, 1.108 and 1.109 is in the form of an acid addition salt formed with an acid having a pKa of 3.0 or lower.

[0140] 1.112 The compound according to embodiment 1.110 or embodiment 1.111, wherein the acid forming the acid addition salt is selected from hydrochloric acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, naphthalenesulfonic acid, malonic acid, maleic acid and fumaric acid.

[0141] 1.113 The compound according to embodiment 1.112, wherein the acid forming the acid addition salt is selected from hydrochloric acid, sulfuric acid and methanesulfonic acid.

[0142] Crystalline form The hydrochloride salts of the compounds of formula (1) may exist in amorphous or crystalline form, as described in Example 1 of this document, and it is assumed that other salt forms of the compounds will also exist in amorphous and crystalline forms.

[0143] Therefore, in another embodiment (embodiment 1.114), the present invention provides an acid addition salt of the compound of formula (1) which is substantially crystalline.

[0144] The term “substantially crystalline” refers to the form of a compound of formula (1) in which it is 50% to 100% crystalline. Within this range, a compound of formula (1) may be at least 55% crystalline, or at least 60% crystalline, or at least 70% crystalline, or at least 80% crystalline, or at least 90% crystalline, or at least 95% crystalline, or at least 98% crystalline, or at least 99% crystalline, or at least 99.5% crystalline, or at least 99.9% crystalline.

[0145] Therefore, in another embodiment (embodiments 1.115 to 1.118), the present invention provides: 1.115 An acid addition salt of a compound of formula (1) that is substantially crystalline, wherein it is at least 90% crystalline.

[0146] 1.116 An acid addition salt of a compound of formula (1) that is substantially crystalline, wherein it is at least 95% crystalline.

[0147] 1.117 An acid addition salt of a compound of formula (1) that is substantially crystalline, being at least 99% crystalline.

[0148] 1.118 An acid addition salt of a compound of formula (1) that is substantially crystalline, being at least 99.9% crystalline.

[0149] The compounds of the present invention may be in crystalline form either solvated (e.g., hydrated) or non-solvated (e.g., anhydrous).

[0150] As used herein, the term "anhydrous" does not exclude the possibility that some water may be present on or within the compound (e.g., a compound crystal). For example, some water may be present on the surface of the compound (e.g., a crystal) or a small amount of water may be present within the bulk of the compound (e.g., a crystal). Typically, the anhydrous form contains less than 0.4 molecules of water per molecule of the compound, and more preferably less than 0.1 molecules of water per molecule of the compound, such as 0 molecules of water.

[0151] When hydrated in its crystalline form, it may contain, for example, up to three water molecules of crystallization, and more typically up to two water molecules, such as one or two water molecules. It may also form non-stoichiometric hydrates, in which the number of water molecules present is less than one, or otherwise non-integer. For example, when less than one water molecule is present, each molecule of compound (1) may contain, for example, 0.4, or 0.5, or 0.6, or 0.7, or 0.8, or 0.9 water molecules.

[0152] Crystallization can be characterized using a variety of techniques, including X-ray powder diffraction (XRPD), single-crystal X-ray diffraction (see Example 1), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA). The properties of crystals under different humidity conditions can be analyzed using gravimetric vapor adsorption studies (such as dynamic vapor adsorption (DVS)).

[0153] The crystalline structure of a compound can be analyzed using X-ray powder diffraction (XRPD) solid-state technology. XRPD can be performed using conventional methods, such as those described herein (see examples below) and those in “Introduction to X-ray Powder Diffraction”, Ron Jenkins and Robert L. Snyder (John Wiley & Sons, New York, 1996). The presence of well-defined peaks in the XRPD diffraction pattern (relative to random background noise) indicates that the compound possesses a certain degree of crystallinity.

[0154] The X-ray powder pattern of the compound is characterized by the diffraction angle (2θ) and interplanar spacing (d) parameters of the X-ray diffraction pattern. These are related to Bragg's equation, nλ = 2d Sin θ, where n = 1; λ = wavelength of X-ray radiation; d = interplanar spacing; and θ = diffraction angle.

[0155] Alternatively, the crystalline structure of the salt form of compound (1) can be analyzed by single-crystal X-ray diffraction, as described in Example 1 below. The absolute stereochemical configuration of compound (1) has been determined using single-crystal X-ray diffraction studies.

[0156] Geometric isomers and tautomers The compounds of the present invention can exist in a variety of different geometric isomers and tautomers, and references to compounds of formulas (1) and (2) as defined in embodiments 1.1 to 1.118 include all such forms.

[0157] Optical isomers can be classified by their optical activity (i.e., they exist as + and - isomers or...). d and l The optical isomers can be characterized and identified by their isomer forms, or they can be characterized by their absolute stereochemistry using the "R and S" nomenclature developed by Cahn, Ingold, and Prelog, see [reference needed]. Advanced Organic Chemistry Jerry March, 6th edition, John Wiley & Sons, New Jersey, 2007, pp. 155-158; see also Cahn, Ingold, and Prelog. Angew. Chem. Int. Ed. Engl ., 1966, 5, 385-415.

[0158] Optical isomers can be separated by a variety of techniques, including chiral chromatography (via chiral support chromatography), and such techniques are well known to those skilled in the art.

[0159] As an alternative to chiral chromatography, optical isomers can be separated by forming diastereomeric salts with chiral acids such as (+)-tartaric acid, (-)-pyroglutamic acid, (-)-xylyl-L-tartaric acid, (+)-mandelic acid, (-)-malic acid and (-)-camphorsulfonic acid, or chiral amines such as (+)-1-phenylethylamine or (+)-1-(1-naphthyl)ethylamine, and by preferentially separating the diastereomeric isomers and subsequently dissociating the salt to give individual enantiomers of the free base.

[0160] In each of embodiments 1.1 to 1.118, the compounds of formula (1) and (2) are substantially optically pure; that is, they have an enantiomeric excess (ee) of at least 80% relative to any other optical isomer of the compounds of formula (1) and (2).

[0161] As used herein, the term “enantiomer excess” is used in its conventional sense, referring to the percentage excess of the enantiomer of interest (compound (1) or compound (2)). When the enantiomer excess is 80%, this corresponds to a composition containing 90% of the desired enantiomer + 10% of the other enantiomers = 100% (since 90% - 10% = 80%). More generally, compounds of formulas (1) and (2) have an optical purity of at least 82%, or at least 84%, or at least 86%, or at least 88%, or at least 90%, or at least 92%, or at least 94%, or at least 96%, or at least 98%, or at least 99%, or 100% (enantiomer excess).

[0162] Therefore, in another embodiment (embodiments 1.119 to 1.122), the present invention provides: 1.119 The compound according to any one of embodiments 1.1 to 1.118, wherein the compound of formula (1) or (2) has, as appropriate, the following optical purity: (i) at least 80%; or (ii) at least 82%; or (iii) at least 84%, or (iv) at least 86%; or (v) at least 88%; or (vi) at least 90%; or (vii) at least 92%; or (viii) At least 94%; or (ix) at least 96%; or (x) at least 98%; or (xi) at least 99%; or (xii) 100%.

[0163] 1.120 The compound according to embodiment 1.119, wherein the compound of formula (1) or (2) has an optical purity of at least 98%, depending on the circumstances.

[0164] 1.121 The compound according to embodiment 1.119, wherein the compound of formula (1) or (2) has an optical purity of at least 99%, as appropriate.

[0165] 1.122 The compound according to embodiment 1.119, wherein the compound of formula (1) or (2) has 100% optical purity as appropriate.

[0166] isotope In one embodiment, the compound of the invention as defined in any one of embodiments 1.1 to 1.122 is rich in deuterium at one or more sites.

[0167] Methods for deuterating organic compounds are known to those skilled in the art (see, for example, “Deuterium Discover and Applications in Organic Chemistry”, Jaemoon Yang, 2016, Elsevier, and “The Organic Chemistry of Isotopic Labelling”, James R Hanson, 2019, RSC Publishing).

[0168] When the compounds of embodiments 1.1 to 1.122 are deuterated, the percentage of deuterium atoms in the total hydrogen atoms of the compounds of the present invention is less than 2%, more typically less than 1%, more usually less than 0.1%, preferably less than 0.05%, and most preferably not more than 0.02%.

[0169] solvates The compounds defined in any of the embodiments 1.1 to 1.122 may be solvated or non-solventized.

[0170] Preferred solvates are those formed by incorporating a non-toxic, pharmaceutically acceptable solvent molecule (hereinafter referred to as a solvating solvent) into the solid structure (e.g., crystal structure) of the compound of the present invention. Examples of such solvents include water, alcohols (such as ethanol, isopropanol, and butanol), and dimethyl sulfoxide. Solvates can be prepared by recrystallizing the compound of the present invention with a solvent or a mixture of solvents containing a solvating solvent. Whether a solvate has been formed under any given condition can be determined by analyzing the crystals of the compound using well-known and standard techniques such as pyrolysis gravimetric analysis (TGE), differential scanning calorimetry (DSC), and X-ray crystallography.

[0171] Solvents can be stoichiometric or non-stoichiometric.

[0172] The particularly preferred solvates are hydrates, and examples of hydrates include hemihydrates, monohydrates and dihydrates.

[0173] For a more detailed discussion of solvates and methods for their preparation and characterization, see Bryn et al., Solid-State Chemistry of Drugs, 2nd ed., SSCI, Inc. of West Lafayette, IN, USA, 1999, ISBN 0-967-06710-3.

[0174] Complexes and cage-like inclusion compounds In one embodiment, the compound of any one of embodiments 1.1 to 1.122 is a complex of said compound (e.g., an inclusion complex or cage-like inclusion complex with a compound (such as cyclodextrin), or a complex with a metal).

[0175] Bioactivity The compounds as defined in any one of embodiments 1.1 to 1.122 can be used to enhance energy production in mitochondria. These compounds can be used to improve mitochondrial activity in vivo or in vitro.

[0176] The compound can be used to prevent (to reduce the likelihood of disease) or treat conditions or diseases involving components associated with mitochondrial dysfunction or abnormal activity.

[0177] For example, the compound may be suitable for the prevention or treatment of diseases and symptoms characterized by reduced mitochondrial activity. This reduced activity may be due to a decrease in the number of mitochondria produced by the body or due to reduced mitochondrial activity in the produced mitochondria.

[0178] As used herein, the term "treatment" generally refers to a beneficial therapeutic intervention. Such treatment can suppress symptoms caused by an underlying disease condition, even if the underlying disease condition persists; for example, the treatment can reduce or alleviate the symptoms of the disease, thereby making it easier for the patient to manage the symptoms.

[0179] As used in this article, the term "preventing" is generally used to refer to therapeutic interventions in which the onset of mitochondrial disease symptoms can be completely prevented or mitigated, even if the underlying cause of the disease (such as a genetic mutation) remains.

[0180] Therefore, in another embodiment (embodiments 2.1 to 2.21), the present invention provides: 2.1 The compound according to any one of embodiments 1.1 to 1.122, which is used in a medicament or therapy.

[0181] 2.2 The compound according to any one of embodiments 1.1 to 1.122, which is used for the prevention or treatment of mitochondrial diseases.

[0182] 2.3 The compound according to any one of embodiments 1.1 to 1.122, which is used for the prevention or treatment of diseases characterized by reduced mitochondrial activity.

[0183] 2.4 Use of a compound according to any one of embodiments 1.1 to 1.122 for the treatment or prevention of mitochondrial diseases.

[0184] 2.5 Use of a compound according to any one of embodiments 1.1 to 1.122 for the treatment or prevention of diseases characterized by reduced mitochondrial activity.

[0185] 2.6 A method for treating mitochondrial disease in an individual in need, the method comprising administering to the individual an effective amount of a compound according to any one of embodiments 1.1 to 1.122.

[0186] 2.7 A method for treating an individual in need of a disease characterized by reduced mitochondrial activity, the method comprising administering to the individual an effective amount of a compound according to any one of embodiments 1.1 to 1.122.

[0187] Mitochondrial diseases can manifest as mitochondrial myopathy. Nerve cells in the brain and muscles utilize large amounts of chemical energy; therefore, mitochondrial dysfunction can cause a variety of neuromuscular diseases.

[0188] Examples of such conditions and diseases include: Karns-Sell syndrome (KSS); Leigh syndrome; Maternally inherited Leigh syndrome (MILS); Mitochondrial DNA deletion syndrome (MDS); Mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS); Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE); Myosococcal epilepsy and broken red fibers (MERRF); Neuropathic ataxia and retinitis pigmentosa (NARP); Pearson syndrome or progressive extraocular muscle palsy (PEO); CPEO. Type 2 Chuck-Maley-Duss disease (CMT2); fatty acid oxidation disorder, i.e., long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) deficiency; maple syrup urine disease (MSUD); Left disease and Leber's hereditary optic neuropathy (LHON); maternally inherited epilepsy / mitochondrial tubulointerstitial nephropathy (MITKD), mitochondrial deafness (DEAF); ataxia, myoclonus, and deafness (AMDF); hypertrophic cardiomyopathy (HCM); diabetes mellitus and deafness (DMDF); maternally inherited diabetes mellitus and deafness (MIDD); mitochondrial syndrome-related sensorineural hearing loss. (SNHL); Focal segmental glomerulosclerosis associated with mitochondrial disease (FSGS); Autism spectrum disorder (ASD); Progressive encephalopathy (PEM); Bilateral striatal necrosis (BSN); Leber hereditary optic neuropathy and dystonia (LDYT); Maternally inherited cardiomyopathy (MICM); Motor neuron disease (MND); Myoclonic epilepsy / spasmodic epilepsy; Mitochondrial myopathy, lactic acidosis, and sideroblastic anemia (MLASA); Familial bilateral striatal necrosis (FBSN); Epilepsy, stroke, optic atrophy, and cognitive decline (ESOC).

[0189] Mitochondrial diseases can also be diabetes or deafness (DAD) and type 2 diabetes.

[0190] In addition to the diseases mentioned above, several other acquired conditions are believed to be related to mitochondrial dysfunction. These conditions include: ● Huntington's disease ● Cancer ● Alzheimer's disease Parkinson's disease ● Bipolar disorder ● Schizophrenia ● Aging and senescence ● Anxiety ● Cardiovascular diseases ● Sarcopenia ● Chronic fatigue syndrome Amyotrophic Lateral Sclerosis (ALS) ● Steatohepatitis and nonalcoholic steatohepatitis (NASH) ● Obesity ● Ischemia-reperfusion ● Ischemic preconditioning ● Cardiomyopathy ● Heart failure ● Muscular dystrophy.

[0191] Therefore, in another embodiment (embodiments 2.8 to 2.17), the present invention provides: 2.8 The compound according to any one of embodiments 1.1 to 1.122, which is used for the prevention or treatment of mitochondrial myopathy.

[0192] 2.9 Use of a compound according to any one of embodiments 1.1 to 1.122 for the treatment or prevention of mitochondrial myopathy.

[0193] 2.10 A method for treating mitochondrial myopathy in an individual in need, the method comprising administering to the individual an effective amount of a compound according to any one of embodiments 1.1 to 1.122.

[0194] 2.11 Use of the compound according to embodiment 2.8, the compound according to embodiment 2.9, or the method according to embodiment 2.10, wherein the mitochondrial myopathy is selected from: Karns-Sell syndrome (KSS); Reye's syndrome; maternally inherited Reye's syndrome (MILS); mitochondrial DNA deletion syndrome (MDS); mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS); mitochondrial neurogastrointestinal encephalomyopathy (MNGIE); myositis epilepsy and broken red fibers (MERRF); neuropathic ataxia and retinitis pigmentosa (NARP); Pearson syndrome or progressive extraocular muscle palsy (PEO); CPEO Type 2 Chuck-Maley-Duss disease (CMT2); fatty acid oxidation disorder, i.e., long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) deficiency; maple syrup urine disease (MSUD); Left disease and Leber's hereditary optic neuropathy (LHON); maternally inherited epilepsy / mitochondrial tubulointerstitial nephropathy (MITKD), mitochondrial deafness (DEAF); ataxia, myoclonus, and deafness (AMDF); hypertrophic cardiomyopathy (HCM); diabetes mellitus and deafness (DMDF); maternally inherited diabetes mellitus and deafness (MIDD); mitochondrial syndrome-related sensorineural hearing loss. (SNHL); Focal segmental glomerulosclerosis associated with mitochondrial disease (FSGS); Autism spectrum disorder (ASD); Progressive encephalopathy (PEM); Bilateral striatal necrosis (BSN); Leber hereditary optic neuropathy and dystonia (LDYT); Maternally inherited cardiomyopathy (MICM); Motor neuron disease (MND); Myoclonic epilepsy / spasmodic epilepsy; Mitochondrial myopathy, lactic acidosis, and sideroblastic anemia (MLASA); Familial bilateral striatal necrosis (FBSN); Epilepsy, stroke, optic atrophy, and cognitive decline (ESOC).

