EA045055B1 - 2-FORMYL-3-HYDROXYPHENYLOXYMETHYL COMPOUNDS AS A HEMOGLOBIN MODULATOR - Google Patents

Description

Cross references to related applications

This application claims priority under 35 U.S. Code §119(e) to U.S. Provisional Application No. 62/769,196, filed November 19, 2018, and U.S. Provisional Application No. 62/821,314, filed March 20, 2019. ., U.S. Provisional Patent Application No. 62/848,773, filed May 16, 2019, and U.S. Provisional Patent Application No. 62/883,313, filed Aug. 6, 2019, the contents of each of which are incorporated herein by reference in their entirety.

Field of technology

The present application provides compounds and pharmaceutical compositions suitable as hemoglobin modulators and methods of using them for the treatment of hemoglobin-mediated disorders.

State of the art

Sickle cell disease is a disease of the red blood cells, particularly common in people of African and Mediterranean descent. Sickle cell disease is based on sickled hemoglobin (HbS), which contains a point mutation relative to the predominant peptide sequence of hemoglobin type A (HbA).

Hemoglobin (Hb) carries oxygen molecules from the lungs to various tissues and organs throughout the body. Hemoglobin binds and releases oxygen through conformational changes. Sickle hemoglobin (HbS) contains a point mutation that replaces glutamic acid with valine, making sickle hemoglobin susceptible to polymerization under hypoxic conditions, giving sickle hemoglobin-containing red blood cells their characteristic sickle shape. Sickle cells are also stiffer than normal red blood cells, and their inherent lack of flexibility can lead to clogged blood vessels.

2-G hydroxy-6-((2-(1-isopropyl-1H-nirazol-5-yl)pyridin-3-yl)methoxy)benzaldehyde (also known as voxelotor or GBT440), a hemoglobin modulator that increases the affinity of hemoglobin to oxygen and therefore inhibits the polymerization of sickle hemoglobin under hypoxic conditions, is currently in phase 3 clinical trials for the treatment of sickle cell disease (NCT03036813).

WO 2014/150268 describes hemoglobin modulators that are structurally related to the compounds disclosed herein.

There is a need for compounds that are capable of treating disorders mediated by abnormal hemoglobin, such as sickle hemoglobin, and methods for treating such disorders. Of particular interest are compounds that have an improved pharmacokinetic profile compared to known hemoglobin modulators while maintaining or improving efficacy, as such compounds will allow favorable dosing schedules (eg, lower and/or less frequent dosing).

The essence of the invention

The present description provides a compound of formula I:

I, or an isotopically enriched analogue, stereoisomer, mixture of stereoisomers or prodrug thereof, or a pharmaceutically acceptable salt of each, wherein:

X represents CH or N;

Y represents CH or N;

Z is absent or CH2, O or S; and

R ¹ represents monohydroxy-(C _1-4 alkyl), dihydroxy-(C _1-4 alkyl), -CH ₂ CH ₂ OCH ₃ , -CH2CH2CN or

Some embodiments of the invention provide pharmaceutical compositions containing a compound as defined herein, or an isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug, or a pharmaceutically acceptable salt of each thereof, and a pharmaceutically acceptable excipient. Some embodiments of the invention relate to pharmaceutical compositions containing a compound as defined herein, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. Some embodiments of the invention relate to pharmaceutical compositions containing a compound as defined herein and a pharmaceutically acceptable excipient.

Also provided herein are methods of increasing the oxygen affinity of hemoglobin (eg, hemoglobin type S) in a subject in need thereof, comprising administering to the subject a compound as defined herein or a pharmaceutical composition as defined herein.

Also provided herein are methods of treating a hemoglobin-mediated disorder in a subject in need thereof, comprising administering to the subject a compound as defined herein or a pharmaceutical composition as defined herein.

Also provided herein are methods of treating sickle cell disease in a subject in need thereof, comprising administering to the subject a compound as defined herein or a pharmaceutical composition as defined herein.

Detailed description of the invention

Definitions

As used herein, the following words, phrases and symbols generally have the meanings set forth below, unless the context in which they are used indicates otherwise.

A prime (-) that is not between two letters or symbols is used to indicate the point of attachment of a substituent. For example, -C(O)NH2 is attached via a carbon atom. A prime at the beginning or end of a chemical group is a matter of convenience; chemical groups can be depicted with or without one or more strokes without losing their normal meaning. A wavy line or a dotted line drawn through or perpendicular to the end of a line in a structure indicates a specified group attachment point. Unless chemically or structurally necessary, neither directionality nor stereochemistry is indicated or implied by the order in which a chemical group is written or named.

The prefix C _u - _v indicates that the following group has from and to v carbon atoms. For example, C1_ ₆ alkyl indicates that the alkyl group has 1 to 6 carbon atoms. In another example, C ₁ . ₄ αlkyl indicates that the alkyl group has 1 to 4 carbon atoms.

Reference to an exemplary value or parameter in the present specification includes (and describes) embodiments of the invention that are directed to the given value or parameter as such. In some embodiments, the term approximately includes the indicated amount ±10%. In other embodiments, the term approximately includes the indicated amount ±5%. In some other embodiments, the term approximately includes the indicated amount ±1%. Also, the term about x includes a description of x. In addition, the singular forms a and the include references to plural forms unless the context clearly indicates otherwise. Thus, for example, a reference to a compound includes a plurality of such compounds, and a reference to an assay includes a reference to one or more assays and their equivalents known to those skilled in the art.

Alkyl refers to a straight or branched saturated hydrocarbon chain. As used herein, alkyl has 1 to 20 carbon atoms (i.e. C1_ ₂₀ alkyl), 1 to 12 carbon atoms (i.e. C1_ ₁₂ alkyl), 1 to 8 carbon atoms (i.e. C _{1 8} alkyl), 1 to 6 carbon atoms (i.e. C _{1 6} alkyl) or 1 to 4 carbon atoms (i.e. C1_ ₄ alkyl). Examples of alkyl groups include, for example, methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, pentyl, 2-pentyl, isopentyl, neopentyl, hexyl, 2-hexyl, 3-hexyl and 3 -methylpentyl. When an alkyl moiety having a specific number of carbon atoms is named by a chemical name or identified by a molecular formula, all positional isomers having that number of carbon atoms may be included; thus, for example, butyl includes n-butyl (i.e. -(CH ₂ ) ₃ CH ₃ ), sec-butyl (i.e. CH(CH ₃ )CH ₂ CH ₃ ), isobutyl (i.e. -CH ₂ CH(CH ₃ ) ₂ ) and tert-butyl (i.e. -C(CH ₃ ) ₃ ); and propyl includes npropyl (ie -(CH2CH3) and isopropyl (ie -CH(CH3) ₂ ).

Some commonly used alternative chemical names may be used. For example, a divalent group such as a divalent alkyl group, a divalent aryl group, etc. may also be called an alkylene group or an alkylenyl group, an arylene group or an arylenyl group, respectively. In addition, unless explicitly stated otherwise, when combinations of groups are referred to herein as a single moiety, such as arylalkyl or aralkyl, the last mentioned group contains the atom by which the moiety is attached to the rest of the molecule.

Alkenyl refers to an alkyl group containing at least one carbon-carbon double bond and having from 2 to 20 carbon atoms (i.e. C ₂ - ₂₀ alkenyl), from 2 to 8 carbon atoms

- 2 045055 (i.e. C ₂ - ₈ alkenyl), from 2 to 6 carbon atoms (i.e. C _2-6 alkenyl) or from 2 to 4 carbon atoms (i.e. C ₂ - ₄ alkenyl) . Examples of alkenyl groups include, for example, ethenyl, propenyl, butadienyl (including 1,2-butadienyl and 1,3-butadienyl).

Alkynyl refers to an alkyl group containing at least one carbon-carbon triple bond and having 2 to 20 carbon atoms (i.e. _C2-20 alkynyl), 2 to 8 carbon atoms (i.e. _C2-8 alkynyl ), from 2 to 6 carbon atoms (ie C _2-6 alkynyl) or from 2 to 4 carbon atoms (ie C _2-4 alkynyl). The term alkynyl also includes groups having one triple bond and one double bond.

Alkoxy belongs to the alkyl-O-type group. Examples of alkoxy groups include, for example, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, t-butoxy, sec-butoxy, n-pentoxy, n-hexoxy and 1,2-dimethylbutoxy.

Alkylthio belongs to the alkyl-S-type group. Alkylsulfinyl belongs to the alkyl-S(O)- type group. Alkylsulfonyl belongs to the alkyl-S(O) ₂ - type group.

Acyl refers to a group of the type -C(O)R ^y , where R ^y is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein.

Examples of acyl include, for example, formyl, acetyl, cyclohexylcarbonyl, cyclohexylmethylcarbonyl and benzoyl.

Amido refers to both a C-amido type group, which refers to a -C(O)NR ^y R ^z type group, and an N-amido type group, which refers to a -NR ^y C(O)R ^z type group, where R ^y and R ^z are independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein, or R ^y and R ^z taken together to form cycloalkyl or heterocyclyl; each of which may be optionally substituted as defined herein.

Amino refers to a group of the type -NR ^y R ^z where R ^y and R ^z are independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein.

Amidino refers to -C(NR ^y )(NR ^z ₂ ), wherein R ^y and R ^z are independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein.

Aryl refers to an aromatic carbocyclic group having one ring (eg, monocyclic) or multiple rings (eg, bicyclic or tricyclic), including fused systems. As used herein _{, aryl has 6 to 20 ring carbons (e.g., C6-20 aryl} ), 6 to 12 ring carbons (e.g., _C6-12 aryl), or 6 to ₁₀ ring carbons ( for example, C ₆ - ₁₀ aryl). Examples of aryl groups include, for example, phenyl, naphthyl, fluorenyl and anthryl. Aryl, however, does not in any way include or overlap with heteroaryl, as defined below. If one or more aryl groups are fused to a heteroaryl, the resulting ring system is heteroaryl regardless of the point of attachment. If one or more aryl groups are fused to a heterocyclyl, the resulting ring system is heterocyclyl regardless of the point of attachment.

Arilalkyl or aralkyl refers to the aryl-alkyl- group.

Carbamoyl refers to both the O-carbamoyl group, which is a -OC(O)NR ^y R ^z type group, and the N-carbamoyl group, which is a -NR ^y C(O)OR ^z type group, where R ^y and R ^z independently represent hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein.

Carboxyl ether or ester refers to both -OC(O)R ^x and -C(O)OR ^x , where R ^x is alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein.

Cycloalkyl refers to a saturated or partially unsaturated cyclic alkyl group having one or more rings, including fused, bridged and spiro ring systems. The term cycloalkyl includes cycloalkenyl groups (i.e. a cyclic group having at least one double bond) and carbocyclic fused ring systems containing at least one carbon atom in sp ³ hybridization (i.e. at least one non-aromatic ring) . As used herein, cycloalkyl has from 3 to ₂₀ ring carbon atoms (i.e., C ₃ to 20 cycloalkyl), from 3 to 12 ring carbon atoms (i.e., C ₃ to ₁₂ cycloalkyl), from 3 to 10 carbon atoms in ring (eg, C ₃ - ₁₀ cycloalkyl), 3 to 8 carbon atoms in the ring (eg, C _{3 - 8} cycloalkyl) or 3 to 6 carbon atoms in the ring (eg, C ₃ - ₆ cycloalkyl). Monocyclic groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. Polycyclic groups include, for example, bicyclo[2.2.1]heptanyl, bicyclo[2.2.2]octanyl, adamantyl, norbornyl, decalinyl, 7,7-dimethylbicyclo[2.2.1]heptanyl and the like. Additionally, the term cycloalkyl is intended to include any non-aromatic ring that can be fused to an aryl ring, regardless of attachment to the rest of the molecule. In addition, cycloalkyl also includes spirocycloalkyl when there are two positions for substitution on the same carbon atom, for example spiro[2,5]octanyl, spiro[4.5]decanyl or spiro[5.5]undecanyl.

Cycloalkylalkyl refers to a group of the cycloalkyl-alkyl- type.

Guanidino refers to -NR ^y C(=NR ^z )(NR ^y R ^z ), wherein R ^y and R ^z are each independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein.

Imino refers to a group of the type -C(NR ^y )R ^z wherein R ^y and R ^z are each independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein.

Imido refers to a group of the type -C(O)NR ^y C(O)R ^z wherein R ^y and R ^z are each independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein.

Halogen or halogeno refers to atoms occupying group VIIA of the periodic table, such as fluorine, chlorine, bromine or iodine.

Haloalkyl refers to a straight or branched alkyl group, as defined above, where one or more (eg, 1 to 6 or 1 to 3) hydrogen atoms are replaced by halogen. For example, if a residue is substituted with more than one halogen, it can be referred to by a prefix corresponding to the number of halogen moieties attached. Dihaloalkyl and trihaloalkyl refer to alkyl substituted with two (di) or three (three) halogen-containing groups, which may, but is not necessarily, the same halogen. Examples of haloalkyl include, for example, trifluoromethyl, difluoromethyl, fluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl and the like.

Haloalkoxy refers to an alkoxy group, as defined above, where one or more (eg, 1 to 6 or 1 to 3) hydrogen atoms are replaced by halogen.

Hydroxyalkyl refers to an alkyl group, as defined above, where one or more (eg, 1 to 6 or 1 to 3) hydrogen atoms are replaced by a hydroxy group. _{Monohydroxy-(C1_4alkyl )} refers to _C1 . ₄ alkyl group, as defined above, in which one hydrogen atom is replaced by a hydroxy group. Dihydroxy-(C _{1_4} alkyl) refers to C _{1 . 4} alkyl group, as defined above, in which two hydrogen atoms are replaced by hydroxy groups.

Heteroalkyl refers to an alkyl group in which one or more carbon atoms (and any associated hydrogen atoms) are independently replaced by a similar or different heteroatom group, provided that the point of attachment to the rest of the molecule is through a carbon atom. The term heteroalkyl includes a straight or branched saturated chain containing carbon and heteroatoms. For example, 1, 2 or 3 carbon atoms may be independently replaced by a similar or different heteroatom group. Heteroatomic groups include, but are not limited to, -NR ^y -, -O-, -S-, -S(O)-, -S(O)2- and the like, where R ^y represents hydrogen, alkyl , alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein. Examples of heteroalkyl groups include, for example, ethers (for example, -CH2OCH3, -CH(CH3) _OCH3 , _{-CH2CH2OCH3 , -CH2CH2OCH2CH2OCH3 ,} etc.), _thioethers (for example, - CH2SCH3, -CH(CH ₃ )SCH ₃ , -CH2CH2SCH3, -CH ₂ CH ₂ SCH ₂ CH ₂ SCH ₃ , etc.), sulfones (for example, -CH ₂ S(O) ₂ CH ₃ , -CH( CH ₃ )S(O) ₂ CH ₃ , -CH2CH2S(O)2CH3, -CH2CH2S(O)2CH2CH2OCH3, etc.) and amines (for example, -CHNRyCB, -CH(CH3)NR ^y CH3, -CH2CH2NR ^y CH3, -CHCHNRyC^CHNRyCH, etc., wherein R ^y is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl, each of which may be optionally substituted as defined herein). As used herein, heteroalkyl includes 1 to 10 carbon atoms, 1 to 8 carbon atoms, or 1 to 4 carbon atoms; and 1 to 3 heteroatoms, 1 to 2 heteroatoms, or 1 heteroatom.

Heteroaryl refers to an aromatic group having a single ring, multiple rings or multiple fused rings, with one or more ring heteroatoms independently selected from nitrogen, oxygen and sulfur. As used herein, heteroaryl has 1 to 20 ring carbons (e.g., _{C1_20} heteroaryl), 3 to 12 ring carbons (e.g., _C3-12 heteroaryl), or 3 to 8 ring carbons (e.g., C ₃ - ₈ heteroaryl), and 1 to 5 heteroatoms per ring, 1 to 4 heteroatoms per ring, 1 to 3 heteroatoms per ring, 1 to 2 heteroatoms per ring, or 1 heteroatom per ring, independently selected from nitrogen, oxygen and sulfur. In some cases, heteroaryl includes 5-10 membered ring systems, 5-7 membered ring systems, or 56-membered ring systems, each independently containing from 1 to 4 heteroatoms per ring, from 1 to 3 heteroatoms per ring, from 1 up to 2 heteroatoms per ring or 1 ring heteroatom independently selected from nitrogen, oxygen and sulfur. Examples of heteroaryl groups include, for example, acridinyl, benzimidazolyl, benzothiazolyl, benzidolyl, benzofuranyl, benzothiazolyl, benzothiadiazolyl, benzonaphthofuranyl, benzoxazolyl, benzothienyl (benzothiophenyl), benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridyl, carbazolyl, cinnolinyl, dibenzofuranyl, dibenzothiophenyl, furanyl,

- 4 045055 isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, isoquinolyl, isoxazolyl, naphthyridinyl, oxadiazolyl, oxazolyl, 1-oxidopyridinyl, 1-oxidopyrimidinyl, 1-oxidopyridazinyl, 1oxidopyridazinyl, phenazinyl, phthalazinyl, pteridine yl, purinyl, pyrrolyl, pyrazolyl , pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl and triazinyl. Examples of fused heteroaryl rings include, but are not limited to, benzo[d]thiazolyl, quinolinyl, isoquinolinyl, benzo[b]thiophenyl, indazolyl, benzo[d]imidazolyl, pyrazolo[1,5-a]pyridinyl and imidazo[1,5a ]pyridinyl, where the heteroaryl can be linked through any ring of the fused system. Any aromatic ring having one or more fused rings containing at least one heteroatom is considered a heteroaryl regardless of attachment to the rest of the molecule (ie, through any of the fused rings). Heteroaryl does not include or overlap with aryl as defined above.

Heteroarylalkyl refers to a group of the heteroaryl-alkyl- type.

Heterocyclyl refers to a saturated or partially unsaturated cyclic alkyl group with one or more ring heteroatoms independently selected from nitrogen, oxygen and sulfur. The term heterocyclyl includes heterocycloalkenyl groups (ie, a heterocyclyl group having at least one double bond), bridged heterocyclyl groups, fused heterocyclyl groups, and spiroheterocyclyl groups. The heterocyclyl may be a single ring or multiple rings, where multiple rings may be fused, bridged, or spiro, and may contain one or more (e.g., 1 to 3) oxo (=O) or N-oxide (-O-) fragments. Any non-aromatic ring containing at least one heteroatom is considered a heterocyclyl, regardless of attachment (ie, may be bonded through a carbon atom or a heteroatom). In addition, the term heterocyclyl is intended to include any non-aromatic ring containing at least one heteroatom, which ring may be fused to an aryl or heteroaryl ring, regardless of attachment to the rest of the molecule. As used herein, a heterocyclyl has from 2 to 20 carbon atoms in the ring (i.e. heterocyclyl C ₂ . ₂ o), _from 2 to 12 carbon atoms in the ring (i.e. heterocyclyl C _{2_12} ), from 2 to 10 carbon atoms in the ring (i.e. heterocyclyl C ₂ - ₁₀ ), from 2 to 8 carbon atoms in the ring (for example, C _{2_8} heterocyclyl ₎ , from 3 to ₁₂ carbon atoms in the ring (for example, C _{3_12} heterocyclyl ), from ₃ to 8 carbon _atoms in the ring (for example, C _{3_8} heterocyclyl) or from 3 to 6 carbon atoms in the ring (i.e. C _{3_6} heterocyclyl); having from 1 to 5 heteroatoms in the ring, from 1 to 4 heteroatoms in the ring, from 1 to 3 heteroatoms in the ring, from 1 to 2 heteroatoms in the ring, or 1 heteroatom in the ring, independently selected from nitrogen, sulfur or oxygen. Examples of heterocyclyl groups include, for example, azetidinyl, azepinyl, benzodioxolyl, benzo[b][1,4]dioxepinyl, 1,4-benzodioxanyl, benzopyranyl, benzodioxynyl, benzopyranonyl, benzofuranonyl, dioxolanyl, dihydropyranyl, hydropyranyl, thienyl[1,3] dithianyl, decahydroisoquinolyl, furanonyl, imidazolinyl, imidazolidinyl, indolinyl, indolizinyl, isoindolinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl , oxazolidinyl, oxiranil, oxetanil, phenothiazinyl, phenoxazinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, tetrahydropyranyl, tritianyl, tetrahydroquinolinyl, thiophenyl (i.e. thienyl), tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1oxothiomorpholinyl and 1 ,1-dioxothiomorpholinyl. The term heterocyclyl also includes spiroheterocyclyl when there are two substitution positions on the same carbon atom. Examples of spiroheterocyclyl rings include, for example, bicyclic and tricyclic ring systems such as 2-oxa-7-azaspiro[3.5]nonanyl, 2-oxa-6-azaspiro[3.4]octanyl and 6-oxa-1-azaspiro[3.3]heptanyl . Examples of fused heterocyclyl rings include, but are not limited to, 1,2,3,4-tetrahydroisoquinolinyl, 4,5,6,7-tetrahydrothieno[2,3-c]pyridinyl, indolinyl and isoindolinyl, where the heterocyclyl may be linked through any ring of a condensed system.

Heterocyclylalkyl refers to a group of the heterocyclyl-alkyl- type.

Oxime refers to a group of the type -CR ^y (=NOH), where R ^y represents hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein.

Sulfonyl refers to a group of the type -S(O) ₂ R ^y , where R ^y represents hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein. Examples of sulfonyl are methylsulfonyl, ethylsulfonyl, phenylsulfonyl and toluenesulfonyl.

Sulfinyl refers to a group of the type -S(O)R ^y , where R ^y represents hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl; each of which may be optionally substituted as defined herein. Examples of sulfinyl are methylsulfinyl, ethylsulfinyl, phenylsulfinyl and toluenesulfinyl.

Sulfonamido refers to groups of the type -SO ₂ NR ^y R ^z and -NR ^y SO ₂ R ^z , wherein R ^y and R ^z are each independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroalkyl or heteroaryl;

- 5 045055 each of which may be optionally substituted as defined herein.

The terms optional or optional mean that the event or circumstance described below may or may not occur, and that the description includes cases in which the event or circumstance described occurs and cases in which it does not occur. Also, the term optionally substituted refers to any one or more (eg, 1 to 5 or 1 to 3) hydrogen atoms on the designated atom or group, which may or may not be replaced by a moiety other than hydrogen.

The term substituted as used herein means any of the above groups (i.e. alkyl, alkenyl, alkynyl, alkylene, alkoxy, haloalkyl, haloalkoxy, cycloalkyl, aryl, heterocyclyl, heteroaryl and/or heteroalkyl), wherein at least one (e.g., 1 to 5 or 1 to 3) the hydrogen atom is replaced by a bond with an atom other than hydrogen, such as, but not limited to, alkyl, alkenyl, alkynyl, alkoxy, alkylthio, acyl, amido, amino, amidino aril, arail, azido, carbamil, carboxil, carbon acid, cycloal, cycloalkil, guanidino, halo, halogenalkil, haloenalcoxial, hydroxial, heteroaril, heteroarilalkil, heterocycliclles, and heterocyclicyl. Nh2, = nnh2, image, image , hydroxy, oxo, oxime, nitro, sulfonyl, sulfinyl, alkylsulfonyl, alkylsulfinyl, thiocyanate, -S(O)OH, -S(O)2OH, sulfonamido, thiol, thioxo, Noxide or -Si(R ^y ) ₃ , where each R ^y independently represents hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, aryl, heteroaryl or heterocyclyl.

In some embodiments, substituted includes any of the above alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl groups in which one or more (e.g., 1 to 5 or 1 to 3) hydrogen atoms are replaced by deuterium, halogen , cyano, nitro, azido, oxo, alkyl, alkenyl, alkynyl, haloalkyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -NR ^g R ^h , -NR ^g C(=O)R ^h , -NR ^g C(=O) NR ^g R ^h , -NR ^g C(=O)OR ^h , -NR ^g S(=O)1-2R ^h , -C(=O)R ^g -C(=O)OR ^g , -OC(= O)OR ^g -OC(=O)R ^g , -C(=O)NR ^g R ^h , -OC(=O)NR ^g R ^h , -OR ^g , -SRS -S(=O)R ^g , -S(=O)2R ^g , -OS(=O)1-2R ^g , -S(=O)1-2OR ^g , -NR ^g S(=O)1-2NR ^g R ^h , =NSO2R ^g , =NOR ^g , -S(=O)1-2NR ^g R ^h , -SF5, -SCF3 or -OCF3. In some embodiments, substituted also means any of the above groups in which one or more (e.g., 1 to 5 or 1 to 3) hydrogen atoms are replaced by -C(=O)R ^g , -C(=O) OR ^g , -C(=O)NR ^g R ^h , -CH2SO ₂ R ^g , or -CH ₂ SO ₂ NR ^g R ^h . In the above, R ^g and R ^h are similar or different and independent of each other: hydrogen, alkyl, alkenyl, alkynyl, alkoxy, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclyl, heterocyclylalkyl, heteroaryl and/or heteroarylalkyl. In some embodiments, substituted also means any of the above groups in which one or more (e.g., 1 to 5 or 1 to 3) hydrogen atoms are replaced by an amino, cyano, hydroxyl, imino, nitro, oxo, thioxo bond , halo, alkyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkylalkyl, haloalkyl, heterocyclyl, Nheterocyclyl, heterocyclylalkyl, heteroaryl and/or heteroarylalkyl, or two of R ^g and R ^h and Ri are taken together with the atoms to which they are attached to form a heterocyclyl ring, optionally substituted with oxo, halogen or alkyl, optionally substituted with oxo, halogen, amino, hydroxyl or alkoxy.

Polymers or similar undefined structures obtained by defining substituents with additional substituents attached ad infinitum (eg, a substituted aryl having a substituted alkyl that is itself substituted by a substituted aryl group that is further substituted by a substituted heteroalkyl group, etc.) are not are intended to be included herein. Unless otherwise stated, the maximum number of consecutive substitutions in the compounds described herein is three. For example, sequential substitutions of substituted aryl groups by two other substituted aryl groups are limited to ((substituted aryl)substituted aryl)substituted aryl. Likewise, the above definitions are not intended to include unacceptable types of substitution (eg, methyl substituted with 5 fluorines or heteroaryl groups having two adjacent oxygen atoms on the ring). Such unacceptable types of substitution are well known to one skilled in the art. When using a chemical group for modification, the term substituted may describe other chemical groups as defined herein.

In some embodiments, as used herein, the phrase one or more refers to the range from one to five. In some embodiments, as used herein, the phrase one or more refers to the range from one to three.

Any compound or structure described herein is intended to represent unlabeled forms as well as isotopically labeled forms (isotopologues) of the compounds. These forms of compounds may also be named and include isotopically enriched analogues. Isotopically labeled compounds have the structures depicted herein, except that one or more atoms are replaced by an atom having a given atomic mass or mass number. Examples of isotopes that may be included in the described compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine and iodine, such as ^2H , ^3H , ^nC , ^13C , ^14C ,

- 6 045055 ¹³ N, ¹⁵ N, ¹⁵ O, ¹⁷ O, ¹⁸ O, ³¹ P, ³² P, ³⁵ S, ¹⁸ F, ³⁶ Cl, ¹²³ I and ¹²⁵ I, respectively. Various isotopically labeled compounds of the present invention are possible, such as those incorporating radioactive isotopes such as ³ H, ¹³ C and ¹⁴ C. Such isotopically labeled compounds may be useful in metabolic studies, reaction kinetics studies, detection or imaging techniques, such as positron emission tomography (PET) or single photon emission computed tomography (SPECT), including assays of drug or substrate tissue distribution, or for radioactive treatment of patients.

The term isotope-enriched analogues includes deuterated analogues of the compounds described herein in which one or more hydrogen atoms are/are replaced by deuterium, for example hydrogen at the carbon atom. Such compounds exhibit increased resistance to metabolism and are thus useful for increasing the half-life of any compound when administered to a mammal, especially a human. See, for example, Foster, Deuterium Isotope Effects in Studies of Drug Metabolism, Trends Pharmacol. Sci. 5(12):524-527 (1984). Such compounds are synthesized by methods well known in the art, for example, using starting materials in which one or more hydrogen atoms are replaced by deuterium.

Deuterium-labeled or substituted therapeutic compounds of the present invention may have improved DMPK (drug metabolism and pharmacokinetics) properties related to distribution, metabolism and elimination (ADME). Substitution with heavier isotopes such as deuterium may provide certain therapeutic benefits due to greater metabolic stability, such as increased in vivo half-life, reduced dosage requirements, and/or improved therapeutic index. The compound labeled with ¹⁸ F, ³ H, ^PS can be used for PET, SPECT or other imaging studies. The isotopically labeled compounds of the present invention and their prodrugs can generally be prepared by following the procedures disclosed in the Schemes or in the Examples and Preparations described below by replacing the readily available non-isotopically labeled reagent with an isotopically labeled reagent. It is obvious that deuterium in this context is considered as a substituent in the compound specified herein.

The concentration of such a heavier isotope, specifically deuterium, can be determined by means of an isotope enrichment factor. In the compounds of the present invention, any atom not expressly designated as a specific isotope means any stable isotope of that atom. Unless otherwise stated, when a position is specifically designated as H or hydrogen, it is understood that the position contains hydrogen in its natural isotopic composition in abundance. Accordingly, in the compounds of the present invention, any atom specifically designated as deuterium (D) means deuterium. Additionally, in some embodiments, a corresponding deuterated analogue is provided.

In many cases, the compounds of the present invention are capable of forming acidic and/or basic salts due to the presence of amino and/or carboxyl groups or the like.

Also provided are a pharmaceutically acceptable salt, an isotopically enriched analogue, a deuterated analogue, an isomer (such as a stereoisomer), a mixture of isomers (for example, a mixture of stereoisomers), a prodrug and a metabolite of the compounds described herein.

Pharmaceutically acceptable or physiologically acceptable refers to compounds, salts, compositions, dosage forms and other materials that can be used to prepare a pharmaceutical composition suitable for veterinary or human pharmaceutical use.

The term pharmaceutically acceptable salt of a given compound refers to salts that retain the biological effectiveness and properties of the compound and that are not biologically or otherwise undesirable. Pharmaceutically acceptable salts or physiologically acceptable salts include, for example, inorganic acid salts and organic acid salts. In addition, when the compounds described herein are obtained as an acid addition salt, the free base can be prepared by making the acid salt solution alkaline. Conversely, if the product is a free base, an addition salt, particularly a pharmaceutically acceptable addition salt, can be prepared by dissolving the free base in a suitable organic solvent and treating the solution with an acid according to standard procedures for preparing acid addition salts from basic compounds. Various synthetic techniques are known to those skilled in the art that can be used to prepare non-toxic pharmaceutically acceptable addition salts. Pharmaceutically acceptable acid addition salts can be prepared from inorganic and organic acids. Salts derived from inorganic acids include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like. Salts derived from organic acids include, for example, acetic acid, propionic acid, gluconic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic

- 7 045055 acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, phenylacrylic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, etc. Likewise, pharmaceutically acceptable base addition salts can be prepared from inorganic and organic bases. Salts derived from non-limiting bases include, by way of example only, sodium, potassium, lithium, aluminum, ammonium, calcium and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary and tertiary amines such as alkylamines (i.e. NH ₂ (alkyl)), dialkylamines (i.e. NH (alkyl) ₂ ), trialkylamines (i.e. N(alkyl) ₃ ), substituted alkylamines (i.e. NH ₂ (substituted alkyl)), di(substituted alkyl) amines (i.e. HN(substituted alkyl) ₂ ), tri(substituted alkyl)amines (i.e. K(substituted alkyl) ₃ ), alkenyl amines (i.e. NH ₂ (alkenyl)), dialkenyl amines (i.e. HN(alkenyl) ₂ ), trialkenyl amines (i.e. N( alkenyl) ₃ ), substituted alkenyl amines (i.e. NH ₂ (substituted alkenyl)), di(substituted alkenyl) amines (i.e. NH (substituted alkenyl) ₂ ), tri(substituted alkenyl) amines (i.e. ^substituted alkenyl) ₃ , mono-, di-or tricycloalkylamines (i.e. NH ₂ (cycloalkyl), HN(cycloalkyl) ₂ , N(cycloalkyl) ₃ ), mono-, di- or triarylamines (e.g. NH ₂ (aryl) , HN(aryl) ₂ , N(αryl) ₃ ) or mixed amines, etc. Specific examples of suitable amines include, by way of example only, isopropylamine, trimethylamine, diethylamine, tri(isopropyl)amine, tri(n-propyl)amine, ethanolamine, 2-dimethylaminoethanol, piperazine, piperidine, morpholine, N-ethylpiperidine, and the like. . In some embodiments, the pharmaceutically acceptable salt does not contain a primary amine salt.

The term hydrate refers to a complex formed by combining a compound as defined herein and water.

Solvate refers to the association or complex of one or more solvent molecules and a compound of the present invention. Examples of solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid and ethanolamine.

Some of the compounds exist as tautomers. Tautomers are in equilibrium with each other. For example, amide-containing compounds can exist in equilibrium with imido acid tautomers. Regardless of which tautomer is shown, and regardless of the nature of the equilibrium between tautomers, one skilled in the art will understand that the compounds include both amide and imido acid tautomers. Thus, it is intended that amide-containing compounds include imido acid tautomers. Likewise, imido acid-containing compounds are intended to include amide tautomers.

The compounds of the present invention or their pharmaceutically acceptable salts include an asymmetric center and thus can give enantiomers, diastereomers and other stereoisomeric forms, which can be defined in terms of absolute stereochemistry as (R)- or (S)- or as (D ) or (L) for amino acids. The present invention includes all such possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (-), (R)- and (S)- or (D)- and (L)-isomers can be prepared using chiral synthons or chiral reagents, or can be separated using standard methods, e.g. chromatography and fractional crystallization. Standard methods for preparing/isolating individual enantiomers include chiral synthesis from a suitable optically pure precursor or resolution of the racemate (or racemate salt or derivative) using, for example, chiral high performance liquid chromatography (HPLC). When compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, and unless otherwise indicated, the compounds are intended to include type E and Z geometric isomers.

A stereoisomer refers to a compound that is made up of the same atoms, linked through the same bonds, but has different three-dimensional structures that are not interchangeable. The present invention contemplates various stereoisomers and mixtures thereof and includes enantiomers, which refer to two stereoisomers whose molecules are non-superimposed mirror images of each other.

Diastereomers are stereoisomers that have at least two asymmetric atoms but are not mirror images of each other.

The relative centers of compounds as depicted herein are indicated graphically using a thick bond style (thick or parallel lines) and absolute stereochemistry is depicted using wedge bonds (thick or parallel lines).

Prodrug means any compound that releases in vivo the active parent drug according to the structure defined herein when such prodrug is administered to a mammalian subject. Prodrugs of a compound provided herein are prepared by modifying functional groups present in a compound provided herein such that the modifications can be cleaved in vivo to release the parent compound. Prodrugs can be prepared by modifying functional groups present in compounds such that the modifications are cleaved to the parent compounds.

- 8 045055 nii either during routine manipulations or in vivo. Prodrugs include compounds as defined herein in which the hydroxy, amino, carboxyl or sulfhydryl group in the compound as defined herein is linked to any group that can be cleaved in vivo to regenerate a free hydroxy, amino or sulfhydryl group respectively. Examples of prodrugs include, but are not limited to, esters (e.g., acetate, formate, and benzoate derivatives), amides, guanidines, carbamates (e.g., N,N-dimethylaminocarbonyl) of hydroxyl functional groups in the compounds described herein, etc. P. The preparation, selection and use of prodrugs are discussed in T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, Vol. 14 of the A.C.S. Symposium Series; Design of Products, ed. H. Bundgaard, Elsevier, 1985; and in Bioreversible Carriers in Drug Design, ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987, the contents of each of which are incorporated herein by reference in their entirety.

The term metabolite as used herein refers to the product resulting from the metabolism of a compound as defined herein. As used herein, the term metabolized refers to the sum of the processes (including, but not limited to, hydrolysis reactions and enzyme-catalyzed reactions) by which a specific substance, such as a compound described herein, is changed by the body. For example, the aldehyde moiety of the (-C(O)H) type of the compounds of the present invention can be reduced in vivo to a moiety of the -CH2OH type.

The term hydroxy protecting group refers to a chemical moiety that is added to and then removed from hydroxyl functionality to obtain chemoselectivity in a subsequent chemical reaction. Examples of protecting groups as well as deprotection methods include, but are not limited to, acetyl (Ac) (removed by acid or base), benzoyl (Bz) (removed by acid or base), benzyl (Bn) (removed by hydrogenolysis ), β-methoxyethoxymethyl ether (MEM) (removed with acid), dimethoxytrityl or [bis-(4-methoxyphenyl)phenylmethyl](DMT) (removed with weak acid), methoxymethyl ether (MOM) (removed with acid) , methoxytrityl or [(4-methoxyphenyl)diphenylmethyl](MMT) (removed by acid and by hydrogenolysis), p-methoxybenzyl ether (PMB) (removed by acid, by hydrogenolysis or oxidation), methylthiomethyl ether (removed by acid ), pivaloyl (Piv) (removable with acid, base or reducing agents), tetrahydropyranyl (THP) (removable with acid), tetrahydrofuran (THF) (removable with acid), trityl (triphenylmethyl, Tr) (removable with acid and by hydrogenolysis), silyl ether (e.g. trimethylsilyl (TMS), tert-butyldimethylsilyl (TBDMS), triisopropylsilyloxymethyl (TOM) and triisopropylsilyl (TIPS) esters) (removed with acid or fluoride ion such as NaF, TBAF (tetra fluoride -n-butylammonium, HF-Py or HF-NEt3)), methyl ethers (removed by cleavage of trimethylsilyl iodide (TMSI) in dichloromethane, acetonitrile or chloroform or BBr3 in DCM), ethoxyethyl ethers (EE) (removed with 1H of hydrochloric acid).

Connections.

Provided herein are compounds that can be used as hemoglobin modulators. The compounds disclosed herein are believed to have an improved pharmacokinetic profile compared to known hemoglobin modulators while maintaining or improving efficacy. It is also believed that the compounds disclosed herein have an improved pharmacological safety profile compared to known hemoglobin modulators.

The present description provides a compound of formula I:

or an isotopically enriched analogue, stereoisomer, mixture of stereoisomers or prodrug thereof, or a pharmaceutically acceptable salt of each thereof, wherein X, Y, Z and R ¹ are as defined herein.

In some embodiments, X is CH or N. In some embodiments, X is CH. In some embodiments, X is N.

In some embodiments, Y is CH or N. In some embodiments, Y is CH. In some embodiments of the invention

- 9 045055 Tenia Y represents N.

In some embodiments, X is CH or N; and Y is CH. In some embodiments, X is CH; and Y is CH. In some embodiments, X is N; and Y is CH. In some embodiments, X is CH; and Y represents represents N. In some embodiments, X represents N; a Y represents N.

In some embodiments, Z is absent or CH2, O, or S. In some embodiments, Z is CH2, O, or S. In some embodiments, Z is O or S. In some embodiments, Z is absent or is CH2 or O. In some embodiments, Z is omitted. In some embodiments, Z is CH2. In some embodiments, Z is O. In some embodiments, Z is S.

In some embodiments, R ¹ is dihydroxy-(C ₁ - ₄ alkyl), -CH2CH2OCH3, -CH2CH2CN or monohydroxy-(C _1-4 alkyl),

In some embodiments, R ¹ is dihydroxy-(C ₁ _ ₄ alkyl), -CH2CH2CN or monohydroxy-(C _1-4 alkyl),

In some embodiments, R ¹ is monohydroxy-(C _1-4 alkyl), dihydroxy-(C ₁ _ ₄ alkyl), -CH2CH2OCH3, or -CH2CH2CN. In some embodiments, R ¹ is monohydroxy-(C ₁ - ₄ alkyl), dihydroxy-(C _{1 4} alkyl), or -CH2CH2CN.

In some embodiments of -CH2CH2OCH3, -CH2CH2CN or the invention, R ¹ is

-CH2OH,

-CH2CH2OH,

In some embodiments, -CH2CH2CN or

In some implementations

invention R ¹

of the invention R ¹ represents

-CH2OH,

- CH2CH2OH,

-CH2OH,

- CH2CH2OH,

- CH2CH2OCH3 or -CH2CH2CN. In some embodiments, R ¹ is -CH2OH, -CH2CH2OH, or -CH2CH2CN. In some embodiments, R ¹ is -CH2OH or -CH2CH2OH. In some embodiments, R ¹ is -CH2OH. In some embodiments, R ¹ is -CH2CH ₂ OH. In some embodiments, R ¹ is -CH2CH2OCH3. In some embodiments, R ¹ is -CH ₂ CH ₂ CN. In some embodiments, R ¹ is

In some embodiments, R ¹ is monohydroxy-(C1-4alkyl) or dihydroxy-(C1-4alkyl). In some embodiments, R ¹ is monohydroxy-(C1-4alkyl). In some embodiments, R ¹ is dihydroxy-(C2_4alkyl). In some embodiments, R ¹ is monohydroxy(C1-3alkyl) or dihydroxy-( _{C1_3alkyl} ). In some embodiments, R ¹ is monohydroxy-(C1_3alkyl). In some embodiments, R ¹ is dihydroxy-( _C2-3 alkyl). In some embodiments _, R ¹ is monohydroxy-(C _1-3 alkyl) or dihydroxy-(C _1-2 alkyl).

In some embodiments, R ¹ is hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, 1,2-dihydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl, or 2hydroxy-2-methylpropyl. In some embodiments, R ¹ is hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, 1,2-dihydroxyethyl, or 2-hydroxypropyl. In some embodiments, R ¹ is hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, or 2-hydroxypropyl.

In some embodiments, R ¹ is hydroxymethyl (ie -CH ₂ OH) or 2-hydroxyethyl (ie -CH ₂ CH ₂ OH). In some embodiments of the invention, R ¹

- 10 045055 is hydroxymethyl.

In some embodiments, R ¹ is 1-hydroxyethyl or 2-hydroxyethyl. In some embodiments, R ¹ is 1-hydroxyethyl. In some embodiments, R ¹ is

In some embodiments, R ¹ is

In some embodiments, R ¹ is 2-hydroxyethyl. In some embodiments, R ¹ is 1,2-dihydroxyethyl. In some embodiments, R ¹ is .OH\OH

In some embodiments, R ¹ is /OH

X ' ^7/ he

In some embodiments, R ¹ is 2-hydroxypropyl. In some embodiments, R ¹ is

In some embodiments, R ¹ is

In some embodiments, R ¹ is 3-hydroxypropyl.

In some embodiments, R ¹ is 2-hydroxy-2-methylpropyl.

In some embodiments, R ¹ is

or

R ^1c _R 1a %Y ^R,b Part where R ^1a represents hydrogen or methyl; R ^1b is hydrogen or methyl; and R ^1c is hydrogen or hydroxy.

Any combinations of X, Y, Z and R ¹ are included and provided in the present invention.

Some embodiments of the invention provide a compound of formula I:

or an isotopically enriched analogue, stereoisomer, mixture of stereoisomers or prodrug thereof, or a pharmaceutically acceptable salt of each, wherein:

X represents CH or N; Y represents CH or N; Z is absent or represents CH ₂ , O or S; and

R ¹ represents -CH2OH, -CH2CH2OH, -CH ₂ CH ₂ OCH ₃ , -CH2CH2CN or

- 11 045055

Some embodiments of the invention provide a compound of formula I:

I, or an isotopically enriched analogue, stereoisomer, mixture of stereoisomers or prodrug thereof, or a pharmaceutically acceptable salt of each, wherein:

X represents CH or N; Y represents CH or N; Z is absent or represents CH2 or O; and

R ¹ is -CH2OH, -CH2CH2OH, -CH2CH2OCH3, -CH2CH2CN or

In some embodiments, Y is CH; and Z is CH2.

Some embodiments of the invention provide a compound of Formula Ia:

or an isotopically enriched analogue, stereoisomer, mixture of stereoisomers or prodrug thereof, or a pharmaceutically acceptable salt of each, wherein:

X represents CH or N; and

R ¹ is -CH2OH, -CH2CH2OH, -CH2CH2OCH3, -CH2CH2CN or

In some embodiments, X is CH or N; Y represents CH; Z represents CH2, O or S; and R ¹ represents a monohydroxy-(C ₁ . ₄ alkyl) or dihydroxy-(C ₁ . ₄ alkyl) moiety as defined herein.

