EA043735B1 - PYRIDINYLSULFONAMIDE DERIVATIVES, PHARMACEUTICAL COMPOSITIONS AND THEIR APPLICATION - Google Patents

Description

Field to which the invention relates

The invention relates to new compounds, in particular pyridinylsulfonamide derivatives, to methods for producing such compounds, to their use as AOC3 inhibitors, to methods of their therapeutic use, in particular for diseases and conditions mediated by AOC3 inhibition, and to pharmaceutical compositions containing them

Prerequisites for creating the invention

The enzymatic activity of AOC3 (copper-containing amine oxidase 3; vascular adhesion protein-1) was described as early as 1967 as monoamine oxidase activity in the plasma of patients with chronic liver disease (Gressner, A. M. et al., 1982, J. Clin. Chem Clin Biochem 20: 509-514; McEwen, S. M., Jr. et al. 1967, J Lab Clin Med 70: 36-47). AOC3 has two closely homologous genes in the human genome: AOC1, which corresponds to diamine oxidase (Chassande, O. et al., 1994, J. Biol. Chem. 269: 14484-14489) and AOC2, an SSAO with specific expression in the retina (Imamura, Y. et al., 1997, Genomics 40: 277-283). AOC4 is a sequence that does not result in a functional gene product in humans due to an internal stop codon (Schwelberger, H. G., 2007, J. Neural Transm. 114: 757-762).

The enzyme contains oxidized 2,4,5-trihydroxyphenylalanine quinone (TPQ) and a copper ion in the active site. This characteristic catalytic site classifies the semicarbazide-sensitive amine oxidase (SSAO, copper-containing amine:oxygen oxidoreductase (deamination)):

Type II membrane protein belongs to the copper-containing amine oxidases family, along with several other diamines and lysyl oxidases. However, later enzymes can be distinguished from AOC3 by their preference for diamines and low sensitivity to hemicarbazide inhibition (Dunkel, P. et al., 2008, Curr. Med. Chem. 15: 1827-1839). On the other hand, monoamine oxidases contain a flavin adenine dinucleotide (FAD) cofactor in their reaction center, like monoamine oxidase A (MAO-A) and monoamine oxidase B (MAO-B), and therefore follow a different reaction pattern.

AOC3 catalyzes a two-step reaction mechanism for the oxidative deamination of primary aliphatic and aromatic amines. In the first reaction, the primary amine forms a Schiff base with the carbonyl group of TPQ. After the abstraction of a proton from the α carbon to the amino group, hydrolysis occurs and the aldehyde and aminoquinol forms of TPQ are formed in the active site. In the presence of oxygen, the aminoquinol form of TPQ is oxidized and hydrolyzed to regenerate TPQ to form ammonia and peroxide with the help of copper ion (Mure, M. et al., 2002, Biochemistry 41: 9269-9278). Several AOC3 substrates have been described, such as the physiological amines methylamine, dopamine or aminoacetone, the oxidation products of which are associated with cardiovascular pathologies (Yu, RN et al., 1993, Diabetes 42: 594-603). Synthetic amines have been optimized for turnover by AOC3, such as benzylamine derivatives (Yraola, F. et al., 2006, J. Med. Chem. 49: 6197-6208), C-naphthalene-1-methylamine (Marti, L . et al., 2004, J. Med. Chem. 47: 4865-4874) or luciferin derivatives (Valley, M. R. et al., 2006, Anal. Biochem. 359: 238-246). The latter substrate can be used for sensitive determination of AOC3 activity in plasma, tissue, or for biochemical characterization of the enzyme.

Under pathophysiological conditions of high AOC3 activity, aldehyde products are highly reactive, resulting in improved glycosylation end products (Mathys, K. S. et al., 2002, Biochem. Biophys. Res. Commun. 297: 863-869), which are considered as markers and drivers of diabetes associated with inflammatory mechanisms.

In addition, the byproduct of hydrogen peroxide is perceived by tissues as a mediator of inflammation. This reaction product is able to activate the endothelium and promotes the activation of leukocytes.

Binding and modification of Siglec-10 as a membrane-bound substrate provides mechanistic insight into how the enzymatic reaction can drive leukocyte transmigration across activated endothelium. The binding of Siglec-10 to AOC3 has been shown in several adhesion assays and resulted in increased hydrogen peroxide production (Kivi, E. et al., 2009, Blood 114: 5385-5392). Binding of activated leukocytes to dimeric extracellular AOC3 via Siglec-10 generates a transient association with activated endothelium. Consequently, the rate of leukocyte pumping is reduced, which increases the transmigration of leukocytes into the interstitium of inflamed tissues into the interstitium of inflamed tissues. In addition, the conserved RGD motif on the surface of AOC3 argues for its adhesive role: deletion of this sequence reduces leukocyte recruitment (Salmi, M. et al., 2000, Circ. Res. 86: 1245-1251), probably due to the lack of β1 integrin binding activity (Aspinall, A. I. et al., 2010, Hepatology 51: 2030-2039).

This finding correlates with the phenotype of AOC3 knockout mice, which have a reduced ability to transmigrate leukocytes and lymphocytes (Stolen, S. M. et al., 2005, Immunity. 22: 105-115) into lymphoid organs and adipose tissue (Bour, S et al., 2009, Am J Pathol 174: 1075-1083).

AOC3 activity can be detected in most tissues and is mainly expressed in endothelial cells, smooth muscle cells and adipocytes (Boomsma, F. et al., 2000, Comp Biochem. Physiol C. Toxicol. Pharmacol. 126: 69-78; O'Sullivan , J. et al., 2004, Neurotoxicology 25: 303-315).

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In humans, unlike mice, AOC3 activity is constitutive in liver sinusoidal endothelial cells (McNab, G. et al., 1996, Gastroenterology 110: 522-528) and mRNA expression is further upregulated under inflammatory conditions in this tissue (Lalor, P. F. et al., 2002, Immunol Cell Biol 80: 52-64); Bonder, C. S. et al., 2005, Immunity. 23: 153-163). AOC3 not only exists as a membrane protein, but can also be detected as soluble plasma activity, likely due to a metalloprotease-mediated shedding process (Abella, A. et al., 2004, Diabetologia 47: 429-438); Boomsma, F. et al., 2005, Diabetologia 48: 1002-1007; Stolen, S. M. et al., 2004, Circ. Res. 95: 50-57)). Increased levels of soluble AOC3 have been observed in diabetes (Li, H. Y. et al., 2009, Clin. Chim. Acta 404: 149-153), obesity (Meszaros, Z. et al., 1999, Metabolism 48: 113-117; Weiss, H. G. et al., 2003, Metabolism 52: 688-692), congestive heart failure (Boomsma, F. et al., 1997, Cardiovasc. Res. 33: 387-391), hemorrhagic stroke (HernandezGuillamon, M. et al., 2012, Cerebrovasc. Dis. 33, 55-63), end-stage renal disease (Kurkijarvi, R. et al., 2001, Eur. J. Immunol. 31: 2876-2884) and inflammatory liver disease (Kurkijarvi, R. et al. ., 1998, J. Immunol. 161: 1549-1557). For the latter, plasma AOC3 activity levels have been correlated with liver fibrosis and serve as a predictor in patients with NAFLD (Weston, S. J. et al., 2011, J. Neural Transm. 118: 1055-1064). After cirrhotic liver transplantation, high plasma AOC3 levels returned to normal values, suggesting that the liver is the main source of plasma AOC3 activity in this disease state (Boomsma, F. et al., 2003, Biochim. Biophys. Acta 1647: 48-54).

AOC3's role in inflammatory activation through peroxide production and recruitment of leukocytes to activated endothelium makes it an attractive target for treating inflammatory components in several diseases. Therefore, various small molecule compounds and antibodies have been tested in animal models of various diseases. Among them, AOC3 inhibition has demonstrated beneficial effects in cancer models of melanoma and lymphoma (Marttila-Ichihara, F. et al., 2010, J. Immunol. 184: 3164-3173), acute and chronic joint diseases (Tabi, T. et al. , 2013, J. Neural Transm. 120: 963-967) or lungs (Foot, J. S. et al., 2013, J. Pharmacol. Exp. Ther. 347: 365-374, Schilter, H. S. et al., 2015, Resp. Res. 16:42) inflammation, diabetic macular edema (Inoue, T. et al., 2013, Bioorg. Med. Chem. 21: 1219-1233), renal fibrosis (Wong, M. et al., 2014, Am. J. Physiol Renal Physiol 307: F908-F916), liver allograft rejection (Martelius, T. et al., 2004, Am. J. Pathol. 165: 1993-2001) and non-alcoholic liver disease.

Therefore, the development of a potent and well-tolerated AOC3 inhibitor will be useful for the treatment of relevant human diseases.

The enzyme copper-containing amine oxidase 2 (AOC2) is a member of a family of homodimeric amine oxidases that are sensitive to hemicarbazide inhibition. The human enzyme shares 65% of its amino acids with its closest homologue AOC3 (Zhang et al., 2003, Gene 318: 45-53). Recombinant overexpression of the longer version of sv1 provides evidence of cell surface expression and enzymatic activity, whereas the shorter version of sv2 remains cytoplasmic in the in vitro HEK293 expression system. AOC2 and AOC3 exhibit different substrate profiles due to structural differences in the active sites: AOC2 exhibits high abundance for 2-phenylethylamine and tryptamine, and low activity for methylamine or benzylamine exchange compared to the enzymatic activity of AOC3. However, both enzymes can form heterodimers that reduce enzymatic active sites while maintaining substrate selectivity. Analysis of AOC2 mRNA expression shows widespread expression of two splice variants sv1 and sv2 of the AOC2 gene in the lung, brain, heart, liver, kidney, pancreas and peripheral blood lymphocytes (Kaitaniemi et al., 2009, Cellular and Molecular Life 66: 27432757). According to AOC2 tissue enzymatic activity, the only human tissue with high AOC2-like activity is the retina, and the expression is associated with retinal capillaries as shown by immunohistochemical studies. In mice, the highest AOC2 mRNA expression is also found in the mouse retina, but the mRNA and protein expression signals are found predominantly in the retinal ganglion cell layer. In the rat, the genomic sequence of the AOC2 gene contains a stop codon in the exon 1 region, which determines the length of the peptide to be 17% of the mouse and human AOC2 protein, giving rise to a nonfunctional protein (Zhang et al., 2003, Gene 318: 45-53).

According to the enzymatic function and localization of expression, the physiological function of AOC2 may resemble the homologue AOC3, which has been described as relevant for example in neurovascular inflammation, retinal inflammation and immune cell recruitment (Matsuda et al., 2017, Invest Ophthalmol Vis Sci. 58(7) : 3254-3261, Noda et al., 2008, FASEB J. 4: 1094-103). Data on pharmacological inhibition or genetic depletion of AOC2 are not yet available, and therefore it is difficult to assess the contribution of AOC2 to retinal and vascular inflammation.

However, compared with AOC3 inhibition alone, combined inhibition of AOC2 and AOC3 may enhance anti-inflammatory effects in humans, particularly for the treatment of ocular diseases.

AOC3 inhibitors are known in the art, for example, the compounds disclosed in WO

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2013/163675, WO 2018/027892, WO 2018/148856 and WO 2018/149226. The pyridinyl sulfonamide derivatives of the present invention may provide several advantages such as increased potency, improved selectivity, reduced plasma protein binding, improved CYP enzyme profile (cytochrome P450) and high metabolic stability, high chemical stability, improved tissue distribution, e.g. effects on the brain, improved side effect profile and/or tolerability and resulting low toxicity, reduced risk of adverse events or unwanted side effects and increased solubility.

The pyridinyl sulfonamides of the present invention exhibit enhanced inhibition of human AOC2.

The pyridinyl sulfonamide derivatives of the present invention exhibit increased selectivity for AOC1. AOC1 expression and enzymatic activity are mainly found in the intestine, placenta, and kidney. The enzyme catalyzes the oxidation of primary amines obtained from food and protects humans from the cardiometabolic effects of histamine, putrescine, tryptamine and cadaverine. Inhibition of AOS1 may lead to impaired tolerance to oral histamine, resulting in increased plasma and tissue histamine levels, which may cause adverse events or undesirable side effects such as decreased blood pressure and compensation by increased heart rate, tachycardia, headache pain, hot flashes, urticaria, itching, bronchospasm and cardiac arrest (Maintz L. and Novak N. 2007. Am. J. Clin. Nutr. 85: 1185-96). The consequences of AOC1 inhibition in combination with histamine intake were demonstrated in experiments on pigs: after administration of the AOC1 inhibitor aminoguanidine (100 mg/kg) and histamine probing (2 mg/kg), an increase in the level of histamine in the blood was observed in animals, accompanied by a drop in blood pressure, increased pulse, flushing, vomiting and death (3 of 15 animals) (Sattler J. 1988. Agents and Actions, 23: 361-365) under experimental conditions. Histamine intolerance in humans has been associated with mutations in the AOC1 promoter region, resulting in decreased plasma AOC1 mRNA expression and activity (Maintz et al. 2011. Allergy 66: 893-902).

Objective of the present invention

The object of the present invention is to provide new compounds, in particular new pyridinyl sulfonamide derivatives, which are active against AOC2 and AOC3.

Another object of the present invention is to provide new compounds, in particular new pyridinyl sulfonamide derivatives, which have an inhibitory effect on AOC2 and AOC3 in vitro and/or in vivo, and have suitable pharmacological and pharmacokinetic properties for use as drugs.

Another object of the present invention is to provide effective dual inhibitors of AOC2 and AOC3, in particular for the treatment of various diseases, for example cancer, NASH (non-alcoholic steatohepatitis), pulmonary fibrosis, retinopathy, nephropathy and stroke, in particular hemorrhagic stroke.

Another object of the present invention is to provide effective dual inhibitors of AOC2 and AOC3 for the treatment of metabolic disorders such as cancer, NASH (non-alcoholic steatohepatitis), pulmonary fibrosis, retinopathy, nephropathy and stroke, in particular hemorrhagic stroke.

Another object of the present invention is to provide methods for treating a disease or condition mediated by inhibition of AOC2 and AOC3 in a patient.

Another object of the present invention is to provide a pharmaceutical composition containing at least one compound according to the invention.

Another object of the present invention is to provide a combination of at least one compound according to the invention with one or more additional therapeutic agents.

Another object of the present invention is to provide methods for the synthesis of new compounds, in particular pyridinyl sulfonamide derivatives.

Another object of the present invention is to provide starting materials and/or intermediates useful in methods for the synthesis of new compounds.

Other objects of the present invention will become apparent to one skilled in the art from the description above and below, as well as from the examples.

Object of the invention

Within the framework of the present invention, it has surprisingly been discovered that the new compounds of general formula (I) described below exhibit inhibitory activity against AOC2 and AOC3.

According to another aspect of the present invention, the novel compounds of general formula (I) described below have been found to exhibit AOC3 inhibitory activity.

In a first aspect, the present invention relates to a compound of the general formula

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in which ring A is selected from the group A-G1 consisting of:

* [^^N—* ^< ^3 ^N ''* ^< \ / ^N ~ * , , and *

R1 is selected from the group R1-G1, consisting of H, F, Cl, Br, CN, -OH, C _1-4 -alkyl, -O-(C _1-4 -alkyl), -(CH2) _t -COOH, -(CH2) _t -C(=O)-O-(C1-4-alkyl), -C(=O)-heterocyclyl, -(CH2) _m -C(=O)-NH2, -(CH2)m -C(=O)NH-(C _1-4 -alkyl), -(CH ₂ ) _m -C(=O)-N(C _1-4 -alkyl) ₂ , -C(=O)-NH- C _3-6 -cycloalkyl, -C(=O)-NH-heterocyclyl, -(CH ₂ ) _m -NH-C(=O)-(C ₁ .3-alkyl), -N(C _1-3 - alkyl)-C(=O)-(C _1-4 -alkyl), -N(C _1-3 -alkyl)-C(=O)-NH ₂ , -NHC(=O)-NH-(C _{1 -4} -alkyl), heterocyclyl and phenyl, wherein each alkyl group or subgroup is optionally substituted by 1 or more F atoms or one OH or -O-(C _1-3 -alkyl) group; and wherein each heterocyclyl is selected from the group consisting of azetidinyl, imidazolidinyl, piperidinyl, tetrahydropyranyl and morpholinyl and is optionally substituted with one or two groups independently selected from the group consisting of oxo, C _1-3 -alkyl, -C(=O)- CH ₃ and -C(=O)-cyclopropyl; and where several R ¹ can be the same or different if n is 2; and n is an integer selected from 1 and 2; and m is an integer selected from 0, 1 and 2; and wherein in any definition mentioned above, unless otherwise stated, any alkyl group or subgroup may be linear or branched and is optionally substituted by 1 or more F atoms, a tautomer or stereoisomers thereof, or a salt thereof, or a solvate or hydrate thereof.

In a further aspect, the present invention relates to methods for preparing a compound of general formula (I) and to new intermediates in these methods.

Another aspect of the invention relates to a salt of the compounds of general formula (I) according to the present invention, in particular to a pharmaceutically acceptable salt thereof.

In another aspect, the present invention relates to a pharmaceutical composition containing one or more compounds of general formula (I) or one or more pharmaceutically acceptable salts thereof according to the invention, optionally together with one or more inert carriers and/or diluents.

In another aspect, the present invention relates to a method of treating diseases or conditions that are mediated by inhibition of AOC3 activity in a patient in need thereof, characterized in that a compound of general formula (I) or a pharmaceutically acceptable salt thereof is administered to the patient.

According to another aspect of the invention, there is provided a method of treating cancer, NASH (non-alcoholic steatohepatitis), pulmonary fibrosis, retinopathy, nephropathy or stroke in a patient in need thereof, characterized in that a compound of general formula (I) or a pharmaceutically acceptable salt thereof is administered to the patient.

According to another aspect of the invention, there is provided the use of a compound of general formula (I) or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for a therapeutic method as described above or below.

According to another aspect of the invention, there is provided a compound of general formula (I) or a pharmaceutically acceptable salt thereof for use in a therapeutic method as described above or below.

In a further aspect, the present invention relates to a method of treating a disease or condition mediated by AOC3 inhibition in a patient, which includes the step of administering to a patient in need of such treatment a therapeutically effective amount of a compound of general formula (I) or a pharmaceutically acceptable salt thereof in combination with a therapeutically effective amount of one or several additional therapeutic agents.

In another aspect, the present invention relates to the use of a compound of general formula (I) or a pharmaceutically acceptable salt thereof in combination with one or more additional therapeutic agents for the treatment or prevention of diseases or conditions that are mediated by AOC3 inhibition.

In another aspect, the present invention relates to a pharmaceutical composition that contains a compound according to general formula (I) or a pharmaceutically acceptable salt thereof and one or

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Other aspects of the invention will become apparent to one skilled in the art from the description and experimental portions as described above and below in the present invention.

Detailed description

Unless otherwise indicated, groups, residues and substituents, in particular A, R ¹ and R ² , have the definitions given above and below. If residues, substituents or groups occur several times in a compound, such as R ² , they may have the same or different meanings. Some preferred meanings of individual groups and substituents of the compounds according to the invention will be given below. Any and all of these definitions can be combined with each other.

A.