[0195] 2.12 The compound according to any one of embodiments 1.1 to 1.122, which is used for the prevention or treatment of diabetes or deafness (DAD) and type 2 diabetes.

[0196] 2.13 Use of a compound according to any one of embodiments 1.1 to 1.122 for the treatment or prevention of diabetes or deafness (DAD) and type 2 diabetes.

[0197] 2.14 A method for treating diabetes or deafness (DAD) and type 2 diabetes in an individual in need, the method comprising administering to the individual an effective amount of a compound according to any one of embodiments 1.1 to 1.122.

[0198] 2.15 The compound according to any one of embodiments 1.1 to 1.122, for the prevention or treatment of diseases selected from: Huntington's disease, cancer, Alzheimer's disease, Parkinson's disease, bipolar disorder, schizophrenia, aging and senility, anxiety disorders, cardiovascular disease, sarcopenia, chronic fatigue syndrome, amyotrophic lateral sclerosis, steatosis and non-alcoholic steatohepatitis (NASH), obesity, ischemia-reperfusion, ischemic preconditioning, cardiomyopathy, heart failure and muscular dystrophy.

[0199] 2.16 Use of a compound according to any one of embodiments 1.1 to 1.122 for the treatment or prevention of diseases selected from: Huntington's disease, cancer, Alzheimer's disease, Parkinson's disease, bipolar disorder, schizophrenia, aging and senility, anxiety disorders, cardiovascular diseases, sarcopenia, chronic fatigue syndrome, amyotrophic lateral sclerosis, steatosis and non-alcoholic steatohepatitis (NASH), obesity, ischemia-reperfusion, ischemic preconditioning, cardiomyopathy, heart failure and muscular dystrophy.

[0200] 2.17 A method for treating a disease in an individual in need, said disease being selected from Huntington's disease, cancer, Alzheimer's disease, Parkinson's disease, bipolar disorder, schizophrenia, aging and degeneration, anxiety disorders, cardiovascular disease, sarcopenia, chronic fatigue syndrome, amyotrophic lateral sclerosis, steatosis and nonalcoholic steatohepatitis (NASH), obesity, ischemia-reperfusion, ischemic preconditioning, cardiomyopathy, heart failure and muscular dystrophy, said method comprising administering to said individual an effective amount of a compound according to any one of embodiments 1.1 to 1.122.

[0201] 2.18 A method for treating an individual who has been diagnosed with and found to have a disease or symptom characterized by reduced mitochondrial activity, the method comprising administering to the individual an effective amount of a compound according to any one of embodiments 1.1 to 1.122.

[0202] 2.19 A method for diagnosing and treating a disease condition or symptom characterized by reduced mitochondrial activity, the method comprising: (i) screening a patient to determine whether the patient has or may have a disease or symptom that is susceptible to treatment with a compound that increases mitochondrial activity; and (ii) administering to the patient, if indicated that the patient is therefore susceptible to the disease or symptom, a compound according to any one of embodiments 1.1 to 1.122.

[0203] 2.20 Use of a compound according to any one of embodiments 1.1 to 1.122 for manufacturing a medicament for treating or preventing a disease condition or symptom in a patient who has been screened and identified as having or at risk of having a disease or symptom that is sensitive to treatment with a compound that increases mitochondrial activity.

[0204] 2.21 The compound according to any one of embodiments 1.1 to 1.122, used to treat or prevent a disease condition or symptom in a patient who has been screened and identified as having or at risk of having a disease or symptom that is sensitive to treatment with a compound that increases mitochondrial activity.

[0205] Diagnostic methods for determining whether a particular cancer is sensitive to treatment with the compounds of the present invention are described in the section entitled “Diagnostic Methods” below.

[0206] Determination of biological characteristics The ability of the compounds in implementation schemes 1.1 to 1.122 to increase mitochondrial activity can also be determined using the schemes described in the Examples section below.

[0207] Cell-based in vitro functional and phenotypic analyses can be used to simultaneously analyze mitochondrial parameters, thus providing a more comprehensive footprint of the effects of compounds on mitochondrial activity. Mitochondrial membrane potential (ΔψM) can be assessed using dyes that accumulate within mitochondria without affecting mitochondrial respiration. These dyes include, but are not limited to, acridine orange 10-nonyl bromide (NAO), MitoTracker™ Green FM, MITO-ID® Green, and MitoView™ Green. These dyes are widely used for imaging mitochondria to assess their intracellular localization and quantify mitochondrial abundance, thus providing indirect indicators of mitochondrial biogenesis and efficiency [Kitami 2012]. To assess mitochondrial biomass at the cellular level, reporter cell lines transfected with mitochondrial-targeting green fluorescent protein (GFP) can be used to quantify the expression of respiratory complex subunits (such as cytochrome c oxidase subunit IV (COXVIII)) in living cells [Wang 2012, Nilsson 2015]. Furthermore, the development of oxygen-dependent fluorescence quenching systems and high-resolution respirometry (HRR) has enabled the direct quantification of mitochondrial respiration in high-throughput formats. By combining multiple test results for all compounds, it is possible to define a comprehensive set of mitochondrial signatures, thereby enabling the clustering of novel chemical entities based on their impact on mitochondrial function [Andreux 2016].

[0208] Preferred compounds in embodiments 1.1 to 1.122 are compounds having 102% or greater, preferably 105% or greater, mitochondrial-size activity (measured using the methods described above, such as in the Mitotracker assay or the COX8 MTS reporter assay).

[0209] Therefore, in another embodiment (embodiments 2.22 to 2.26), the present invention provides: 2.22 The compound according to any one of embodiments 1.1 to 1.122 has 102% or greater mitochondrial-sized activity.

[0210] 2.23 The compound according to any one of embodiments 1.1 to 1.122 has 103% or greater mitochondrial-sized activity.

[0211] 2.24 The compound according to any one of embodiments 1.1 to 1.122 has 104% or greater mitochondrial-sized activity.

[0212] 2.25 The compound according to any one of embodiments 1.1 to 1.122 has 105% or greater mitochondrial-sized activity.

[0213] 2.26 The compound according to any one of embodiments 1.1 to 1.122, used in any one of embodiments 2.1 to 2.25 of the therapy, treatment, method or use.

[0214] Method for preparing the compounds of the present invention The present invention also provides a method for preparing the compound according to any one of embodiments 1.1 to 1.122.

[0215] The compound of formula (1) can be prepared by reacting a first carboxylic acid derivative of general formula (4) with an equivalent of a dihydroxy compound of general formula (5), optionally isolating the monosubstituted product (5A), and then reacting the compound of formula (5A) with an equivalent of a second carboxylic acid derivative of general formula (6): Where Y is defined as having one of the following structural formulas (7A), (7B), (7C), or (7D): (7A) (7B) (7C) (7D) in Indicates the connection point with the leaving group LG.

[0216] Therefore, in another embodiment (Embodiment 3.1), a method for preparing the compound according to any one of Embodiments 1.1 to 1.122 is provided, the method comprising: (a) Reacting an equivalent amount of a first carboxylic acid derivative of general formula (4) with an equivalent amount of a dihydroxy compound of general formula (5) to form a monosubstituted product having formula (5A): ,as well as (b) React the monosubstituted product (5) with an equivalent amount of a second carboxylic acid derivative of general formula (6); Wherein L is as defined in any one of embodiments 1.1 to 1.122, LG is a suitable leaving group, and Y is selected from (7A), (7B), (7C) or (7D): (7A) (7B) (7C) (7D) in Indicates the connection point with the leaving group LG.

[0217] The monosubstituted product can be separated after step (a) and before step (b).

[0218] Preferably, the reaction is carried out in an aprotic solvent at room temperature in the presence of a coupling agent. If the structures of the first and second carboxylic acid derivatives are different (i.e., if structures 4 and 6 are different), it is also preferred that a suitable protecting group be provided for one of the hydroxyl groups on the dihydroxy compound in step (a). Alternatively, if the structures of the first and second carboxylic acid derivatives are identical (i.e., if structures 4 and 6 are identical), the reaction can be carried out in a single step, wherein an equivalent of each of the first and second carboxylic acid derivatives (which are identical) reacts with an equivalent of the dihydroxy compound in a single step.

[0219] The following provides alternative implementation schemes (Schemes 3.2 to 3.13): 3.2 The method according to embodiment 3.1, wherein LG is selected from OH, halide ions, toluenesulfonate, methanesulfonate, benzenesulfonate, OR, or OC(O)R, where R is C 1-3 alkyl.

[0220] 3.3 The method according to implementation scheme 3.2 is selected from OH and Cl.

[0221] 3.4 The method according to implementation scheme 3.3, wherein LG is OH.

[0222] 3.4 The method according to any one of embodiments 3.1 to 3.4, wherein the reaction is carried out in an aprotic solvent.

[0223] 3.5 The method according to embodiment 3.3, wherein the aprotic solvent is tetrahydrofuran (THF) or dioxane.

[0224] 3.6 The method according to any one of embodiments 3.1 to 3.5, wherein the reaction is carried out in the presence of a nonnucleophilic base.

[0225] 3.7 The method according to embodiment 3.6, wherein the nonnucleophilic base is DMAP.

[0226] 3.8 The method according to any one of embodiments 3.1 to 3.7, wherein the reaction is carried out at room temperature.

[0227] 3.9 The method according to any one of embodiments 3.1 to 3.8, wherein the reaction is carried out in the presence of a coupling agent.

[0228] 3.10 The method according to embodiment 3.9, wherein the coupling agent is dicyclohexylcarbodiimide (DCC) or diisopropylcarbodiimide (DIC).

[0229] 3.11 The method according to any one of embodiments 3.1 to 3.10, wherein the compound of formula (4) is the same as the compound of formula (6).

[0230] 3.12 The method according to implementation scheme 3.11, wherein steps (a) and (b) are performed simultaneously.

[0231] 3.13 The method according to any one of embodiments 3.1 to 3.10, wherein the compound of formula (4) is different from the compound of formula (6).

[0232] 3.14 The method according to embodiment 3.13, wherein the method further comprises adding a protecting group to one of the hydroxyl groups on the dihydroxy compound prior to step (a), and removing the protecting group prior to step (b).

[0233] Compounds of formula (4) or (6) in which LG is OH (referred to herein as compounds of formula (8)) can be prepared by reacting a compound of formula (9) in which PG is a suitable nitrogen protecting group with an acid in a polar solvent to remove the N-Boc protecting group.

[0234] Therefore, in another embodiment (Embodiment 3.15), a method for preparing a compound of formula (4) or formula (6) as defined above is provided, wherein LG is OH, the method comprising: (a) Removal of protecting group PG from compound of formula (9): To form compounds of formula (8); The following provides an alternative implementation plan: 3.16 The method according to implementation scheme 3.15, wherein PG has the formula C(O)OR N , where R N C is arbitrarily replaced 1-4 Hydrocarbon group.

[0235] 3.17 The method according to implementation scheme 3.16, wherein R N It is tert-butyl.

[0236] 3.18 The method according to any one of embodiments 3.15 to 3.17, wherein step (a) is carried out in the presence of an acid.

[0237] 3.19 The method according to embodiment 3.18, wherein the acid is selected from trifluoroacetic acid and hydrochloric acid.

[0238] 3.20 The method according to any one of embodiments 3.15 to 3.19, wherein step (a) is carried out in a polar aprotic solvent.

[0239] 3.26 The method according to any one of embodiments 3.20 to 3.25, wherein step (a) is carried out in dichloromethane or diethyl ether.

[0240] Protective base In the methods described above, it may be necessary to protect one or more functional groups to prevent reactions from occurring at undesirable positions on the molecule. Examples of protecting groups and methods for protecting and deprotecting functional groups can be found in […]. Protective Groups in Organic Synthesis (P. Wuts; 5th edition; Wiley, 2014).

[0241] The hydroxyl group can be protected, for example, in the form of an ether (-OR) or an ester (-OC(=O)R), such as: tert-butyl ether; tetrahydropyranyl (THP) ether; anisole, benzhydryl / diphenylmethyl ether or triphenylmethyl ether; trimethylsilane ether or tert-butyldimethylsilane ether; or acetyl ester (-OC(=O)CH3, -OAc).

[0242] The aldehyde or ketone group can be protected, for example, in the form of an acetal (R-CH(OR)2) or a ketal (R2C(OR)2), wherein the carbonyl group (>C=O) is converted to a diether (>C(OR)2) by reaction with, for example, a primary alcohol. The aldehyde or ketone group is readily regenerated by hydrolysis with a large excess of water in the presence of an acid.

[0243] The amine group can be protected, for example, in the form of an amide (-NRCO-R) or a carbamate (-NRCO-OR), such as: methylamide (-NHCO-CH3); benzoxyamide (-NHCO-OCH2C6H5, -NH-Cbz or NH-Z); tert-butoxyamide (-NHCO-OC(CH3)3, -NH-Boc); 2-biphenyl-2-propoxyamide (-NHCO-OC(CH3)2C6H4C6H5, -NH-Bpoc); 9-fluorenylmethoxyamide (-NH-Fmoc); 6-nitroveratroloxyamide (-NH-Nvoc); 2-trimethylsilylethoxyamide (-NH-Teoc); 2,2,2-trichloroethoxyamide (-NH-Troc); allyloxyamide (-NH-Alloc); or 2(-phenylsulfonyl)ethoxyamide (-NH-Psec).

[0244] Other protecting groups of amines (such as cyclic amine groups and heterocyclic NH groups) include toluenesulphonyl / tosyl and methanesulphonyl / mesyl, benzyl (such as p-methoxybenzyl (PMB)) and tetrahydropyranyl (THP).

[0245] The carboxylic acid group can be protected in the form of an ester, for example: C 1-7 Alkyl esters (e.g., methyl esters; tert-butyl esters); C 1-7 Halogenated alkyl esters (e.g., C16) 1-7 Trihaloalkyl esters); Three C 1-7 Alkylsilyl-C 1-7 Alkyl ester; or C 5-20 Aryl-C 1-7 Alkyl esters (e.g., benzyl ester; nitrobenzene); or may be protected in the form of amides, such as methylamides. Thiol groups may be protected, for example in the form of thioethers (-SR), such as benzyl sulfide; acetamyl methyl ether (-S-CH2NHC(=O)CH3).

[0246] pharmaceutical preparations The compounds of the present invention are typically administered to patients in the form of pharmaceutical compositions. Therefore, in another embodiment of the invention (Embodiment 4.1), the present invention provides a pharmaceutical composition comprising a compound according to any one of Embodiments 1.1 to 1.122 and a pharmaceutically acceptable excipient.

[0247] In another implementation, the following is provided: 4.2 The pharmaceutical composition according to embodiment 4.1 comprises about 1% (w / w) to about 95% (w / w) of the compound according to any one of embodiments 1.1 to 1.122 and 99% (w / w) to 5% (w / w) of a pharmaceutically acceptable excipient or combination of excipients, and optionally one or more other therapeutically active ingredients.

[0248] 4.3 The pharmaceutical composition according to embodiment 4.2 comprises about 5% (w / w) to about 90% (w / w) of the compound according to any one of embodiments 1.1 to 1.122 and 95% (w / w) to 10% of a pharmaceutically acceptable excipient or combination of excipients, and optionally one or more other therapeutically active ingredients.

[0249] 4.4 The pharmaceutical composition according to embodiment 4.3 comprises about 10% (w / w) to about 90% (w / w) of the compound according to any one of embodiments 1.1 to 1.122 and 90% (w / w) to 10% of a pharmaceutically acceptable excipient or combination of excipients.

[0250] 4.5 The pharmaceutical composition according to embodiment 4.4 comprises about 20% (w / w) to about 90% (w / w) of the compound according to any one of embodiments 1.1 to 1.122 and 80% (w / w) to 10% of a pharmaceutically acceptable excipient or combination of excipients.

[0251] 4.6 The pharmaceutical composition according to embodiment 4.5 comprises about 25% (w / w) to about 80% (w / w) of the compound according to any one of embodiments 1.1 to 1.122 and 75% (w / w) to 20% of a pharmaceutically acceptable excipient or combination of excipients.

[0252] The pharmaceutical compositions of the present invention may be in any form suitable for oral, parenteral, topical, intranasal, intrabronchial, ocular, ocular, rectal, vaginal, or percutaneous administration. If the composition is intended for parenteral administration, it may be formulated for intravenous, intramuscular, intraperitoneal, or subcutaneous administration, or for direct delivery to the target organ or tissue by injection, infusion, or other means of delivery.

[0253] Drug dosage forms suitable for oral administration include tablets, capsules, pouches, pills, lozenges, syrups, solutions, sprays, powders, granules, elixirs and suspensions, sublingual tablets, sprays, powder tablets or patches and buccal patches.

[0254] Pharmaceutical compositions containing any one of the compounds according to embodiments 1.1 to 1.122 of the present invention (e.g., as defined in embodiment 4.1) can be formulated according to known techniques, see example Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA, USA.