In some embodiments, X is CH or N; Y represents CH; Z represents CH2, O or S; and R ¹ is a monohydroxy-(C ₁ . ₄ alkyl) moiety as defined herein.

In some embodiments, X is N; Y represents CH; Z represents CH2, O or S; and R ¹ is a monohydroxy-(C ₁ . ₄ alkyl) moiety as defined herein.

In some embodiments, X is N; Y represents CH; Z represents O or S; and R ¹ is a monohydroxy-(C ₁ . ₄ alkyl) moiety as defined herein.

In some embodiments, X is N; Y represents CH; Z represents CH2, O or S; and R ¹ is -CH2OH or -CH2CH2OH. In some embodiments, X is N; Y represents CH; Z represents O or S; and R ¹ is -CH2OH or -CH2CH2OH. In some embodiments, X is N; Y represents CH; Z represents CH2; and R ¹ is -CH2OH or -CH2CH2OH. In some embodiments, X is N; Y represents CH; Z represents O; and R ¹ is -CH2OH or -CH2CH ₂ OH. In some embodiments, X is N; Y represents CH; Z represents S; and R ¹ is -CH ₂ OH or -CH ₂ CH ₂ OH.

- 12 045055

Some embodiments of the invention provide a compound of Formula Ib:

or an isotopically enriched analogue, stereoisomer, mixture of stereoisomers or prodrug thereof, or a pharmaceutically acceptable salt of each, wherein:

Z represents CH2, O or S; and

R ¹ represents a monohydroxy-(C ₁ . ₄ alkyl) moiety as defined herein.

In some embodiments, X is CH or N; Y represents CH; Z represents CH2, O or S; and R ¹ is a dihydroxy-(C2.4alkyl) moiety as defined herein. In some embodiments, X is CH; Y represents CH; Z represents CH2, O or S; and R ¹ is a dihydroxy-(C2.4alkyl) moiety as defined herein. In some embodiments, X is CH; Y represents CH; Z represents O or S; and R ¹ is a dihydroxy-(C2.4alkyl) moiety as defined herein. In some embodiments, X is CH; Y represents CH; Z represents CH2; and R ¹ is a dihydroxy-(C2.4alkyl) moiety as defined herein. In some embodiments, X is CH; Y represents CH; Z represents O; and R ¹ is a dihydroxy-(C2.4alkyl) moiety as defined herein. In some embodiments, X is CH; Y represents CH; Z represents S; and R ¹ is a dihydroxy-(C2.4alkyl) moiety as defined herein.

Some embodiments of the invention provide a compound of formula Ic:

or an isotopically enriched analogue, stereoisomer, mixture of stereoisomers or prodrug thereof, or a pharmaceutically acceptable salt of each, wherein:

Z represents CH2, O or S; and

R ¹ represents a dihydroxy(C^alkyl) moiety as defined herein.

- 13 045055

The present description provides a compound of the formula:

or an isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, or a pharmaceutically acceptable salt of each.

- 14 045055

The present description provides a compound of the formula:

or an isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, or a pharmaceutically acceptable salt of each.

- 15 045055

The present description provides a compound of the formula:

or an isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, or a pharmaceutically acceptable salt of each.

The present description provides a compound of the formula:

or an isotopically enriched analog, stereoisomer, mixture of stereoisomers, or prodrug thereof, or a pharmaceutically acceptable salt of each.

- 16 045055

The present description provides a compound of the formula:

or a pharmaceutically acceptable salt of each.

In some embodiments of the invention, the compound is:

or a pharmaceutically acceptable salt thereof.

In some embodiments of the invention, the compound is:

In some embodiments of the invention, the compound is:

or a pharmaceutically acceptable salt thereof.

In some embodiments of the invention, the compound is:

- 17 045055

In some embodiments of the invention, the compound is:

or a pharmaceutically acceptable salt thereof.

In some embodiments of the invention, the compound is:

In some embodiments of the invention, the compound is:

or a pharmaceutically acceptable salt thereof.

In some embodiments of the invention, the compound is:

In some embodiments of the invention, the compound is:

or a pharmaceutically acceptable salt thereof.

In some embodiments of the invention, the compound is:

- 18 045055

In some embodiments of the invention, the compound is:

or a pharmaceutically acceptable salt thereof.

In some embodiments of the invention, the compound is:

In some embodiments of the invention, the compound is:

or a pharmaceutically acceptable salt thereof.

In some embodiments of the invention, the compound is:

In some embodiments of the invention, the compound is:

or a pharmaceutically acceptable salt thereof.

In some embodiments of the invention, the compound is:

- 19 045055

In some embodiments of the invention, the compound is:

or a pharmaceutically acceptable salt thereof.

In some embodiments of the invention, the compound is:

In some embodiments of the invention, the compound is:

or a pharmaceutically acceptable salt thereof.

In some embodiments of the invention, the compound is:

In some embodiments of the invention, the compound is:

or a pharmaceutically acceptable salt thereof.

In some embodiments of the invention, the compound is:

- 20 045055

In some embodiments of the invention, the compound is:

or a pharmaceutically acceptable salt thereof.

In some embodiments of the invention, the compound is:

In some embodiments of the invention, the compound is:

or a pharmaceutically acceptable salt thereof.

In some embodiments of the invention, the compound is:

In some embodiments of the invention, the compound is:

or a pharmaceutically acceptable salt thereof.

In some embodiments of the invention, the compound is:

In some embodiments of the invention, the compound is:

- 21 045055 or a pharmaceutically acceptable salt thereof.

In some embodiments of the invention, the compound is:

In the present description, a compound selected from table is proposed. 1, or an isotopically enriched analogue, stereoisomer, mixture of stereoisomers, or prodrug thereof, or a pharmaceutically acceptable salt of each. In the present description, a compound selected from table is proposed. 1, or a pharmaceutically acceptable salt thereof. In the present description, a compound selected from table is proposed. 1.

In the present description, a compound selected from table is proposed. 2, or an isotopically enriched analogue, stereoisomer, mixture of stereoisomers, or prodrug thereof, or a pharmaceutically acceptable salt of each. In the present description, a compound selected from table is proposed. 2, or a pharmaceutically acceptable salt thereof. In this description, a compound selected from the table is proposed. 2.

The numbers of compounds and names of compounds according to the IUPAC nomenclature indicated in this description are summarized in table. 1 and 2.

Table 1

Connection number Structure IUPAC name 1 t o=< o ¹ \ / O --V \=O o (8)-2-hydroxy-6-(( 1 -(2-(2hydroxyethyl)nicotinoyl)piperidin-2- yl)methoxy)benzaldehyde 2 / o^° ^5 O ^Z \---to O (8)-2-hydroxy-6-(( 1 -(2-(2methoxyethyl)nicotinoyl)piper idin-2- yl)methoxy)benzaldehyde 3 °Ch/ ^e o (8)-3-(3-(2-((2-formyl-3hydroxyphenoxy)methyl)piperidine-1-carbonyl)pyridin-2yl)propanenitrile

- 22 045055

4 / ^T τ ο=ς o ο CV ^O ^ *—2 ^2 ⁴ --h }=O O (S)-2-hydroxy-6-(( 1-(2-(2- (pyrrolidine-1- yl)ethyl)nicotinoyl)piperidin2-yl)methoxy)benzaldehyde 5 o o=( 2^ ¹ o ⁷ )=o ^t ΞΕ (8)-2-hydroxy-6-((1-(2- (hydroxymethyl)benzoyl)piperidin-2-yl)methoxy)benzaldehyde 6 F I o=< o ,—ν^θт \ About A-Z -y )=O W (8)-2-hydroxy-6-((1 -(2-(2hydroxyethyl)benzoyl)piperidin-2-yl)methoxy)benzaldehyde 7 / ^T t ο=ς o /--h _c /° \~^ \ / O A-2 \^=o ( 8)-2-hydroxy-6-(( 1-(3-(2hydroxyethyl)pyrazine-2carbonyl)piperidin-2yl)methoxy)benzaldehyde

- 23 045055

8 O ° \ '—\ O / \ I O /=o ^t z/ (8)-2-hydroxy-6-((4-(2-(2hydroxyethyl)nicotinoyl)morph oline-3 - yl)methoxy)benzaldehyde 9 .-He Oh Oh (8)-2-hydroxy-6-(( 1 -(2-(2hydroxyethyl)nicotinoyl)pyrrolidin-2- yl)methoxy)benzaldehyde 10 (Enantiomer 2) / ¹ t o = ( o x \ / O \\ 5 = o 2-hydroxy-6-((4-(2-(2hydroxyethyl)nicotinoyl)thiome orpholine-3- yl)methoxy)benzaldehyde 10 (Enantiomer 1) ^o= \ '—\ o __/ \ I O )=o ^T I 2-hydroxy-6-((4-(2-(2hydroxyethyl)nicotinoyl)thiome orpholine-3- yl)methoxy)benzaldehyde AND O o = ( ¹ O ) = o ^t s: (U)-2-hydroxy-6-(( 1 -(2- (hydroxymethyl)nicotinoyl)pi peridin-2- yl)methoxy)benzaldehyde

-24045055

12 OH ⁰ C o x- (5)-2-hydroxy-6-((4-(2- (hydroxymethyl)nicotinoyl)mo rpholin-3- yl)methoxy)benzaldehyde 13 (Enantiomer 1) Is he X? ⁰ H O X: 2-hydroxy-6-((4-(2- (hydroxymethyl)nicotinoyl)thiomorpholine-3 - yl)methoxy)benzaldehyde 13 (Enantiomer 2) Is he X? ⁰ ^0 0 X: 2-hydroxy-6-((4-(2- (hydroxymethyl)nicotinoyl)thiomorpholine-3 - yl)methoxy)benzaldehyde 14 ci? ⁰ X 0 X: (8)-2-hydroxy-6-(( 1 -(2-(2methoxyethyl)benzoyl)piperidin-2-yl)methoxy)benzaldehyde 15 ci? ⁰ X 0 X (S)-3 -(2-(2-((2-formyl-3 - hydroxyphenoxy)methyl)piperi din-1- carbonyl)phenyl)propanenitrile

- 25 045055

16 τ ο=ς o χ. \ ABOUT \--у )=О (8)-2-hydroxy-6-(( 1 -(3 -(2hydroxyethyl)picolinoyl)piperidin-2- yl)methoxy)benzaldehyde 17 qXDO o \=o ^t A (8)-2-hydroxy-6-(( 1 -(2-(2- (pyrrolidine-1- yl)ethyl)benzoyl)piperidin-2yl)methoxy)benzaldehyde 18 he ⁰ H 0 όό (8)-2-hydroxy-6-(( 1 -(2-(3 hydroxypropyl)nicotinoyl)pi peridin-2- yl)methoxy)benzaldehyde 19 J - \ Ζ , , —_/ I o ¹ o /=o ^t X (8)-2-hydroxy-6-(( 1 -(3 -(2hydroxyethyl)pyrazine-2carbonyl)pyrrolidin-2yl)methoxy)benzaldehyde 20 o ^o= ( —\ O— O /=O ^t I (8)-2-hydroxy-6-((4-(2-(2methoxyethyl)nicotinoyl)morpholine-3-yl)methoxy)benzaldehyde

- 26 045055

21 z. z // ^O= \ '—\ O 0^0 (8)-2-hydroxy-6-((4-(3 -(2hydroxyethyl)pyrazine-2carbonyl)morpholin-3 yl)methoxy)benzaldehyde 22 o ^o= \ '—\ O fVo^O o /=o ^T I (8)-2-hydroxy-6-((4-(2-(2hydroxyethyl)benzoyl)morpholyl n-3-yl)methoxy)benzaldehyde 23 OH c^2° ⁰ % 0 00 (8)-2-hydroxy-6-((4-(2(hydroxymethyl)benzoyl)morpho-lin-3-yl)methoxy)benzaldehyde 24 o 0 όθ (8)-2-hydroxy-6-(( 1 -(2-(2methoxyethyl)nicotinoyl)pyrrolidine-2- yl)methoxy)benzaldehyde 25 Ό 0 00 (8)-2-hydroxy-6-(( 1 -(3 -(2methoxyethyl)pyrazine-2carbonyl)pyrrolidin-2yl)methoxy)benzaldehyde

- 27 045055

26 HE O O % (8)-2-hydroxy-6-(( 1-(2- (hydroxymethyl)benzoyl)pyrrole idine-2- yl)methoxy)benzaldehyde 27 / ^T t o o~\ -o ^z \^=o vy (8)-2-hydroxy-6-((4-(3 -(2methoxyethyl)pyrazine-2carbonyl)morpholin-3 yl)methoxy)benzaldehyde 28 ⁰ H O % (S)-3-(3-(3-((2-formyl-3hydroxyphenoxy)methyl)morphol in-4-carbonyl)pyridin-2yl)propanenitrile 29 o ^o= \ '—\ Z^ o — o /=o ¹ I (8)-2-hydroxy-6-((4-(2-(2methoxyethyl)benzoyl)morpholine -3-yl)methoxy)benzaldehyde thirty ll\Z ζ. Ά /=\ /~< J \\\ ΛΑ ° ^Ζ o )=o ¹ I (8)-3-(3-(2-((2-formyl-3hydroxyphenoxy)methyl)pyrrole idin-1-carbonyl)pyridin-2yl)propanenitrile

- 28 045055

31 / ^t t Kt _z /°A^°CHE ^z '—\ \7 (S)-3 -(2-(3 -((2-formyl-3 hydroxyphenoxy)methyl)morphol in-4- carbonyl)phenyl)propanenitrile 32 t ο=ς o Z C _z '—ξ )=O -TA ζ^__ζ (S)-3 -(3 -(3 -((2-formyl-3 hydroxyphenoxy)methyl)morphol in-4-carbonyl)pyrazin-2yl)propanenitrile 33 ^Z A / o=( Yq z^ ¹ fVo^o o )=o ^t I (8)-2-hydroxy-6-((4-(3 (hydroxymethyl)pyrazine-2carbonyl)morpholin-3 yl)methoxy)benzaldehyde 34 o t t o o ^z >°t / ⁼⁼ \ ° 2-(((2S)-l -(2-(1,2- dihydroxyethyl)benzoyl)piperidin-2-yl)methoxy)-6- hydroxybenzaldehyde 34 (Diastereomer 1) o t t o o ^z / ⁼⁼ \ ° 2-(((2S)-l -(2-(1,2- dihydroxyethyl)benzoyl)piperidin-2-yl)methoxy)-6- hydroxybenzaldehyde

-29045055

34 (Diastereomer 2) Oh >=/ Z--k сН—' o o t t ABOUT 2-(((2S)-l-(2-(l,2- dihydroxyethyl)benzoyl)piperidin-2-yl)methoxy)-6- hydroxybenzaldehyde 35 (Diastereomer 1) 5V? 1 ^he ° 9 9 os 2-(((3R)-4-(2-(l,2- dihydroxyethyl)benzoyl)thiomor folin-3-yl)methoxy)-6- hydroxybenzaldehyde 35 (Diastereomer 2) ι °н° Ϊ 8 00; 2-(((3 R)-4-(2-( 1,2- dihydroxyethyl)benzoyl)thiomor folin-3-yl)methoxy)-6- hydroxybenzaldehyde 36 O I t o o '—Z >°x lH/° X/\ 2-{[(2S)-l-[2-(l,2- dihydroxyethyl)pyr idin-3 carbonyl] piper idin-2yl] methoxy}-6hydroxybenzaldehyde 37 X ABOUT \-y ^=O ABOUT (K)-2-hydroxy-6-((4-(2-(2hydroxyethyl)nicotinoyl)morph oline-3 - yl)methoxy)benzaldehyde

- 30 045055

38 o ^o= \ '—\ O o )=o ¹ I (K)-2-hydroxy-6-((1 -(2-(2hydroxyethyl)nicotinoyl)piperidin-2yl)methoxy)benzaldehyde 39 l o o )=o ^t I (8)-2-hydroxy-6-((4-(2-(2- hydroxy-2- methylpropyl)nicotinoyl)morph oline-3 - yl)methoxy)benzaldehyde 40 m o =( °\>° 2-hydroxy-6-((4-(2- (hydroxymethyl)benzoyl)thiomor folin-3 - yl)methoxy)benzaldehyde 40 (Enantiomer 1) he θγ9θ ⁰ o όό 2-hydroxy-6-((4-(2- (hydroxymethyl)benzoyl)thiomor folin-3- yl)methoxy)benzaldehyde 40 (Enantiomer 2) he ⁰ X o сХ: 2-hydroxy-6-((4-(2- (hydroxymethyl)benzoyl)thiomor folin-3- yl)methoxy)benzaldehyde

- 31 045055

41 (Diastereomer 1) BUT ^2° ⁰ H O OS: 2-hydroxy-6-(((3 S)-4-(2-( 1 hydroxyethyl)nicotinoyl)morph oline-3 - yl)methoxy)benzaldehyde 41 (Diastereomer 2) but ^2 ⁰ X 0 00 2-hydroxy-6-(((3 S)-4-(2-( 1 hydroxyethyl)nicotinoyl)morph oline-3 - yl)methoxy)benzaldehyde 42 (Diastereomer 1) o ^o= \ '—\ O o /=o ^t x 2-hydroxy-6-(((3 S)-4-(2-(2hydroxypropyl)nicotinoyl)mo rfolin-3- yl)methoxy)benzaldehyde 42 (Diastereomer 2) / ¹ t o /°l_/ ^he O X \ O }—\ )=° /j 2-hydroxy-6-(((3S)-4-(2-(2hydroxypropyl)nicotinoyl)mo rfolin-3yl)methoxy)benzaldehyde 43 (Diastereomer 1) but ^2 ⁰ X o 00 2-hydroxy-6-(((3K)-4-(2-(1hydroxyethyl)nicotinoyl)thiome orpholin-3- yl)methoxy)benzaldehyde 43 (Diastereomer 2) But ° Ό O OO 2-hydroxy-6-(((3K)-4-(2-(1hydroxyethyl)nicotinoyl)thiome orpholin-3- yl)methoxy)benzaldehyde 44 (Diastereomer 1) / ^X X O=< O X \ O }—\ )= ^o 2-hydroxy-6-(((3K)-4-(2-(2hydroxypropyl)nicotinoyl)thiomorpholin-3yl)methoxy)benzaldehyde 44 (Diastereomer 2) o ο=ξ '—( z.—' ° 0^0 2-hydroxy-6-(((3K)-4-(2-(2hydroxypropyl)nicotinoyl)thiomorpholin-3yl)methoxy)benzaldehyde

- 32 045055

table 2

- 33 045055

- 34 045055

-35045055

In the present description, a compound selected from table is proposed. 6, or an isotopically enriched analogue, stereoisomer, mixture of stereoisomers, or prodrug, or a pharmaceutically acceptable salt of each thereof, excluding reference compounds A, B, and C. Provided herein is a compound selected from Table 6. 6, or a pharmaceutically acceptable salt thereof, excluding comparative compounds A, B and C. The present specification provides a compound selected from Table. 6, with the exception of comparative compounds A, B and C.

In the present description, a compound selected from table is proposed. 7, or an isotopically enriched analogue, stereoisomer, mixture of stereoisomers or prodrug, or a pharmaceutically acceptable salt of each thereof, excluding comparative compounds A and B. The present specification provides a compound selected from Table 7. 7, or a pharmaceutically acceptable salt thereof, excluding comparative compounds A and B. The present description provides a compound selected from table. 7, with the exception of comparative compounds A and B.

Methods of treatment and application

A therapy or treatment is an approach to obtain preferred or desired results, including results from clinical trials. Preferred or desired results of clinical trials may include one or more of the following characteristics: a) inhibition of a disease or condition (eg, reduction of one or more symptoms resulting from the disease or condition and/or reduction in the severity of the disease or condition); b) slowing or suppressing the development of one or more clinical symptoms associated with a disease or condition (eg, stabilizing a disease or condition, preventing or delaying the worsening or development of a disease or condition, and/or preventing or delaying the spread (eg, metastasis) of a disease or condition) ; and/or c) alleviation of the disease, that is, regression of clinical symptoms (for example, improvement of the disease, providing partial or complete remission of the disease or condition, enhancing the effect of another drug, slowing the progression of the disease, improving quality of life and/or increasing survival.

Prophylaxis or prevention means any treatment of a disease or condition that causes no clinical symptoms of the disease or condition to develop. In some embodiments, the compounds can be administered to a subject (including a human) who is at risk or has a genetic predisposition to a disease or condition.

- 36 045055

Subject refers to an animal, such as a mammal (including a human), that has been or will be the subject of treatment, observation, or experiment. The methods described herein may be useful in human and/or veterinary medicine. In some embodiments, the subject is a mammal. In one embodiment of the invention, the subject is a human.

The term "therapeutically effective amount" or effective amount of a compound as defined herein, or a pharmaceutically acceptable salt, tautomer, stereoisomer, mixture of stereoisomers, prodrug or deuterated analogue thereof, means an amount sufficient to effectively treat when administered to a subject to provide a therapeutic effect, such as relief of symptoms. or slowing the progression of the disease. For example, a therapeutically effective amount may be an amount sufficient to reduce a symptom of sickle cell disease. The therapeutically effective amount may vary depending on the subject and the disease or condition being treated, the weight and age of the subject, the severity of the disease or condition, and the route of administration, which can be readily determined by one skilled in the art.

The methods described herein can be applied to cell populations in vivo or ex vivo. In vivo means inside a living individual, such as inside an animal or a human being. In this context, the methods described herein can be used therapeutically for an individual. Ex vivo means outside of a living individual. Examples of ex vivo cell populations include in vitro cell cultures and biological samples, including fluid or tissue samples obtained from individuals. Such samples can be prepared by methods well known in the art. Examples of body fluid samples include blood, cerebrospinal fluid, urine, and saliva. In this context, the compounds and compositions described herein can be used for a variety of purposes, including therapeutic and experimental purposes. For example, the compounds and compositions described herein can be used ex vivo to determine the optimal schedule and/or dosage for administration of a compound of the present invention for a given indication, cell type, individual, and other parameters. Information obtained from such applications can be used experimentally or clinically to develop in vivo treatment protocols. Other ex vivo applications for which the compounds and compositions described herein may be suitable are described below or will be apparent to those skilled in the art. Selected compounds may be further characterized for dosage testing for safety or tolerability in humans or non-human subjects. Such properties can be examined using methods well known to those skilled in the art.

As used herein, the term hemoglobin refers to any hemoglobin protein, including normal hemoglobin (HbA) and abnormal hemoglobin such as sickle hemoglobin (HbS).

The term sickle cell disease refers to sickle hemoglobin (HbS)-mediated diseases that result from a single point mutation in hemoglobin (Hb). Sickle cell diseases include sickle cell disease (HbSS), hemoglobin S-C disease (HbSC), hemoglobin S-beta-plus-thalassemia (HbS/e+) and hemoglobin S-beta-null-thalassemia (HbS/e0).

Provided herein are methods of treating sickle cell disease (SCD). Sickle hemoglobin (HbS) contains a point mutation that replaces glutamic acid with valine, making sickle hemoglobin susceptible to polymerization under hypoxic conditions, giving sickle hemoglobin-containing red blood cells their characteristic sickle shape. Sickle cells are also stiffer than normal red blood cells, and their inherent lack of flexibility can lead to clogged blood vessels. It should be borne in mind that the approach to therapy is to maintain sickle hemoglobin in an oxygenated state, since under hypoxic conditions polymerization occurs only in an anoxic state.

In some embodiments, provided herein is a method of increasing the oxygen affinity of hemoglobin type S in a subject in need thereof, comprising administering to the subject a compound as defined herein, or an isotope-enriched analogue thereof, a stereoisomer, a mixture of stereoisomers, or a prodrug, or pharmaceutically an acceptable salt of each, or a pharmaceutical composition as defined herein. In some embodiments, provided herein is a method of increasing the oxygen affinity of hemoglobin type S in a subject in need thereof, comprising administering to the subject a compound as defined herein or a pharmaceutical composition as defined herein.

In some embodiments, provided herein is a method of treating a hemoglobin-mediated disorder in a subject in need thereof, comprising administering to the subject a compound as defined herein, or an isotope-enriched analogue thereof, a stereoisomer, a mixture of stereoisomers, or a prodrug. , or a pharmaceutically acceptable salt of each, or a pharmaceutical composition as defined herein. In some embodiments, provided herein is a method of treating a hemoglobin-mediated disorder in a subject in need thereof, comprising administering to the subject a compound as defined herein or a pharmaceutical composition as defined herein. In some embodiments, the disorder is a hemoglobinopathy.

In some embodiments, the hemoglobin is sickle hemoglobin.

In some embodiments, provided herein is a method of treating sickle cell disease in a subject in need thereof, comprising administering to the subject a compound as defined herein, or an isotope-enriched analogue, stereoisomer, mixture of stereoisomers, or prodrug, or a pharmaceutically acceptable salt thereof. each of them, or a pharmaceutical composition as defined herein. In some embodiments, provided herein is a method of treating sickle cell disease in a subject in need thereof, comprising administering to the subject a compound as defined herein or a pharmaceutical composition as defined herein.

Pharmaceutical compositions and methods of administration

The compounds provided herein are typically administered in the form of pharmaceutical compositions. Thus, the present specification also provides pharmaceutical compositions that contain one or more of the compounds described herein, or an isotopically enriched analogue of the compound, a stereoisomer thereof, a mixture of stereoisomers, or a prodrug, or a pharmaceutically acceptable salt of each thereof, and one or more pharmaceutically acceptable drug-carrying media selected from carriers, adjuvants and excipients. Suitable pharmaceutically acceptable drug-carrying media may include, for example, inert solid diluents and excipients, diluents including sterile aqueous solution and various organic solvents, permeation enhancers, solubilizers and adjuvants. Such compositions are prepared by methods well known in the pharmaceutical field. See, for example, Remington's Pharmaceutical Sciences, Mace Publishing Co., Philadelphia, Pa. 17th Ed. (1985); and Modern Pharmaceutics, Marcel Dekker, Inc. 3rd Ed. (G. S. Banker & C. T. Rhodes, Eds.).

The pharmaceutical compositions can be administered either once or multiple times. The pharmaceutical composition can be administered in a variety of ways, including, for example, rectal, buccal, intranasal and transdermal routes. In some embodiments, the pharmaceutical composition may be administered by intra-arterial injection, intravenous, intraperitoneal, parenteral, intramuscular, subcutaneous, oral, topical, or inhalation dosage form.

One route of administration is parenteral, for example by injection. Forms in which the pharmaceutical compositions described herein may be formulated for administration by injection include, for example, aqueous or oleaginous suspensions or emulsions with sesame oil, corn oil, cottonseed oil or peanut oil, as well as tinctures, mannitol, dextrose or sterile aqueous solution and similar pharmaceutical drug-carrying media.

Oral administration may be another route of administration for the compounds described herein. Administration can be carried out, for example, in capsules or enteric tablets. In the preparation of pharmaceutical compositions that include at least one compound described herein, or an isotopically enriched analogue, stereoisomer, mixture of stereoisomers or prodrug, or a pharmaceutically acceptable salt of each, the active ingredient is typically diluted with an excipient and/or enclosed in a carrier which is in the form of a capsule, sachet, paper or other container. When the excipient serves as a diluent, it may be in the form of a solid, semi-solid or liquid material that acts as a drug-carrying medium, carrier or vehicle for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, sachets, wafers, tinctures, suspensions, emulsions, solutions, syrups, aerosols (in solid or liquid medium), ointments containing, for example, up to 10 wt. .% active compound, soft and hard gelatin capsules, sterile injection solutions and powders in sterile packaging.

Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum arabic, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, sterile water, syrup and methylcellulose. Dosage forms may further include lubricants such as talc, magnesium stearate and mineral oil; wetting agents; emulsifiers and suspending agents; preservatives such as methyl and propyl hydroxybenzoates; sweeteners; and flavors.

Compositions that include at least one compound described herein, or an isotopically enriched analogue, stereoisomer, mixture of stereoisomers, or prodrug thereof, or

- 38 045055 a pharmaceutically acceptable salt of each of them can be prepared in such a way as to provide rapid, delayed or delayed release of the active ingredient after administration to a subject using procedures known in the art. Controlled-release drug delivery systems for oral administration include osmotic pump systems and dissolution systems containing polymer-coated reservoirs or polymer matrix drug dosage forms. Examples of controlled release systems are provided in US Pat. Nos. 3,845,770; 4,326,525; 4,902,514 and 5,616,345. Another dosage form for use in the methods disclosed herein uses transdermal delivery devices (patches). Such transdermal patches can be used to provide continuous or intermittent infusion of the compounds described herein in controlled amounts. The preparation and use of transdermal patches for the delivery of pharmaceutical agents is well known in the art. See, for example, US Patent Nos. 5,023,252, 4,992,445 and 5,001,139. Such patches can be prepared for continuous delivery, pulsatile delivery, or on-demand delivery of pharmaceutical agents.

To prepare solid compositions, such as tablets, the main active ingredient can be mixed with a pharmaceutical excipient to form a solid composition into a dosage form containing a homogeneous mixture of a compound described herein, or an isotope-enriched analogue thereof, a stereoisomer, a mixture of stereoisomers, or prodrugs thereof, or a pharmaceutically acceptable salt of each. When the compositions are referred to as homogeneous before dosage form, the active ingredient can be uniformly dispersed throughout the composition so that the composition can be easily divided into unit dosage forms such as tablets, pills and capsules.

Tablets or pills of the compounds described herein may be coated or otherwise prepared according to a prescription to provide a dosage form providing the benefit of extended release or for protection against acidic conditions of the stomach. For example, a tablet or pill may include an inner dosage component and an outer dosage component, the latter being in the form of a coating covering the former. The two components may be separated by an enteric layer, which serves to prevent disintegration in the stomach and allows the internal component to pass into the duodenum intact or delay the release of the internal component. A variety of materials can be used for such enteric layers or coatings, including a number of polymer acids and mixtures of polymer acids with materials such as shellac, cetyl alcohol and cellulose acetate.

Compositions for inhalation or insufflation may include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents or mixtures thereof, as well as powders. Liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as defined herein. In some embodiments, the compositions are administered by the oral or nasal respiratory route for local or systemic effect. In other embodiments, compositions in pharmaceutically acceptable solvents can be nebulized using inert gases. Nebulized solutions can be inhaled directly from the nebulizing device, or the nebulizing device can be attached to an oxygen tent with a breathing mask or to a positive intermittent pressure breathing apparatus. The compositions, in the form of a solution, suspension or powder, can be administered, preferably orally or nasally, from devices that deliver the dosage form appropriately.

Dosing.

The specific dosage of a compound herein for any particular subject will depend on a variety of factors, including the potency of the particular compound used, age, body weight, general health, sex, diet, time of administration, route of administration and rate of elimination, drug combination, and severity of the particular disease in the subject undergoing therapy. For example, the dosage may be expressed as the number of milligrams of a compound described herein per kilogram of subject body weight (mg/kg). Dosages from about 0.1 to 150 mg/kg are suitable. In some embodiments, suitable dosages range from 0.1 to 100 mg/kg. In other embodiments, a dosage of 0.5 to 60 mg/kg is suitable. Normalization according to the subject's body weight is especially useful when adjusting dosages between subjects of different sizes, for example, for use of a drug in both children and adults, or when converting the effective dosage in a non-human subject, such as a dog, to a dosage suitable for humans.

Synthesis of compounds.

The compounds can be prepared using the methods disclosed herein and routine modifications thereof, which will be apparent from the disclosure herein and methods well known in the art. In addition to the teachings set forth herein, standard and well known synthesis techniques can be used. The synthesis of typical compounds mentioned herein can be carried out as described in the following examples. When available, reagents can be purchased, for example, from Sigma Aldrich or other chemical suppliers.

General synthesis.

Exemplary embodiments of the compounds described herein can be synthesized using the general reaction schemes described below. It will be apparent from the description given in this section that the general designs can be modified by replacing the starting materials with other materials having similar structures to produce products that are correspondingly different. Syntheses are described below to provide numerous examples of how the starting materials can be varied to obtain the corresponding products. Given a target product for which groups of substituents have been defined, the required starting materials are usually determined through research. Starting materials are usually commercially available or synthesized using published procedures. For the synthesis of compounds that are embodiments described in the present invention, examination of the structure of the compound to be synthesized will ensure the identity of each group of substituents. The identity of the final product typically reveals the identity of the required starting materials through a simple research process, taking into account the examples provided herein. In general, the compounds described herein are generally stable and can be isolated at room temperature and standard pressure.

In some embodiments, a compound of Formula I can be synthesized through exemplary synthetic methods as shown in Schemes A and B.

In some embodiments, AR ² may be hydroxyl or chlorine; R ¹ may be monohydroxy-(C ₁ . ₄ alkyl), -CH ₂ CH ₂ OCH3, -CH ₂ CH ₂ CN or

and X, Y and Z are fragments as defined herein. As shown in Scheme A, compound A1 and compound A2 are first coupled using standard coupling conditions to produce compound A3, which is then reacted with 2,6-dihydroxybenzaldehyde A4 to produce a compound of formula I. In some embodiments, when R ¹ is monohydroxy -(C ₁ . ₄ alkyl) compounds of formula I, the hydroxy group of the R ¹ fragment in A1 includes a hydroxy protecting group known in the art; The protecting group can subsequently be removed after coupling A3 with A4 using standard procedures to prepare a compound of formula I.

Scheme A

In some embodiments of Scheme B, R ³ may be a C2.4 alkene; R ² may be hydroxyl or chlorine; R ¹ may be dihydroxy-( _C2.4 alkyl); Q is a halogen-containing compound; PG represents a hydroxy protecting group; and X, Y and Z are fragments as defined herein. As shown in Scheme B, compound B1 and compound B2 are first combined using standard coupling conditions to produce compound B3. Standard deprotection procedures provide compound B4, which is then reacted with 2,6-dihydroxybenzaldehyde A4 to produce compound B5. Introduction of an alkene (eg, via Stille-type coupling) produces compound B6, which can then be converted to the compound of formula I using dihydroxylation procedures known in the art.

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Scheme B

SNO he

Examples

The following examples are included to demonstrate specific embodiments of the invention. Those skilled in the art will appreciate that the methods disclosed in the examples that follow are methods that work well in the practice of the present invention, and thus can be considered to constitute specific modes of application thereof. However, in light of the present invention, those skilled in the art will appreciate that many changes may be made to specific embodiments of the invention and still obtain the same or similar result without departing from the spirit and scope of the invention.

Examples of synthesized compounds

Example 1. Synthesis of (S)-2-hydroxy-6-((1-(2-(2-hydroxyethyl)nicotinoyl)piperidin-2-yl)methoxy)benzaldehyde, compound 1.

Compound 1 was synthesized according to Scheme 1A.

Scheme 1A

Stages 1-4

1b (R = Η. X = Br)

1s (R = TBS _; X = Br)

Id (R = TBS _: X = COOMe)

1e(R TBS.X COOH)

Step 1. Synthesis of 2-(3-bromopyridin-2-yl)ethan-1-ol (1b).

A solution of ethyl 2-(3-bromopyridin-2yl)acetate (1a) (4 g, 16.39 mmol, 1 equiv) in tetrahydrofuran (THF) (40 ml) was placed into a 100 mL three-neck round-bottom flask. Diisobutylaluminum hydride (DIBAL-H) in THF (16 mL, 32.00 mmol,

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1.95 eq.) at -78°C. The resulting mixture was allowed to warm to room temperature and stirred for an additional 3 hours at 25°C. The reaction was then quenched by adding 50 ml of saturated NH4Cl. The resulting solution was extracted with 3x100 ml ethyl acetate and the organic layers were combined, washed with 2x100 ml brine. The separated organic layer was dried over Na ₂ SO ₄ , activated carbon, filtered and then concentrated. The residue was applied to a silica gel column and eluted with ethyl acetate/petroleum ether (1:3) to give the title compound. LCMS (liquid chromatography-mass spectrometry, LCMS) (ES, electrospray, ES) [M+1]+ m/z 202.0.

Step 2. Synthesis of 3-bromo-2-(2-((tert-butyldimethylsilyl)oxy)ethyl)pyridine (1c).

A solution of 2-(3-bromopyridin-2-yl)ethan-1-ol (1.9 g, 9.40 mmol, 1 eq.) in dimethylformamide (DMF) (20 ml) was placed in a 100 ml round bottom flask. 1H-imidazole (1.3 g, 18.81 mmol, 2 eq.), 4dimethylaminopyridine (DMAP) (0.1 g, 0.94 mmol, 0.1 eq.), tert-butyl(chloro)dimethylsilane (2 .8 g, 18.81 mmol, 2 eq.). The resulting solution was heated to 50°C and stirred for 2 hours. The reaction mixture was cooled and extracted with 2x50 ml ethyl acetate. The combined organic layers were washed with 2x50 ml brine, dried over Na2SO4, filtered and concentrated. The residue was applied to a silica gel column and eluted with ethyl acetate/petroleum ether (1:3) to give the title compound. LCMS (ES) [M+1]+ m/z 316.1.

Alternatively, a protecting group such as tert-butyldiphenylsilyl (TBDPS) can be used instead of tert-butyl(chloro)dimethylsilyl (TBS). Under typical conditions, imidazole (1.5 to 4 eq) and tert-butyl(chloro)diphenylsilane (TBDPSCl (about 1 eq)) were added to a solution of alcohol 1b (1 eq) in DCM (3 to 15 vol). . The reaction mixture was stirred at room temperature for a period of time from 1 to 48 hours. After normal processing and purification this gave the product (1c2). The tert-butyldiphenylsilyl group can be removed using tetrabutylammonium fluoride (TBAF; 1-3 eq.) according to typical literature conditions. Compounds 8 and 12 can be synthesized using tert-butyldiphenylsilyl as a protecting group.

Step 3. Synthesis of methyl 2-(2-((tert-butyldimethylsilyl)oxy)ethyl)nicotinate (1d).

A solution of 3-bromo-2-[2-[(tertbutyldimethylsilyl)oxy]ethyl]pyridine (2.0 g, 6.32 mmol, 1 equiv.) in methanol (MeOH, 100 ml) was placed in a 250 ml sealed tube. triethylamine (TEA 1.3 g, 12.65 mmol, 2 eq.) and [1,1'-bis-(diphenylphosphino)ferrocene]dichloropalladium(P) (Pd(dppf)Cl ₂ , 0.5 g, 0. 63 mmol, 0.1 eq.). The resulting solution was stirred for 16 hours at 100°C in a CO atmosphere (10 atm). After cooling to room temperature, the reaction mixture was filtered and the filtrate was concentrated. The resulting residue was applied to a silica gel column and eluted with ethyl acetate/petroleum ether (1:3) to obtain the title compound. LCMS (ES) [M+1]+ m/z 296.2.

Alternative synthesis option: scheme 1B.

Scheme 1B

Alternatively, phenyl formate can be used to replace CO gas as the carbonyl source in the presence of triethylamine (2 eq), catalytic amounts of palladium acetate (e.g. 0.02 eq) and tri-tert-butylphosphonium tetrafluoroborate (e.g. 0.08 eq.) to convert bromide 1c2 to carboxylate 1d2 in acetonitrile (3 to 10 volumes) by heating (80°C) for 2 to 48 hours, and then directly to carboxylic acid 1e2 by hydrolysis of the ester in basic aqueous conditions (2-8 equiv. K ₂ CO ₃ in water from 3 to 10 volumes; at a temperature of 50 to 80 ° C for 48 hours).

Step 4. Synthesis of 2-(2-((tert-butyldimethylsilyl)oxy)ethyl)nicotinic acid (1e).

A solution of methyl 2-[2-[(tert-butyldimethylsilyl)oxy]ethyl]pyridine-3-carboxylate (1.7 g, 5.75 mmol, 1 eq.) in MeOH (20 mL) was placed in a 100 mL round bottom flask. a solution of LiOH (275.6 mg, 11.51 mmol, 2 eq.) in H2O (5 ml). The resulting solution was stirred for 4 hours at 25°C. Water (10 ml) was added to the reaction mixture, and the crude product as a precipitate was collected by filtration. The crude product was then purified by Flash-Prep-HPLC under the following conditions (IntelFlash-1): column, C18 silica gel; mobile phase, H ₂ O:acetonitrile (ACN)=10:l, increasing to H ₂ O:acetonitrile=3:1 over 10 minutes to obtain the title compound. LCMS (ES) [M+1]+ m/z 282.1.

Step 5. Synthesis of (S)-(2-(2-((tert-butyldimethylsilyl)oxy)ethyl)pyridin-3-yl)(2-(hydroxymethyl)piperidin-1-yl)methanone (1f).

2-[2-[(tert-Butyldimethylsilyl)oxy]ethyl]pyridine-3-carboxylic acid (1.4 g, 4.97 mmol, 1 eq.), [(2S)piperidine-2- yl]methanol (0.9 g, 7.46 mmol, 1.5 eq.), N,N-diisopropylethylamine (DIEA, 1.3 g, 9.95

- 42 045055 mmol, 2 eq.), 1-[bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide hexafluorophosphate (HATU, 2.8 g, 7.46 mmol, 1.5 eq.) and 30 ml dichloromethane (DCM). The resulting reaction mixture was stirred for 2 hours at 25°C and then diluted with 60 ml of H2O. The organic phase was extracted with 3x50 ml ethyl acetate. The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated. The residue was applied to a silica gel column and eluted with ethyl acetate/petroleum ether (1:3) to give the title compound. LCMS (ES) [M+1]+ m/z 379.2.

Steps 6 and 7. Synthesis of (S)-2-hydroxy-6-((1-(2-(2-hydroxyethyl)nicotinoyl)piperidin-2yl)methoxy)benzaldehyde (compound 1).

A solution of [(2S)-1-(2-[2-[(tert-butyldimethylsilyl)oxy]ethyl]pyridin-3-carbonyl)piperidin-2-yl]methanol (900 mg, 2. 38 mmol, 1 eq.) in DCM (8 ml), 2,6dihydroxybenzaldehyde (656.7 mg, 4.75 mmol, 2 eq.) and triphenylphosphine (PPh ₃ , 1247.0 mg, 4.75 mmol, 2 eq. .). Thereafter, a solution of di-tert-butyl azodicarboxylate (DBAD, 1094.8 mg, 4.75 mmol, 2 eq.) in DCM (2 ml) was added dropwise with stirring at 0°C. The resulting solution was stirred for 2 hours at 25°C. The reaction mixture was concentrated and dissolved in 20 ml THF. Tetrabutylammonium fluoride (TBAF, 1243.1 mg, 4.75 mmol, 2 eq.) was added to the mixture. The resulting mixture was allowed to stir for 2 hours at 25°C. The reaction mixture was concentrated to obtain the crude product, which was purified by preparative-HPLC under the following conditions (Prep-HPLC-006): column, XBridge Prep C18 OBD column, 19mm x 150mm 5 µm; mobile phase, water (10 mmol/l NH ₄ HCO ₃ + 0.1% NH3H2O) and acetonitrile (14% to 35% phase B over 8 minutes, 95% retention within 1 minute, reduction to 14% within 1 minute, holding 14% for 1 minute); detector, UV 254 nm. This resulted in the compound specified in the header being obtained. 1HTEM NMR (300 MHz, 353K, dimethyl sulfoxide (DMSO) _-d6 ): δ 11.36 (s, 1H), 10.25 (s, 1H), 8.51 (dd, J=4.8, 1, 8 Hz, 1H), 7.51-7.23 (m, 3H), 6.7-6.5 (m, 2H), 5.15 (s, 1H), 4.59-3.98 (m , 3H), 3.78 (broad s, 2H), 3.17-2.86 (m, 4H), 1.83-1.37 (m, 6H). LCMS (ES) [M+1]+ m/z 385.2.

Example 2. (S)-2-Hydroxy-6-((1-(2-(2-methoxyethyl)nicotinoyl)piperidin-2-yl)methoxy)benzaldehyde, compound 2.

Compound 2 was synthesized according to Scheme 2.

Scheme 2

Step 1. Synthesis of methyl 2-ethenylpyridine-3-carboxylate (2b).