A-G1.

Ring A is preferably selected from the group A-G1 as defined above.

A-G2.

In another embodiment, ring A is selected from the group A-G2 consisting of oA-G3.

In another embodiment, ring A is selected from the group A-G3 consisting of *

A-G4.

In another embodiment, ring A is selected from the group A-G4 consisting of <Q ^N -'

A-G5.

In another embodiment, ring A is selected from the group A-G5 consisting of <Qn-*

^R1 .

R ¹ -G1.

The R ¹ group is preferably selected from the R1-G1 group as defined above.

R ¹ -G1a.

In one embodiment, the group R ¹ is selected from the group R1-G1a, including:

H, F, Cl, -OH, C _1-4 -alkyl, -O-(C _1-2 -alkyl), -(CH ₂ ) _m -COOH, -(CH ₂ ) _m -C(=O)- O-(C ₁ . ₂ -αlkyl), -C(=O)heterocyclyl, -(CH ₂ ) _m -C(=O)-NH ₂ , -(CH ₂ ) _m -C(=O)-NH- (C _1-4 -alkyl), -(CH ₂ ) _m -C(=O)-N(CH ₃ )(C _1-3 -alkyl), -C(=O)-NH-cyclopropyl, -C( =O)-NH-heterocyclyl, -(CH ₂ ) _m -NH-C(=O)-(C _1-3 -alkyl), -N(C _1-2 -alkyl)C(=O)-(C _1-2 -alkyl), -N(C _1-2 -alkyl)-C(=O)-NH ₂ , -NH-C(=O)-NH-(C _1-2 -alkyl), heterocyclyl and phenyl wherein each alkyl group or subgroup is optionally substituted with 1 to 3 F atoms or one OH or -O-(C _1-2 -alkyl) group; and wherein each heterocyclyl is selected from the group consisting of azetidinyl, imidazolidinyl, piperidinyl, tetrahydropyranyl and morpholinyl and is optionally substituted with one or two groups independently selected from the group consisting of oxo, C _1-2 -alkyl, -C(=O)- CH ₃ and -C(=O)-cyclopropyl; and where m means 0 or 1; and where several R ¹ can be the same or different if n is 2.

R ¹ -G1b.

In another embodiment, the group R ¹ is selected from the group R1-G1b, including:

H, F, -OH, -CH3, -CF ₃ , -O-CH3, -COOH, -(CH2)m-C(=O)-O-CH3, -(CH2)mC(=O)-NH2, -C(=O)-NH-(Ci, ₃ alkyl), -(CH2)-C(=O)-N(CH3)2, -(CH2)-C(=O)^(CH3)(CH2CH3) , -C(=O)-NH-cyclopronyl, 1(cyclopropylcarbonyl)-piperidin-4-yl and 3-methyl-2-oxo-imidazolidin-1-yl, wherein each ethyl group or subgroup is optionally substituted at the 2-position by one atom F, one OH or one -O-CH3 group; and wherein each propyl group or subgroup is optionally substituted at the 2 or 3 position with 1 to 3 F atoms; and where m means 0 or 1; and where several R ¹ can be the same or different if n is 2.

If n is 2, then the second R ¹ group of R1-G1, R1-G1a or R1G1b is preferably selected from the group consisting of F, CH ₃ , CF ₃ and phenyl.

In another embodiment, the group R ¹ is selected from the group R1-G2, including:

H, F, -OH, C _1-4 -alkyl, -O-(C _1-4 -alkyl), -(CH ₂ ) _m -COOH, -(CH ₂ ) _m -C(=O)-O- (C _1-4 -alkyl), -C(=O)heterocyclyl, -(CH ₂ ) _m -C(=O)-NH ₂ , -(CH ₂ ) _m -C(=O)-NH-(Cl _-4 -alkyl), -(CH ₂ ) _m -C(=O)-N(C _1-4 -alkyl) ₂ , -C(=O)-

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NH-C _3-6 -cycloalkyl, -C(=O)-NH-heterocyclyl, -(CH2) _m -NH-C(=O)-(C ₁ .3-αlkul), -M(C _1-3 -alkyl)-C(=O)-(C ₁ . ₄ alkyl), -N(C _1-3 -alkyl)-C(=O)-NH ₂ , -NH-C(=O)-NH-( C _1-4 -alkyl), heterocyclyl and phenyl, wherein each alkyl group or subgroup is optionally substituted by 1 or more F atoms or one OH or -O-(C _1-3 -alkyl) group; and wherein each heterocyclyl is selected from the group consisting of azetidinyl, imidazolidinyl, piperidinyl, tetrahydropyranyl and morpholinyl and is optionally substituted with one or two groups independently selected from the group consisting of oxo, C _1-3 -alkyl, -C(=O)- CH ₃ and -C(=O)-cyclopropyl; and where several R ¹ can be the same or different if n is 2.

R ¹ -G2a.

In another embodiment, the group R ¹ is selected from the group R1-G2a, including:

H, -OH, C _1-2 -alkyl, -O-(C _1-2 -alkyl), -(CH ₂ ) _m -COOH, -(CH ₂ ) _m -C(=O)-O-(C _1-2 -alkyl), -C(=O)heterocyclyl, -(CH ₂ ) _m -C(=O)-NH ₂ , -(CH ₂ ) _m -C(=o)-NH-(C _{1- 4} -alkyl), -(CH ₂ ) _m -C(=O)-N(C _1-2 -alkyl) ₂ , -C(=O)NH-C _3-6 -cyclopropyl, -C(=O) -NH-heterocyclyl, -(CH ₂ ) _m -NH-C(=O)-(C _1-3 alkyl), -N(CH ₃ )-C(=O)-(c _1-2 alkyl), -N(CH ₃ )-C(=O)-NH ₂ , -NH-C(=O)-NH-(C _1-3 -alkyl), heterocyclyl and phenyl, where each alkyl group or subgroup is optionally substituted by 1 up to 3 F atoms or one OH or -O-CH ₃ group; and wherein each heterocyclyl is selected from the group consisting of azetidinyl, imidazolidinyl, tetrahydropyranyl and morpholinyl and is optionally substituted with one or two groups independently selected from the group consisting of oxo, C _1-3 -alkyl and -C(=O)-CH ₃ ; and where several R ¹ can be the same or different if n is 2.

R1-G2b.

In another embodiment, the group R ¹ is selected from the group R1-G2b consisting of:

H, -OH, C1-2-alkyl, -O-CH ₃ , -(CH^-COOH, -(CH2)m-C(=O)-O-CH3, -C(=O)-heterocyclyl, - (CH2)mC(=O)-NH2, -(CH2)m-C(=O)-NH-(C1-4-alkyl), -(CH2)mC(=O)-N(CH3)2, - C(=O)-NH-C3.6-cyclonronyl, -C(=O)-NH-τtetrahydropyranyl, -(CH ₂ ) _m -NH-C(=o)-(C _1-2 -alkyl), - N(CH ₃ )-C(=O)-CH ₃ , -N(CH ₃ )-C(=O)-NH2, -NH-C(=O)-NH-CH ₃ , imidazolidinyl and phenyl, where each the alkyl group or subgroup is optionally substituted with 1 to 3 F atoms or one OH group; and wherein the imidazolidinyl group is optionally substituted with one or two groups independently selected from the group consisting of oxo and CH ₃ , and wherein each heterocyclyl is selected from the group consisting of itself azetidinyl and morpholinyl and optionally substituted with one CH3; and where m is 0 or 1; and where several R ¹ may be the same or different if n is 2.

The groups R1-G2, R1-G2a and R1-G2b are preferably combined with the group A-G2.

if n is 2, then the second R ¹ group of R1-G2, R1-G2a or R1-G2b is preferably selected from the group consisting of CH3, CF3 and phenyl.

R ¹ -G3.

In another embodiment, the group R ¹ is selected from the group R1-G3, including:

H, F, Cl, -OH, -O-(C _1-4 -alkyl), -C(=O)-heterocyclyl, -(CH ₂ ) _m -C(=O)-NH ₂ , -(CH ₂ ) _m -С(=О)-NH-(С _1-4 alkyl), -(СН ₂ ) _m -С(=О)-N(С _1-4 -alkyl) ₂ , -(СН ₂ ) _m - NH-C(=O)-(C _1-3 -alkyl) and -N(c _1-3 -alkyl)-C(=O)-(C _1-4 -alkyl), where each alkyl group or subgroup is optional substituted with 1 or more F atoms or one OH or -O-(C _1-3 -alkyl) group; and wherein each heterocyclyl is selected from the group consisting of azetidinyl, imidazolidinyl, piperidinyl, tetrahydropyranyl and morpholinyl and is optionally substituted with one oxo or C _1-3 alkyl group; and where several R ¹ can be the same or different if n is 2.

R ¹ -G3a.

In another embodiment, the group R ¹ is selected from the group R1-G3a, including:

H, F, -OH, -O-(C _1-2 -alkyl), -C(=O)-morpholinyl, -C(=O)-NH ₂ , -C(=O)-NH-(C _{1 -4} -alkyl), -C(=O)-N(C _1-3 alkyl) ₂ , -NH-C(=O)-(C _1-2 -alkyl) and -N(CН ₃ )-C( =O)-(C ₁ - ₂ -alkyl), where each alkyl group or subgroup is optionally substituted with 1 to 3 F atoms or one OH or -O-(C _1-3 -alkyl) group; and where several R ¹ can be the same or different if n is 2.

R ¹ -G3b.

In another embodiment, the group R ¹ is selected from the group R1-G3b, including:

H, F, -OH, -O-CH3, -C(=O)-morpholinyl, -C(=O)-NH2, -C(=O)-NH-(C1-4-alkyl), -C( =O)-I(CH3) ₂ and -NHC(=O)-(CH3), where each alkyl group or subgroup is optionally replaced by one OH group; and where several R ¹ can be the same or different if n is 2.

The groups R1-G3, R1-G3a and R1-G3b are preferably combined with the group A-G3.

if n is 2, then the second group R ¹ of R1-G3, R1-G3a or R1-G3b is preferably F.

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In another embodiment, the group R ¹ is selected from the group R1-G4, including:

H, -(CH ₂ ) _m -COOH, -(CH ₂ ) _m -C(=O)-O-(C ₁ . ₄ -alkyl), -C(=O)-heterocyclyl, -(CH ₂ ) _m -C(=O)-NH ₂ ,

-(CH ₂ ) _m -C(=O)-Nн-(C ₁ . ₄ -alkyl) and -(cH ₂ ) _m -C(=O)-N(C _1-4 -alkyl) ₂ , where each the alkyl group or _subgroup is optionally substituted with 1 or more F atoms or one OH or -O-(C _{1_3} -alkyl) group; and wherein each heterocyclyl is selected from the group consisting of azetidinyl, imidazolidinyl _, piperidinyl, tetrahydropyranyl and morpholinyl and is optionally substituted with one oxo or C _{1_3} -alkyl group; and where several R ¹ can be the same or different if n is 2.

R ¹ -G4a.

In another embodiment, the group R ¹ is selected from the group R1-G4a, including:

H, -COOH, -C(=O)-O-( _C1.2 -alkyl), -C(=O) _-morpholinyl , -C(=O) _-NH2 , -C(=O)-NH -(C ₁ . ₄ -alkyl) and -C(=O)-N(C ₁ . ₄ -alkyl) ₂ , where each alkyl group or subgroup is optionally substituted with 1 to 3 F atoms or one OH or -O-( With _1-3 -alkyl) group; and where several R ¹ can be the same or different if n is 2.

R ¹ -G4b.

In another embodiment, the group R ¹ is selected from the group R1-G4b, including:

H, -COOH, -C(=O)-O-CH ₃ , -C(=O)-morpholinyl, -C(=O)-NH2, -C(=O)-NH-(C _1-4 - αlkyl) and -C(=O)-N(CH ₃ )(C _1-4 alkyl), where each alkyl group or subgroup is optionally replaced by one -O-CH ₃ group. The groups R1-G4, R1-G4a and R1-G4b are preferably combined with the group A-G4.

If A is selected from A-G4, then n is preferably 1.

R ¹ -G5.

In one embodiment, the group R ¹ is selected from the group R1-G5, consisting of:

H, F, Cl, Br, CN, -OH, C ₁ . ₄ -alkyl, -O-(C ₁ . ₄ -alkyl), -C(=O)-NH ₂ , -C(=O)-NH-(C ₁ . ₄ -alkyl), -C^JSalkyl) ₂ and heterocyclyl, wherein each alkyl group or _subgroup is optionally substituted with 1 or more F atoms or one OH or -O-(C _{1_3} -alkyl) group; and wherein each heterocyclyl is selected from the group consisting of azetidinyl and piperidinyl, and is optionally substituted with one C1_ ₃ -alkyl, -C(=O)-CH ₃ or -C(=O)-cyclopropyl group; and where several R ¹ can be the same or different if n is 2.

R ¹ -G5a.

In another embodiment, the group R ¹ is selected from the group R1-G5a, including:

H, F, -OH, C ₁ . ₄ -alkyl, -O-(C ₁ - ₂ -alkyl), -C(=O)-NH ₂ , -C(=O)-NH-(C ₁ . ₂ -alkyl), -C(=O) -N(C ₁ . ₂ -alkyl) ₂ and piperidinyl, wherein each alkyl group or subgroup is optionally substituted with 1 to 3 F atoms or one OH group; and wherein the piperidinyl group is optionally substituted with one -C(=O)-CH ₃ or -C(=O)cyclopropyl group; and where several R ¹ can be the same or different if n is 2.

R ¹ -G5b.

In another embodiment, the group R ¹ is selected from the group R1-G5b, including:

H, F, -OH, C ₁ . _4- alkyl, -O-CH ₃ , -C(=O)-NH ₂ , -C(=O)-NH-(CH ₃ ), -C(=O)-N(CH ₃ )2 and piperidinyl, wherein each alkyl group or subgroup is optionally substituted with one OH group; and wherein the piperidinyl group is optionally substituted with one -C(=O)-cyclopropyl group; and where several R ¹ can be the same or different if n is 2.

The groups R1-G5, R1-G5a and R1-G5b are preferably combined with the group A-G5.

if n is 2, then the second group R ¹ of R1-G5, R1-G5a or R1-G5b is preferably selected from the group consisting of F and CH ₃ .

n.

In one embodiment, n is an integer selected from 1 and 2. Preferably, n is 1.

In another embodiment, n is 2.

m.

In one embodiment, m is an integer selected from 0, 1, and 2. Preferably, m is 0 or 1.

In another embodiment, m is 0.

In yet another embodiment, m is 1.

The following preferred embodiments of the compounds of formula I are described using the general formulas I.1-I.4, which include any tautomers, solvates, hydrates and salts thereof, in particular pharmaceutically acceptable salts thereof.

- 7 043735

In the above formulas (1.1)-(1.4), n and the group R ¹ have the above definitions.

Examples of preferred sub-generic embodiments (E) in accordance with the present invention are set forth in the following table, in which each group of substituents for each embodiment is defined in accordance with the definitions set forth above:

Embodiment Formula A R ¹ n E1 I A-G1 R^Gl 1 or 2 E2 I A-G1 R<G1 1 EZ I A-G1 R^Gla 1 or 2 E4 I A-G1 R<Gla 1 E5 I A-G1 R^Glb 1 or 2 E6 I A-G1 R ⁱ -Glb 1 E7 I A-G2 R<G2 1 or 2 E8 I A-G2 R<G2 1 E9 I A-G2 R<G2a 1 or 2 EY I A-G2 R<G2a 1 E11 I A-G2 R ^£ -G2b 1 or 2 E12 I A-G2 R<G2b 1 E13 I A-G3 R<G3 1 or 2 E14 I A-G3 R<G3 1 E15 I A-G3 R<G3a 1 or 2 E16 I A-G3 R ⁱ -G3a 1 E17 I A-G3 R ^L -G3b 1 or 2 E18 I A-G3 R ⁱ -G3b 1 E19 I A-G4 R<G4 1 or 2 E20 I A-G4 R<G4 1 E21 I A-G4 R ⁱ -G4a 1 or 2 E22 I A-G4 R<G4a 1 E23 I A-G4 R ^L -G4b 1 E24 I A-G5 R<G5 1 or 2 E25 I A-G5 R<G5 1 E26 I A-G5 R<G5a 1 or 2 E27 I A-G5 R<G5a 1 E28 I A-G5 R ^L -G5b 1 or 2 E29 I A-G5 R<G5b 1

Preferred compounds according to the invention include:

- 8 043735

- 9 043735

- 10 043735 and salts thereof, preferably pharmaceutically acceptable salts thereof.

Particularly preferred compounds, including tautomers and stereoisomers thereof, salts thereof, or any solvates or hydrates thereof, are described in the experimental section below.

The compounds of the invention can be prepared using synthetic methods known to one skilled in the art and described in the organic synthesis literature. Preferably, the compounds are prepared analogously to the preparation methods described in more detail below, in particular as described in the experimental section.

Concepts and definitions.

Terms not specifically defined in the present invention are to be given the meanings given to them by one skilled in the art in light of the disclosure and context. However, as used in the specification, unless otherwise specified, the following terms have the meaning indicated and the following conventions apply.

The terms compound(s) of the present invention, compound(s) of formula (I), compound(s) of the invention, and the like refer to the compounds of formula (I) provided herein, including tautomers, stereoisomers, and mixtures thereof, and their salts, in particular their pharmaceutically acceptable salts, and solvates and hydrates of such compounds, including solvates and hydrates of such tautomers, stereoisomers and salts thereof.

Notwithstanding the above, the compounds of the invention always have the E configuration in the vinyl fluoride moiety.

The terms treatment and treat cover both preventative, i.e. prophylactic, and therapeutic, i.e. curative and/or palliative treatment. Thus, the terms treatment and treat refer to the therapeutic treatment of patients who have already developed the specified condition, in particular in an overt form. Therapeutic treatment may be symptomatic treatment to relieve symptoms of a particular indication or causative treatment to completely or partially change the conditions of the indication or to stop or slow the progression of a disease. Thus, the compositions and methods in accordance with the present invention can be used, for example, as a therapeutic treatment over a period of time, as well as for long-term therapy. In addition, the terms treat and treat include preventive treatment, that is, treatment of patients at risk of developing a condition mentioned above, thereby reducing said risk.

If the present invention relates to patients in need of treatment, it primarily relates to the treatment of mammals, in particular humans.

The term "therapeutically effective amount" means an amount of a compound of the present invention that (i) treats or prevents a particular disease or condition, (ii) mitigates, ameliorates, or eliminates one or more symptoms of a particular disease or condition, or (iii) prevents or delays the onset of one or more than one symptom of a particular disease or condition described in the present invention.

As used herein, the terms modulated or modulating, or modulate, unless otherwise specified, refer to the inhibition of AOC3 by one or more compounds of the present invention.

- 11 043735

As used herein, the terms indirect or mediate or mediate, unless otherwise specified, refer to (i) treatment, including prevention of a particular disease or condition, (ii) alleviation, improvement or elimination of one or more symptoms of a particular disease or condition, or ( iii) preventing or delaying the onset of one or more symptoms of a particular disease or condition described in the present invention.