[0255] Therefore, tablet compositions may contain a unit dose of the active compound and an inert diluent or carrier, such as sugars or sugar alcohols, such as lactose, sucrose, sorbitol, or mannitol; and / or non-sugar-derived diluents, such as sodium carbonate, calcium phosphate, talc, calcium carbonate, or cellulose or its derivatives, such as methylcellulose, ethylcellulose, hydroxypropyl methylcellulose, and starch, such as corn starch. Tablets may also contain such standard ingredients as binders and granulating agents (e.g., polyvinylpyrrolidone), disintegrants (e.g., expandable crosslinking polymers, such as crosslinked carboxymethyl cellulose), lubricants (e.g., stearates), preservatives (e.g., parabens), antioxidants (e.g., BHT), buffers (e.g., phosphate or citrate buffers), and foaming agents (e.g., citrate / bicarbonate mixtures). Such excipients are well known and need not be discussed in detail here.

[0256] Capsule formulations can be made from hard or soft gelatin and may contain active ingredients in solid, semi-solid, or liquid form. Gelatin capsules may be formed from animal gelatin or its synthetic or plant-derived equivalents.

[0257] Solid dosage forms (e.g., tablets, capsules, etc.) may be coated or uncoated, but are typically coated, such as with a protective film (e.g., wax or varnish) or a release-controlling coating. The coating (e.g., an Eudragit™ type polymer) can be programmed to release the active ingredient at a desired location within the gastrointestinal tract. Therefore, coatings can be selected to degrade under certain pH conditions within the gastrointestinal tract, thereby selectively releasing the compound in the stomach, ileum, or duodenum.

[0258] Alternatively, or in addition to coating, the drug product may be contained in a solid matrix comprising a release control agent, such as a release delay agent, which is adapted to selectively release the compound under varying acidity or alkalinity conditions in the gastrointestinal tract. Alternatively, the matrix material or release delay coating may be in the form of an erosive polymer (e.g., a maleic anhydride polymer) that is substantially continuously eroded as the dosage form passes through the gastrointestinal tract.

[0259] In one particular embodiment, the pharmaceutical composition of embodiment 4.1 is an enteric-coated solid dosage form, such as a coated tablet or capsule, wherein the coating is selected to be resistant to acidic conditions and degrade under pH conditions prevalent in the duodenum or ileum, thereby selectively releasing the compound in the duodenum or ileum.

[0260] Topical compositions include ointments, creams, sprays, patches, gels, liquid drops, and inserts (e.g., intraocular inserts). Such compositions can be formulated according to known methods.

[0261] Compositions intended for parenteral administration are typically provided as sterile aqueous or oily solutions or fine suspensions, or as sterile powders that can be finely pulverized for temporary reconstitution with sterile water for injection.

[0262] Examples of preparations for rectal or vaginal administration include pessaries and suppositories, which may be formed, for example, from moldable or waxy materials containing active compounds.

[0263] Compositions administered by inhalation may be in the form of inhalable powder compositions, liquids, or powder sprays, and may be administered in standard form using powder inhaler devices or aerosol dispensing devices. Such devices are well known. For inhalation administration, powder formulations typically contain an active compound along with an inert solid powder diluent (such as lactose).

[0264] The compounds of the present invention will generally be presented in unit dosage forms and will therefore generally contain a compound sufficient to provide the desired level of biological activity. For example, according to embodiment 4.1, a composition intended for oral administration may contain 2 mg to 200 mg of the active ingredient, more typically 10 mg to 100 mg, such as 12.5 mg, 25 mg or 50 mg.

[0265] Combination therapy It is envisioned that the compounds of embodiments 1.1 to 1.122 will be available as therapeutic agents alone or in combination with other therapeutic agents.

[0266] The compounds of this invention, designed to enhance mitochondrial bioenergetics by increasing OXPHOS capacity, are intended to be used in combination with the following: ● Replenish the NAD+ pool with potions; ● Inhibitors of pathways that deplete cellular and mitochondrial NAD+ pools; ● Regulators of mitochondrial biogenesis; ● Activator of retinoid X receptor-α (RXRα); ● AMPK agonists; ● SIRT1-activated compound (STAC); ● Mitochondrial-targeting protective compounds that reduce the production of toxic reactive oxygen species; ● Antioxidants; ● Vitamins and supplements; and ● Behavioral intervention.

[0267] Dosimetry The compounds of the present invention as defined in any one of embodiments 1.1 to 1.122 may be administered for a prolonged period to maintain a beneficial therapeutic effect, or may be administered for a short period of time. Alternatively, they may be administered in a pulsed or continuous manner.

[0268] The compounds of this invention will be applied in an effective amount, that is, an amount that effectively produces the desired therapeutic effect. For example, an "effective amount" may be the amount of a compound that improves disease symptoms and / or increases lifespan when applied to an individual.

[0269] The amount of the compound of the invention administered to an individual will depend on the type and severity of the disease or symptom, as well as the individual's characteristics (such as general health status, age, sex, weight, and tolerance to the drug). A person skilled in the art will be able to determine the appropriate dosage based on these and other factors.

[0270] The compound is typically administered to an individual who requires such administration, such as a human or animal individual (patient), preferably a human.

[0271] Ultimately, the amount of compound applied and the type of composition used will be appropriate to the nature of the disease or physiological condition being treated, and will be determined by the physician.

[0272] Diagnostic methods Before administering any of the compounds in embodiments 1.1 to 1.122, patients may be screened to determine whether the patient has or may have a disease or condition that is sensitive to treatment with compounds that can increase mitochondrial activity. Such patients may then be treated according to the methods described above.

[0273] For example, biological samples taken from a patient can be analyzed to determine whether the patient's existing or potential condition or disease is a condition or disease characterized by a genetic abnormality that leads to mitochondrial dysfunction. The term mitochondrial dysfunction encompasses both reduced mitochondrial production and decreased mitochondrial activity.

[0274] Mitochondrial dysfunction can be caused by exposure to certain environmental factors, such as occupational chemical mutagens, air pollution, and cigarette smoke, or by genetic abnormalities in mitochondrial (mtDNA) and nuclear DNA (nDNA). Mitochondrial dysfunction can affect any organ system and can occur at any age.

[0275] Diagnosing mitochondrial myopathy from patient tissue, skin, or serum samples involves techniques such as histological and immunohistochemical analysis: for example, staining frozen sections of skeletal muscle with a modified Gomori trichrome staining method to indicate the presence of broken red fibers (RRF); succinate dehydrogenase (SDH, complex II) histochemistry, used to detect mitochondrial aggregates formed due to mitochondrial OXPHOS dysfunction; and sequential COX / SDH histochemistry, which can be used to detect cytochrome c oxidase (COX, complex IV) negative fibers. Biochemical enzyme analysis using spectrophotometry to assess OXPHOS complexes (such as NADH: ubiquinone oxidoreductase, complex I; succinate: cytochrome c oxidoreductase, complex II; ubiquinone cytochrome c oxidoreductase, complex III; cytochrome c oxidase, complex IV) and blue native acrylamide PAGE (BN-PAGE) can both be used to detect mitochondrial dysfunction in patient tissue, skin, or serum. Mutations leading to mitochondrial myopathy can be present in either mtDNA or nDNA, and these mutations can be detected using next-generation sequencing (NGS), whole-exome sequencing (WES), whole-genome sequencing (WGS), or a combination of multi-gene targeted detection of candidate genes via NGS. For some patients with mitochondrial myopathy, determining the mtDNA copy number in muscle tissue using real-time PCR is helpful, or long-range PCR can be used to detect large-scale or multiple mtDNA deletions. Furthermore, mtDNA heterogeneity and deletions can be determined using NGS, real-time PCR, pyrosequencing, and long-range PCR.

[0276] In particular, mutations in mtDNA have been identified and linked to mitochondrial dysfunction in the following diseases:

[0277] Other diagnostic tests for mitochondrial myopathy include: measuring lactate levels at rest or after exercise; serum fibroblast growth factor 21 (FGF-21); and serum growth differentiation factor 15 (GDF-15). Integrating the information from these tests can help diagnose most patients with mitochondrial myopathy [Ahmed 2018].

[0278] In vitro applications As described above, the compounds of embodiments 1.1 to 1.122 can be used to improve mitochondrial activity in isolated cells. Such in vitro methods are envisioned for use in a variety of therapies involving the removal of cells from an individual, modification of the cells, and subsequent reintroduction of the cells into the individual. Examples of such therapies include CAR-T and CAR-NK therapies.

[0279] Therefore, an in vitro method for improving cellular mitochondrial activity is also provided, the method comprising contacting an effective amount of the compound according to any one of embodiments 1.1 to 1.122 with the cell.

[0280] Example Examples 1 to 56 The compounds in Examples 1 to 56 in Table 3 below are illustrative of the present invention.

[0281] The following examples are provided for illustrative purposes only and are not intended to limit the scope of the invention as described herein. For convenience, the following common abbreviations are used herein: - ACN stands for acetonitrile. - Boc is tert-butoxycarbonyl - CDI is 1,1′-carbonyldiimidazole - DCC is dicyclohexylcarbodiimide - DCM is dichloromethane - DMAP is 4-(dimethylamino)pyridine - DMF is N,N -Dimethylformamide - DMSO stands for dimethyl sulfoxide.

[0282] - EDC is 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide - h represents hours - HPLC stands for High Performance Liquid Chromatography.

[0283] - LCMS stands for Liquid Chromatography-Mass Spectrometry - min represents minutes - MTBE is methyl tert-butyl ether - NMR is nuclear magnetic resonance. - RT is the retention time - SFC stands for Supercritical Fluid Chromatography - TFA is trifluoroacetic acid - THF is tetrahydrofuran - TBDMS is tert-butyldi(methyl)silyl General method: Analytical methods Liquid chromatography-mass spectrometry LCMS - Method 1 LC-MS was performed on an Agilent 1260 Infinity II LC / MSD system equipped with either a DAD / ELSDAlltech 3300 or an Agilent DAD / ELSD G7102A 1290 Infinity II and an Agilent LC / MSDG6120B mass spectrometer. An Agilent Poroshell 120 SB-C18 2.7 µm 4.6 x 30 mm column was used, maintained at 60 °C. The column was initially held for 0.01 min in 1% acetonitrile (LC-MS grade) / high-purity water (Milli-Q) with 0.1% (v / v) formic acid, followed by a linear gradient of 1–100% acetonitrile over 1.5 min, and then held for 2.2 min in 100% acetonitrile at a flow rate of 3 mL / min.

[0284] LCMS - Method 2 LC-MS was performed on an Agilent 1260 Infinity II LC / MSD system equipped with either a DAD / ELSDAlltech 3300 or an Agilent DAD / ELSD G7102A 1290 Infinity II and an Agilent LC / MSDG6120B mass spectrometer. An Agilent Poroshell 120 SB-C18 2.7 µm 4.6 x 30 mm column was used, maintained at 60 °C. The column was initially held for 0.01 min in 1% acetonitrile (LC-MS grade) / high-purity water (Milli-Q) with 0.1% (v / v) formic acid, followed by a linear gradient of 1–100% acetonitrile over 5 min, and then held for 5.99 min in 100% acetonitrile at a flow rate of 1.5 mL / min.

[0285] LCMS - Method 3 LC-MS was performed on an Agilent 1290 series instrument with UV, ELSD 1290, and Agilent 6120 detectors, using a Waters Atlantis T3 (4.6 x 100 mm; 3 µm) column. The column was initially held for 2 min in 100% high-purity water (Milli-Q) with 0.05% (v / v) trifluoroacetic acid, followed by a linear gradient of 0–90% acetonitrile (LC-MS grade) over 3 min, and then held for 4 min in 90% acetonitrile at a flow rate of 1.0 mL / min.

[0286] LCMS - Method 4 LC-MS was performed on a UPLC-ESI-MS system equipped with a UV-VIS (Nexera UHPLC system LC40) and an MS detector (LCMS8045 triple quadrupole Shimadzu). A reversed-phase column (Zorbax XBD C18®, C18, 3.5 µm, 50 x 4.6 mm) was used and maintained at 40 °C. The column was initially held for 1 min in 5% acetonitrile (LC-MS grade) / high-purity water (Milli-Q) with 0.1% (v / v) formic acid, followed by a linear gradient of 5-95% acetonitrile over 2.5 min, and then isocratic held for 1.5 min in 95% acetonitrile at a flow rate of 0.7 mL / min.

[0287] NMR Unless otherwise stated, use a Bruker, Varian, or Agilent resonator. 1 1H nuclear magnetic resonance (NMR) spectroscopy was performed using a resonator operated at 400 MHz, 500 MHz, or 600 MHz, with the stated solvent, at approximately room temperature. In all cases, the NMR data were consistent with the proposed structure. Characteristic chemical shifts (δ) were given in parts per million, using conventional abbreviations to identify major peaks: e.g., s, singlet; d, doublet; t, triplet; q, quartet; dd, double doublet; dt, double triplet; m, multiplet; br, broad peak.

[0288] Purification methods Preparative reversed-phase HPLC conditions Preparative HPLC purification was performed using an Agilent Technologies 1260 Infinity LC / MSD system via high-performance liquid chromatography (HPLC). Standard HPLC methods were employed using a Chromatorex C18 (19 mm x 100 mm, 5 µm) column at 25 °C; the flow rate was 30 mL / min (4 mL / min injection pump). Appropriate focusing gradients were selected based on the acetonitrile and methanol solvent system under acidic or basic conditions. The modifiers used under acidic / basic conditions were trifluoroacetic acid, HCl, or NH3 32% solution (0.3% v / v), respectively.

[0289] Typical gradient is:

[0290] Samples were loaded onto an ACD and analyzed using a DAD / ELSD Alltech 3300 (210–400 nm) and an Agilent / LC / MSD G6120B mass spectrometer (in SIM and TIC modes), and collected using an Agilent 1200 series HPLC G1364B preparative fraction collector. Fractions containing the target ions were concentrated under reduced pressure, transferred to pre-weighed flasks, lyophilized, and analyzed using LCMS and NMR techniques.

[0291] HPLC conditions 1: Mobile phase 5-5-25%, 0-1-5 min H2O / ACN / 0.1% TFA; Column Phenyl SMB100-5 100x19 mm 5µm.

[0292] HPLC condition 2: Mobile phase 50-75%, 0-5 min H2O / ACN; Column Chromatorex 18 SMB100-5T 100x19 mm 5µm HPLC condition 3: Mobile phase 5-30%, 0-5 min H2O / ACN / 0.1% TFA; Column Phenyl SMB100-5 100x19 mm 5µm HPLC condition 4: Mobile phase 5-5-20%, 0-2-5 min H2O / ACN / 0.1% TFA; Column Chromatorex 18SMB100-5T 100x19 mm 5µm synthesis This document describes several methods for chemically synthesizing the azabicyclic dimer compounds of this application. These and / or other well-known methods can be modified and / or adapted in various ways to facilitate the synthesis of other compounds within the scope of this application and the claims. Such alternative methods and modifications should be understood as being within the spirit and scope of this application and the claims. Therefore, it should be understood that the methods set forth in the following descriptions, schemes, and examples are intended for illustrative purposes and should not be construed as limiting the scope of this disclosure.

[0293] Start 2-(tert-butoxycarbonyl)-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (I-VII).

[0294] Intermediate II. 1R, 4S, 5S )-2-(tert-butoxycarbonyl)-2-azabicyclo[2.1.1]hexane-5-carboxylic acid After 0.1 h, 2-(tert-butoxycarbonyl)-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (CAS1279894-35-7, 81.7 g, 0.35 mol) in THF (2000 mL) was added to the stirred solution. R -( + 1-Phenylacetylamine (47 mL, 0.37 mol, 1.05 equivalents). The mixture was then filtered, and the solid was collected and recrystallized from THF. The resulting material was further recrystallized from ACN to obtain a solid. The solid was suspended in ethyl acetate (200 mL), and 2... N Hydrochloric acid (200 mL). The organic phase was separated, dried (Na₂SO₄), and concentrated under reduced pressure to give the title compound (20 g, 49% yield) as a pale yellow oil. Chiral SFC (Method 1) RT 3.28 min (99%) ee ). 1 H NMR (400 MHz, CDCl3): δ4.60 (d, 1H), 3.60 (d, 1H), 3.30 (d, 1H), 3.32 - 3.29 (m, 1H), 2.82 - 2.80(m, 1H), 1.81 - 1.78 (m, 1H), 1.42 (d, 9H), 1.38 (m, 1H).