A mixture of methyl 2-chloropyridine-3-carboxylate (3 g, 17.48 mmol, 1.00 eq), dioxane (40 ml), water (4 ml), 2-ethenyl-4 was placed in a 100 ml round bottom flask. ,4,5,5-tetramethyl-1,3,2-dioxaborolane (5.39 g, 34.99 mmol, 2.00 eq.), Cs ₂ CO ₃ (11.40 g, 34.99 mmol, 2 .00 eq.) and tetrakis(triphenylphosphine)palladium(0) (Pd(PPh ₃ ) ₄ , 2.02 g, 1.75 mmol, 0.10 eq.). The resulting solution was stirred for 2 hours at 100°C under N2 atmosphere. The reaction mixture was cooled, filtered and concentrated in vacuo. The resulting residue was purified on a silica gel column, eluting with ethyl acetate/petroleum ether (1/2) to give 2-methyl-2-ethenylpyridine-3-carboxylate. LCMS (ES) [M+1]+ m/z: 164.1.

Step 2. Synthesis of methyl 2-(2-methoxyethyl)pyridine-3-carboxylate (2c).

A solution of methyl 2-ethenylpyridine-3-carboxylate (1.5 g, 9.19 mmol, 1.00 eq), methanol (20 ml) and aqueous hydrogen chloride (36%, 2 ml) was placed in a 100 ml round bottom flask. ). The resulting solution was stirred for 48 hours at 60°C. The mixture was cooled and then concentrated in vacuo to obtain methyl 2-(2-methoxyethyl)pyridine-3-carboxylate. LCMS (ES) [M+1]+ m/z: 196.1.

Step 3. Synthesis of 2-(2-methoxyethyl)pyridine-3-carboxylic acid (2d).

A solution of methyl 2-(2-methoxyethyl)pyridine-3 was placed in a 100 ml round bottom flask

- 43 045055 carboxylate (2.50 g, 12.81 mmol, 1.00 eq.), methanol (30 ml), H ₂ O (6 ml) and NaOH (2.56 g, 64.00 mmol, 5. 00 eq.). The resulting solution was stirred for 2 hours at 50°C. The reaction mixture was cooled and the pH was adjusted to 6 by adding aqueous hydrogen chloride (2 M). The mixture was extracted with 3x50 ml DCM/MeOH (10/1). The combined organic layers were washed with 1x100 ml brine, dried over anhydrous sodium sulfate and concentrated in vacuo to give (crude) 2-(2methoxyethyl)pyridine-3-carboxylic acid. LCMS (ES) [M+1]+ m/z: 182.1.

Step 4. Synthesis of [(2S)-1-[[2-(2-methoxyethyl)pyridin-3-yl]carbonyl]piperidin-2-yl]methanol (2e).

A solution of 2-(2-methoxyethyl)pyridine-3-carboxylic acid (600 mg, 3.31 mmol, 1.00 equiv.), dichloromethane (30 mL), (2S)-piperidine-2 was placed in a 100 mL round bottom flask. -ylmethanol (762 mg, 6.62 mmol, 2.00 eq.), N,N-diisopropylethylamine (855 mg, 6.62 mmol, 2.00 eq.) and 1-[bis(dimethylamino)methylene]-1H -1,2,3-triazolo[4,5-b]pyridinium-3-oxide-hexafluorophosphate (1.89 g, 4.97 mmol, 1.50 eq.). The resulting solution was stirred for 1 hour at room temperature. The crude reaction mixture was filtered and concentrated. The resulting residue was purified by reverse preparative HPLC (Prep-d8, 20-45 µM, 120 g, Tianjin Bonna-Agela Technologies; gradient elution from 15% CH3CN in water to 40% CH3CN in water over a 12-minute period when both solvent contained 0.1% ammonia) to obtain [(2S)-1-[[2-(2-methoxyethyl)pyridin-3yl]carbonyl]piperidin-2-yl]methanol. LCMS (ES) [M+1]+ m/z: 279.1.

Step 5. (S)-2-G hydroxy-6-(( 1 -(2-(2-methoxyethyl)nicotinoyl)piperidin-2-yl)methoxy)benzaldehyde (compound 2).

A solution of [(2S)-1-[[2-(2methoxyethyl)pyridin-3-yl]carbonyl]piperidin-2-yl]methanol (265 mg, 0.95 mmol, 1.00 eq.), dichloromethane (10 ml), 2,6-dihydroxybenzaldehyde (263 mg, 1.90 mmol, 2.00 eq.) and PPh ₃ (499 mg, 1.90 mmol, 2.00 eq.). A solution of dibenzyl azodicarboxylate (DBAD) (438 mg, 1.90 mmol, 2.00 eq) in DCM (5 mL) was added under nitrogen at 0°C. The resulting solution was stirred for 2 hours at room temperature. The resulting mixture was concentrated under vacuum. The residue was purified on a silica gel column, eluting with ethyl acetate/petroleum ether (1/1). The crude reaction mixture was filtered and subjected to reverse preparative HPLC (Prep-d8, 5 mM XBridge column, 19x150 mm, water; gradient elution from 22% _CH3CN in water to 42% CH3CN in water over a 6-minute period when both solvents contained 0.1% trifluoroacetic acid (TFA)) to produce 2hydroxy-6-[(1-[hydroxy[2-(2-methoxyethyl)piperidin-3-yl]methyl]piperidin-2)yl)methoxy]cyclohexane-1 -carbaldehyde. 1HTEM NMR (300 MHz, 353 K, DMSOC) δ 11.64 (s, 1H), 10.28 (s, 1H), 8.56 (dd, J= 5.1 Hz, 1.8 Hz, 1H) , 7.78-7.59 (m, 1H), 7.52 (t, J= 8.1 Hz, 1H), 7.39-7.21 (m, 1H), 6.72 (s, 1H ), 6.55 (d, J= 8.4 Hz, 1H), 5.19 (s, 1H), 4.33-4.21 (m, 3H), 3.83-3.57 (m, 2H), 3.30-3.08 (m, 4H), 3.04-2.84 (m, 2H), 2.01-1.82 (m, 1H), 1.82-1.55 ( m, 4H), 1.55-1.28 (m, 1H). LCMS (ES) [M+1]+m/z: 399.1.

Example 3. (S)-3 -(3 -(2-((2-Formyl-3-hydroxyphenoxy)methyl)piperidine-1-carbonyl)pyridin-2yl)propanenitrile, compound 3.

Compound 3 was synthesized according to Scheme 3.

Scheme 3

For ZZ Zs

Step 1. Synthesis of methyl 2-formyl nicotinate (3b).

A solution of methyl 2-methylpyridine-3-carboxylate (5 g, 33.08 mmol, 1 eq.) in dioxane (50 ml), (oxo-lambda4-selanylidene)oxidane (selenium dioxide) was placed in a 100 ml round bottom flask. 5.5 g, 49.61 mmol, 1.5 eq.). After stirring for 16 hours at 110°C, the reaction mixture was cooled to room temperature, concentrated and diluted with 100 ml of H2O. The mixture was then extracted

4x100 ml ethyl acetate and organic layers were combined. The organic layers were washed with 200 ml brine, dried over Na ₂ SO ₄ and concentrated. The resulting residue was applied to a silica gel column and eluted with ethyl acetate/petroleum ether (1:5) to obtain the title compound. LCMS (ES) [M+1]+ m/z 166.0.

Step 2. Synthesis of methyl-(E)-2-(2-cyanovinyl)nicotinate (3c).

A solution of methyl 2-formylpyridine-3-carboxylate (2.5 g, 15.14 mmol, 1 eq) in THF (30 mL) was placed in a 100 mL round bottom flask. After this, diethyl(cyanomethyl)phosphonate (3.2 g, 18.17 mmol, 1.2 eq.) was added at 0°C and (tert-butoxy)potassium (2.5 g, 22.0 eq.) was added in portions at 0°C. 71 mmol, 1.5 eq.). The resulting mixture was stirred for 16 hours at room temperature. Solids were filtered off.

The filtrate was diluted with 100 ml H2O and extracted with 2x80 ml ethyl acetate. The combined organic layers were washed with 2x100 ml brine, dried over Na2SO4, filtered and concentrated. The resulting residue was applied to a silica gel column and eluted with ethyl acetate/petroleum ether (1:5) to obtain the title compound. LCMS (ES) [M+1] ⁺ m/z 189.1.

Step 3. Synthesis of (E)-2-(2-cyanovinyl)nicotinic acid (3d).

A solution of methyl 2-[(1E)-2-cyanoeth-1-en-1yl]pyridin-3-carboxylate (1.4 g, 7.44 mmol, 1 eq.) in MeOH was placed in a 100 mL round bottom flask. 20 ml), a solution of NaOH (0.6 g, 14.88 mmol, 2 eq.) in H2O (4 ml). After stirring for 2 hours at room temperature, the reaction mixture was diluted with 10 ml H2O, the pH was adjusted to 6-7 with HCl (2 mol/l) and then concentrated. The crude product was purified by flash preparative-HPLC under the following conditions (IntelFlash-1): column, silica gel C18; mobile phase, H ₂ O:acetonitrile=10:1, increasing to H ₂ O:acetonitrile=1:1 over 10 minutes. This resulted in the compound specified in the header being obtained. LCMS (ES) [M+1] ⁺ m/z 175.0.

Step 4: Synthesis of 2-(2-cyanoethyl)nicotinic acid (3e).

A solution of 2-[(1E)-2-cyanoeth-1-en-1-yl]pyridin-3carboxylic acid (600 mg, 3.45 mmol, 1 equiv.) in MeOH (20 ml) was placed in a 100 ml round bottom flask. , palladium on carbon (Pd/C, 120 mg, 1.13 mmol, 0.33 eq.). The resulting solution was stirred for 16 hours at room temperature under H2 atmosphere (20 atm). Solids were filtered off. The filtrate was concentrated to give the crude product. LCMS (ES) [M+1] ⁺ m/z 177.1.

Step 5. Synthesis of (S)-3-(3-(2-(hydroxymethyl)piperidin-1-carbonyl)pyridin-2-yl)propanenitrile (3f).

A solution of 2-[(1E)-2-cyanoeth-1-en-1-yl]pyridin-3carboxylic acid (550 mg, 3.16 mmol, 1 eq.) in DMF (6 ml) was placed in a 50 ml round bottom flask. , [(2S)-piperidin-2-yl]methanol (545.6 mg, 4.74 mmol, 1.5 eq.), N,N-diisopropylethylamine (816.3 mg, 6.32 mmol, 2 eq. ), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide-hexafluorophosphate (1801.2 mg, 4.74 mmol, 1.5 equiv .). The resulting solution was stirred for 4 hours at room temperature. The mixture was diluted with 40 ml H2O and extracted with 3x30 ml ethyl acetate. The combined organic layers were washed with 30x30 ml brine, dried over Na2SO4, filtered and concentrated. The resulting residue was purified by flash preparative HPLC under the following conditions (IntelFlash-1): column, silica gel C18; mobile phase, H ₂ O:acetonitrile=10:1, increasing to H ₂ O:acetonitrile=1:1 over 10 minutes. This resulted in the compound specified in the header being obtained. LCMS (ES) [M+1] ⁺ m/z 274.1.

Step 6. Synthesis of (S)-3-(3-(2-((2-formyl-3-hydroxyphenoxy)methyl)piperidine-1carbonyl)pyridin-2-yl)propanenitrile (compound 3).

A solution of 3-[3-[(2S)-2-(hydroxymethyl)piperidine1-carbonyl]pyridin-2-yl]propanenitrile (200 mg, 0.73 mmol, 1 eq.) in DCM was placed in a 50 mL round bottom flask ( 3 ml), 2,6dihydroxybenzaldehyde (202.1 mg, 1.46 mmol, 2 eq.), PPh ₃ (383.8 mg, 1.46 mmol, 2 eq.). Di-tert-butyl azodicarboxylate (337.0 mg, 1.46 mmol, 2 eq.) was then added at 0°C. The resulting mixture was stirred for 2 hours at room temperature and then concentrated in vacuo; the residue was diluted with 5 ml of acetonitrile and filtered. The crude product was purified by preparative-HPLC under the following conditions (Prep-HPLC-007): column, SunFire Prep C18 OBD column, 150 mm 5 µm 10 nm; mobile phase, water (0.1% formic acid) and MeOH (40% to 55% phase B in 7 minutes, holding 95% in 1 minute, reducing to 40% in 1 minute, holding 40% in 1 minute); detector, UV. This resulted in the compound specified in the header being obtained. 1HTEM NMR (300 MHz, 353K, DMSO- _d6 ): δ 11.60 (broad s, 1H), 10.27 (broad s, 1H), 8.60 (dd, J= 4.8, 1.8 Hz, 1H), 7.61 (m, 2H), 7.52 (t, J= 8.4 Hz, 1H), 7.34 (dd, J= 7.6, 4.8 Hz, 1H), 6.72 (s, 1H), 6.55 (d, J= 8.4 Hz, 1H), 5.16 (s, 1H), 4.49 (wide s, 1H), 4.32 (dd, J= 10.3, 6.2 Hz, 1H), 3.21-2.89 (m, 6H), 1.911.46 (m, 6H). LCMS (ES) [M+1] ⁺ m/z 394.1.

- 45 045055

Example 4. (S)-2-r hydroxy-6-(( 1 -(2-(2-(pyrrolidin-1 -yl)ethyl)nicotinoyl)piperidin-2yl)methoxy)benzaldehyde, compound 4.

Compound 4 was synthesized according to Scheme 4.

Scheme 4

Step 1. Synthesis of methyl 2-ethenylpyridine-3-carboxylate (4b).

A mixture of methyl 2-chloropyridine-3-carboxylate (3 g, 17.48 mmol, 1.00 eq), dioxane (40 ml), water (4 ml), 2-ethenyl-4 was placed in a 100 ml round bottom flask. ,4,5,5-tetramethyl-1,3,2-dioxaborolane (5.39 g, 34.99 mmol, 2.00 eq.), CS2CO3 (11.40 g, 34.99 mmol, 2.00 eq. .) and Pd(PPh ₃ ^ (2.02 g, 1.75 mmol, 0.10 eq.). The resulting solution was stirred for 2 hours at 100°C under N2 atmosphere. The reaction mixture was cooled, filtered and concentrated in vacuum The resulting residue was purified on a silica gel column, eluting with ethyl acetate/petroleum ether (1/2) to give 2-methyl-2-ethenylpyridine-3-carboxylate. LCMS (ES) [M+1]+ m/z: 164.1 .

Step 2. Synthesis of 2-ethenylpyridine-3-carboxylic acid (4c).

A solution of methyl 2-ethenylpyridine-3-carboxylate (3.0 g, 18.39 mmol, 1 equiv.), MeOH (50 ml), H ₂ O (5 ml) and NaOH (3) was placed in a 100 ml round bottom flask. .7 g, 91.93 mmol, 5.0 eq.). After stirring for 2 hours at 50°C, the reaction mixture was cooled and the pH was adjusted to 5 by adding aqueous HCl (2 M). The resulting mixture was concentrated and diluted with 100 ml DCM. Solids were filtered off. The mixture was concentrated. 2-ethenylpyridine-3-carboxylic acid was obtained. LCMS (ES) [M+1]+ m/z: 150.1.

Step 3. Synthesis of tert-butyl-(2S)-2-[[(2,2-dimethyl-4-oxo-2,4-dihydro-1,3-benzodioxin-5yl)oxy]methyl]piperidine-1-carboxylate (4f).

Compound 4d can be synthesized by methods known in the art.

A solution of 5-hydroxy-2,2-dimethyl-2,4-dihydro1,3-benzodioxin-4-one (4d, 10.0 g, 51.50 mmol, 1 eq.), THF was placed in a 1000 ml round bottom flask. (300 ml), tert-butyl-(2S)-2(hydroxymethyl)piperidine-1-carboxylate (22.2 g, 103.12 mmol, 2.00 eq.) and PPh ₃ (40.5 g, 154, 49 mmol, 3 eq.). A solution of diisopropyl azodicarboxylate (DIAD, 31.2 g, 154.49 mmol, 3 eq.) in THF (30 mL) was added dropwise at 0° C. under a nitrogen atmosphere. The resulting solution was stirred for 4 hours at room temperature. The resulting mixture was concentrated. The residue was purified on a silica gel column, eluting with ethyl acetate/petroleum ether (1/3). Tert-butyl-(28)-2-[[(2,2dimethyl-4-oxo-2,4-dihydro-1,3-benzodioxin-5-yl)oxy]methyl]piperidine-1-carboxylate was obtained. LCMS (ES) [M+1]+ m/z: 392.2.

Step 4. Synthesis of 2,2-dimethyl-5-[[(2S)-piperidin-2-yl]methoxy]-2,4-dihydro-1,3-benzodioxin-4one (4f).

A solution of tert-butyl-(2S)-2-[[(2,2-dimethyl-4-oxo2,4-dihydro-1,3-benzodioxin-5-yl)oxy]methyl]piperidine was placed in a 50 ml round bottom flask -1-carboxylate (2.0 g, 5.10 mmol, 1 eq.), DCM (15 ml) and HCl/dioxane (4 M, 5 ml). The resulting solution was stirred for 1 hour at room temperature. The resulting mixture was concentrated. 2,2-Dimethyl-5-[[(2S)-piperidin-2yl]methoxy]-2,4-dihydro-1,3-benzodioxin-4-one was obtained. LCMS (ES) [M+1]+ m/z: 292.2.

Step 5. Synthesis of 5-[[(2S)-1-(2-ethenylpyridin-3-carbonyl)piperidin-2-yl]methoxy]-2,2-dimethyl2,4-dihydro-1,3-benzodioxin-4- she (4g).

A solution of 2,2-dimethyl-5-[[(2S)-piperidin-2yl]methoxy]-2,4-dihydro-1,3-benzodioxin-4-one hydrochloride (1.0 g) was placed in a 100 ml round bottom flask , 3.05 mmol, 1 eq.), DCM (50 ml, 786.50 mmol, 257.82 eq.), 2-ethenylpyridine-3-carboxylic acid (910.0 mg, 6.10 mmol, 2.00 eq.), N,N-diisopropylethylamine (2.0 g, 15.25 mmol, 5 eq.) and 1-[bis-(dimethylamino)methylene]-1H-1,2,3triazolo[4,5-b] pyridinium 3-oxide hexafluorophosphate (2.3 g, 6.10 mmol, 2 eq.) at 0°C. After stirring for 2 hours at room temperature, the reaction mixture was diluted with 50 ml DCM and washed with 3x50 ml brine. The organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified on a silica gel column, eluting with ethyl acetate/petroleum ether (1/1). 5-[[(2S)-1-(2-ethenylpyridin-3-carbonyl)piperidin-2-yl]methoxy]-2,2-dimethyl-2,4dihydro-1,3-benzodioxin-4-one was obtained. LCMS (ES) [M+1] ⁺ m/z: 423.2.

Step 6. Synthesis of 2,2-dimethyl-5-[[(2S)-1-[2-[2-(pyrrolidin-1-yl)ethyl]pyridin-3carbonyl]piperidin-2-yl]methoxy]-2, 4-dihydro-1,3-benzodioxin-4-one (4h).

A solution of 5-[[(2S)-1-(2-ethenylpyridin-3carbonyl)piperidin-2-yl]methoxy]-2,2-dimethyl-2,4-dihydro-1,3- was placed in a 50 ml round bottom flask. benzodioxin-4-one (650 mg, 1.54 mmol, 1 eq.), ethanol (20 ml), pyrrolidine (218.8 mg, 3.08 mmol, 2.00 eq.) and triethylamine (311.4 mg , 3.08 mmol, 2 equiv.). The reaction mixture was stirred for 16 hours at 85°C, cooled and concentrated in vacuo. The resulting residue was purified on a silica gel column, eluting with a dichloromethane/methanol mixture (10/1). 2,2-Dimethyl-5-[[(2S)-1-[2-[2-(pyrrolidin-1-yl)ethyl]pyridin-3carbonyl]piperidin-2-yl]methoxy]-2,4-dihydro was obtained -1,3-benzodioxin-4-one. LCMS (ES) [M+1] ⁺ m/z: 494.3.

Step 7. Synthesis of 2-(hydroxymethyl)-3-[[(2S)-1-[2-[2-(pyrrolidin-1-yl)ethyl]pyridin-3carbonyl]piperidin-2-yl]methoxy]phenol (4i ).

A solution of 2,2-dimethyl-5-[[(2S)-1-[2-[2(pyrrolidin-1-yl)ethyl]pyridin-3-carbonyl]piperidin-2-yl was placed in a 50 ml three-neck round bottom flask ]methoxy]-2,4-dihydro-1,3-benzodioxin-4one (500 mg, 1.01 mmol, 1 eq.) and THF (10 ml). Lithium aluminum hydride in THF solution (LiAlH ₄ THF solution, 2.03 mL, 1 M, 2.03 mmol, 2 eq.) was added dropwise to the solution at -78° C. under a nitrogen atmosphere. The resulting mixture was stirred at -78°C for 1 hour. The reaction mixture was heated to 0°C, and then 0.07 ml of H2O, 0.07 ml of 15% aqueous NaOH and 0.21 ml of H2O were added dropwise to the mixture. The mixture was warmed to room temperature and stirred for 30 minutes. Solids were filtered off. The filtrate was concentrated. 2-(hydroxymethyl)-3-[[(2S)-1-[2-[2-(pyrrolidin-1yl)ethyl]pyridin-3-carbonyl]piperidin-2-yl]methoxy]phenol was obtained. LCMS (ES) [M+1] ⁺ m/z: 440.3.

Step 8: Synthesis of (S)-2-hydroxy-6-((1-(2-(2-(pyrrolidin-1-yl)ethyl)nicotinoyl)piperidin-2yl)methoxy)benzaldehyde (4).

A mixture of 2-(hydroxymethyl)-3-[[(2S)-1-[2-[2(pyrrolidin-1-yl)ethyl]pyridin-3-carbonyl]piperidin-2-yl] was placed in a 50 ml round bottom flask. methoxy]phenol (200 mg, 0.46 mmol, 1 eq.), DCM (10 ml) and MnO ₂ (791.1 mg, 9.10 mmol, 20.00 eq.). The resulting mixture was stirred for 1 hour at room temperature. The reaction mixture was filtered and concentrated. The resulting residue was purified by preparative reverse phase HPLC (Prep-d8, 5 mM XBridge column, 19x150 mm, water; gradient elution from 15% MeCN in water to 35% MeCN in water over a 6-minute period when both solvents contained 0.1% trifluoroacetic acid) to obtain 2-hydroxy-6-[[(2S)-1-[2-[2-(pyrrolidin-1-yl)ethyl]pyridin-3-carbonyl]piperidin-2-yl] methoxy]benzaldehyde. ¹ H NMR (300 MHz, DMSO-d ₆ ) δ 11.57 (s, 1H), 10.26 (s, 1H), 9.48 (s, 1H), 8.60 (dd, J= 4, 8, 1.8 Hz, 1H), 7.65 (s, 1H), 7.53 (t, J= 8.3 Hz, 1H), 7.38 (dd, J= 7.7, 4.8 Hz, 1H), 6.73 (s, 1H), 6.61-6.52 (m, 1H), 4.61-4.41 (m, 1H), 4.41-4.25 (m, 1H), 3.60 (t, J= 7.2 Hz, 2H), 3.51-2.96 (m, 8H), 2.07-1.84 (m, 5H), 1.81-1 .36 (m, 5N). LCMS (ES) [M+1] ⁺ m/z: 438.2.

Example 5. (S)-2-Hydroxy-6-((1-(2-(hydroxymethyl)benzoyl)piperidin-2-yl)methoxy)benzaldehyde, compound 5.

Compound 5 was synthesized according to Scheme 5.

- 47 045055

Scheme 5

In Scheme 5, TBDMSCl refers to tert-butyldimethylsilyl chloride and MsCl refers to mesyl chloride. Compound 5: MS m/z 370.2 [M+H]+, 392.2 [M+Na]+.

Example 6. (R)-2-G hydroxy-6-((4-(2-(2-hydroxyethyl)nicotinoyl)thiomorpholin-3 yl)methoxy)benzaldehyde and (S)-2-hydroxy-6-((4 -(2-(2-hydroxyethyl)nicotinoyl)thiomorpholin-3yl)methoxy)benzaldehyde.

Scheme 6A

Connection 10

Enantiomers 1 and 2

Step 1. Synthesis of [4-(2-[2-[(tert-butyldimethylsilyl)oxy]ethyl]pyridin-3-carbonyl)thiomorpholin3-yl]methanol (6a).

Add 2-[2-[(tertbutyldimethylsilyl)oxy]ethyl]pyridine-3-carboxylic acid (2.00 g, 7.11 mmol, 1.00 eq.), thiomorpholin-3ylmethanol (0 .95 g, 7.13 mmol, 1.00 eq.), N,N-diisopropylethylamine (2.76 g, 21.32 mmol, 3.00 eq.) and DCM (30.00 mL). To this mixture was added in portions at 0°C 1-[bis-(dimethylamino)methylene]-III-1,2,3-triazolo[4,5b]pyridinium-3-oxide-hexafluorophosphate (3.24 g, 8.53 mmol, 1.20 eq.). The resulting reaction mixture was allowed to warm to room temperature and stirred overnight. The reaction was then quenched by adding 30 ml of water. The resulting solution was extracted with 3x30 ml dichloromethane, the organic layers were separated, combined, dried over anhydrous sodium sulfate and concentrated. The residue was applied to a silica gel column using THF/petroleum ether (PE) (30%) as eluent. The combined fractions were concentrated to give [4-(2-[2-[(tert-butyldimethylsilyl)oxy]ethyl]pyridin-3-carbonyl)thiomorpholin-3-yl]methanol. LCMS (ES) [M+H]+ m/z: 397.

- 48 045055

Step 2. Synthesis of 2-[[4-(2-[2-[(tert-butyldimethylsilyl)oxy]ethyl]pyridin-3carbonyl)thiomorpholin-3-yl]methoxy]-6-hydroxybenzaldehyde (6b).

[4-(2-[2-[(tertbutyldimethylsilyl)oxy]ethyl]pyridin-3-carbonyl)thiomorpholin-3-yl]methanol (1.50 g, 3.78 mmol, 1 .00 eq), 2,6-dihydroxybenzaldehyde (0.63 g, 4.56 mmol, 1.21 eq), PPh ₃ (1.19 g, 4.54 mmol, 1.20 eq) and DCM (30.00 ml). Diisopropyl azodicarboxylate (0.92 g, 4.54 mmol, 1.20 eq.) was added dropwise to this solution over 20 minutes with stirring at 0°C. The resulting mixture was stirred overnight at room temperature and concentrated. The residue was applied directly to a silica gel column using THF/petroleum ether (25%) as eluent. The combined fractions were concentrated to give 2-[[4-(2-[2-[(tert-butyldimethylsilyl)oxy]ethyl]pyride in-3carbonyl)thiomorpholin-3-yl]methoxy]-6-hydroxybenzaldehyde. LCMS (ES) [M+H]+ m/z: 517.

Step 3: Separation of compound 6b using chiral-HPLC.

The racemate was purified by chiral-HPLC to obtain Enantiomer 1 and Enantiomer 2 of compound 6b under the following conditions: column, Lux Cellulose-4, 4.6x100 mm, 3 μm; mobile phase, A: n-hexane, B: ethanol (35% B for 18 minutes); flow rate: 30 ml/min; detector, 254. LCMS (ES) [M+H]+ m/z: 517 (for both compounds).

Stage 4a. Removal of tert-butyl(chloro)dimethylsilyl group to obtain compound 10, enantiomer 1 HCl (~2M) in 5 ml ethyl acetate (EA) was added dropwise to enantiomer 1 of compound 6b (335.00 mg, 0.65 mmol, 1) with stirring. .00 eq.) in ethyl acetate (3.00 ml) at 0°C. The resulting solution was stirred for 2 hours at room temperature. The pH value of the solution was adjusted to 8 with a saturated NaHCO ₃ solution. The resulting solution was extracted with 3x10 ml ethyl acetate and the organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by preparative-HPLC under the following conditions: column, XBridge Prep C18 OBD column, 19 cm, 150 mm, 5 µm; mobile phase, water (0.1% HCOOH) and acetonitrile (30% to 40% phase B for 10 minutes); detector, 254. Enantiomer 1 of compound 10 was obtained with retention time = 4.06 min. LCMS (ES) [M+H]+ m/z: 403.1; [M+Na]+ m/z: 425.1.

Stage 4b. Removal of tert-butyl(chloro)dimethylsilyl group to obtain compound 10, enantiomer 2 HCl (~2M) in 5 ml ethyl acetate was added dropwise to enantiomer 2 of compound 6b (335.00 mg, 0.65 mmol, 1.00 eq) with stirring .) in ethyl acetate (3.00 ml) at 0°C. The resulting solution was stirred for 2 hours at room temperature. The pH value of the solution was adjusted to 8 with a saturated NaHCO ₃ solution. The resulting solution was extracted with 3x10 ml ethyl acetate and the organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The crude product (200 mg) was purified by preparative-HPLC under the following conditions: column, XBridge Prep C18 OBD column, 19 cm, 150 mm, 5 µm; mobile phase, water (0.1% HCOOH) and acetonitrile (30% to 40% phase B for 10 minutes); detector, 254. Enantiomer 2 of compound 10 was obtained with retention time = 5.40 min. LCMS (ES) [M+H]+ m/z: 403.2; [M+Na]+ m/z: 425.1.

Alternative synthesis of (R)-2-hydroxy-6-((4-(2-(2-hydroxyethyl)nicotinoyl)thiomorpholin-3yl)methoxy)benzaldehyde (R)-2-hydroxy-6-((4-(2- (2-Hydroxyethyl)nicotinoyl)thiomorpholin-3yl)methoxy)benzaldehyde can be prepared directly from chiral (R)-thiomorpholin-3ylmethanol, as shown in Scheme 6B.

- 49 045055

Circuit 6B

Stage 1.

To a solution of L-cysteine (100.0 g, 825.4 mmol,

1.0 eq.) in H2O (1.0 l) was added NaOH (3.3 g,

82.5 mmol, 0.1 eq.). After cooling the reaction mixture to 0°C, ethylene oxide (100.0 g, 2.26 mol, 2.75 eq.) was added dropwise. The resulting solution was stirred for 3 hours at 0-25°C in a water/ice bath. The resulting solution was extracted with 3x500 ml ethyl acetate to remove unchanged ethylene oxide. The aqueous layer was concentrated under reduced pressure. The crude product was triturated with EtOH (200 ml) for 1 hour and filtered. Hydroxyethylcysteine was obtained. LCMS (ES) [M+1]+ m/z: 166.2; retention time 0.174 min. 1H NMR: (300 MHz, D2O, ppm): δ 3.83 (dd, J= 3.0, 6.0 Hz, 1H), 3.67 (t, J= 6.0, 2H) , 3.04 (dd, J= 14.8, 4.4 Hz, 1H), 2.97 (dd, J= 14.8, 7.4 Hz, 1H), 2.68 (t, J= 6 ,0, 2H).

Stage 2.

To a mixture of hydroxyethylcysteine (130.0 g, 786.8 mmol, 1.0 eq.) and KHCO3 (165.4 g, 1.65 mol, 2.1 eq.) in dioxane (700 ml, 8.26 mol, 10.5 eq.) and H2O (700 ml), benzyl chloroformate (CbzCl; 147.6 g, 865 mmol, 1.1 eq.) was added dropwise at 0°C for 30 minutes. The resulting solution was stirred for 5 hours at 0-25°C. The solvents were evaporated and the residue was dissolved in DMF (1000 ml). Benzyl bromide (BnBr; 148 g, 0.86 mol, 1.1 eq.) was added and the resulting mixture was stirred for 16 hours at 0-25°C. The reaction was then quenched by adding 1000 ml of water. The resulting solution was extracted with 3x1000 ml EtOAc, the combined organic layer was washed with brine, dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was applied to a silica gel column with ethyl acetate/petroleum ether (1:50 to 1:5). Benzyl(2R)-2-[[(benzyloxy)carbonyl]amino]-3-[(2-hydroxyethyl)sulfanyl]propanoate was obtained. LCMS (ES) [M+1]+ m/z: 390.5; retention time 1.146 min, 1H NMR: (300 MHz, _CDCl3 , ppm): δ 7.39-7.33 (m, 10H), 5.83 (broad s, 1H), 5.26 (d , J= 4.7 Hz, 2H), 5.17 (s, 2H), 4.71-4.65 (m, 1H), 3.69-3.63 (m, 2H), 3.09- 2.98 (m, 2H), 2.71-2.61 (m, 2H).

Stage 3.

Benzyl (2R)-2-[[(benzyloxy)carbonyl]amino]-3-[(2hydroxyethyl)sulfanyl]propanoate (90.0 g, 231 mmol, 1.0 eq.), THF (1.0 l), diethyl azodicarboxylate (DEAD; 48.3 g, 277 mmol, 1.2 eq.). After cooling the reaction mixture to 0°C, PPh ₃ (78.8 g, 300 mmol, 1.3 eq.) in THF (100 ml) was added dropwise. The resulting solution was stirred for 16 hours at 0-25°C. The resulting mixture was concentrated. The residue was applied to a silica gel column with THF/petroleum ether (1:100 to 1:5). 3,4-dibenzyl-(3R)thiomorpholine-3,4-dicarboxylate was obtained. LCMS (ES) [M+1]+ m/z: 372.1; Retention time 1.312 min.

Stage 4.

3,4-Dibenzyl-(3R)-thiomorpholine-50 045055

3,4-dicarboxylate (100.0 g, 269 mmol, 1.0 eq), DCM (1.0 L). The reaction mixture was cooled to 0°C and trimethylsilyl iodide (161.6 g, 0.81 mol, 3 eq.) was added dropwise. The resulting solution was stirred for 1 hour at 0-25°C in a water/ice bath. The reaction was then quenched by adding 100 mL of MeOH. The resulting mixture was concentrated.

The pH of the solution was adjusted to 1 using HCl (2 mol/L). The resulting solution was extracted with 2x500 ml MTBE (methyl tert-butyl ether) and the aqueous layers were combined. NaHCO ₃ (2 mol/l) was used to adjust the pH to 8. The resulting solution was extracted with 3x500 ml ethyl acetate. The combined organic layer was dried over Na ₂ SO ₄ and concentrated under reduced pressure. Benzyl-(3R)-thiomorpholine-3-carboxylate was obtained. LCMS (ES) [M+1]+ m/z: 238.1; retention time 1.026 min; 1H NMR: (300 MHz, CDCl ₃ , ppm): δ 7.43-7.33 (m, 5H), 5.25 (s, 2H), 3.74 (dd, J= 8.6 , 3.4 Hz, 1H), 3.40 (ddd, J= 12.5, 4.9, 3.0 Hz, 1H), 3.04 (ddd, J= 12.5, 9.8, 2 .7 Hz, 1H), 2.90 (dd, J = 13.2, 3.4, 1.3 Hz, 1H), 2.82 (dd, J = 13.3, 8.6 Hz, 1H) , 2.70 (dddd, J= 12.9, 9.8, 3.0 Hz, 1H), 2.48 (dddd, J= 13.3, 4.9, 2.7, 1.3 Hz, 1H).

Stage 5.

To a suspension of LiAlH ₄ (13.2 g, 347 mmol, 1.5 eq.) in THF (1000 ml), benzyl (3R)thiomorpholine-3-carboxylate (55.0 g, 231.7 mmol, 1 .0 eq.) in THF (100 ml) at 0°C. After stirring the resulting solution for 3 hours at 0-25°C, the reaction was quenched by adding 100 g of Na ₂ SO ₄ -10H ₂ O. The resulting solution was diluted with 500 ml of THF and the solids were filtered off. The resulting mixture was concentrated and the residue was applied to a THF/petroleum ether (1:50 to 2:1) silica gel column. (3R)-thiomorpholin-3-ylmethanol was obtained. LCMS (ES) [M+1] ⁺ m/z: 134.1; retention time 0.464 min; 1H NMR: (300 MHz, DMSO- _d6 , ppm): δ 4.64 (broad s, 1H), 3.26-3.17 (m, 2H), 2.81-2.68 ( m, 4H), 2.43-2.26 (m, 4H).

Stage 6.

2-[2-[(tert-Butyldimethylsilyl)oxy]ethyl]pyridine-3-carboxylic acid (20.00 g, 71.06 mmol, 1.00 eq.), (3R)thiomorpholine- 3-ylmethanol (10.41 g, 78.14 mmol, 1.10 eq), DCM (300.00 mL) and N,Ndiisopropylethylamine (18.37 g, 142.13 mmol, 2.00 eq). After this, 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide-hexafluorophosphate (32.43 g, 85.28 mmol, 1. 20 equiv.) at 0°C. The resulting solution was stirred for 3 hours at room temperature. The reaction was then quenched by adding 200 ml of water. The resulting solution was extracted with 3x200 ml dichloromethane, the organic layers were combined, dried over anhydrous sodium sulfate and concentrated. The residue was applied to a THF/petroleum ether (30%) silica gel column. [(3R)-4-(2-[2-[(tert-butyldimethylsilyl)oxy]ethyl]pyridin-3-carbonyl)thiomorpholin-3-yl]methanol was obtained. LCMS (ES) [M+H]+ m/z: 397.30.

Stage 7.

2,6-Dihydroxybenzaldehyde (7.52 g, 54.45 mmol, 1.20 eq.), [(3R)-4-( 2-[2-[(tert-butyldimethylsilyl)oxy]ethyl]pyridine-3-carbonyl)thiomorpholin-3yl]methanol (18.00 g, 45.38 mmol, 1.00 eq.), PPh ₃ (14.28 g, 54.46 mmol, 1.20 eq.) and DCM (400.00 ml). Diisopropyl azodicarboxylate (11.01 g, 54.46 mmol, 1.20 eq) was then added dropwise with stirring at 0°C. The resulting solution was stirred overnight at room temperature. The resulting mixture was concentrated. The residue was applied to a THF/petroleum ether (15%) silica gel column. 2-[[(3R)-4-(2-[2-[(tert-butyldimethylsilyl)oxy]ethyl]pyridin-3carbonyl)thiomorpholin-3-yl]methoxy]-6-hydroxybenzaldehyde was obtained. LCMS (ES) [M+H]+ m/z: 517.35.

Stage 8.

2-[[(3R)-4-(2-[2-[(tertbutyldimethylsilyl)oxy]ethyl]pyridin-3-carbonyl)thiomorpholin-3-yl]methoxy]-6-hydroxybenzaldehyde ( 13.50 g, 26.13 mmol, 1.00 eq.) and ethyl acetate (20.00 ml). To the above was added HCl(gas) in ethyl acetate (52.25 mL, 104.50 mmol, 4.00 eq.) dropwise with stirring at 0°C. The resulting solution was stirred for 2 hours at room temperature. The reaction was then quenched by adding 80 ml of water. The pH value of the solution was adjusted to 7-8 with a saturated solution of Na ₂ CO ₃ . The resulting solution was extracted with 3x100 ml dichloromethane, the organic layers were combined, oven dried under reduced pressure and concentrated. The crude product was purified by preparative-HPLC under the following conditions: column, XBridge Prep C18 OBD column, 19 cm, 150 mm, 5 µm; mobile phase, water (0.1% HCOOH) and acetonitrile (30% to 50% phase B for 11 minutes); detector, 254. 2-Hydroxy-6-[[(3R)-4-[2-(2-hydroxyethyl)pyridin-3-carbonyl]thiomorpholin-3yl]methoxy]benzaldehyde was obtained.

Chiral HPLC conditions were as follows: instrument: SHIMADZU LC-20AT; mobile phase A: n-hexane (0.1% trifluoroacetic acid); mobile phase B: ethanol; conc. phase B: 50.0%; flow rate: 1,000 ml/min; column: Lux Cellulose-4, 4.6*100 mm, 3 µm. Chiral HPLC retention time = 5.41 min.

LCMS (ES, m/z): [M+H]+: 403.2; 1H NMR (300 MHz, DMSO^): δ 11.80-11.73 (m, 1H), 10.33 (broad 51 045055 s, 1H), 8.56 (dd, J = 4.9, 1.8 Hz, 1H), 7.90-7.39 (m, 2H), 7.37-7.19 (m, 1H), 6.81-6.63 (m, 1H), 6.56 (d, J=

8.4 Hz, 1H), 5.49-4.60 (m, 1H), 4.60-4.05 (m, 2H), 3.88-3.36 (m, 4H), 3.20 -2.61 (m, 6H), 2.43 (d, J= 12.6 Hz,

1H).

Based on the product of Scheme 6B, compound 10, enantiomer 2 was determined to correspond to (R)-2hydroxy-6-((4-(2-(2-hydroxyethyl)nicotinoyl)thiomorpholin-3-yl)methoxy)benzaldehyde.

Example 7. (S)-2-Hydroxy-6-((4-(2-(2-hydroxyethyl)nicotinoyl)morpholin-3yl)methoxy)benzaldehyde, compound 8.

Compound 8 was synthesized according to Scheme 7.

Scheme 7

Step 1. Synthesis of (R)-(2-(2-((tert-butyldimethylsilyl)oxy)ethyl)pyridin-3-yl)(3(hydroxymethyl)morpholino)methanone (7a).

To a solution of 2-[2-[(tert-butyldimethylsilyl)oxy]ethyl]pyridin-3-carboxylic acid (1.50 g, 5.33 mmol, 1.00 eq.) and (3R)-morpholin-3-ylmethanol hydrochloride (0.98 g, 6.39 mmol, 1.20 eq.) in DCM (20 ml) was added N,N-diisopropylethylamine (2.07 g, 15.99 mmol, 3.00 eq.) followed by 1-[bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide-hexafluorophosphate (2.43 g, 6.39 mmol, 1.20 eq. ) in portions over 5 minutes. The resulting solution was stirred for 2 hours at room temperature and diluted with 50 ml of H2O. The resulting solution was extracted with 2x30 ml dichloromethane and the organic layers were separated, combined, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to obtain a residue, which was purified by a silica gel column using ethyl acetate/petroleum ether (2/1) as eluent. (R)-(2-(2-((tertbutyldimethylsilyl)oxy)ethyl)pyridin-3-yl)(3-(hydroxymethyl)morpholino)methanone was obtained. LCMS (ES) [M+H]+ m/z: 381.2.

Step 2. Synthesis of (S)-2-((4-(2-(2-((tert-butyldimethylsilyl)oxy)ethyl)nicotinoyl)morpholin-3yl)methoxy)-6-hydroxybenzaldehyde (7b).

Solution of (R)-(2-(2-((tert-butyldimethylsilyl)oxy)ethyl)niridin-3-yl)(3-(hydroxymethyl)morpholino)methanone (600 mg, 1.57 mmol, 1.00 eq. ), 2,6-dihydroxybenzaldehyde (261 mg, 1.89 mmol, 1.20 eq.) and PPh ₃ (496 mg, 1.89 mmol, 1.20 eq.) in DCM (10 ml) were purged and maintained at inert nitrogen atmosphere. To this mixture, diisopropyl azodicarboxylate (382 mg, 1.89 mmol, 1.20 eq.) was added dropwise with stirring at 0° C. over 5 minutes. The resulting solution was stirred for 1 hour at room temperature and diluted with 20 ml of H2O. The resulting solution was extracted with 2x20 ml dichloromethane, the organic layers were combined, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to obtain the crude product, which was purified using flash preparative-HPLC under the following conditions (IntelFlash-1): column, silica gel C18; mobile phase, H2O (0.1% HCOOH)/acetonitrile (ACN) = 2/1 increasing to H2O (0.1% HCOOH)/acetonitrile = 1/4 for 18 minutes; detector, UV 254 nm. (B)-2-((4-(2-(2-((tertbutyldimethylsilyl)oxy)ethyl)nicotinoyl)morpholin-3-yl)methoxy)-6-hydroxybenzaldehyde was obtained. LCMS (ES) [M+H]+ m/z: 501.2.

Step 3. Synthesis of (S)-2-hydroxy-6-((4-(2-(2-hydroxyethyl)nicotinoyl)morpholin-3yl)methoxy)benzaldehyde (S).