As used herein, the term substituted means that any one or more hydrogen atoms on a specified atom, radical or moiety are replaced by those selected from a specified group, provided that the normal valency of the atom is not exceeded and that the substitution results in an acceptably stable compound.

In the groups, radicals or moieties defined below, the number of carbon atoms is often indicated before the group, for example, C _1-6 -alkyl means an alkyl group or radical having from 1 to 6 carbon atoms. Typically, for groups containing two or more subgroups, the last named subgroup is the point of addition of the radical, for example, the substituent aryl-C _1-3 -alkyl- means an aryl group that is bonded to the C _1-3 -alkyl group, the last of which associated with the nucleus or with a group to which a substituent is attached.

Where a compound of the present invention is depicted in both chemical name and formula form, in case of any discrepancy, the formula shall prevail.

An asterisk can be used in subformulas to indicate a bond that is connected to the main molecule as defined.

The numbering of substituent atoms begins with the atom that is closest to the nucleus or group to which the substituent is attached.

For example, the term 3-carboxypropyl group represents the following substituent:

where the carboxy group is attached to the third carbon atom of the propyl group. The terms 1methylpropyl, 2,2-dimethylpropyl or cyclopropylmethyl group represent the following groups:

An asterisk can be used in subformulas to indicate a bond that is connected to the main molecule as defined.

In the definition of a group, the expression where each group X, Y and Z is optionally substituted by and the like means that each group X, each group Y and each group Z, either each as a separate group, or each as part of a composite group, can be substituted , as defined. For example, the definition of R ^ex means H, C1-3-alkyl, C3-6-cycloalkyl, C3-6-cycloalkyl-C1-3-alkyl, or C1-3-alkyl-O-, each alkyl group being optionally substituted by one or more L ^ex or the like means that in each of the above groups which include the term alkyl, i.e. in each of the C1-3-alkyl, _C3-6 -cycloalkyl- _C1-3 -alkyl and _C1-3 -alkyl-O- groups, the alkyl moiety may be substituted by L ^ex as defined.

In the following, the term bicyclic includes spirocyclic.

Unless specifically stated, throughout the specification and appended claims, the chemical formula or name is intended to cover tautomers and all stereo, optical and geometric isomers (e.g., enantiomers, diastereomers, etc.) and racemates thereof, as well as mixtures in various ratios of individual enantiomers, mixtures of diastereomers or mixtures of any of the above forms, where such isomers and enantiomers exist, as well as salts, including pharmaceutically acceptable salts thereof and solvates thereof, such as, for example, hydrates, including solvates of the free compounds or solvates of a salt of a compound. Notwithstanding the above, the compounds of the invention always have the E configuration in the vinyl fluoride moiety.

As used herein, the expression pharmaceutically acceptable refers to those compounds, substances, compositions and/or dosage forms that are medically suitable for use in contact with tissues of humans and animals without undue toxicity, irritation, allergic response or other difficulty, or complications, and are commensurate with a reasonable benefit/risk ratio.

As used herein, the term pharmaceutically acceptable salts refers to derivatives of the disclosed compounds in which the parent compound is modified to form acidic or basic salts thereof. Examples of pharmaceutically acceptable salts include, but are not limited to, salts of mineral or organic acids with basic moieties such as

- 12 043735 amines; alkaline or organic salts of acid residues such as carboxylic acids; etc.

The pharmaceutically acceptable salts of the present invention can be synthesized from the starting compound which contains the acid moiety by conventional chemical methods. Typically, such salts can be prepared by reacting the free acid forms of these compounds with a sufficient amount of the corresponding base in water or an organic diluent such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile, or a mixture thereof.

Salts of other acids other than those mentioned above, which are, for example, useful for purifying or isolating compounds according to the present invention (eg trifluoroacetate salts) are also part of the invention.

The term halogen usually means fluorine, chlorine, bromine and iodine.

Term C ₁ . _n -alkyl, where n is an integer from 1 to n, alone or in combination with another radical, means an acyclic, saturated, branched or linear hydrocarbon radical with from 1 to n C atoms. For example, the term C ₁ . ₅ -alkyl covers the radicals H ₃ C-, H3C-CH2-, H ₃ CH2-CH2-, H3C-CH(CH3)-, H3C-CH2-CH2-CH2-, H3C-CH2-CH(CH3)-, H3C -CH(CH3)-CH2-, H3C-C( _CH3 )2-, H3C-CH2-CH2-CH2-CH2-, H3C-CH2-CH2-CHCH3)-, H3C-CH2-CH(CH3)-CH2 -, НзС-СН(СНз)-СН2-СН2-, Н3СН2-С(СНз)2-, НзС-С(СНз)2-СН2-, НзС-СН(СНз)-СН(СНз)- and НзС-СН ₂ -CH(CH ₂ CH3)-.

Term C ₃ . _n -cycloalkyl, where n is an integer from 4 to n, either alone or in combination with another radical, means a cyclic, saturated, unbranched hydrocarbon radical with from 3 to n C atoms. The cyclic group may be mono-, bi-, tri- or spirocyclic, most preferably monocyclic. Examples of such cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclododecyl, bicyclo[3.2.1.]octyl, spiro[4.5]decyl, norpinyl, norbonyl, norcaryl, adamantyl, etc.

Many of the terms above may be used multiple times in the definition of a formula or group, and in each case have one of the meanings given above, independently of each other.

All residues and substituents, as defined above and below, may be replaced by one or more F atoms.

Pharmacological activity.

The activity of the compounds can be demonstrated using the following AOC3 assay. Biochemical analysis of AOS3.

The MAO-Glo™ assay (commercially available from PROMEGA, #V1402) provides a sensitive method for measuring monoamine oxidase (MAO) activity (Valley, M.R. et al., 2006, Anal. Biochem. 359: 238-246) from a variety of tissues, biological fluids or recombinant expressed or purified enzymes. The substrate used is a beetle luciferin derivative ((4S)-4,5dihydro-2-(6-hydroxybenzothiazolyl)-4-thiazole-carboxylic acid), which is oxidized at the primary amine fragment. Following spontaneous removal and catalyzed esterase reaction, luciferin turnover by luciferase is detected as a signal of AOC3 activity.

To determine AOC3 activity or inhibitory potency of a compound, inhibitory compounds are dissolved in DMSO and adjusted to the appropriate assay concentration using reaction buffer (50 mM HEPES, 5 mM KCl, 2 mM CaCl ₂ , 1.4 mM MgCl ₂ , 120 mM NaCl, 0.001% (v/v) Tween 20, 100 µM TCER, pH 7.4). A 3 µl aliquot of the compound dilution was added to a 384 well plate (Optiplate, PS, flat bottom, white, PERKIN ELMER, #6007290) with a final DMSO concentration of 6.6%. Recombinant CHO cells overexpressing human (1500 cells/well), mouse (1000 cells/well) or rat (500 cells/well) AOC3 enzyme are diluted in the reaction buffer and added to the wells in a volume of 15 μl. After incubation for 20 min at 37°C, add 2 μl of MAO substrate (dissolved in DMSO at 16 mM, adjusted to the analytical concentration in the reaction buffer to a final analytical concentration of 20 μM) and further incubate for 60 min at 37°C. Substrate turnover was determined by adding 20 μl of detection mixture, which was prepared by adding esterase reduction buffer (PROMEGA, #V1402) to luciferin detection reagent (PROMEGA, #V1402). After a 20-minute incubation period, the luminescence signal is measured using an Envision 2104 Multilabel Reader (PERKIN ELMER).

Alternative assays to determine AOC3 enzymatic activity include extraction with ¹⁴ C-labeled benzylamine or the Amplex Red monoamine oxidase reaction (Molecular Probes, The Netherlands) as described by Gella et al. (Gella, A. et al., 2013, J. Neural Transm. 120: 1015-1018).

The compounds of general formula (I) according to the invention, for example, have IC ₅₀ values below 5000, especially below 1000, preferably below 300, most preferably below 100 nM.

Biochemical analysis of AOS2.

Amplex® Red Assay (commercially available from Thermo Fisher Scientific) provides a sensitive method for detecting H2O2 produced during enzymatic reactions such as AOC2-catalyzed amine oxidation. The assay reagent is a colorless substrate (Nacetyl-3,7- _{dihydroxyphenoxazine} ), which reacts in a 1:1 stoichiometry with hydrogen peroxide ( _H2O2 ) to form the fluorescent dye resorufin (7-hydroxyphenoxazin-3-one, excitation maxima/ emission = 570/585 nm).

To determine AOC2 activity or the inhibition efficiency of an AOC2 compound, inhibitory compounds are dissolved in DMSO and adjusted to the appropriate 20-fold concentration using reaction buffer (100 mM sodium phosphate, 0.05% Pluronic F-127 (#P3000MP Sigma-Aldrich, pH 7 ,4) A 5 µL aliquot of the compound dilution is added to a 96-well plate (flat bottom F, black, GREINER bio-one, #655900) at a DMSO concentration of 2%.

AOC2 enzyme containing cell homogenate is obtained by transiently transfecting 6x106 HEK293 cells per vial (T75) with 9 μg pCMV-SPORT6-AOC2 (BC142641rc, #pCS6(BC142641)-seqTCHS1003-GVO-TRI, BioCat) in 750 μl EMEM culture medium (#BE12-611F, Lonza) and 33.75 µl Attractene (#301005, Qiagen). Cells were cultured for 3 days in EMEM culture medium containing 10% TPS (#04-00-1A, Biological Industries). After washing twice with ice-cold PBS, the cells were lysed by mechanical homogenization and the purified supernatants were shock frozen in liquid nitrogen and stored at -80°C.

To determine the enzymatic activity of AOC2, cell lysates are thawed on ice and diluted 1:1 with reaction buffer. A 45 µl aliquot is added to the diluted compound and incubated for 30 min at 37°C. The enzymatic reaction begins with the addition of 50 µl of Amplex® Red reaction mixture (final assay concentration: 100 mM sodium phosphate, 120 µM Amplex® Red reagent (#A22177 Molecular Probes), 1.5 U/ml horseradish peroxidase (#P8375 Sigma-Aldrich) , 2 mM phenylethylamine (#P6513-25G Sigma-Aldrich), 0.05% Pluronic F-127 (#P3000MP Sigma-Aldrich), pH 7.4, 37°C).

Substrate turnover over time is determined directly using a fluorescent reader (Ex 540 nm/Em 590 nm) such as an Envision 2104 Multilabel Reader (PERKIN ELMER) for 60 min. (See Anal Biochem (1997) 253:169-174; Anal Biochem (1997) 253:162-168).

Biochemical analysis of AOS1.

Amplex® Red Assay (available from Thermo Fisher Scientific) is a sensitive method for detecting H2O2 produced during enzymatic reactions such as AOC1-catalyzed amine oxidation. The assay reagent is a colorless substrate (M-acetyl- _{3,7dihydroxyphenoxazine} ), which reacts in a 1:1 stoichiometry with hydrogen peroxide ( _H2O2 ) to form the fluorescent dye resorufin (7-hydroxyphenoxazin-3-one, excitation maxima/ emission = 570/585 nm).

To determine AOC1 activity or AOC1 inhibitory potency of a compound, inhibitory compounds are dissolved in DMSO and adjusted to the appropriate concentration using reaction buffer (100 mM sodium phosphate, 0.05% Pluronic F-127 (#P3000MP Sigma-Aldrich), pH 7.4) . A 3 μl aliquot of the compound dilution is added to a 384-well plate (Optiplate, PS, flat bottom F, black, PERKIN ELMER, #6007270) at a DMSO concentration of 6.6%.

An aliquot of AOC1 enzyme (#8297-AO-010, R&D Systems) was thawed on ice, diluted in reaction buffer, and added in a volume of 7 μL to the wells to obtain a final assay concentration of 1 ng/well. After incubating the inhibitor and enzyme for 30 min at 37°C, the enzymatic reaction begins with the addition of 10 µl of Amplex® Red reaction mixture (final assay concentration: 100 mM sodium phosphate, 120 µM Amplex® Red reagent (#A22177 Molecular Probes), 1, 5 U/ml horseradish peroxidase (#P8375 Sigma-Aldrich), 200 µM putrescine (#P7505 Sigma-Alrdich), 0.05% Pluronic F127 (#P3000MP Sigma-Aldrich), pH 7.4, 37°C).

After incubation for 30 minutes at 37°C, substrate turnover is determined directly (or after adding excess amine oxidase inhibitor) using a fluorescence reader (Ex 540 nm/Em 590 nm) such as an Envision 2104 Multilabel Reader (PERKIN ELMER).

The table below presents the activity expressed as IC ₅₀ (nM) of the compounds according to the invention, where the IC ₅₀ values are determined in the AOC3, AOC2 and AOC1 assays as described above. The term Example refers to the example numbers according to the following experimental section.

Biological data of the compounds of the present invention obtained in the AOC3, AOC2 and AOC1 assays. But. = not defined.

- 14 043735

Example AOSZ 1C ₅₀ AOS2 1C ₅₀ AOS1 1C ₅₀ 01 12 nM 162 nM 43370 nM 02 33 nM 1139 nM >49992 nM 03 25 nM 1022 nM 23641 nM 04 49 nM 806 nM >50000 nM 05 73 nM 629 nM >50000 nM 06 61 nM 593 nM >50000 nM 07 37 nM 531 nM >50000 nM 08 37 nM 524 nM >50000 nM 09 39 nM 489 nM 6174 nM 10 52 nM 407 nM >50000 nM AND 12 nM 401 nM >49954 nM 12 38 nM 385 nM >49980 nM 13 43 nM 358 nM >50000 nM 14 41 nM 306 nM 14255 nM 15 38 nM 263 nM >50000 nM 16 30 nM 262 nM >50000 nM 17 8 nM 251 nM >50000 nM 18 37 nM 244 nM >50000 nM 19 32 nM 214 nM >50000 nM 20 54 nM 192 nM >50000 nM 21 20 nM 190 nM >49974 nM 22 And nM 188 nM >50000 nM 23 62 nM 180 nM >50000 nM 24 28 nM 165 nM >50000 nM 25 36 nM 164 nM 26661 nM 26 45 nM 164 nM >50000 nM 27 51 nM 160 nM >50000 nM 28 42 nM 158 nM >49948 nM 29 36 nM 151 nM >49966 nM

- 15 043735

thirty 46 nM 126 nM 11387 nM 31 21 nM 121 nM >49970 nM 32 17 nM 73 nM 39500 nM 33 15 nM 49 nM 33032 nM 34 14 nM 14 nM 15847 nM 35 38 nM 207 nM >50000 nM 36 67 nM 551 nM >50000 nM 37 15 nM 451 nM 22572 nM 38 13 nM 278 nM >49976 nM 39 19 nM 262 nM 16975 nM 40 26 nM 125 nM >50000 nM 41 5 nM 123 nM 25390 nM 42 20 nM 87 nM >49973 nM 43 16 nM 69 nM 36481 nM 44 14 nM 574 nM >50000 nM 45 10 nM 307 nM 11399 nM 46 10 nM 234 nM >49993 nM 47 5 nM 144 nM 23169 nM 48 24 nM 67 nM 1485 nM 49 21 nM 50 nM >50000 nM 50 20 nM 24 nM >50000 nM 51 13 nM 325 nM 48005 nM 52 9 nM 315 nM 41750 nM 53 15 nM 19 nM >50000 nM 54 308 nM 4 nM >50000 nM 55 391 nM 50 nM >49957 nM 56 89 nM 34 nM 34674 nM 57 2690 nM 6 nM >50000 nM 58 114 nM 363 nM >49945 nM 59 18 nM But. 27250 nM 60 21 nM 92 nM >50000 nM 61 26 nM 61 nM >50000 nM 62 18 nM But. >50000 nM 63 19 nM But. >50000 nM 64 17 nM But. >50000 nM 65 And nM But. >50000 nM 66 23 nM But. 31003 nM

According to AOC2 tissue enzymatic activity, the only human tissue with high AOC2-like activity is the retina, and the expression is associated with retinal capillaries as shown by immunohistochemical studies. According to the enzymatic function and localization of expression, the physiological function of AOC2 may resemble the homologue AOC3, which has been described as being relevant for example in neurovascular inflammation, retinal inflammation and immune cell recruitment (Matsuda et al. Invest Ophthalmol Vis Sci. 2017, 58(7): 3254-3261, Noda et al. FASEB J. 2008, 4: 1094-103). Data on pharmacological inhibition or genetic depletion of AOC2

- 16 043735 is not yet available, and therefore it is difficult to assess the contribution of AOC2 to inflammation of the retina and blood vessels.

However, compared to AOC3 inhibition alone, combined inhibition of AOC2 and AOC3 may enhance anti-inflammatory effects in humans, particularly in the treatment of ocular diseases.

Therefore, the object of the invention was to provide compounds with high activity against AOC3 and AOC2 to achieve the desired pharmacological effects.

It has now been found that, surprisingly, the compounds of the present invention are more active AOC2 inhibitors than the corresponding compounds from the prior art, such as those described in WO 2013/163675 and WO 2018/027892, i.e. replacing the phenyl moiety with a pyridinyl moiety and introducing azetidinyl, pyrrolidinyl or piperidinyl sulfonyl amides results in compounds with improved AOC2 inhibitory activity without affecting AOC3 activity.

Since it has a secondary amine substituent in the sulfonamide group, compound 14 of WO 2013/163675 represents the closest comparative compound in structure to the cyclic amines claimed in the present invention at the same position. Compound 14 of WO 2013/163675 contains a dimethylaminosulfonamide group compared to the cyclic azetidinyl, pyrrolidinyl or piperidinyl sulfonamides disclosed in the present invention. In addition, compound 14 of WO 2013/163675 contains a phenyl group, while the compounds disclosed in the present invention contain a pyridinyl group. Although compound 14 of WO 2013/163675 is a weak AOC2 inhibitor (IC ₅₀ = 1164 nM, approximately 145 times higher than the IC ₅₀ for AOC3), the compound of the present invention exhibits improved AOC2 inhibitory activity as shown in Examples 42, 35, 40 (each approximately 5 times less active on AOC2 compared to AOC3) and 45 (each approximately 30 times less active on AOC2 compared to AOC3) in the following table.

Reference compounds A and B, which differ structurally from Examples 42 and 35 according to the present invention only by a phenyl group compared to a pyridinyl group, can be prepared analogously to the synthesis described in WO 2013/163675. In comparison, the pyridinyl derivatives of the present invention exhibit enhanced AOC2 inhibitory capacity. Control compound A is 22 times less active (ratio IC ₅₀ AOC2 / IC ₅₀ AOC3) against AOC2 compared to AOC3, while the pyridinyl analogue from Example 42 is only 4 times less active against AOC2. Control B is 92 times less active against AOC2 than AOC3, while the pyridinyl analogue of Example 42 is only 5 times less active against AOC2.