[0295] Intermediate III. 1S, 4R, 5R )-2-(tert-butoxycarbonyl)-2-azabicyclo[2.1.1]hexane-5-carboxylic acid Add to a stirred solution of 2-(tert-butoxycarbonyl)-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (CAS 1279894-35-7, 290 g, 1.28 mol) in THF (1500 mL) S-(-)-1-phenylethylamine (75 g, 0.62 mol, 0.5 equivalents) was stirred at room temperature for 18 h, the mixture was filtered, and the solid was recrystallized from THF and subsequently from ACN. The resulting solid was added to ethyl acetate (600 mL), and 2... N HCl aqueous solution (600 mL). Separate the organic phase, dry (Na2SO4), and concentrate under reduced pressure to obtain a yellow oil. 1S, 4R, 5R 2-(tert-butoxycarbonyl)-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (42 g, 28% yield). Chiral SFC (Method 1) RT 4.06 min (99%) ee ). 1 ¹H NMR (400 MHz, CDCl₃): δ 4.60 (d, 1H), 3.60 (d, 1H), 3.30 (d, 1H), 3.30 - 3.26 (m, 1H), 2.85 - 2.74 (m, 1H), 1.83 - 1.79 (m, 1H), 1.42 (d, 9H), 1.40 - 1.37 (m, 1H). No exchangeable protons were observed.

[0296] Intermediate IV. 2-(tert-butoxycarbonyl)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylic acid Step A Synthesis (3) E )-4-[(2-methylprop-2-en-1-yl)amino]but-3-en-2-one Add (3) to a solution of 2-methylprop-2-en-1-amine (20 g, 281 mmol, 1.05 equivalents) in THF (500 mL). E )-4-methoxybut-3-en-2-one (27 g, 268 mmol). After stirring at room temperature for 18 h, the reaction mixture was concentrated under reduced pressure to give a pale yellow oil (3-) E 4-[(2-methylprop-2-en-1-yl)amino]but-3-en-2-one (40 g, 97% yield) was used in the next step without further purification. LCMS (Method 1) RT = 0.85 min, m / z :[ESI + 140.2 (M+H) + . 1¹H NMR (400 MHz, CDCl₃): δ 9.78 (br s, 1H), 9.78 (m, 1H), 6.58 - 6.53 (m, 1H), 4.98 - 4.96 (m, 1H), 4.83 (s, 2H), 3.63 (d, 2H), 2.01 (s, 3H), 1.68 (s, 3H). No exchangeable protons were observed.

[0297] Step B synthesis N -(2-Methylpropyl-2-en-1-yl)- N -[(1 E 3-O-But-1-en-1-yl]tert-butyl carbamate Towards( 3E 4-[(2-methylprop-2-en-1-yl)amino]but-3-en-2-one (40 g, 287 mmol) was dissolved in THF (1000 mL) at 0 °C with sodium tert-butoxide (27 g, 284 mmol, 1.1 equivalents). Then, di-tert-butyl dicarbonate (56 g, 258 mmol, 1.0 equivalents) was added, and the mixture was stirred for another 3 h. The solvent was then removed under reduced pressure, MTBE (400 mL) was added, and the organic layer was washed with brine. The combined organic layers were dried (Na₂SO₄) and concentrated under reduced pressure. The residue was purified by rapid column chromatography to give a pale yellow oil. N -(2-Methylpropyl-2-en-1-yl)- N -[( 1E 3-O-But-1-en-1-yl]tert-butyl carbamate (52 g, 84% yield). LCMS (Method 1) RT = 1.29 min, m / z :[ESI + 140.2 (M+H-56) + . 1 H NMR (400 MHz, CDCl3): δ 8.18 - 8.16 (m, 1H), 5.48 -5.45 (m, 1H), 4.88 (s, 1H), 4.69 (s, 1H), 4.09 (s, 2H), 2.23 (s, 3H), 1.71(s, 3H), 1.53 (s, 9H).

[0298] Step C Synthesis of tert-butyl-5-acetyl-4-methyl-2-azabicyclo[2.1.1]hexane-2-carboxylate Will N -(2-Methylpropyl-2-en-1-yl)- N -[( 1E10.0 g (42 mmol) of tert-butyl 3-oxobut-1-en-1-yl]carbamate was dissolved in ACN (1000 mL) and irradiated with a UV lamp (350 nm) in a flow reactor for 3.3 h at a flow rate of 2.5 mL / min. The reaction mixture was then concentrated under reduced pressure and purified by rapid column chromatography to give 4.0 g (47% yield) of tert-butyl-5-acetyl-4-methyl-2-azabicyclo[2.1.1]hexane-2-carbamate as a pale yellow oil. 1 H NMR (400 MHz, CDCl3): δ 4.57 - 4.55 (m, 1H), 3.27 - 3.25 (m, 1H), 3.02- 2.97 (m, 1H), 2.40 (s, 1H), 2.05 (s, 3H), 1.49 - 1.45 (m, 2H), 1.43 (s,9H), 1.37 (s, 3H).

[0299] Step D Synthesis of 2-[(tert-butoxy)carbonyl]-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylic acid Bromine (1.67 g, 10 mmol, 5.0 equivalent) was added to a solution of sodium hydroxide (836 mg, 21 mmol, 10.0 equivalent) in water (4 mL) at 0 °C with stirring. The reaction mixture was stirred for 0.5 h, followed by the addition of a solution of tert-butyl-5-acetyl-4-methyl-2-azabicyclo[2.1.1]hexane-2-carboxylic acid (500 mg, 2.1 mmol) in dioxane (2 mL). The mixture was stirred at room temperature for 4 h. The reaction mixture was then quenched with sodium sulfite solution, and the resulting mixture was washed twice with diethyl ether. The organic layer was washed with brine, dried (Na₂SO₄), and concentrated under reduced pressure to give 2-[(tert-butoxy)carbonyl]-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (400 mg, 93% yield) as a colorless solid. LCMS (Method 1) RT = 1.15 min, m / z : [ESI - 240.2 (MH) - . 1¹H NMR (400 MHz, CDCl₃): δ 4.50 (br s, 1H), 3.39 - 3.36 (m, 1H), 3.07 - 3.05 (m, 1H), 2.48 (s, 1H), 1.58 - 1.55 (m, 1H), 1.43 (s, 9H), 1.44 - 1.41 (m, 1H), 1.35 (s, 3H). No exchangeable protons were observed.

[0300] Intermediate V. ( 1R, 4S, 5S )-2-(tert-butoxycarbonyl)-4-methyl-2-azabicyclo[2.1.1]hexane-5- Formic acid Add ( ) to a reflux-stirred suspension of 2-(tert-butoxycarbonyl)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (302 g, 0.68 mol) in ACN (2 L) S A solution of phenylglycine (94 g, 0.69 mol, 1.01 equivalents) in ACN (2 L) was prepared. The reaction mixture was stirred under reflux for 0.5 h and then cooled to room temperature. The resulting suspension was filtered, the filter cake was washed with ACN and dried under reduced pressure to give phenylglycine salt (1... R 4 S 5 S 2-(tert-butoxycarbonyl)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylic acid t (110 g, 85% yield). Chiral HPLC: 99.4% ee .

[0301] To (1) R 4 S 5 S A stirred suspension of 2-(tert-butoxycarbonyl)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylic acid phenylglycanol salt (51 g, 0.13 mol) in DCM (1 L) was supplemented with an aqueous solution of potassium hydrogen sulfate (21 g, 0.15 mol, 500 mL, 1.1 eq). The two-phase mixture was stirred for 0.2 h, and the phases were then separated. The aqueous layer was extracted with DCM, and the combined organic layers were dried (Na2SO4), filtered, and concentrated under reduced pressure to give a grayish-white solid ( 1R, 4S, 5S 2-(tert-butoxycarbonyl)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (30 g, 90% yield). LCMS (Method 3) RT = 1.65 min, m / z : [ESI + 242.2 (M+H) + .

[0302] Intermediate VI. ( 1S, 4R, 5R )-2-(tert-butoxycarbonyl)-4-methyl-2-azabicyclo[2.1.1]hexane-5- Formic acid After 1 hour, towards splitting ( 1R, 4S, 5S The mother liquor of 2-(tert-butoxycarbonyl)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (14.7 g, 0.03 mol) was added to a refluxed, stirred solution of ACN (100 mL). R A solution of 3.85 g, 0.03 mol, 1 equivalent of phenylglycine was prepared in 100 mL of ACN. After the addition was complete, the reaction mixture was stirred under reflux for 0.5 h, cooled to room temperature, and stirred overnight. The reaction mixture was cooled in an ice bath for 1 h, and the resulting suspension was filtered. The filter cake was washed with ACN and dried under reduced pressure to give phenylglycine salt ( 1S, 4R, 5R 2-(tert-butoxycarbonyl)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (6.32 g, quantitative). Chiral HPLC: 99.9% ee .

[0303] Towards( 1S, 4R, 5R A stirred suspension of 2-(tert-butoxycarbonyl)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylic acid phenylglycanol salt (6.32 g, 0.02 mol) in DCM (120 mL) was supplemented with an aqueous solution of potassium hydrogen sulfate (2.5 g, 0.02 mol, 73 mL, 1.1 equivalents). The two-phase mixture was stirred for 0.2 h, and the phases were then separated. The aqueous layer was extracted with DCM, and the combined organic layers were dried (Na2SO4), filtered, and concentrated under reduced pressure to give a grayish-white solid ( 1S, 4R, 5R 2-(tert-butoxycarbonyl)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (3.59 g, 90% yield). Chiral HPLC: 100% ee LCMS (Method 3) RT = 1.64 min, m / z : [ESI + 242.2 (M+H) + . 1¹H NMR (400MHz, CDCl₃): δ 4.52 (br s, 1H), 3.40 - 3.37 (m, 1H), 3.07 - 3.05 (m, 1H), 2.50 (s, 1H), 1.58 - 1.56 (m, 1H), 1.45 (s, 9H), 1.45 - 1.41 (m, 1H), 1.37 (s, 3H). No exchangeable protons were observed.

[0304] Intermediate VII: 2-(tert-butoxycarbonyl)-4-phenyl-2-azabicyclo[2.1.1]hexane-5-carboxylic acid Step A synthesis( E )-4-((2-phenylallyl)amino)but-3-en-2-one Add ( ) to a 0°C stirred solution of 2-phenylprop-2-en-1-amine (1.23 g, 9.2 mmol) in diethyl ether (30 mL) E )-4-methoxybut-3-en-2-one (0.93 mL, 9.2 mmol, 1.0 equivalent). The resulting mixture was stirred at room temperature for 16 h. Afterward, the mixture was cooled to 0 °C and additional ( E )-4-methoxybut-3-en-2-one (0.46 mL, 4.6 mmol, 0.5 equivalents). The resulting mixture was stirred at room temperature for 20 h. The mixture was concentrated under reduced pressure to give a yellow oil. E )-4-((2-phenylallyl)amino)but-3-en-2-one, which is used directly in the next step.

[0305] Step B synthesis( E 3-O-But-1-en-1-yl)(2-Phenylacetyl)carbamate tert-butyl Towards( E 1.87 g (9.2 mmol) of 4-((2-phenylallyl)amino)but-3-en-2-one (30 mL) was added to a stirred solution at 0 °C in DCM, along with 2.33 mL (10.2 mmol, 1.1 equivalents) of di-tert-butyl dicarbonate and DMAP (56 mg, 0.05 mmol, 0.05 equivalents). The resulting mixture was stirred at room temperature for 16 h. Subsequently, the reaction mixture was concentrated under reduced pressure and purified directly by rapid column chromatography to give a yellow oil (…). E )-(3-oxobut-1-en-1-yl)(2-phenylallyl) tert-butyl carbamate (1.15 g, 42% yield, via 2 steps). 1 H NMR (400 MHz, CDCl3) δ 8.20 (d, J= 14.5 Hz, 1H), 7.39 - 7.27 (m, 5H), 5.53 (d, J = 14.5 Hz, 1H), 5.36 (t, J = 1.7Hz, 1H), 4.94 (t, J = 1.9 Hz, 1H), 4.55 (t, J = 1.8 Hz, 2H), 2.21 (s, 3H), 1.52 (s, 9H).

[0306] Step C Synthesis of tert-butyl 5-acetyl-4-phenyl-2-azabicyclo[2.1.1]hexane-2-carboxylate Will( E 3-(3-oxobut-1-en-1-yl)(2-phenylallyl) tert-butyl carbamate (380 mg, 1.26 mmol, 1.00 equivalent) was dissolved in acetonitrile (12 mL). [Ir(dF(CF3)ppy)2(dtbbpy)][PF6] (3.2 mg, 0.033 mmol, 0.01 equivalent) was weighed into a photochemical reaction vial. 3.2 mL of the prepared ACN solution was added to the vial, and N2 was bubbled through the mixture for 25 min. This process was repeated four times to obtain four batches. The four vials were sealed and placed in a fan-cooled photochemical reactor, irradiated with a blue LED at 456 nm for 5 h. After the starting material was completely consumed, the vials were combined, and the mixture was concentrated under reduced pressure. The residue was purified by rapid column chromatography to obtain tert-butyl 5-acetyl-4-phenyl-2-azabicyclo[2.1.1]hexane-2-carboxylate (115 mg, 30% yield), which was a yellow oil. 1 H NMR (400 MHz, CDCl3) δ 7.39 - 7.30 (m, 3H), 7.25 - 2.22 (m, 2H), 4.59 (d, J = 2.1 Hz, 1H), 3.84 (dd, J = 8.9, 1.7 Hz, 1H), 3.50 (d, J = 8.9 Hz, 1H), 3.22 (d, J = 7.4 Hz, 1H), 2.99 (dt, J = 7.7, 1.9 Hz, 1H), 1.88 (t, J = 7.6 Hz, 1H), 1.78 (s, 3H), 1.49 (s, 9H).

[0307] Step D Synthesis of 2-(tert-butoxycarbonyl)-4-phenyl-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (V) Bromine (43 µL, 0.84 mmol, 4.0 equivalents) was added to a solution of sodium hydroxide (84 mg, 2.10 mmol, 10.0 equivalents) in water (1.3 mL) at 0 °C with stirring. This was followed by the addition of a solution of 5-acetyl-4-phenyl-2-azabicyclo[2.1.1]hexane-2-carboxylic acid tert-butyl ester (63 mg, 0.20 mmol) in 1,4-dioxane (0.4 mL). The resulting mixture was stirred and heated to room temperature over 16 hours. The reaction mixture was diluted with water, washed with diethyl ether, acidified to pH 2–3 with aqueous hydrochloric acid, and extracted with ethyl acetate. The combined organic layers were dried (Na₂SO₄), filtered, and concentrated under reduced pressure to give 2-(tert-butoxycarbonyl)-4-phenyl-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (a mixture of diastereomers, 45 mg, 71% yield) as a grayish-white solid. 1 H NMR (400 MHz, DMSO- d 6) δ 12.31 (br s, 1H), 7.40 - 7.24(m, 5H), 4.37 - 4.36 (m, 1H), 3.65 - 3.63 (m, 1H), 3.25 - 3.21 (m, 1H), 3.04- 3.00 (m, 2H), 1.92 - 1.88 (m, 1H), 1.43 (s, 9H).

[0308] 1,1'-Sulfoylbis(2-methylprop-2-ol) Step A Synthesis of methyl 2-[(2-methoxy-2-oxoethyl)thio]acetate At -5 °C, thionyl chloride (2.38 g, 19.9 mmol, 3.0 equivalent) was added dropwise to a stirred solution of 2-[(carboxymethyl)thio]acetic acid (1.0 g, 6.66 mmol) in methanol. The mixture was stirred overnight at room temperature. The resulting mixture was concentrated under reduced pressure, and the residue was diluted with MTBE (200 mL). The organic layer was washed with sodium carbonate and brine, dried (Na₂SO₄), and concentrated under reduced pressure to give methyl 2-[(2-methoxy-2-oxoethyl)thio]acetate (2.6 g, 73% yield) as a colorless oil. 1 H NMR (500 MHz, CDCl3) δ 3.75 (s, 6H), 3.41 (s, 4H).

[0309] Step B Synthesis of 1-[(2-hydroxy-2-methylpropyl)thio]-2-methylprop-2-ol At -5°C, for 30 minutes, a solution of methyl 2-[(2-methoxy-2-oxoethyl)thio]acetate (2.6 g, 14.6 mmol) in THF (20 mL) was added to a stirred solution of magnesium chloride (8.73 g, 116.7 mmol, 8.0 equivalent) in THF (250 mL). The resulting mixture was stirred overnight at room temperature. Subsequently, a saturated aqueous solution of ammonium chloride was added to the mixture, and the organic phase was separated. The aqueous layer was extracted with freshly prepared THF, and the combined organic layers were concentrated under reduced pressure. The residue was dissolved in hexane (100 mL), dried (Na2SO4), concentrated under reduced pressure, and purified by preparative HPLC to give 1-[(2-hydroxy-2-methylpropyl)thio]-2-methylprop-2-ol (396 mg, 15.2% yield) as a yellow oil. 1 ¹H NMR (500MHz, CDCl₃) δ 2.28 (s, 4H), 1.26 (s, 12H), no exchangeable protons observed.