Formic acid (HCOOH, 1 ml) was added to a solution of (S)-2-((4-(2-(2-((tertbutyldimethylsilyl)oxy)ethyl)nicotinoyl)morpholin-3-yl)methoxy)-6-hydroxybenzaldehyde ( 450 mg, 0.89 mmol, 1.00 eq.) in acetonitrile (5.00 ml). The resulting solution was stirred for 3 hours at 40°C, cooled to room temperature and diluted with 5 ml of acetonitrile. The mixture was concentrated to obtain the crude product, which was purified by preparative-HPLC under the following conditions (2#SHIMADZU (HPLC-01)): column, Atlantis HILIC OBD column, 19*150mm*5μm;

mobile phase, water (0.1% formic acid and acetonitrile (from 37% to 45% phase B for 10 minutes); detector, UV 254 nm. Received (S)-2-hydroxy-6-((4-( 2-(2-hydroxyethyl)nicotinoyl)morpholin3-yl)methoxy)benzaldehyde LCMS (ES) [M+H]+ m/z: 387.1.

Example

8.

(S)-2-r hydroxy-6-(( 1 -(2-(hydroxymethyl)nicotinoyl)piperidin-2 yl)methoxy)benzaldehyde, compound 11.

Compound 11 was synthesized according to Scheme 8.

Scheme 8

Step 1. Synthesis of 3-(methoxycarbonyl)-2-methylpyridine-1-oxide.

3-chloroperbenzoic acid (m-CPBA, 34, 4 g, 199.34 mmol, 2.0 eq.). The reaction mixture was stirred for 2 hours at room temperature. The reaction was quenched with saturated aqueous Na ₂ CO ₃ solution (100 ml), the organic phase was separated and dried over anhydrous sodium sulfate. The solution was filtered and the filtrate was concentrated under reduced pressure. The residue was purified on a silica gel column with dichloromethane/methanol (10/1) to give 3-(methoxycarbonyl)-2methylpyridine-1-oxide. LCMS (ES) [M+H]+ m/z: 168.

Stage 2. Synthesis of methyl 2-(acetoxymethyl)nicotinate.

A mixture of 3-(methoxycarbonyl)-2-methylpyridine-1-oxide (8.00 g) in acetic anhydride (80 ml) was heated for 5 hours at 140°C. After cooling to room temperature, excess liquid was removed under reduced pressure, the residue was suspended in water (50 ml) and extracted with 3x50 ml dichloromethane. The combined organic phase was dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The residue was purified on a silica gel column with ethyl acetate/petroleum ether (15%) to give methyl 2-(acetoxymethyl)nicotinate. LCMS (ES) [M+H]+ m/z: 210.

Stage 3. Synthesis of methyl 2-(hydroxymethyl)nicotinate.

To a solution of methyl 2-(acetoxymethyl)nicotinate (7.90 g, 37.76 mmol, 1.0 eq.) in MeOH (80 ml) was added acetyl chloride (3.60 g, 45.86 mmol, 1.2 eq. .). The reaction solution was stirred overnight at room temperature; then the solvent was removed under reduced pressure and the resulting residue was dissolved in water (20 ml). The pH was adjusted to 8 with solid NaHCO ₃ and extracted with ethyl acetate (30 ml*3). The combined organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated in vacuo. The residue was purified on a silica gel column with ethyl acetate/petroleum ether (1/1) to give methyl 2-(hydroxymethyl)pyridine-3-carboxylate. LCMS (ES) [M+1]+ m/z: 168.

- 53 045055

Step 4. Synthesis of methyl 2-(((tert-butyldimethylsilyl)oxy)methyl)nicotinate.

A 100 mL three-neck round bottom flask was filled with methyl 2-(hydroxymethyl)pyridine-3-carboxylate (2.80 g, 16.75 mmol, 1.0 eq), DCM (40 mL), and imidazole (2.27 g, 33 .34 mmol, 2.0 eq.). Thereafter, tert-butyldimethylchlorosilane (4.04 g, 26.81 mmol, 1.6 eq.) was added at 0°C. The mixture was stirred for 2 hours at room temperature. The reaction was then quenched by adding water (30 ml) and extracted with 3x50 ml of dichloromethane. The combined organic phase was dried over anhydrous sulfate and filtered, and the filtrate was concentrated in vacuo. The residue was purified on a silica gel column with ethyl acetate/petroleum ether (10%) to give methyl 2-(((t-butyldimethylsilyl)oxy)methyl)nicotinate. LCMS (ES) [M+1]+ m/z: 282.

Stage 5. Synthesis of 2-(((tert-butyldimethylsilyl)oxy)methyl)nicotinic acid.

Methyl 2-(((tertbutyldimethylsilyl)oxy)methyl)nicotinate (4.20 g, 14.92 mmol, 1.0 eq.), MeOH (30 ml) and H ₂ O ( 15 ml). LiOH-H ₂ O (1.25 g, 29.79 mmol, 2.0 eq.) was then added at 0°C. The mixture was stirred for 2 hours at room temperature, then concentrated to remove the solvent, and the pH of the residue was adjusted to pH 7 with citric acid. The solution was filtered and the solid was dried under an infrared lamp. 2-(((tert-butyldimethylsilyl)oxy)methyl)nicotinic acid was obtained. LCMS (ES) [M+1] ⁺ m/z: 268.

Step 6. Synthesis of (S)-(2-(((tert-butyldimethylsilyl)oxy)methyl)pyridin-3-yl)(2(hydroxymethyl)piperidin-1-yl)methanone.

2-(((tert-butyldimethylsilyl)oxy)methyl)nicotinic acid (615 mg, 2.30 mmol, 1.0 eq.), (2S)-piperidin-2ylmethanol (318 mg, 2.0 eq.) was placed in a 50 mL three-neck round-bottom flask. 76 mmol, 1.2 eq.), DCM (10 ml), N,N-diisopropylethylamine (594 mg, 4.60 mmol, 2.0 eq.). 1-[Bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3oxide-hexafluorophosphate (1.05 g, 2.76 mmol, 1.2 eq) was then added .) at 0°C. The mixture was stirred for 2 hours at room temperature. The reaction mixture was concentrated to remove the solvent and the residue was purified on a silica gel column with ethyl acetate/petroleum ether (80%). (S)-(2(((tert-butyldimethylsilyl)oxy)methyl)pyridin-3-yl)(2-(hydroxymethyl)piperidin-1-yl)methanone was obtained. LCMS (ES) [M+1] ⁺ m/z: 365.

Step 7. Synthesis of (S)-2-((1-(2-(((tert-butyldimethylsilyl)oxy)methyl)nicotinoyl)piperidin-2yl)methoxy)-6-hydroxybenzaldehyde.

(S)-(2-(((tert-butyldimethylsilyl)oxy)methyl)pyridin-3-yl)(2(hydroxymethyl)piperidin-1 -yl)methanone (316 mg, 0.87 mmol, 1.0 eq.), 2,6-dihydroxybenzaldehyde (143 mg, 1.04 mmol, 1.2 eq.), PPh ₃ (340 mg, 1.30 mmol, 1.5 eq.) and THF (15 ml). Diisopropyl azodicarboxylate (262 mg, 1.30 mmol, 1.5 eq.) was then added at 0°C. After addition, the reaction solution was stirred overnight at room temperature and then concentrated to remove the solvent. The resulting residue was purified on a silica gel column with ethyl acetate/petroleum ether (1/1). (S)-2-((1-(2-(((tert-butyldimethylsilyl)oxy)methyl)nicotinoyl)piperidin-2yl)methoxy)-6-hydroxybenzaldehyde was obtained. LCMS (ES) [M+1] ⁺ m/z: 485.

Step 8. Synthesis of (S)-2-hydroxy-6-((1-(2-(hydroxymethyl)nicotinoyl)piperidin-2yl)methoxy)benzaldehyde.

(S)-2-((1-(2-(((tertbutyldimethylsilyl)oxy)methyl)nicotinoyl)piperidin-2-yl)methoxy)-6-hydroxybenzaldehyde (250 mg, 0.52) was placed in a 50 ml round bottom flask. mmol, 1.0 eq.) in ethyl acetate (3 ml). HCl (gas) (2M in ethyl acetate) (5.0 ml) was added to the above solution at 0°C, and the mixture was left stirring for 1 hour at room temperature. The reaction mixture was then diluted by adding water (20 ml), the solution was adjusted to pH 8 with solid NaHCO ₃ and extracted with 3x20 ml ethyl acetate. The combined organic phase was concentrated under reduced pressure and the residue was purified by preparative-HPLC under conditions: (2#SHIMADZU(HPLC-01)): column, Kinetex EVO C18 column, 21.2x150, 5 µm, mobile phase, water. (0.1% formic acid) and CH ₃ CN (10% to 90% phase B for 15 minutes), detector, UV 254 nm. (S)-2-hydroxy-6-((1-(2-(hydroxymethyl)nicotinoyl)piperidin-2yl)methoxy)benzaldehyde was obtained. LCMS-pH (ES, m/z): [M+H]+: 371.1; [M+Na]+: 393.1.

Example 9. (S)-2-Hydroxy-6-((4-(2-(hydroxymethyl)nicotinoyl)morpholin-3-yl)methoxy)benzaldehyde, compound 12.

Compound 12 was synthesized according to Scheme 9A.

- 54 045055

Scheme 9A

Step 1. Synthesis of (R)-(2-(((tert-butyldimethylsilyl)oxy)methyl)pyridin-3-yl)(3(hydroxymethyl)morpholino)methanone.

To a solution of 2-[[(tert-butyldimethylsilyl)oxy]methyl]pyridine-3-carboxylic acid (530 mg, 1.98 mmol, 1.0 eq.), (3R)-morpholin-3-ylmethanol hydrochloride (364 mg , 2.38 mmol, 1.2 eq.) and N,Ndiisopropylethylamine (768 mg, 5.94 mmol, 3.0 eq.) in DCM (10 ml) was added 1-[bis(dimethylamino)methylene]-1H- 1,2,3-triazolo[4,5-b]pyridinium-3-oxide-hexafluorophosphate (905 mg, 2.38 mmol, 1.2 eq.) at 0°C. The reaction solution was stirred for 3 hours at room temperature. The solution was then concentrated to remove the solvent and the residue was purified on a silica gel column with ethyl acetate/petroleum ether (60%). (R)-(2-(((tertbutyldimethylsilyl)oxy)methyl)pyridin-3-yl)(3-(hydroxymethyl)morpholino)methanone was obtained. LCMS (ES) [M+1]+ m/z: 367.

Step 2. Synthesis of (S)-2-((4-(2-(((tert-butyldimethylsilyl)oxy)methyl)nicotinoyl)morpholin-3yl)methoxy)-6-hydroxybenzaldehyde.

(R)-(2-(((tert-butyldimethylsilyl)oxy)methyl)pyridin-3-yl)(3-(hydroxymethyl)morpholino)methanone was placed in a 40 ml vial, which was purged and inside which an inert atmosphere of nitrogen was maintained. (630 mg, 1.72 mmol, 1.0 eq.), 2,6-dihydroxybenzaldehyde (284 mg, 2.06 mmol, 1.2 eq.), PPh ₃ (540 mg, 2.06 mmol, 1.0 eq.), 2 eq.), THF (20 ml). Di-tert-butyl azodicarboxylate (474 mg, 2.06 mmol, 1.2 eq.) was then added at 0°C. The reaction solution was stirred overnight at room temperature. The solution was concentrated in vacuo to remove the solvent and the residue was purified on a silica gel column with ethyl acetate/petroleum ether (1/1). (S)-2-((4-(2-(((tertbutyldimethylsilyl)oxy)methyl)nicotinoyl)morpholin-3-yl)methoxy)-6-hydroxybenzaldehyde was obtained. LCMS (ES) [M+1]+ m/z: 487.

Step 3. Synthesis of (S)-2-hydroxy-6-((4-(2-(hydroxymethyl)nicotinoyl)morpholin-3yl)methoxy)benzaldehyde.

(S)-2-((4-(2-(((tertbutyldimethylsilyl)oxy)methyl)nicotinoyl)morpholin-3-yl)methoxy)-6-hydroxybenzaldehyde (380 mg, 0.78) was placed in a 25 ml round bottom flask. mmol, 1.0 eq.). To the above was added HCl(gas) (2 M) in ethyl acetate (5 ml) at 0°C. The reaction solution was stirred for 1 hour at room temperature. The reaction was then quenched by adding water (10 ml), the solution was adjusted to pH 8 with solid NaHCO ₃ and extracted with 3 x 10 ml ethyl acetate. The combined organic phase was dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative-HPLC under the following conditions (2#SHIMADZU (HPLC-01)): column, Kinetex EVO C18 column, 21.2x150, 5 µm, mobile phase, water (0.1% formic acid) and CH3CN (10% to 50% phase B in 15 minutes), detector, UV 254 nm. (S)-2-hydroxy-6((4-(2-(hydroxymethyl)nicotinoyl)morpholin-3-yl)methoxy)benzaldehyde was obtained. LCMS: (ES, m/z): [M+H]+: 373.1; [M+Na]+: 395.1.

Alternative synthesis: circuit 9B.

Alternatively, compound 12 can be synthesized as shown in Scheme 9B using similar procedures described in Scheme 9A.

- 55 045055

Compound 9c can be converted to 9d using methods known in the art (eg, sodium tetrahydroborate; acetic acid in tetrahydrofuran at 15°C; for 4 hours). Using a silyl protecting group such as tert-butyldiphenylsilyl, intermediate 8e2 can then be converted to 12 under conditions similar to those described in Scheme 9A.

Example 10.

yl)methoxy)benzaldehyde and yl)methoxy)benzaldehyde

Scheme 10A (S)-2-Hydroxy-6-((4-(2-(hydroxymethyl)nicotunoyl)thiomorpholine-3(R)-2-hydroxy-6-((4-(2-(hydroxymethyl)nicotinoyl)thiomorpholine -3-

Enantiomers 1 and 2 of Compound 10b

Step 1. Synthesis of (2-(((tert-butyldimethylsilyl)oxy)methyl)pyridin-3-yl)(3(hydroxymethyl)thiomorpholino)methanone.

To a solution of 2-[[(tert-butyldimethylsilyl)oxy]methyl]pyridine-3-carboxylic acid (1.20 g, 4.50 mmol, 1.00 eq.) and thiomorpholin-3-ylmethanol hydrochloride (912 mg, 5 .40 mmol, 1.20 eq.) N,N-diisopropylethylamine (909 mg, 9.00 mmol, 2.00 eq.) was added to DMF under a nitrogen atmosphere at 0°C. After this, 1-[bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5b]pyridinium-3-oxide-hexafluorophosphate (2.05 g, 5.40 mmol, 1. 20 equiv.) at 0°C. The mixture was allowed to slowly warm to room temperature and stirred for 16 hours. The reaction mixture was diluted with water (100 ml) and extracted with 3x100 ml ethyl acetate. The combined organic layer was washed with 3 times 50 ml of brine, dried over anhydrous sodium sulfate and concentrated. The residue was applied to a silica gel column and eluted with ethyl acetate/petroleum ether (0% to 100% ethyl acetate). Removal of solvents resulted in [4-(2-[[(tertbutyldimethylsilyl)oxy]methyl]pyridin-3-carbonyl)thiomorpholin-3-yl]methanol. LCMS (ES) [M+1]+ m/z: 383.

- 56 045055

Stage 2. Synthesis of 2-((4-(2-(((tert-butyldimethylsilyl)oxy)methyl)nicotinoyl)thiomorpholin-3yl)methoxy)-6-hydroxybenzaldehyde.

To a mixture of [4-(2-[[(tert-butyldimethylsilyl)oxy]methyl]pyridine-3-carbonyl)thiomorpholin-3yl]methanol (1.20 g, 3.14 mmol, 1.00 eq.), 2, 6-dihydroxybenzaldehyde (519 mg, 3.76 mmol, 1.20 eq.) and PPh ₃ (0.99 g, 3.76 mmol, 1.20 eq.) in THF (50.0 ml) under nitrogen at At 0°C, a solution of di-tert-butyl azodicarboxylate (0.87 g, 3.76 mmol, 1.20 eq) in THF (1 ml) was added dropwise over 15 minutes. The resulting mixture was stirred for 16 hours at room temperature. The reaction mixture was concentrated to remove solvents and the residue was applied to a silica gel column, eluting with ethyl acetate/petroleum ether (1:0). After removal of the solvents, this gave 2-[[4-(2-[[(tert-butyldimethylsilyl)oxy]methyl]pyridin-3-carbonyl)thiomorpholin-3-yl]methoxy]-6hydroxybenzaldehyde. LCMS (ES) [M+1]+ m/z: 503.

Step 3: Separation of compound 10b using chiral HPLC.

Racemic 2-[[4-(2-[[(tert-butyldimethylsilyl)oxy]methyl]pyridin-3-carbonyl)thiomorpholin3-yl]methoxy]-6-hydroxybenzaldehyde was purified by chiral-preparative-HPLC under the following conditions: Agela HP-Flash (model: HP-1000); mobile phase: A: n-hexane/DCM=5/1; B: ethanol; flow rate: 30 ml/min; column: CHIRALPAK IG-3, 4.6*50 mm, 3 µm; and gradient: 20% B for 15 minutes; 220 nm.

Each of enantiomers 1 and 2 of compound 10b was obtained (Rt=10 min and 12 min, respectively). LCMS (ES) [M+1]+ m/z: 503.

Stage 4a. Removal of tert-butyl(chloro)dimethylsilyl group to give compound 13, enantiomer 1.

To a solution of enantiomer 1 of 10b (119 mg, 0.24 mmol, 1.00 eq.) in THF (10.0 mL) was added triethylamine trihydrofluoride (TEA-3HF) (458 mg, 2.84 mmol, 12.0 eq. ). The reaction mixture was stirred for 16 hours at room temperature. The solvents were removed and the residue was applied to a C18 silica gel column with phase A: water/0.05% trifluoroacetic acid, mobile phase B: acetonitrile; flow rate: 1.5 ml/min; gradient: from 5% to 100% B over 1.2 min, hold 0.6 min. Compound 13 was obtained, enantiomer 1. Retention time = 3.614 min; LCMS (ES, m/z): [MH] ⁺ 389.1.

Stage 4b. Removal of tert-butyl(chloro)dimethylsilyl group to give compound 13, enantiomer 2.

To a solution of enantiomer 2 of 10b (120 mg, 0.24 mmol, 1.00 eq) in THF (10.0 mL) was added triethylamine trihydrofluoride (461 mg, 2.87 mmol, 12.0 eq). The mixture was stirred for 16 hours at room temperature. The solvents were removed and the residue was applied to a C18 silica gel column with phase A: water/0.05% trifluoroacetic acid, mobile phase B: acetonitrile; flow rate: 1.5 ml/min; gradient: from 5% to 100% B over 1.2 minutes, hold 0.6 minutes. Compound 13, enantiomer 2 was obtained. Retention time = 4.387 min; LCMS (ES, m/z): [M+H]+ 389.1; [M+Na]+411.1.

Alternative synthesis of (K)-2-hydroxy-6-((4-(2-(2-hydroxyethyl)nicotinoyl)thiomorpholin-3yl)methoxy)benzaldehyde.

Alternatively, (K)-2-hydroxy-6-((4-(2-(2-hydroxyethyl)nicotinoyl)thiomorpholin-3yl)methoxy)benzaldehyde can be prepared directly from chiral (K)-thiomorpholin-3ylmethanol as shown on the diagram 10V.

Circuit 10V

10s 1Ое

O.T.B.S.

- 57 045055

Stage 1.

A solution of (3-bromopyridin-2-yl)methanol (50 g, 0.267 mol, 1.0 eq.) in DCM (1.0 L) and N-imidazole (36.4 g, 0.534 mol, 2.0 eq.). After cooling the mixture to 0° C., tert-butyl(chloro)dimethylsilane (48.1 g, 0.320 mol, 1.2 eq.) was added in three portions. The reaction solution was warmed to room temperature and stirred for 4 hours. The reaction mixture was diluted with H ₂ O (1.0 L) and extracted with 2x500 ml DCM. The combined organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by a silica gel column with ethyl acetate/petroleum ether (1:10) to give the title compound. LCMS (ES) [M+1]+ m/z 302.

Stage 2.

A solution of 3-bromo-2-(((tertbutyldimethylsilyl)oxy)methyl)pyridine (70.0 g, 0.233 mol, 1.0 eq. ) in THF (700 ml). n-BuLi (2.5 M in hexane) (102.5 mL, 0.256 mol, 1.1 eq) was then added dropwise with stirring at -78°C. After addition, the mixture was stirred for 0.5 hour, ethyl carbonochloridate (37.8 g, 0.350 mol, 1.5 eq.) was added at the same temperature and stirred for 1 hour. The reaction was then quenched by adding 500 ml of aqueous NH ₄ Cl and extracted with 2x600 ml of ethyl acetate. The combined organic phase was dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The residue was purified on a silica gel column with ethyl acetate/petroleum ether (1:4) to give the title compound. LCMS (ES) [M+1] ⁺ m/z 296.

Stage 3.

Ethyl 2-(((tert-butyldimethylsilyl)oxy)methyl)nicotinate (38.6 g, 0.131 mol, 1.0 eq.), MeOH (400 ml), and H ₂ O (200 ml) were placed in a 1 L three-neck round bottom flask. ). LiOH-H ₂ O (11.0 g, 0.262 mol, 2.0 eq.) was then added at 0°C. The mixture was stirred for 2 hours at room temperature. The mixture was concentrated to remove the solvent, and the pH of the residue was adjusted to 7 with citric acid. The solution was filtered and the solid was dried under an infrared lamp. 2-(((tert-butyldimethylsilyl)oxy)methyl)nicotinic acid was obtained. LCMS (ES) [M+1] ⁺ m/z: 268.

Stage 4

Compound 6h was prepared as described in Scheme 6B. To a solution of 2-[[(tert-butyldimethylsilyl)oxy]methyl]pyridine-3-carboxylic acid (10.0 g, 37.3 mmol, 1.0 eq.), diisopropylethylamine (12.1 g, 93.5 mmol, 2 .5 eq.) and 1-[bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5b]pyridinium-3-oxide-hexafluorophosphate (17.06 g 44.877 mmol, 1.20 eq. .) (3R)-thiomorpholin-3-ylmethanol (4.98 g, 37.397 mmol, 1.00 eq.) was added portionwise to DMF (100 mL) at 0°C. The resulting solution was stirred for 4 hours at 0-25°C. The reaction was then quenched by adding 200 ml of water. The resulting solution was extracted with 3x200 ml ethyl acetate, dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was applied to a silica gel column with ethyl acetate/petroleum ether (1:100 to 1:10). [(3R)-4-(2-[[(tert-butyldimethylsilyl)oxy]methyl]pyridin-3-carbonyl)thiomorpholin-3-yl]methanol was obtained. LCMS (ES) [M+1] ⁺ m/z: 383.2; retention time 1.138 min. 1H NMR: (300 MHz, CDCR ppm): δ 8.60 (dd, J= 4.8,1.5 Hz, 1H), 7.67-7.53 (m, 1H), 7.297, 25 (m, 1H), 5.37-4.90 (m, 2H), 4.86-4.74 (m, 1H), 4.38-4.22 (m, 1H), 3.90- 3.61 (m, 1H), 3.58-3.42 (m, 2H), 3.25-3.12 (m, 1H), 2.94-2.39 (m, 4H), 0. 96-0.88 (m, 9H), 0.21-0.01 (m, 6H).

Stage 5.

[(3R)-4-(2-[[(tert-butyldimethylsilyl)oxy]methyl]pyridin-3-carbonyl)thiomorpholin-3yl]methanol solution (11.0 g, 28.7 mmol, 1.0 eq.) , 2,6-dihydroxybenzaldehyde (4.7 g, 34.5 mmol, 1.2 eq.) and PPh ₃ (9.8 g, 37.3 mmol, 1.3 eq.) in DCM (1.1 L ) was cooled to 0°C in an Ar atmosphere. A solution of di-tert-butyl azodicarboxylate (7.28 g, 230.2 mmol, 1.1 eq) in DCM (100 ml) was added dropwise. The resulting solution was stirred for 16 hours at 0-25°C. The reaction was concentrated under vacuum. The residue was applied to a silica gel column with ethyl acetate/petroleum ether (1:100 to 1:5). 2-[[(3R)-4-(2-[[(tert-butyldimethylsilyl)oxy]methyl]pyridine-3-carbonyl)thiomorpholin-3-yl]methoxy]6-hydroxybenzaldehyde was obtained. LCMS (ES) [M+1] ⁺ m/z: 503.2; retention time 1.223 min. 1H NMR: (300 MHz, DMSO- _d6 , ppm): δ 11.74 (broad s, 1H), 10.24 (broad s, 1H), 8.59 (dd, J= 4.9 , 1.7 Hz, 1H), 7.887.41 (m, 3H), 6.76-6.54 (m, 2H), 5.44-5.32 (m, 1H), 4.90-4, 44 (m, 4H), 3.37-3.18 (m, 2H), 3.22-2.69 (m, 4H), 0.89-0.72 (m, 9H), 0.13- 0.11 (m, 6H).

Stage 6.

2-[[(3R)-4-(2-[[(tertbutyldimethylsilyl)oxy]methyl]pyridin-3-carbonyl)thiomorpholin-3-yl]methoxy]-6-hydroxybenzaldehyde (13) was placed in a 500 ml three-neck round bottom flask .6 g, 27.0 mmol, 1.0 eq.) and THF (150 ml). After cooling the reaction mixture to 0°C, a solution of triethylamine trihydrofluoride (13.0 g, 80.9 mmol, 3.0 eq.) was added dropwise. The resulting solution was stirred for 5 hours at 0-25°C. The pH of the solution was adjusted to 8 using NaHCO ₃ (2 mol/l). The resulting solution was extracted with ethyl acetate (200 mlx3), the organic layers were combined and

- 58 045055 concentrated. The crude product was purified by flash preparative-HPLC under the following conditions (IntelFlash-1): column, silica gel C18; mobile phase, MeCN=10/90, increasing to MeCN=90/10; detector, 220. 2-hydroxy-6-[[(3R)-4-[2-(hydroxymethyl)pyridine-3carbonyl]thiomorpholin-3-yl]methoxy]benzaldehyde was obtained. LCMS (ES, m/z): [M+H]+: 389.1; retention time 1.060 min.

Retention time for analytical chromatography with supercritical mobile phase (Supercritical fluid chromatography, SFC): 3.641 min. The conditions for supercritical mobile phase chromatography were as follows: instrument name: Shimadzu LC30AD SF; column: OD-3, 100*3.0 mm, 3 µm; Column ID: OD3SCK-TG002; oven temperature: 35°C; total flow: 2.5000 ml/min; initial conc. pump B: 10.0%; Back Pressure Regulator (BPR) pressure: 15.00 MPa.

1H NMR (300 MHz, DMSO- _d6 , ppm): δ 11.77 (broad s, 1H), 10.30 (broad s, 1H), 8.54 (dd, J= 4.8, 1.5 Hz, 1H), 7.76-7.36 (m, 3H), 6.75-6.52 (m, 2H), 5.45-4.07 (m, 6H), 3.46 -2.72 (m, 5H), 2.51-2.39 (m, 1H).

Based on the product of Scheme 10B, compound 13, enantiomer 1 was determined to correspond to (R)-2hydroxy-6-((4-(2-(2-hydroxyethyl)nicotinoyl)thiomorpholin-3-yl)methoxy)benzaldehyde.

Example 11. (S)-2-Hydroxy-6-(( 1 -(2-(2-methoxyethyl)benzyl)piperidin-2-yl)methoxy)benzaldehyde, compound 14.

Compound 14 was synthesized according to Scheme 11.

Scheme 11

14Step 1. Synthesis of 1-bromo-2-(2-methoxyethyl)benzene.

2-(2-bromophenyl)ethanol (10.0 g, 49.7 mmol, 1.00 eq.) and DMF (100 mL) were placed in a 500 mL three-neck round bottom flask, purged and maintained under an inert nitrogen atmosphere. , cooled to 0°C with ice water, and then NaH (2.39 g, 99.5 mmol, 2.00 eq.) was added in several portions. The resulting solution was stirred for 40 minutes at 0°C, and then MeI (10.59 g, 74.610 mmol, 1.50 eq) was added dropwise with stirring at 0°C for 15 minutes. The resulting solution was allowed to warm to room temperature with stirring for an additional 5 hours. The reaction was then quenched by adding water/ice. The resulting solution was extracted with 3x100 ml ethyl acetate, the organic layers were combined, dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column chromatography using ethyl acetate/hexane (1:3) as eluent. This resulted in the production of 1-bromo-2-(2methoxyethyl)benzene. LCMS (ES) [M+1]+ m/z: 215.

Stage 2. Synthesis of ethyl 2-(2-methoxyethyl) benzoate.

1-Bromo-2-(2-methoxyethyl)benzene (10.0 g, 46.5 mmol, 1.00 eq) and THF were placed in a 500 mL three-neck round bottom flask, purged and maintained under an inert atmosphere of nitrogen. . (100 ml). The mixture was cooled to -78°C and n-butyllithium (39 mL, 97.7 mmol, 2.10 eq.) was added dropwise to the solution. The resulting solution was stirred for 40 minutes at -78°C, then ethyl chloroformate (7.57 g, 69.757 mmol, 1.50 eq.) was added dropwise. The resulting solution was brought to room temperature with stirring for an additional 5 minutes at -78°C, and then stirred for an additional 16 hours at room temperature. The reaction was then quenched by adding water/ice. The resulting solution was extracted with 3x100 ml ethyl acetate, the organic layers were combined and dried over anhydrous sodium sulfate. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1:5) as eluent. Ethyl 2(2-methoxyethyl)benzoate was obtained. LCMS (ES) [M+1]+ m/z: 209.

Stage 3. Synthesis of 2-(2-methoxyethyl)benzoic acid.

Place ethyl 2-(2-methoxyethyl)benzoate (1.20 g, 5.76 mmol, 1.00 eq), LiOH (0.55 g, 23.0 mmol, 4.00 eq) into a 100 mL round bottom flask .), THF (15.0 ml.) and H ₂ O (3.00 ml). The resulting solution was stirred for 4 hours at 50°C in an oil bath. The reaction mixture was cooled in a water/ice bath. The pH of the solution was adjusted to 5 using HCl (2M). The resulting solution was extracted with 3x50 ml ethyl acetate, the organic layers were combined, dried over anhydrous sodium sulfate and concentrated in vacuo. 2-(2-methoxyethyl)benzoic acid was obtained. LCMS (ES) [M+1]+ m/z: 181.

Step 4. Synthesis of [(2S)-1-[2-(2-methoxyethyl)benzoyl]piperidin-2-yl]methanol.

2-(2-methoxyethyl)benzoic acid (550 mg, 3.05 mmol, 1.00 eq), (2S)-piperidin-2-ylmethanol (421 mg, 3.66 mmol, 1.20 equiv.), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide-hexafluorophosphate (2.32 g, 6.10 mmol , 2.00 eq.), N,N-diisopropylethylamine (788 mg, 6.10 mmol, 2.00 eq.) and DCM (40.00 ml). The resulting solution was stirred for 4 hours at room temperature. The resulting solution was extracted with 3x30 ml dichloromethane, the organic layers were combined, dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified on a silica gel column using ethyl acetate/petroleum ether (2:5) as eluent. [(2S)-1-[2-(2-methoxyethyl)benzoyl]piperidin2-yl]methanol was obtained. LCMS (ES) [M+1]+ m/z: 278.

Step 5. Synthesis of (S)-2-hydroxy-6-((1-(2-(2-methoxyethyl)benzoyl)piperidin-2yl)methoxy)benzaldehyde.

[(2S)-1-[2-(2-methoxyethyl)benzoyl]piperidin-2-yl]methanol (470 mg, 1.70 mmol) was placed in a 100 mL round bottom flask, which was purged and maintained under an inert nitrogen atmosphere. , 1.00 eq.), 2,6-dihydroxybenzaldehyde (468 mg, 3.39 mmol, 2.00 eq.), PPh ₃ (888 mg, 3.39 mmol, 2.00 eq.) and THF (30 ,0 ml). The resulting solution was stirred for 15 minutes at 0°C, and then diisopropyl azodicarboxylate (685 mg, 3.39 mmol, 2.00 eq.) was added dropwise. The resulting solution was stirred for 15 minutes at 0°C. The resulting solution was warmed to room temperature with stirring for an additional 16 hours. The resulting solution was extracted with 3x30 ml ethyl acetate, the organic layers were combined and dried over anhydrous sodium sulfate. The crude product was purified using preparative HPLC [column: Atlantis HILIC OBD column, 19*150mm*5μm; mobile phase: water (0.1% formic acid) and acetonitrile (10% to 50% phase B in 10 minutes, up to 90% in 10 minutes)]. (S)-2-hydroxy-6-((1-(2-(2methoxyethyl)benzoyl)piperidin-2-yl)methoxy)benzaldehyde was obtained. LCMS (ES) [M+1]+ m/z: 398. 1H NMR (300 MHz, DMSO-d ₆ ) 11.64 (broad s, 1H), 10.29 (broad s, 1H), 7.50 -6.98 (m, 5H), 6.81-6.65 (m, 1H), 6.53 (d, J= 8.4 Hz, 1H), 5.25-5.11 (m, 1H ), 4.48 (d, J= 11.3 Hz, 1H), 4.29 (dd, J= 10.2, 6.1 Hz, 1H), 3.65-3.31 (m, 2H) , 3.29-2.99 (m, 2H), 3.06 (s, 3H), 2.88-2.63 (m, 2H), 1.92-1.34 (m, 6H).

Example 12. (S)-3-(2-(2-((2-Formyl-3-hydroxyphenoxy)methyl)piperidine-1carbonyl)phenyl)propanenitrile, compound 15.

Compound 15 was synthesized according to Scheme 12.

Scheme 12

12b 1?s

12d 12e 12f

Step 1. Synthesis of methyl 2-[(1E)-2-cyanoeth-1-en-1-yl]benzoate.

Place methyl 2-bromobenzoate (5.00 g, 23.251 mmol, 1.00 eq.), acrylonitrile (12.34 g, 232.508 mmol, 10.00 eq.), N,N-diisopropylethylamine into a 100 mL round bottom flask. (6.01 g, 46.502 mmol, 2.00 eq.) and bis-(tributylphosphine) palladium (1.19 g, 2.325 mmol, 0.10 eq.). The resulting solution was stirred for 16 hours at 80°C. The resulting mixture was concentrated. The remainder was applied to

- 60 045055 silica gel column using ethyl acetate/petroleum ether (1:5) as eluent. The collected fractions were combined and concentrated. Methyl 2-[(1E)-2-cyanoeth-1-en-1-yl]benzoate was obtained. Gas chromatography-mass spectrometry (GCMS) M ⁺ : 187.

Stage 2. Synthesis of methyl 2-(2-cyanoethyl) benzoate.

A 100 mL round bottom flask was filled with methyl 2-[(1E)-2-cyanoeth-1-en-1-yl]benzoate (2.40 g, 12.8 mmol, 1.00 eq.), methanol (50% ). ml) and Pd/C (0.24 g). The flask was evacuated and purged with nitrogen three times, followed by hydrogen. The mixture was stirred for 16 hours at room temperature under a hydrogen atmosphere.

Solids were filtered off. The resulting mixture was concentrated. Methyl 2-(2cyanoethyl)benzoate was obtained. LCMS (ES) [M+1] ⁺ m/z 190.1.

Stage 3. Synthesis of 2-(2-cyanoethyl)benzoic acid.

A 100 mL round bottom flask was filled with methyl 2-(2-cyanoethyl) benzoate (2.20 g, 11.627 mmol, 1.00 eq), methanol (20 mL), water (20 mL), and sodium hydroxide (0. 93 g, 23.252 mmol, 2.00 eq.). The resulting solution was stirred for 4 hours at 25°C. The reaction was then quenched by adding 100 ml of water. The pH of the solution was adjusted to 5 using HCl (1 mol/L). Solids were collected by filtration. 2-(2-cyanoethyl)benzoic acid was obtained. LCMS (ES) [M-1]' m/z 174.1.

Step 4. Synthesis of 3-[2-[(2S)-2-[[(tert-butyldimethylsilyl)oxy]methyl]piperidine-1carbonyl]phenyl]propanenitrile.

Place 2-(2-cyanoethyl)benzoic acid (1.80 g, 10.275 mmol, 1.00 eq), DCM (30.00 mL), (2S)-2-[[(tert) into a 100 mL round bottom flask. -butyldimethylsilyl)oxy]methyl]piperidine (2.36 g, 10.275 mmol, 1.00 eq.), 1-[bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b ]pyridinium-3oxide-hexafluorophosphate (5.86 g, 15.412 mmol, 1.50 eq.) and N,N-diisopropylethylamine (3.98 g, 30.824 mmol, 3.00 eq.). The resulting solution was stirred for 16 hours at 25°C. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1:2) as eluent. The collected fractions were combined and concentrated. 3-[2-[(2S)-2-[[(tert-butyldimethylsilyl)oxy]methyl]piperidine-1carbonyl]phenyl]propanenitrile was obtained. LCMS (ES) [M+1] ⁺ m/z 387.2.

Step 5. Synthesis of 3-[2-[(2S)-2-(hydroxymethyl)piperidine-1-carbonyl]phenyl]propanenitrile.

Place 3-[2-[(2S)-2-[[(tertbutyldimethylsilyl)oxy]methyl]piperidine-1-carbonyl]phenyl]propanenitrile (2.00 g, 5.173 mmol, 1.00 equiv) in a 100 mL round bottom flask .), tetrahydrofuran (20 ml) and tetrabutylammonium fluoride (0.27 g, 1.035 mmol, 0.20 eq.). The resulting solution was stirred for 2 hours at 25°C. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1:2) as eluent. The collected fractions were combined and concentrated. 3-[2-[(2S)-2(hydroxymethyl)piperidine-1-carbonyl]phenyl]propanenitrile was obtained. LCMS (ES) [M+1] ⁺ m/z 273.2.

Step 6. Synthesis of (S)-3-(2-(2-((2-formyl-3-hydroxyphenoxy)methyl)piperidine-1carbonyl)phenyl)propanenitrile.

3-[2-[(2S)-2-(hydroxymethyl)piperidine-1carbonyl]phenyl]propanenitrile (100.00 mg, 0.367 mmol, 1.00 eq.), tetrahydrofuran (8.00 ml), 2,6dihydroxybenzaldehyde (50.72 mg, 0.367 mmol, 1.00 eq.), PPh ₃ (115.57 mg, 0.441 mmol, 1.20 eq.) and diisopropyl azodicarboxylate (89.10 mg, 0.441 mmol, 1.20 eq.). The resulting solution was stirred for 16 hours at 25°C. The resulting mixture was concentrated under vacuum. The crude reaction mixture was filtered and subjected to reverse phase preparative HPLC (Prep-C18, 20-45M, 120 g, Tianjin Bonna-Agela Technologies; gradient elution from 30% MeCN in water to 40% MeCN in water over a 10-minute period, where both solvents contained 0.1% formic acid) to obtain (S)-3-(2-(2-((2-formyl-3-hydroxyphenoxy)methyl)piperidine-1 carbonyl)phenyl)propanenitrile. LCMS (ES) [M+1] ⁺ m/z 393.2. 1H NMR (300 MHz, DMSO- _d6 ) δ 11.72 (broad s, 1H), 10.21 (m, 1H), 7.67-7.22 (m, 4H), 7.06-6, 33 (m, 2H), 5.31-5.15 (m, 1H)), 4.70-4.12 (m, 2H), 3.39-3.12 (m, 2H), 2.86 -2.67 (m, 4H), 1.93-1.25 (m, 6H).

Example 13. (S)-2-F hydroxy-6-(( 1-(3 -(2-hydroxyethyl)picolinoyl)piperidin-2-yl)methoxy)benzaldehyde, compound 16.

Compound 16 was synthesized according to Scheme 13.

- 61 045055

Scheme 13

Step 1. Synthesis of methyl 3-[(E)-2-ethoxyethenyl]pyridine-2-carboxylate.

2-[(Z)-2-ethoxyethenyl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (4, 13 g, 20.85 mmol, 1.50 eq.), methyl 3-bromopyridine-2-carboxylate (3.00 g, 13.89 mmol, 1.00 eq.), dioxane (30.00 ml), H ₂ O (6.00 ml), Na ₂ CO ₃ (4.42 g, 41.66 mmol, 3.00 eq.) and Pd(PPh ₃ ) ₄ (1.60 g, 1.39 mmol, 0 ,10 eq.). The resulting solution was stirred overnight at 80°C. The reaction mixture was cooled to room temperature. Solids were filtered off and the filtrate was concentrated. The residue was purified by silica gel column chromatography using THF/petroleum ether (30%) as eluent. Methyl 3-[(E)-2-ethoxyethenyl]pyridine-2-carboxylate was obtained. LCMS (ES) [M+1]+ m/z 208.

Step 2. Synthesis of methyl 3-(2-ethoxyethyl)pyridine-2-carboxylate.

A pre-purged 100 mL round bottom flask was charged with methyl 3-[(E)2-ethoxyethenyl]pyridine-2-carboxylate (2.60 g, 12.55 mmol, 1.00 eq), Pd/C (500.00 mg, 4.69 mmol, 0.37 eq.) and MeOH (30.00 ml). The flask was evacuated and purged with nitrogen three times, followed by hydrogen. The resulting solution was stirred overnight at room temperature. Solids were filtered off and the filtrate was concentrated. Methyl 3-(2-ethoxyethyl)pyridine 2-carboxylate was obtained. LCMS (ES) [M+1]+ m/z 210.

Stage 3. Synthesis of 3-(2-ethoxyethyl)pyridine-2-carboxylic acid.

A 100 mL round bottom flask was filled with methyl 3-(2-ethoxyethyl)pyridine-2-carboxylate (2.50 g, 11.95 mmol, 1.00 eq.), THF (25.00 mL), H2O (5. 00 ml), and LiOH-^O (1.00 g, 23.83 mmol, 1.99 eq.). The resulting solution was stirred for 2 hours at room temperature. The resulting mixture was concentrated and adjusted to pH 3-4 with HCl (1 mol/l). The resulting solution was extracted with 3x20 ml ethyl acetate, the organic layers were combined, dried over anhydrous sodium sulfate and concentrated. 3-(2-ethoxyethyl)pyridine-2-carboxylic acid was obtained. LCMS (ES) [M+1]+ m/z 196.

Step 4. Synthesis of 2-[(2S)-2-[[(tert-butyldimethylsilyl)oxy]methyl]piperidine-1-carbonyl]-3-(2ethoxyethyl)pyridine.

(28)-2-[[(tert-butyldimethylsilyl)oxy]methyl]piperidine (2.12 g, 9.22 mmol, 1.00 eq.), 3-(2-ethoxyethyl)pyridine was placed into a 100 mL three-neck round bottom flask. -2carboxylic acid (1.80 g, 9.22 mmol, 1.00 eq.), DCM (20.00 ml), Et ₃ N (1.87 g, 18.44 mmol, 2.00 eq.), 1-ethyl-3-3-dimethylaminopropylcarbodiimide (2.12 g, 11.06 mmol, 1.20 eq.) and hydroxybenzotriazole (1.50 g, 11.06 mmol, 1.20 eq.). The resulting solution was stirred overnight at room temperature. The reaction was then quenched by adding 10 ml of water. The resulting solution was extracted with 3x20 ml dichloromethane, the organic layers were combined, dried over anhydrous sodium sulfate and concentrated. 2-[(2S)-2-[[(tert-butyldimethylsilyl)oxy]methyl]piperidine-1-carbonyl]-3-(2ethoxyethyl)pyridine was obtained. LCMS (ES) [M+1]+ m/z 407.

Step 5. Synthesis of [(28)-1-[3-(2-ethoxyethyl)pyridin-2-carbonyl]piperidin-2-yl]methanol.

2-[(28)-2-[[(tert-butyldimethylsilyl)]methyl]piperidine-1-carbonyl]-3-(2-ethoxyethyl)pyridine was placed in a 100 ml round bottom flask. (3.00 g, 7.38 mmol, 1.00 eq.), THF (20.00 mL), and tetrabutylammonium fluoride/THF (14.75 mL, 14.75 mmol, 2.00 eq.). The resulting solution was stirred overnight at room temperature. The resulting mixture was concentrated. The residue was purified on a silica gel column using THF/petroleum ether (45%) as eluent. [(28)-1-[3-(2-ethoxyethyl)pyridin-2-carbonyl]piperidin-2-yl]methanol was obtained. LCMS (ES) [M+1]+ m/z 293.