AOC1 expression and enzymatic activity are mainly found in the intestine, placenta, and kidney. The enzyme catalyzes the oxidation of primary amines obtained from food and protects humans from the cardiometabolic effects of histamine, putrescine, tryptamine and cadaverine. Inhibition of AOS1 may lead to impaired tolerance to oral histamine, leading to increased plasma and tissue histamine levels, which may cause adverse events or undesirable side effects such as decreased blood pressure and compensation by increased heart rate, tachycardia, headache pain, hot flashes, urticaria, itching, bronchospasm and cardiac arrest (Maintz L. and Novak N. 2007. Am. J. Clin. Nutr. 85: 1185-96). The consequences of AOC1 inhibition in combination with histamine intake were demonstrated in experiments on pigs: after administration of the AOC1 inhibitor aminoguanidine (100 mg/kg) and histamine probing (2 mg/kg), an increase in the level of histamine in the blood was observed in animals, accompanied by a drop in blood pressure, increased pulse, flushing, vomiting and death (3 of 15 animals) (Sattler J. 1988. Agents and Actions, 23: 361-365) under experimental conditions. Histamine intolerance in humans has been associated with mutations in the AOC1 promoter region, resulting in decreased plasma AOC1 mRNA expression and activity (Maintz et al. 2011. Allergy 66: 893-902).

Therefore, the object of the invention was to provide compounds with low activity against AOC1 in order to avoid such undesirable side effects.

It has now been found that, surprisingly, the compounds of the present invention exhibit increased selectivity for AOC1 compared to prior art compounds, in particular the compounds disclosed in WO 2018/027892. Examples 6, 5 and 2 from WO 2018/027892 differ from examples 35, 40 and 45, respectively, by a pyrimidinyl group compared to a pyridinyl group and the absence of a sulfonyl group. While Example 6 of WO 2018/027892 and the pyridinyl sulfonyl analogue of Example 35 of the present invention are equally effective against AOC3, Example 35 shows a much higher IC50 against AOC1. Example 5 of WO 2018/027892 and the racemic pyridinyl sulfonyl analogue of Example 40 of the present invention are equally effective against AOC3, however Example 40 shows a much higher IC50 against AOC1. In addition, example 2 of WO 2018/027892 and the pyridinylsulfonyl analogue of example 45 in accordance with the present invention are equally effective against AOC3, however,

- 17 043735 example 45 shows a much higher IC ₅₀ in relation to AOC1.

Comparison of biological data of some compounds obtained in the AOC3, AOC2 and

AOC1 as described above.

Structure IC ₅₀ AOSZ 1C ₅₀ AOS2 IC ₅₀ AOC1 Oh Oh Connection 14 from WO 2013/163675 8 nM 1164 nM >50000 nM ^F ABOUT Comparative compound A 13 nM 287 nM >49978 µM ^F > _B XJ O O Example 42 20 nM 87 nM >49973 nM

_X F but if O ^x o Comparative compound B 27 nM 2515 nM >50000 mkM .F but if 'd /Ν. ,0. Α,ΝΗ ₂ α,υ 0^*0 Example 35 38 nM 207 nM >50000 mkM ^F _n ^o^Cnh ₂ HO/ ^N Example 6 from WO 2018/027892 59 nM 1118 nM 1171 nM .F _ν ^°Α/νΗ ₂ /-l/ Ο··/ NN Example 5 from WO 2018/027892 13 nM 497 nM 268 nM ,F —Ο ί| \__ ,NO^^^NH ₂ α _5ί α ^x o Example 40 26 nM 125 nM >50000 mkM .F _ν ^ο^//νη ₂ H Example 2 from WO 2018/027892 20 nM 1085 nM 269 nM .F ^HO \/\ V. JU c/ ^s< o Example 45 10 nM 307 nM 11399 nM

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In view of their ability to inhibit AOC3 and AOC2, the compounds of general formula (I) according to the invention and their corresponding salts are suitable for the treatment, including prophylactic treatment, of all those diseases or conditions that may be affected by or which are mediated by inhibition of the activity of AOC3 and AOC2.

In addition, the compounds of the present invention exhibit moderate to high in vitro efflux and/or low intrinsic permeability in the MDCK p-GP assay. Therefore, the compounds of the present invention are expected to exhibit lower free concentrations in the brain than in the blood (Liu, H. et al., 2018, Drug Discovery Today 23 (7): 1357-1372).

Accordingly, the present invention relates to a compound of general formula (I) as a drug.

Furthermore, the present invention relates to the use of a compound of general formula (I) for the treatment and/or prevention of diseases or conditions that are mediated by inhibition of AOC3 in a patient, preferably a human.

In yet another aspect, the present invention provides a method of treating, including preventing a disease or condition mediated by AOC3 inhibition in a mammal, which includes the step of administering to a patient, preferably a human, in need of such treatment a therapeutically effective amount of a compound of the present invention or a pharmaceutical composition thereof.

Diseases and conditions mediated by AOC3 inhibitors include cancer, NASH (non-alcoholic steatohepatitis), pulmonary fibrosis, retinopathy, nephropathy and stroke.

In one aspect, the compounds of the present invention are particularly useful for the treatment of inflammatory diseases such as inflammatory vascular diseases, arthritis, acute and chronic joint inflammation; eczema such as atopic eczema, ulcerative psoriasis and rheumatoid psoriasis; pain, especially musculoskeletal or nociceptive pain; inflammatory bowel disease, especially non-infectious inflammatory bowel disease; multiple sclerosis; scleroderma, pulmonary diseases such as respiratory distress syndrome, asthma, pulmonary fibrosis, idiopathic pulmonary fibrosis (IPF), chronic obstructive pulmonary disease (COPD) and idiopathic inflammatory disease; nephropathy, diabetic proteinuria, renal fibrosis; diabetic retinopathy or diabetic edema, such as diabetic macular edema; cancer, in particular melanoma and lymphoma; hepatocellular carcinoma, nonspecific colitis, rheumatoid arthritis, Crohn's disease, colitis, biliary tract diseases, primary biliary cholangitis, primary sclerosing cholangitis, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), alcoholic liver disease, liver fibrosis, liver cirrhosis; ulcerative reperfusion injury, cerebral ischemia and graft rejection.

According to another aspect, the compounds of the present invention are particularly useful for the treatment of inflammatory diseases such as inflammatory vascular diseases, arthritis and inflammatory bowel disease, especially non-infectious inflammatory bowel disease; pulmonary fibrosis and idiopathic pulmonary fibrosis; ; diabetic retinopathy or diabetic edema, such as diabetic macular edema; nonspecific colitis, rheumatoid arthritis, Crohn's disease, colitis; biliary tract diseases, primary biliary cholangitis, primary sclerosing cholangitis, non-alcoholic steatohepatitis (NASH), non-alcoholic fatty liver disease (NAFLD), alcoholic liver disease, liver fibrosis and cirrhosis.

The dosage range of the compounds of general formula (I) applicable per day is generally from 0.001 to 10 mg per kg of patient's body weight, preferably from 0.01 to 8 mg per kg of patient's body weight. Each dosage unit may typically contain from 0.1 to 1000 mg of active substance, preferably from 0.5 to 500 mg of active substance.

Of course, the actual pharmaceutically effective amount or therapeutic dose will depend on factors known to those skilled in the art, such as the age and weight of the patient, the route of administration, and the severity of the disease. In any case, the combination should be administered in doses and by a route that delivers a pharmaceutically effective amount in accordance with the particular patient's condition.

Pharmaceutical compositions.

Suitable preparations for administering the compounds of formula (I) will be apparent to those skilled in the art and include, for example, tablets, pills, capsules, suppositories, lozenges, lozenges, solutions, syrups, elixirs, sachets, injectables, inhalants, powders, etc. The content of pharmaceutically active compound(s) is preferably in the range from 0.1 to 90, for example from 1 to 70 wt.% of the composition as a whole.

Suitable tablets can be prepared, for example, by mixing one or more compounds of formula (I) with known excipients, for example, inert diluents, carriers, disintegrants, adjuvants, surfactants, binders and/or lubricants. Tablets can also consist of several layers.

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Combination therapy.

The compounds of the invention may be further combined with one or more, preferably one, additional therapeutic agents. In one embodiment, the additional therapeutic agent is selected from a group of therapeutic agents useful in the treatment of diseases or conditions associated with metabolic syndrome, diabetes, obesity, cardiovascular disease, NASH (nonalcoholic steatohepatitis) cancer, pulmonary fibrosis, retinopathy, nephropathy, and/or stroke.

Therefore, the compound according to the invention can be combined with one or more additional therapeutic agents selected from the group consisting of anti-obesity agents (including appetite suppressants), blood glucose-lowering agents, antidiabetic agents, agents for the treatment of dyslipidemias such as lipid-lowering agents, antihypertensive agents, anti-atherosclerotic agents, anti-inflammatory active substances, anti-fibrotic agents, anti-cancer agents, antithrombotic agents, anti-angiogenesis agents, agents for the treatment of heart failure and agents for the treatment of complications caused by or associated with diabetes.

Preferably, the compounds of the present invention and/or pharmaceutical compositions containing a compound of the present invention, optionally in combination with one or more additional therapeutic agents, are administered in conjunction with exercise and/or diet.

Therefore, in another aspect, the present invention relates to the use of a compound according to the invention in combination with one or more additional therapeutic agents described above and below, for the treatment or prevention of diseases or conditions that may be affected by or that are mediated by inhibition of AOC3, in particular , diseases or conditions described above and below.

In yet another aspect, the present invention provides a method of treating, including preventing, a disease or condition mediated by inhibition of AOC3 in a patient, said method comprising the step of administering to the patient, preferably a human in need of such treatment, a therapeutically effective amount of a compound according to the present invention. the invention in combination with a therapeutically effective amount of one or more additional therapeutic agents described above and below.

The use of a compound according to the invention in combination with an additional therapeutic agent can occur simultaneously or at intervals.

The compound according to the invention and one or more additional therapeutic agents can both be present together in one composition or separately in two identical or different compositions, for example, in the form of a so-called kit of components.

Therefore, in another aspect, the present invention relates to a pharmaceutical composition that includes a compound according to the invention and one or more additional therapeutic agents described above and below in the present invention, optionally together with one or more inert carriers and/or diluents.

Synthesis schemes.

Typical methods for preparing the compounds of the invention are described in the experimental section.

The potent inhibitory effect of the compounds according to the invention can be determined using in vitro enzyme assays as described in the experimental section.

The compounds of the present invention can also be prepared by methods known in the art, including the methods described below and including variations available to those skilled in the art.

Scheme 1

Compounds of general formula I, in which A and R ¹ have the previously defined definitions, can be prepared using the method described in Scheme 1 using a compound of general formula 1-1. Deprotection of the tert-butoxycarbonyl (=BOC) group can be accomplished by treatment with an acid such as hydrochloric acid or trifluoroacetic acid in a suitable solvent such as methanol, dioxane or dichloromethane at a temperature of -20 to 100°C. When 1-1 is used as a mixture of E/Z isomers, the vinyl fluoride E/Z isomers of the compounds of general formula I can be separated by preparative HPLC or silica gel column chromatography, which gives the compounds of general formula I in isomerically pure form.

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Scheme 2

Intermediates of general formula 1-1, in which A and R ¹ have the previously defined definitions, can be prepared using the method described in Scheme 2 using a 6-fluoro-6-chloro-pyridinylsulfonamide compound of general formula 2-1, in which A and R ¹ are as previously defined, and the alcohol 2-2 either as the pure E isomer or as an E/Z mixture, and a base such as sodium t-butoxide or sodium hydride in a suitable solvent such as THF, DMSO or toluene at temperatures from -20 to 100°C.

Scheme 3

3-1 3-2 2-1

Intermediates of general formula 2-1, in which A and R ¹ are as previously defined, can be prepared using the method described in Scheme 3 using an amino compound of general formula 3-1, in which A and R ¹ are as previously defined, and 6-fluoro- or 6-chloropyridine-3-sulfonyl chloride and a base such as triethylamine in a suitable solvent such as dichloromethane, NMP, THF, DMSO or mixtures thereof at -20 to 100°C.

Scheme 4

4-1 4-2 3-1 a

Intermediates of general formula 3-1a, in which the amine substituents R are selected as previously defined for amides from the substituent R ¹ , can be prepared by the method described in Scheme 4 using a carboxylic acid of general formula 4-1, a primary or secondary amine general formula 4-2, wherein the amine substituents R are selected as previously defined for amides from the substituent R ¹ , an amide coupling reagent such as 1-propanephosphonic acid cyclic anhydride or HATU, and a base such as triethylamine or DIPEA in a suitable solvent, such as THF or DMF at temperatures from -20 to 100°C.

Scheme 5

Compounds of the general formula 1-amide, which have an amide group according to the definitions for R ¹ , can also be prepared from carboxylic acids of general formula 5-1, a primary or secondary amine of general formula 4-2, where the amine substituents R are chosen as previously defined for amides from the R ¹ substituent, a reagent coupling of an amide such as 1-propanephosphonic acid cyclic anhydride, TCFH or HATU, and a base such as triethylamine or DIPEA in a suitable solvent such as THF or DMF at -20 to 100° WITH. Carboxylic acids of general formula 5-1 are available from the corresponding alkyl esters by saponification with sodium or lithium hydroxide in a solvent such as methanol or THF at -20 to 100°C.

The presented synthesis routes can be based on the use of protecting groups. For example, reactive groups present, such as hydroxy, carbonyl, carboxy, amino, alkylamino or imino, can be protected during the reaction by conventional protecting groups, which are cleaved off again after the reaction. Suitable protecting groups for the respective functional groups and their removal are well known to those skilled in the art and are described in the organic synthesis literature.

The compounds of general formula I can be separated into their enantiomers and/or diastereomers, as mentioned previously.

Compounds of the general formula I, which occur as racemates, can be separated by known methods into their optical antipodes, and diastereomeric mixtures of compounds of the general formula I can be separated into their diastereomers, using their different physicochemical properties with use as such known methods, for example, chromatography and/or fractional crystallization; if the resulting compounds are racemates, they can be separated into enantiomers as follows.

Racemates are preferably separated by column chromatography on chiral phases or by crystallization from an optically active solvent or by reaction with an optically active substance that forms salts or derivatives, such as esters or amides, with the racemic compound. Salts can be formed with enantiomerically pure acids for basic compounds and with enantiomerically pure bases for acidic compounds.

Diastereomeric derivatives are formed with enantiomerically pure auxiliary compounds, such as acids, their activated derivatives or alcohols. Separation of the diastereomeric mixture of salts or derivatives thus obtained can be achieved by using their different physicochemical properties, for example, differences in solubility; free antipodes can be released from pure diastereomeric salts or derivatives by the action of suitable agents. Optically active acids commonly used for this purpose, as well as optically active alcohols useful as auxiliary residues, are known to those skilled in the art.

As indicated above, the compounds of formula I can be converted into salts, in particular for pharmaceutical use, into pharmaceutically acceptable salts. As used herein, the term pharmaceutically acceptable salts refers to derivatives of the disclosed compounds in which the parent compound is modified to form acidic or basic salts thereof.

Experimental section.

The following examples are intended to illustrate the present invention without limiting it.

General definitions.

List of abbreviations.

A acid

ACN Acetonitrile aq. water

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V base VOS tert-butoxycarbonyl °C degree Celsius Cbz benzioloxycarbonyl d day DXM dichloromethane DIPEA N,N-diisopropylethylamine DMF N,N-dimethylformamide DMSO dimethyl sulfoxide eq. equivalent ERI-MS electrospray ionization mass spectrometry EtOH ethanol EtOAc ethyl acetate hut excess G gram h hour HATU X,X,X',X'-tetramethyl-O-(7azabenzotriazol-1-yl)uronium hexafluorophosphate HPLC high performance liquid chromatography IBCF isobutyl chloroformate iPrOH isopropyl alcohol l liter M molar (mol/l) MeOH methanol min minute mg milligram ml milliliter mmol millimoles MS mass spectrometry MTBE 2-methoxy-2-methylpropane N normal = 1 molar = 1 mol/l NMP X-methyl-2-pyrrolidinone NMR nuclear magnetic resonance

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Pd/C palladium on coal psi inch lbf per square inch OF reverse phase CT room temperature (approx. 22 °C) By S retention time solvent us. saturated

T temperature t time TBTU benzotriazolyltetramethyluronium tetrafluoroborate TCFH chloro-Ν,Ν,Ν' ,Ν' tetramethylformamidinium hexafluorophosphate TLC thin layer chromatography TEA triethylamine TFU trifluoroacetic acid THF tetrahydrofuran TNR tetrahydropyran Tol General methods. toluene

Unless otherwise stated, all reactions are carried out at room temperature (about 22°C), under an inert atmosphere (eg, argon, N ₂ ) under anhydrous conditions. All compounds are characterized by at least one of the following methods: 1H NMR, HPLC, HPLC-MS or melting point.

Typically, the progress of the reaction is monitored using thin layer chromatography (TLC) or HPLC-MS. Intermediates and products are purified using at least one of the following methods:

Recrystallization, silica gel column chromatography or reverse phase HPLC using a semipreparative C18 column, eluting with a gradient:

ACN and H ₂ O + 0.1% TFA

ACN and H ₂ O + 0.1% NH ₄ OH

Analytical data.

The mass spectrometry (MS) data shown correspond to the observed mass signals (e.g., [M+H]+). The HPLC methods used to characterize the compounds of the invention are described in the following tables.

HPLC methods.

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Method Mobile phase A Mobile phase B Gradient Flow (ml/min) Column Temperature HPLC-1 0.1% TFA in water ACN Time (min) %A %IN HB ridge VEN C18_2.1 x 30 mm_1.7 µM particle diameter 60°С 0.0 99.0 GO 1.6 0.02 99.0 th 1.6 GO 0.0 100.0 1.6 1.1 0.0 100.0 1.6

Method Mobile phase A Mobile phase B Gradient Flow (ml/min) Column Temperature HPLC-2 0.1% TFA in water ACN Time (min) %A %IN Sunfire Cl 8-2.1 x 30 MM-2.5 µM particle diameter 60°С 0.0 99.0 1.0 1.5 0.02 99.0 1.0 1.5 1.0 0.0 100.0 1.5 1.1 0.0 100.0 1.5

Method Under- visual phase A Mobile phase B Gradient Flow (ml/min) Column Temperature HPLC-3 0.1% TFA in water ACN Time (min) %A %V Sunfire C 18-2.1 x 30 mm-2.5 µm particle diameter 60°С 0.0 50.0 50.0 1.5 0.02 50.0 50.0 1.5 1.0 0.0 100.0 1.5 1.1 0.0 100.0 1.5

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Method Mobile phase A Mobile phase B Gradient Flow (ml/min) Column Temperature HPLC-4 0.1% NH ₃ in water ACN Time (min) %A %IN XB ridge C18_3.0 x 30 mm_2.5 µM particle diameter 60°С 0.0 97.0 3.0 2.2 0.2 97.0 3.0 2.2 1.2 0.0 100.0 2.2 1.25 0.0 100.0 3.0 1.4 0.0 100.0 3.0

Method Mobile phase A Mobile phase B Gradient Flow (ml/min) Column Temperature HPLC-5 0.1% TFA in water ACN Time (min) %A %IN XB ridge C18_3.0 x 30 mm_2.5 µM particle diameter 60°С 0.0 97.0 3.0 2.2 0.2 97.0 3.0 2.2 1.2 0.0 100.0 2.2 1.25 0.0 100.0 3.0 1.4 0.0 100.0 3.0

Method Mobile phase A Mobile phase B Gradient Flow (ml/min) Column Temperature HPLC-6 0.1% TFA in water ACN Time (min) %A %IN Sunfire C18_3.0 x 30 mm_2.5 µM particle diameter 60°С 0.0 97.0 3.0 2.2 0.2 97.0 3.0 2.2 1.2 0.0 100.0 2.2 1.25 0.0 100.0 3.0 1.4 0.0 100.0 3.0

Method Mobile phase A Mobile phase B Gradient Flow (ml/min) Column Temperature HPLC-7 0.1% TFA in water 0.08% TFU in ACN Time (min) %A %V Sunfire C18_3.0 x 30 mm_2.5 µM 60°С 0.0 95.0 5.0 1.5 1.3 0.0 100.0 1.5 1.5 0.0 100.0 1.5

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1.6 95.0 5.0 1.5 particle diameter

Method Mobile phase A Mobile phase B Gradient Flow (ml/min) Column Temperature HPLC-8 0.1% NH ₃ in water ACN Time (min) %A %IN XB ridge C18_3.0 x 30 mm_2.5 µM particle diameter 60°С 0.0 95.0 5.0 1.5 1.3 0.0 100.0 1.5 1.5 0.0 100.0 1.5 1.6 95.0 5.0 1.5

Method Mobile phase A Mobile phase B Gradient Flow (ml/min) Column Temperature HPLC-9 0.1% TFA in water 0.08% TFU in ACN Time (min) %A %IN Sunfire C 18-3.0 x 30 mm_2.5 µM particle diameter 60°С 0.0 95.0 5.0 1.5 1.3 0.0 100.0 1.5 1.5 0.0 100.0 1.5 1.6 95.0 5.0 1.5

Method Mobile phase A Mobile phase B Gradient Flow (ml/min) Column Temperature HPLC- 10 0.1% TFA in water ACN Time (min) %A %IN Sunfire C 18-3.0 x 30 mm-2.5 µm particle diameter 60°С 0.0 97.0 3.0 4.0 0.15 97.0 3.0 3.0 2.15 0.0 100.0 3.0 2.2 0.0 100.0 4.5 2.4 0.0 100.0 4.5

Synthetic intermediates/examples.