[0310] Step C Synthesis of 1-(2-hydroxy-2-methylpropanesulfonyl)-2-methylprop-2-ol At 0 °C, 3-chlorobenzene-1-peroxycarboxylic acid (1.74 g, 10.1 mmol, 2 equivalents) was added to a stirred solution of 1-[(2-hydroxy-2-methylpropyl)thio]-2-methylpropane-2-ol 3 (396 mg, 5.05 mmol) in DCM (15 mL). The resulting mixture was stirred overnight at room temperature. Subsequently, the mixture was concentrated under reduced pressure, and the residue was purified by preparative HPLC to give 1-(2-hydroxy-2-methylpropanesulfonyl)-2-methylpropane-2-ol (225 mg, 21.2% yield) as a colorless oil. 1 HNMR (500 MHz, CDCl3) δ 2.47 (s, 4H), 1.48 (s, 12H), no exchangeable protons observed.

[0311] Option 1 In one method (Scheme 1), the compound is prepared by: in a polar aprotic solvent (such as THF or dioxane), in the presence of a nucleophilic base (such as DMAP), and with the aid of a coupling agent (such as DCC or DIC). N,N'In the Steglich esterification of diisopropylcarbodiimide, a carboxylic acid derivative of general formula (20) reacts with a dihydroxy compound of general formula (21). The reaction is generally carried out at room temperature. After reaction treatment typically by liquid-liquid extraction, the reaction product is purified by rapid column chromatography, reversed-phase preparative HPLC, or recrystallization. Subsequently, it is purified by general... N -Boc deprotection steps, such as reaction with TFA or HCl in a polar solvent (e.g., DCM or diethyl ether), are used to prepare compounds of general formula (22). After reaction treatment, which is usually carried out by ion exchange purification or liquid-liquid extraction, the reaction products are purified by rapid column chromatography, reversed-phase preparative HPLC, or recrystallization.

[0312] Example 1: Propane-1,3-dimethylbis(2-azabicyclo[2.1.1]hexane-5-carboxylate) trifluoroacetate Step A Synthesis of 2,2'-di-tert-butyl5,5'-(propane-1,3-diyl)bis(2-azabicyclo[2.1.1]hexane-2,5-di Formate) To 2-[(tert-butoxy)carbonyl]-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (1.50 g, 6.6 mmol, 2.2 equivalents), N , N '-Dicyclohexylmethyldiimide (2.04 g, 9.90 mmol, 3.3 equivalents) and N , N 1,2'-Dimethylpyridin-4-amine (161 mg, 1.32 mmol, 0.44 equivalents) was added to a stirred solution of THF (15 mL) with propane-1,3-diol (226 mg, 2.97 mmol, 1.0 equivalents), and the mixture was stirred at room temperature for 18 h. The solvent was then concentrated under reduced pressure, and hexane (50 mL) was added to the residue. The resulting suspension was filtered, and the filtrate was washed with aqueous citric acid, aqueous sodium bicarbonate solution, and brine. The combined organic layers were dried (Na₂SO₄), filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (condition 2) to give 2,2'-di-tert-butyl as a colorless oil. 5,5’- (propane-1,3-diyl)bis(2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate) (0.32 g 22% yield).

[0313] Step B Synthesis of propane-1,3-dimethylbis(2-azabicyclo[2.1.1]hexane-5-carboxylate) trifluoroacetate 2,2'-di-tert-butyl 5,5’-(Propane-1,3-diyl)bis(2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate) (0.32 g, 0.66 mmol) was dissolved in TFA (3 mL), and the solution was stirred at room temperature for 0.25 h. The reaction mixture was then concentrated under reduced pressure. The resulting residue was dissolved in water (10 mL), washed with DCM, and concentrated under reduced pressure. The residue was purified by preparative HPLC (condition 1) to give propane-1,3-diylbis(2-azabicyclo[2.1.1]hexane-5-carboxylate) trifluoroacetate (0.17 g, 50% yield) as a grayish-white solid. LCMS (Method 2) RT = 0.37 min, m / z : [ESI - 295.16 (M+H) + . 1 H NMR (400 MHz, D2O) δ 4.32 (dt, J = 5.9, 1.7 Hz, 2H),4.16 - 4.02 (m, 4H), 3.43 - 3.30 (m, 4H), 3.21 - 3.10 (m, 4H), 2.05 - 1.98(m, 2H), 1.95 - 1.88 (m, 2H), 1.32 (dd, J = 9.4, 1.1 Hz, 2H), no exchangeable protons were observed.

[0314] Scheme 2. A general scheme for the synthesis of bis(2-azabicyclo[2.1.1]hexane-5-carboxylate).

[0315] Step A At room temperature, 2-(tert-butoxycarbonyl)-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (I-VII) (2.4 mmol, 2.2 equivalents), DCC (595 mg, 2.88 mmol, 2.7 equivalents), and DMAP (587 mg, 4.81 mmol, 4.5 equivalents) were dissolved in THF (10 mL). After stirring for 5 min, the corresponding diol (1.1 mmol, 1.0 equivalent) was added, and the reaction mixture was stirred at room temperature for 18 h. The solvent was concentrated under reduced pressure, and diethyl ether (50 mL) was added to the residue. The resulting suspension was filtered. The filtrate was washed with aqueous citric acid, aqueous NaHCO3, and brine, dried (Na2SO4), filtered, and concentrated under reduced pressure. All intermediates were separated by preparative HPLC.

[0316] Step B Each intermediate (1.0 equivalent) was dissolved in DCM (1 mL / mmol) and cooled to 0 °C, followed by dropwise addition of TFA (5.0 equivalent). The solution was stirred overnight at room temperature. The resulting mixture was concentrated under reduced pressure, and the residue was purified by preparative HPLC to give the final product as a TFA salt.

[0317] The following compounds were prepared according to Scheme 1 of the above methods:

[0318] Example 35. (1-Methylazacyclobutane-3,3-diyl)bis(methylene)bis(2-azabicyclo[2.1.1]hexane- 5-Carbamate) Trifluoroacetate Step A Synthesis of 3,3-bis(hydroxymethyl)-azacyclobutane-1-carboxylic acid methyl ester Potassium carbonate (1.11 g, 8.01 mmol, 2.5 equivalents) was added to a stirred solution of [3-(hydroxymethyl)azacyclobutane-3-yl]methanol hydrochloride (492 mg, 3.2 mmol) in water / THF (5 mL / 10 mL) at 0 °C. The reaction mixture was stirred for 10 min, followed by the addition of benzoyl chloroformate (574 mg, 3.36 mmol, 1.05 equivalents) in THF (6 mL). The resulting mixture was then stirred at room temperature for 4 h. The mixture was then poured into water and extracted with ethyl acetate. The combined organic layers were washed with brine, dried (Na₂SO₄), filtered, concentrated under reduced pressure, and purified by preparative HPLC to give benzoyl 3,3-bis(hydroxymethyl)azacyclobutane-1-carboxylate (600 mg, 99% yield) as a colorless oil. LCMS (Method 1) RT = 0.86 min, m / z : [ESI + 252.12 (M+H) + .

[0319] Step B Synthesis of 5,5'-[{1-[(benzoxy)carbonyl]azacyclobutane-3,3-diyl}bis(methylene)]2,2'-di tert-Butyl di(2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate) To 2-[(tert-butoxy)carbonyl]-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (1.50 g, 6.6 mmol), N , N '-Dicyclohexylmethyldiimide (3.54 g, 17.2 mmol, 2.6 equivalents) and N , N Dimethylpyridine-4-amine (8.0 mg, 0.07 mmol, 0.01 equivalent) was added to a stirred solution of 3,3-bis(hydroxymethyl)azacyclobutane-1-carboxylic acid (829 mg, 3.3 mmol, 0.5 equivalent) in anhydrous THF (200 mL) at 0 °C. The reaction mixture was stirred overnight at room temperature. The resulting precipitate was filtered, and the organic phase was concentrated under reduced pressure. The residue was purified by rapid column chromatography to give 5,5'-[{1-[(benzoxy)carbonyl]azacyclobutane-3,3-diyl}bis(methylene)]2,2'-di-tert-butyldi(2-azabicyclo[2.1.1]hexane-2,5-dicarboxylic acid ester) (1.1 g, 28% yield) as a grayish-white solid. LCMS (Method 1) RT = 1.59 min, m / z : [ESI + 570.30 (M-100+H) + .

[0320] Step CD Synthesis of 2,2'-di-tert-butyl5,5'-[(1-methylazacyclobutane-3,3-diyl)bis(methylene)]bis(2-aza Heterobicyclo[2.1.1]hexane-2,5-dicarboxylate) To a stirred solution of 5,5'-[{1-[(benzoxy)carbonyl]azacyclobutane-3,3-diyl}bis(methylene)]2,2'-di-tert-butyldi(2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate) (500 mg, 0.75 mmol) in methanol (30 mL), 10% Pd / C (8 mg) and formaldehyde (22.4 mg, 0.75 mmol, 1 equivalent) were added. The resulting mixture was hydrogenated at 50 °C (50 atm) and stirred in an autoclave for 24 h. The catalyst was filtered, and the solvent was concentrated under reduced pressure to obtain crude 2,2'-di-tert-butyl-5,5'-[(1-methylazacyclobutane-3,3-diyl)bis(methylene)]bis(2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate) (490 mg) as a yellow oil, which was used in the next step without further purification. LCMS (Method 2) RT = 0.86 min, m / z : [ESI + 550.31 (M+H) + .

[0321] Step E Synthesis of (1-methylazacyclobutane-3,3-diyl)bis(methylene)bis(2-azabicyclo[2.1.1]hexane-5- Formate (trifluoroacetate) See step B of scheme 2. The crude material was purified by preparative HPLC (method 4) to obtain a yellow oil of (1-methylazacyclobutane-3,3-diyl)bis(methylene)bis(2-azabicyclo[2.1.1]hexane-5-carboxylate) trifluoroacetate (298 mg, 47% yield, in two steps). LCMS (method 2) RT = 0.11 min, m / z : [ESI + 350.20 (M+H) + . 1 H NMR (400 MHz, DMSO- d 6) δ 10.50 (s, 1H), 9.50 (br s, 2H), 9.00 (br s,2H), 4.46 (d, J = 6.1 Hz, 2H), 4.30 - 4.05 (m, 6H), 3.92 (d, J = 14.4 Hz, 2H), 3.43 - 3.22 (m, 6H), 3.18 - 3.07 (m, 2H), 2.85 (d, J = 4.1 Hz, 3H), 1.97 (q, J =8.7, 8.2 Hz, 2H), 1.31 (d, J = 8.6 Hz, 2H).

[0322] Example 36. (1,1-oxo-1λ) 6 -thiamethoxane-4,4-diyl)bis(methylene)bis(2-azabicyclo[2.1.1]hexane (Alkyl-5-carboxylate) trifluoroacetate Step A Synthesis of (thiazide-4,4-diyl)diethanol Lithium aluminum hydride (689 mg, 20.3 mmol, 5 equivalents) was added to a stirred solution of 4-[(tert-butoxy)carbonyl]thiane-4-carboxylic acid (1.00 g, 4.06 mmol) in THF (20 mL) at 0 °C. The mixture was stirred at room temperature for 16 h. The reaction mixture was then cooled to 0 °C, and a solution of NaOH (275 mg) in water (2.7 mL) was added. The resulting precipitate was filtered, and the solvent was concentrated under reduced pressure. The residue was dissolved in DCM (30 mL), washed with water, dried (Na₂SO₄), and filtered. The solvent was concentrated under reduced pressure to give a yellow oil. thiamethoxane -4,4-dimethyl)diethanol (500 mg, 76% yield). 1 H NMR (400 MHz, CDCl3) δ 3.65 (d, J = 5.5 Hz, 2H), 3.61 (s, 4H), 2.61 - 2.49 (m, 4H), 1.74 - 1.68 (m, 4H).

[0323] Step B Synthesis of 2-di-tert-butyl5,5'-[thiapan-4,4-diylbis(methylene)]bis(2-azabicyclo[2.1.1]hexane- 2,5-Dicarboxylate Add (thiazide-4,4-diyl)diethanol (195 mg, 1.2 mmol) and 2-[(tert-butoxy)carbonyl]-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (600 mg, 2.64 mmol, 2.2 equivalents) to a stirred solution of (thiazide-4,4-diyl)diethanol (195 mg, 1.2 mmol) and 2-[(tert-butoxy)carbonyl]-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (600 mg, 2.64 mmol, 2.2 equivalents) in anhydrous DCM (25 mL) at 0 °C. N , N '-Dicyclohexyl-methyldiimide (594 mg, 2.88 mmol, 2.4 equivalents) and N , N -Dimethylpyridine-4-amine (88 mg, 0.72 mmol, 0.6 equivalents). The reaction mixture was stirred overnight at room temperature. The resulting precipitate was then filtered. The organic phase was washed with water, dried (Na₂SO₄), and filtered. The solvent was concentrated under reduced pressure, and the residue was purified by preparative HPLC to give 2-di-tert-butyl-5,5'-[thiapan-4,4-diylbis(methylene)]bis(2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate) (188 mg, 31% yield) as a yellow oil. LCMS (Method 2) RT = 1.53 min, m / z : [ESI + 581.33 (M+H) + .

[0324] Step C Synthesis of 2,2'-di-tert-butyl5,5'-[(1,1-oxo-1λ)] 6 -thiazide-4,4-diyl)bis(methylene)]bis(2-nitrogen Heterobicyclo[2.1.1]hexane-2,5-dicarboxylate) 3-Chlorobenzene-1-peroxycarboxylic acid (123 mg, 0.71 mmol, 2.2 equivalences) was added to a stirred solution of 2-di-tert-butyl5,5'-[thiapan-4,4-diylbis(methylene)]bis(2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate) (188 mg, 0.32 mmol) in DCM (10 mL) at 0 °C. The reaction mixture was stirred overnight at room temperature. The solution was then washed with a saturated aqueous solution of sodium bicarbonate and brine. The combined organic layers were dried (Na₂SO₄), filtered, and concentrated under reduced pressure to give a beige oil of 2,2'-di-tert-butyl5,5'-[(1,1-oxo-1λ) 6 [-thiamethoxane-4,4-diyl)bis(methylene)]bis(2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate) (175 mg, 99% yield), was used in the next step without further purification. LCMS (Method 2) RT = 1.00 min, m / z : [ESI + 613.27 (M+H) + .

[0325] Step D Synthesis of (1,1-oxo-1λ) 6 -thiamethoxane-4,4-diyl)bis(methylene)bis(2-azabicyclo[2.1.1]hexane-5- Formate (trifluoroacetate) The title compound was prepared using a procedure similar to that outlined in step B of scheme 2. The crude material was purified by preparative HPLC (method 3) to obtain a beige oil of (1,1-oxo-1λ). 6 1,4-Thiamethane-4,4-diyl)bis(methylene)bis(2-azabicyclo[2.1.1]hexane-5-carboxylate) trifluoroacetate (88 mg, 48% yield). LCMS (Method 2) RT = 0.20 min, m / z : [ESI + 413.17 (M+H) + . 1 H NMR (400 MHz, CD3OD) δ 4.42 (dt, J = 6.1, 1.7 Hz,2H), 4.21 - 4.16 (m, 4H), 3.59 - 3.47 (m, 2H), 3.43 (d, J = 10.1 Hz, 2H), 3.31- 3.23 (m, 4H), 3.13 (t, J= 6.1 Hz, 4H), 2.23 - 1.98 (m, 6H), 1.44 (d, J = 9.2Hz, 2H), no exchangeable protons were observed.

[0326] Example 37. Ethane-1,2-dimethylbis[(1R,4S,5S)-2-azabicyclo[2.1.1]hexane-5-carboxylate] hydrochloric acid Salt Step A Synthesis of 2,2'-di-tert-butyl5,5'-ethane-1,2-dimethylbis[(1R,4S,5S)-2-azabicyclo[2.1.1]hexane [Alkyl-2,5-dicarboxylate] Add ( ) to a stirred solution of 1,1'-carbonyldiimidazole (8.0 g, 49.2 mmol, 2.2 equivalents) in DMF (50 mL) 1R, 4S, 5S A solution of 2-(tert-butoxycarbonyl)-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (11.0 g, 48.2 mmol, 2.2 equivalents) in DMF (55 mL) was added, and the reaction mixture was stirred at room temperature for 1 hour. Then, ethane-1,2-diol (1.4 g, 22.4 mmol, 1 equivalent) and 2,3,4,6,7,8,9,10-octahydropyrimido[1,2-a]acoxane (7.8 g, 51.4 mmol, 2.3 equivalents) were added to the reaction mixture. The resulting mixture was then stirred at room temperature overnight. The mixture was then diluted with water (105 mL) and extracted with MTBE. The organic layers were combined, dried (Na₂SO₄), filtered, and concentrated under reduced pressure. The crude residue was purified by rapid column chromatography to yield 2,2'-di-tert-butyl5,5'-ethane-1,2-dimethylbis[(1R,4S,5S)-2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate] (6.65 g, 62% yield) as a colorless solid. LCMS (Method 3) RT = 2.25 min, m / z : [ESI + 481.40 (M+H) + .