Step 6. Synthesis of 2-[[(28)-1-[3-(2-ethoxyethyl)pyridin-2-carbonyl]piperidin-2-yl]methoxy]-6hydroxybenzaldehyde.

[(28)-1-[3-(2-ethoxyethyl)pyridin-2carbonyl]piperidin-2-yl]methanol (1.70 g, 5.81 mmol, 1.00 eq.) was placed into a 100 mL three-neck round bottom flask. ), 2,6-dihydroxybenzaldehyde (0.96 g,

- 62 045055

6.95 mmol, 1.20 eq.), DCM (40.00 ml) and PPh ₃ (1.83 g, 6.98 mmol, 1.20 eq.). Diisopropyl azodicarboxylate (1.41 g, 6.98 mmol, 1.20 eq) was then added dropwise with stirring at 0°C. The resulting solution was stirred for 4 hours at room temperature. The resulting mixture was concentrated. The residue was purified on a silica gel column using THF/petroleum ether (30%) as eluent. 2-[[(2S)-1-[3-(2-ethoxyethyl)pyridin-2-carbonyl]piperidin-2-yl]methoxy]-6hydroxybenzaldehyde was obtained. LCMS (ES) [M+1]+ m/z 413.

Step 7. Synthesis of (S)-2-hydroxy-6-((1-(3-(2-hydroxyethyl)picolinoyl)piperidin-2yl)methoxy)benzaldehyde.

2-[[(2S)-1-[3-(2-ethoxyethyl)pyridine2-carbonyl]piperidin-2-yl]methoxy]-6-hydroxybenzaldehyde (500.00 mg, 1. 21 mmol, 1.00 eq.) and DCM (20.00 ml). BBg ₃ /DCM (12.12 mL, 12.12 mmol, 10.00 eq) was then added dropwise with stirring at -78°C. The resulting solution was stirred for 1 hour at 0°C. The reaction was then quenched by adding 20 mL of water/ice. The resulting solution was extracted with 3x20 ml dichloromethane, the organic layers were combined, dried over anhydrous sodium sulfate and concentrated. The crude product was purified by preparative-HPLC [column, XBridge Prep C18 OBD column, 19 cm, 150 mm, 5 μm; mobile phase, water (0.1% HCOOH) and acetonitrile (30% to 60% phase B for 11 minutes); detector, 254. (S)-2-hydroxy-6-((1-(3-(2-hydroxyethyl)picolinoyl)piperidin-2yl)methoxy)benzaldehyde was obtained. LCMS: (ES, m/z): [M+1]+ 385.0. ^1H NMR (300 MHz, DMSOL) δ 11.81 (s, 1H), 10.29 (d, J= 6.6 Hz, 1H), 8.40-8.35 (m, 1H), 7, 77 (dd, J= 7.8, 1.6 Hz, 1H), 7.57-7.44 (m, 1H), 7.36 (dd, J= 7.9, 4.7 Hz, 1H) , 6.73 (d, J= 8.3 Hz, 1H), 6.63-6.46 (m, 1H), 5.20 (s, 1H), 4.68-4.40 (m, 2H ), 4.33-4.20 (m, 1H), 3.673.56 (m, 1H), 3.60-3.52 (m, 1H), 3.22-3.00 (m, 2H), 2.79-2.59 (m, 2H), 1.92-1.53 (m, 6H).

Example 14. (S)-2-Hydroxy-6-((1-(2-(2-(pyrrolidin-1-yl)ethyl)benzoyl)piperidin-2yl)methoxy)benzaldehyde, compound 17.

Compound 17 was synthesized according to Scheme 14.

Scheme 14

1411 Is

Step 1. Synthesis of 2-bromo-3-[[(2S)-1-[2-[(E)-2-ethoxyethenyl]benzyl]piperidin-2-yl]methoxy]phenol.

A solution of [(2S)-1-[2-[(E)-2-ethoxyethenyl]benzoyl]piperidin-2yl]methanol (5.00 g) was placed in a 100 ml round bottom flask, which was purged and maintained under an inert nitrogen atmosphere. , 0.017 mmol, 1.00 eq.) in THF (50 ml), 2-bromobenzene-1,3-diol (3.27 g, 0.017 mmol, 1 eq.), diisopropyl azodicarboxylate (4.19 g, 0.021 mmol, 1.2 eq.) and PPh3 (5.44 g, 0.021 mmol, 1.2 eq.). The resulting solution was stirred for 1 hour at 0°C in an ice/salt bath, then removed from the bath and left to stir overnight at room temperature. The reaction was then quenched by adding water/ice. The resulting solution was extracted with 100 ml of ethyl acetate, dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1:4) as eluent. 2-6pom-3-[[(2S)-1-[2[(E)-2-ethoxyethenyl]benzoyl]piperidin-2-yl]methoxy]phenol was obtained. LCMS (ES) [M+1]+ m/z 460.2.

Step 2. Synthesis of 2-[2-[(2S)-2-(2-bromo-3-hydroxyphenoxymethyl)piperidine-1-carbonyl]phenyl]acetaldehyde.

2-6pom-3-[[(2S)-1-[2-[(E)-2ethoxyethenyl]benzoyl]piperidin-2-yl]methoxy]phenol (3.00 g, 6.517 mmol) was placed in a 100 ml round bottom flask , 1.00 eq.) and 1 M HCl in EtOAc (20 ml). The resulting solution was stirred overnight at room temperature. The resulting mixture was concentrated under vacuum. 2-[2-[(2S)-2-(2-6pom-3hydroxyphenoxymethyl)piperidine-1-carbonyl]phenyl]acetaldehyde was obtained. LCMS (ES) [M+1]+m/z 432.2.

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Step 3. Synthesis of 2-bromo-3-[[(28)-1-[2-[2-(pyrrolidin-1-yl)ethyl]benzoyl]piperidin-2yl]methoxy]phenol.

A solution of 2-[2-[(2S)-2-(2-6pom-3hydroxyphenoxymethyl)piperidine-1-carbonyl]phenyl]acetaldehyde (2.20 g, 0.005 mmol, 1.00 eq.) was placed in a 100 mL round bottom flask. ) in MeOH (10 ml), pyrrolidine (0.54 g, 0.008 mmol, 1.5 eq.), MeOH (1 eq.) and NaBH ₄ (0.48 g, 0.013 mmol, 2.5 eq.). The resulting solution was stirred for 1 hour at 0°C in an ice/salt bath. The resulting solution was stirred overnight at room temperature. The reaction was then quenched by adding water/ice. The resulting solution was extracted with 100 ml of dichloromethane, dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography using dichloromethane/methanol (1:10) as eluent. 2-6pom-3-[[(2S)-1-[2-[2(pyrrolidin-1-yl)ethyl]benzoyl]piperidin-2-yl]methoxy]phenol was obtained. LCMS (ES) [M+1]+ m/z 487.2.

Step 4. Synthesis of 2-ethenyl-3-[[(2S)-1-[2-[2-(pyrrolidin-1-yl)ethyl]benzoyl]piperidin-2yl]methoxy]phenol.

2-bromo-3-[[(2S)-1-[2-[2-(pyrrolidin-1-yl)ethyl]benzoyl]piperidine was placed in a 25 ml round bottom flask, which was purged and maintained inside an inert atmosphere of nitrogen. -2yl]methoxy]phenol (600.00 mg, 1.231 mmol, 1.00 eq.) in dioxane (10 ml), 2-ethenyl-4,4,5,5-tetramethyl1,3,2-dioxaborolane (189, 59 mg, 1.231 mmol, 1 eq.), K2CO3 (340.24 mg, 2.462 mmol, 2 eq.) and Pd(dppf)Cl ₂ (45.03 mg, 0.062 mmol, 0.05 eq.). The resulting solution was stirred overnight at 80°C in an oil bath. The reaction was then quenched by adding water. The resulting solution was extracted with 100 ml of dichloromethane, dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (2:1) as eluent. 2-ethenyl-3-[[(2S)-1-[2-[2-(pyrrolidin-1-yl)ethyl]benzoyl]piperidin-2yl]methoxy]phenol was obtained. LCMS (ES) [M+1]+ m/z 435.1.

Step 5. Synthesis of (S)-2-hydroxy-6-((1-(2-(2-(pyrrolidin-1-yl)ethyl)benzoyl)piperidin-2yl)methoxy)benzaldehyde.

A solution of 2-ethenyl-3-[[(2S)-1-[2-[2-(pyrrolidin-1yl)ethyl]benzoyl]piperidin-2-yl]methoxy]phenol (120.00) was placed in a 10 ml round bottom flask. mg, 0.276 mmol, 1.00 eq.) in acetone (5 ml), a solution of NaIO4 (118.12 mg, 0.552 mmol, 2 eq.) in H2O (2 ml) and K2OSO4-2H2O (10.17 mg, 0.028 mmol, 0.1 eq.). The resulting solution was stirred overnight at room temperature. Solids were filtered off and the filtrate was concentrated. The residue was purified by silica gel column chromatography using acetonitrile:H ₂ O (1:4) as eluent. (S)-2-hydroxy-6-((1(2-(2-(pyrrolidin-1-yl)ethyl)benzoyl)piperidin-2-yl)methoxy)benzaldehyde was obtained. LCMS (ES) [M+1]+ m/z 437.3. 1H NMR (300 MHz, chloroform-d) δ 12.02 (broad s, 1H), 10.46-10.32 (m, 1H), 7.53-7.29 (m, 3H), 7.28 -7.12 (m, 2H), 6.64-6.38 (m, 2H), 5.66-5.21 (m, 1H), 4.49-3.99 (m, 2H), 3 .58-2.36 (m, 10N), 2.16-1.16 (m, 10N).

Example 15. (S)-2-G hydroxy-6-(( 1 -(3-(2-hydroxyethyl)pyrazin-2-carbonyl)pyrrolidin-2yl)methoxy)benzaldehyde, compound 19.

Compound 19 was synthesized according to Scheme 15.

Scheme 15

Step 1. Synthesis of methyl 3-ethenylpyrazine-2-carboxylate.

A 250 mL round bottom flask, purged and maintained under an inert nitrogen atmosphere, was charged with methyl 3-bromopyrazine-2-carboxylate (5.00 g, 23.04 mmol, 1.00 eq), 2-ethenyl-4 ,4,5,5-tetramethyl-1,3,2-dioxaborolane (4.26 g, 27.66 mmol, 1.20 eq.), dioxane (60.00 ml), H ₂ O (10.00 ml ), K ₂ CO ₃ (6.37 g, 46.08 mmol, 2.00 eq.) and Pd(dppf)Cl ₂ (1.69 g, 2.30 mmol, 0.10 eq.). The resulting solution was stirred for 5 hours at 80°C. The reaction mixture was cooled to room temperature. Solids were filtered off. The filtrate was concentrated and the resulting residue was purified by silica gel column chromatography using

THF/petroleum ether (15%) as eluent. Methyl 3-ethenylpyrazine-2-carboxylate was obtained.

LCMS (ES) [M+1]+ m/z: 165.

Stage 2. Synthesis of 3-(2-methoxyethyl)pyrazine-2-carboxylic acid.

A 250 mL round bottom flask was filled with methyl 3-ethenylpyrazine-2-carboxylate (3.50 g, 21.32 mmol, 1.00 eq), MeOH (40.00 mL), and NaOMe (3.46 g, 64 .05 mmol, 3.00 equivalent). The resulting solution was stirred overnight at 70°C. The resulting mixture was concentrated. The pH value of the solution was adjusted to 2-3 using HCl (1 mol/l). The resulting mixture was concentrated. The residue was purified by silica gel column chromatography using MeOH/DCM (10%) as eluent. 3-(2-methoxyethyl)pyrazine-2-carboxylic acid was obtained. LCMS (ES) [M+1] ⁺ m/z: 183.

Step 3. Synthesis of [(2S)-1-[3-(2-methoxyethyl)pyrazin-2-carbonyl]pyrrolidin-2-yl]methanol.

3-(2-methoxyethyl)pyrazine-2carboxylic acid (1.50 g, 8.23 mmol, 1.00 equiv.), prolinol (0.83 g, 8.21 mmol, 1 .00 eq.), N,Ndiisopropylethylamine (3.19 g, 24.70 mmol, 3.00 eq.) and DMF (30.00 ml). After this, 1-[bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide-hexafluorophosphate (3.76 g, 9.88 mmol, 1 .20 eq.) at 0°C. The resulting solution was stirred overnight at room temperature. The crude product was purified by preparative-HPLC under the following conditions: column, XBridge Prep C18 OBD column, 19 cm, 150 mm, 5 µm; mobile phase, water (0.1% HCOOH) and MeCN (5% to 20% phase B for 10 minutes); detector, 254. [(2S)-1-[3-(2methoxyethyl)pyrazin-2-carbonyl]pyrrolidin-2-yl]methanol was obtained.

LCMS (ES) [M+1] ⁺ m/z: 266.

Step 4. Synthesis of 2-hydroxy-6-[[(2S)-1-[3-(2-methoxyethyl)pyrazin-2-carbonyl]pyrrolidin-2yl]methoxy]benzaldehyde.

[(2S)-1-[3-(2-methoxyethyl)pyrazin-2carbonyl]pyrrolidin-2-yl]methanol (1.00 g , 3.77 mmol, 1.00 eq.), 2,6-dihydroxybenzaldehyde (0.62 g, 4.49 mmol, 1.19 eq.), PPh ₃ (1.19 g, 4.52 mmol, 1 .20 eq.) and DCM (30.00 ml). Diisopropyl azodicarboxylate (0.91 g, 4.52 mmol, 1.20 eq) was then added dropwise with stirring at 0°C. The resulting solution was stirred overnight at room temperature. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography using ethyl acetate/DCM (10%) as eluent. 2-hydroxy-6-[[(2S)-1-[3-(2-methoxyethyl)pyrazine-2carbonyl]pyrrolidin-2-yl]methoxy]benzaldehyde was obtained. LCMS (ES, m/z): [M+H]+: 386.2.

Step 5. Synthesis of (S)-2-hydroxy-6-((1-(3-(2-hydroxyethyl)pyrazine-2-carbonyl)pyrrolidin-2yl)methoxy)benzaldehyde.

2-hydroxy-6-[[(2S)-1-[3-(2methoxyethyl)pyrazine-2-carbonyl]pyrrolidin-2-yl]methoxy]benzaldehyde (360.00 mg, 0 .93 mmol, 1.00 eq.) and DCM (10.00 ml). BBg ₃ /DCM (9.34 mL, 9.34 mmol, 10.00 eq) was then added dropwise with stirring at -78°C. The resulting solution was stirred for 2 hours at 0°C. The reaction was then quenched by adding 20 mL of water/ice. The resulting solution was extracted with 3x20 ml dichloromethane, the organic layers were combined, dried over anhydrous sodium sulfate and concentrated. The crude product was purified by preparative-HPLC under the following conditions: column, XBridge Prep C18 OBD column, 19 cm, 150 mm, 5 µm; mobile phase, water (0.1% HCOOH) and acetonitrile (20% to 50% phase B for 11 minutes); detector, 254. (S)-2-hydroxy-6-((1-(3-(2hydroxyethyl)pyrazine-2-carbonyl)pyrrolidin-2-yl)methoxy)benzaldehyde was obtained. LCMS: (ES, m/z): [M+H]+: 372. 1H NMR: (300 MHz, DMSO^) δ 11.77 (s, 1H), 10.34 (s, 1H), 8, 66 (t, J= 2.4 Hz, 1H), 8.51-8.46 (m, 1H), 7.57-7.40 (m, 1H), 6.72 (d, J= 8, 3 Hz, 1H), 6.55-6.33 (m, 1H), 4.61-4.54 (m, 1H), 4.34 (d, J= 4.9 Hz, 2H), 3.933, 63 (m, 3H), 3.38-3.19 (m, 2H), 3.06-2.76 (m, 2H), 2.23-1.87 (m, 3H), 1.84- 1.74 (m, 1H).

Example 16. 2-Hydroxy-6-{[(3S)-4-[2-(2-methoxyethyl)pyridin-3-carbonyl]morpholin-3yl]methoxy}benzaldehyde, compound 20.

Compound 20 was synthesized according to Scheme 16.

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Scheme 16

16a 16b 16c

Stage 1.

Methyl 2-chloropyridine-3-carboxylate (10.0 g, 58.28 mmol, 1.0 eq.), dioxane (100 mL) was placed in a 250 mL round bottom flask, purged and maintained under an inert nitrogen atmosphere. , tributyl(ethenyl) stannane (37.0 g, 116.56 mmol, 2.0 eq.) and Pd(dppf)Cl ₂ (4.26 g, 5.83 mmol, 0.1 eq.). The mixture was stirred for 12 hours at 80°C in an oil bath. After cooling to room temperature, the reaction mixture was concentrated under reduced pressure. The residue was purified on a silica gel column using ethyl acetate/petroleum ether (1:3) as eluent. Methyl 2-ethenylpyridine-3-carboxylate was obtained. LCMS (ES) [M+1]+ m/z: 164.

Stage 2.

Methyl 2-ethenylpyridine-3-carboxylate (7.80 g, 47.90 mmol, 1.0 eq) and MeOH (50 ml), HCl (c) (8.0 ml) were placed in a 250 ml round bottom flask. . The reaction solution was stirred for 12 hours at 90°C in an oil bath. The reaction mixture was cooled to room temperature. Methyl 2-(2-methoxyethyl)pyridine-3-carboxylate hydrochloride was obtained and used directly in the next step without further purification. LCMS (ES) [M-HC1+1] ⁺ m/z: 196.

Stage 3.

Methyl 2-(2-methoxyethyl)pyridine-3-carboxylate hydrochloride (7.0 g, 30.30 mmol, 1.0 eq.), MeOH/H ₂ O (1:2) was placed in a 250 ml round bottom flask. (150 ml) and NaOH (2.40 g, 60.60 mmol, 2.0 eq.). The mixture was stirred for 2 hours at 50°C in an oil bath. After cooling to room temperature, the solution was concentrated under reduced pressure. The pH of the residue was adjusted to 6 with (6 M) HCl and purified on a C18-120 g column under conditions: CH ₃ CN/H ₂ O increased from 5% to 60% over 12 minutes. 2-(2-methoxyethyl)pyridine-3-carboxylic acid hydrochloride was obtained. LCMS (ES) [M-HCl+1]+ m/z: 182.

Stage 4.

2-(2-methoxyethyl)pyridine-3-carboxylic acid hydrochloride (1.0 g, 4.60 mmol, 1.0 eq.) was placed in a 50 mL three-neck round bottom flask, purged and maintained under an inert nitrogen atmosphere. , DMF (20 ml), N,N-diisopropylethylamine (2.38 g, 18.40 mmol, 4.0 eq.) and (3R)-morpholin-3-ylmethanol hydrochloride (0.85 g, 5.51 mmol , 1.2 eq.). After this, 1-[bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxidehexafluorophosphate (2.10 g, 5.51 mmol, 1. 2 equiv.) at 0°C. The reaction solution was stirred for 12 hours at room temperature. The reaction solution was diluted with 30 ml H ₂ O and extracted with 3x100 ml ethyl acetate. The combined organic phase was washed with 3x50 ml brine and dried over anhydrous sodium sulfate. The solution was filtered and the filtrate was concentrated under reduced pressure. The residue was purified on a silica gel column with ethyl acetate/petroleum ether (1:1). [(3C)-4-[2-(2-methoxyethyl)pyridin-3-carbonyl]morpholin-3-yl]methanol was obtained. LCMS (ES) [M+1]+ m/z: 281.

Stage 5.

[(2S)-1-[2-(2-methoxyethyl)pyridin-3-carbonyl]pyrrolidin-2yl]methanol (280 mg, 1. 06 mmol, 1.0 eq.), THF (10 ml), 2,6-dihydroxybenzaldehyde (176 mg, 1.27 mmol, 1.2 eq.) and PPh ₃ (333 mg, 1.27 mmol, 1. 2 eq.). The mixture was cooled to 0° C. followed by the dropwise addition of a solution of di-tert-butyl azodicarboxylate (293 mg, 1.27 mmol, 1.2 eq.) in THF (2 ml) with stirring. After addition, the reaction solution was stirred for 12 hours at room temperature. The resulting mixture was concentrated in vacuo to remove the solvent, and the crude product was purified by preparative-HPLC under the following conditions (IntelFlash-1):

- 66 045055 column: Ascentis Express C18, 50x3.0 mm, 2.7 µm, mobile phase A: water/0.05% formic acid, mobile phase B: CH ₃ CN, flow rate: 1.5 ml/min, gradient: 5% to 100% B over 1.2 minutes, hold 0.6 minutes. As a result, 2-hydroxy-6-[[(2S)-1-[2-(2-methoxyethyl)pyridin-3carbonyl]pyrrolidin-2-yl]methoxy]benzaldehyde was isolated. LCMS: (ES, m/z): [M+H]+: 401.2. 1H NMR: (300 MHz, DMSO-d ₆ , ppm): δ 11.76 (s, 1H), 10.12 (s, 1H), 8.57 (dd, J= 4.8, 1 .8 Hz, 1H), 7.73-7.31 (m, 3H), 6.75 (d, J= 8.4 Hz, 1H), 6.56 (d, J= 8.1 Hz, 1H ), 5.05-4.89 (m, 1H), 4.45-4.33 (m, 2H), 4.11-3.92 (m, 1H), 3.73-3.35 (m , 6H), 3.20-2.79 (m, 6H).

Example 17. 2-Hydroxy-6-{[(3S)-4-[3-(2-hydroxyethyl)pyrazin-2-carbonyl]morpholin-3yl]methoxy}benzaldehyde, compound 21.

Compound 21 was synthesized according to Scheme 17.

Scheme 17

2-hydroxy-6-[[(3S)-4-[3-(2methoxyethyl)pyrazin-2-carbonyl]morpholin-3-yl]methoxy]benzaldehyde (500.00 mg, 1 .25 mmol, 1.00 eq.), which was prepared as described in Scheme 23, and DCM (10.00 ml). BBg ₃ /DCM (12.46 mL, 12.46 mmol, 10.00 eq) was then added dropwise with stirring at -78°C. The resulting solution was stirred for 2 hours at 0°C. The resulting solution was extracted with 3x20 ml dichloromethane, the organic layers were combined, dried over anhydrous sodium sulfate and concentrated. The crude product was purified by preparative-HPLC under the following conditions: column, XBridge Prep C18 OBD column, 19 cm, 150 mm, 5 µm; mobile phase, water (0.1% HCOOH) and acetonitrile (20% to 50% phase B for 11 minutes); detector, 254. 2-Hydroxy-6-[[(3S)-4-[3-(2hydroxyethyl)pyrazin-2-carbonyl]morpholin-3-yl]methoxy]benzaldehyde was obtained. LCMS: (ES, m/z): [M+H]+: 388. 1H NMR: (300 MHz, DMSOL) δ 11.77 (d, J= 11.9 Hz, 1H), 10.26 (d , J = 13.1 Hz, 1H), 8.68-8.66 (m, 1H), 8.49-8.45 (m, 1H), 7.57 (d, J = 8.4 Hz, 1H), 7.56-7.43 (m, 1H), 6.75-6.49 (m, 2H), 4.99-4.93 (m, 1H), 4.76-4.63 ( m, 1H), 4.55-4.32 (m, 2H), 4.14-3.89 (m, 1H), 3.86-3.68 (m, 2H), 3.73-3. 61 (m, 1H), 3.66-3.46 (m, 1H), 3.513.30 (m, 2H), 3.17-2.80 (m, 3H).

Example 18. 2-Hydroxy-6-{[(3S)-4-[2-(2-hydroxyethyl)benzoyl]morpholin-3-yl]methoxy}benzaldehyde, compound 22.

Compound 22 was synthesized according to Scheme 18.

Scheme 18

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Stage 1.

Place methyl 2-bromobenzoate (5.00 g, 23.251 mmol, 1.00 equiv.), dioxane (60.00 mL), water (10 mL), 2-[(E)-2 into a 250 mL round bottom flask. -ethoxyethenyl]-4,4,5,5-tetramethyl-1,3,2dioxaborolane (6.91 g, 34.876 mmol, 1.50 eq.), sodium methaneperoxoate (4.98 g, 46.502 mmol, 2.00 eq. .) and tetrakis(triphenylphosphine)palladium(0) (2.69 g, 2.325 mmol, 0.10 eq.). The resulting solution was stirred for 16 hours at 80°C. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1:3) as eluent. The collected fractions were combined and concentrated. Methyl 2-[(E)-2ethoxyethenyl]benzoate was obtained. LCMS (ES) [M+1]+ m/z 207.1.

Stage 2.

A 100 mL round bottom flask was filled with methyl 2-[(E)-2-ethoxyethenyl]benzoate (2.40 g, 11.637 mmol, 1.00 eq), methanol (30.00 mL), and Pd/C (240 .00 mg). The flask was evacuated and purged with nitrogen three times, followed by hydrogen. The mixture was stirred for 2 hours at room temperature under a hydrogen atmosphere. Solids were filtered off. The resulting filtrate was concentrated to give methyl 2-(2-ethoxyethyl)benzoate. LCMS (ES) [M+1] ⁺ m/z 209.1.

Stage 3.

A 50 mL round bottom flask was filled with methyl 2-(2-ethoxyethyl) benzoate (2.00 g, 9.604 mmol, 1.00 eq.), methanol (10.00 mL), water (10.00 mL), caustic solution. soda (0.77 g, 19.251 mmol, 2.00 eq.). The resulting solution was stirred for 4 hours at 25°C. The resulting solution was diluted with 50 ml of water. The pH of the solution was adjusted to 5 using HCl (1 mol/L). The solids were collected by filtration to give 2-(2-ethoxyethyl)benzoic acid. LCMS (ES) [M+1] ⁺ m/z 195.1.

Stage 4.

Place 2-(2-ethoxyethyl)benzoic acid (1.50 g, 7.723 mmol, 1.00 eq), DCM (30.00 mL), (3R)-morpholin-3-ylmethanol ( 0.90 g, 7.723 mmol, 1.00 eq.), 1[bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide-hexafluorophosphate (4 .40 g, 11.584 mmol, 1.50 eq.) and K,K-diisopropylethylamine (2.99 g, 23.168 mmol, 3.00 eq.). The resulting solution was stirred for 2 hours at 25°C. The resulting mixture was concentrated. The residue was purified on a silica gel column using ethyl acetate/petroleum ether (1:1) as eluent. The collected fractions were combined and concentrated to give [(3R)-4-[2-(2-ethoxyethyl)benzoyl]morpholin-3yl]methanol. LCMS (ES) [M+1] ⁺ m/z 294.2.

Stage 5.

[(3R)-4-[2-(2-ethoxyethyl)benzoyl]morpholin-3yl]methanol (600.00 mg, 2.045 mmol, 1.00 eq.), tetrahydrofuran (20 mL) was placed in a 520 mL round bottom flask. , 2,6-dihydroxybenzaldehyde (282.49 mg, 2.045 mmol, 1.00 eq.), triphenylphosphine (643.74 mg, 2.454 mmol, 1.20 eq.) and diisopropyl azodicarboxylate (496.28 mg, 2.454 mmol, 1 ,20 eq.). The resulting solution was stirred for 16 hours at 25°C. The resulting mixture was concentrated. The residue was purified on a silica gel column using ethyl acetate/petroleum ether (1:2) as eluent. The collected fractions were combined and concentrated. 2-[[(3S)-4-[2-(2-ethoxyethyl)benzoyl]morpholin-3-yl]methoxy]-6hydroxybenzaldehyde was obtained. LCMS (ES) [M+1] ⁺ m/z 414.2.

Stage 6.

2-[[(3S)-4-[2-(2-ethoxyethyl)benzoyl]morpholin-3yl]methoxy]-6-hydroxybenzaldehyde (200 mg, 0.48 mmol, 1.0 equiv) was placed in a 50 ml round bottom flask .), DXM (20 ml). Boron tribromide (2.4 mL, 2.4 mmol, 5.0 eq, 1 M) was then added dropwise at -78°C. The resulting solution was stirred for 3 hours at 0°C. The reaction was then quenched by adding 10 ml of water. The resulting solution was extracted with 2x20 ml dichloromethane and concentrated. The crude reaction mixture was filtered and subjected to reverse phase preparative HPLC (Prep-C18, 20-45M, 120 g, Tianjin Bonna-Agela Technologies; gradient elution from 25% MeCN in water to 35% MeCN in water over a 10-minute period, where both solvent contained 0.1% formic acid) to obtain 2hydroxy-6-{[(3S)-4-[2-(2-hydroxyethyl)benzoyl]morpholin-3-yl]methoxy}benzaldehyde. LCMS (ES) [M+1] ⁺ m/z 386.1. ^1H NMR (300 MHz, DMSO^) δ 11.76 (broad s, 1H), 10.29 (s, 1H), 7.62-7.50 (m, 1H), 7.48-6.95 (m, 4H), 6.88-6.48 (m, 2H), 5.00-4.20 (m, 4H), 4.17-3.41 (m, 7H), 3.23-29 .5 (m, 1H), 2.94-2.57 (m, 2H).

Example 19. 2-Hydroxy-6-{[(3S)-4-[2-(hydroxymethyl)benzoyl]morpholin-3-yl]methoxy}benzaldehyde, compound 23.

Compound 23 was synthesized according to scheme 19.

- 68 045055

Scheme 19

SIEILSH ON

19e 23

Stage 1.

A 500 mL round bottom flask was filled with phthalide (11.00 g, 82.008 mmol, 1.00 eq.), H2O (200.00 mL, 11101.675 mmol, 135.37 eq.), and NaOH (4.92 g, 123.009 mmol, 1.50 eq.). The resulting solution was stirred overnight at 100°C in an oil bath. The reaction mixture was cooled to 0°C in a water/ice bath. The pH of the solution was adjusted to 5 using HCl (6 mol/L). Solids were collected by filtration. The solid was dried in an oven. 2-hydroxymethylbenzoic acid was obtained. LCMS (ES) [M-1] ^- m/z 151.1.

Stage 2.

Place 2-hydroxymethylbenzoic acid (5.00 g, 32.863 mmol, 1.00 eq), DCM (100.00 mL) and imidazole (4.47 g, 65.725 mmol, 2.00 eq) into a 250 mL round bottom flask. ). Thereafter, tert-butyl(chloro)diphenylsilane (10.84 g, 39.435 mmol, 1.20 eq.) was added in several portions at 0° C. over 5 minutes. The resulting solution was stirred for 2 hours at room temperature. The resulting solution was diluted with 60 ml of water. The resulting solution was extracted with 3x100 ml dichloromethane, and the organic layer was washed with 1x60 ml brine, dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography using dichloromethane/ethyl acetate (10% ethyl acetate-20% ethyl acetate). 2-[[(tert-butyldiphenylsilyl)oxy]methyl]benzoic acid was obtained. LCMS (ES) [M+1]+ m/z 391.2.

Stage 3.

2-[[(tert-butyldiphenylsilyl)oxy]methyl]benzoic acid (2.00 g, 5.121 mmol, 1.00 eq), DCM was placed in a 250 mL round bottom flask, purged and maintained under an inert nitrogen atmosphere. (120 ml) and oxalyl chloride (1.30 g, 10.242 mmol, 2.00 eq.). The resulting solution was stirred for 4 hours at room temperature. The resulting mixture was concentrated under vacuum. (3R)-morpholine-3ylmethanol hydrochloride (0.94 g, 6.145 mmol, 1.20 eq) and triethylamine (1.55 g) were placed in another 250 mL three-neck round bottom flask, purged and maintained under an inert nitrogen atmosphere. , 15.318 mmol, 2.99 eq.). A solution of 2-[[(tert-butyldiphenylsilyl)oxy]methyl]benzoyl chloride (2.00 g, 4.890 mmol, 1.00 eq.) in DCM (30 mL) was then added dropwise with stirring at 0° C. over 30 minutes. The resulting solution was stirred for 2 hours at room temperature. The resulting solution was diluted with 200 ml of DCM. The resulting mixture was washed with 1x100 ml 1 M HCl. The mixture was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (50% ethyl acetate). [(3R)-4-(2-[[(tert-butyldiphenylsilyl)oxy]methyl]benzoyl)morpholin-3-yl]methanol was obtained. LCMS (ES) [M+1]+ m/z 490.3.

Stage 4.

[(3R)-4-(2-[[(tertbutyldiphenylsilyl)oxy]methyl]benzoyl)morpholin-3-yl]methanol (1. 00 g, 2.042 mmol, 1.00 eq.), 2,6-dihydroxybenzaldehyde (0.56 g, 4.084 mmol, 2.00 eq.), PPh ₃ (1.07 g, 4.084 mmol, 2.00 eq. ) and THF (60.00 ml). The resulting solution was stirred for 15 minutes at 0°C. Diisopropyl azodicarboxylate (825.87 mg, 4.084 mmol, 2.00 eq) was then added dropwise with stirring at 0°C for 5 minutes. The resulting solution was stirred for 5 hours at room temperature. The resulting solution was diluted with 30 ml of water. The resulting solution was extracted with 3x60 ml dichloromethane; the organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by petroleum ether/THF (10% THF) silica gel column chromatography. Received 2- 69 045055

[[(3S)-4-(2-[[(tert-butyldiphenylsilyl)oxy]methyl]benzoyl)morpholin-3-yl]methoxy]-6-hydroxybenzaldehyde. LCMS (ES) [M+1]+ m/z 610.3.

Stage 5.

2-[[(3B)-4-(2-[[(tertbutyldiphenylsilyl)oxy]methyl]benzoyl)morpholin-3-yl]methoxy]-6-hydroxybenzaldehyde (0.80 g, 1.312) was placed in a 100 ml round bottom flask. mmol, 1.00 equiv.), THF (30.00 ml) and tetrabutylammonium fluoride (1.32 ml, 2.0 equiv., 2 M). The resulting solution was stirred for 2 hours at room temperature. The resulting solution was diluted with 10 ml of water. The resulting solution was extracted with 3x60 ml dichloromethane, the organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified on a silica gel column using petroleum ether/THF (55% THF) as eluent. The crude product was further purified by flash preparative-HPLC (Prep-C18, 20-45M, 120 g, Tianjin Bonna-Agela Technologies; gradient elution from 35% MeCN in water to 60% MeCN in water over a 10-minute period, where both solvents contained 0.1% formic acid). 2-hydroxy-6-[[(3S)-4-[2(hydroxymethyl)benzoyl]morpholin-3-yl]methoxy]benzaldehyde was obtained. LCMS (ES) [M+1]+ m/z 372.1. 1H NMR (300 MHz, DMSO-d6, ppm) δ 11.78 (s, 1H), 10.21 (s, 1H), 7.70-7.21 (m, 5H), 6.77 (d, J= 8.2 Hz, 1H), 6.56 (d, J= 8.4 Hz, 1H), 5.32-5.15 (m, 1H), 5.03-4.21 ( m, 5H), 4.18-3.84 (m, 2H), 3.78-3.55 (m, 2H), 3.42-3.36 (m, 1H), 3.11-2, 98 (m, 1H).

Example 20. 2-Hydroxy-6-{[(2S)-1-[2-(2-methoxyethyl)pyridin-3-carbonyl]pyrrolidin-2yl]methoxy}benzaldehyde, compound 24.

Compound 24 was synthesized according to scheme 20.

Scheme 20

Stage 1.

2-(2-methoxyethyl)pyridine-3-carboxylic acid (1.0 g, 5.52 mmol, 1.0 eq.) was placed in a 50 mL three-neck round bottom flask, purged and maintained under an inert nitrogen atmosphere. DMF (20 ml), prolinol (670 mg, 6.62 mmol, 1.2 eq.) and N,Ndiisopropylethylamine (2.85 g, 22.08 mmol, 4.0 eq.). After this, 1[bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide-hexafluorophosphate (2.52 g, 6.62 mmol, 1 .2 eq.) at 0°C. After addition, the mixture was stirred for 12 hours at room temperature. The reaction solution was diluted with 30 ml H2O and extracted with 3x100 ml ethyl acetate. The combined organic phase was washed with 3x50 ml brine and dried over anhydrous sodium sulfate. The solution was filtered and the filtrate was concentrated under reduced pressure. The residue was purified on a silica gel column with ethyl acetate. [(2S)-1-[2-(2-methoxyethyl)pyridin-3carbonyl]pyrrolidin-2-yl]methanol was obtained.

LCMS (ES) [M+1]+ m/z: 265.

Stage 2.

[(2S)-1-[2-(2-methoxyethyl)pyridin-3-carbonyl]pyrrolidin-2yl]methanol (380 mg, 1. 44 mmol, 1.0 eq.), THF (20 ml), 2,6-dihydroxybenzaldehyde (199 mg, 1.44 mmol, 1.0 eq.) and PPh ₃ (377 mg, 1.44 mmol, 1.0 eq.) 0 eq.). The mixture was cooled to 0°C and stirred for 15 minutes. A solution of di-tert-butyl azodicarboxylate (331 mg, 1.44 mmol, 1.0 eq) in THF (2 ml) was then added dropwise with stirring. After addition, the reaction solution was stirred for 12 hours at room temperature. The solution was then concentrated under reduced pressure to remove the solvent. The residue was purified by flash preparative-HPLC under the following conditions (IntelFlash-1): column: Ascentis Express C18, 50x3.0 mm, 2.7 µm, mobile phase A: water/0.05% formic acid, mobile phase B : CH ₃ CN, flow rate: 1.5 ml/min, gradient: 5% to 100% B over 1.2 minutes, hold 0.6 minutes. 2-hydroxy-6-{[(2S)-1-[2-(2methoxyethyl)pyridin-3-carbonyl]pyrrolidin-2-yl]methoxy}benzaldehyde was obtained. LCMS: (ES, m/z): [M+H]+: 385. 1H NMR: (300 MHz, DMSOL, ppm): δ 11.78 (s, 1H), 10.33 (s, 1H), 8.55. (dd, J= 1.5, 4.8 Hz, 1H), 7.64 (dd, J= 7.5, 1.8 Hz, 1H), 7.57-7.52 (m, 1H), 7.32-7.28 (m, 1H), 6.70 (d, J= 8.4 Hz, 1H), 6.53 (d, J= 8.4 Hz, 1H), 4.57-4 .53 (m, 1H), 4.44-4.39 (m, 1H), 4.31-4.26 (m, 1H), 3.79-3.52 (m, 2H), 3.31 -3.11 (m, 2H), 3.11 (s, 3H), 3.04-2.79 (m, 2H), 2.24-1.89 (m, 3H), 1.84-1 .76 (m, 1H).

- 70 045055

Example 21. 2-G hydroxy-6-{[(2S)-1-[3-(2-methoxyethyl)pyrazin-2-carbonyl]pyrrolidin-2yl]methoxy}benzaldehyde, compound 25.

Compound 25 was synthesized according to Scheme 21.

Scheme 21

21a 21b 21c

Stage 1.

A 250 mL round bottom flask, purged and maintained under an inert nitrogen atmosphere, was charged with methyl 3-bromopyrazine-2-carboxylate (5.00 g, 23.04 mmol, 1.00 eq), 2-ethenyl-4 ,4,5,5-tetramethyl-1,3,2-dioxaborolane (4.26 g, 27.66 mmol, 1.20 eq.), dioxane (60.00 ml), H2O (10.00 ml), K2CO3 (6.37 g, 46.08 mmol, 2.00 eq) and Pd(dppf)Cl2 (1.69 g, 2.30 mmol, 0.10 eq). The resulting solution was stirred for 5 hours at 80°C. The reaction mixture was cooled to room temperature. Solids were filtered off. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography using THF/petroleum ether (15%) as eluent. Methyl 3-ethenylpyrazine-2-carboxylate was obtained. LCMS (ES) [M+1]+ m/z: 165.

Stage 2.

A 250 mL round bottom flask was filled with methyl 3-ethenylpyrazine-2-carboxylate (3.50 g, 21.32 mmol, 1.00 eq), MeOH (40.00 mL), and NaOMe (3.46 g, 64 .05 mmol, 3.00 eq.). The resulting solution was stirred overnight at 70°C. The resulting mixture was concentrated. The pH value of the solution was adjusted to 2-3 using HCl (1 mol/l). The resulting mixture was concentrated. The residue was purified on a silica gel column using MeOH/DCM (10%) as eluent. 3-(2methoxyethyl)pyrazine-2-carboxylic acid was obtained. LCMS (ES) [M+1]+ m/z: 183.

Stage 3.

3-(2-methoxyethyl)pyrazine-2carboxylic acid (1.50 g, 8.23 mmol, 1.00 equiv.), prolinol (0.83 g, 8.21 mmol, 1 .00 eq.), N,Ndiisopropylethylamine (3.19 g, 24.70 mmol, 3.00 eq.) and DMF (30.00 ml). After this, 1-[bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide-hexafluorophosphate (3.76 g, 9.88 mmol, 1 .20 eq.) at 0°C. The resulting solution was stirred overnight at room temperature. The crude product was purified by preparative-HPLC under the following conditions: column, XBridge Prep C18 OBD column, 19 cm, 150 mm, 5 µm; mobile phase, water (0.1% HCOOH) and MeCN (5% to 20% phase B for 10 minutes); detector, 254. [(2S)-1-[3-(2methoxyethyl)pyrazin-2-carbonyl]pyrrolidin-2-yl]methanol was obtained.

LCMS (ES) [M+1]+ m/z: 266.

Stage 4.

[(2S)-1-[3-(2-methoxyethyl)pyrazin-2-carbonyl]pyrrolidin-2-yl]methanol (1. 00 g, 3.77 mmol, 1.00 eq.), 2,6-dihydroxybenzaldehyde (0.62 g, 4.49 mmol, 1.19 eq.), PPh ₃ (1.19 g, 4.52 mmol , 1.20 eq.) and DCM (30.00 ml). Diisopropyl azodicarboxylate (0.91 g, 4.52 mmol, 1.20 eq) was then added dropwise with stirring at 0°C. The resulting solution was stirred overnight at room temperature. The resulting mixture was concentrated. The residue was applied to a silica gel column with ethyl acetate/DCM (10%). 2-hydroxy-6-[[(2S)-1-[3-(2-methoxyethyl)pyrazine-2-carbonyl]pyrrolidin-2yl]methoxy]benzaldehyde was obtained. LCMS: (ES, m/z): [M+H]+: 386.2. 1H NMR: (300 MHz, DMSO-d ₆ ) δ δ 11.77 (s, 1H), 10.34 (s, 1H), 8.67 (d, J= 2.6 Hz, 1H), 8, 51 (d, J = 2.5 Hz, 1H), 7.57-7.40 (m, 1H), 6.72-6.33 (m, 2H), 4.58-4.24 (m, 3H), 3.83-3.43 (m, 2H), 3.39-3.19 (m, 2H), 3.13 (s, 3H), 3.10-2.86 (m, 2H) , 2.23-1.73 (m, 4H).

- 71 045055

Example 22. 2-Hydroxy-6-{[(28)-1-[2-(hydroxymethyl)benzoyl]pyrrolidin-2-yl]methoxy}benzaldehyde, compound 26.

Compound 26 was synthesized according to Scheme 22.

Scheme 22

22a 22c

22d ²⁶

Stage 1.

2-[[(tert-butyldiphenylsilyl)oxy]methyl]benzoic acid (2.00 g, 5.121 mmol, 1.00 eq), DCM was placed in a 100 mL round bottom flask, purged and maintained under an inert nitrogen atmosphere. (60.00 ml) and DMF (0.05 ml, 0.646 mmol, 0.13 eq.). Oxalyl chloride (1.30 g, 10.243 mmol, 2.00 eq.) was then added. The resulting solution was stirred overnight at room temperature. The resulting mixture was concentrated. 2-[[(tert-butyldiphenylsilyl)oxy]methyl]benzoyl chloride was obtained.

Stage 2.