The intermediates and examples that follow are illustrative, and, as one skilled in the art will recognize, specific reagents or conditions can be varied as needed for individual compounds without undue experimentation.

The compounds of the invention can be prepared by the general methods and examples presented below and by methods known to those skilled in the art. Optimal reaction conditions and reaction times may vary depending on the specific reagents used. Unless otherwise indicated, solvents, temperatures, pressures and other reaction conditions can be easily selected by one skilled in the art. Specific procedures are presented in the synthesis section. Undescribed intermediates used in the following syntheses are either commercially available or can be easily prepared by methods known to those skilled in the art. The progress of the reaction can be monitored by conventional methods such as thin layer chromatography (TLC) or high pressure liquid chromatography-mass spectrometry (HPLC-MS). Intermediates and products can be purified by methods known in the art, including column chromatography, HPLC, preparative TLC or recrystallization.

Carboxylic acid intermediate.

Stage 1 connection I.1. Trans-3-aza-bicyclo[3.1.0]hexane-6 methylamide hydrochloride

amide combination. To a solution of trans-3-aza bicyclo[3.1.0]hexane-3,6-dicarboxylic acid 3-tert-butyl ester (1.00 g; 4.40 mmol) and TEA (4.94 ml; 35.20 mmol) in THF (5 ml) was added methylamine (2M in THF; 4.40 ml; 8.80 mmol). The reaction mixture was stirred at RT for 5 min. and 1-propanephosphonic acid cyclic anhydride (50% in THF; 5.14 mL; 8.80 mmol) was added. The reaction mixture was stirred at RT for 45 minutes, diluted with aq. 4 N NaOH (25 ml) and extracted with MTBE (2x25 ml). The combined organic phases were dried with Na ₂ SO ₄ , filtered and evaporated to dryness.

Stage 2 - removal of BOS protection. The crude material from step 1 was resuspended in EtOAc (20 ml) and MeOH (20 ml) and hydrogen chloride (4 N in 1,4-dioxane; 5 ml; 20.00 mmol) were added. Reaction mixture

- 27 043735 was stirred at RT overnight and concentrated under reduced pressure to give intermediate 1.1.

Yield: 882 mg (80%), ESI-MS: m/z = 141 [M+H] ⁺ , By (HPLC): 0.12 min (HPLC-6).

Intermediate I.2. Trans-3-aza-bicyclo[3.1.0]hexane-1carboxylic acid methylamide hydrochloride. Amide combination.

n°^ <0^° / ^{+ /NH2} ” <5%^° / n G ⁷

1.2a

Trans-3-aza-bicyclo[3.1.0]hexane-1,3-dicarboxylic acid 3-tert-butyl ester (1.00 g, 4.40 mmol) and HATU (1.90 g, 4.84 mmol) were dissolved in DMF (5 ml) and DIPEA (1.89 ml; 11.00 mmol) and stirred at RT for 30 min. Methylamine (2M in THF; 4.40 mL; 8.80 mmol) was added to the reaction mixture and stirred at RT overnight. The reaction mixture was diluted with water (20 ml) and extracted with DCM (3x20 ml). The combined organic phases were washed with aq. 1 N NaOH, dried and concentrated under reduced pressure. The residue was purified by RP-HPLC (ACN/water+TFA) to give intermediate I.2a.

Yield: 0.95 g (90%), ESI-MS: m/z = 185 [M+H] ⁺ , By (HPLC): 0.87 min (HPLC-6).

Removing BOS protection.

Intermediate I.2a (0.94 mg; 3.89 mmol) was dissolved in MeOH (2 mL) and hydrogen chloride (4 N in 1,4-dioxane; 5.00 mL; 20.00 mmol) was added. The reaction mixture was stirred at RT for 1 hour 40 minutes, then reduced in vacuo and co-evaporated with MeOH to give intermediate I.2.

Yield: 0.65 g (95%), ESI-MS: m/z = 191 [M+H] ⁺ , By (HPLC): 0.09 min (HPLC-10).

Intermediate I.3. [2-Methyl-2-(tetrahydro-pyran-2-yloxy)propyl]amide (S)pyrrolidine-3-carboxylic acid. Amide combination.

(S)-Pyrrolidine-1,3-dicarboxylic acid 1-benzyl ester (2.00 g; 8.02 mmol) was dissolved in THF (20.00 mL) and TEA (9.01 mL; 64.19 mmol) and 1-amino-2-methylpropane-2 was added -ol (0.83 g; 8.83 mmol). The reaction mixture was cooled to 0°C and a solution of 1-propanephosphonic acid cyclic anhydride (50% in THF; 7.03 ml; 12.04 mmol) was added. It was stirred at RT for 3 hours. The reaction mixture was diluted with aq. 4 N NaOH (20 ml) and extracted twice with MTBE (30 ml). The combined organic phases were dried and evaporated to give crude intermediate I.3a.

Yield: 2.51 g (98%), ESI-MS: m/z = 321 [M+H] ⁺ , Wu (HPLC): 0.90 min (HPLC-6).

Cbz protection removal.

A mixture of intermediate X.3a (2.51 g, 7.83 mmol) and 10% Pd/C (0.25 g) in MeOH (50 ml) was treated with hydrogen (50 psi) at RT overnight. The reaction mixture was filtered, washed with MeOH and concentrated in vacuo to give crude intermediate I.3b.

Yield: 1.47 g (99%), ESI-MS: m/z = 187 [M+H] ⁺ , Wu (HPLC): 0.12 min (HPLC-6).

TNR protection.

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Intermediate I.3b (1.47 g, 7.89 mmol) was diluted with 3,4-dihydro-2H-pyran (10.00 mL, 108.28 mmol) and p-toluenesulfonic acid monohydrate (0.15 g, 0.79) was added mmol). The reaction mixture was stirred at RT for three days and concentrated in vacuo to give crude intermediate 1.3.

Yield: 2.54 g (99%), ESI-MS: m/z = 271 [M+H] ⁺ , Wu (HPLC): 0.75 min (HPLC-4)

Intermediate I.4. Trifluoroacetate (8)-X-piperidin-3-yl-acetamide.

(S)-3-acetylamino-piperidine-1-carboxylic acid tert-butyl ester (3.00 g; 12.38 mmol), trifluoroacetic acid (9.54 mL; 123.80 mmol) and DCM (80 mL) were stirred at RT overnight. The reaction mixture was evaporated and co-evaporated with EtOH twice to give intermediate 1.4.

Yield: 4.20 g (quantitative), ESI-MS: m/z = 143 [M+H]+.

Intermediate I.5. Morpholin-4-yl-(4-phenyl-piperidin-4-yl)methanone.

Intermediate I.5 can be prepared according to the procedure described in WO 98/27086, cc. 38-40. The starting materials were morpholine and 4-phenyl-piperidine-1,4-dicarboxylic acid mono-tert-butyl ester.

Intermediate I.6. Azetidin-1-yl-piperidin-4-yl-methanone trifluoroacetate. Amide combination.

Piperidine-1,4-dicarboxylic acid mono-tert-butyl ester (2.00 g; 8.72 mmol), TBTU (2.89 g; 9.00 mmol) and TEA (1.25 ml; 9.00 mmol) were dissolved in THF and stirred at RT for 1 h Azetidine (0.61 ml; 9.00 mmol) and TEA (1.25 ml; 9.00 mmol) were added to the reaction mixture. The reaction mixture was stirred at RT overnight, diluted with water and extracted with EtOAc. The combined organic phases were dried with Na ₂ SO ₄ and reduced in vacuo to give crude intermediate I.6a.

Yield: 2.00 g (85%), ESI-MS: m/z = 269 [M+H]+.

Removing BOS protection.

Intermediate I.6a (2.00 g; 7.45 mmol) was dissolved in DCM (20 mL) and TFA (2.23 mL; 30.00 mmol) was added. The reaction mixture was stirred at RT overnight and reduced in vacuo. The residue was resuspended in DCM, filtered through an HCO ₃ cartridge and the filtrate was evaporated under reduced pressure to give intermediate 1.6. Yield: 2.80 g (quantit.), ESI-MS: m/z = 169 [M+H] ⁺ .

I.7. 1-(3-Piperidin-4-yl-azetidin-1-yl)ethanone.

Cbz protection.

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3-Piperidin-4-yl-azetidin-1-carboxylic acid tert-butyl ester (500 mg; 2.08 mmol) was dissolved in DCM (10 mg), treated with TEA (348 μl; 2.50 mmol) and cooled to 0°C. Benzyl chloroformate (322 μL; 2.29 mmol) was added dropwise to the reaction mixture, and the reaction mixture was then heated to RT. The reaction mixture was stirred at RT overnight, diluted with DCM and extracted twice with water. The organic phase was dried and concentrated in vacuo. The crude material was purified by silica gel chromatography (cyclohexane/EtOAC) to give intermediate I.7a.

Yield: 220 mg (28%), ESI-MS: m/z = 375 [M+H]+, By (HPLC): 0.81 min (HPLC-2).

Removing BOS protection.

To a solution of intermediate I.7a (220 mg; 0.59 mmol) in DCM (3 mL) was added TFA (453 μL; 5.87 mmol). The reaction mixture was stirred at RT overnight, the solvent was evaporated under reduced pressure, and the residue was washed once with water and once with aq. NaHCO ₃ solution. The organic phase was dried and concentrated in vacuo to give crude intermediate I.7b.

Yield: 170 mg (100%), ESI-MS: m/z = 275 [M+H] ⁺ , By (HPLC): 0.43 min (HPLC-2).

Acetylation.

Intermediate I.7b (170 mg; 0.62 mmol) was dissolved in DCM (3.00 mL) and treated with TEA (258 μL; 1.86 mmol). The solution was cooled to 0°C and acetyl chloride (53 µl; 0.74 mmol) was added dropwise. The reaction mixture was stirred at 0°C for 10 min, warmed to RT and stirred at RT overnight. The reaction mixture was washed with water twice. The organic phase was dried and concentrated in vacuo to give crude intermediate I.7c.

Yield: 190 mg (97%), ESI-MS: m/z = 317 [M+H] ⁺ , By (HPLC): 0.60 min (HPLC-2).

Cbz protection removal.

A mixture of intermediate I.7c (190 mg; 0.60 mmol) and 10% Pd/C (50 mg) in MeOH (5 ml) was treated with hydrogen (50 psi) at RT overnight. The reaction mixture was filtered and concentrated in vacuo to give crude intermediate I.7.

Yield: 90 mg (82%), ESI-MS: m/z = 183 [M+H] ⁺ .

Intermediate I.8. N,N-dimethyl-2-piperidin-4-yl-acetamide trifluoroacetate.

Intermediate I.8 was prepared according to the procedure described in WO 2008/071646, cc. 81-82.

I.9. N-ethyl-2-piperidin-4-yl-acetamide hydrochloride. Amide combination.

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4-Carboxymethyl-piperidine-1-carboxylic acid tert-butyl ester (3.00 g; 12.33 mmol), TBTU (3.96 g; 12.33 mmol) and TEA (5.19 mL; 36.99 mmol) were dissolved in DMF (10 mL). The solution was stirred at RT for 10 min. Ethylamine hydrochloride (1.01 g, 12.33 mmol) was added to the reaction mixture and it was stirred at RT overnight. TBTU was added to the reaction mixture and after 5 min. Ethylamine hydrochloride (0.5 g, 6.15 mmol) was added while stirring at RT. After stirring for 4 hours at RT, the reaction mixture was extracted with EtOAc. The organic phases were concentrated in vacuo. The crude material was dissolved in DCM, filtered through an Alox main cartridge, and the filtrate was washed with aq. 0.1 N HCl and evaporated under reduced pressure to obtain intermediate I.9a.

Yield: 3.3 g (99%), ESI-MS: m/z = 271 [M+H]+, By (HPLC): 0.75 min (HPLC-4).

Removing BOS protection.

Intermediate I.9a (3.30 g; 12.21 mmol) was dissolved in 1,4-dioxane (30 ml) and a solution of 4 N hydrogen chloride in 1,4-dioxane (6.10 ml; 24.41 mmol) was added. The reaction mixture was stirred at RT overnight. A solution of 4 N hydrogen chloride in 1,4 dioxane (6.10 ml; 24.41 mmol) was added to the reaction mixture and it was stirred at RT overnight. The reaction mixture was diluted with diethyl ether and the precipitate was filtered to give intermediate I.9.

Yield: 2.52 g (100%), ESI-MS: m/z = 171 [M+H]+, By (HPLC): 0.78 min (HPLC-6)

Intermediate I.10. (R)-pyrrolidine-3-carboxylic acid methylamide hydrochloride.

Stage 1 - amide coupling. (R)-Pyrrolidine-1,3-dicarboxylic acid 1-tert-butyl ester (800 mg, 3.61 mmol) was dissolved in THF (5.00 mL) and TEA (4.05 mL, 28.84 mmol) and a solution of methylamine in THF (2 M) was added ; 3.61 ml; 7.21 mmol). A solution of 1-propanephosphonic acid cyclic anhydride (50% in THF; 4.21 mL; 7.21 mmol) was added to the reaction mixture at RT. The reaction mixture was stirred at RT for 1 hour and diluted with 4 N aq. sodium hydroxide (20 ml). Vodn. the phase was extracted with MTBE (2x20 ml) and the combined organic phases were washed with brine, dried, filtered and concentrated in vacuo.

Stage 2 - removal of BOS protection. The crude material from Step 1 was diluted with EtOAc (20 mL) and treated with 4 N HCl in 1,4-dioxane (2 mL; 8.00 mmol) at RT. The reaction mixture was stirred at RT overnight. The reaction mixture was concentrated in vacuo to give intermediate I.10.

Yield: 755 mg (99%), ESI-MS: m/z = 129 [M+H]+, By (HPLC): 0.12 min (HPLC-6).

Intermediate I.11. Morpholin-4-yl-(S)-pyrrolidin-3-yl-methanone hydrochloride.

amide combination. To a solution of (S)-pyrrolidine-1,3 Step 1 dicarboxylic acid 1-tert-butyl ester (500 mg; 2.32 mmol) and TEA (2.61 ml; 18.58 mmol) in THF (4.5 ml) was added a solution of morpholine (220 mg ; 2.56 mmol) in THF (0.8 ml) and then 1-propanephosphonic acid cyclic anhydride (50% in THF; 2.71 ml; 4.65 mmol) was added. The reaction mixture was stirred at RT for 3 hours, diluted with aq. 4 N NaOH (20 ml) and extracted with MTBE (2x20 ml). The combined organic phases were washed with brine, dried with Na ₂ SO ₄ , filtered and evaporated to dryness.

Stage 2 - removal of BOS protection. The crude material from step 1 was resuspended in MeOH (20 mL) and hydrogen chloride (4 N in 1,4-dioxane; 5 mL; 20.00 mmol) was added. The reaction mixture was stirred at RT overnight, concentrated under reduced pressure and co-evaporated with toluene to give intermediate I.11.

Yield: 527 mg (82%), ESI-MS: m/z = 185 [M+H] ⁺ , By (HPLC): 0.12 min (HPLC-6).

The following intermediate was prepared similarly to the procedure described above using the appropriate starting materials. Commentary on the synthesis about changes in this procedure.

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Intermediate connection Structure Starting materials V _y [min] (HPLC method) ms Commentary on the synthesis 1.12 Chiral O ⁿ _X C1 n _o Chiral 2 M dimethylamine THF 0.12 (HPLC-6) 143 Stage 1: 2 eq. amine 1.13 Chiral He ^С1 n o Chiral BUT-O ABOUT 2 M methylamine THF 0.12 (HPLC-6) 129 Stage 1: 2 eq. amine

Intermediate I.14. Racemic cis-3-aza-bicyclo[3.1.0]hex-1-ylmorpholin-4-yl-methanone hydrochloride. Amide combination.

1.14a

Racemic cis-3-aza-bicyclo[3.1.0]hexane-1,3-dicarboxylic acid 3-tert-butyl ester (1.00 g, 4.40 mmol) and HATU (1.90 g, 4.84 mmol) were suspended in DMF and DIPEA was added ( 1.89 ml; 11.00 mmol). The reaction mixture was stirred at RT for 30 min. Morpholine (0.77 mL; 8.80 mmol) was added to the reaction mixture and the solution was stirred at RT overnight. The reaction mixture was diluted with water (20 ml) and extracted with DCM (3x20 ml). The combined organic phases were washed with aq. 1 N NaOH (20 ml), dried and concentrated in vacuo. The crude material was purified by RP-HPLC (C18, 50°C, acetonitrile+0.1% TFA in water) to give intermediate E14a.

Yield: 1.17 g (90%), ESI-MS: m/z = 241 [M+H] ⁺ , By (HPLC): 0.90 min (HPLC-6).

Removing BOS protection.

1.14a 1.14

Intermediate I.14a (0.76 g; 2.56 mmol) was dissolved in MeOH (2.00 mL) and hydrogen chloride (4 N in 1,4-dioxane; 5.00 mL; 20.00 mmol) was added. The reaction mixture was stirred at RT overnight. The reaction mixture was diluted with MTBE, the precipitate was filtered and washed with MTBE. The solvent was allowed to evaporate to obtain intermediate I.14 as a dry solid.