[0327] Step B Synthesis of ethane-1,2-dimethylbis[(1R,4S,5S)-2-azabicyclo[2.1.1]hexane-5-carboxylate] hydrochloride A solution of 5 M HCl in isopropanol (48 mL, 17.5 equivalences) was added to a stirred solution of 2,2'-di-tert-butyl-5,5'-ethane-1,2-dimethylbis[(1R,4S,5S)-2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate] (6.65 g, 14 mmol) in isopropanol (150 mL). The reaction mixture was stirred overnight at room temperature. The reaction mixture was then poured into MTBE (220 mL) and stirred for 1 hour. The resulting precipitate was filtered, transferred to a round-bottom flask, azeotropically treated with MTBE and methanol, filtered, and air-dried to give ethane-1,2-dimethylbis[(1R,4S,5S)-2-azabicyclo[2.1.1]hexane-5-carboxylate] hydrochloride (2.9 g, 61% yield) as a grayish-white solid. LCMS (Method 3) RT = 5.23 min, m / z : [ESI + 281.1 (M+H) + . 1 H NMR (300 MHz, DMSO- d 6): δ 10.06 (br s, 2H), 8.94 (br s, 2H), 4.39 -4.36 (m, 2H), 4.32 - 4.23 (m, 4H), 3.29 - 3.21 (m, 6H), 3.11 - 3.06 (m, 2H),1.96 - 1.93 (m, 2H), 1.41 - 1.38 (m, 2H).

[0328] Example 38. Ethane-1,2-dimethylbis[(1R,4S,5S)-2-azabicyclo[2.1.1]hexane-5-carboxylate] hydrochloric acid Salt Step A Synthesis of 2,2'-di-tert-butyl5,5'-( 1S, 4S )-Cyclohexane-1,4-dimethylbis[(1R,4S,5S)-4-methyl-2- [Zazabicyclo[2.1.1]hexane-2,5-dicarboxylate] Towards( 1R, 4S, 5S )-2-(tert-butoxycarbonyl)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (9.0 g, 37 mmol, 2.2 equivalents) and trans-1,4-cyclohexanediol (2.0 g, 17 mmol) were added to a stirred solution in DCM (200 mL) with DMAP (2.1 g, 17 mmol, 1.0 equivalents) and 3-(((ethylimino)methylene)amino)- N , N2,2'-Dimethylpropyl-1-amine hydrochloride (7.8 g, 41 mmol, 2.4 equivalents). The reaction mixture was stirred at room temperature for 16 h. Subsequently, the reaction mixture was diluted with DCM (200 mL), washed with water (200 mL), 0.5 N HCl aqueous solution (200 mL), sodium bicarbonate aqueous solution (200 mL), and water (200 mL). The organic layer was dried (Na₂SO₄), filtered, concentrated under reduced pressure, and purified by rapid column chromatography, eluting with a mixture of ethyl acetate and heptane to give 2,2'-di-tert-butyl-5,5'-( 1S, 4S )-Cyclohexane-1,4-dimethylbis[(1R,4S,5S)-4-methyl-2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate] (7.4 g, 78% yield). LCMS (Method 3) RT = 2.68 min, m / z : [ESI + 463.40 (M-100+H) + .

[0329] Step B synthesis( 1S, 4S )-Cyclohexane-1,4-dimethylbis[(1R,4S,5S)-4-methyl-2-azabicyclo[2.1.1]hexane [Alkyl-5-carboxylate] hydrochloride To 2,2'-di-tert-butyl 5,5'-( 1S, 4S A solution of HCl aqueous solution in cyclopentyl methyl ether (1R,4S,5S)-4-methyl-2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate (7.4 g, 13.1 mmol) in cyclopentyl methyl ether (100 mL) was added to a stirred solution of cyclopentyl methyl ether (3.0 M, 100 mL). The solution was stirred at room temperature for 16 h. The reaction mixture was then concentrated under reduced pressure, and the residue was ground with pentane (100 mL). The precipitate was filtered, washed with pentane, and air-dried to give a grayish-white solid. 1S, 4S )-Cyclohexane-1,4-dimethylbis[(1R,4S,5S)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylate] hydrochloride (4.1 g, 72% yield). LCMS (Method 3) RT = 4.77 min, m / z : [ESI + 363.2 (M+H) + . 1 H NMR (400 MHz, DMSO- d6): δ 10.21 (br s, 2H), 8.66 (br s, 2H), 4.78 (s, 2H), 4.28 (s, 2H), 3.11 (m, 2H), 3.03 (m, 2H), 2.96 (s, 2H), 1.89 (t, J = 3.1 Hz, 4H), 1.75 (d, J = 8.0 Hz, 2H), 1.61 - 1.49 (m, 6H), 1.34 (s, 6H).

[0330] Example 39. Propane-1,3-dimethylbis[(1R,4S,5S)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylic acid Ester hydrochloride Step A Synthesis of 2,2'-di-tert-butyl5,5'-(propane-1,3-diyl)bis[(1R,4S,5S)-4-methyl-2-azabicyclo] [2.1.1] Hexane-2,5-dicarboxylate] Towards( 1R, 4S, 5S )-2-(tert-butoxycarbonyl)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (9.0 g, 37 mmol, 2.2 equivalents) and propane-1,3-diol (1.3 g, 17 mmol) were added to a stirred solution in DCM (200 mL) with DMAP (2.1 g, 17 mmol, 1.0 equivalents) and 3-(((ethylimino)methylene)amino)- N , N 1-Dimethylpropyl-1-amine hydrochloride (7.8 g, 41 mmol, 2.4 equivalents). The reaction mixture was stirred at room temperature for 80 h. Afterward, the reaction mixture was diluted with DCM (200 mL), water (100 mL), and HCl aqueous solution (0.5 mL). N The organic layer was washed with 100 mL of sodium bicarbonate aqueous solution and 50 mL of water. The organic layer was dried (Na₂SO₄), filtered, and concentrated under reduced pressure to give 2,2'-di-tert-butyl5,5'-(propane-1,3-diyl)bis[(1R,4S,5S)-4-methyl-2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate] (8.5 g, crude), which was used in the next step without further purification. LCMS (Method 3) RT = 2.46 min, m / z : [ESI + 523.40 (M+H) + .

[0331] Step B synthesisPropane-1,3-dimethylbis[(1R,4S,5S)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylate] hydrochloride A solution of HCl in cyclopentylmethyl ether (3.0 M, 100 mL) was added to a stirred solution of 2,2'-di-tert-butyl 5,5'-(propane-1,3-diyl)bis[(1R,4S,5S)-4-methyl-2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate] (8.5 g, 16 mmol) in cyclopentylmethyl ether (100 mL). The mixture was stirred at room temperature for 80 h. The reaction mixture was then concentrated under reduced pressure. The residue was dissolved in DCM and concentrated to dryness to give propane-1,3-diylbis[(1R,4S,5S)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylate] hydrochloride (5.5 g, 70% yield, in two steps) as a brown solid. LCMS (Method 3) RT = 4.68 min, m / z : [ESI + 323.2 (M+H) + . 1 H-NMR (400MHz, DMSO- d 6): δ 10.22 (br s, 2H), 8.85 (br s, 2H), 4.30 (s, 2H), 4.15 (m,2H), 4.10 (m, 2H), 3.09 (br s, 4H), 2.98 (s, 2H), 1.95 (t, J = 6.3 Hz, 2H), 1.76 (d, J = 8.0 Hz, 2H), 1.54 (d, J = 8.7 Hz, 2H), 1.34 (s, 6H).

[0332] Option 3 Add DMF (0.05 M) containing the corresponding 2-(tert-butoxycarbonyl)-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (II or V) (2.2 equivalents) to a stirred solution of CDI (2.2 equivalents) in DMF (0.05 M). After 2 h, add 1,8-diazabicyclo[5.4.0]undec-7-ene (2.3 equivalents) and the corresponding diol (1.0 equivalents), and stir the reaction mixture overnight at room temperature. Then, add MTBE and water, separate the phases, and dry the organic phase (Na2SO4), filter, and concentrate under reduced pressure. N-Boc deprotection steps (such as reaction with TFA, cyclopentylmethyl ether containing 3M HCl, or DCM containing formic acid) are used to prepare compounds of general formula (30). The mixture is stirred at room temperature for the required time. The volatiles are concentrated under reduced pressure, and the residue is dissolved in water. The aqueous layer is extracted with DCM, and the crude product is dried by centrifugal evaporation.

[0333] Example 40. 2,2-Dimethylpropane-1,3-dimethylbis[(1R,4S,5S)-2-azabicyclo[2.1.1]hexane-5- [Formate] Trifluoroacetate Step A synthesis 2,2'-Di-tert-butyl-5,5'-(2,2-dimethylpropane-1,3-diyl)bis[(1R,4S,5S)-2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate] Towards( 1R, 4S, 5S A solution of CDI (360 mg, 2.22 mmol, 2.2 equivalents) in DMF (44 mL) was added to a stirred solution of 2-(tert-butoxycarbonyl)-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (504 mg, 2.22 mmol, 2.2 equivalents) in DMF (44 mL). The mixture was stirred at room temperature for 0.5 h, followed by the addition of neopentyl glycol (105 mg, 1.0 mmol, 1.0 equivalents) and 1,8-diazabicyclo[5.4.0]undec-7-ene (353 mg, 2.32 mmol, 2.3 equivalents). The mixture was stirred at room temperature overnight. Subsequently, MTBE and water were added, and the mixture was stirred. The phases were separated, and the organic phase was dried (Na2SO4), filtered, and concentrated under reduced pressure. The residue was purified by rapid column chromatography and eluted with a mixture of cyclohexane and ethyl acetate to give 2,2'-di-tert-butyl-5,5'-(2,2-dimethylpropane-1,3-diyl)bis[(1R,4S,5S)-2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate] (158 mg, 30% yield) as a colorless solid. 1 H NMR (400 MHz, CDCl3) δ 4.59 - 4.53 (m, 2H), 3.95 - 3.48 (m, 4H), 3.32 - 3.25 (m, 2H), 3.04 (br s, 2H), 2.80 (br s, 2H), 1.82 - 1.77 (m, 2H), 1.45 (s, 18H), 1.35 - 1.30 (m, 2H), 0.93 - 0.88 (m, 8H).

[0334] Step B synthesis2,2-Dimethylpropane-1,3-dimethylbis[(1R,4S,5S)-2-azabicyclo[2.1.1]hexane-5-carboxylate] Trifluoroacetate 2,2'-Di-tert-butyl-5,5'-(2,2-dimethylpropane-1,3-diyl)bis[(1R,4S,5S)-2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate] (85 mg, 0.16 mmol) was dissolved in TFA (2 mL), and the mixture was stirred at room temperature for 0.25 h. The reaction mixture was then concentrated under reduced pressure. The residue was dissolved in water, the aqueous layer was washed with DCM, and the crude product was freeze-dried to give 2,2-dimethylpropane-1,3-diylbis[(1R,4S,5S)-2-azabicyclo[2.1.1]hexane-5-carboxylate] (93 mg, quantitative yield) as a colorless oil. LCMS (Method 4) RT = 0.87 min, m / z : [ESI + 323.1 (M+H) + . 1 H NMR (400 MHz, D2O) δ 4.46 (dd, J = 4.7, 2.6 Hz, 2H), 4.06 -3.90 (m, 4H), 3.48 (s, 4H), 3.30 (d, J = 6.7 Hz, 4H), 2.13 (d, J = 8.8 Hz, 2H), 1.45 (d, J = 9.5 Hz, 2H), 0.98 (s, 6H), no exchangeable protons were observed.

[0335] The following compounds were prepared according to scheme 3 of the above methods:

[0336] Example 51. 2-Azide-propane-1,3-dimethylbis[(1R,4S,5S)-2-azabicyclo[2.1.1]hexane-5-methyl [Acid Ester] Trifluoroacetate Step A synthesis 2,2'-Di-Tertiary Butyl 5,5'-(2-hydroxypropane-1,3-diyl)bis[(1R,4S,5S)-2-azabicyclo] [2.1.1] Hexane-2,5-dicarboxylate] Towards( 1R, 4S, 5SA solution of CDI (774 mg, 4.77 mmol, 2.2 equivalents) in DMF (95 mL) was added to a stirred solution of 2-(tert-butoxycarbonyl)-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (1083 mg, 4.77 mmol, 2.2 equivalents) in DMF (95 mL). The mixture was stirred at room temperature for 0.5 h, followed by the addition of propane-1,2,3-triol (200 mg, 2.17 mmol, 1 equivalent) and 1,8-diazabicyclo[5.4.0]undec-7-ene (760 mg, 4.99 mmol, 2.3 equivalents). The mixture was stirred at room temperature overnight. Subsequently, MTBE and water were added, and the mixture was stirred rapidly. The phases were separated, and the organic phase was dried (Na2SO4), filtered, and concentrated under reduced pressure. The residue was purified by rapid column chromatography, eluted with a mixture of cyclohexane and ethyl acetate, to give O2 as a colorless oil. 5 -[3-[(1R,4S)-2-tert-butoxycarbonyl-2-azabicyclo[2.1.1]hexane-5-carbonyl]oxy-2-hydroxy-propyl] O 2 -tert-butyl(1R,4S)-2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate (296 mg, 27% yield). 1 H NMR (400 MHz, CDCl3) δ 4.58 (s, 2H), 4.35 - 4.18 (m,1H), 4.05 - 4.01 (m, 2H), 3.62 - 3.57 (m, 3H), 3.36 - 3.18 (m, 3H), 3.05 (dd, J = 6.7, 3.0 Hz, 2H), 2.81 (dd, J = 7.4, 4.7 Hz, 2H), 1.81 - 1.77 (m, 2H), 1.46(d, J = 2.6 Hz, 18H), 1.38 - 1.31 (m, 2H).

[0337] Step B Synthesis of 5,5'-(2-azidopropane-1,3-diyl)2,2'-di-tert-butylbis[(1R,4S,5S)-2-azabis] Cyclohexane-2,5-dicarboxylate Triphenylphosphine (306 mg, 1.17 mmol, 2.0 equivalence) was added to a stirred solution of 2,2'-di-tert-butyl-5,5'-(2-hydroxypropane-1,3-diyl)bis[(1R,4S,5S)-2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate] (296 mg, 0.58 mmol) in THF (7 mL), followed by diisopropyl azodicarbonate (252 µL, 1.17 mmol, 2.0 equivalence) and diphenylphosphoazide (85 µL, 0.39 mmol, 2.0 equivalence). The resulting mixture was stirred overnight at room temperature. The reaction mixture was then concentrated under reduced pressure, filtered, and the solids were washed with ethyl acetate. The filtrate was concentrated under reduced pressure and purified by rapid column chromatography, eluted with a mixture of ethyl acetate and cyclohexane to give 5,5'-(2-azidopropane-1,3-diyl)2,2'-di-tert-butylbis[(1R,4S,5S)-2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate] (209 mg, 67% yield) as a colorless oil. 1 H NMR (CDCl3, 400 MHz) δ 4.57 (s, 2H), 4.21 - 4.11 (m, 3H), 4.03 (dd, J = 11.6, 6.5 Hz, 1H), 3.89 - 3.76 (m, 1H), 3.55 (s, 2H), 3.27 (d, J = 8.8 Hz, 2H), 3.17 - 2.97 (m, 2H), 2.90 - 2.75 (m, 2H), 1.83 - 1.81(m, 2H), 1.46 (d, J = 0.5 Hz, 18H), 1.34 (dd, J = 7.6, 1.1 Hz, 2H).

[0338] Step C Synthesis of 2-azidopropane-1,3-dimethylbis[(1R,4S,5S)-2-azabicyclo[2.1.1]hexane-5-carboxylic acid [Ester] Trifluoroacetate 5,5'-(2-azidopropane-1,3-diyl)2,2'-di-tert-butylbis[(1R,4S,5S)-2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate] (100 mg, 0.19 mmol) was dissolved in TFA (2.5 mL), and the mixture was stirred at room temperature for 0.25 h. The reaction mixture was then concentrated under reduced pressure, and the residue was dissolved in water. The aqueous layer was washed with DCM, and the crude aqueous layer was freeze-dried to give 2-azidopropane-1,3-diylbis[(1R,4S,5S)-2-azabicyclo[2.1.1]hexane-5-carboxylate] trifluoroacetate (100 mg, 95% yield) as a colorless oil. LCMS (Method 4) RT = 1.13 min, m / z : [ESI + 335 (M+H) + . 1 ¹H NMR (D₂O, 400 MHz) δ 4.49 - 4.46 (m, 2H), 4.42 - 4.33 (m, 2H), 4.30 - 4.22 (m, 2H), 4.19 - 4.15 (m, 1H), 3.56 - 3.45 (m, 4H), 3.36 - 3.26 (m, 4H), 2.20 - 2.09 (m, 2H), 1.49 - 1.42 (m, 2H). No exchangeable protons were observed.