Prolinol (0.59 g, 5.868 mmol, 1.2 eq.), triethylamine (1.48 g, 14.670 mmol, 3 eq.) and DXM (100.00 ml). A solution of 2-[[(tert-butyldiphenylsilyl)oxy]methyl]benzoyl chloride (2.00 g, 4.890 mmol, 1.00 eq.) in DCM (30 mL) was then added dropwise with stirring at 0°C for 30 minutes. . The resulting solution was stirred for 2 hours at room temperature. The resulting solution was diluted with 100 ml of DCM. The resulting mixture was washed with 1x70 ml 1 M HCl. The mixture was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (55% ethyl acetate) as eluent. [(2S)-1-(2-[[(tert-butyldiphenylsilyl)oxy]methyl]benzoyl)pyrrolidin-2yl]methanol was obtained. LCMS (ES) [M+1]+ m/z 474.2.

Stage 3.

[(2S)-1-(2-[[(tertbutyldiphenylsilyl)oxy]methyl]benzoyl)pyrrolidin-2-yl]methanol (1. 00 g, 2.111 mmol, 1.00 eq.), 2,6dihydroxybenzaldehyde (0.58 g, 4.222 mmol, 2.00 eq.), PPh ₃ (1.11 g, 4.222 mmol, 2.00 eq.) and THF (60 ml). The resulting solution was stirred for 15 minutes at 0°C. Diisopropyl azodicarboxylate (0.85 g, 4.222 mmol, 2.00 eq.) was then added dropwise with stirring at 0° C. over 5 minutes. The resulting solution was stirred for 5 hours at room temperature. The resulting solution was diluted with 30 ml of water. The resulting solution was extracted with 3x100 ml dichloromethane; the organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography using petroleum ether/THF (12% THF) as eluent. 2-[[(2S)-1-(2-[[(tert-butyldiphenylsilyl)oxy]methyl]benzoyl)pyrrolidin2-yl]methoxy]-6-hydroxybenzaldehyde was obtained. LCMS (ES) [M+1]+ m/z 594.3.

Stage 4.

2-[[(2S)-1-(2-[[(tert-butyldifenylsilyl)oxy]methyl]benzoyl)pyrrolidin-2-yl]methoxy]-6-hydroxybenzaldehyde (1.10) was placed in a 100 ml round bottom flask g, 1.852 mmol, 1.00 eq.), THF (30.00 ml) and tetrabutylammonium fluoride (1.9 ml, 2 M). The resulting solution was stirred for 2 hours at room temperature. The resulting solution was diluted with 10 ml of water. The resulting solution was extracted with 3x60 ml dichloromethane; the organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was applied to a petroleum ether/THF (52% THF) silica gel column. The crude product was purified by flash preparative-HPLC (PrepC18, 20-45M, 120 g, Tianjin Bonna-Agela Technologies; gradient elution from 35% MeCN in water to 60% MeCN in water over a 10-minute period, both solvent contained 0.1% formic acid). 2-hydroxy-6-[[(28)-1-[2-(hydroxymethyl)benzoyl]pyrrolidin-2-72 045055 yl]methoxy]benzaldehyde was obtained. LCMS (ES) [M+1]+ m/z 356.1. 1H NMR (300 MHz, DMSO-d ₆ , ppm) δ 11.72 (s,

1H), 10.22 (s, 1H), 7.80-7.13 (m, 5H), 6.78-6.41 (m, 2H), 5.22-5.10 (m, 1H) , 4.58-4.28 (m, 4H), 4.07-3.46 (m,

1H), 3.38-3.09 (m, 6.8 Hz, 2H), 2.18-1.70 (m, 4H).

Example 23. 2-G hydroxy-6-{[(3S)-4-[3-(2-methoxyethyl)pyrazin-2-carbonyl]morpholin-3 yl]methoxy}benzaldehyde, compound 27.

Compound 27 was synthesized according to Scheme 23.

Scheme 23

Stage 1.

Place 3-(2-methoxyethyl)pyrazine-2carboxylic acid (2.00 g, 10.98 mmol, 1.00 eq), (3S)-3-[[(tertbutyldimethylsilyl)oxy] into a 100 mL three-neck round bottom flask. methyl]morpholine (2.54 g, 10.98 mmol, 1.00 eq.), Et ₃ N (2.22 g, 21.94 mmol, 2.00 eq.), DCM (30 ml) and 1- ethyl 3-3-dimethylaminopropylcarbodiimide (2.53 g, 13.17 mmol, 1.20 eq.). Hydroxybenzotriazole (1.78 g, 13.17 mmol, 1.20 eq.) was then added portionwise at 0°C. The resulting solution was stirred overnight at room temperature. The reaction was then quenched by adding 30 ml of water. The resulting solution was extracted with 3x30 ml dichloromethane, the organic layers were combined, dried over anhydrous sodium sulfate and concentrated. The residue was purified on a silica gel column using THF/petroleum ether (40%) as eluent. (3B)-3-[[(tertbutyldimethylsilyl)oxy]methyl]-4-[3-(2-methoxyethyl)pyrazin-2-carbonyl]morpholine was obtained. LCMS (ES) [M+1]+ m/z: 396.

Stage 2.

(3S)-3-[[(tert-Butyldimethylsilyl)oxy]methyl]4-[3-(2-methoxyethyl)pyrazin-2-carbonyl]morpholine (4 g, 10.11 mmol, 1.00 eq.) and ethyl acetate (20.00 ml). To the above, HCl (gas) in ethyl acetate (10.11 mL, 20.22 mmol, 2.00 eq.) was added dropwise with stirring at 0°C. The resulting solution was stirred for 2 hours at room temperature. The resulting mixture was concentrated. The pH value of the solution was adjusted to 7-8 using a saturated NaHCO ₃ solution. The resulting solution was extracted with 5x30 ml dichloromethane, the organic layers were combined, dried over anhydrous sodium sulfate and concentrated. The residue was purified on a silica gel column using dichloromethane/methanol (100/3) as eluent. [(3R)-4[3-(2-methoxyethyl)pyrazin-2-carbonyl]morpholin-3-yl]methanol was obtained. LCMS (ES) [M+1]+ m/z: 282.

Stage 3.

[(3R)-4-[3-(2-methoxyethyl)pyrazin-2carbonyl]morpholin-3-yl]methanol (400.00 mg, 1.42 mmol, 1.00 eq.) was placed into a 100 mL three-neck round bottom flask. ), 2,6-dihydroxybenzaldehyde (235.68 mg, 1.71 mmol, 1.20 eq.), DCM (10.00 ml) and PPh ₃ (447.54 mg, 1.71 mmol, 1.20 eq. .). Diisopropyl azodicarboxylate (345.03 mg, 1.71 mmol, 1.20 eq) was then added dropwise with stirring at 0°C. The resulting solution was stirred overnight at room temperature. The resulting mixture was concentrated. The crude product was purified by preparative-HPLC under the following conditions: column, XBridge Prep C18 OBD column, 19 cm, 150 mm, 5 µm; mobile phase, water (0.1% HCOOH) and acetonitrile (30% to 50% phase B for 11 minutes); detector, 254. 2-Hydroxy-6-{[(3S)-4-[3-(2-methoxyethyl)pyrazin-2carbonyl]morpholin-3-yl]methoxy}benzaldehyde was obtained. LCMS: (ES, m/z): [M+H]+: 402. 1H NMR: (300 MHz, DMSO-46) δ 11.79 (s, 1H), 10.31-10.20 (m, 1H), 8.69-8.66 (m, 1H), 8.52-8.47 (m, 1H), 7.58-7.45 (m, 1H), 6.79-6.70 ( m, 1H), 6.646.48 (m, 1H), 5.00-4.94 (m, 1H)), 4.52-4.31 (m, 2H), 4.14-3.92 (m , 1H), 3.81-3.23 (m, 6H), 3.14 (s, 3H), 3.09-2.88 (m, 3H).

Example 24 3-{3-[(38)-3-[(2-Formyl-3-hydroxyphenoxy)methyl]morpholine-4-carbonyl]pyridin-2yl}propanenitrile, compound 28.

Compound 28 was synthesized according to Scheme 24.

- 73 045055

Scheme 24

Stage 1.

A 40 ml vial was filled with methyl 2-chloropyridine-3-carboxylate (2.00 g, 11.66 mmol, 1.00 equiv.), DMF (15.00 ml), NaOAc (1.91 g, 23. 31 mmol, 2.00 eq.), PPh ₃ (1.22 g, 4.66 mmol, 0.40 eq.), Pd(OAc)2 (0.26 g, 1.17 mmol, 0.10 eq. .)acrylonitrile (3.09 g, 58.28 mmol, 5.00 eq.). The resulting solution was stirred overnight at 130°C in an oil bath. The reaction was then quenched with water (20 ml) and extracted with ethyl acetate (40 ml). The organic layer was washed twice with water (40 ml), dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (0-9.9%) as eluent. Methyl 2-[(1E)-2-cyanoeth-1-en-1-yl]pyridin-3-carboxylate was obtained. LCMS: (ES, m/z): [M+H]+ 189.1.

Stage 2.

A 50 mL round bottom flask was filled with methyl (E)-2-(2-cyanovinyl)nicotinate (1.40 g, 7.44 mmol, 1.00 eq.), CH3OH (20 mL), and Pd/C (140 .0 mg, 10%). _H2 was added to the above. The resulting solution was stirred for 1 hour at room temperature. The solids were filtered off and the mixture was concentrated and purified by silica gel column chromatography using ethyl acetate/petroleum ether (0-15%) as eluent. Methyl 2-(2cyanoethyl)nicotinate was obtained. LCMS: (ES, m/z): [M+H]+: 190.1.

Stage 3.

Methyl 2-(2-cyanoethyl)nicotinate (1.00 g, 5.26 mmol, 1.00 eq) and THF (12 mL) were placed in a 50 mL round bottom flask. A solution of LiOH (0.44 g, 10.49 mmol, 1.99 eq) in H2O (6 mL) was then added. The resulting solution was stirred for 1 hour at room temperature and concentrated. The pH value of the solution was adjusted to 5-6 using HCl (2 mol/l). The residue was purified under the following conditions: column C18; mobile phase, water (0.05% formic acid) and CH3CN (5% to 80% for 8 minutes); detector, 220 and 254 nm; flow rate 40 ml/min. 2-(2cyanoethyl)nicotinic acid was obtained. LCMS (ES, m/z): [M+H]+: 177.1.

Stage 4.

Place 2-(2-cyanoethyl)-pyridine-3-carboxylic acid (300.0 mg, 1.70 mmol, 1.00 eq.), 1-[bis-(dimethylamino)methylene]- into a 50 mL round bottom flask. 1H-1,2,3-triazolo[4,5-b]pyridinium-3oxide-hexafluorophosphate (777'.0 mg, 2.04 mmol, 1.20 eq.), DMF (10 ml), N,N- diisopropylethylamine (550.2 mg, 4.26 mmol, 2.50 eq.) and (3S)-3-[[(tert-butyldimethylsilyl)oxy]methyl]morpholinecyclohexane (430.0 mg, 1.86 mmol, 1. 09 eq.). The resulting solution was stirred for 2 hours at room temperature. The reaction was then quenched by adding water. The resulting solution was extracted with 3x20 ml ethyl acetate, dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (0-50%) as eluent. (8)-3-(3-(3-(((tert-butyldimethylsilyl)oxy)methyl)morpholine-4-carbonyl)pyridin-2yl)propanenitrile was obtained. LCMS (ES, m/z): [M+H]+: 390.2.

Stage 5.

(S)-3-(3-(3-(((tertbutyldimethylsilyl)oxy)methyl)morpholine-4-carbonyl)pyridin-2-yl)propanenitrile (0.63 g, 1. 62 mmol, 1.00 eq.), THF (10 mL, 123.43 mmol, 76.3 eq.) and tetrabutylammonium fluoride (1.0 M) (2.43 mL, 2.43 mmol, 1.50 eq. .). The resulting solution was stirred for 2 hours at room temperature. The resulting mixture was concentrated and purified by silica gel column chromatography using dichloromethane/methanol (94.6:5.4) as eluent. (R)-3-(3-(3(hydroxymethyl)morpholine-4-carbonyl)pyridin-2-yl)propanenitrile was obtained. LCMS (ES, m/z): [M+H]+: 276.1.

Stage 6.

(R)-3-(3-(3-(hydroxymethyl)morpholine-4-carbonyl)pyridin-2-yl)propanenitrile (0.24 g) was placed in a 40 ml vial, which was purged and maintained under an inert atmosphere of nitrogen. , 0.87 mmol, 1.00 eq.), THF (10 ml), PPh ₃ (274.4 mg, 1.05 mmol, 1.20 eq.) and 2,6-dihydroxybenzaldehyde (156.5 mg, 1.13 mmol, 1.30 eq.). Di-tert-butyl azodicarboxylate (240.9 mg, 1.05 mmol, 1.20 eq) was then added dropwise at 0°C. After 20 minutes, the resulting solution was left with stirring at 40°C overnight. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (0-80%) as eluent. The crude product was purified by preparative-HPLC under the following conditions: column, Kinetex EVO C18, 21.2x150 mm, 5 µm; mobile phase; water (0.1% formic acid) and CH3CN (35% to 75% for 14 minutes). Detector; 220nm. Flow rate 20 ml/min. 3-{3-[(3S)-3-[(2formyl-3-hydroxyphenoxy)methyl]morpholine-4-carbonyl]pyridin-2-yl}propanenitrile was obtained. LCMS: (ES, m/z): [M+H]+: 396.2. 1H NMR: (300 MHz, DMSO-d ₆ , ppm): δ 11.72 (s, 1H), 10.17 (s, 1H), 8.62 (dd, J= 4.9, 1 .7 Hz, 1H), 7.78-7.36 (m, 3H), 6.76 (d, J= 8.3 Hz, 1H), 6.55 (d, J= 8.2 Hz, 1H ), 4.94-4.89 (m, 1H), 4.49-4.27 (m, 2H), 4.10-3.45 (m, 5H), 3.16-2.93 (m , 5H).

Example 25. 2-Hydroxy-6-{[(3 S)-4-[2-(2-methoxyethyl)benzoyl]morpholin-3-yl]methoxy}benzaldehyde, compound 29.

Compound 29 was synthesized according to Scheme 25.

Scheme 25

Stage 1.

Place 2-(2-methoxyethyl)benzoic acid (500.00 mg, 2.775 mmol, 1.00 eq), DCM (20.00 mL), (3R)-morpholin-3-ylmethanol ( 325.04 mg, 2.775 mmol, 1.00 eq.), 1 - [bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide-hexafluorophosphate ( 1582.51 mg, 4.162 mmol, 1.50 eq.) and N,N-diisopropylethylamine (1075.81 mg, 8.324 mmol, 3.00 eq.). The resulting solution was stirred for 3 hours at 25°C. The resulting mixture was concentrated. The residue was purified on a silica gel column using ethyl acetate/petroleum ether (1:1) as eluent. The collected fractions were combined and concentrated. [(3R)-4-[2-(2methoxyethyl)benzoyl]morpholin-3-yl]methanol was obtained. LCMS (ES) [M+1]+ m/z 280.2.

Stage 2.

[(3R)-4-[2-(2-methoxyethyl)benzoyl]morpholin-3yl]methanol (200.00 mg, 0.716 mmol, 1.00 eq.), tetrahydrofuran (10 mL) was placed in a 50 mL round bottom flask. , 2,6-dihydroxybenzaldehyde (98.89 mg, 0.716 mmol, 1.00 eq), triphenylphosphine (225.36 mg, 0.859 mmol, 1.20 eq) and diisopropyl azodicarboxylate (173.73 mg, 0.859 mmol, 1 ,20 eq.). The resulting solution was stirred for 16 hours at 25°C. The resulting mixture was concentrated. The crude reaction mixture was filtered and subjected to reverse phase preparative HPLC (Prep-C18, 20-45M, 120 g, Tianjin Bonna-Agela Technologies; gradient elution from 25% MeCN in water to 35% MeCN in water over a 10-minute period, where both solvents contained 0.1% formic acid) to obtain 2-hydroxy-6-{[(3S)4-[2-(2-methoxyethyl)benzoyl]morpholin-3-yl]methoxy}benzaldehyde. LCMS (ES) [M+1]+ m/z 400.2. 1H NMR (300 MHz, DMSO-Y6) δ 11.76 (broad s, 1H), 10.31 (s, 1H), 7.56 (t, J= 8.4 Hz, 1H), 7.46- 6.90 (m, 4H), 6.8-6.455 (m, 2H), 4.98-4.87 (m, 1H), 4.44-4.02 (m, 3H), 4.00- 3.27 (m, 8H), 3.15-2.55 (m, 4H).

Example 26. 2-Hydroxy-6-{[(3S)-4-[2-(2-methoxyethyl)benzyl]morpholin-3-yl]methoxy}benzaldehyde, compound 30.

Compound 30 was synthesized according to Scheme 26.

- 75 045055

Scheme 26

26b 26c

D

Stage 1.

Place 2-(2-cyanoethyl)pyridine-3-carboxylic acid (0.30 g, 1.70 mmol, 1.00 eq.), 1-[bis-(dimethylamino)methylene]-1H into a 50 mL round bottom flask -1,2,3-triazolo[4,5-b]pyridinium-3oxide-hexafluorophosphate (777.0 mg, 2.04 mmol, 1.20 eq.), DMF (10.0 ml), N,N- diisopropylethylamine (550.2 mg, 4.26 mmol, 2.50 eq.) and (2S)-2-[[(tert-butyldimethylsilyl)oxy]methyl]pyrrolidine (403.50 mg, 1.87 mmol, 1. 10 eq.). The resulting solution was stirred for 2 hours at room temperature. The reaction was then quenched by adding water (20 ml), extracted with 3x20 ml ethyl acetate, dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (0-60%) as eluent. (S)3-(3-(2-(((tert-butyldimethylsilyl)oxy)methyl)pyrrolidine-1-carbonyl)pyridin-2-yl)propanenitrile was obtained. LCMS (ES, m/z): [M+H]+: 374.2.

Stage 2.

(S)-3-(3-(2-(((tertbutyldimethylsilyl)oxy)methyl)pyrrolidin-1-carbonyl)pyridin-2-yl)propanenitrile (0.4 g, 1.07) was placed in a 50 ml round bottom flask. mmol, 1.0 equiv.), THF (10 ml) and tetrabutylammonium fluoride (1.2 ml, 1.20 mmol, 1.1 equiv.). The reaction solution was stirred for 2 hours at room temperature. The resulting mixture was concentrated and purified by silica gel column chromatography using MeOH/DCM (6:94) as eluent. (S)-3-(3-(2-(hydroxymethyl)pyrrolidin-1-carbonyl)pyridin-2-yl)propanenitrile was obtained. LCMS (ES, m/z): [M+H]+: 260.1.

Stage 3.

(S)-3-(3-(2-(hydroxymethyl)pyrrolidin-1-carbonyl)pyridin-2-yl)propanenitrile (120.00 mg) was placed in a 40 ml vial, which was purged and maintained under an inert atmosphere of nitrogen. , 0.46 mmol, 1.00 eq.), PPh ₃ (145.6 mg, 0.56 mmol, 1.20 eq.), THF (10 ml) and 1-(2,6dihydroxyphenyl)ethanone (91, 5 mg, 0.60 mmol, 1.30 eq.). Di-tert-butyl azodicarboxylate (127.9 mg, 0.56 mmol, 1.20 eq.) was then added dropwise at 0°C. After 20 minutes, the resulting solution was left with stirring at 40°C overnight. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (0-90%) as eluent. The crude product was purified by preparative-HPLC under the following conditions: column, Kinetex EVO C18, 21.2x150 mm, 5 µm; mobile phase water (0.1% formic acid) and CH3CN (40% to 70% for 14 minutes); detector, 220 nm. Flow rate 20 ml/min. 2-hydroxy-6-{[(3S)-4-[2-(2-methoxyethyl)benzoyl]morpholin-3-yl]methoxy}benzaldehyde was obtained. LCMS: (ES, m/z): [M+H]+: 380.2. ^1H NMR (300 MHz, DMSO^6, ppm): δ 11.68 (s, 1H), 10.34 (s, 1H), 8.61 (dd, J = 4.8, 1, 7 Hz, 1H), 7.72 (dd, J= 7.7, 1.8 Hz, 1H), 7.54 (t, J= 8.4 Hz, 1H), 7.38 (dd, J= 7.7, 4.8 Hz, 1H), 6.72 (d, J= 8.4 Hz, 1H), 6.55 (d, J= 8.4 Hz, 1H), 4.57-4, 53 (m, 1H), 4.3-4.29 (m, 2H), 4.10-3.59 (m, 1H), 3.32-3.14 (m, 2H), 3.06- 3.01 (m, 2H), 2.9-2.81 (m, 2H), 2.15-1.80 (m, 4H).

Example 27. 3-{2-[(3S)-3-[(2-formyl-3-hydroxyphenoxy)methyl]morpholine-4-carbonyl]phenyl}propanenitrile, compound 31.

Compound 31 was synthesized according to scheme 27.

- 76 045055

Scheme 27

Stage 1.

A 100 mL three-neck round bottom flask, purged and maintained under an inert nitrogen atmosphere, was charged with methyl 2-bromobenzoate (3.00 g, 13.951 mmol, 1.00 eq.), acrylonitrile (1.48 g, 27.901 mmol, 2.00 eq.), N,N-diisopropylethylamine (5.41 g, 41.859 mmol, 3.00 eq.), dioxane (50.00 ml) and Pd(P(t-Bu) ₃ ) ₂ (0. 71 g, 1.389 mmol, 0.10 eq.). The resulting solution was stirred for 16 hours at 100°C in an oil bath. The resulting mixture was concentrated. The residue was purified on a silica gel column using ethyl acetate/petroleum ether (10% ethyl acetate) as eluent. Methyl 2-[(1E)-2-cyanoeth-1-en-1-yl]benzoate was obtained. LCMS (ES) [M+1]+ m/z 188.0.

Stage 2.

A 100 mL round bottom flask was filled with methyl 2-[(1E)-2-cyanoeth-1-en-1-yl]benzoate (1.90 g, 10.150 mmol, 1.00 eq.), MeOH (40.00 ml, 987.956 mmol, 97.34 eq.) and Pd/C (0.80 g, 7.517 mmol, 0.74 eq.). The flask was evacuated and purged with nitrogen three times, followed by hydrogen. The mixture was stirred for 4 hours at room temperature under a hydrogen atmosphere (balloon). Solids were filtered off.

The filtrate was concentrated. Methyl 2-(2-cyanoethyl)benzoate was obtained. LCMS (ES) [M+1]+ m/z 190.1.

Stage 3.

Methyl 2-(2-cyanoethyl)benzoate (1.90 g, 10.042 mmol, 1.00 eq) and MeOH (50.00 mL) were placed in a 100 mL round bottom flask. A solution of LiOH (0.72 g, 30.065 mmol, 2.99 eq) in H2O (10 mL) was then added at 0°C for 5 minutes. The resulting solution was stirred for 16 hours at room temperature. The pH of the solution was adjusted to 5 using citric acid (3 mol/L). The resulting solution was extracted with 3x100 ml of DCM/MeOH=10:1 mixture. The mixture was dried over anhydrous sodium sulfate and concentrated in vacuo. 2-(2-cyanoethyl)benzoic acid was obtained. LCMS (ES) [M-1] - m/z 174.3.

Stage 4.

2-(2-cyanoethyl)benzoic acid (0.50 g, 2.854 mmol, 1.00 eq.), (3R)-morpholin-3-ylmethanol hydrochloride (0.66 g, 4.281 mmol) was placed in a 20 ml sealed tube. , 1.50 eq.), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide-hexafluorophosphate (1.63 g, 4.281 mmol, 1.50 eq.), X,X-diisopropylethylamine (1.11 g, 8.588 mmol, 3.01 eq.) and DMF (10.00 ml). The resulting solution was stirred for 2 hours at room temperature. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography using petroleum ether:THF (45% THF) as eluent. 3-[2-[(3R)-3(hydroxymethyl)morpholine-4-carbonyl]phenyl]propanenitrile was obtained. LCMS (ES) [M+1]+ m/z 275.1.

Stage 5.

3-[2-[(3R)-3-(hydroxymethyl)morpholine-4carbonyl]phenyl]propanenitrile (0.40 g, 1.458 mmol, 1 .00 eq.), 2,6-dihydroxybenzaldehyde (0.40 g, 2.916 mmol, 2 eq.), PPh ₃ (0.76 g, 2.916 mmol, 2 eq.) and THF (30.00 ml). The resulting solution was stirred for 15 minutes at 0°C. Diisopropyl azodicarboxylate (0.59 g, 2.918 mmol, 2.00 eq) was then added at 0°C for 3 minutes. The resulting solution was stirred for 2 hours at room temperature. The resulting mixture was concentrated. The residue was purified by column

- 77 045055 silica gel chromatography using petroleum ether/THF (22% THF) as eluent. The collected product was purified by flash preparative-HPLC (Prep-C18, 20-45M, 120 g, Tianjin Bonna-Agela Technologies; gradient elution from 45% MeCN in water to 65% MeCN in water over a 10-minute period, at In this case, both solvents contained 0.1% formic acid). 3-{2[(3S)-3-[(2-formyl-3-hydroxyphenoxy)methyl]morpholine-4-carbonyl]phenyl}propanenitrile was obtained. LCMS (ES) [M+1]+ m/z 395.2. ^1H NMR (300 MHz, DMSO- _d6 , ppm) δ 11.73 (s, 1H), 10.28 (s, 1H), 7.63-7.03 (m, 5H), 6 .77 (d, J= 8.3 Hz, 1H), 6.56 (d, J= 8.5 Hz, 1H), 5.06-4.22 (m, 3H), 4.12-3, 29 (m, 5H), 3.15-2.66 (m, 5H).

Example 28. 3-{3-[(3S)-3-[(2-Formyl-3-hydroxyphenoxy)methyl]morpholine-4-carbonyl]pyrazin-2yl}propanenitrile, compound 32.

Compound 32 was synthesized according to Scheme 28.

Scheme 28

Stage 1.

A 100 mL three-neck round bottom flask, purged and maintained under an inert nitrogen atmosphere, was charged with methyl 3-bromopyrazine-2-carboxylate (6.00 g, 27.647 mmol, 1.00 eq.), acrylonitrile (4.40 g 82.941 mmol, 3 eq.), X,X-diisopropylethylamine (10.72 g, 82.941 mmol, 3 eq.), dioxane (60.00 ml) and Pd(P(t-Bu) ₃ ) ₂ (1.41 g, 2.765 mmol, 0.1 eq.). The resulting solution was stirred for 16 hours at 100°C in an oil bath. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography using (10-20% ethyl acetate) as eluent. Methyl 3-[(1E)-2-cyanoeth-1-en-1-yl]pyrazin-2-carboxylate was obtained. LCMS (ES) [M+1]+ m/z 190.2.

Stage 2.

Methyl 3-[(1E)-2-cyanoeth-1-en-1-yl]pyrazin-2carboxylate (1.60 g, 8.458 mmol, 1.00 eq.), MeOH (20 .00 ml) and Pd/C (0.60 g, 5.638 mmol, 0.67 eq.). The flask was evacuated and purged with nitrogen three times, followed by hydrogen. The resulting solution was stirred for 4 hours at room temperature in a hydrogen atmosphere (balloon). Solids were filtered off. The filtrate was concentrated. Methyl 3-(2-cyanoethyl)pyrazine-2carboxylate was obtained. LCMS (ES) [M+1]+ m/z 192.2.

Stage 3.

Methyl 3-(2-cyanoethyl)pyrazine-2-carboxylate (0.70 g, 3.661 mmol, 1.00 eq) and MeOH (50 mL) were placed in a 100 mL round bottom flask. A solution of LiOH-H ₂ O (0.31 g, 7.387 mmol, 2.02 eq) in 10 ml H2O was then added dropwise with stirring at 0°C for 5 minutes. The resulting solution was stirred for 2 hours at room temperature. The pH of the solution was adjusted to 5 using citric acid (2 mol/L). The resulting solution was extracted with 20x100 ml of DCM/MeOH=10:1 and the organic layer was concentrated. 3-(2-cyanoethyl)pyrazine-2carboxylic acid was obtained. LCMS (ES) [M-1] ^- m/z 176.1.

Stage 4.

3-(2-cyanoethyl)pyrazin-2-carboxylic acid (0.43 g, 2.427 mmol, 1.00 equiv.), (3R)-morpholin-3-ylmethanol hydrochloride (0.34 g) was placed in a 20 ml vial , 2.913 mmol, 1.2 eq.), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide-hexafluorophosphate (1.11 g, 2.913 mmol, 1.20 eq.), X,X-diisopropylethylamine (0.94 g, 7.281 mmol, 3.00 eq.) and DMF (10.00 ml). The resulting solution was stirred for 2 hours at room temperature. The resulting mixture

- 78 045055 concentrated. The residue was purified by silica gel column chromatography using petroleum ether/THF (50% THF) as eluent. 3-[3-[(3R)-3(hydroxymethyl)morpholine-4-carbonyl]pyrazin-2-yl]propanenitrile was obtained. LCMS (ES) [M+1]+ m/z 277.1.

Stage 5.

3-[3-[(3R)-3-(hydroxymethyl)morpholine-4carbonyl]pyrazin-2-yl]propanenitrile (200.00 mg , 0.724 mmol, 1.00 eq.), 2,6-dihydroxybenzaldehyde (199.96 mg, 1.448 mmol, 2.00 eq.), PPh ₃ (379.72 mg, 1.448 mmol, 2.00 eq.) and THF (20.00 ml). The resulting solution was stirred for 15 minutes at 0°C. Diisopropyl azodicarboxylate (292.74 mg, 1.448 mmol, 2.00 eq.) was then added dropwise with stirring at 0° C. over 5 minutes. The resulting solution was stirred for 2 hours at room temperature. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography using petroleum ether/THF (35% THF) as eluent. The collected product was further purified by flash preparative-HPLC (Prep-C18, 20-45M, 120 g, Tianjin Bonna-Agela Technologies; gradient elution from 35% MeCN in water to 60% MeCN in water over a 10-minute period, where both solvents contained 0.1% formic acid). Received

3-[3-[(3S)-3-(2-formyl-3hydroxyphenoxymethyl)morpholine-4-carbonyl]pyrazin-2-yl]propanenitrile. LCMS (ES) [M+1]+ m/z 397.2. 1H NMR (300 MHz, DMSOL, ppm) δ 11.75 (s, 1H), 10.32 (s, 1H), 8.72 (dd, J= 8.2, 2.5 Hz, 1H ), 8.55 (dd, J= 13.7, 2.5 Hz, 1H), 7.62-7.51 (m, 1H), 6.80-6.48 (m, 2H), 5, 03-4.94 (m, 1H), 4.53-4.31 (m, 2H), 4.12-3.38 (m, 5H), 3.21-3.04 (m, 3H), 2.99-2.90 (m, 2H).

Example

29.

2-Hydroxy-6-{[(38)-4-[3-(hydroxymethyl)pyrazine-2-carbonyl]morpholin-3 yl]methoxy}benzaldehyde, compound 33.

Compound 33 was synthesized according to Scheme 29.

Scheme 29

Stage 1.

In a round bottom flask with a volume of

Methyl 3-bromopyrazine-2-carboxylate (5.00 g, 23.039 mmol, 1.00 eq.), H2O (100.00 mL) was added to 250 mL, which was purged and maintained under an inert nitrogen atmosphere. NaBH ₄ (4.36 g, 115.243 mmol, 5.00 eq.) was then added in several portions at 0°C. The resulting solution was stirred overnight at room temperature. The reaction was quenched by adding 50 ml EtOH and diluted with 150 ml K2CO3 (aq). The mixture was stirred for 0.5 hour at room temperature. The resulting solution was extracted with 3x150 ml ethyl acetate and extracted with 3x150 ml dichloromethane; the organic layer was dried over anhydrous sodium sulfate and concentrated. (3-bromopyrazin-2-yl)methanol was obtained. LCMS (ES) [M+1]+ m/z 189.1.

Stage 2.

Place (3-bromopyrazin-2-yl)methanol (3.00 g, 15.872 mmol, 1.00 eq), DCM (60.00 mL), imidazole (2.16 g, 31.729 mmol) into a 100 mL round bottom flask , 2.00 eq.) and tert-butyldimethylsilyl chloride (2.87 g, 19.042 mmol, 1.20 eq.). The resulting solution was stirred for 3 hours at room temperature and diluted with 50 ml of H2O. The resulting solution was extracted with 3x150 ml dichloromethane; the organic layer was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (20% ethyl acetate) as eluent. 2-bromo-3-[[(tert-butyldimethylsilyl)oxy]methyl]pyrazine was obtained.

LCMS (ES) [M+1]+ m/z 303.1.

Stage 3.

2-Bromo-3-[[(tertbutyldimethylsilyl)oxy]methyl]pyrazine (6.00 g, 19.784 mmol, 1.00 eq.), Pd(dppf)Cl ₂ (1.45 g, 1.978 mmol, 0.10 eq.), triethylamine (6.01 g, 59.352 mmol, 3.00 eq.), MeOH (100.00 ml) and CO (gas). The resulting solution was stirred overnight at 90°C. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography using petroleum ether/THF (70% THF) as eluent. Methyl 3-[[(tert-butyldimethylsilyl)oxy]methyl]pyrazine-2carboxylate was obtained. LCMS (ES) [M+1] ⁺ m/z 283.2.

Stage 4.

Methyl 3-[[(tert-butyldimethylsilyl)oxy]methyl]pyrazine-2-carboxylate (3.10 g, 10.977 mmol, 1.00 eq.), methanol (50.00 mL) was placed in a 250 mL round bottom flask. A solution of LiOH-H ₂ O (0.92 g, 21.924 mmol, 2.00 eq) in H2O (10 ml) was then added dropwise with stirring at 0° C. over 5 minutes. The resulting solution was stirred for 5 hours at room temperature. The pH of the solution was adjusted to 5 using citric acid (2 mol/L). The resulting solution was extracted with 5x150 ml of dichloromethane; the organic layer was dried over anhydrous sodium sulfate and concentrated. The result was 3-[[(tert-butyldimethylsilyl)oxy]methyl]pyrazine-2-carboxylic acid. LCMS (ES) [M+1] ⁺ m/z 269.2.

Stage 5.

3-[[(tert-butyldimethylsilyl)oxy]methyl]pyrazin-2-carboxylic acid (1.00 g, 3.726 mmol, 1.00 eq.), (3R)-morpholin-3-ylmethanol was placed in a 20 mL vial. hydrochloride (0.68 g, 4.471 mmol, 1.20 eq.), dimethylformamide (10.00 ml), 1-[bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5b] pyridinium 3-oxide hexafluorophosphate (1.70 g, 4.471 mmol, 1.20 eq.) and K, K-diisopropylethylamine (1.95 ml, 15.064 mmol, 3.00 eq.). The resulting solution was stirred for 3 hours at room temperature. The resulting solution was diluted with 50 ml H ₂ O. The resulting solution was extracted with 4x60 ml dichloromethane and the organic layer was washed with 2x100 ml brine. The mixture was dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography using petroleum ether/THF (60% THF) as eluent. [(3R)-4-(3-[[(tertbutyldimethylsilyl)oxy]methyl]pyrazin-2-carbonyl)morpholin-3-yl]methanol was obtained. LCMS (ES) [M+1] ⁺ m/z 368.2.

Stage 6.

[(3R)-4-(3-[[(tertbutyldimethylsilyl)oxy]methyl]pyrazin-2-carbonyl)morpholin-3-yl] was placed into a 100 ml three-neck round-bottom flask, which was purged and maintained inside an inert atmosphere of nitrogen. methanol (0.97 g, 2.639 mmol, 1.00 eq.), 2,6-dihydroxybenzaldehyde (0.73 g, 5.279 mmol, 2.00 eq.), PPh ₃ (1.38 g, 5.261 mmol, 1 .99 eq.) and THF (60 ml). The resulting solution was stirred for 15 minutes at 0°C. Diisopropyl azodicarboxylate (1.07 g, 5.279 mmol, 2.00 eq.) was then added dropwise with stirring at 0° C. over 2 minutes. The resulting solution was stirred for 2 hours at room temperature. The resulting mixture was concentrated. The residue was purified on a silica gel column using petroleum ether/THF (50% THF) as eluent. 2-[[(3S)-4-(3-[[(tert-butyldimethylsilyl oxy]methyl]pyrazin-2-carbonyl)morpholin-3-yl]methoxy]-6-hydroxybenzaldehyde was obtained. LCMS (ES) [M+ 1] ⁺ m/z 488.2.

Stage 7.

2-[[(3S)-4-(3-[[(tertbutyldimethylsilyl)oxy]methyl]pyrazin-2-carbonyl)morpholin-3-yl]methoxy]-6-hydroxybenzaldehyde (0. 8 g, 1.641 mmol, 1.00 eq.), THF (20.00 mL) and tetrabutylammonium fluoride (2.5 mL, 1.5 eq., 2M). The resulting solution was stirred for 2 hours at room temperature. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography with (60% THF) as eluent. The collected product was further purified by flash preparative-HPLC (Prep-C18, 20-45M, 120 g, Tianjin Bonna-Agela Technologies; gradient elution from 30% MeCN in water to 50% MeCN in water over a 10-minute period, where both solvents contained 0.1% formic acid). 2-hydroxy-6-{[(3S)-4-[3-(hydroxymethyl)pyrazine-2-carbonyl]morpholin3-yl]methoxy}benzaldehyde was obtained. LCMS (ES) [M+1] ⁺ m/z 374.1. 1H NMR (300 MHz, DMSO-d ₆ , ppm) δ 11.79 (s, 1H), 10.29 (d, J= 3.6 Hz, 1H), 8.64 (t, J= 2.4 Hz, 1H), 8.54 (dd, J= 12.2, 2.6 Hz, 1H), 7.62-7.49 (m, 1H), 6.776.48 (m, 2H), 5.72-5.51 (m, 1H), 4.95-4.30 (m, 5H), 4.12-3.39 (m, 5H), 3.24-2.97 (m, 1H ).

Example 30. 2-{[(2S)-1-[2-(1,2-Dihydroxyethyl)benzoyl]piperidin-2-yl]methoxy}-6-hydroxybenzaldehyde, compound 34.

Compound 34 was synthesized according to scheme 30.

- 80 045055

Scheme 30

Stage 1.

O-bromobenzoic acid (5.0 g, 24.87 mmol, 1.0 equiv.), (2S)-2-[[(tert-butyldimethylsilyl)oxy]methyl]piperidine (6 .90 g, 30.07 mmol, 1.2 eq.), DCM (50.0 mL) and N,N-diisopropylethylamine (6.50 g, 50.29 mmol, 2.0 eq.). After this, 1-[bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide-hexafluorophosphate (11.40 g, 29.98 mmol, 1. 2 equiv.) at 0°C. The reaction solution was stirred for 2 hours at room temperature. The reaction was then quenched by adding water (30 ml) and extracted with 2x50 ml dichloromethane. The residue was purified on a silica gel column using ethyl acetate/petroleum ether (10%) as eluent. (8)-(2-bromophenyl)(2-(((tert-butyldimethylsilyl)oxy)methyl)piperidin-1 yl)methanone was obtained. LCMS (ES) [M+1]+ m/z: 412.

Stage 2.

(8)-(2-bromophenyl)(2-(((tertbutyldimethylsilyl)oxy)methyl)piperidin-1-yl)methanone (5.0 g, 12.12 mmol, 1.0 equiv) was placed in a 250 mL round bottom flask .), THF (50 ml) and 1 M tetrabutylammonium fluoride in THF (12.1 ml, 12.12 mmol, 1.0 eq.). The mixture was stirred for 2 hours at room temperature. The mixture was concentrated to remove the solvent and the residue was purified on a silica gel column with ethyl acetate (100%) as eluent. (S)-(2bromophenyl)(2-(hydroxymethyl)piperidin-1-yl)methanone was obtained. LCMS (ES) [M+1]+ m/z: 298.

Stage 3.

(8)-(2-bromophenyl)(2-(hydroxymethyl)piperidin-1yl)methanone (2.0 g, 6.71 mmol, 1.0 eq), 2,6-dihydroxybenzaldehyde (1.12 g, 8.11 mmol, 1.2 eq), THF (80 ml) and PPh ₃ (2.10 g, 8.01 mmol, 1 ,2 eq.). Diisopropyl azodicarboxylate (1.63 g, 8.05 mmol, 1.2 eq.) was then added at 0°C. The reaction solution was stirred overnight at room temperature. The solution was concentrated to remove the solvent and the residue was purified on a silica gel column using ethyl acetate/petroleum ether (80%) as eluent. (S)-2-((1-(2bromobenzoyl)piperidin-2-yl)methoxy)-6-hydroxybenzaldehyde was obtained. LCMS (ES) [M+1]+ m/z: 418.

Stage 4.

(S)-2-((1-(2-bromobenzoyl)piperidin-2-yl)methoxy)-6hydroxybenzaldehyde (1.57 g, 3 .75 mmol, 1.0 eq.), dioxane (20 ml), tributyl(ethenyl)stannane (2.40 g, 7.54 mmol, 2.0 eq.) and Pd(dppf)Cl ₂ (307 mg, 0.37 mmol, 0.10 eq.). The mixture was stirred overnight at 90°C. The mixture was concentrated to remove the solvent and the residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1/1) as eluent. (8)-2-hydroxy-6-((1-(2-vinylbenzoyl)piperidin-2-yl)methoxy)benzaldehyde was obtained. LCMS (ES) [M+1]+ m/z: 366.

Stage 5.

(8)-2-hydroxy-6-((1-(2-vinylbenzoyl)piperidin-2yl)methoxy)benzaldehyde (604 mg, 1.65 mmol, 1.0 eq.), t- BuOH (4.0 ml), H ₂ O (4.0 ml) and ad-mix-alpha (2.60 g, 4.96 mmol, 3.0 eq.). The mixture was stirred for 3 hours at room temperature. The crude product was purified by preparative-HPLC under the following conditions (SHIMADZU (HPLC-01)): column, Kinetex EVO C18 column, 21.2x150, 5 µm, mobile phase, water (0.1% formic acid) and CH ₃ CN (45% to 65% phase B for 9 minutes), detector, UV 254 nm. 2{[(2S)-1-[2-(1,2-dihydroxyethyl)benzoyl]piperidin-2-yl]methoxy}-6-hydroxybenzaldehyde was obtained. HCMS:

(ES, m/z): [M+H]+: 400.2. ^1H NMR: (300 MHz, DMSO-66, ppm): δ 11.73 (s, 1H), 10.26 (s, 1H), 7.59-7.20 (m,

5H), 6.88-6.53 (m, 2H), 5.26-4.27 (m, 6H), 3.55-3.05 (m, 4H), 1.94-1.43 ( m, 6H).

Example 31 {2-[(1K)-1,2-dihydroxyethyl]benzoyl}thiomorpholin-3-yl]methoxy}-6hydroxybenzaldehyde.

Compound 35, diastereomer 1 and compound 35, diastereomer 2 were synthesized according to Scheme 31.

Scheme 31

Connection Bid

Diastereomer! and Diastereomer 2

Connection 35

Dgastereomer! iDgastereomer2

Stage 1.

Place 2-(2,2-dimethyl-1,3-dioxolan-4-yl)benzoic acid (900 mg, 4.05 mmol, 1.00 eq.), DMF (10.0 mL) into a 100 mL round bottom flask ), (3R)-thiomorpholin-3-ylmethanol (593 mg, 4.45 mmol, 1.10 eq.) and N,N-diisopropylethylamine (1.05 g, 8.09 mmol, 2.00 eq.). After this, 1-[bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide-hexafluorophosphate (2.31 g, 6.07 mmol, 1. 50 equiv.) at 0°C. The resulting solution was stirred for 2 hours at room temperature. The resulting solution was diluted with 50 ml H ₂ O and extracted with 3x30 ml ethyl acetate, the organic layers were combined, dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1/10) as eluent. [(3R)-4-[2-(2,2-dimethyl-1,3-dioxolan-4-yl)benzoyl]thiomorpholin-3yl]methanol was obtained. [M+1]+ m/z: 338.1.

Stage 2.