Yield: 0.53 g (89%), ESI-MS: m/z = 197 [M+H] ⁺ , Wu (HPLC): 0.20 min (HPLC-1).

I. 15. (4-Azetidin-3 -yl-piperidin-1 -yl)cyclopropyl-methanone.

Acylation.

3-Piperidin-4-yl-azetidin-1-carboxylic acid tert-butyl ester (530 mg; 2.21 mmol) was dissolved in DCM (20 mL) and TEA (0.71 mL; 5.07 mmol) was added. The solution was cooled with an ice bath and cyclopropanecarbonyl chloride (300 mg; 2.87 mmol) dissolved in DCM (1 ml) was added. The reaction mixture was stirred at 0°C for 1 hour and at 15°C stirred for 3 days. The reaction mixture was diluted with DCM and washed once with us. aq. NaHCO ₃ solution, twice with aq. 0.5 N HCl solution and once with brine. The organic phase was dried over Na ₂ SO ₄ and concentrated in vacuo to give intermediate E15a.

Yield: 690 mg (91%), ESI-MS: m/z = 309 [M+H] ⁺ , By (HPLC): 0.64 min (HPLC-2).

Removing BOS protection.

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Intermediate I.15a (690 mg; 3.01 mmol), TFA (0.62 mL; 8.05 mmol) and DCM (20 mL) were stirred at RT overnight and evaporated to give intermediate I.15.

Yield: 500 mg (77%), ESI-MS: m/z = 209 [M+H]+, By (HPLC): 0.26 min (HPLC-2).

Intermediate I.16. (R)-pyrrolidine-3-carboxylic acid amide trifluoroacetate.

Stage 1 - amide coupling. (R)-Pyrrolidine-1,3-dicarboxylic acid 1-tert-butyl ester (800 mg; 3.61 mmol) was diluted with DCM (8 mL) and N-methylmorpholine (0.45 mL; 3.97 mmol) was added. The reaction mixture was cooled to 0°C and IBCF (0.5 ml; 3.79 mmol) was added. The reaction mixture was stirred at 0°C for 5 min, heated to RT and stirred for 1 hour at RT. After adding aq. NH4OH (32%; 0.67 ml; 5.41 mmol), the reaction mixture was stirred at RT for 80 min. The reaction mixture was diluted with water and extracted with DCM (2x20 ml). The combined organic phases were washed with us. aq. NaHCO ₃ solution, dried and evaporated under reduced pressure.

Stage 2 - removal of BOS protection. The crude material from step 1 was dissolved in DCM (5 mL), TFA (0.83 mL; 10.81 mmol) was added, and the reaction mixture was stirred at RT for 1 h. TFA (0.83 mL; 10.81 mmol) was added to the reaction mixture and stirred at CT scan overnight. The reaction mixture was concentrated in vacuo to give intermediate I.16.

Yield: 1.19 g (100%), ESI-MS: m/z = 115 [M+H]+, By (HPLC): 0.11 min (HPLC-6).

1. 17. Morpholin-4-yl-(8)-pyrrolidin-3-yl-methanone.

Stage 1 - amide coupling. 4-Methyl-piperidine-1,4-dicarboxylic acid mono-tert-butyl ester (500 mg, 1.99 mmol) was dissolved in THF (5 ml) and TEA (2.24 ml, 15.95 mmol), and a solution of ethylamine in THF (2 M; 1.99 ml; 3.99 mmol). A solution of 1-propanephosphonic acid cyclic anhydride (50% in THF; 2.33 mL; 3.99 mmol) was added to the reaction mixture at RT. The reaction mixture was stirred at RT for 1 hour and diluted with 4 N aq. sodium hydroxide (20 ml). Vodn. the phase was extracted with MTBE (2x20 ml) and the combined organic phases were washed with brine (20 ml), dried, filtered and concentrated in vacuo.

Stage 2 - removal of BOS protection. The crude from Step 1 was diluted with EtOAc (20 mL) and treated with 4 N HCl in 1,4-dioxane (1 mL; 4.00 mmol) at RT. The reaction mixture was stirred at RT overnight. The reaction mixture was concentrated in vacuo to give intermediate I.17.

Yield: 236 mg (46%), ESI-MS: m/z = 171 [M+H]+, By (HPLC): 0.13 min (HPLC-6).

Intermediate II.1. Trans-3-(6-chloro-pyridine-3-sulfonyl-3-aza-bicyclo[3.1.0]hexane-6-carboxylic acid methylamide

A mixture of intermediate I.1 (550 mg; 2.49 mmol) and TEA (1.91 ml; 13.58 mmol) in DCM (15 ml) was cooled to 0-5°C. 6-chloropyridine-3-sulfonyl chloride (500 mg; 2.26 mmol) was added to the reaction mixture and stirred for 10 min at 0°C, then warmed to RT and stirred at RT overnight. The reaction mixture was diluted with DCM and washed with water and 1 N aq. HCl. The organic phase was dried with Na ₂ SO ₄ , filtered and reduced in vacuo. The crude material was triturated with diisopropyl ether, the solid was filtered, washed with diisopropyl ether and dried at 60 °C in vacuo to give intermediate II.1.

Yield: 507 mg (71%), ESI-MS: m/z = 316 [M+H]+, By (HPLC): 0.83 min (HPLC-6).

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The following intermediates were prepared similarly to the procedure described above using 6-chloropyridine-3-sulfonyl chloride and the appropriate starting material. For changes to this procedure, see the synthesis commentary.

Intermediate connection Structure Starting material V _y [min] (HPLC method) MS Commentary on the synthesis P.2 n o ^{x x} o About\/ΝΗ 3 eq. TEA; NMP; CT; 2 hours; used as such in the next stage P.Z A /1 \ XL ζξε <J.....\( ζ ^ω o Q ΞΕ 1.2 0.47 (HPLC-1) 315 1 hour; processing: water extraction; crude substance triturated with diisopropyl ether P.4 ,d ^A \\/ ω % n Yu Ν —ά V ^N \Z\ \/ ^ΝΗ 3 eq. TEA; NMP; 1 hour; used as such in next stage P.5 % °Έ° ^Н ')__ JL J AS “ О О L3 0.84 (HPLC-6) 362 3 eq. TEA; CT; 30 min; used as such in the next stage P.6 O Chiral cur O ^{z X} O I.4 3 eq. TEA; NMP; CT; 2 hours; used as such in the next stage P.7 n___,CI ΙΪ | 1 [| J O < A J Η Η ° dx ¹ · 3 eq. TEA; NMP; CT; 2 hours; used as such in the next stage P.8 1 П I l if Ύ o % F ddX O 1 ^H2N yp ^N ^X% ° d^ ^N| - 0.83 (HPLC-6) 333 2 eq. TEA; 1 hour; processing: neutral extraction; drying at 50°C P.9 /— ^zx ° \=/ \=o 0 1.5 1.06 (HPLC-6) 450 3 eq. TEA; II. 10 A d О 2—/ ω d ο d^ ^N ^[^d O^J dx ^h HCl 0.87 (HPLC-6) 374 II. eleven h ω /\\ d ZI o n 1 ^ΝΗ ns 0.88 (HPLC-6) 332 P.12 d o 2—/ ω d o HCl 0.88 (HPLC-6) 332

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11.13 Η 1.6 3 eq. TEA; NMP; KT; 2 hours; used as such in the next stage 11.14 0 ,ν. yu ο^ο 1.7 1.5 eq. TEA; processing: water, extraction 11.15 ο.Ν. .CI Η 1 to /Ν. ο Ο -'''^ ^Η ΗΒ 3 eq. TEA; NMP; CT; 2 hours; used as such in the next stage 11.16 ω f —ζ \ 1.8 3 eq. TEA; NMR; 1 hour; used as such in the next stage 11.17 ο 4 ζ, ΐ ___^ΝΗ 0.79 (HPLC-6) 304 3 eq. TEA; 1 hour 20 minutes 11.18 A 0 \\ / ω t> ο V-NH HCl 3 eq. TEA; NMP; CT; 2 hours; used as such in the next stage Π. 19 ο A /L e τζ ) 1.9 3 eq. TEA; NMP; CT; 2 hours; used as such in the next stage

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11.20 1 Y 1 i if η 1 ύΥί Ο \z ^NH 3 eq. TEA; NMP; KT; 2 hours; used as such in the next stage 11.21 .n. .ci K Y 1 [f V ° k AN Λ A O·' o —a 1, Y^ o V^H HCl 3 eq. TEA; NMP; 1 hour; used as such in the next stage 11.22 ^H o a °\ z/ ^X cc A xn 0\/NH 0.60 (HPLC-1) 332 2 eq. TEA; CT; 1.5 h treatment: aq. acid extraction 11.23 iz^o ,0 w/ ω Aо ^Y trVNH 1.5 eq. TEA; NMP; 1 hour; used as such in the next stage 11.24 Chiral _.N. .Cl Y<1,LT Y °* *0 LIO 0.81 (HPLC-6) 304 4.00 eq. TEA; CT 40 min; used as such in the next stage 11.25 /— _o Chiral ° N-/ / 5__ .N. .Cl α,χχ CY o Lil 0.87 (HPLC-6) 360 3.00 eq. TEA; THF/DMSO; 440 min; used as such in the next stage 11.26 IZ .N^.CI hctY-T [f η Vik X hoYt VnH HCl 3 eq. TEA; NMP; CT; 2 hours; used as such in the next stage

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11.27 \o Chiral N—D <1»У 0^ 0 1.12 0.87 (HPLC-6) 318 DMSO; 40 min; used as such in the next stage 11.28 / -Z/° o z-^ I % o X I %ΝΗ _Х С1 n 0.84 (HPLC-6) 304 THF; 37 min; used as such in the next stage 11.29 ιζ ^χ сА °\\/ ^z ω th ο Chiral 1.13 0.81 (HPLC-6) 304 THF/DMSO; 40 min; used as such in the next stage 11.30 0 V=o , /' ,N. ,CI ^Η 0 ^x 0 1.14 0.48 (HPLC-1) 371 CT; processing: neutral aq. extraction; HPLC purification (ACN/water + TFA) 11.31 kF b O Z^ ω % o 0 ^H 2 ^N ^\% Vnh „a Η 0.73 (HPLC-6) 276 2.10 eq. TEA; THF; CT; 36 min; used as such in the next stage 11.32 A 0^0 1.15 3 eq. TEA; NMP; CT; 2 hours; used as such in the next stage 11.33 O Chiral ^H 2 ^N ^ \ ,C1 ai o' o 1.16 0.77 (HPLC-6) 290 5 eq. TEA; processing: neutral aq. extraction; the precipitate is filtered, evaporated together with iPrOH, Tol

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11.34 /IN V% 1.17 0.93 (HPLC-6) 346 4 eq. TEA; DMSO/THF; KT; 25 min; used as such in the next stage 11.35 BUT Chiral α,ι> ^h o BUT Chiral He 0.78 (HPLC-6) 263 2 eq. TEA; 1.5 h 11.36 BUT Chiral \/N. ,C1 Cdh 0 ^{x X} O |—IQ Chiral ^nh 0.78 (HPLC-6) 263 11.37 A \\ / ω o fVnh HCl 0.51 (HPLC-1) 269 CT; 1 hour; processing: neutral aq. extraction; crude substance triturated with diisopropyl ether 11.38 o 0 \\ / ω oh oh Ojh 0.43 (HPLC-1) 233 2 eq. TEA; CT; 1 hour; processing: neutral aq. extraction; crude substance triturated with diisopropyl ether 11.39 -O \ ,Ν. ,С1 U./J ss —o ^Inh HCl 0.47 (HPLC-1) 277 2 eq. TEA; CT; 1 hour; treatment: neutral aq. extraction 11.40 ,___ SAME ,C1 SG ^{h o} Xi HCl CT; 1 h, treatment: neutral. aq. extraction; crude substance

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ground with diisopropyl ether 11.41 O % ω /\\ 7° O \ V-NH NS 0.44 (HPLC-1) 263 2 eq. TEA; CT 1 hour; processing: water, neutral. extraction; crude substance triturated with diisopropyl ether 11.42 O \\/ ω °0 o V-NH HCI 0.53 (HPLC-1) 283 3 eq. TEA; CT; 1 hour; processing: water. neutral extraction; crude substance triturated with diisopropyl ether 11.43 O h % % 0.82 (HPLC-6) 318 3 eq. TEA 11.44 O th ω σ o I ^but \/0 \/ ^NH 0.81 (HPLC-6) 277 3 eq. TEA 11.45 \ o ,0 ω ”0 O \/ ^ΝΗ 0.53 (HPLC-1) 291 2 eq. TEA; CT; 1 hour; processing: neutral aq. extraction with DCM; 11.46 .O \\/ ω % o 0.58 (HPLC-1) 261 2 eq. TEA; CT; 1 hour; treatment: rinsing with water; crude substance ground with diisopropyl ether 11.47 O 0. ω /\\ 0 ΞΕ ^η °Τ2?νη 3 eq. TEA 11.48 ,O Chiral η ₂ ν0 'h DK / ^C| s/L 0 ^x '0 Ο Η ₂ Ν-0 b~ HCI 0.43 (HPLC-2) 290 3 eq. TEA 11.49 <0 p 0 O HCI 0.63 (HPLC-2) 345 3 eq. TEA

Intermediate III.1. 1-(6-fluoro-pyridine-3-sulfonyl)piperidine-4carboxylic acid methyl ester.

111.1

Piperidine-4-carboxylic acid methyl ester hydrochloride (1.15 g; 6.39 mmol) was suspended in DCM (40 ml) and TEA (3.56 ml; 25.56 mmol) was added. A solution of 6fluoropyridine-3-sulfonyl chloride (1.25 g, 6.39 mmol) in DCM (10 ml) was added to the reaction mixture. It was stirred at RT for 45 min, then diluted with DCM (50 ml) and washed with water (2x40 ml). The combined organic phases were dried with Na ₂ SO ₄ and concentrated in vacuo. The crude material was suspended in MTBE and the remaining solid was filtered to obtain intermediate III.1.

Yield: 1.4 g (73%), ESI-MS: m/z = 302 [M+H]+, By (HPLC): 0.52 min (HPLC-1).

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The following intermediates were prepared similarly to the procedure described above using 6-fluoropyridine-3-sulfonyl chloride and the appropriate starting material. For changes to this procedure, see the synthesis commentary.

Intermediate connection Structure Starting material B _y [min] (HPLC method) ms Commentary on the synthesis III. 2 O 11 ⁿ HA XJ +1. Η^ΝΗ HCl 0.53 (HPLC-1) 301 2 eq. TEA; CT; 1.5 h Sh.Z A ω У O / ABOUT 0.53 (HPLC-1) 301 2 eq. TEA; CT; 2.5 h; III. 4 \ o ο _χ W “P HCl 0.67 (HPLC-1) 317 2 eq. TEA; 3.5 h

Intermediate IV.1. Tert-butyl-X-[2-(fluoromethylidene)-3-hydroxypropyl]carbamate (E/Z mixture).

An E/Z alcohol mixture (intermediate IV.1) was prepared according to the procedure described in WO 2013/163675, cc. 50-53.

Allyl)carbamic acid intermediate.

IV.2. Tert-butyl ether ((E)-3-fluoro-2-hydroxymethyl-

Intermediate IV.1 (4.00 g; 19.49 mmol) was purified three times by silica gel column chromatography to give the single E-isomer IV.2 (1.95 g; 9.50 mmol; 49%).

Example 1. Trans-3-[6-((E)-2-aminomethyl-3-fluoroallyloxy)pyridin-3sulfonyl]-3-aza-bicyclo[3.1.0]hexane-6-carboxylic acid methyl amide trifluoroacetate.

Substitution.

Intermediate II.1 (326 mg; 85% purity; 0.88 mmol) and intermediate IV.1 (216 mg; 1.05 mmol) were dissolved in THF (1 ml; S) and DMSO (1 ml; S) and cooled up to 0°C. Sodium tert-butoxide (2M V THF; 0.53 mL; 1.05 mmol; V) was added to the reaction mixture and after 5 min at 0°C the mixture was stirred at RT for 35 min (t). The reaction mixture was purified by HPLC (ACN/water+TFA) to obtain BOC-protected Example 1.

Yield: 410 mg (96%), ESI-MS: m/z = 385 [M+H-BOC] ⁺ , Wu (HPLC): 1.05 min (HPLC-6).

Removing BOS protection.

The BOC-protected compound of Example 1 as an E/Z mixture (410 mg; 0.85 mmol) was dissolved in DCM (15 mL; S) and TFA (266 μL; 3.45 mmol; A) was added. The reaction mixture was stirred at RT for 2.5 hours (t), then evaporated under reduced pressure, dissolved in MeOH (5 ml) and purified by RP-HPLC (ACN/water+TFA) to give Example 1.

Yield: 160 mg (38%), ESI-MS: m/z = 385 [M+H] ⁺ , By (HPLC): 0.64 min (HPLC-5).

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The following examples (the example number is indicated in column No.) were prepared similarly to the procedure described above using the appropriate starting materials. Details of the two steps are given in the column Synthesis Comment, Retention Time and Mass (ESI-MS, m/z =

[M+H]+) determined by HPLC-MS are indicated in the CT and MS columns.