[0339] Example 52. 2R, 3R )-Butane-2,3-dimethylbis[(1R,4S,5S)-4-methyl-2-azabicyclo[2.1.1]hexane [Alkyl-5-carboxylate] hydrochloride Step A Synthesis of 2-(tert-butyl)5-(( 2R, 3R )-3-hydroxybutane-2-yl) ( 1R, 4S, 5S )-4-methyl-2-azabi Cyclo[2.1.1]hexane-2,5-dicarboxylate Towards( 1R, 4S, 5S A solution of CDI (139 mg, 0.85 mmol, 2.2 equivalents) in DMF (17 mL) was added to a stirred solution of 2-(tert-butoxycarbonyl)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (206 mg, 0.85 mmol, 2.2 equivalents) in DMF (17 mL). The mixture was stirred at room temperature for 0.5 h. Afterward, ( 2R, 3R2,3-butanediol (35 mg, 0.39 mmol, 1 equivalent) and 1,8-diazabicyclo[5.4.0]undec-7-ene (136 mg, 0.89 mmol, 2.3 equivalents) were added. The mixture was stirred overnight at room temperature. Subsequently, MTBE and water were added to the mixture and stirred, the phases were separated, and the organic phase was dried (Na2SO4), filtered, and concentrated under reduced pressure. The residue was purified by rapid column chromatography, eluting with a mixture of cyclohexane and ethyl acetate to give 2-(tert-butyl)-5-(( 2R, 3R )-3-hydroxybutane-2-yl)( 1R, 4S, 5S 4-methyl-2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate (122 mg, quantitative yield).

[0340] Step B synthesis 5,5'-(2 R ,3 R )-Butane-2,3-diyl-2,2'-di-tert-butylbis[(1 R 4 S 5 S )-4-methyl-2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate] Towards( 1R, 4S, 5S A solution of CDI (114 mg, 0.70 mmol, 2.2 equivalents) in DMF (14 mL) was added to a stirred solution of 2-(tert-butoxycarbonyl)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (169 mg, 0.70 mmol, 2.2 equivalents) in DMF (14 mL). The mixture was stirred at room temperature for 0.5 h. Afterward, 2-(tert-butyl)-5-(( 2R, 3R )-3-hydroxybutane-2-yl) ( 1R, 4S, 5S 1,8-Diazabicyclo[2.1.1]hexane-2,5-dicarboxylate (100 mg, 0.32 mmol, 1 equivalent) and 1,8-diazabicyclo[5.4.0]undec-7-ene (112 mg, 0.73 mmol, 2.3 equivalent). The mixture was stirred overnight at room temperature. MTBE and water were then added to the mixture and stirred, the phases were separated, and the organic phase was dried (Na₂SO₄), filtered, and concentrated under reduced pressure. The residue was purified by rapid column chromatography, eluting with a mixture of cyclohexane and ethyl acetate to give 5,5'-(2-dicarboxylate) as a yellow oil. R ,3 R )-Butane-2,3-diyl-2,2'-di-tert-butylbis[(1 R 4 S 5S [4-methyl-2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate] (116 mg, 68% yield). 1 HNMR (400 MHz, CDCl3) δ 4.95 - 4.92 (m, 2H), 4.62 - 4.41 (m, 2H), 3.36 (s,2H), 3.05 (d, J = 8.6 Hz, 2H), 2.47 (d, J = 8.4 Hz, 2H), 1.57 - 1.54 (m, 2H), 1.45 (s, 18H), 1.41 (dd, J = 7.3, 1.0 Hz, 2H), 1.37 (s, 6H), 1.19 - 1.11 (m, 6H).

[0341] Step C Synthesis of (2R,3R)-butane-2,3-dimethylbis[(1R,4S,5S)-4-methyl-2-azabicyclo[2.1.1]hexane- 5-Carbamate] hydrochloride Will 5,5'-(2 R ,3 R )-Butane-2,3-diyl-2,2'-di-tert-butylbis[(1 R 4 S 5 S )-4-methyl-2-aza Bicyclo[2.1.1]hexane-2,5-dicarboxylate] (111 mg, 0.21 mmol, 1.0 equivalent) and CPME containing HCl (3 M, 0.83 mL) were stirred overnight at room temperature. The solvent was then concentrated under reduced pressure to give (2R,3R)-butane-2,3-dimethylbis[(1R,4S,5S)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylate] hydrochloride (84 mg, 99% yield) as a grayish-white oil. LCMS (Method 4) RT = 2.58 min, m / z : [ESI + 337.1 (M+H) + . 1 H NMR (400 MHz, D2O) δ5.12 - 5.05 (m, 2H), 4.40 (t, J = 1.6 Hz, 2H), 3.34 (dd, J = 10.2, 1.3 Hz, 2H), 3.28 (dd, J = 10.3, 2.2 Hz, 2H), 3.08 - 3.05 (m, 2H), 1.96 (dt, J = 9.4, 2.0 Hz, 2H), 1.59 (dd, J= 9.3, 1.2 Hz, 2H), 1.47 (s, 6H), 1.29 - 1.24 (m, 6H), No exchangeable protons were observed.

[0342] Example 53. 1R, 4R )-Cyclohexane-1,4-dimethylbis[(1S,4R,5R)-4-methyl-2-azabicyclo[2.1.1] [Hexane-5-carboxylate] hydrochloride Step A Synthesis of 2,2'-di-tert-butyl 5,5'-(( 1R, 4R )-Cyclohexane-1,4-diyl)bis[(1S,4R,5R)-4-methyl- 2-Zazabicyclo[2.1.1]hexane-2,5-dicarboxylate] Towards( 1S, 4R, 5R )-2-(tert-butoxycarbonyl)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (1.0 g, 4.14 mmol) and trans-1,4-cyclohexanediol (0.48 g, 4.14 mmol, 1.04 equivalents) were added to a stirred solution of DMF (22 mL) with DMAP (0.51 g, 4.14 mmol, 1.0 equivalents) and 3-(((ethylimino)methylene)amino)- N , N -Dimethylpropyl-1-amine hydrochloride (1.9 g, 9.9 mmol, 2.4 equivalents). The reaction mixture was stirred at room temperature for 16 h. Subsequently, the reaction mixture was diluted with DCM (35 mL) and washed with water (35 mL), 0.5 N HCl aqueous solution (35 mL), and sodium bicarbonate aqueous solution (35 mL), and washed again with water (35 mL). The organic layer was dried (Na₂SO₄), filtered, concentrated under reduced pressure, and purified by rapid column chromatography, eluting with a mixture of ethyl acetate and heptane to give a grayish-white solid. 2,2'-Second Uncle Ding Base 5,5'-(( 1R, 4R )-cyclohexane-1,4-diyl)bis[(1S,4R,5R)-4-methyl-2-azabicyclo[2.1.1]hexane-2, [5-Dicarboxylate] (0.53 g, 23% yield). LCMS (Method 3) RT = 2.57 min, m / z : [ESI + 463.40 (M-100+H) + .

[0343] Step B synthesis ( 1R, 4R )-Cyclohexane-1,4-dimethylbis[(1S,4R,5R)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylate] hydrochloride To 2,2'-di-tert-butyl 5,5'-(( 1R, 4RA solution of HCl in cyclopentylmethyl ether (3.0 M, 4.6 mL) was added to a stirred solution of cyclopentylmethyl ether (8 mL). The solution was stirred at room temperature for 16 h. Then, another cyclopentylmethyl ether containing HCl (3.0 M, 1 mL) was added. The reaction mixture was stirred for another 12 h. The reaction mixture was then concentrated under reduced pressure, and the residue was diluted with pentane. The resulting precipitate was filtered, washed with pentane, and air-dried to give a grayish-white solid (…). 1R, 4R )-Cyclohexane-1,4-dimethylbis[(1S,4R,5R)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylate] hydrochloride (0.22 g, 55% yield). LCMS (Method 3) RT = 4.64 min, m / z : [ESI + 363.2 (M+H) + . 1 H-NMR (400 MHz, CD3OD) δ 4.93 - 4.91 (m, 2H), 4.33 (t, J = 1.7 Hz, 2H), 3.24 - 3.22 (m, 4H), 2.98 - 2.96 (m, 2H), 2.03 - 1.99 (m, 4H), 1.93 - 1.91 (m, 2H), 1.63 - 1.61 (m, 4H), 1.57 - 1.54 (m, 2H), 1.44 (s, 6H), No exchangeable protons were observed.

[0344] Example 54. Propane-1,3-dimethylbis[(1S,4R,5R)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylic acid Ester hydrochloride Step A synthesis 2,2'-Di-tert-butyl5,5'-(propane-1,3-diyl)bis[(1S,4R,5R)-4-methyl-2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate] Towards( 1S, 4R, 5R)-2-(tert-butoxycarbonyl)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (1.0 g, 4.14 mmol) and propane-1,3-diol (0.32 g, 4.14 mmol, 1 equivalent) were added to a stirred solution in DCM (22 mL) with DMAP (0.51 g, 4.14 mmol, 1.0 equivalent) and 3-(((ethylimino)methylene)amino)- N , N 1-Dimethylpropyl-1-amine hydrochloride (1.9 g, 9.9 mmol, 2.4 equivalents). The reaction mixture was stirred at room temperature for 16 h. Subsequently, the reaction mixture was diluted with DCM (35 mL) and with water (35 mL) and HCl aqueous solution (0.5 mL). N The organic layer was washed with 35 mL of sodium bicarbonate aqueous solution and 35 mL of water. The organic layer was dried (Na₂SO₄), filtered, and concentrated under reduced pressure to give 2,2'-di-tert-butyl-5,5'-(propane-1,3-diyl)bis[(1S,4R,5R)-4-methyl-2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate] (0.8 g, 36% yield) as an amber oil. LCMS (Method 3) RT = 2.43 min, m / z : [ESI + 523.40 (M+H) + .

[0345] Step B synthesis Propane-1,3-dimethylbis[(1S,4R,5R)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylate] hydrochloride A solution of HCl in cyclopentylmethyl ether (3.0 M, 7.5 mL) was added to a stirred solution of 2,2'-di-tert-butyl 5,5'-(propane-1,3-diyl)bis[(1S,4R,5R)-4-methyl-2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate] (0.8 g, 1.5 mmol) in cyclopentylmethyl ether (8 mL). The mixture was stirred at room temperature for 16 h. The reaction mixture was then concentrated under reduced pressure and purified by HPLC to give propane-1,3-diylbis[(1S,4R,5R)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylate] hydrochloride (0.5 g, 71% yield) as a grayish-white solid. LCMS (Method 3) RT = 4.57 min, m / z : [ESI + 323.2 (M+H) + .1 1H-NMR (400 MHz, CD3OD): δ 4.31 - 4.26 (m, 2H), 4.24 - 4.21 (m, 2H), 4.17 - 4.11 (m, 2H), 3.28 - 3.21 (m, 4H), 2.95 - 2.93 (m, 2H), 2.02 - 1.95 (m, 2H), 1.8 - 1.85 (m, 2H), 1.54 - 1.52 (m, 2H), 1.40 (s, 6H). No exchangeable protons were observed.

[0346] Example 55.(1) S 4 S )-4-{[(1 R 4 S 5 S )-2-azabicyclo[2.1.1]hexane-5-carbonyl]oxy}cyclohexane Base (1) R 4 S 5 S 4-Methyl-2-azabicyclo[2.1.1]hexane-5-carboxylate hydrochloride Step A synthesis( 1r,4r 4-((tert-butyldimethylsilyl)oxy)cyclohexane-1-ol To a stirred solution of trans-cyclohexanediol (25 g, 0.22 mol, 1.0 equivalent) in THF (250 mL), tert-butyldimethylchlorosilane (32 g, 0.22 mol, 1.0 equivalent) and imidazole (15 g, 0.22 mol, 1 equivalent) were added. The reaction mixture was stirred overnight at room temperature. The mixture was then filtered, and the filtrate was concentrated under reduced pressure. The residue was ground with chloroform (200 mL), and the resulting solid was filtered and purified by rapid column chromatography, eluting with a mixture of ethyl acetate and heptane to give a colorless syrup. 1r,4r )-4-((tert-butyldimethylsilyl)oxy)cyclohexane-1-ol (12 g, 48% yield).

[0347] Step B Synthesis of 2-tert-butyl-5-[(1 r 4 S )-4-{[tert-butyldi(methyl)silyl]oxy}cyclohexyl](1 R 4 S 5 S )- 2-Zazabicyclo[2.1.1]hexane-2,5-dicarboxylate Add ( ) to a solution of (1R,4S,5S)-2-[(tert-butoxy)carbonyl]-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (6.5 g, 29 mmol, 1.1 equivalents) in DCM (200 mL) 1r,4r4-((tert-butyldimethylsilyl)oxy)cyclohexane-1-ol (6.0 g, 26 mmol, 1 equivalent), EDC (5.5 g, 29 mmol, 1.1 equivalent), and DMAP (1.6 g, 13 mmol, 0.5 equivalent). The reaction mixture was stirred at room temperature for 16 h. The mixture was then washed successively with water (200 mL), 0.5 NHCl aqueous solution (200 mL), sodium bicarbonate aqueous solution (200 mL), and water (100 mL). The organic phase was dried (Na₂SO₄), filtered, concentrated under reduced pressure, and purified by rapid column chromatography, eluting with a mixture of ethyl acetate and heptane to give a grayish-white solid. 2-tert-butyl-5-[(1 r 4 S )-4-{[tert-butyldi(methyl)silyl]oxy}cyclohexyl](1 R 4 S 5 S )-2-Nitrogen Heterobicyclo[2.1.1]hexane-2,5-dicarboxylate (6.2 g, 54% yield). 1 H-NMR (400 MHz, CDCl3): δ 4.71- 4.55 (m, 2H), 3.68 - 3.47 (m, 2H), 3.25 - 3.21 (m, 1H), 3.01 - 2.97 (m,1H), 2.73 - 2.71 (m, 1H), 1.88 - 1.59 (m, 8H), 1.43 - 1.41 (m, 18H), 1.39 -1.35 (m, 1H), 1.28 - 1.25 (m, 1H), 0.88 (s, 6H).

[0348] Step C Synthesis of 2-tert-butyl-5-[( 1r,4S )-4-hydroxycyclohexyl]( 1R, 4S, 5S )-2-azabicyclo[2.1.1]hexane- 2,5-Dicarboxylate To 2-tert-butyl5-[(1 r 4 S )-4-{[tert-butyldi(methyl)silyl]oxy}cyclohexyl](1 R 4 S 5 S 2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate (4.3 g, 10 mmol) was added to a stirred solution of 2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate in THF (100 mL) with tetrabutylammonium fluoride (1.0 M in THF, 15 mL, 15 mmol, 1.5 equivalent). The reaction mixture was stirred at 40 °C for 16 h. Afterward, the mixture was poured into water (200 mL) and extracted with MTBE. The organic phase was washed with an aqueous solution of sodium bicarbonate (100 mL), followed by washing with water (100 mL), dried (Na₂SO₄), filtered, and concentrated under reduced pressure to give 2-tert-butyl-5-[( 1r, 4S )-4-hydroxycyclohexyl]( 1R, 4S, 5S2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate (3.4 g, quantitative), which was used directly without further purification.

[0349] Step D Synthesize 5-[(1 S 4 S )-4-{[(1 R 4 S 5 S )-2-(tert-butoxycarbonyl)-2-azabicyclo[2.1.1]hexane- 5-carbonyl]oxy]cyclohexyl]2-tert-butyl(1 R 4 S 5 S )-4-methyl-2-azabicyclo[2.1.1]hexane-2,5-dicarboxylic acid ester To 2-tert-butyl5-[( 1r,4S )-4-hydroxycyclohexyl]( 1R, 4S, 5S )-2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate (3.4 g, 10 mmol) was added to a stirred solution in DCM (100 mL). 1R, 4S, 5S 2-(tert-butoxycarbonyl)-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylic acid (2.8 g, 11 mmol, 1.1 equivalents) was added, followed by the addition of EDCI (2.2 g, 11 mmol, 1.1 equivalents) and DMAP (0.64 g, 5.2 mmol, 0.5 equivalents). The reaction mixture was stirred at room temperature for 16 h. The mixture was then washed successively with water (100 mL), 0.5 N HCl aqueous solution (50 mL), sodium bicarbonate aqueous solution (50 mL), and water (50 mL). The organic phase was dried (Na2SO4), concentrated under reduced pressure, and purified by rapid column chromatography, eluting with a mixture of ethyl acetate and heptane to give 5-[(1 S 4 S )-4-{[(1 R 4 S 5 S )-2-(tert-butoxycarbonyl)-2-azabicyclo[2.1.1]hexane-5-carbonyl]oxy}cyclohexyl]2-tert-butyl(1 R 4 S 5 S 4-Methyl-2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate (3.0 g, 52% yield). LCMS (Method 3) RT = 6.71 min, m / z : [ESI + 449.4 (M-100+H) + .