[(3R)-4-[2-(2,2-Dimethyl-1,3-dioxolan-4yl)benzoyl]thiomorpholin-3-yl]methanol (620 mg, 1.83 mmol, 1.00 eq), dichloroethane (8.0 ml) and N,Ndiisopropylethylamine (1.42 g, 11.02 mmol, 6.00 eq). Methylsulfonyl (420 mg, 3.67 mmol, 2.00 eq) was then added dropwise with stirring at 0°C. The resulting solution was stirred for 1 hour at room temperature. To this solution was added 2,6-dihydroxybenzaldehyde (253 mg, 1.83 mmol, 1.00 eq.). The resulting solution was stirred for 6 hours at 80°C. The reaction mixture was cooled and concentrated. The resulting solution was diluted with 5 ml of acetonitrile. The crude product was purified by flash preparative-HPLC under the following conditions (IntelFlash-1): column, silica gel C18; mobile phase, H ₂ O (0.1% HCOOH)/acetonitrile = 1/1, increasing to H ₂ O (0.1% HCOOH)/acetonitrile = 1/2 over 10 minutes; detector, UV 254 nm. 2-[[(3R)-4-[2-(2,2dimethyl-1,3-dioxolan-4-yl)benzoyl]thiomorpholin-3-yl]methoxy]-6-hydroxybenzaldehyde was obtained. [M+1]+ m/z: 458.2.

Stage 3.

2-Hydroxy-6-[[(3R)-4-[2-(2-methyl-1,3-dioxolan-4-yl)benzoyl]thiomorpholin-3-yl]methoxy]benzaldehyde was purified using a chiral preparative method. HPLC under the following conditions: mobile phase

- 82 045055

A: n-hexane; mobile phase B: EtOH; flow rate: 20 ml/min; column: DAICEL CHIRALPAK ID, 250*20 mm, 5 µm; gradient: 5% B for 20 minutes; detector, UV 254 nm. The collected products were subjected to chiral HPLC analysis (instrument name: Shimadzu LC-20AD; mobile phase A: n-hexane; mobile phase B: ethanol; column: CHIRALPAK IC-3, 50*4.6 mm, 3 μm IC30CC-SC002) Obtained compound 31d, diastereomer 2 (analytical HPLC retention time=2.188 min) and compound 31d, diastereomer 1 (analytical HPLC retention time=2.988 min).

Stage 4A. Compound 35, diastereomer 1.

Compound 31, diastereomer 1 (80 mg, 0.17 mmol, 1.00 eq.), acetonitrile (2.0 ml), Yb(OTf) ₃ H ₂ O (54 mg, 0 .08 mmol, 0.50 eq.). The resulting solution was stirred for 5 hours at room temperature. The resulting solution was diluted with 5 ml of acetonitrile and filtered. The crude product was purified by preparative-HPLC under the following conditions (2#SHIMADZU (HPLC-01)): column, Welch Xtimate C18, 21.2x250 mm, 5 µm; mobile phase, water and acetonitrile (15% to 70% phase B for 20 minutes); detector, UV 254 nm. The product was analyzed using chiral chromatography with supercritical mobile phase (instrument name: Shimadzu LC-30AD SF; column: AS-3, 100*3 mm). Compound 35, diastereomer 1 was obtained. Chiral chromatography retention time with supercritical mobile phase = 2.75 min. LCMS [M+1]+ m/z: 418.2. ^1H NMR (300 MHz, DMSO-d^) δ 11.76 (broad s, 1H), 10.43-10.06 (m, 1H), 7.64-6.88 (m, 5H), 6 .87-6.45 (m, 2H), 5.57-5.15 (m, 2H), 4.93-4.37 (m, 4H), 3.68-3.37 (m, 3H) , 3.24-2.83 (m, 2H), 2.83-2.59 (m, 2H), 2.48-2.22 (m, 1H).

Stage 4B. Compound 35, diastereomer 2.

Compound 31d, diastereomer 2 (90 mg, 0.19 mmol, 1.00 eq), acetonitrile (2.0 ml) and Yb(OTf) ₃ -H ₂ O (61 mg, 0) were placed in a 50 ml round bottom flask. .09 mmol, 0.50 eq.). The resulting solution was stirred for 5 hours at room temperature. The resulting solution was diluted with 5 ml of acetonitrile and filtered. The crude product was purified by preparative-HPLC under the following conditions (2#SHIMADZU (HPLC-01)): column, Welch Xtimate C18, 21.2x250 mm, 5 µm; mobile phase, water and acetonitrile (15% to 70% phase B over 18 minutes); detector, UV 254 nm. The product was analyzed using chiral chromatography with supercritical mobile phase (instrument name: Shimadzu LC-30AD SF; column: AS-3, 100*3 mm). Compound 35, diastereomer 2 was obtained. Chiral chromatography retention time with supercritical mobile phase = 2.44 min. LCMS [M+1]+ m/z: 418.2. 1H NMR (300 MHz, DMSO- _d6 ) δ 11.76 (broad s, 1H), 10.4610.03 (m, 1H), 7.70-6.90 (m, 5H), 6.82- 6.48 (m, 2H), 5.52-5.15 (m, 2H), 4.93-4.02 (m, 4H), 3.63-3.36 (m, 3H), 3. 26-2.90 (m, 2H), 2.88-2.55 (m, 2H), 2.48-2.24 (m, 1H).

Example 32. 2-{[(2S)-1-[2-(1,2-Dihydroxyethyl)pyridin-3-carbonyl]piperidin-2-yl]methoxy}-6hydroxybenzaldehyde, compound 36.

Compound 36 was synthesized according to Scheme 32.

Scheme 32

D.H.

Stage 1.

Place 2-bromopyridine-3-carboxylic acid (4.0 g, 19.80 mmol, 1.0 eq.), (2S)-2-[[(tert-butyldimethylsilyl)oxy]methyl into a 100 mL three-neck round bottom flask. ]piperidine (5.50 g, 23.97 mmol, 1.2 eq.), DCM (50 ml) and N,N-diisopropylethylamine (5.13 g, 39.70 mmol, 2.0 eq.). 1-[Bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxidehexafluorophosphate (9.07 g, 23.85 mmol, 1.2 eq) was then added .) at 0°C. The reaction solution was stirred for 2 hours at room temperature. The reaction was then quenched by adding water (50 ml) and extracted with 2x50 ml dichloromethane. The combined organic phase was dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The residue was purified on a silica gel column with ethyl acetate/petroleum ether (1/4). (S)-(2-bromopyridin-3-yl)(2-(((tertbutyldimethylsilyl)oxy)methyl)piperidin-1-yl)methanone was obtained. LCMS (ES) [M+1]+ m/z: 413.

- 83 045055

Stage 2.

(S)-(2-bromopyridin-3-yl)(2-(((tertbutyldimethylsilyl)oxy)methyl)piperidin-1-yl)methanone (8.0 g, 19.35 mmol, 1.0 eq.), THF (80 ml) and tetrabutylammonium fluoride (1M in THF) (20 ml, 20.0 mmol, 1.0 eq.). The mixture was stirred for 2 hours at room temperature and concentrated to remove the solvent. The residue was purified by silica gel column chromatography using ethyl acetate (100%) as eluent. (S)-(2-bromopyridin-3-yl)(2-(hydroxymethyl)piperidin-1-yl)methanone was obtained. LCMS (ES) [M+1]+ m/z: 299.

Stage 3.

(S)-(2-bromopyridin-3-yl)(2-(hydroxymethyl)piperidin-1yl)methanone (2.0 g, 6 .69 mmol, 1.0 eq.), 2,6-dihydroxybenzaldehyde (1.10 g, 7.96 mmol, 1.2 eq.), PPh ₃ (2.10 g, 8.01 mmol, 1.2 eq.) and THF (80 ml). Diisopropyl azodicarboxylate (1.63 g, 8.06 mmol, 1.2 eq.) was then added at 0°C. After addition, the resulting solution was stirred overnight at room temperature. The mixture was concentrated to remove the solvent and the residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1/1) as eluent. 2-(S)-2-(( 1 -(2-bromonicotinoyl)piperidin-2-yl)methoxy)-6-hydroxybenzaldehyde was obtained. LCMS (ES) [M+1]+ m/z: 419.

Stage 4.

2-(S)-2-((1-(2-bromonicotinoyl)piperidin-2-yl)methoxy)-6hydroxybenzaldehyde (3.20 g) was placed in a 100 ml round bottom flask, which was purged and maintained under an inert atmosphere of nitrogen. , 7.63 mmol, 1.0 eq.), dioxane (30 ml), tributyl(ethenyl)stannane (4.85 g, 15.30 mmol, 2.0 eq.) and Pd(dppf)Cl ₂ -CH ₂ Cl ₂ (624 mg, 0.76 mmol, 0.10 eq.). The mixture was stirred overnight at 90°C. After cooling to room temperature, the reaction solution was concentrated to remove the solvent, and the residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1/1) as the eluent. (S)-2-hydroxy-6-((1-(2vinylnicotinoyl)piperidin-2-yl)methoxy)benzaldehyde was obtained. LCMS (ES) [M+1]+ m/z: 367.

Stage 5.

(S)-2-hydroxy-6-((1-(2vinylnicotinoyl)piperidin-2-yl)methoxy)benzaldehyde (500 mg, 1.37 mmol, 1.0 eq.) was placed in a 100 ml round bottom flask, t -BuOH (20.0 ml), H ₂ O (20.0 ml) and AD-mix-alpha (5.31 g, 6.82 mmol, 5.0 eq.). The mixture was stirred overnight at room temperature. The mixture was concentrated to remove the solvent and the crude product was purified by preparative-HPLC under the following conditions (SHIMADZU (HPLC-01): column, Kinetex EVO C18 column, 21.2x150, 5 µm, mobile phase, water (0.1% formic acid ) and CH3CN (45% to 65% phase B in 9 minutes), detector, UV 254 nm. 2-{[(2S)-1-[2-(1,2dihydroxyethyl)pyridine-3-carbonyl]piperidine was obtained -2-yl]methoxy}-6-hydroxybenzaldehyde LCMS: (ES, m/z): [M+H]+: 401.2 1H NMR (300 MHz, DMSO-d ₆ , ppm): δ 11.82-11.66 (m, 1H), 10.34-10.14 (m, 1H), 8.56 (d, 1H, J= 1.8 Hz), 7.68-7.31 (m, 3H), 6.78-6.54 (m, 2H), 5.22-5.19 (m, 2H), 4.71-4.29 (m, 4H), 3.68-3 .58 (m, 2H), 3.19-2.90 (m, 2H), 2.08-1.50 (m, 6H).

Example 33. 2-Hydroxy-6-{[(3R)-4-[2-(2-hydroxyethyl)pyridine-3-carbonyl]morpholin-3yl]methoxy}benzaldehyde, compound 37.

Compound 37 was synthesized according to Scheme 33.

Scheme 33

33a HE 0С & Stage 2 Stage 1. In a three-necked round bottom zz ^Chn OTBS _he %_. f Stage 3 ⁰ 0^ ^CH0 X χ:ι yu os a _he 33c 37 a 50 ml flask was placed 2-[2-[(tert-

- 84 045055 butyldimethylsilyl)oxy]ethyl]pyridine-3-carboxylic acid (1.50 g, 5.33 mmol, 1.0 eq.), (3S)morpholin-3-ylmethanol hydrochloride (980 mg, 6.38 mmol , 1.2 eq.), DCM (15 ml) and N,Ndiisopropylethylamine (2.07 g, 16.02 mmol, 3.0 eq.). 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide-hexafluorophosphate (2.40 g, 6.31 mmol, 1.2 eq.) at 0°C. After addition, the mixture was stirred for 2 hours at room temperature. The reaction was then quenched by adding water (20 ml) and extracted with 3x20 ml dichloromethane. The combined organic phase was dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (80%) as eluent. (S)-(2-(2((tert-butyldimethylsilyl)oxy)ethyl)pyridin-3-yl)(3-(hydroxymethyl)morpholino)methanone was obtained. LCMS (ES) [M+1]+ m/z: 381.

Stage 2.

(S)-(2-(2-((tert-butyldimethylsilyl)oxy)ethyl)pyridin-3yl)(3-(hydroxymethyl)morpholino )methanone (1.98 g, 5.20 mmol, 1.0 eq.), 2,6-dihydroxybenzaldehyde (863 mg, 6.25 mmol, 1.2 eq.), PPh ₃ (1.64 g, 6 .25 mmol, 1.2 eq.) and THF (80 ml). After cooling to 0° C., di-tert-butyl azodicarboxylate (1.44 g, 6.25 mmol, 1.2 eq.) was added in one portion. The mixture was stirred overnight at room temperature. The mixture was concentrated to remove the solvent and the residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (80%) as eluent. (R)-2-((4-(2-(2-((tertbutyldimethylsilyl)oxy)ethyl)nicotinoyl)morpholin-3-yl)methoxy)-6-hydroxybenzaldehyde was obtained. LCMS (ES) [M+1]+ m/z: 501.

Stage 3.

(R)-2-((4-(2-(2-((tert-butyldimethylsilyl)oxy)ethyl)nicotinoyl)morpholin-3-yl)methoxy)-6-hydroxybenzaldehyde (300 mg, 0.60 mmol, 1.0 eq.), CH3CN (5.0 ml) and HCOOH (1.0 ml). The reaction solution was stirred for 1 hour at 50°C and then concentrated to remove the solvent. The residue was purified by preparative HPLC under the following conditions: Kinetex EVO C18 column, 21.2x150, 5 µm, mobile phase, water (0.1% formic acid) and CH3CN (10% to 50% phase B over 15 minutes), detector, UV 254 nm. 2-hydroxy-6-{[(3R)-4-[2-(2-hydroxyethyl)pyridin-3-carbonyl]morpholin-3yl]methoxy}benzaldehyde was obtained. LCMS (ES, m/z): [M+H]+: 387.1. 1H NMR (300 MHz, DMSO- _d6 , ppm): δ 11.74 (broad s, 1H), 10.33-10.12 (m, 1H), 8.56 (dd, J=4 .8, 1.8 Hz, 1H), 7.71-7.29 (m, 3H), 6.78-6.52 (m, 2H), 5.034.89 (m, 1H), 4.65- 4.34 (m, 3H), 4.11-3.34 (m, 7H), 3.11-2.84 (m, 3H).

Example 34. 2-G hydroxy-6-{[(2R)-1 -[2-(2-hydroxyethyl)pyridin-3-carbonyl]piperidin-2yl]methoxy}bechaldehyde, compound 38.

Compound 38 was synthesized according to Scheme 34.

Scheme 34

34a 34b

34s star

Stage 1.

2-[2-[(tert-butyldimethylsilyl)oxy]ethyl]pyridine-3carboxylic acid (1.00 g, 3.55 mmol, 1 .00 eq.), DMF (25.0 ml), (2R)-piperidin-2-ylmethanol (491 mg, 4.26 mmol, 1.20 eq.) and N,N-diisopropylethylamine (551 mg, 4.0 eq.), 26 mmol, 1.20 eq.). After this, 1 -[bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3 oxide hexafluorophosphate (1.62 g, 4.26 mmol, 1 .2 eq.) at 0°C. The reaction solution was stirred overnight at room temperature. The reaction mixture was diluted with 30 ml H ₂ O and extracted with 3x100 ml ethyl acetate. The combined organic phase was washed with 1 x 50 ml brine and dried over anhydrous sodium sulfate. The mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1:2) as eluent. [(2R)-1-(2-[2-[(tertbutyldimethylsilyl)oxy]ethyl]pyridin-3-carbonyl)piperidin-2-yl]methanol was obtained. LCMS (ES) [M+1]+ m/z:

379.

Stage 2.

[(2R)-1-(2-[2-[(tert-butyldimethylsilyl)oxy]ethyl]]pyridine-3-carbonyl)piperidine was placed in a 100 mL three-neck round-bottom flask, which was purged and maintained under an inert nitrogen atmosphere. -2-yl]methanol (1.20 g, 3.17 mmol, 1.00 eq.), THF (50.0 mL), 2,6-dihydroxybenzaldehyde (525 mg, 3.80 mmol, 1.20 eq. .) and PPh ₃ (998 mg,

3.80 mmol, 1.20 eq.). A solution of diisopropyl azodicarboxylate (769 mg, 3.80 mmol, 1.20 eq) in THF (2.00 mL) was then added dropwise with stirring at 0°C. The reaction solution was stirred overnight at room temperature. After concentration under reduced pressure, the residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1:1) as eluent. 2-[[(2R)-1-(2-[2-[(tert-butyldimethylsilyl)oxy]ethyl]pyridine3-carbonyl)piperidin-2-yl]methoxy]-6-hydroxybenzaldehyde was obtained. LCMS (ES) [M+1]+ m/z: 499.

Stage 3.

2-[[(2R)-1-(2-[2-[(tert-butyldimethylsilyl)oxy]ethyl]pyridin-3-carbonyl)piperidin-2-yl]methoxy]-6- was placed in a 50 ml round bottom flask. hydroxybenzaldehyde (300 mg, 0.60 mmol, 1.00 eq), CH3CN (5.00 ml) and HCOOH (1.00 ml). The mixture was stirred for 3 hours at 50°C in an oil bath. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was purified by flash preparative-HPLC under the following conditions (IntelFlash-1): column: Ascentis Express C18, 50*3.0 mm, 2.7 µm, mobile phase A: water/0.05% formic acid, mobile phase B: CH3CN; flow rate: 1.5 ml/min, gradient: 5% to 100% B over 1.2 minutes, retention 0.6 minutes. 2-hydroxy-6-{[(2R)-1-[2-(2hydroxyethyl)pyridin-3-carbonyl]piperidin-2-yl]methoxy}benzaldehyde was obtained. LCMS (ES, m/z): [M+H]+: 385. Ή NMR (300 MHz, DMSO- _d6 , ppm): δ 11.73 (broad s, 1H), 10.22 ( s, 1H), 8.54 (s, 1H), 7.69-7.23 (m, 3H), 6.75 (d, J= 8.4 Hz, 2H), 5.21-5.20 (m, 1H), 4.65-4.27 (m, 3H), 3.78-3.65 (m, 2H)), 3.20-2.68 (m, 4H), 1.951.39 ( m, 6H).

Example 35. 2-Hydroxy-6-{[(3S)-4-[2-(2-hydroxy-2-methylpropyl)pyridin-3-carbonyl]morpholin-3yl]methoxy}bechaldehyde, compound 39.

Compound 39 was synthesized according to Scheme 35.

Scheme 35

Stage 1.

3-Bromo-2-methylpyridine (20.0 g, 116.26 mmol, 1.0 eq.) and THF (400 mL) were placed in a 1 L, three-neck round bottom flask, purged and maintained under an inert nitrogen atmosphere. Lithium diisopropylamide (2M in THF) (69.8 mL, 139.51 mmol, 1.2 eq) was then added at -78°C and stirred for 0.5 hour. Acetone (7.46 g, 128.45 mmol, 1.1 eq) was added to the mixture at the same temperature. The mixture was stirred for 1 hour at -78°C. Then the reaction was quenched

- 86 045055 by adding NH ₄ Cl (aq) (300 ml) and extracted with 3x500 ml ethyl acetate. The combined organic phase was dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (15%) as eluent. 1-(3-bromopyridin-2-yl)-2methylpropan-2-ol was obtained. LCMS (ES) [M+1]+ m/z: 230.

Stage 2.

1-(3-bromopyridin-2-yl)-2-methylpropan-2-ol (4.0 g, 17.38 mmol, 1.0 eq.), tert-butyldimethylsilyl chloride (3 .10 g, 20.86 mmol, 1.2 eq.), DMF (40 ml), imidazole (2.38 g, 34.76 mmol, 2.0 eq.) and 4-dimethylaminopyridine (212 mg, 1.0 eq.) 74 mmol, 0.10 eq.). The reaction solution was stirred for 24 hours at 60°C. After cooling to room temperature, the reaction was quenched by adding water (50 ml) and extracted with 3x50 ml ethyl acetate. The combined organic phase was washed with 2x50 ml brine and dried over anhydrous sodium sulfate. The mixture was filtered and the filtrate was concentrated under reduced pressure; the residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1/20) as eluent. 3-bromo-2-(2-((tert-butyldimethylsilyl)oxy)-2methylpropyl)pyridine was obtained. LCMS (ES) [M+1] ⁺ m/z: 344.

Stage 3.

3-Bromo-2-(2-((tertbutyldimethylsilyl)oxy)-2-methylpropyl)pyridine (4.18 g, 12.14 mmol, 1.0 eq.), MeOH (80 ml), triethylamine (2.45 g, 24.28 mmol, 2.0 eq.) and Pd(dppf)Cl ₂ CH ₂ Cl ₂ (495 mg, 0.61 mmol, 0.05 eq.). The mixture was stirred for 12 hours at 130°C in an atmosphere of CO (g) at 30 atm. The mixture was concentrated to remove the solvent; the residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1/3) as eluent. Methyl 2-(2-((tertbutyldimethylsilyl)oxy)-2-methylpropyl)nicotinate was obtained. LCMS (ES) [M+1] ⁺ m/z: 324.

Stage 4.

Methyl 2-(2-((tert-butyldimethylsilyl)oxy)-2methylpropyl)nicotinate (2.0 g, 6.18 mmol, 1.0 eq.), MeOH (16 ml.) was placed in a 50 ml round bottom flask. and H2O (8 ml). LiOH (520 mg, 12.26 mmol, 2.0 eq.) was then added at 0°C. The mixture was stirred for 2 hours at 50°C. After cooling to room temperature, the pH of the solution was adjusted to 7 using citric acid. The solids were collected by filtration and dried under an infrared lamp. 2-(2((tert-butyldimethylsilyl)oxy)-2-methylpropyl)nicotinic acid was obtained. LCMS (ES) [M+1] ⁺ m/z: 310.

Stage 5.

2-(2-((tert-butyldimethylsilyl)oxy)2-methylpropyl)nicotinic acid (1.60 g, 5.17 mmol, 1.0 eq.), (3R)-morpholine was placed into a 50 mL three-neck round bottom flask. -3-ylmethanol hydrochloride (951 mg, 6.19 mmol, 1.2 eq.), DMF (16 ml) and N,N-diisopropylethylamine (2.0 g, 15.48 mmol, 3.0 eq.). 1-[Bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3oxide-hexafluorophosphate (2.36 g, 6.21 mmol, 1.20 eq) was then added .) at 0°C. The reaction solution was stirred for 2 hours at room temperature. The reaction was then quenched by adding water (30 ml) and extracted with 3x30 ml ethyl acetate. The combined organic phase was washed with brine (30 mlx3) and dried over anhydrous sodium sulfate. The mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1/1) as eluent. (R)-(2-(2-((tertbutyldimethylsilyl)oxy)-2-methylpropyl)pyridin-3-yl)(3-(hydroxymethyl)morpholino)methanone was obtained. LCMS (ES) [M+1] ⁺ m/z: 409.

Stage 6

(R)-(2-(2-((tert-butyldimethylsilyl)oxy)-2methylpropyl)pyridin-3-yl)(3-( hydroxymethyl)morpholino)methanone (1.0 g, 2.45 mmol, 1.0 eq.), 2,6dihydroxybenzaldehyde (406 mg, 2.94 mmol, 1.2 eq.), PPh ₃ (770 mg, 2.0 eq.) 94 mmol, 1.2 eq.) and THF (50 ml). Diisopropyl azodicarboxylate (594 mg, 2.94 mmol, 1.2 eq.) was then added at 0°C. The mixture was stirred overnight at room temperature. The mixture was concentrated to remove the solvent and the residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1/1). (S)-2-((4-(2-(2-((tert-butyldimethylsilyl)oxy)-2methylpropyl)nicotinoyl)morpholin-3-yl)methoxy)-6-hydroxybenzaldehyde was obtained. LCMS (ES) [M+1] ⁺ m/z: 529.

Stage 7.

(S)-2-((4-(2-(2-((tert-butyldimethylsilyl)oxy)-2methylpropyl)nicotinoyl)morpholin-3-yl)methoxy)-6-hydroxybenzaldehyde (600 mg) was placed in a 20 ml vial , 1.14 mmol, 1.0 eq.), CH3CN (5.0 ml) and HCOOH (1.0 ml). The mixture was stirred for 1 hour at 50°C. After cooling to room temperature, the reaction solution was purified directly using preparative-HPLC under conditions: column, Ascentis Express C18, 50 * 3.0 mm, 2.7 μm, mobile phase A:

- 87 045055 water/0.05% formic acid, mobile phase B: CH3CN, flow rate: 1.5 ml/min, gradient: 5% to 100% B over 1.2 minutes, retention 0.6 minutes. (S)-2-hydroxy-6-((4-(2-(2hydroxy-2-methylpropyl)nicotinoyl)morpholin-3-yl)methoxy)benzaldehyde was obtained. LCMS (ES, m/z): [M+H]+: 415.2. ^1H NMR (300 MHz, DMSO^, ppm): δ 11.75 (broad s, 1H), 10.20 (s, 1H), 8.57 (s, 1H), 7.78-7 .28 (m, 3H), 6.81-6.53 (m, 2H), 5.19-4.35 (m, 4H), 4.11-3.88 (m, 1H), 3.71 -3.35 (m, 4H), 3.15-2.59 (m, 3H), 1.31-0.92 (m, 6H).

Example 36. 2-Hydroxy-6-({4-[2-(hydroxymethyl) benzoyl]thiomorpholin-3-yl}methoxy)benzaldehyde, compound 40.

Compound 40 was synthesized according to Scheme 36.

Scheme 36

Stage 1.

A 500 mL three-neck round bottom flask was filled with 2-hydroxymethylbenzoic acid (10.0 g, 65.7 mmol, 1.00 eq), imidazole (8.95 g, 131 mmol, 2.00 eq), and DCM (200 ml.). To this solution was added tert-butyl(chloro)diphenylsilane (21.6 g, 78.8 mmol, 1.20 eq.) dropwise at 0°C. The resulting solution was stirred for 16 hours at 0-25°C. The reaction was then quenched by adding 50 ml of water. The resulting solution was extracted with 3x100 ml DCM. The organic layer was dried and concentrated. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1:50 to 1:1) as eluent. 2-[[(tert-butyldiphenylsilyl)oxy]methyl]benzoic acid was obtained. LCMS (ES) [M+1]+ m/z: 391.2.

Stage 2.

(COCl) ₂ ( 1.30 g, 10.2 mmol, 2.00 eq.) at 0°C. The resulting solution was heated to 40°C for 5 hours. The reaction mixture was then concentrated to obtain a residue. The residue was dissolved in THF (20.0 mL) and triethylamine (1.55 g, 15.3 mmol, 3.00 eq.) was added. To this solution was added thiomorpholin-3-ylmethanol (0.68 g, 5.12 mmol, 1.00 eq.) in portions at 0°C. The resulting solution was stirred for 16 hours at 0-25°C. The reaction was then quenched by adding 10 ml of water. The resulting solution was extracted with 3x20 ml ethyl acetate, dried over anhydrous sodium sulfate and concentrated. The residue was purified on a silica gel column using ethyl acetate/petroleum ether (1:50 to 1:5) as eluent. [4-(2-[[(tertbutyldiphenylsilyl)oxy]methyl]benzoyl)thiomorpholin-3-yl]methanol was obtained. LCMS (ES) [M+1]+ m/z: 506.7.

Stage 3.

[4-(2-[(tert-butyldiphenylsilyl)oxy]methyl]benzoyl)thiomorpholin-3-yl]methanol (1.80 g, 3.55 mmol, 1.00 equiv) was placed in a 100 mL three-neck round bottom flask under _N2 atmosphere. .), 2,6dihydroxybenzaldehyde (0.74 g, 5.33 mmol, 1.50 eq.), PPh ₃ (1.40 g, 5.33 mmol, 1.50 eq.) and DCM (30.0 ml ). To this solution was added a solution of di-tert-butyl azodicarboxylate (1.23 g, 5.33 mmol, 1.5.0 eq.) in DCM (3.0 mL) dropwise at 0°C. The resulting solution was stirred for 15 hours at 025°C. The resulting mixture was concentrated. The residue was applied to a silica gel column with ethyl acetate/petroleum ether (1:100 to 1:1). 2-[[4-(2-[[(tertbutyldiphenylsilyl)oxy]methyl]benzoyl)thiomorpholin-3-yl]methoxy]-6-hydroxybenzaldehyde was obtained. LCMS (ES) [M+1]+ m/z: 626.2.

Stage 4.

2-[[4-(2-[[(tertbutyldiphenylsilyl)oxy]methyl]benzoyl)thiomorpholin-3-yl]methoxy]-6-hydroxybenzaldehyde (1.05 g, 1.67 mmol) was placed in a 100 ml three-neck round bottom flask , 1.00 eq.), THF (5.00 ml) and tetrabutylammonium fluoride (0.33 ml, 0.330 mmol, 0.20 eq.). The resulting solution was stirred for 2 hours at 0-25°C. The resulting mixture was concentrated. The residue was purified on a silica gel column using ethyl acetate/petroleum ether (50:1-1:1) as eluent. 2-hydroxy-6-({4-[2-(hydroxymethyl)benzoyl]thiomorpholin-3yl}methoxy)benzaldehyde was obtained. LCMS (ES) [M+Na]+ m/z: 410.1; 1H NMR (300 MHz, DMSO-d ₆ ) δ 12.00 (s, 1H),

10.38 (s, 1H), 7.51-7.13 (m, 5H), 6.68-6.41 (m, 2H), 5.79-5.48 (m, 1H)), 5 .01-4.30 (m, 6H), 3.84-2.31 (m, 5H).

Example 37 (hydroxymethyl)benzoyl]thiomorpholin-3yl]methoxy}benzaldehyde

Scheme 37

OH OH he

Connection 40

Enantiomer 1 and Enantiomer 2

Compound 40 was purified by chiral-preparative-HPLC (conditions: column: Lux Cellulose-4, 4.6*100 mm, 3 µm; mobile phase, A: n-hexane; B: ethanol (35% B for 18 minutes ); flow rate: 30 ml/min; detector, 220 nm) and analyzed using analytical chiral HPLC (conditions: instrument name: Shimadzu LC-20AD; mobile phase A: n-hexane/DCM = 5/1; mobile phase B : ethanol; column: CHIRALPAK IA-3, 50*4.6 mm, 3 µm IA30CC-UL005). Enantiomer 1 and enantiomer 2 of compound 40 were obtained.

Compound 40, enantiomer 1: analytical chiral HPLC retention time=2.42 min; LCMS (ES) [M+Na]+ m/z: 410.1; 1H NMR (300 MHz, DMSO- _d6 ) δ 11.81-11.70 (m, 1H), 10.32-10.16 (m, 1H), 7.59-7.22 (m, 5H) , 6.76 (d, J= 8.3 Hz, 1H), 6.56 (d, J= 8.4 Hz, 1H), 5.42- 4.41 (m, 6H), 3.47- 3.32 (m, 1H), 3.322.90 (m, 2H), 2.63-2.51 (m, 2H), 2.50-2.40 (m, 1H).

Compound 40, enantiomer 2: analytical chiral HPLC retention time=4.50 min. LCMS (ES) [M+Na]+ m/z: 410.1; 1H NMR (300 MHz, CDCl ₃ ) δ 11.98-11.87 (m, 1H), 10.36 (broad s, 1H), 7.54-7.34 (m, 4H), 7.26- 7.15 (m, 1H), 6.59-6.52 (m, 2H), 5.71-4.35 (m, 5H), 3.91-3.03 (m, 3H), 3.02-2.33 (m, 4H).

Example 38. 2-Hydroxy-6-{[(3K)-4-[2-(hydroxymethyl)benzoyl]thiomorpholin-3-yl]methoxy}benzaldehyde

Scheme 38

OTBDPS

Stage 1.

Phthalide (25.0 g, 186.3 mmol, 1.0 eq), H2O (250 mL) and NaOH (14.91 g, 372.762 mmol, 2 eq) were placed in a 1 L round bottom flask. The resulting solution was stirred for 3 hours at 100°C in an oil bath. The reaction mixture was cooled to 0°C. Solids precipitated after the solution was adjusted to pH 1 with HCl (12 mol/L). The solid product was collected

- 89 045055 filtrate. 2-hydroxymethylbenzoic acid was obtained. LCMS (ES) [M+1]+ m/z: 153.1. 1H NMR (300 MHz, DMSO-d ₆ ) δ 12.88 (broad s, 1H), 7.85 (dd, J= 1.5, 7.8 Hz, 1H), 7.72 (dd, J= 1.8, 7.8 Hz, 1H),

7.57 (td, J= 1.5, 7.5 Hz, 1H), 7.34 (td, J= 1.5, 7.8 Hz, 1H), 4.84 (s, 2H).

Stage 2.

A 500 mL three-neck round bottom flask was filled with 2-hydroxymethylbenzoic acid (15.0 g, 98.6 mmol, 1.0 eq), DCM (200 mL), and imidazole (10.0 g, 147.8 mmol, 1.0 eq.). 5 eq.). After cooling the reaction mixture to 0°C, tert-butyl(chloro)diphenylsilane (32.5 g, 118.3 mmol, 1.2 eq.) was added dropwise with stirring at 0°C. The resulting solution was stirred for 16 hours at 25°C. The reaction was then quenched by adding 300 ml of water. The resulting solution was extracted with 3x250 ml ethyl acetate, dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1:50 to 1:1) as eluent. 2-[[(tert-butyldiphenylsilyl)oxy]methyl]benzoic acid was obtained. LCMS (ES) [M+1] ⁺ m/z: 391.1. ^1H NMR (300 MHz, DMSO-ph) δ 12.88 (s, 1H), 7.93 (td, J= 1.8, 7.8, Hz, 2H), 7.70-7.64 ( m, 5H), 7.50-7.37 (m, 7H), 5.15 (s, 2H), 1.06 (s, 9H).

Stage 3.

2-[[(tert-Butyldiphenyl)oxy]methyl]benzoic acid (15.0 g, 38.4 mmol, 1.0 eq), DCM (250 mL) and two drops of DMF were placed in a 500 mL three-neck round bottom flask. After cooling the reaction mixture to 0°C, (COCl) ₂ (5.8 g, 46.1 mmol, 1.2 eq.) was added dropwise with stirring at 0°C. The resulting solution was stirred for 5 hours at 40°C. The mixture was concentrated under vacuum. 2-[[(tert-butyldiphenylsilyl)oxy]methyl]benzoyl chloride was obtained, which was used in the next step without further purification.

Stage 4.

To a solution of (3R)-thiomorpholin-3-ylmethanol (5.3 g, 40.3 mmol, 1.05 eq.) and triethylamine (7.8 g, 76.7 mmol, 2.0 eq.) in THF ( 250 ml) 2-[[[(tert-butyldiphenylsilyl)oxy]methyl]benzoyl chloride (15.7 g, 38.3 mmol, 1.0 eq.) in THF (50 ml) was added dropwise at 0°C. After addition, the resulting solution was stirred for 5 hours at 0-25°C. The reaction was then quenched by adding 50 ml of water. The resulting solution was extracted with 3x150 ml ethyl acetate, dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1:50 to 1:5) as eluent. [(3R)-4-(2-[[(tert-butyldiphenylsilyl)oxy]methyl]benzoyl)thiomorpholin-3-yl]methanol was obtained. LCMS (ES) [M+1] ⁺ m/z: 506; 1H NMR (300 MHz, DMSO^) δ 7.65-7.63 (m, 5H), 7.62-7.27 (m, 9H), 4.84-4.53 (m, 4H), 3 .80-3.52 (m, 3H), 3.30-2.67 (m, 3H), 2.43-1.99 (m, 2H), 1.06 (s, 9H).

Stage 5.

[(3R)-4-(2-[[(tertbutyldiphenylsilyl)oxy]methyl]benzoyl)thiomorpholin-3-yl]methanol ( 18.0 g, 35.5 mmol, 1.00 eq.), 2,6dihydroxybenzaldehyde (5.4 g, 39.1 mmol, 1.1 eq.), DCM (900.00 ml) and PPh ₃ (14 .0 g, 53.3 mmol, 1.5 eq.). Di-tert-butyl azodicarboxylate (9.8 g, 42.7 mmol, 1.2 eq) in DCM (100 mL) was then added dropwise with stirring at 0°C. The resulting solution was stirred for 15 hours at 0-25°C. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1:100 to 1:1) as eluent. 2-[[(3R)-4-(2-[[(tert-butyldiphenylsilyl)oxy]methyl]benzoyl)thiomorpholin-3-yl]methoxy]-6hydroxybenzaldehyde was obtained. LCMS (ES) [M+1] ⁺ m/z: 626.2; 1H NMR (300 MHz, DMSO- _d6 ) δ 11.71 (s, 1H), 10.19 (s, 1H), 7.61-7.41 (m, 16H), 6.55 (d, J = 8.4 Hz, 1H), 5.23-5.19 (m, 1H), 4.69-4.21 (m, 5H), 3.41-3.37 (m, 2H), 3, 07-2.85 (m, 2H), 2.16-1.99 (m, 1H), 1.06 (s, 9H).

Stage 6.

2-[[(3R)-4-(2-[[(tertbutyldiphenylsilyl)oxy]methyl]benzoyl)thiomorpholin-3-yl]methoxy]-6-hydroxybenzaldehyde (14.0 g, 22.3 mmol, 1.0 eq.) and THF (140 ml). To this solution was added dropwise a solution of tetrabutylammonium fluoride (4.5 ml, 4.50 mmol, 0.20 eq, 1M in THF) at 0°C with stirring. The resulting solution was stirred for 3 hours at 0-25°C. The resulting mixture was concentrated. The residue was applied to a silica gel column with ethyl acetate/petroleum ether (1:50 to 1:1) to obtain the crude product. The crude product was purified by flash preparative-HPLC under the following conditions (IntelFlash-1): column, silica gel C18; mobile phase, MeCN/H ₂ O=1:9, increasing to MeCN/H ₂ O=1:1 within 15; detector, 220. 2-Hydroxy-6-{[(3R)-4-[2(hydroxymethyl)benzoyl]thiomorpholin-3-yl]methoxy} benzaldehyde was obtained, which was subjected to analytical chiral HPLC under the following conditions: Instrument name: Shimadzu LC -20AD; mobile phase A: ngexane/DCM=5/1; mobile phase B: ethanol; column: CHIRALPAK IA-3, 50*4.6 mm, 3 µm IA30CCUL005. Analytical chiral HPLC retention time: 4.540 min. LCMS (ES, m/z): [M+Na]+: 410.1; 1H NMR (300 MHz, DMSO- _d6 ) δ 11.81-11.70 (m, 1H), 10.32-10.10 (m, 1H), 7.59-7.22 (m, 5H) , 6.77-6.55 (m, 2H), 5.42-4.08 (m, 6H), 3.42-3.37 (m, 1H), 3.21-2.90 (m, 2H), 2.71-2.95 (m, 2H), 2.44-2.40 (m,

- 90 045055

1H).

Based on the product of Scheme 38, compound 40, enantiomer 2, was determined to correspond to 2hydroxy-6-{[(3R)-4-[2-(hydroxymethyl)benzoyl]thiomorpholin-3-yl]methoxy}benzaldehyde.

Example 39 {2-[(1S)-1,2-dihydroxyethyl]benzoyl}piperidin-2-yl]methoxy}-6hydroxybenzaldehyde

Scheme 39

Ag ^sn °

39g 'Ό Η

Connection 39h

Diastereomer! and Diastereomer 2

Stage 9a or

Stage 9b

Connection 34

Diastereomer! and Diastereomer 2

Stage 1.

A 500 mL three-neck round bottom flask was filled with 2-bromobenzaldehyde (10.0 g, 54.05 mmol, 1.0 equiv.), methyltriphenyl-lambda-5-hydrobromide (23.20 g, 64.58 mmol, 1.2 eq.) and DMF (100 ml). Four portions of NaH (60% in mineral oil) (9.67 g, 241.69 mmol, 4.5 eq.) were then carefully added at 0°C. The mixture was stirred overnight at room temperature. The reaction was then quenched by adding water/ice (100 ml), extracted with 3x100 ml ethyl acetate. The combined organic phase was washed with brine (80 mlx3) and dried over anhydrous sodium sulfate. The mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using petroleum ether (100%) as eluent. 1-bromo-2-vinylbenzene was obtained. GHMS: 182.

Stage 2.

A 250 mL round bottom flask was filled with 1-bromo-2-vinylbenzene (8.50 g, 46.43 mmol, 1.0 eq.), acetone (130 mL), H ₂ O (13 mL), N-methylmorpholine- N-oxide (5.43 g, 46.35 mmol, 1.0 eq.) and K ₂ OsO ₄ -2H ₂ O (730 mg, 2.32 mmol, 0.05 eq.). The mixture was stirred overnight at room temperature. The mixture was concentrated to remove the solvent and the residue was purified by silica gel column chromatography using THF/petroleum ether (15%) as eluent. 1(2-bromophenyl)ethane-1,2-diol was obtained. LCMS (ES) [M+1]+ m/z: 217.

Stage 3.

1-(2-bromophenyl)ethane-1,2-diol (5.10 g, 23.50 mmol, 1.0 eq.), 2,2-dimethoxypropane (4.17 g, 40.04 mmol, 1.7 eq.), p-toluenesulfonic acid (812 mg, 4.72 mmol, 0.20 eq.) and DMF (75 ml). The reaction solution was stirred for 5 hours at room temperature. The reaction was then quenched by adding water (100 ml) and extracted

- 91 045055

3x100 ml ethyl acetate. The combined organic phase was washed with brine (100 ml*3) and dried over anhydrous sodium sulfate. The mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using petroleum ether (100%) as eluent. 4-(2-bromophenyl)-2,2-dimethyl-1,3-dioxolane was obtained.

GHMS: 256.

Stage 4.

4-(2-bromophenyl)-2,2-dimethyl-1,3-dioxolane (2.50 g, 9.72 mmol, 1 .0 eq.), THF (50 ml). n-BuLi (2.5 M in THF) (4.68 mL, 11.68 mmol, 1.2 eq.) was then added at -78°C. The reaction solution was stirred for 30 minutes at -78°C. Ethyl chloroformate (2.11 g, 19.44 mmol, 2.0 eq) was added to the solution at -78°C. The resulting solution was stirred for an additional 1 hour at room temperature. The reaction was then quenched by adding NH ₄ Cl (aq) (60 ml) and extracted with 3x50 ml ethyl acetate. The combined organic phase was dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1:10) as eluent. Ethyl 2-(2,2-dimethyl-1,3-dioxolan-4yl)benzoate was obtained. GCMS: 250.

Stage 5.

Ethyl 2-(2,2-dimethyl-1,3-dioxolan-4-yl) benzoate (1.60 g, 6.39 mmol, 1.0 eq.), EtOH (10 .0 ml), H ₂ O (50.0 ml) and LiOHH ₂ O (538 mg, 12.82 mmol, 2.0 eq.). The reaction solution was stirred overnight at room temperature. The mixture was concentrated to remove the solvent and the pH of the residue was adjusted to pH 4 with 2 N HCl. The solid was collected by filtration and dried under an infrared lamp. 2-(2,2-dimethyl-1,3dioxolan-4-yl)benzoic acid was obtained. LCMS (ES) [M-1] ⁺ m/z: 221.

Stage 6.

2-(2,2-dimethyl-1,3-dioxolan-4yl)benzoic acid (865 mg, 3.89 mmol, 1.0 eq.), (2S)-piperidine-2 was placed in a 50 mL three-neck round bottom flask. -ylmethanol (537 mg, 4.66 mmol, 1.2 eq.), DMF (20 ml) and N,N-diisopropylethylamine (1.0 g, 7.74 mmol, 2.0 eq.). 1-[Bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide-hexafluorophosphate (1.78 g, 4.68 mmol, 1. 2 equiv.) at 0°C. The mixture was stirred for 2 hours at room temperature. The reaction was then quenched by adding water (30 ml) and extracted with 3x20 ml ethyl acetate. The combined organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated in vacuo. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1/3) as eluent. (2-(2,2-dimethyl-1,3-dioxolan-4yl)phenyl)((S)-2-(hydroxymethyl)piperidin-1-yl)methanone was obtained. LCMS (ES) [M+1] ⁺ m/z: 320.

Stage 7 and stage 8.

(2-(2,2-dimethyl-1,3-dioxolan-4-yl)phenyl)((S)-2(hydroxymethyl) piperidin-1-yl)methanone (900 mg, 2.82 mmol, 1.0 eq.), 2,6-dihydroxybenzaldehyde (467 mg, 3.38 mmol, 1.2 eq.), PPh ₃ (887 mg, 3.38 mmol, 1.20 eq.) and THF (60 ml). Diisopropyl azodicarboxylate (684 mg, 3.38 mmol, 1.2 eq.) was then added at 0°C. After addition, the reaction solution was stirred overnight at room temperature. The mixture was concentrated to remove the solvent and the residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1/1) as eluent.