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No. Substitution Removing BOS protection Starting materials V _y [min] (HPLC method) MS Commentary on the synthesis V _y [min] (HPLC method) MS Commentary on the synthesis 2 II.2; IV.1 0.77 (HPLC8) 501 S: THF/ NMP B: 4.10 eq. T: 0°C to RT 1: overnight cleanup: HPLC (ACN/water + NH ₄ OH) 0.40 (HPLC-7) 401 S: DXM A: 51 eq. TFU T: CT 1: 1 h 3 II.3; IV.1 0.61 (HPLC1) 485 S: THF/ NMP B: 4.00 eq. T: CT 1: 30 min processing: evaporation; without cleaning 0.37 (HPLC-1) 385 S: DXM A: 13 eq. TFU T: CT 1: 1 h 4 IL4; IV.1 0.96 (HPLC4) 414 S: THF/ NMP B: 4.00 eq. T: from 0°C to RT t: 2 h 0.70 (HPLC-6) 414 S: DXM A: 23 eq. TFU T: CT t: 2 h 5 II.5; IV.1 1.01 (HPLC- 6) 531 S: DXM B: 4.00 eq. T: CT 1:45 min 0.69 (HPLC-6) 431 S: DXM A: 6 eq. TFU T: CT 1:2 h 6 II.6; IV.1 0.74 (HPLC8) 487 S: THF/ NMP B: 4.10 eq. T: 0°C to RT 1: overnight purification: HPLC (ACN/water + NH ₄ OH) 0.36 (HPLC-7) 387 S: DXM A: 51 eq. TFU T: CT 1: 1 h 7 IL7; IV.1 0.71 (HPLC8) 502 S: THF/ NMP B: 4.10 eq. T: 0°C to RT 1: overnight purification: HPLC (ACN/water + NH ₄ OH) 0.36 (HPLC-7) 402 S: DXM A: 51 eq. TFU T: CT 1: 1 h

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8 II.8; IV.l 0.94 (HPLC4) 402 S: THF/ NMP B: 4.00 eq. T: 0°C to KT t: 2 h 0.68 (HPLC-6) 402 S: DXM A: 14 eq. TFU T: CT 1:2 h 9 II.9; IV.l 1.15 (HPLC- 6) 519 S: THF/DMSO B: 1.05 eq. T: 0°C to KT t: 80 min h 0.84 (HPLC-6) 519 S: DXM A: 7 eq. TFU T: CT 1:2 h 10 II.10; IV.l 1.07 (HPLC6) 443 S: THF/DMSO B: 1.05 eq. T: 0°C to KT t: 35 min h 0.72 (HPLC-6) 443 S: DXM A: 26 eq. TFU T: CT eleven II.11; IV.l 1.07 (HPLC6) 401 S: THF/DMSO B: 1.05 eq. T: from 0°C to CT 1: 40 min h 0.71 (HPLC-6) 401 S: DXM A: 8 eq. TFU T: CT 12 11.12; IV.l 1.08 (HPLC- 6) 401 S: THF/DMSO B: 1.05 eq. T: from 0°C to CT 1:40 min 0.71 (HPLC-6) 401 S: DXM A: 25 eq. TFU T: CT 13 11.13; IV.l 0.77 (HPLC8) 513 S: THF/ NMP B: 4.10 eq. T: 0°C to RT 1: overnight purification: HPLC (ACN/water + NH ₄ OH) 0.40 (HPLC-7) 413 S: DXM A: 51 eq. TFU T: CT 1: 1 h

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14 11.14; IV.l S: DXM B: 6.00 eq. T: 0°C to KT t: 2 d treatment: aq. extraction; without cleaning 0.38 (HPLC-2) 427 S: DXM A: 5 eq. TFU T: CT 1: during. nights 15 11.15; IV.l 0.77 (HPLC8) 513 S: THF/ NMP B: 4.10 eq. T: 0°C to RT 1: overnight cleanup: HPLC (ACN/water + NH ₄ OH) 0.37 (HPLC-7) 401 S: DXM A: 51 eq. TFU T: CT 1: 1 h 16 11.16; IV.l 1.05 (HPLC- 6) 515 S: THF/ NMP B: 4.00 eq. T: from 0°C to RT 1:2 h 0.74 (HPLC-6) 415 S: DXM A: 44 eq. TFU T: CT 1:2 h 17 11.17; IV.l 0.97 (HPLC6) 373 S: THF/DMSO B: 1.05 eq. T: from 0°C to CT 1:35 min 0.66 (HPLC-6) 373 S: DXM A: 19 eq. TFU T: CT 1: 1 h 18 11.18; IV.l 0.74 (HPLC8) 432 S: THF/ NMP B: 4.10 eq. T: 0°C to RT 1: overnight cleanup: HPLC (ACN/water + NH ₄ OH) 0.34 (HPLC-7) 332 S: DXM A: 51 eq. TFU T: CT 1: 1 h 19 11.19; IV.l 0.78 (HPLC8) 515 S: THF/ NMP B: 4.10 eq. T: 0°C to RT 1: overnight cleanup: HPLC (ACN/water + NH ₄ OH) 0.41 (HPLC-7) 415 S: DXM A: 51 eq. TFU T: CT 1: 1 h

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20 11.20; IV.l 0.76 (HPLC8) 501 S: THF/ NMP B: 4.10 eq. T: 0°C to KT t: overnight purification: HPLC (ACN/water + NH ₄ OH) 0.40 (HPLC-7) 401 S: DXM A: 51 eq. TFU T: CT 1: 1 h 21 11.21; IV.l 1.05 (HPLC- 6) 528 S: THF/ NMP B: 4.00 eq. T: from 0°C to rt 1:2 h 0.73 (HPLC-6) 428 S: DXM A: 44 eq. TFU T: CT 1: 1 h 22 11.22; IV.l 0.71 (HPLC1) 502 S: Tol B: 2.50 eq. in the form of a solid T: from 0°C to rt 1: 3 h treatment: aq. acid extraction; HPLC purification (ACN/water + TFA) 0.44 (HPLC-1) 402 S: 1.4dioxane A: 60 eq. NS1 T: CT 1: 1 h 23 11.23; IV.l 0.98 (HPLC6) 473 S: THF/ NMP B: 3.00 eq. T: from 0°C to ct 1: overnight 0.64 (HPLC-6) 373 S: DXM A: 44 eq. TFU T: CT 1: 1 h 24 11.24; IV.l 1.00 (HPLC- 6) 473 S: DXM B: 4.00 eq. T: CT 1: overnight 0.65 (HPLC-6) 373 S: DXM A: 30 eq. TFU T: CT 1: 1 h

- 50 043735

25 11.25; IV.1 1.04 (HPLC- 6) 529 S: THF/DMSO B: 4.10 eq. T: from 0°C to rt 1: 1.5 h 0.70 (HPLC-6) 429 S: DXM A: 7 eq. TFU T: CT 26 11.26; IV.1 0.87 (HPLC8) 460 S: THF/ NMP B: 4.10 eq. T: 0°C to RT 1: overnight purification: HPLC (ACN/water + NH ₄ OH) 0.38 (HPLC-7) 360 S: DXM A: 51 eq. TFU T: CT t: 1 h 27 11.27; IV.1 1.04 (HPLC- 6) 487 S: THF/DMSO B: 4.10 eq. T: from 0°C to CT 1: 1.5 h 0.70 (HPLC-6) 387 S: DXM A: 9 eq. TFU T: CT t: 75 min 28 11.28; IV.1 1.03 (HPLC- 6) 473 S: THF/DMSO B: 4.10 eq. T: from 0°C to CT 1: 1.5 h 0.67 (HPLC-6) 373 S: DXM A: 9 eq. TFU T: CT 29 11.29; IV.1 1.01 (HPLC- 6) 473 S: THF/DMSO B: 4.10 eq. T: from 0°C to CT 1: 1.5 h 0.66 (HPLC-6) 373 S: DXM A: 11 eq. TFU T: CT thirty 11.30; IV.1 0.63 (HPLC1) 541 S: THF B: 4.00 eq. T: CT 1:40 min 0.43 (HPLC-1) 441 S: DXM A: 13 eq. TFU T: CT 1:2 h 31 11.31; IV.1 0.97 (HPLC6) 445 S: THF/DMSO B: 4.10 eq. T: from 0°C to CT 1: 2.5 h 0.62 (HPLC-6) 345 S: DXM A: 60 eq. TFU T: CT 32 11.32; IV.1 0.84 (HPLC8) 553 S: THF/ NMP B: 4.10 eq. T: 0°C to RT 1: overnight purification: HPLC (ACN/water + NH ₄ OH) 0.49 (HPLC-7) 453 S: DXM A: 51 eq. TFU T: CT t: 1 h 33 11.33; IV.1 0.96 (HPLC6) 459 S: DMSO B: 1.05 eq. T: CT 1: overnight 0.63 (HPLC-6) 359 S: DXM A: 18 eq. TFU T: CT 1:2 h 34 11.34; IV.1 1.07 (HPLC6) 515 S: THF/ DMSO/ DCM B: 4.00 eq. T: CT 1: overnight 0.74 (HPLC-6) 415 S: DXM A: 5 eq. TFU T: CT 1: 2 h

Example 35. (S)-1-[6-((E)-2-aminomethyl-3-fluoroallyloxy)pyridin-3-sulfonyl]pyrrolidin-3-ol. Substitution.

- 51 043735

Intermediate IV.2 (176 mg; 0.86 mmol) was diluted with THF (6 mL; S) and sodium hydride (55%; 75 mg; 1.72 mmol; B) was added at RT. After stirring at RT for 10 min (t), intermediate II.35 (226 mg; 0.86 mmol) was added. The reaction mixture was stirred at RT overnight (t) and purified by RP-HPLC (ACN/water+TFA) to give BOC protected Example 35.

Yield: 139 mg (38%), ESI-MS: m/z = 432 [M+H] ⁺ , By (HPLC): 0.63 min (HPLC-2).

Removing BOS protection.

The BOC-protected compound of Example 35 (139 mg; 0.32 mmol) was diluted with DCM (4 ml; S) and TFA (1.5 ml; 195 mmol; A) was added. The reaction mixture was stirred at RT for 3 hours and purified by RP-HPLC (ACN/water+TFA) to give Example 35.

Yield: 103 mg (27%), ESI-MS: m/z = 332 [M+H] ⁺ , By (HPLC): 0.62 min (HPLC-6).

The following examples (the example number is indicated in column No.) were prepared similarly to the procedure described above using the appropriate starting materials. Details of the two steps are given in the synthesis comment column, retention time and mass (ESI-MS, m/z = [M+H]+) determined by HPLC-MS are given in the By and MS columns.

- 52 043735

- 53 043735

No. Substitution Removing BOS protection Starting materials By [min] (HPLC method) MS Commentary on the synthesis By [min] (HPLC method) ms Commentary on the synthesis 36 11.36; IV.2 0.63 (HPLC-2) 432 S: THF B: 2.00 eq. T: CT 1: during nights 0.62 (HPLC-6) 332 S: DXM A: 9 eq. TFU T: CT 1: 3 h 37 11.37; IV.2 S: DMF B: 1.00 eq. T: CT t: 2 h; intermediate connection is not allocated 0.34 (HPLC-1) 338 S: DXM A: 42 eq. TFU T: CT t: 2 h 38 O. XX IV.2 S: DMF B: 1.00 eq. T: CT t: 2 h; intermediate connection is not allocated 0.34 (HPLC-3) 316 S: DXM A: 42 eq. TFU T: CT t: 2 h 39 11.38; IV.2 S: DMF B: 2.00 eq. T: CT t: 2 h; intermediate connection is not allocated 0.39 (HPLC-1) 302 S: DXM A: 37 eq. TFU T: CT t: 1 h 40 11.39; IV.2 S: DMF B: 2.00 eq. T: CT t: 2 h; intermediate connection is not allocated 0.33 (HPLC-1) 346 S: DXM A: 43 eq. TFU T: CT t: 2 h 41 11.40; IV.2 S: DMF B: 2.00 eq. T: CT t: 2 h; intermediate connection is not allocated 0.38 (HPLC-1) 328 S: DXM A: 2 eq. TFU T: CT t: 2 h 42 11.41; IV.2 S: DMF B: 2.00 eq. T: CT t: 2 h; intermediate connection is not allocated 0.36 (HPLC-1) 332 S: DXM A: 41 eq. TFU T: CT t: 2 h 43 11.42; IV.2 S: DMF B: 2.00 eq. T: CT t: 2 h; intermediate connection is not allocated 0.40 (HPLC-1) 352 S: DXM A: 44 eq. TFU T: CT t: 1 h

Example 44. 1-[6-((E)-2-aminomethyl-3-fluoroallyloxy)pyridine-3sulfonyl]piperidine-4-carboxylic acid methyl amide trifluoroacetate.

Substitution.

11-43 IV.1 BOC-protected F example 44

Intermediate IV.1 (70 mg; 0.34 mmol) was dissolved in THF (1 ml; S) and sodium hydride (55%; 30 mg; 0.68 mmol; B) was added. After stirring at RT (T) for 10 min (T), intermediate II.43 (108 mg, 0.34 mmol) was added and the reaction mixture was stirred at RT (T) overnight (T). The reaction mixture was purified by RP-HPLC (ACN/water+TFA) to give BOC-protected Example 44.

Yield: 95 mg (57%), ESI-MS: m/z = 487 [M+H] ⁺ , By (HPLC): 0.64 min (HPLC-2).

Removing BOS protection.

- 54 043735

The BOC-protected compound of Example 44 as an E/Z mixture (95 mg; 0.20 mmol) was diluted with DCM (4 mL; S) and TFA (1.5 mL; 19.47 mmol; A) was added. The reaction mixture was stirred at RT for 3.3 hours and purified by RP-HPLC (ACN/water+TFA) to give Example 44.

Yield: 44 mg (26%), ESI-MS: m/z = 387 [M+H]+, By (HPLC): 0.66 min (HPLC-6).

The following examples (the example number is indicated in column No.) were prepared similarly to the procedure described above using the appropriate starting materials. Details of the two steps are given in the synthesis comment column, retention time and mass (ESI-MS, m/z = [M+H]+) determined by HPLC-MS are given in the By and MS columns.

- 55 043735

No. Substitution Removing BOS protection Starting materials By [min] (HPLC method) MS Commentary on the synthesis By [min] (HPLC method) MS Commentary on the synthesis 45 11.44; VI.1 0.65 (HPLC-2) 446 S: THF B: 2.00 eq. T: CT 1: overnight 0.65 (HPLC-6) 346 S: DXM A: 26 eq. TFU T: CT t: overnight 46 11.45; IV.1 S: THF B: 2.00 eq. T: CT 1: 2 h; intermediate connection is not allocated 0.37 (HPLC-1) 360 S: DXM A: 46 eq. TFU T: CT 1:2 h 47 11.46; IV.1 S: THF B: 2.00 eq. T: CT 1: 2 h; intermediate connection is not allocated 0.40 (HPLC-1) 330 S: DXM A: 41 eq. TFU T: CT 1:2 h 48 11.47; IV.1 0.68 (HPLC-2) 460 S: THF B: 2.00 eq. T: CT 1: overnight 0.70 (HPLC-6) 360 S: DXM A: 19 eq. TFU T: CT t: 1 h 49 11.48; IV.1 0.61 (HPLC-2) 459 S: THF B: 2.00 eq. T: CT 1: overnight 0.62 (HPLC-6) 359 S: DXM A: 35 eq. TFU T: CT t: 1 h 50 11.49; IV.1 0.75 (HPLC-2) 514 S: THF B: 2.00 eq. T: CT 1: overnight 0.78 (HPLC-6) 414 S: DXM A: 24 eq. TFU T: CT 1:2 h

Example 51. 1-[6-((E)-2-aminomethyl-3-fluoroallyloxy)pyridine-3sulfonyl]piperidine-4-carboxylic acid methyl ester trifluoroacetate.

Substitution.

Ш · ^{1 ιν} · ¹ BOC-protected example 51

Intermediate IV.1 (70 mg; 0.33 mmol) was dissolved in Tol (3 ml; S) and sodium tert-butoxide (30 mg; 0.33 mmol; B) was added. Intermediate III.1 (100 mg; 0.33 mmol) was added to the reaction mixture and the reaction mixture was stirred at RT for 35 min (t). The toluene was evaporated under reduced pressure, the residue was resuspended in MeOH (3 ml) and purified by RPHPLC (ACN/water+TFA) to give BOC-protected Example 51.

Yield: 97 mg (60%), ESI-MS: m/z = 488 [M+H] ⁺ , By (HPLC): 0.69 min (HPLC-1).

Removing BOS protection.

BOC-protected example 51 example 51

The BOC-protected compound of Example 51 (50 mg; 0.10 mmol) was diluted with DCM (4 ml; S) and TFA (30 μl; 0.41 mmol; A) was added. The reaction mixture was stirred at RT (T) over the weekend (t). It was then evaporated in vacuo and the residue was resuspended with MeOH (3 ml) and purified by RP-HPLC (ACN/water+TFA) to give Example 51.

Yield: 16 mg (31%), ESI-MS: m/z = 388 [M+H] ⁺ , By (HPLC): 0.4 min (HPLC-1).

The following examples (the example number is indicated in column No.) were prepared similarly to the procedure described above using the appropriate starting materials. Details of the two steps are given in the synthesis comment column, retention time and mass (ESI-MS, m/z = [M+H] ⁺ ) determined by HPLC-MS are indicated in the By and MS columns.

Intermediate V.1. {1-[6-(2-Aminomethyl-3-fluoroallyloxy)pyridin-3-sulfonyl]piperidin-4-yl]acetic acid methyl ester (E/Z mixture).

Alcohol IV.1 (0.95 g; 4.62 mmol) was dissolved in toluene (30 ml) and sodium tert-butoxide (0.44 g; 4.62 mmol) and intermediate III.4 (1.46 g; 4.62 mmol) were added. The reaction mixture was stirred at RT for 2 hours, diluted with toluene (30 ml) and extracted twice with water. The organic phase was dried over Na ₂ SO ₄ and evaporated under reduced pressure. The residue was purified by silica gel chromatography to give intermediate V. 1.

Yield: 1.85 g (80%), ESI-MS: m/z = 502 [M+H] ⁺ , By (HPLC): 0.72 min (HPLC-1).

The following intermediates were prepared similarly to the procedure described above using alcohol IV.1 and the appropriate starting material. For changes to this procedure, see the synthesis commentary.

- 57 043735

Intermediate connection Structure Starting material Wu [min] (HPLC method) ms Commentary on the synthesis V.2 χι. Ό .......G o o 1 Sh.2 0.69 (HPLC-1) 486 1 hour V.3 III. 1 0.71 (HPLC-1) 488 processing: recrystallization with PE/EtOAC (3:1) V.4 °V?, XY sh.z 0.72 (HPLC-1) 502 70 min

Intermediate VI.1. {1-[6-(2-Aminomethyl-3-fluoroallyloxy)pyridin-3sulfonyl]piperidin-4-yl}acetic acid (E/Z mixture).

Intermediate V.1 (1.85 g, 3.69 mmol) was dissolved in MeOH (70 ml) and aq. NaOH (1N; 22.13 ml; 22.13 mmol). The reaction mixture was stirred at RT for 10 min, then acidified with citric acid (10%), and MeOH was evaporated under reduced pressure. The residue was cooled to 5°C, the precipitate was filtered, washed with water (10 ml) and dried at 40°C to obtain intermediate VI.1.

Yield: 1.31 g (73%), ESI-MS: m/z = 488 [M+H]+, By (HPLC): 0.62 min (HPLC-1).

The following intermediates were prepared similarly to the procedure described above using the appropriate starting material. For changes to this procedure, see the synthesis commentary.

Example

Intermediate connection Structure Starting material Wu [min] (HPLC method) ms Commentary on the synthesis VI.2 o' A 7^ V.2 0.62 (HPLC-1) 472 4 eq. NaOH; 18 h VI.3 and [G N ^but T Ί X y Y 7s; ο ^ζ 'o 1 V.3 0.62 (HPLC-1) 474 4 eq. NaOH; 3 hours VI. 4 o' ⁴ o V.4 0.63 (HPLC-1) 487 7.2 eq. NaOH; 2d

54. Trifluoroacetate {1-[6-((E)-2-aminomethyl-3-fluoroallyloxy)piperidin-3sulfonyl] piperidin-4-yl} acetic acid.

Intermediate VI.1 (50 mg; 0.10 mmol) was dissolved in a solution of 4 N hydrogen chloride in 1,4-dioxane (1.5 ml; 6.00 mmol) and stirred at RT for 70 min. The reaction mixture was evaporated in vacuum. The residue was dissolved in MeOH (3 ml) and purified by RP-HPLC (ACN/water+TFA) to give Example 54.

Yield: 18 mg (35%), ESI-MS: m/z = 388 [M+H]+, By (HPLC): 0.40 min (HPLC-1).

Example 55 Trans-3-[6-((E)-2-aminomethyl-3-fluoroallyloxy)pyridin-3-sulfonyl]-3-aza-bicyclo[3.1.0]hexane-6-carboxylic acid trifluoroacetate.