[0350] Step E Synthesis (1) S 4 S )-4-{[(1 R 4 S 5 S )-2-azabicyclo[2.1.1]hexane-5-carbonyl]oxy}cyclohexyl (1 R 4 S 5 S )-4-methyl-2-azabicyclo[2.1.1]hexane-5-carboxylate To 5-[(1 S 4 S )-4-{[(1 R 4S 5 S )-2-(tert-butoxycarbonyl)-2-azabicyclo[2.1.1]hexane-5-carbonyl]oxy}cyclohexyl]2-tert-butyl(1 R 4 S 5 S A solution of HCl in cyclopentylmethyl ether (3.0 M; 40 mL, 120 mmol, 20 equivalents) was added to a stirred solution of 4-methyl-2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate (3.0 g, 5.5 mmol) in diethyl ether (60 mL). The reaction mixture was stirred at room temperature for 16 h. Then, another solution of HCl in cyclopentylmethyl ether (10 mL) was added, and the reaction mixture was stirred for another 6 h. The mixture was then concentrated under reduced pressure to a solvent volume of ~40 mL, followed by the addition of pentane (40 mL). The resulting solid was separated by filtration and dried under reduced pressure to give a grayish-white solid (1... S 4 S )-4-{[(1 R 4 S 5 S )-2-azabicyclo[2.1.1]hexane-5-carbonyl]oxy}cyclohexyl(1 R 4 S 5 S 4-Methyl-2-azabicyclo[2.1.1]hexane-5-carboxylate (1.8 g, 78% yield). LCMS (Method 3) RT = 4.48 min, m / z : [ESI + 349.2 (M+H) + . 1 H NMR (400 MHz, DMSO- d 6) δ 10.36 -10.28 (m, 2H), 8.78 - 8.71 (m, 2H), 4.73 (br s, 2H), 4.66 (br s, 2H), 4.34(d, J = 5.9 Hz, 1H), 4.27 (s, 1H), 3.26 - 3.23 (m, 2H), 3.17 - 3.15 (m, 1H), 3.12 - 2.98 (m, 3H), 2.95 (s, 1H), 1.89 - 1.83 (m, 4H), 1.74 (d, J = 7.7 Hz,1H), 1.61 - 1.46 (m, 4H), 1.42 (d, J= 8.4 Hz, 1H), 1.33 (s, 3H).

[0351] Example 56: 3-[(cyclopentanecarbonyl)oxy]propyl 2-azabicyclo[2.1.1]hexane-5-carboxylic acid trifluoroacetate Step A Synthesis of 2-tert-butyl5-{3-[(cyclopentanecarbonyl)oxy]propyl}2-azabicyclo[2.1.1]hexane-2,5-di Formate To a stirred solution of 2-tert-butyl5-(3-hydroxypropyl)2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate (460 mg, 1.62 mmol) and cyclopentanecarboxylic acid (185 mg, 1.62 mmol, 1.0 equivalence) in anhydrous DCM at 0 °C, DCC (401 mg, 1.95 mmol, 1.2 equivalence) and DMAP (10 mg, 0.08 mmol) were slowly added. The mixture was stirred overnight at room temperature. The resulting precipitate was then filtered. The organic phase was washed with water, dried (Na2SO4), filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC to give 2-tert-butyl5-{3-[(cyclopentanecarbonyl)oxy]propyl}2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate (242 mg, 43% yield). LCMS (Method 1) RT = 1.28 min, m / z : [ESI + 282.2 (M-100+H) + .

[0352] Step B Synthesis of 3-[(cyclopentanecarbonyl)oxy]propyl trifluoroacetate of 2-azabicyclo[2.1.1]hexane-5-carboxylic acid TFA (72 mg, 1.2 mmol) was added to a stirred solution of 2-tert-butyl-5-{3-[(cyclopentanecarbonyl)oxy]propyl}2-azabicyclo[2.1.1]hexane-2,5-dicarboxylate (242 mg, 0.64 mmol) in DCM at 0 °C. The solution was then stirred overnight at room temperature. The mixture was subsequently concentrated under reduced pressure. The residue was purified by preparative HPLC to give 2-azabicyclo[2.1.1]hexane-5-carboxylic acid 3-[(cyclopentanecarbonyl)oxy]propyl trifluoroacetate (54 mg, 22% yield) as a yellow oil. LCMS (Method 1) RT = 0.92 min, m / z : [ESI + 282.19 (M+H) + . 1 H NMR (400MHz, DMSO- d6) δ 9.40 (s, 1H), 8.62 (s, 1H), 4.38 (d, J = 6.0 Hz, 1H), 4.21 -3.97 (m, 4H), 3.25 - 3.20 (m, 3H), 3.10 (dt, J = 6.1, 3.2 Hz, 1H), 2.73 - 2.70(m, 1H), 2.02 - 1.73 (m, 6H), 1.73 - 1.44 (m, 5H), 1.27 (d, J = 8.5 Hz, 1H).

[0353] Example 57: Bioactivity Mitotracker analysis (MTR) On day 1, 25,000 U937 cells per well were seeded into 96-well plates containing either the compound of interest or a DMSO control, and incubated at 37°C and 5% CO2 for 48 h. On day 3, Mitotracker Red CMXRos was added to each well at a final concentration of 200 nM, and the plates were incubated at 37°C for 1 h in a CO2 incubator. After incubation, the fluorescence of the dye was acquired on a Quantum laser using the following parameters: yellow laser gain of 300 and PE-Cy5-H emission channel.

[0354] Data analysis included median PC5.5 fluorescence of live cells, percentage of live cells, and cell count / µL. All data points were normalized relative to the medium CTL.

[0355] Reagents: RPMI (Gibco, catalog number 61870-010), FBS (Gibco, Brazil origin, catalog number 10270-106), Pen-Strep (Gibco™ 15140122), MitoTracker RedCMXRos (ThermoFisherScientific, catalog number M7512) The results are presented as the minimum effective compound concentration with a 105% increase in MTR signal relative to the control (A = ≥ 30 µM; B = between 29.9 µM and 15 µM; C = between 14.9 µM and 5 µM; D = ≤ 4.9 µM).

[0356] Fluorescently labeled mitochondrial protein Cox8 MTS reporter assay The fully described prosequence of cytochrome oxidase subunit 8 (COX8) was fused to the N-terminus of a reporter fluorescent protein (codon-optimized versions of Dasher, Twinkle, and Rudolph (ATUM Bio)) to enable the reporter fluorescent protein to be transported into mitochondria. This construct was cloned into a plasmid under the constitutive promoter of EF1α and co-expressed with a puromycin resistance gene for mammalian cell selection. K562 cells were transfected and selected to obtain stable expression of Cox8-dasherGFP. On day 1, 25,000 K562 Cox8-dasherGFP cell plates were seeded into each well of a 96-well plate with either the compound of interest or a DMSO control and incubated at 37°C and 5% CO2 for 48 h. On day 3, fluorescence of the reporter DasherGFP located in the mitochondria was collected using a NovoCyte Quanteon flow cytometer. Data analysis included median fluorescence of viable cells, percentage of viable cells, and number of cells per μL. All data points were normalized relative to the median cytokine (CTL). The results are presented as the percentage increase in MTS signal relative to the control at a given test concentration (A = 101% - 104.9%; B = 105% - 108.9%; C = 109% - 112.9%; D = ≥ 113%).

[0357] High-resolution breathalyzer (HRR) Mitochondrial oxygen dynamics were measured using the high-resolution respiration assay OROBOROS Oxygraph-2k. On day 1, 235,000 U937 cells per mL (grown in RPMI / 10% FBS / PS) plates were seeded in RPMI / 10% FBS / PS containing either the test compound or DMSO control and incubated at 37°C and 5% CO2 for 48 h.

[0358] 1. Before starting the experiment, warm the culture medium (RPMI / 10% FBS / PS) to room temperature. 2. Remove 70% of the ethanol from the stopper and chamber, and rinse three times with HCl water. 3. Add 2.5 mL of culture medium (RPMI / 10% FBS / PS) to each chamber to flush the stopper. Aspirate the chamber and add 2.5 mL of freshly prepared culture medium. 4. Remove the culture medium from the chamber and replace it with 2.5 mL of culture medium (RPMI / 10% FBS / PS). Perform air calibration on the instrument and save the file for calibration during sample runs. 5. Remove the culture medium from the chamber and replace it with 2.5 mL of cell-containing medium (RPMI / 10% FBS / PS). The cells have been treated with the test compound or DMSO control and counted before the experiment. 6. Add 5 μL of 20 μM oligomycin solution and take readings until stable. 7. Add 5 μL of 250 μM CCCP solution until the maximum reading is reached. For U937 cells, the maximum reading was reached after the second injection of 5 μL of 250 μM CCCP. Perform a third injection to confirm that the reading is lower than the previous reading. 8. Add 4 μL of 1 mM rotenone solution, followed by 3 μL of 2.5 mM antimycin A solution to inhibit mitochondrial respiration. 9. After the experiment, save the file, and wash the chamber three times with 100% ethanol, rinse three times with 70% ethanol, and rinse three times with HCl water. 10. For analysis, use the DatLab program to select the longest stable reading range for quantification. The results are presented as the percentage increase in ATP-related respiration caused by the compound at the given test concentration (A = 101% - 104.9%; B = 105% - 108.9%; C = 109% - 112.9%; D = ≥ 113%).

[0359] Data table.

[0360] Example 57: Pharmaceutical Preparations (i) Tablet formulations A tablet composition containing the compound as defined in any one of embodiments 1.1 to 1.122 can be prepared by mixing 50 mg of the compound with 197 mg of lactose (BP) as a diluent and 3 mg of magnesium stearate as a lubricant and compressing in a known manner to form a tablet.

[0361] (ii) Capsule formulations A capsule formulation was prepared by mixing 100 mg of a compound as defined in any one of embodiments 1.1 to 1.122 with 100 mg of lactose and filling the resulting mixture into a standard opaque hard gelatin capsule.

[0362] (iii) Injectable formulation I A parenteral composition for injection can be prepared by dissolving the compound as defined in any one of embodiments 1.1 to 1.122 in water containing 10% propylene glycol to obtain an active compound concentration of 1.5% by weight. The solution is then sterilized by filtration, filled into ampoules, and sealed.

[0363] (iv) Injectable formulation II The parenteral composition for injection is prepared by dissolving a compound (2 mg / mL) as defined in any one of embodiments 1.1 to 1.122 and mannitol (50 mg / mL) in water, sterilely filtering the solution and filling it into a sealable 1 ml vial or ampoule.

[0364] v) Injectable formulation III A formulation for intravenous (iv) delivery by injection or infusion can be prepared by dissolving a compound (e.g., in salt form) as defined in any one of embodiments 1.1 to 1.122 in water at a concentration of 20 mg / ml. The vial is then sealed and sterilized by autoclaving.

[0365] vi) Injectable formulations IV A formulation for intravenous delivery by injection or infusion can be prepared by dissolving a compound (e.g., in salt form) as defined in any one of embodiments 1.1 to 1.122 at a concentration of 20 mg / ml in water containing a buffer solution (e.g., 0.2 M acetate, pH 4.6). The vial is then sealed and sterilized by autoclaving.

[0366] (vii) Subcutaneous injection preparations A composition for subcutaneous administration is prepared by mixing a compound as defined in any one of embodiments 1.1 to 1.122 with pharmaceutical-grade corn oil to obtain a concentration of 5 mg / ml. The composition is sterilized and filled into a suitable container.

[0367] viii) Lyophilized preparations Aliquots of the compound prepared as defined in any one of embodiments 1.1 to 1.122 were placed in 50 ml vials and lyophilized. During lyophilization, the composition was frozen at -45°C using a one-step freezing protocol. The temperature was raised to -10°C for annealing, then lowered to -45°C for freezing, followed by primary drying at +25°C for approximately 3400 minutes, and then secondary drying, during which the temperature was gradually increased to 50°C. The pressure during both primary and secondary drying was set to 80 mTorr.

[0368] equivalent The foregoing examples are presented for illustrative purposes and should not be construed as imposing any limitation on the scope of the invention. It will be readily apparent that many modifications and alterations can be made to the specific embodiments of the invention described above and illustrated in the examples without departing from the fundamental principles of the invention. This application is intended to cover all such modifications and alterations.

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Claims

1. A compound of formula (1): X 1 -L-X 2 (1) or its salts or tautomers; wherein X 1 and X 2 Choose independently from equations (2A), (2B), (2C), and (2D): (2A) (2B) (2C) (2D) in Indicates the connection point with group L, provided that X 1 and X 2 At least one of them has equation (2A). R 2 Selected from hydrogen, C, optionally substituted with hydroxyl groups 1-4 The hydrocarbon group and a 5-6 membered cyclic aromatic group or a 3-6 membered cyclic non-aromatic group, wherein the cyclic aromatic group and the cyclic non-aromatic group are optionally substituted by one or more substituents selected from: halogen, C 1-4 Alkyl, hydroxyl, C 1-4 Alkoxy and C 1-4 Alkyl group; and R 3 and R 4 One of them is selected from hydrogen and C, which is optionally substituted with a 5-6 membered cyclic aromatic group. 1-4 The hydrocarbon group, wherein the cyclic aromatic group is optionally substituted by one or more substituents selected from: halogen, cyano, hydroxyl, C 1-4 Alkyl and C 1-4 Alkoxy; and R 3 and R 4 The other one is hydrogen; L is an optionally substituted cyclic or acyclic linker group, which in X 1 With X 2 The chain length is 2 to 8 carbon atoms, wherein one of the carbon atoms may optionally be replaced by a heteroatom selected from O, N, S and their oxidized forms, provided that the heteroatom (if present) is in relation to X. 1 and X 2 There are always at least two carbon atoms between each of them.

2. The compound according to claim 1, wherein L is an optionally substituted cyclic or acyclic linking group, which in X 1 With X 2 The chain length is 2 to 6 carbon atoms, wherein one of the carbon atoms may optionally be replaced by a heteroatom or heteroatom group selected from O, N, S, S(O) and SO2, provided that the heteroatom (if present) is in contact with X. 1 and X 2 There are always at least two carbon atoms between each of them.

3. The compound according to claim 1, wherein L is an optionally substituted cyclic or acyclic linking group, which in X 1 With X 2 The chain length is 2 to 5 carbon atoms, wherein one of the carbon atoms may optionally be replaced by a heteroatom selected from O, N, and S, provided that the heteroatom (if present) is in relation to X. 1 and X 2 There are always at least two carbon atoms between each of them.

4. The compound according to claim 1, wherein L has the formula A 1 -BA 2 ,in: A 1 and A 2 C-type bonds independently selected from the bond and optionally substituted with one or more hydroxyl groups or halogens. 1-4 Hydrocarbon linkers; B is selected from the bond, C(R) b )2、N(R b ), O, S, S(O), SO2, 3 to 6-membered cyclic non-aromatic groups and 5 or 6-membered cyclic aromatic groups, wherein the cyclic non-aromatic groups and the cyclic aromatic groups are optionally surrounded by one or more groups R b Replace; and R b Selected from hydrogen, hydroxyl, C 1-4 hydrocarbon groups and -OC 1-4 Hydrocarbon groups, wherein each hydrocarbon group may optionally be replaced by a hydroxyl group, a halogen group, or a cyano group; The condition is A 1 -BA 2 The chain length between X1 and X2 is at least 2 atoms.

5. The compound according to claim 4, wherein A 1 and A 2 same.

6. The compound according to claim 4, wherein: A 1 and A 2 Selected independently from C 1-4 Hydrocarbon linkers; B is selected from bonds, 3- to 6-membered carbon ring non-aromatic groups, and 6-membered carbon ring aromatic groups.

7. The compound according to any one of claims 1 to 6, wherein A 1 and A 2 Both are bonds; or both are CH2 linkers; and B is a 4- to 6-membered carbon ring non-aromatic group or a 6-membered carbon ring aromatic group.

8. The compound according to any one of claims 1 to 6, wherein L is C 2-5 Hydrocarbon linkers (e.g., those with the formula (CH2)) n (where n is 2 to 5).

9. The compound according to any one of claims 1 to 8, wherein X 1 and X 2 Both have equation (2A).

10. The compound according to any one of claims 4 to 9, wherein A 1 and A 2 Same, and X 1 and X 2 Both have equation (2A).

11. The compound according to any one of claims 1 to 10, wherein R 2 Selected from hydrogen, methyl, and phenyl.

12. The compound according to any one of claims 1 to 11, wherein R 3 and R 4 Both are hydrogen.

13. The compound according to any one of claims 1 to 12, wherein X 1 and / or X 2 It has the following formula (AE): (AE) in Indicates the connection point with group L.

14. A compound selected from any of Examples 1-56 in Table 1.

15. A pharmaceutical composition comprising a compound according to any one of claims 1 to 14 and a pharmaceutically acceptable excipient.

16. The compound according to any one of claims 1 to 14, used in a medicament, for example for treating mitochondrial diseases.

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