The collected racemate was separated by chiral-preparative-HPLC under the following conditions: mobile phase: A: n-hexane, B: ethanol, flow rate: 20 ml/min, column: CHIRALPAK IC-3, 4.6x50. mm, 3 µm, gradient: 30% B for 18 minutes, 220 nm.

The separated enantiomers were subjected to analytical chiral HPLC (instrument name: Shimadzu LC-20AD; mobile phase A: n-hexane; mobile phase B: ethanol; column: CHIRALPAK IC-3, 50x4.6 mm, 3 μm IC30CC-SC002). Compound 39h, diastereomer 1 (analytical chiral HPLC conditions: Rt=2.03 min) and compound 39h, diastereomer 2 (analytical chiral HPLC conditions: Rt=2.89 min) were obtained. LCMS (ES) [M+1] ⁺ m/z: 440.

Stage 9a.

Compound 39h, diastereomer 1 (288 mg, 0.66 mmol, 1.0 equiv.), CH3CN (8.0 ml) and Yb(OTf) ₃ (203 mg, 0.33 mmol, 0) were placed in a 25 ml vial. .50 eq.). The mixture was stirred for 2 hours at room temperature. The reaction solution was purified directly by preparative HPLC under the following conditions (SHIMADZU (HPLC-01): column, XBridge C18 OBD Prep Column, 10 µm, 19 mm x 250 mm, mobile phase, water (0.1% formic acid) and CH3CN ( from 5% to 50% phase B for 12 minutes), detector, UV 254 nm. Compound 34 was obtained, diastereomer 1. LCMS (ES, m/z): [M+H] ⁺ : 400. 1H-NMR (300 MHz, DMSO-d ₆ , ppm: δ 11.83-11.73 (m, 1H), 10.30-10.19 (m, 1H), 7.57-7.21 (m, 5H), 6.79-6.53 (m, 2H), 5.25-4.56 (m, 6H), 3.47-2.88 (m, 4H), 1.93-1.37 ( m, 6H).

- 92 045055

Stage 9b.

Compound 39h, diastereomer 2 (307 mg, 0.70 mmol, 1.0 eq.), CH3CN (8.0 ml), Yb(OTf) ₃ (203 mg, 0.35 mmol, 0.50 eq.). The reaction solution was stirred for 2 hours at room temperature. The reaction solution was purified directly by preparative-HPLC under the following conditions (SHIMADZU (HPLC-01): column, XBridge C18 OBD Prep Column, 10 μm, 19 mm x 250 mm, mobile phase, water (0.1% formic acid) and CH3CN (5% to 50% phase B over 12 minutes), detector, UV 254 nm. Compound 34 is obtained, diastereomer 2. LCMS (ES, m/z): [M+H]+: 400. ¹ H NMR (300 MHz, DMSO-66, ppm): δ 11.83-11.73 (m, 1H), 10.33-10.10 (m, 1H), 7.59-7.20 (m , 5H), 6.79-6.53 (m, 2H), 5.25-4.56 (m, 6H), 3.47-2.88 (m, 4H), 1.94-1.37 (m, 6H).

Example 40 -6- {[(3 S)-4- {2-[(1 R)-1 -hydroxyethyl]pyridine-3 carbonyl}morpholin-3-yl]methoxy}benzaldehyde

Compound 41, diastereomer 1 and compound 41, diastereomer 2 were synthesized according to Scheme 40.

Scheme 40

Connection 40 g

Diastereomer! and Diastereomer2

Compound 41 Diastereomer! and Diastereomer2

Stage 1.

A mixture of 3-bromopyridine-2-carbaldehyde (10.0 g, 53.7 mmol, 1.00 eq) and tetrahydrofuran (150 ml) was placed in a 250 mL three-neck round bottom flask, purged and maintained under an inert nitrogen atmosphere. and bromo(methyl)magnesium (35.8 mL, 2.0 eq.) was added dropwise at -78°C. The solution was stirred for 30 minutes at -78°C and then allowed to warm to room temperature for 30 minutes. The reaction was then quenched by adding 100 ml. The resulting solution was extracted with 3x100 ml of ethyl acetate. The resulting mixture was washed with 1x100 ml of saline solution. The mixture was dried over anhydrous sodium sulfate and concentrated. 1-(3-bromopyridin-2-yl)ethanol was obtained. LCMS (ES) [M+1]+ m/z: 202.

Stage 2.

A mixture of 1-(3-bromopyridin-2-yl)ethanol (8.00 g, 39.5 mmol, 1.00 eq.), DMF (80.0 mL), tert-butyl( chloro)diphenylsilane (16.3 g, 59.3 mmol, 1.50 eq.) and imidazole (5.39 g, 79.1 mmol, 2.00 eq.). The resulting solution was stirred for 16 hours at room temperature. The reaction was then quenched by adding 500 ml of water. The resulting solution was extracted with 3x150 ml of ethyl acetate. The resulting mixture was washed with 1x150 ml saline solution. The mixture was dried over anhydrous sodium sulfate and concentrated. The residue was applied to a silica gel column with ethyl acetate/petroleum ether (1/9). 3-bromo-2-[1-[(tertbutyldiphenylsilyl)oxy]ethyl]pyridine was obtained. LCMS (ES) [M+1]+ m/z: 440.1.

Stage 3.

A mixture of 3-bromo-2-[1-[(tert-butyldiphenylsilyl)oxy]ethyl]pyridine (14.0 g, 31.7 mmol, 1.00 eq.), methanol (200 ml) was placed in a 1000 ml high-pressure reactor. , Et ₃ N (6.43 g, 63.5 mmol, 2.00 eq.) and Pd(dppf)Cl ₂ (2.33 g, 3.18 mmol, 0.10 eq.). The air was pumped out of the reactor and the reactor was purged three times with nitrogen, followed by purging with 30 atm CO. The resulting solution was stirred for 16 hours at 110°C. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (2/23) as eluent. Methyl 2-[1-[(tert-butyldiphenylsilyl)oxy]ethyl]pyridine-3-carboxylate was obtained. LCMS (ES) [M+1] ⁺ m/z: 420.2.

Stage 4.

A mixture of methyl 2-[1-[(tert-butyldiphenylsilyl)oxy]ethyl]pyridine-3-carboxylate (10.0 g, 23.8 mmol, 1.00 eq.), MeOH (100 mL) was placed in a 250 mL round bottom flask ) and LiOH (1.71 g, 71.4 mmol, 3.00 eq). The resulting solution was stirred for 3 hours at 50°C. The resulting mixture was concentrated. The resulting solution was diluted with 100 ml of H ₂ O. The pH of the solution was adjusted to 3 using HCl (2 mol/l). Solids were collected by filtration. 2-[1[(tert-butyldiphenylsilyl)oxy]ethyl]pyridine-3-carboxylic acid was obtained. LCMS (ES) [M+1] ⁺ m/z: 406.2.

Stage 5.

A mixture of 2-[1-[(tert-butyldiphenylsilyl)oxy]ethyl]pyridine-3-carboxylic acid (7.00 g, 17.6 mmol, 1.00 eq.), DCM (100 mL) was placed in a 250 mL round bottom flask. , (3R)-morpholin-3-ylmethanol hydrochloride (3.45 g, 22.4 mmol, 1.30 eq.), N,Ndiisopropylethylamine (6.69 g, 51.7 mmol, 3.0 eq.) and 1-[bis-(dimethylamino)methylene]-1H-1,2,3triazolo[4,5-b]pyridinium-3-oxide-hexafluorophosphate (7.88 g, 20.7 mmol, 1.2 eq.)) . The resulting solution was stirred for 3 hours at room temperature. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (3/2) as eluent. [(3R)-4-(2-[1-[(tert-butyldiphenylsilyl)oxy]ethyl]pyridin-3-carbonyl)morpholin-3-yl]methanol was obtained. LCMS (ES) [M+1] ⁺ m/z: 505.3.

Stage 6.

A mixture of [(3R)-4-[2-[(1S)-1-[(tert-butyldiphenylsilyl)oxy]ethyl]pyridine-3-carbonyl was placed into a 250 ml three-neck round-bottom flask, which was purged and maintained inside an inert nitrogen atmosphere. ]morpholin-3-yl]methanol (2.0 g, 3.96 mmol, 1.00 eq.), DCM (100 ml), 2,6-dihydroxybenzaldehyde (0.71 g, 5.15 mmol, 1. 30 eq.) and PPh ₃ (1.56 g, 5.94 mmol, 1.50 eq.). Di-tert-butyl azodicarboxylate (1.00 g, 4.35 mmol, 1.10 eq.) was added dropwise at 0°C. The resulting solution was stirred for 16 hours at room temperature. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1/1) as eluent.

Compound 40g, diastereomer 1 (LCMS retention time: 1.896 min) and compound 40g, diastereomer 2 (LCMS retention time: 1.872 min, (ES) [M+1] ⁺ m/z: 625.2) were obtained. LCMS analysis conditions: instrument: Shimadzu LC2020; column: Kinetex ХВ-С18, 50x3.0 mm, particle size 2.6 µm; mobile phase A: water/0.05% trifluoroacetic acid; mobile phase B: acetonitrile/0.05% trifluoroacetic acid; gradient: 5-100% B for 3 minutes.

Stage 7A.

A solution of Compound 40g, diastereomer 1 (400 mg, 0.640 mmol, 1.00 eq), THF (4.00 ml) and tetrabutylammonium fluoride/THF (3.21 ml, 3.20 mmol, 5) was placed in a 20 ml vial. .00 eq.). The resulting solution was stirred for 3 hours at 45°C. The residue was applied to a silica gel column with ethyl acetate/petroleum ether (99/1~/9). The crude reaction mixture was filtered and subjected to preparative reverse phase HPLC (CV-C18, 50-250 mm, 10 mM; gradient elution from 10% MeCN in water to 45% MeCN in water over a 20-minute period, where both solvents contained 0 .1% formic acid), and the product was analyzed using analytical chiral HPLC (instrument: Shimadzu LC-20AD; mobile phase A: n-hexane (0.1% trifluoroacetic acid); mobile phase B: EtOH/MeOH = 1/1 ; phase B concentration: 20.0%; column: CHIRALPAK IC-3, 50*4.6 mm, 3 μm IC30CC-SC002; column ID: AY30CC-SK001; flow rate: 1,000 ml/min). Compound 41, diastereomer 1, was obtained. Analytical chiral HPLC retention time = 5.801 min. LCMS (ES) [M+1] ⁺ m/z: 387.1. ^1H NMR (300 MHz, DMSO-di) δ 11.84-11.69 (m, 1H), 10.35-10.14 (m, 1H), 8.58-8.54 (m, 1H) , 7.69-7.32 (m, 3H), 6.75-6.54 (m, 2H), 5.33-4.21 (m, 5H), 4.20-3.63 (m, 4H), 3.60-3.35 (m, 1H), 3.23-2.91 (m, 1H), 1.51-1.25 (m, 3H).

Stage 7B.

- 94 045055

A solution of Compound 40g, diastereomer 2 (500 mg, 0.800 mmol,

1.00 equiv.), THF (5.00 ml) and tetrabutylammonium fluoride (4.01 ml, 4.00 mmol, 5.00 equiv.). The resulting solution was stirred for 3 hours at 45°C. The residue was applied to a silica gel column with ethyl acetate/petroleum ether (99/1~1/9). The crude reaction mixture was filtered and subjected to reverse phase preparative HPLC (XB-C18, 50-250 mm, 10 mM; gradient elution from 10% MeCN in water to 45% MeCN in water over a 20-minute period when both solvents contained 0 .1% formic acid), and the product was analyzed using analytical chiral HPLC (instrument: Shimadzu LC-20AD; mobile phase A: n-hexane (0.1% trifluoroacetic acid); mobile phase B: EtOH/MeOH = 1/1 ; phase B concentration: 20.0%; column: CHIRALPAK IC-3, 50*4.6 mm, 3 μm IC30CCSC002; column ID: AY30CC-SK001; flow rate: 1,000 ml/min). Compound 41, diastereomer 2, was obtained. Analytical chiral HPLC retention time = 4.128 min. LCMS (ES) [M+1]+ m/z: 387.1. 1H NMR (300 MHz, DMSO^) δ 11.84-11.20 (broad s, 1H), 10.38-10.15 (m, 1H), 8.588.53 (m, 1H), 7.70- 7.30 (m, 3H), 6.77-6.51 (m, 2H), 5.33-4.75 (m, 3H), 4.55-3.63 (m, 7H), 3. 21-3.02 (m, 1H), 1.51-1.10 (m, 3H).

Example 41. 2-G hydroxy-6-{[(3 S)-4-{2-[(2S)-2-hydroxypropyl]pyridin-3-carbonyl}morpholin-3 yl]methoxy}bechaldehyde and 2-hydroxy- 6-{[(3S)-4-{2-[(2R)-2-hydroxypropyl]pyridine-3carbonyl}morpholin-3-yl]methoxy}benzaldehyde.

Compound 42, diastereomer 1 and compound 42, diastereomer 2 were synthesized according to Scheme 41.

Scheme 41

Connection 42

Diastereomer! and Diastereomer 2

Stage 1.

3-Bromo-2-methylpyridine (25 g, 145.33 mmol, 1.00 eq.) and THF (500.00 mL) were placed in a 1000 mL three-neck round bottom flask, purged and maintained under an inert nitrogen atmosphere. Lithium diisopropylamide (87.20 mL, 174.40 mmol, 1.20 eq) was then added dropwise with stirring at -78°C.

The resulting solution was stirred for 1 hour at -78°C. Acetaldehyde (7.04 g, 159.81 mmol, 1.10 eq) was added dropwise to this mixture with stirring at -78°C. The resulting solution was stirred for 1 hour at -78°C. The reaction was then quenched by adding 300 ml of saturated NH ₄ Cl solution. The resulting mixture was extracted with 3x300 ml ethyl acetate, the organic layers were combined, dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography using THF/petroleum ether (20%) as eluent to give 1-(3-bromopyridin-2-yl)propan-2-ol. LCMS (ES) [M+1]+ m/z: 216.

Stage 2.

1-(3-bromopyridin-2-yl)propan-2-ol (15.00 g, 69.42 mmol, 1.00 eq.), imidazole (9.45 g, 138, 81 mmol, 2.00 eq.), DMF (300.00 ml), 4dimethylaminopyridine (0.85 g, 6.94 mmol, 0.1 eq.) and tert-butyl(chloro)diphenylsilane (22.90 g, 83.30 mmol, 1.20 eq.). The resulting solution was stirred overnight at 60°C. The reaction mixture was cooled to room temperature. The reaction was then quenched by adding 300 ml of water. The resulting solution was extracted with 3x300 ml ethyl acetate, the organic layers were combined, dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography using THF/petroleum ether (5%) to give 3-bromo-2-[2-[(t-butyldiphenylsilyl)oxy]propyl]pyridine. LCMS (ES) [M+1] ⁺ m/z: 454.

Stage 3.

3-bromo-2-[2-[(tert-butyldiphenylsilyl)oxy]propyl]pyridine (25.00 g, 55.00 mmol, 1.00 eq.), MeOH (800.00 ml) was placed in a 2000 ml pressure reactor ), triethylamine (11.13 g, 110.01 mmol, 2.00 eq.) and Pd(dppf)Cl ₂ (4.02 g, 5.49 mmol, 0.10 eq.). The air was pumped out of the reactor and the reactor was purged three times with nitrogen, followed by purging with 30 atm CO. The resulting solution was stirred overnight at 110°C. The reaction mixture was cooled to room temperature. The resulting mixture was concentrated. The residue was purified by silica gel column chromatography using THF/petroleum ether (7%) to give methyl 2-[2-[(t-butyldiphenylsilyl)oxy]propyl]pyridine-3-carboxylate. LCMS (ES) [M+1] ⁺ m/z: 434.

Stage 4.

Methyl 2-[2-[(tert-butyldiphenylsilyl)oxy]propyl]pyridine-3-carboxylate (20.00 g, 46.12 mmol, 1.00 eq.), MeOH (400 mL) was placed in a 1000 mL round bottom flask. , H ₂ O (200 ml) and LiOH-H ₂ O (3.87 g, 92.22 mmol, 2.00 eq.). The resulting solution was stirred for 4 hours at 50°C. The reaction mixture was cooled to room temperature. The resulting mixture was concentrated. The resulting solution was extracted with 200 ml of ethyl acetate and the aqueous layers were combined. The pH value of the solution was adjusted to 4-5 using HCl (1 mol/l). The resulting precipitates were collected by filtration and dried under infrared light. 2-[2-[(tert-butyldiphenylsilyl)oxy]propyl]pyridine-3-carboxylic acid was obtained. LCMS (ES) [M+1] ⁺ m/z: 420.

Stage 5.

2-[2-[(tert-Butyldiphenylsilyl)oxy]propyl]pyridine-3-carboxylic acid (4.00 g, 9.53 mmol, 1.00 eq.), (3R)morpholine- 3-ylmethanol hydrochloride (1.76 g, 11.46 mmol, 1.20 eq), DCM (100.00 ml) and N,Ndiisopropylethylamine (3.70 g, 28.59 mmol, 3.00 eq) . After this, 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide-hexafluorophosphate (4.35 g, 11.44 mmol, 1. 20 equiv.) at 0°C. The resulting solution was stirred for 3 hours at room temperature. The reaction was then quenched by adding 100 ml of water. The resulting solution was extracted with 3x100 ml dichloromethane, the organic layers were combined, dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography using THF/petroleum ether (25%) as eluent to give [(3R)-4-(2-[2-[(tert-butyldiphenyl)oxy]propyl]pyridine-3-carbonyl )morpholin-3-yl]methanol. LCMS (ES) [M+1] ⁺ m/z: 519.

Stage 6.

2,6-Dihydroxybenzaldehyde (0.96 g, 6.94 mmol, 1.20 equiv), [(3R)-4-(2-[2-[(tert-butyldiphenylsilyl)) was placed in a 250 mL three-neck round bottom flask. oxy]propyl]pyridin-3carbonyl)morpholin-3-yl]methanol (3.00 g, 5.78 mmol, 1.00 eq.), PPh ₃ (1.82 g, 6.94 mmol, 1.20 eq. .) and DXM (100.00 ml). Diisopropyl azodicarboxylate (1.40 g, 6.92 mmol, 1.20 eq) was then added dropwise with stirring at 0°C. The resulting solution was stirred overnight at room temperature. The resulting mixture was concentrated. The residue was purified by a silica gel column using THF/petroleum ether (30%) as eluent to obtain the crude product. The crude product was purified by preparative-HPLC under the following conditions: column: Welch XBC18 50x250 mm, 10 μm, mobile phase, water (0.1% trifluoroacetic acid) and acetonitrile (50% for 100 minutes); detector, 254. Compound 41g, diastereomer 1 (retention time=70 minutes) and compound 41g, diastereomer 2 (retention time=90 minutes) were obtained. LCMS (ES) [M+1] ⁺ m/z: 639.

Stage 7A.

Compound 41g, diastereomer 1 (1.2 g, 1.88 mmol, 1.00 eq), THF (9.00 mL) and tetrabutylammonium fluoride/THF (9.39 mL, 9.39 mmol, 5.00 eq.). The resulting solution was stirred for 5 hours at 45°C. The reaction mixture was cooled to room temperature. The crude product was purified by preparative-HPLC under the following conditions: column, XBridge Prep C18 OBD column, 19 cm, 150 mm, 5 µm; mobile phase, water (0.1% HCOOH) and acetonitrile (5% to 55% for 15 minutes); detector, 254. Compound 42 was obtained, diastereomer 1. Final

- 96 045055 This compound was analyzed using chiral HPLC under the following conditions: instrument: SHIMADZU LC-20AT; mobile phase A: n-hexane; mobile phase B: mobile phase B; conc. phase B: 50.0%, column flow rate: 1,000 ml/min: CHIRALPAK AY-3, 4.6*50 mm, 3 µm; Column ID: AY30CC-SK001; retention time=3.35 min. LCMS: (ES, m/z): [M+H]+: 401. 1H NMR (300 MHz, DMSO-d6, ppm): δ 11.80-11.67 (m, 1H), 10 ,34-10.23 (m, 1H), 8.58 (dd, J= 4.9, 1.7 Hz, 1H), 7.75 (d, J= 7.9 Hz, 1H), 7, 57 (t, J= 8.4 Hz, 1H), 7.33 (d, J= 5.7 Hz, 1H), 6.75 (d, J= 8.1 Hz, 1H), 6.55 ( dd, J= 8.5, 4.9 Hz, 1H), 5.04-4.89 (m, 1H), 4.49-4.29 (m, 4H), 4.09 (d, J= 12.1 Hz, 1H), 3.99-3.63 (m, 3H), 3.57-3.07 (m, 3H), 2.94-2.60 (m, 1H), 1.19 -0.81 (m, 3H).

Stage 7B.

Compound 41g, diastereomer 2 (1.20 g, 1.88 mmol, 1.00 eq), THF (9.00 mL) and tetrabutylammonium fluoride (9.39 mL, 9.39 mmol, 5.00 eq.). The resulting solution was stirred for 5 hours at 45°C. The reaction mixture was cooled to room temperature. The crude product was purified by preparative-HPLC under the following conditions: column, XBridge Prep C18 OBD column, 19 cm, 150 mm, 5 µm; mobile phase, water (0.1% HCOOH) and acetonitrile (5% to 55% for 15 minutes); detector, 254. Compound 42, diastereomer 2 was obtained. The final compound was analyzed by chiral HPLC under the following conditions: instrument: SHIMADZU LC-20AT; mobile phase A: n-hexane; mobile phase B: mobile phase B; conc. phase B: 50.0%; column flow rate: 1,000 ml/min: CHIRALPAK AY-3, 4.6*50 mm, 3 µm; Column ID: AY30CC-SK001; retention time=1.91 min. LCMS: (ES, m/z): [M+H]+: 401. 1H NMR: (300 MHz, DMSO-b6, ppm): δ 11.82-11.69 (m, 1H), 10.33-10.23 (m, 1H), 8.58 (dd, J= 4.9, 1.8 Hz, 1H), 7.84-7.26 (m, 3H), 6.76 ( d, J= 8.3 Hz, 1H), 6.56 (d, J= 8.4 Hz, 1H), 4.95 (s, 1H), 4.45-4.15 (m, 4H), 4.09-3.61 (m, 4H), 3.46-3.12 (m, 3H), 2.95-2.66 (m, 1H), 1.20-0.83 (m, 3H )).

Example 42 [(3R)-4-2-[( 1R)-1 -hydroxyethyl]pyridine-3carbonyl}thiomorpholin-3-yl]methoxy}benzaldehyde.

Compound 43, diastereomer 1 and compound 43, diastereomer 2 were synthesized according to Scheme 42.

Scheme 42

Connection 43

Diastereomer! and Diastereomer?

Stage 1.

To a solution of 2-[1-[(tert-butyldiphenylsilyl)oxy]ethyl]pyridine-3-carboxylic acid (2.0 g, 4.93 mmol, 1.0 eq.)) in DMF (20.0 ml) was added diisopropylethylamine (1.27 g, 9.8 mmol, 2.0 eq.) and 1[bis-(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide -hexafluorophosphate (2.25 g, 5.9 mmol, 1.2 eq.) at 0°C. After stirring the reaction mixture at 0°C for 20 minutes, (3R)-thiomorpholin-3-ylmethanol (720 mg, 5.42 mmol, 1.10 eq.) was added in portions. The resulting solution was stirred for 2 hours at 25°C. The reaction was then quenched by adding 50 ml of water. The resulting solution was extracted with 3x80 ml ethyl acetate. The combined organic layer was dried over anhydrous sodium sulfate and concentrated in vacuo. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1:100 to 1:1) as eluent. [(3R)-4-(2-[1-[(tert-butyldiphenylsilyl)oxy]ethyl]pyridine-3-carbonyl)thiomorpholin-3yl]methanol was obtained. LCMS (ES) [M+1]+ m/z: 521.2; ¹ H NMR: (300 MHz, DMSO-d ₆ , ppm): δ 8.75 (d, J=5.4 Hz, 1H), 7.96 (s, 1H), 7.71-7 .26 (m, 11H), 4.98-4.44 (m, 3H), 4.07-3.54 (m, 2H), 3.12-2.97 (m, 1H), 2.91 -2.84 (m, 1H), 2.74-2.70 (m, 1H), 2.38-2.33 (m, 1H), 1.79-1.69 (m, 1H), 1 .59-1.10 (m, 3H), 0.92 (s, 9H).

Stage 2.

[(3R)-4-(2-[1-[(tert-butyldiphenylsilyl)oxy]ethyl]pyridine-3-carbonyl)thiomorpholin-3yl]methanol solution (1.3 g, 2.5 mmol, 1.0 equiv .), 2,6-dihydroxybenzaldehyde (380 mg, 2.72 mmol, 1.1 eq.) and PPh3 (980 mg, 3.75 mmol, 1.5 eq.) in DCM (200 ml) were cooled to 0° C in an argon atmosphere. Drop by drop until

- 97 045055 a solution of di-tert-butyl azodicarboxylate (690 mg, 3.0 mmol, 1.2 eq.) in DCM (30 ml) was added. After addition, the resulting solution was stirred for 16 hours at 0-25°C. The reaction was concentrated under vacuum. The residue was purified on a silica gel column using ethyl acetate/petroleum ether (1:100 to 1:5) as eluent. 2-[[(3R)-4-(2-[1-[(tertbutyldiphenylsilyl)oxy]ethyl]pyridin-3-carbonyl)thiomorpholin-3-yl]methoxy]-6-hydroxybenzaldehyde was obtained. LCMS (ES) [M+1]+ m/z: 641.2; 1H NMR (300 MHz, _CDCl3 , ppm): δ 11.96 (s, 1H), 10.25 (broad s, 1H), 8.89-8.78 (m, 1H), 7. 81-7.19 (m, 13H), 6.63-6.28 (m, 2H), 5.21-4.89 (m, 2H), 4.45-4.13 (m, 2H), 3.71-3.66 (m, 1H), 3.18-2.92 (m, 2H), 2.75-2.35 (m, 3H), 1.74 -1.50 (m, 3H ), 0.92 (s, 9H).

Stage 3.

2-[[(3R)-4-(2-[1-[(tertbutyldiphenylsilyl)oxy]ethyl]pyridin-3-carbonyl)thiomorpholin-3-yl]methoxy]-6-hydroxybenzaldehyde was placed in a 100 ml three-neck round bottom flask (2.0 g, 3.12 mmol, 1.0 eq.) and THF (10 ml). After cooling the reaction mixture to 0° C., tetrabutylammonium fluoride (1.63 g, 6.24 mmol, 2.0 eq.) was added in portions. The resulting solution was stirred for 5 hours at 45°C. The resulting solution was quenched with NH ₄ Cl (20 ml, 2 N) and extracted with ethyl acetate (50 ml x 3); the organic layers were combined and concentrated. The residue was purified on a silica gel column, eluting with petroleum ether/ethyl acetate=100:1 to 1:1, to obtain the racemate, which was purified by preparative chiral-HPLC under the following conditions: column: Lux Amylose-1, 50*250 mm , 10 µm; mobile phase: A: n-hexane; B: ethanol; flow rate: 90 ml/min; gradient: 50% B for 50 minutes; 220 nm. The isolated diastereomers were analyzed using analytical HPLC under the following conditions: instrument: SHIMADZU LC-20AT; mobile phase A: n-hexane; mobile phase B: ethanol; conc. phase B: 50.0%; flow rate: 1,000 ml/min; column: Lux Amylose-1, 4.6*100 mm, 3 µm; Column ID: H18-344853. Compound 43, diastereomer 1 and compound 43, diastereomer 2 were obtained.

Data for compound 43, diastereomer 1: chiral HPLC retention time 8.31 min; LCMS (ES, m/z): [M+H]+: 403.1; 1H NMR: (300 MHz, DMSO, ppm): δ 11.85 (broad s, 1H), 10.3510.16 (m, 1H), 8.57-8.53 (m, 1H), 7.75-7.32 (m, 3H), 6.77-6.55 (m, 2H), 5.42-5.27 (m, 2H), 4.88-4.03 (m, 3H ), 3.47-3.44 (m, 2H), 3.21-2.73 (m, 3H), 2.50-2.44 (m, 1H), 1.43-1.34 (m , 3H).

Data for compound 43, diastereomer 2: chiral HPLC retention time 5.30 min; LCMS (ES, m/z): [M+H]+: 403.1; 1H NMR: (300 MHz, DMSO, ppm): δ 11.79 (broad s, 1H), 10.3210.16 (m, 1H), 8.60-8.53 (m, 1H), 7.77-7.28 (m, 3H), 6.77-6.55 (m, 2H), 5.43-5.5.33 (m, 2H), 4.88-4.06 (m , 3H), 3.50-3.34 (m, 2H), 3.15-2.36 (m, 4H), 1.46-1.34 (m, 3H).

Example 43 [(3R)-4-{2-[(2R)-2-hydroxypropyl]pyridine-3carbonyl}thiomorpholin-3-yl]methoxy}benzaldehyde.

Compound 44, diastereomer 1 and compound 44, diastereomer 2 were synthesized according to Scheme 43.

Scheme 43

Connection 44

Diastereomer! and 2

Stage 1.

Diisopropylethylamine was added to a solution of 2-[2-[(tert-butyldiphenylsilyl)oxy]propyl]pyridine-3-carboxylic acid (2.0 g, 4.76 mmol, 1.0 eq.) in DMF (20.0 mL). (1.23 g, 9.5 mmol, 2 eq.) and 1[bis-(dimethylαamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide-hexafluorophosphate ( 2.17 g, 5.720 mmol, 1.20 eq.) at 0°C. After stirring the reaction mixture at 0°C for 20 minutes, (3R)-thiomorpholin-3-ylmethanol (0.70 g, 5.243 mmol, 1.1 eq.) was added in portions. The resulting solution was stirred for 2 hours at 25°C. The reaction was then quenched by adding 50 ml of water. The resulting solution was extracted with 3x80 ml of ethyl acetate, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1:100 to 1:1) as eluent. [(3R)-4-(2-[2-[(tertbutyldiphenylsilyl)oxy]propyl]pyridin-3-carbonyl)thiomorpholin-3-yl]methanol was obtained. LCMS (ES) [M+1]+ m/z: 535.2.

Stage 2.

[(3R)-4-(2-[2-[(tert-butyldiphenylsilyl)oxy]propyl]pyridine-3-carbonyl)thiomorpholine- 3-yl]methanol (1.5 g, 2.8 mmol, 1.0 eq.), 2,6dihydroxybenzaldehyde (0.43 g, 3.1 mmol, 1.1 eq.), DCM (150.00 ml ) and PPh ₃ (1.1 g, 4.2 mmol, 1.5 eq.). After the reaction mixture had cooled to 0°C, a solution of di-tert-butyl azodicarboxylate (0.78 g, 3.36 mmol, 1.2 eq) in DCM (10 ml) was added dropwise. The resulting solution was stirred for 16 hours at a temperature from 0 to 25°C. The resulting mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography using ethyl acetate/petroleum ether (1:100 to 1:5) as eluent. 1-[[(3R)-4-(2-[2-[(tertbutyldiphenylsilyl)oxy]propyl]pyridine-3-carbonyl)thiomorpholin-3-yl]methoxy]-6-hydroxybenzaldehyde was obtained. LCMS (ES) [M+1] ⁺ m/z: 655.2; ^1H NMR: (300 MHz, DMSO- _d6 , ppm): δ 11.76 (broad s, 1H), 10.39 (broad s, 1H), 8.54-8.48 (m, 1H), 7.86-7.27 (m, 13H), 6.78-6.55 (m, 2H), 5.41 (broad s, 1H), 4.83-4.44 (m, 3H ), 3.39-3.14 (m, 4H), 3.10-2.70 (m, 3H), 2.41-2.11 (m, 1H), 1.02-0.81 (m , 12H).

Stage 3.

2-[[(3R)-4-(2-[2-[(tertbutyldiphenylsilyl)oxy]propyl]pyridin-3-carbonyl)thiomorpholin-3-yl]methoxy]-6-hydroxybenzaldehyde was placed in a 100 ml three-neck round bottom flask (1.0 g, 1.52 mmol, 1.0 eq.) and THF (10 ml). After cooling the reaction mixture to 0°C, a solution of triethylamine trihydrofluoride (1.0 g, 80.9 mmol, 3.0 eq.) was added dropwise. The resulting solution was stirred for 5 hours at 45°C. The pH of the solution was adjusted to 8 using NaHCO ₃ (2 mol/l). The reaction mixture was extracted with ethyl acetate (50 mlx3), the organic layers were combined and concentrated. The crude product was purified by silica gel column chromatography using petroleum ether/ethyl acetate=100:1 to 1:1 to give the racemate.

The racemate was purified using chiral-HPLC (conditions: column: Lux Amylose-1, 50*250 mm, 10 µm; mobile phase: A: n-hexane B: ethanol; flow rate: 90 ml/min; gradient: 50% B for 36 minutes; 220 nm) and analyzed by analytical HPLC (Conditions: instrument: SHIMADZU LC-20AT; mobile phase A: n-hexane; mobile phase B: ethanol; phase B concentration: 50.0%; flow rate: 1,000 ml/min; column: Lux Amylose-1, 4.6*100 mm, 3 µm; column ID: H18-344853). Compound 44, diastereomer 1 and compound 44, diastereomer 2 were obtained.

Data for compound 44, diastereomer 1: chiral HPLC retention time = 4.85 min; LCMS (ES, m/z): [M+H]+: 417.2; 1H NMR: (300 MHz, DMSO^, ppm): δ 11.77 (broad s, 1H), 10.33 (s, 1H), 8.56 (dd, J= 1.8, 4, 8 Hz, 1H), 7.76-7.29 (m, 3H), 6.75-6.55 (m, 2H), 5.43-5.41 (m, 1H), 4.81-4 .13 (m, 4H), 3.49-3.41 (m, 2H), 3.11-2.41 (m, 6H), 1.08-0.92 (m, 3H).

Data for compound 44, diastereomer 2: chiral HPLC retention time 6.94 min; LCMS (ES, m/z): [M+H]+: 417.2; 1H NMR: (300 MHz, DMSO^, ppm): δ 10.33 (s, 1H), 8.57-8.48 (m, 1H), 7.80-7.27 (m, 3H ), 6.75-6.54 (m, 2H), 5.53-41 (m, 1H), 4.56-4.06 (m, 4H), 3.58-3.40 (m, 2H ), 3.15-2.67 (m, 5H), 2.43-2.38 (m, 1H), 1.14-0.89 (m, 3H).

Compounds 6-9 and 18 in table. 3 was synthesized in accordance with the methods specified in the present description, and their appropriate modifications.

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Table 3

Connection Number Structure Mass Spectrometry Data 6 / ¹ x ° \ Р /-у р-^у X \ О ^Z \—у Уо О 384.1 (MH+) 7 O o9 {—\ O 386.2 (MH+) 8 .on ⁰ C o OO 387.1 (MH+) 9 \ Ζ ζ^ \ /=\ / | o Q° ¹ o /=o - ¹ - I 371.1 (MH+) 18 he oiy ⁰ % o 00 LCMS (ES) [M+l] ⁺ m/z 399.2

Biological analyses.

Whole blood analysis.

Oxygen equilibrium curves (OEC) were obtained using a TCS Hemox analyzer (TCS Scientific Company, New Hope, PA, USA) to measure changes in O2 binding capacity for hemoglobin. Whole blood was incubated for 1 hour at 37°C with the indicated compounds at an equimolar ratio of hemoglobin to compound and diluted with TES mixture (2-[[1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl]amino]ethanesulfonic acid )/buffered saline before measurements. For example, for whole blood with a hematocrit [Hct] of 20%, corresponding to a 1 mM hemoglobin solution, a compound concentration of 1 mM was used (eg, for compounds 1-5), and the incubated sample was diluted 50 to 100 times. The concentration of compounds 6-44 (Diastereomers 1 and 2) was varied, but it remained in an equimolar ratio to hemoglobin. The diluted samples were then oxygenated using compressed air in a Hemox analyzer and oxygen equilibrium curves were obtained during deoxygenation as previously described (Guarnone et al, Haematologica, 1995, 80, 426-430). p50 values (the partial pressure of O2 at which hemoglobin is 50% saturated with _O2 ) were obtained using nonlinear regression analysis.

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The percentage change in p50 [Δρ50 (%)] was calculated as follows: Δρ50 (%) = [(p50 of the control compound) - p50 of the compound)/p50 of the control compound] x 100. The data obtained are shown in table. 4. Enantiomer 1 and enantiomer 2 of compound 13 also have a Δρ50 of about 61.0% to about

80.6%.

Table 4

Connection Number Delta-p50 (%) 1 77.3 2 84.4 3 85.8 4 75.5 5 81.0 6 74.5 7 62.7 8 79.8 9 62.2 10 (Enantiomer 1) 65.6 10 (Enantiomer 2) 87.3 AND 79.3 12 76.7 13 (Enantiomer 1) 80.6 13 (Enantiomer 2) 61.0 14 74.56 15 80.5 16 49.55 17 27.09 18 79.67 19 54.67 20 78.32 21 60.21 22 70.32 23 70.92 24 65.51 25 53.36 26 51.72 27 66.58 28 83.35 29 78.79 thirty 74.19 31 74.03 32 60.55 33 49.03 34 79.46 34 (Diastereomer 1) 80.43 34 (Diastereomer 2) 81.59 35 (Diastereomer 1) 81.35 35 (Diastereomer 2) 83.1 36 77.04 37 60.58 38 60.85 39 77.98 40 72.82 40 (Enantiomer 1) 64.31 40 (Enantiomer 2) 83.36 41 (Diastereomer 1) 66.71 41 (Diastereomer 2) 53.94 42 (Diastereomer 1) 76.39 42 (Diastereomer 2) 75.05 43 (Diastereomer 1) 64.50 43 (Diastereomer 2) 64.56 44 (Diastereomer 1) 66.57 44 (Diastereomer 2) 54.34

Cytochrome P450 (CYP) Enzyme Activity Assay (Pregnane X Receptor, PXR): Nuclear pregnane X receptor activator was seeded using stably transfected human hepatoma cell lines (DPX2) in a 96-well plate. 24 hours after plating, cells were treated with selected concentrations of compounds in duplicate wells, and then cells were returned to the incubator for an additional 24 hours. At the end of this incubation period, the number of viable cells per well was determined using Promega's Cell Titer cytotoxicity assay.

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Fluor. Additionally, Promega's ONE-Glo was added to the same wells and reporter gene activity was assessed. The average values of luminescence units for each dose of compound for two parallel experiments were divided by the average value for a solvent control such as DMSO to determine the fold induction. Consistent with the industry standard, a threshold of >2.5 times was used to label compounds that have a risk of inducing cytochrome P450 enzymes in vivo.

The structures of the comparative compounds (compound A, compound B and compound C) are given in table below. 5.

Table 5

Comparative compound A Comparative compound B Comparative compound C Structure 0% ⁰ V o L / ^{1 1} °</° L L YAG ⁰ O >=o

The results for the various compounds disclosed herein and selected comparative compounds are summarized in Table 1. 6.

Table 6

Compound Marker of cytochrome P450 enzyme (pregnaneX receptor) Comparative compound A Y Comparative compound B Y Comparative compound C Y ¹ 1 N ¹ 8 N 12 N Compound 13 (Enantiomer 1) N

¹ When

Claims (28)

Table 7 Compound T1/2 (h) Blood/plasma ratio Comparative compound A 29 75 Comparative compound B 29.8 98 1 58 162 8 69 105 10 (Enantiomer 2) 112 212 AND 55 126 12 58 131 20 65 45 23 62 59 36 56 115 39 52 52 40 (Enantiomer 2) 117 424 13 (Enantiomer 1) 88 230 35 (Diastereomer 1) 102 493 35 (Diastereomer 2) 89 636 Unless otherwise specified, all technical and scientific terms used in this specification have the same meaning as commonly understood by one skilled in the art to which the present invention belongs. The present inventions set forth herein by way of illustration may be suitably practiced in the absence of any element or elements, limitations or limitations not specifically disclosed herein. Thus, for example, the terms consisting, including, containing, etc. should be read as open and non-restrictive. Additionally, the terms and expressions used herein are used as terms of description and not limitation, and the use of such terms and expressions is not intended to exclude any equivalents of the features shown and described, or portions thereof, but it is understood that various modifications are possible in within the scope of the claimed invention. The contents of all publications, patent applications, patents and other references mentioned herein are expressly incorporated by reference in their entirety to the same extent as if the contents of each were incorporated by reference individually. In the event of any conflict, this specification, including definitions, governs. CLAIM 1. Compound of formula I or a pharmaceutically acceptable salt thereof, wherein X represents CH or N; Y represents CH or N; Z represents a bond, CH2, O or S and R ¹ represents monohydroxy-(C ₁ . ₄ alkyl), dihydroxy-(C ₁ . ₄ alkyl), -CH ₂ CH ₂ OCH ₃ , -CH2CH2CN or 2. The compound according to claim 1, wherein X represents CH. 3. A compound according to claim 1, wherein X is N. 4. A compound according to any one of claims 1 to 3, wherein Y represents CH. - 103 045055 5. A compound according to any one of claims 1 to 3, wherein Y is N. 6. A compound according to any one of claims 1 to 5, in which Z represents a bond, CH2 or O. 7. A compound according to any one of claims 1 to 5, wherein Z represents a bond. 8. A compound according to any one of claims 1 to 5, in which Z represents CH2. 9. A compound according to any one of claims 1 to 5, in which Z represents O. 10. A compound according to any one of claims 1 to 5, wherein Z is S. 11. Compound according to claim 1 of formula Ia where X represents CH or N and R ¹ represents monohydroxy-(C ₁ . ₄ alkyl), dihydroxy-(C ₁ . ₄ alkyl), -CH2CH2OCH ₃ , -CH2CH2CN or 12. A compound according to any one of claims 1 to 11, in which R ¹ represents -CH2OH, -CH2CH2OH, -CH2CH2CN or 13. A compound according to any one of claims 1 to 11, in which R ¹ represents -CH2OH or -CH2CH2OH. 14. A compound according to any one of claims 1 to 11, in which R ¹ represents monohydroxy-( _C1.4 alkyl) or dihydroxy-( _C1.4 alkyl). 15. A compound according to any one of claims 1 to 11, in which R ¹ represents hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, 1,2-dihydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl or 2-hydroxy-2-methylpropyl. 16. The connection according to claim 1, where the specified connection is selected from - 104 045055 - 105 045055 - 106 045055 or a pharmaceutically acceptable salt of each. - 107 045055 17. Connection selected from - 108 045055 or a pharmaceutically acceptable salt of each. 18. A compound of formula or a pharmaceutically acceptable salt thereof. 19. Compound formula 20. A pharmaceutical composition containing a compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient. 21. A pharmaceutical composition containing a compound according to claim 19 and a pharmaceutically acceptable excipient. 22. A method of increasing the oxygen affinity of hemoglobin type S in a subject in need thereof, comprising administering to the subject a compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 20. 23. A method of increasing the oxygen affinity of hemoglobin type S in a subject in need thereof, comprising administering to the subject a compound of claim 19 or a pharmaceutical composition of claim 21. 24. A method of treating a hemoglobin-mediated disorder in a subject in need thereof, comprising administering to the subject a compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 20. 25. A method of treating a hemoglobin-mediated disorder in a subject in need thereof, comprising administering to the subject a compound of claim 19 or a pharmaceutical composition of claim 21. 26. The method according to claim 24 or 25, wherein the hemoglobin is sickle hemoglobin. - 109 045055 27. A method of treating sickle cell disease in a subject in need thereof, comprising administering to the subject a compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 20. 28. A method of treating sickle cell disease in a subject in need thereof, comprising administering to the subject a compound of claim 19 or a pharmaceutical composition of claim 21. Eurasian Patent Organization, EAPO Russia, 109012, Moscow, Maly Cherkassky lane, 2 - BUT-