- 58 043735

To a solution of intermediate VI.2 (50 mg; 0.11 mmol) in DCM (10 ml) was added TFA (50 mg; 0.42 mmol). The reaction mixture was stirred at RT for 50 min, evaporated in vacuo and the residue was purified by RP-HPLC (ACN/water+TFA) to give Example 55.

Yield: 14 mg (27%), ESI-MS: m/z = 372 [M+H] ⁺ , By (HPLC): 0.40 min (HPLC-1).

The following examples were prepared similarly to the procedure described above using the appropriate starting material. For changes to this procedure, see the synthesis commentary.

Example Structure Starting material By [min] (HPLC method) MS Commentary on the synthesis 56 -F O < A-O. 41.NH, % o o F VI.3 0.41 (HPLC-1) 374 2 hours 57 O < A.O. A. .ΝΗ, ^but Αλ ХУ o F VI. 4 0.73 (HPLC-5) 388 15.5 hours

Intermediate VII.1. (3-Fluoro-2-{5-[4-methyl-4-(tetrahydropyran-4-ylcarbamoyl)piperidin-1-sulfonyl]pyridin-2-yloxymethyl}allyl)carbamic acid tert-butyl ester (E/Z mixture) .

Intermediate VI.4 (40 mg; 0.08 mmol) was dissolved in DMF (2.00 mL) and TEA (46 μL; 0.33 mmol) and TCFH (23 mg; 0.08 mmol) were added. The reaction mixture was stirred at RT for 10 min, and 4-aminotetrahydropyran (20 mg; 0.20 mmol) was added. The reaction mixture was stirred at RT overnight, then acidified with TFA (aq; 50%) and purified by RP-HPLC (ACN/water+TFA) to give intermediate VII.1.

Yield: 22 mg (47%), ESI-MS: m/z = 471 [M+H] ⁺ , By (HPLC): 1.05 min (HPLC-6).

The following intermediates were prepared similarly to the procedure described above using the appropriate starting materials.

Intermediate connection Structure Starting material By [min] (HPLC method) MS VII.2 ¹ // I/ ^na o A he F 1 VI.2 E05 (HPLC-6) 433

Intermediate VII.3. {2-[5-(4-Carbamoylmethylpiperidin-1-sulfonyl)pyridin-2-yloxymethyl]3-fluoroallyl}carbamic acid tert-butyl ester trifluoroacetate (E/Z mixture).

To a solution of intermediate VI. 1 (100 mg; 0.21 mmol) in DMF (1 ml), TEA (40 μl; 0.41 mmol) and HATU (90 mg; 0.23 mmol) were added at RT. Ammonium (0.5

- 59 043735

M in 1,4-dioxane; 2 ml; 1.00 mmol) and it was stirred at RT for 1 hour 40 minutes. The reaction mixture was diluted with water and extracted with EtOAc. The combined organic phases were dried with Na ₂ SO ₄ and evaporated. The crude material was resuspended in MeOH (3 ml) and purified using

RP-HPLC (ACN/water+TFA) to obtain intermediate VII.3.

Yield: 70 mg (70%), ESI-MS: m/z = 486 [M+H] ⁺ , By (HPLC): 0.58 min (HPLC-1).

The following intermediates were prepared similarly to the procedure described above using the appropriate starting materials. For changes to this procedure, see the synthesis commentary.

Intermediate connection Structure Starting materials By [min] (HPLC method) MS Commentary on the synthesis VII.4 .F U ABOUT ΝΗ ₃ O NH, 0 no Un. VI.4 0.96 (HPLC4) 387 5 eq. amine; 8 eq. TEA; during the night; processing: without extraction purification: HPLC (ACN/water + NH ₄ OH) VII.5 ? if ⁿ \ ⁿ MzZi S ' g VI.3 ' 601 1.5 eq. amine; 4.25 ea TEA; during the night; Purification: HPLC (ACN/water + NH ₄ OH) VII.6 ^th T t VI.3 ' 0.77 (HPLC8) 513 1.5 eq. amine; 4.25 ea TEA; during the night; Purification: HPLC (ACN/water + NH4OH) VII.7 ^F ^nh ₂ HCl. VI.3' 0.77 (HPLC8) 533 1.5 eq. amine; 4.25 ea TEA; during the night; Purification: HPLC (ACN/water + NH4OH) VII. 8 (Π ^f ^nh ₂ F HCl . VI.3' 0.83 (HPLC8) 565 1.5 eq. amine; 4.25 ea TEA; during the night; Purification: HPLC (ACN/water + NH4OH)

Intermediate VII.9. (3-Fluoro-2-{5-[6-(2-methoxyethylcarbamoyl)-3-aza-bicyclo[3.1.0]hexane-3-sulfonyl]pyridin-2-yloxymethyl}allyl)carbamic acid tert-butyl ester ( E/Z mixture).

oh oh

To a solution of intermediate VI.2 (40 mg; 0.08 mmol), 2-methoxyethylamine (15 mg: 0.20 mmol) and N-methylmorpholine (47 μl; 0.42 mmol) in DCM (2 ml) was added 1-propanephosphonic acid cyclic anhydride (50% in EtOAc; 100 µl; 0.17 mmol). The reaction mixture was stirred again at RT overnight, treated with 1-propanephosphonic acid cyclic anhydride (50% in EtOAc; 50 μL; 0.09 mmol) and stirred at RT overnight. The reaction mixture was dissolved in ACN/water and purified by RP-HPLC (ACN/water+NH ₄ OH) to give intermediate VII.9.

ESI-MS: m/z = 552 [M+H]+, By (HPLC): 0.75 min (HPLC-8).

Example 58 1-[6-((E)-2-aminomethyl-3-fluoroallyloxy)pyridin-3-sulfonyl]-4-methyl-piperidine-4-carboxylic acid (tetrahydropyran-4-yl)-amide trifluoroacetate.

νιι.1 example 58

A solution of intermediate VII.1 (22 mg; 0.04 mmol) and TFA (1.00 ml; 12.96 mmol) in DCM (1 ml) was stirred at RT for 2 h, then evaporated to dryness under reduced pressure, acidified with TFA (50 %) and purified by RP-HPLC (ACN/water+TFA) to give Example 58.

Yield: 13 mg (56%), ESI-MS: m/z = 471 [M+H] ⁺ , By (HPLC): 0.74 min (HPLC-6).

The following examples were prepared similarly to the procedure described above using the appropriate starting material. For changes to this procedure, see the synthesis commentary.

Example Structure Starting material By [min] (HPLC method) ms Commentary on the synthesis 59 41. ω g-2 O U Ζ— ABOUT \ VII.2 0.73 (HPLC-6) 443 hut TFU 60 AND VII.3 0.40 (HPLC-1) 386 45 eq. TFU; 100 min 61 .F Η |ί F VII.4 0.68 (HPLC-6) 387 hut TFU 62 ⁰ II D-. /N o^^nh ₂ ^h u _s %i = ° ° F VII.5 0.33 (HPLC-9) 417 77 eq. TFU; 1 hour 63 'ω 4 44 JE VII.6 0.40 (HPLC-9) 413 77 eq. TFU; 1 hour 64 h 1 1 1 I o O''O ^F Y% _H F VII.7 0.42 (HPLC-9) 419 77 eq. TFU; 1 hour 65 ^f YnY, γΧ". । and i 1 1ii o Ο' Ό F VII.8 0.45 (HPLC-9) 451 77 eq. TFU; 1 hour 66 o < ^F /N °^^NH ₂ F VII.9 0.39 (HPLC-9) 429 hut TFU; 1 hour

-

Claims (18)

CLAIM 1. Compound of formula (I) where ring A is selected from the group consisting of: R ¹ is selected from the group consisting of H, F, Cl, Br, CN, -OH, C _1-4 -alkyl, -O-(C _1-4 -alkyl), -(CH ₂ ) _m COOH, -(CH ₂ ) _m -C(=O)-O-(C ₁ . ₄ -alkyl), -C(=O)-heterocyclyl, -(CH ₂ ) _m -C(=O)-NH ₂ , -(CH ₂ ) _m -C(=O)-NH-(C ₁ . ₄ alkyl), -(CH ₂ ) _m -C(=O)-N(C _1-4 -alkyl) ₂ , -C(=O)- NH-C _3-6 -cycloalkyl, -C(=O)-NH-heterocyclyl, -(CH ₂ ) _m NH-C(=O)-(C1-3-alkyl), -N( _C1-3- alkyl )-C(=O)-(C1.4-alkyl), -N(C1-3-alkyl)-C(=O)-NH2, -NH-C(=O)-NH-(C1-4alkyl) , heterocyclyl and phenyl, wherein each alkyl group or subgroup is optionally substituted with 1 or more F atoms or one OH or -O-(C _1-3 -alkyl) group; and wherein each heterocyclyl is selected from the group consisting of azetidinyl, imidazolidinyl, piperidinyl, tetrahydropyranyl and morpholinyl and is optionally substituted with one or two groups independently selected from the group consisting of oxo, C _1-3 -alkyl, -C(=O)-CH ₃ and -C(=O)-cyclopropyl; and where several R ¹ can be the same or different if n is 2; and n is an integer selected from 1 and 2; and m is an integer selected from 0, 1 and 2; and wherein in any definition mentioned above, unless otherwise indicated, any alkyl group or subgroup may be straight-chain or branched and is optionally substituted with 1 or more F atoms, or a salt thereof. 2. A compound of formula (I) according to claim 1, wherein R ¹ is selected from the group consisting of: H, F, Cl, -OH, C1-4-alkyl, -O-(C1-2-alkyl), -(C^VCOOH, -(CH ₂ )m-C(=O)-O-(C1- ₂ -alkyl), -C(=O)heterocyclyl, -(CH ₂ )mC(=O)-NH ₂ , -(CH ₂ )mC(=O)-NH-(C1.4-alkyl), -( CH2)m-C(=O)-N(CH3)(C1-3-alkyl), C(=O)-NH-cyclopropyl, -C(=O)-NH-heterocyclyl, -( _CH2 ) _m - NH-C(=O)-(C _1-3 -alkyl), -N(C _1-2 -alkyl)C(=O)-(C _1-2 -alkyl), -N(C _1-2 - alkyl)-C(=O)-NH ₂ , -NH-C(=O)-NH-(C _1-2 -alkyl), heterocyclyl and phenyl, where each alkyl group or subgroup is optionally substituted with 1 to 3 F atoms or one OH or -O-(C _1-2 -alkyl) group; and wherein each heterocyclyl is selected from the group consisting of azetidinyl, imidazolidinyl, piperidinyl, tetrahydropyranyl and morpholinyl and is optionally substituted with one or two groups independently selected from the group consisting of oxo , C _1-2 -alkyl, -C(=O)-CH ₃ and -C(=O)-cyclopropyl; and where m is 0 or 1; and where several R ¹ may be the same or different if n is 2 ; or its salt. 3. A compound of formula (I) according to claim 2, wherein R ¹ is selected from the group consisting of: H, F, -OH, -CH ₃ , -CF ₃ , -O-CH ₃ , -COOH, -(CH ₂ ) _m -C(=O)-O-CH ₃ , -(CH ₂ ) _m -C (=O)-NH ₂ , -C(=O)-NH-(C _1-3 alkyl), -(CH ₂ )-C(=O)-N(CH ₃ ) ₂ , -(CH ₂ )- C(=O)-N(CH ₃ )(CH ₂ CH ₃ ), -C(=O)-NH-cyclopropyl, 1 (cyclopropylcarbonyl)-piperidin-4-yl and 3-methyl-2-oxo-imidazolidin- 1 -yl, wherein each ethyl group or subgroup is optionally substituted at position 2 by one F atom, one OH or one -O-CH3 group; and wherein each propyl group or subgroup is optionally substituted at the 2 or 3 position by 1 to 3 F atoms; and where m means 0 or 1; and where, if n is 2, then R ¹ may be the same or different and the second R ¹ group is selected from the group consisting of F, CH ₃ , CF ₃ and phenyl; or its salt. 4. The compound of formula (I) according to claim 1, wherein ring A represents R ¹ is selected from the group consisting of H, F, -OH, C _1-4 -alkyl, -O-(C _1-4 -alkyl), -(CH ₂ ) _m -COOH, -(CH ₂ ) _m -C (=O)-O-(C ₁ . ₄ -αalkyl), -C(=O)-heterocyclyl, -(CH ₂ ) _m -C(=O)-NH ₂ , -(CH ₂ ) _m -C( =O)-NH-(C ₁ . ₄ -alkyl), -(CH ₂ ) _m -C(=O)-N(C _1-4 -alkyl) ₂ , -C(=O)-NH-C _{3 -6} -cycloalkyl, -C(=O)-NH-heterocyclyl, -(CH _{2 ) m} -NH-C(=O) -(C _1-3 -alkyl), -N(C _1.3 -alkyl) -C(=O)-(C ₁ . ₄ -alkyl), -N(C _1-3 -alkyl)-C(=O)-NH ₂ , -NH-C(=O)-NH-(C _{1 .4} -alkyl), - 62 043735 heterocyclyl and phenyl, where each alkyl group or subgroup is optionally substituted by 1 or more F atoms or one OH or -O-(C _1-3 -alkyl) group; and wherein each heterocyclyl is selected from the group consisting of azetidinyl, imidazolidinyl, piperidinyl, tetrahydropyranyl and morpholinyl and is optionally substituted with one or two groups independently selected from the group consisting of oxo, C _1-3 -alkyl, -C(=O)-CH ₃ and -C(=O)-cyclopropyl; and where several R ¹ can be the same or different if n is 2; and n is an integer selected from 1 and 2; and m is an integer selected from 0 and 1; and or its salt. 5. The compound of formula (I) according to claim 4, where R ¹ is selected from the group consisting of: H, -OH, C _1-2 -alkyl, -O-(C _1-2 -alkyl), -(CH ₂ ) _m -COOH, -(CH ₂ ) _m -C(=O)-O-(C _1-2 -alkyl), -C(=O)heterocyclyl, -(CH ₂ ) _m -C(=O)-NH ₂ , -(CH ₂ ) _m -C(=o)-NH-(C _{1- 4} -alkyl), -(CH ₂ ) _m -C(=O)-N(C _1-2 -alkyl) ₂ , -C(=O)NH-cyclopropyl, -C(=O)-NH-heterocyclyl, -(CH ₂ ) _m -NH-C(=O)-(C _1-3 -alkyl), -N(CH ₃ )-C(=O)-(C _1-2 -alkyl), -N(CH ₃ )-C(=O)-NH2, -NH-C(=O)-NH-(C _1-3 -alkyl), heterocyclyl and phenyl, where each alkyl group or subgroup is optionally substituted with 1 to 3 F atoms or one OH or -O-CH ₃ group; and wherein each heterocyclyl is selected from the group consisting of azetidinyl, imidazolidinyl, tetrahydropyranyl and morpholinyl and is optionally substituted with one or two groups independently selected from the group consisting of oxo, C _1-3 -alkyl and -C(=O)-CH ₃ ; and where, if n is 2, then R ¹ may be the same or different, the second R ¹ group is selected from the group consisting of CH3, CF3 and phenyl; or its salt. 6. The compound of formula (I) according to claim 1, wherein ring A represents R ¹ is selected from the group consisting of H, F, Cl, -OH, -O-(C _1-4 -alkyl), -C(=O)-heterocyclyl, -(CH ₂ ) _m C(=O)-NH ₂ , -(CH2)m-C(=O)-NH-(C1-4-alkyl), -(CH2)mC(=O)-N(C1-4-alkyl)2, -(CH2)m- NH-C(=O)-(C1-3-alkyl) and -N( _C1-3 -alkyl)-C(=O)-( _C1-4 -alkyl), where each alkyl group or subgroup is optionally substituted 1 or more F atoms or one OH or -O-(C _1-3 -alkyl) group; and wherein each heterocyclyl is selected from the group consisting of azetidinyl, imidazolidinyl, piperidinyl, tetrahydropyranyl and morpholinyl and is optionally substituted with one oxo or C _1-3 alkyl group; and where, if n is 2, then R ¹ may be the same or different, and the second group R is F; and n is an integer selected from 1 and 2; and m is an integer selected from 0 and 1; and or its salt. 7. The compound of formula (I) according to claim 1, wherein ring A represents R ¹ is selected from the group consisting of H, -(CH ₂ ) _m -COOH, -(CH ₂ ) _m -C(=O)-O-(C _1-4 -alkyl), -C(=O)heterocyclyl, -(CH ₂ ) _m -C(=O)-NH ₂ , -(CH ₂ ) _m -C(=O)-NH-(C _1-4 -alkyl) and -(CH ₂ ) _m -C(= O)-N(C _1-4 -alkyl) ₂ wherein each alkyl group or subgroup is optionally substituted by 1 or more F atoms or one OH or -O-(C 1-3 -alkyl) group; and wherein each heterocyclyl is selected from the group consisting of azetidinyl, imidazolidinyl, piperidinyl, tetrahydropyranyl and morpholinyl and is optionally substituted with one oxo or C _1-3 alkyl group; and where several R ¹ can be the same or different if n is 2; and n is 1; and m is an integer selected from 0 and 1; and or its salt. 8. The compound of formula (I) according to claim 1, wherein ring A represents - 63 043735 R ¹ is selected from the group consisting of H, F, Cl, Br, CN, -OH, C _1-4 -alkyl, -O-(C _1-4 -alkyl), -C(=O)-NH2, -C(=O)-NH-(C ₁ _ ₄ -alkyl), -C(=O)-N(C _1-4 -alkyl) ₂ and heterocyclyl, where each alkyl group or subgroup is optionally substituted by 1 or more atoms F or one OH or -O-(C ₁ - ₃ -alkyl) group; and wherein each heterocyclyl is selected from the group consisting of azetidinyl and piperidinyl, and is optionally substituted with one C _1-3 alkyl, -C(=O)-CH ₃ or -C(=O)-cyclopropyl group; and where, if n is 2, then R ¹ may be the same or different and the second group R ¹ is selected from the group consisting of F and CH ₃ ; and n is an integer selected from 1 and 2; and or its salt. 9. A compound of formula (I) according to claim 1, selected from the group consisting of: - 64 043735 - 65 043735 or its salt. 12. A compound of formula (I) according to claim 9, having the structure: 13. A compound of formula (I) according to claim 9, having the structure: 14. A compound of formula (I) according to claim 9, having the structure: 15. A compound of formula (I) according to claim 9, having the structure: - 66 043735 16. A pharmaceutically acceptable salt of the compound according to any one of p. 1-15. 17. Use of a compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof according to claim 16 as a drug that inhibits the activity of AOSZ. 18. Use of a compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof according to claim 16 for the treatment of cancer, NASH (non-alcoholic steatohepatitis), pulmonary fibrosis, retinopathy, nephropathy or stroke. 19. A pharmaceutical composition containing a therapeutically effective amount of a compound according to any one of claims 1 to 15 or a pharmaceutically acceptable salt thereof according to claim 16, and together with one or more inert carriers and/or diluents. 20. A method of treating a disease or condition that is mediated by inhibition of AOSZ activity, comprising administering a therapeutically effective amount of a compound according to any one of claims 115 or a pharmaceutically acceptable π salt thereof. 16 to the patient in need. Eurasian Patent Organization, EAPO Russia, 109012, Moscow, Maly Cherkassky lane, 2